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8 TH INTERNATIONAL SYMPOSIUM ON FRUIT<br />
FLIES OF ECONOMIC IMPORTANCE<br />
VALENCIA, SPAIN<br />
From 26 th September to 1 st October 2010
First edition 2010<br />
© Of the present edition: Editorial Universitat Politècnica de València<br />
www.editorial.upv.es<br />
© All commercial names, brands, or distinctive marks of any kind included in this<br />
piece of work are protected by law.<br />
© Edited by Beatriz Sabater Muñoz, Vicente Navarro Llopis and Alberto Urbaneja<br />
© Cover design by Cristial Vidal Quist and María Juan Blasco<br />
© Print:<br />
ISBN: 978-84-693-4446-0<br />
Depósito Legal:<br />
Ref. editorial: 2367<br />
Any unauthorized copying, distribution, marketing, editing, and in general any other exploitation,<br />
for whatever reason, of this piece of work or any part thereof, is strictly prohibited without the<br />
authors’ expressed and written permission.<br />
Printed in Spain
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CONTENTS<br />
Welcome message ...................................................................................................... 5<br />
8 th ISFFEI Committees .................................................................................................. 6<br />
Organizing Committee .............................................................................................. 6<br />
Scientific Committee................................................................................................ 7<br />
Fruit Fly Steering Committee (as in June 2008) ................................................................ 8<br />
Acknowledgement to Official Organizing & Supporting Institutions ........................................... 9<br />
Acknowledgement to Sponsors & Exhibitors ...................................................................... 10<br />
About ISFFEI ............................................................................................................ 11<br />
General information ................................................................................................... 12<br />
Congress language ............................................................................................... 12<br />
Congress technical organizers ................................................................................. 12<br />
Congress venue................................................................................................... 12<br />
Registration desk & Technical secretariat .................................................................. 13<br />
Internet access & Stations ..................................................................................... 13<br />
Shuttle services & Public transport in Valencia city....................................................... 13<br />
Accompanying persons .......................................................................................... 13<br />
Speakers, audio visual & Technical desk .................................................................... 14<br />
Poster presenters ................................................................................................ 14<br />
Abstracts & CD provided ....................................................................................... 15<br />
Special Events .................................................................................................... 15<br />
Coffe breaks & Lunch ........................................................................................... 15<br />
Exhibition ......................................................................................................... 15<br />
SPONSORS ............................................................................................................... 17<br />
EXHIBITORS ............................................................................................................. 25<br />
Program at glance ..................................................................................................... 33<br />
PLENARY TALKS ........................................................................................................ 37<br />
ORAL PRESENTATIONS ................................................................................................. 43<br />
Session 1 Biology, Ecology & Behaviour...................................................................... 45<br />
Session 2 Morphology & Taxonomy ........................................................................... 57<br />
Session 3 Genetics & Evolution ................................................................................ 63<br />
Session 4 Risk Assessment, Quarantine & Post-harvest Treatments .................................... 73<br />
Session 5 SIT Principles & Application ....................................................................... 79<br />
Session 6 Area-Wide & Action Programs ..................................................................... 93<br />
Session 7 Natural Enemies & Biocontrol ................................................................... 103<br />
Session 8 Chemical Ecology, Attractants & Other Control Methods .................................. 117<br />
Workshop Longevity, Ageing & SIT ......................................................................... 133<br />
3
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
POSTER ABSTRACTS .................................................................................................. 139<br />
DATABASES ...................................................................................................... 141<br />
Session 1 Biology, Ecology & Behaviour .................................................................... 145<br />
Session 2 Morphology & Taxonomy .......................................................................... 175<br />
Session 3 Genetics & Evolution .............................................................................. 181<br />
Session 4 Risk Assessment, Quarantine & Post-harvest Treatments .................................. 199<br />
Session 5 SIT Principles & Application ...................................................................... 217<br />
Session 6 Area-Wide & Action Programs ................................................................... 237<br />
Session 7 Natural Enemies & Biocontrol .................................................................... 247<br />
Session 8 Chemical Ecology, Attractants & Other Control Methods ................................... 271<br />
Workshop Longevity, Ageing & SIT ............................................................................. 321<br />
AUTHOR INDEX ........................................................................................................ 325<br />
NOTES ............................................................................................................... 335
Welcome message<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
On behalf of the Organizing Committee I wish to warmly welcome you to the 8th<br />
International Symposium on Fruit Flies of Economic Importance (ISFFEI 2010) in Valencia,<br />
Spain, from 26th September to 1st October 2010. I would like to thank to all those people<br />
who have contributed to the preparation of this Symposium as well as all the sponsors of<br />
this event.<br />
We are living a very exciting scientific period, with considerable advances in the field of<br />
plant protection. The 8th ISFFEI symposium will reveal the increasing revealance of<br />
Tephritid research in the entire globe and the importance of fruit flies in world trade of<br />
fresh fruits. Recent genetic, biochemical, chemical, biological and microbiological<br />
breakthroughs have allowed new insights into the integrated fruit fly management<br />
programs. However, there is a constant need not only to keep aware of and develop new<br />
scientific approaches but also to identify adequate and effective environmentally safe<br />
control strategies for the different tephritids of economic importance. The members of the<br />
Organizing and Steering Committees have build a scientific program which will provide a<br />
balanced overview of advances in Tephritids’ basic and applied research, with special focus<br />
on control programs.<br />
The symposium venue is the Nexus building, the Congress centre of the Universidad<br />
Politécnica de Valencia, that will provide an appropriate and secure setting, with up-todate<br />
technical facilities for the meeting. In addition to an exciting scientific program, you<br />
will benefit from the end of the summer, one of the nicest seasons in Valencia. Moreover,<br />
the cultural background and historical landscape that make of Valencia one of the most<br />
attractive cities in Europe, should contribute towards making this event an unforgettable<br />
moment. We hope that this unique environment will be a perfect time to meet friends and<br />
colleagues, and to establish or reinforce collaborations with peers from elsewhere.<br />
Tephritid researchers from all over the globe have convened in Valencia and the 8th ISFFEI<br />
is already here. I expect that the symposium will be a great success and look forward to<br />
meeting all you during these days.<br />
Alberto Urbaneja<br />
8th ISFFEI Chairman<br />
5
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
8 th ISFFEI Committees<br />
Organizing Committee<br />
Chairman<br />
Dr. Alberto Urbaneja García<br />
Centro de Protección Vegetal y Biotecnología. Instituto Valenciano de<br />
Investigaciones Agrarias (IVIA).<br />
Secretary team<br />
Dr. Beatriz Sabater-Muñoz (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Ing. Rafael Argilés (Medfly SIT working program Spain - TRAGSA, IVIA)<br />
Ms. Tania Navarro (Secretariat Fundación Agroalimed)<br />
Members<br />
Staff<br />
Dr. Luis Navarro (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Dr. Florentino Juste (Director IVIA)<br />
Dr. Pedro Castañera (Centro de Investigaciones Biológicas, CIB CSIC)<br />
Mr. Fernando Hernandez (Fundación Agroalimed)<br />
Dr. Francisco Beitia (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Dr. Josep A. Jacas (Departament de Ciències Agràries i del Medi Natural,<br />
Universidad Jaume I)<br />
Dr. Ferran García Marí (Unidad de Entomología Agroforestal. Instituto<br />
Agroforestal Mediterráneo. Universidad Politécnica de Valencia (<strong>UPV</strong>).<br />
Dr. Jaime Primo (Centro de Ecología Química Agrícola (CEQA), <strong>UPV</strong>)<br />
Dr. Vicente Navarro (Centro de Ecología Química Agrícola (CEQA), <strong>UPV</strong>)<br />
Dr. Joel González (CIB)<br />
Dr. Cristian Vidal-Quist (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Ing. María Juan-Blasco (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Dr. César Monzó (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Dr. Victoria San Andrés (Centro de Protección Vegetal y Biotecnología, IVIA)<br />
Dr. Tatiana Pina (Departament de Ciències Agràries i del Medi Natural, UJI)<br />
Staff for Technical visits<br />
Jaime García de Oteyza (TRAGSA, Medfly mass rearing facility)<br />
Igancio Pla (TRAGSA, Medfly & parasitoids releasing facility)
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Miguel Ángel Utrillas (TRAGSA, Medfly & parasitoids releasing facility)<br />
Marta Briasco García (TRAGSA, Medfly & parasitoids releasing facility)<br />
Carles Tur Lahiguera (TRAGSA, Medfly & parasitoids releasing facility)<br />
Teresa Navarro Navarro (TRAGSA, Medfly mass rearing facility)<br />
Marta Martínez Gonzalvo (TRAGSA, Medfly & parasitoids releasing facility)<br />
Antonio Polido (TRAGSA, Medfly mass rearing facility)<br />
Luis Villanueva Gallent (TRAGSA, Medfly mass rearing facility)<br />
Germán Ceballos Chan (TRAGSA, Medfly mass rearing facility)<br />
Scientific Committee<br />
Dr. Alberto Urbaneja<br />
Dr. Beatriz Sabater-Muñoz<br />
Dr. Vicente Navarro<br />
Dr.Francisco Beitia<br />
Dr. Josep Jacas<br />
Dr. Ferran García Marí<br />
Mr. Rafael Argilés<br />
Dr. Brian Barnes<br />
Dr. Anna Malacrida<br />
Dr. Sunday Ekesi<br />
Dr. Nancy Epsky<br />
Dr. Yoav Gazit<br />
Dr. Nikos Kouloussis<br />
Dr. Pablo Liedo<br />
Dr. Aldo Malavasi<br />
Dr. Robert Mangan<br />
Dr. Pablo Montoya<br />
Dr. Serge Quilici<br />
Dr. Monica Spinetta<br />
Dr. Roberto Zucchi<br />
Dr. Kenneth Bloem<br />
Dr. Nicanor Liquido<br />
7
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Fruit Fly Steering Committee (as in June 2008)<br />
Chairman<br />
Dr. Brian Barnes<br />
ARC Infruitec-Nietvoorbij Fruit, Vine & Wine Institute, Stellenbosch<br />
Members<br />
Sunday Ekesi (International Centre of Insect Physiology and Ecology (ICIPE), Nairobi,<br />
KENYA)<br />
Nancy Epsky (Subtropical Horticulture Research Station, USDA ARS, Miami, USA)<br />
Yoav Gazit (The Israel Cohen Institute for Biological Control, PPMB – Citrus Division,<br />
Rishon Le-Zion, ISRAEL)<br />
Nikos Kouloussis (Aristotle University of Thessaloniki, School of Agriculture, 54124<br />
Thessaloniki, GREECE)<br />
Pablo Liedo (El Colegio de la Frontera Sur (ECOSUR), Tapachula, Chiapas, MEXICO)<br />
Anna Malacrida (Università di Pavia. Dipartimento di Biologia Animale, ITALY)<br />
Aldo Malavasi (Moscamed Medfly Program, Sao Francisco Valley, Bahia, BRAZIL)<br />
Robert Mangan (Crop Quality and Fruit Insect Research, USDA ARS, Weslaco, Texas)<br />
Pablo Montoya (Programa Moscafrut, SAGARPA-IICA, Tapachula Chiapas, MEXICO)<br />
Serge Quilici (CIRAD Réunion, Pôle de Protection des Plantes, La Réunion, FRANCE)<br />
Pedro Rendon (USDA/APHIS/PPQ, Methods Development Section, Guatemala City)<br />
Catherine Smallridge (South Australian Research and Development Institute,<br />
Adelaide, South Australia, AUSTRALIA)<br />
Mónica Spinetta (Servicio Nacional de Sanidad y Calidad Agroalimentaria (SENASA)<br />
de Argentina, Programa Nacional de Control y Erradicación de las<br />
Moscas de los Frutos, Buenos Aires, ARGENTINA)<br />
Michael Stefan (Fruit fly Exclusion and Detection Programs, PPQ, APHIS, Riverdale)<br />
Roberto Zucchi (University of Sao Paulo (USP/ESALQ), BRAZIL)
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Acknowledgement to Official Organizing & Supporting<br />
Institutions<br />
Organizing Institutions:<br />
o Instituto Valenciano de Investigaciones Agrarias (IVIA)<br />
o Universitat Jaume I (UJI)<br />
o Universidad Politécnica de Valencia (<strong>UPV</strong>)<br />
o The Fundación Agroalimed<br />
The Organizing Committee would like to give thanks to the following Spanish and<br />
international institutions which had provided funds to develop this event:<br />
o Generalitat Valenciana (GVA ) project AORG/2010/031<br />
o Instituto Nacional de Investigaciones y Tecnología Agraria y Alimentaria (INIA)<br />
project AC2009-00055-00-00<br />
o Universidad Politécnica de Valencia (<strong>UPV</strong>) project 2473<br />
o The Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture<br />
9
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Acknowledgement to Sponsors & Exhibitors<br />
In the following pages delegates could read about our valuable sponsors and<br />
exhibitors.<br />
Sponsors:<br />
� Platinum: SYNGENTA<br />
� Golden: BAYER CROPSCIENCE, CERTIS and DOWAGROSCIENCES<br />
� Silver: FRUIT FLY SERVICES SL.<br />
� Bronze: BIAGRO SL.<br />
Exhibitors:<br />
� AGRISENT<br />
� BIOIBÉRICA<br />
� COLTEC<br />
� FRUIT FLY SERVICES SL.<br />
� MUBARQUI Servicios Aéreos<br />
� SCIENTIFIC ADVISORY SERVICES Pty. Ltd.<br />
� SUTERRA<br />
� SYNGENTA<br />
The Organizing Committee would like to give all them the thanks for their support<br />
to this event.<br />
Thank you!
About ISFFEI<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Initiated in 1982, the quadrennial fruit fly symposium is being well established as a<br />
key meeting for bringing together scientists and fruit fly workers from all over the<br />
world.<br />
The purpose of this Symposium is to bring together academic, industry, research and<br />
action people from the all over the world to discuss the various aspects of fruit flies<br />
and recent developments related to Pest Management Strategies.<br />
The meeting will encourage cooperation between participants, contribute to spread<br />
new findings, promote their practical use and highlight research needs as it has<br />
been done n the previous symposia.<br />
Past Symposia:<br />
• The First International Symposium on Fruit Flies of Economic Importance, Athens,<br />
Greece, November 16-19, 1982.<br />
• The Second International Symposium on Fruit Flies of Economic Importance, Crete,<br />
Greece, 16 - 21 September 1986.<br />
• The Third International Symposium on Fruit Flies of Economic Importance,<br />
Antigua, Guatemala, 14 – 20 October 1990.<br />
• The Fourth International Symposium on Fruit Flies of Economic Importance, Sand<br />
Key, Florida, USA, 5 – 10 June 1994.<br />
• The Fifth International Symposium on Fruit Flies of Economic Importance, Penang,<br />
Malaysia, 1 – 5 June 1998.<br />
• The Sixth International Symposium on Fruit Flies of Economic Importance,<br />
Stellenbosch, South Africa, 6 – 10 May 2002.<br />
• The Seventh International Symposium on Fruit Flies of Economic Importance,<br />
Salvador, Bahia, Brazil, 10 – 15 September 2006.<br />
11
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
General information<br />
Congress language<br />
The official language will be English. However, due to legal requirements, our<br />
government representatives will use one of the two official languages in Valencia,<br />
Spanish or Valencian with English simultaneous translation for the Opening<br />
ceremony.<br />
Congress technical organizers<br />
Grupo Pacífico<br />
Paseo General Martínez Campos, 44, 1º<br />
28010 Madrid - SPAIN<br />
Tel. (34) 913.836.000 ext. 127<br />
Fax. (34) 913.023.926<br />
isffei2010@pacifico-meetings.com; cherrera@pacifico-meetings.com;<br />
broca@pacifico-meetings.com<br />
Congress venue<br />
The Nexus building (6G in the map) of the Universidad Politécnica de Valencia<br />
(<strong>UPV</strong>) harbors the Congress Centre of this Campus and will provide an appropriate<br />
and secure setting, with up-to-date technical facilities, to allow the perfect<br />
development of the Symposium program.<br />
This building can be reached from entry “L” or “K”, and is 1-2 minutes walking from<br />
Galileo Galilei Hall of Residence (6B in the map).<br />
The tram is running all along the <strong>UPV</strong> Campus, with four stops marked with a white<br />
“m” in a red circle in the map. The nearest tramp stop to the Symposium venues is<br />
“Tarongers” (which means orange trees in Valencian).<br />
The <strong>UPV</strong> Campus map is on page 18
Registration desk & Technical secretariat<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
The 8 th ISFFEI technical secretariat is formed by personnel of Grupo Pacífico. The<br />
technical secretariat will be open for on-site registrations and for information about<br />
the Symposium, at the Symposium hours.<br />
The registration desk will be located in the main entrance hall level, please follow<br />
indications or ask Organizers Staff (easily to identify by their T-shirts and badges).<br />
Internet access & Stations<br />
All delegates will be provided with a personal key code (username and password) to<br />
access the wireless net. Internet stations and an area available to delegates to send<br />
and receive email messages free of charge will be located in the first floor.<br />
Shuttle services & Public transport in Valencia city<br />
A shuttle transport will operate between the appointed NH hotels and the ISFFEI<br />
venue as per the schedule on Program. Buses will depart twice in the morning and<br />
after all conferences with 20 minutes of difference between trips. More detailed<br />
timetable will be set up in the NH hotel hall.<br />
Shuttle buses will stop at the “L” entrance of the <strong>UPV</strong> Campus.<br />
Delegates hosted in other hotels are aimed to be in time in the NH hotel hall or have<br />
their own transfer to the <strong>UPV</strong> Campus.<br />
Valencia city transport guides will be provided to all the delegates within the 8 th<br />
ISFFEI bags and have been provided by the Valencia convention bureau. Other hotels<br />
have also information about public transport from any point of Valencia city to the<br />
ISFFEI venue.<br />
Accompanying persons<br />
Accompanying persons who are registered as such will receive a name badge and<br />
tickets for all the ceremonies. Accompanying persons holding the ID badges are<br />
entitled to travel on the official shuttle transportation, attend the opening<br />
ceremony, welcome reception and closing dinner, visit the exhibition hall and<br />
participate on the Accompanying Persons City Tour on Tuesday 28 th September from<br />
9:00 to 13:00.<br />
City Tour will depart at 9:00 from NH Las Artes I hotel.<br />
13
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Speakers, audio visual & Technical desk<br />
World class audio-visual and technical services will be provided by an expert team of<br />
<strong>UPV</strong>, which will be located in the main entrance hall level. Please, follow<br />
indications or ask Organizing Staff.<br />
All presentations are to be computer generated, mainly with MS-PowerPoint (v2003-<br />
07) or compatible software, and provided as .ppt or .pps files. Speakers are required<br />
to report to this desk with their CD’s, memory sticks (USB) or laptops with at least<br />
24h prior to making their presentation. At Session breaks, presenters will be able to<br />
check the performance of their presentation with the hardware of the Auditorium.<br />
First day presenters are encouraged to report to the centre well in advance of their<br />
presentation times to avoid queues and possible delays in the Symposium timetable.<br />
All presentations should be uploaded to the Auditorium central computer. Own<br />
computers will not be admitted as source for the presentation in the Auditorium,<br />
the Organizer cannot guarantee projection of presentations that are handed in this<br />
way.<br />
If you didn’t submit a pdf copy of your talk by 24th September, please refer also to<br />
this desk to upload it for its inclusion in the USB key with all the information of the<br />
8th ISFFEI. You can upload a slightly modified version of your talk protecting your<br />
valuable data, but please give the opportunity to all delegates to keep a copy of the<br />
main ideas of your work. This USB key will be distributed among all participants.<br />
A presentation certificate will be delivered to the presenting delegate after talk in<br />
the Speaker Desk. Only one certificate, listing all the contributing authors, will be<br />
distributed for each talk.<br />
Poster presenters<br />
A Poster Information desk will be located within Poster room number one in the<br />
main entrance hall level. In this desk you will find your poster board number and<br />
double side tape to affix the poster to the poster board. Posters should be mounted<br />
on the poster boards early on Monday September 27th and will be hold during the all<br />
week. Posters must be removed on Thursday 30th September at 6 pm. Any poster<br />
not removed will be taken down by the Organizers and stored until Technical visit.<br />
Poster not collected during Technical visit will be discarded.<br />
If you did not submit a pdf copy of your poster by 24th September, please refer to<br />
the audio-visual desk to upload it for its inclusion in the USB key with all the<br />
information of the 8th ISFFEI. This USB key will be distributed among all<br />
participants.<br />
A presentation certificate will be delivered to the presenting delegate after poster<br />
holding in the Poster Desk. Only one certificate, listing all the contributing authors,<br />
will be distributed for each poster.
Abstracts & CD provided<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
All talk and poster abstracts are included in this book as well as in the CD within the<br />
delegate bag.<br />
Special Events<br />
Welcome Reception: Sunday 26 th September at NH Las Artes I hotel<br />
Opening Ceremony: Monday 27 th September, at the Auditorium, by the Honourable<br />
Consellera of Agriculture of Valencia.<br />
Closing Ceremony: Thursday 30 th September, at the Auditorium.<br />
Farewell dinner: Thursday 30 th September, at the City of Arts & Science.<br />
Technical visit: Friday 1 st October, see schedule for further information.<br />
Bids for the 9 th ISFFEI: Tuesday & Wednesday, by the Steering Committee.<br />
Coffe breaks & Lunch<br />
Breaks are planned as in the 8 th ISFFEI schedule and will take place in a big top tentlike<br />
structure located outdoors at 200 meters from the building entrance.<br />
Morning coffee breaks will take place in this tent, whereas in the afternoon, a<br />
coffee station will be available in Poster room no.1 close to the Exhibition hall, just<br />
on the left side of the Auditorium.<br />
Exhibition<br />
The exhibition hall will be located in the main entrance hall level, and will be open<br />
in the same timetable as the Symposium.<br />
A list of participating Exhibitors appears in the following pages of this book and all<br />
delegates are encouraged to visit them.<br />
15
SPONSORS
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.
Syngenta Crop Protection<br />
We bring plant potential to life<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Syngenta is one of the world’s leading companies with more than 25,000 employees<br />
in over 90 countries dedicated to our purpose: Bringing plant potential to life.<br />
Our Crop Protection and Seeds products help growers increase crop yields and<br />
productivity. We contribute to meeting the growing global demand for food, feed<br />
and fuel and are committed to protecting the environment, promoting health and<br />
improving the quality of life.<br />
Syngenta offers a uniquely broad product portfolio leader in crop protection and<br />
high-value commercial seeds, World-class science (leader in research and<br />
development with $1 billion R&D investments in 2009 and 5,000 people in R&D<br />
around the world), global reach and experience (over 25,000 employees in more<br />
than 90 countries) and commitment to working with customers (tailoring solutions to<br />
individual needs).<br />
Major product brands include Herbicides control weeds that compete with crops for<br />
light and nutrients, Fungicides to prevent and cure fungal disease, Insecticides to<br />
control pests which reduce yields by damaging crops, Seed Care products for<br />
protecting young plants against insects and disease and Seeds for corn, soybean,<br />
sugar beet, cereals, oil crops; vegetables and flowers.<br />
Since 2006, Syngenta commercializes in Spain and other Mediterranean countries<br />
Adress ® , a new method to control Med Fly based in the reduction of fly populations.<br />
Adress ® system was developed together by Syngenta and the University of Valencia<br />
helps farmers to control this important pest with technology respectful with the<br />
environment and beneficial insects and it is perfectly compatible with other<br />
methods such as sterile insect technique (SIT).<br />
Adress ® has been tested in many trials and different conditions in citrus, top fruits,<br />
table grapes and tropical fruits, including the complete protection of the whole<br />
Hierro Island (Canary Islands).<br />
For more information please visit:<br />
www.syngenta.com<br />
19
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Bayer CropScience aims to contribute to sustainable agriculture, which means<br />
achieving a harmonious balance between economic success, environmental<br />
responsibility and social acceptability.<br />
With its considerable investment in R&D, Bayer CropScience is able to offer<br />
technologies and agricultural solutions that guarantee crops, reduce losses, improve<br />
the quality of produce from the fields and optimise the use of natural resources,<br />
contributing to meeting world food and animal feed requirements.<br />
Our company is one of the global leaders in the supply of innovative products, not<br />
only of insecticides and herbicides, fungicides and seed treatments but also of<br />
integral solutions and services.<br />
With respect to insecticides, Bayer CropScience has always had a high profile in the<br />
control of Ceratitis capitata, being the company that developed Fention (Lebaycid)<br />
that made it possible to control this pest for many years. Our company continues to<br />
conduct research to develop new solutions that will meet the demands of<br />
sustainable agriculture, in agreement with the demands we place on ourselves.<br />
In line with this strategy, Bayer CropScience will be launching an innovative<br />
solution as part of its Mass Trapping solutions this year. This is Decis Trap, which<br />
will enable us to contribute to the necessary modernisation required by all the<br />
Ceratitis control systems. We are also taking initial steps in cooperation with various<br />
official bodies, to conduct research into new solutions that will help to provide an<br />
integrated response for the correct management of this pest, which is of recognised<br />
importance to agriculture in the Mediterranean area.<br />
For more information please visit:<br />
http://www.bayercropscience.es/<br />
http://www.bayercropscience.com/
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Since its launch in 2001, Certis Europe has developed and grown steadily. It has<br />
expanded its activities both geographically and in terms of technologies and product<br />
portfolio. Unique among suppliers to the horticultural and specialty crops sectors,<br />
Certis holds true to its original objectives to provide solutions for growers that take<br />
account of market demands and pressures, as well as crop production challenges.<br />
Constantly at the forefront of innovation, Certis Europe uses a number of new<br />
technologies in its product development to promote sustainability. Important<br />
amongst these are the beneficial insects, including predators and parasitoids,<br />
biopesticides and other compatible products used in Integrated Pest Management<br />
(IPM) programmes to provide effective and sustainable pest control.<br />
Virus-, bacteria- and pheromone-based products also offer important additions to<br />
the armoury and have proved a valuable asset in the development of integrated<br />
control strategies in both fruit and vegetables. A major advantage for all these<br />
products is that they leave no residues in fruit and have a zero harvest interval.<br />
Certis can thus provide growers with solutions to produce healthy, clean and safe<br />
crops, with protection available right up to harvest.<br />
For more information please visit:<br />
http://www.certiseurope.com/<br />
21
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Ceratitis capitata (Wiedemann), courtesy of USDA-ARS)<br />
GF-120 ® Fruit Fly Bait.<br />
� Manage and eradicate populations of fruit flies quickly and easily using GF-<br />
120 ® .<br />
� GF-120 ® is a fruit fly bait consisting of natural feeding attractants with the<br />
active ingredient spinosad.<br />
� All ingredients are optimized for attraction of multiple tephritid species including:<br />
Mediterranean fruit fly, melon fruit fly, Oriental fruit fly, olive fruit<br />
fly, cherry fruit fly, walnut husk fly<br />
� Attract and control male and female of major fruit fly species of economic<br />
importance.<br />
� It has been used globally to eradicate and/or manage multiple species of fruit<br />
flies<br />
� Does not disrupt beneficial insect populations<br />
� Registered to be used in organic agriculture<br />
� For more information please visit: http://www.dowagro.com<br />
® Trademark of Dow AgroSciences LLC. Always read and follow label directions.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FRUIT FLY SERVICES, SL born as result of the union of two pioneer companies in<br />
rational control of agriculture pests by means of respectful systems with the<br />
environment, PROBODELT and OPENNATUR.<br />
It born with the intention to become a referent in pest control of different dipterans<br />
pests that affect nowadays at Spanish crops, as well as new species that could<br />
introduced and could be potentially harmful to our crops.<br />
PROBODELT, with a wide experience in pest control, was created with the target of<br />
design and execute rational pest control programmes based in integration of<br />
different techniques with easy application systems. Has been pioneer in introduction<br />
of mass trapping technique at commercial level against rice borer (Chilo<br />
supressalis). It’s specialized in insect traps manufacturing and patents makes it a<br />
reference in pest control sector.<br />
OPENNATUR, specialized in pest control by means environmentally respectful<br />
methods, was created as well as PROBODELT with the target of design and execute<br />
pest control programmes. Since the beginning, its bet was strong in I+D+I purposes,<br />
its allows to be a reference in the pest control sector with support of other different<br />
techniques as monitoring, mass trapping, and mating disruption.<br />
FRUIT FLY SERVICES unite in only one company the experience, knowledge and<br />
resources that had allows in a short period of time to become a reference company<br />
for pest control of Ceratitis capitata and Bactrocera oleae.<br />
For more information please visit:<br />
http://www.opennatur.com/<br />
www.probodelt.com<br />
23
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
BIAGRO S.L. is a Spanish company operating in Vegetable Nutrition and Integrated Pest<br />
Management. Founded by Mr. Antonio Quesada in 1982, the company started operating as a<br />
consultant of big fruits and vegetables producers with a branch society called Consultant<br />
Centre of Plant Nutrition, whose funcion was the nutritional diagnosis, fertilisation<br />
programs, observation and checking of agricultural industries and advice on growing<br />
techniques. After 16 years of operation, BIAGRO SL. produces its own fertilizers, correctors<br />
and natural fungicides, and distributes insect pheromones and IPM products from a well<br />
know suppliers. BIAGRO S.L. believes in continuos and accelerated growth through its own<br />
I+D department and through strategic research alliances with academic partnerships. At the<br />
same time, the Company is involved and cooperate with the Ministry of Agriculture in<br />
scientific programs to develop new IPM systems with pheromones and natural products.<br />
BIAGRO SL. is a charter member of AEFA (Spanish Manufacturer Agronutrients Association).<br />
It forces us to make high quality products, adjusted to the Spanish and European rules.<br />
BIAGRO SL. has achieved the quality procedure ISO 9001.<br />
Recently and regarding IPM of Tephritids in Spain BIAGRO SL. has developed STARCE a liquid<br />
bait with very interesting results (80% of medfly females captures vs 20% of males), and<br />
continues with the distribution of M3 traps (Green Trading Co.) for lure and kill medfly. M3<br />
traps are being recommended by the Department of Agriculture of the Valencia Region<br />
Government.<br />
Learn more at:<br />
www.biagro.es
EXHIBITORS
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
List of Exhibitors:<br />
� AGRISENT<br />
� BIOIBÉRICA<br />
� COLTEC<br />
� FRUIT FLY SERVICES SL.<br />
� MUBARQUI Servicios Aéreos<br />
� SCIENTIFIC ADVISORY SERVICES Pty. Ltd.<br />
� SUTERRA<br />
� SYNGENTA
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Address: AGRISENT DE MEXICO SA de CV. Canales 170 Col. Alianza. Entre<br />
Av. Del Maestro y Sn Carlos. H. Matamoros Tamp. MEXICO<br />
Telephone: 868 1 49 10 94<br />
Email: ehcd@aol.com<br />
Web site: http://www.agrisent.com<br />
Information: Agrisent de México SA de CV, started in 1992. Is a líder in supply of<br />
diet ingredients (as corn cob, torula yeast), materials and other<br />
equipements for rearing beneficial insects including those used in SIT. It<br />
provides support to international, federal or state agencies and to the<br />
private sector, regarding the development, knowledge and production of<br />
beneficial insects and those used in SIT. In cooperation with several<br />
Mexican government agencies and SAGARPA, has developed and tested<br />
new products for monitoring and control of insects. The latest is the ECO-<br />
TML Plug Phermone (Ecological Trimedlure Capsule). Ingredients for<br />
larval diets: corn cob meal, torula<br />
Agrisent de México SA de CV, fue incorporada en 1992. Líder en el<br />
suministro de ingredientes para dietas larvarias (harita de olote, levadura<br />
de torula), materiales y equipos para los Mercado de insectos benéficos y<br />
Sistemas Administrativos Integrados para el control y erradicación de<br />
plagas a Agencias inter-nacionales y Agrícolas gubernamentales<br />
federales, estatales y al sector privado en apoyo al esfuerzo, al estudio,<br />
desarrollo y producción de insectos benéficos. En cooperación con<br />
algunas agencias estatales Mexicanas y SAGARPA, ha desarrollado y<br />
probado, nuevos productos para el monitoreo y control de insectos. El<br />
más nuevo es el ECO-TML Phermone Plug, (Capsula de Trimedlure<br />
Ecológica).<br />
27
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Address: BIOIBERICA SA. Plaza Francesc Macià, 7. 08029 Barcelona, Spain.<br />
Telephone: (+34) 93 490 49 08 FAX (+34) 93 490 97 11<br />
Email: rplana@bioiberica.com<br />
Web site: http://www.bioiberica.com; www.ceratrap.com; www.plantstress.net<br />
Information: Since 1975 Bioiberica has specialized in the research, manufacturing and<br />
marketing of biomolecules extracted by enzymatic hydrolysis for the pharmaceutical,<br />
veterinary and agricultural industry.<br />
A firm commitment and extensive knowledge has consolidated Bioiberica as<br />
a leading producer of active ingredients such as amino acids, characterized<br />
peptides and proteins. The protein based formulations developed by<br />
Bioiberica are specific biological attractants that are highly selective against<br />
insects and harmless to beneficials. Their high attraction power allows controlling<br />
efficiently target pests without adding any insecticide.<br />
Plant Physiology Department offers a range of Biological Attractants that are<br />
effective and completely environmentally friendly solutions that work against<br />
different pests and fruit flies.<br />
Thanks to this, BIOIBERICA - Plant Physiology has contributed to significant<br />
improvements in agriculture, offering specific solutions for all kinds of<br />
plant stresses. Our products are commercialized in more than 50 countries and<br />
we produce more than 80 references.<br />
The R&D Department and its skilled staff of Agronomists, Biochemists and<br />
Pharmacists works on the development and preparation of new products in<br />
close collaboration with various Universities and International & National<br />
Scientific Institutions.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Address: COLTEC, COMERCIALIZADORA AGROTECNOLOGICA S.A.<br />
8 Calle 11-13 zona 1 Oficina #108, Guatemala, GUATEMALA.<br />
Telephone - Fax: (502) 2366 0937 - (502) 2366 0936<br />
Email: ebusto@coltec.com.gt<br />
Web site: http://www.bioiberica.com<br />
Information: COLTEC is a specialized company for the mass rearing, with nutrients<br />
for the diet, Yeasts, Proteins, Complete Diets, equipment, Trays and<br />
baits Atrapol. We represent Lallemand and Molded Fiber Glass<br />
Company. Fiber Glass Trays<br />
www.mfgtray.com<br />
29
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Address: SERVICIOS AEREOS MUBARQUI. Blvd. Enrique Cárdenas Gonzáles<br />
1356 fracc. Los arcos. Ciudad Victoria Tamaulipas México C.P.<br />
87040. MEXICO<br />
Telephone - Fax: 834-3164921 - 834-3146841<br />
Email: rakoperations@yahoo.com.mx<br />
Web site: http://201.101.7.234/empresa/<br />
Information: SERVICIOS AEREOS MUBARQUI is a specialized company for<br />
release of sterile insects used in SIT programs. Developing alternatives<br />
for eclosion of sterile insects to allow the development of more outcompeting<br />
insects being a key factor for SIT.<br />
SERVICIOS AEREOS MUBARQUI has developed aerial release automatic<br />
machines with intelligent refrigerated units, and MACX system<br />
(a real time release analysis system), which jointly allows a follow up of<br />
each sterile mexfly release in terms of quality controls.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Address: Scientific Advisory Services Pty Ltd. PO box 1056. Tully, Queensland<br />
4854, AUSTRALIA<br />
Telephone – Fax : 61 0417 644 660 61 7 4065 4991<br />
Email: info@saspl.com.au<br />
Web site: http://www.saspl.com.au/<br />
Information: SAS has published a number of high quality wall posters on fruit flies<br />
(Tephritidae) of Australia and the World, as well as bell injection of<br />
bananas and insect pests and beneficials of bananas and papaws in Australia.These<br />
posters are valuable aids for primary producers as well as<br />
government quarantine agencies both in Australia and overseas.<br />
Scientific Advisory Services has recently formed a unique partnership<br />
with Starr, an exciting young Brisbane artist to produce a unique range<br />
of insect paintings.<br />
SAS offers also insects embed into resin, as key rings. Please have a<br />
look in the trials with fruitflies as paper-holders just made a few weeks<br />
ago!<br />
Some of the Starr insect paintings, beautiful!<br />
31
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Address: SUTERRA Corporate. 20950 NE Talus Place Bend Oregon 97701,<br />
USA<br />
Telephone – Fax : 541-388-3688 541-388-3705<br />
Email: david.loughlin@suterra.eu; monica.dominguez@suterra.eu<br />
Web site: http://www.suterra.com/<br />
Information: Suterra is dedicated to aiding farmers in finding workable solutions by<br />
researching, developing and commercializing alternative IPM solutions<br />
using naturally occurring attractants and semiochemical compounds to<br />
protect crops such as pome fruit, stone fruit, citrus and vegetables from<br />
insect attack. Headquartered in Bend, Oregon and with regional offices<br />
in Spain, Italy, Mexico and Argentina, Suterra is a market leader in the<br />
manufacture and supply of a broad range of IPM tools for a variety of<br />
crop pests. Suterra's pheromone and attractant based control products<br />
are already registered in many global markets in the Americas, Europe<br />
and the Southern Hemisphere and Suterra's reputation and expertise is<br />
recognized throughout the world by university entomologists,<br />
government agencies and farming organizations. For fruit flies, the<br />
range extends beyond attractants for monitoring males and females and<br />
now includes effective attractant solutions for use in area-wide, mass<br />
trapping control campaigns and the novel attract and kill Magnet<br />
tools for both Olive Fly and Mediterranean Fruit Fly: Magnet OLI<br />
and Magnet MED
Program at glance
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
26/09/10<br />
Sunday<br />
27/09/10<br />
Monday<br />
08.00 Registration and<br />
Poster holding<br />
08.30<br />
09.00 Opening ceremony<br />
09.30 Plenary confer-<br />
10.00<br />
ence<br />
28/09/10<br />
Tuesday<br />
10.30 Coffee break / Poster session<br />
11.00<br />
11.30<br />
12.00<br />
12.30<br />
13.00<br />
13.30<br />
14.00<br />
14.30<br />
15.00<br />
15.30<br />
16.00<br />
29/09/10<br />
Wednesday<br />
Plenary conferences<br />
30/09/10<br />
Thursday<br />
Session 8 Session 7 Session 4<br />
Session 3 Session 8 Session 7 Session 5<br />
Lunch<br />
Coffee break /<br />
Poster session<br />
Coffee break / Poster session<br />
Session 1<br />
Session 2 Session 5<br />
16.30 Session 6 /<br />
General discus-<br />
17.00<br />
Workshop Break sion<br />
General discussion<br />
17.30<br />
18.00<br />
18.30<br />
Registration<br />
to the 8th<br />
ISFFEI at<br />
NH Las<br />
General discussion<br />
Session 1<br />
Meeting<br />
TEAM<br />
of<br />
Closing session<br />
Poster removal<br />
19.00<br />
Artes I hotel<br />
General dis-<br />
19.30<br />
cussion<br />
20.00<br />
20.30<br />
21.00<br />
21.30<br />
Welcome<br />
reception at<br />
NH Las<br />
Artes I hotel<br />
Farewell dinner<br />
01/10/10<br />
Friday<br />
Exit from hotels<br />
Technical visit<br />
SIT facilities &<br />
IVIA<br />
Transfer to<br />
hotels
Sessions<br />
Session 1: Biology, Ecology & Behaviour<br />
Session 2: Morphology & Taxonomy<br />
Session 3: Genetics & Evolution<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Session 4: Risk Assessment, Quarantine & Post-harvest Treatments<br />
Session 5: SIT Principles & Applications<br />
Session 6: Area-wide & Action Programs<br />
Session 7: Natural Enemies & BioControl<br />
Session 8: Chemical Ecology, Attractants & other Control Methods<br />
Plenary Sessions<br />
Monday 27 th<br />
Insecticide resistance in fruit flies: the case of malathion resistance in Spanish<br />
populations of Ceratitis capitata by Dr. Félix Ortego (CIB CSIC, Spain)<br />
Tuesday 28 th<br />
A review of recent advances and future prospects in the study of fruit fly (Diptera:<br />
Tephritidae) - host plant relationships by Dr. Martin Aluja (Instituto de Ecología,<br />
Mexico)<br />
Wednesday 29 th<br />
Fruit fly semiochemicals: ‘Discovery and application of attractants and pheromones<br />
in fruit fly control methods’ by Dr. Eric Jang (U.S. Pacific Basin Agricultural<br />
Research Center, USDA-ARS, USA)<br />
Thursday 30 th<br />
In and out of Africa: invasion history of fruit flies on the African mainland (diptera,<br />
tephritidae) by Dr. M. de Meyer (Royal Museum for Central Africa (RMCA),<br />
Entomology Section, Belgium)<br />
35
PLENARY TALKS
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
INSECTICIDE RESISTANCE IN FRUIT FLIES: THE CASE OF MALATHION<br />
RESISTANCE IN SPANISH POPULATIONS OF CERATITIS CAPITATA.<br />
Ortego, Félix*; Couso-Ferrer, Francisco; Arouri, Rabeh; Perera, Nathalie; Beroiz,<br />
Beatriz; Cervera, Amelia; Hernández-Crespo, Pedro & Castañera, Pedro<br />
Centro de Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC),<br />
Departamento de Biología Medioambiental, Madrid, Spain. Email: ortego@cib.csic.es<br />
Fruit flies are considered pests of major economic importance worldwide. A wide range of<br />
management strategies are being implemented for the control of these species, but most control<br />
programs depend on the use of insecticides. Recent studies have reported resistance in field<br />
populations of some species, such as the Mediterranean fruit fly (medfly) Ceratitis capitata<br />
(Wiedemann), the oriental fruit fly Bactrocera dorsalis (Hendel), and the olive fly Bactrocera<br />
oleae (Gmelin). The establishment of resistant populations may become an important economic<br />
problem, particularly if cross resistance is detected in areas where pesticide application is<br />
restricted to a very few active ingredients. The identification of the mechanisms associated to<br />
resistance and the development of tools for the molecular or biochemical detection will be a<br />
key issue to be addressed in order to implement appropriate resistance management strategies<br />
counteracting the emergence and dispersal of resistance.<br />
Resistance to malathion in field populations of C. capitata was first reported by our group in<br />
the Comunidad Valenciana (Spain), where an intensive use of this insecticide was maintained<br />
for several years. A point mutation of the acetylcholinesterase gene (ace2) was elucidated as<br />
the main resistance mechanism, though metabolic resistance mediated by esterases may also be<br />
involved. We have developed a rapid and simple PCR-RFLP diagnostic method for detection<br />
of malathion resistance alleles. Analysis of field populations from different geographical areas<br />
revealed that the resistant allele is widespread all over Spain, whereas it was not detected in<br />
populations from other countries.<br />
Current medfly control in citrus orchards in Spain is mainly based on the use of spinosad and<br />
lamba-cyhalotrin, since malathion was disallowed in the European Union in 2009. The<br />
susceptibility of field populations to these insecticides and the cross-resistance exhibited by the<br />
malathion resistant strain is being evaluated. C. capitata populations tested so far have been<br />
highly susceptible to spinosad. However, remarkable differences in susceptibility to lambdacyhalotrin<br />
have been found when comparing laboratory and field populations. In addition,<br />
selection for resistance to lambda-cyhalothrin or spinosad performed in the laboratory has<br />
resulted in a decreased susceptibility for both insecticides. The mechanisms underlying this<br />
resistance are under study.<br />
Keywords: Medfly, malathion, cross-resistance, lambda-cyhalothrin, spinosad, esterases.<br />
39
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
A REVIEW OF RECENT ADVANCES AND FUTURE PROSPECTS IN THE STUDY OF<br />
FRUIT FLY (DIPTERA: TEPHRITIDAE) - HOST PLANT RELATIONSHIPS<br />
Aluja, Martín*; Birke, Andrea & Guillén, Larissa.<br />
Instituto de Ecología, A.C. Xalapa, Veracruz, Mexico;. Email: martin.aluja@inecol.edu.mx<br />
We provide an overview of recent advances in the study of fruit fly (Diptera: Tephritidae) -<br />
host plant relationships as well as a look into the future of this fascinating research area of<br />
great basic and applied interest. We start by reviewing the evolutionary, ecological,<br />
physiological and behavioral mechanisms related to host finding and utilization. We then<br />
review recent work by us and various other groups worldwide on host plant use patterns,<br />
dwelling on host resistance, fitness costs to the herbivore and the effect of the host on<br />
nutritional and immune reserves and sexual performance of adult flies. Examples used stem<br />
from work with Anastrepha ludens, A. striata, A. fraterculus, Rhagoletis completa and<br />
Ceratitis capitata, among others. We also discuss recent work on host effects on the next<br />
trophic level, using native and exotic braconid parasitoids such as Utetes anastrephae and<br />
Diachamismorpha longicaudata as models to illustrate the phenomenon. Along the way we<br />
discuss the great potential of new research tools such as proteomics in elucidating the fine<br />
details of host plant use by true fruit flies. We finish by discussing recent advances in host<br />
plant status designation; among them the process of determination of the fundamental and<br />
realized host ranges which impinge directly on regulatory procedures related to the export of<br />
fresh fruit worldwide.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FRUIT FLY SEMIOCHEMICALS: DISCOVERY AND APPLICATION OF ATTRACTANTS<br />
AND PHEROMONES IN FRUIT FLY CONTROL METHODS<br />
Jang, Eric.<br />
U.S. Pacific Basin Agricultural Research Center, USDA-ARS, P.O. Box 4459, Hilo, HI 96720. Email:<br />
Eric.Jang@ARS.USDA.GOV<br />
Tephritid fruit flies are some of the most economically important pests of fruits and vegetables<br />
worldwide. As such the discovery and application of semiochemical attractants for detection<br />
and control have had a long history. Early attractants were “discovered” quite by chance but<br />
was quickly followed by both empirical and non-empirical research that lead to the<br />
identification of pheromones, kairomones for some flies. True pheromones have played only a<br />
minor role in control while the “parapheromones” have played a major role, especially for flies<br />
in the family Bactrocera. Fruit flies in the family Anastrepha, Rhagoletis, and Ceratitis have<br />
depended primarily on the application of proteinaceous food-based lures. This is also true of<br />
most of the other (non-Bactrocera) tephritids. Recently, improvements in food-based lures<br />
have resulted in improvements in the application of such attractants for some of the above<br />
species.<br />
The most successful example of the use of attractants for control involves the use of the<br />
parapheromone methyl eugenol for the control of certain Bactrocera fruit flies such as the<br />
oriental fruit fly, B. dorsalis. This technique coined male annihilation or MAT has been the<br />
centerpiece of semiochemical-based control and eradication technologies.<br />
Interestingly behaviorally based methods such as “pheromone disruption” has not been<br />
developed to date; perhaps due to the lack of truly powerful pheromones identified for this<br />
genera. Much of the current research has focused on improvements in attractants through<br />
chemical structure-activity type screening that as resulted in some improvements and well as a<br />
host of new host-based kairomone attractants. Formulation of multiple-component attractants<br />
continues to be a challenge but promise to further improve on the use of semiochemical attracts<br />
for detection and/or control.<br />
41
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IN AND OUT OF AFRICA: INVASION HISTORY OF FRUIT FLIES ON THE AFRICAN<br />
MAINLAND (DIPTERA, TEPHRITIDAE).<br />
De Meyer, Marc *1 ; Ekesi, Sunday 2 ; Virgilio, Massimiliano 1-3 ; Khamis, Fathiya 2 ; Mwatawala,<br />
Maulid 4 & Malacrida, Anna 5<br />
1 Royal Museum for Central Africa (RMCA), Entomology Section, Tervuren, Belgium; Email:<br />
marc.de.meyer@africamuseum.be; 2 International Centre for Insect Physiology and Ecology, Nairobi, Kenya;<br />
3 Royal Belgian Institute of Natural Sciences, Invertebrates Section, Brussels, Belgium; 4 Sokoine University of<br />
Agriculture, Department of Crop Science and Production, Morogoro, Tanzania; 5 Pavia University, Department of<br />
Animal Biology, Pavia, Italy<br />
Recent increases in trans-continental transport of tropical fruits and vegetables as a result of<br />
globalization in trade, as well as human traffic either for leisure (tourism) or business, is<br />
leading to the increased risk of inadvertently introducing alien invasive species to countries or<br />
regions where they not already occur with devastating economic impact if not controlled. Such<br />
human activities cause dispersal of organisms over long distances that could not be covered by<br />
natural means. The introduction of species into a new area can alter successional patterns,<br />
mutualistic relationships, community dynamics, ecosystem function, and resource distribution.<br />
Fruit flies (Diptera, Tephritidae) are a group of organisms that are very susceptible to such<br />
human induced introductions and few insects have greater impact on international marketing<br />
and world trade in agricultural produce than tephritid fruit flies. As important pest species of<br />
several horticultural products like fruits and vegetables, migration and transport of horticultural<br />
produce has lead to (pre-) historic cases of accidental introductions. Although Africa is known<br />
to be the origin of several fruit fly introductions and establishments worldwide (the most<br />
notorious species being the Mediterranean fruit fly, Ceratitis capitata), with the intensification<br />
of fruit trade, the continent has also become highly vulnerable to introduction of alien fruit fly<br />
species. Notable invasive species include Bactrocera zonata, Bactrocera invadens, Bactrocera<br />
latifrons and Bactrocera cucurbitae.<br />
Lately, molecular methodologies is providing useful tools in detecting and analysing alien<br />
introductions. DNA barcoding, using a short gene sequence taken from standardized portions<br />
of the genome, allow recognition of indigenous and alien species even in immature stages that<br />
are inseparable on morphological grounds. This allows quarantine officers and national plant<br />
protection agencies to identify intercepted material during inspections or surveying activities.<br />
Molecular phylogeny indicates the lineages among species, recognizing clusters of closely<br />
related taxa. Combined with host plant data, patterns in host-insect relationships can be<br />
revealed. Population genetics can provide an insight in the macrogeographic population<br />
structure and interrelationships between regions, indicating possible sources of introductions or<br />
migrations. Molecular phylogeography allows a reconstruction of pathways of geographic<br />
range expansion. The application of molecular tools in these fields is demonstrated here with<br />
some examples of fruit fly pests originating from or introduced to the African mainland.
ORAL PRESENTATIONS
Session 1<br />
Biology, Ecology & Behaviour
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
WOLBACHIA IN EUROPEAN, PERSIAN AND RUSSIAN RHAGOLETIS CERASI (DIP-<br />
TERA; TEPHRITIDAE) POPULATIONS.<br />
Rasool, Bilal 1,2* ; Arthofer, Wolfgang 3 ; Schuler, Hannes 2 ; Riegler, Markus 4 ; Mirak, Tohid<br />
N 5 . & Stauffer, Christian 2<br />
1 Nuclear Institute for Agriculture & Biology, Faisalabad, Pakistan, bilalisb2001@yahoo.com; 2 Institute of Forest<br />
Entomology, Boku, University of Natural Resources & Applied Life Sciences, Vienna, Austria. 3 Molecular Ecology<br />
Group, University of Innsbruck, Austria. 4 Centre for Plants and the Environment, University of Western Sydney,<br />
Australia. 5 Cereal Research Department; Karaj, Iran<br />
Background: The European Cherry Fruit Fly, Rhagoletis cerasi, is an established field model<br />
species for multiple Wolbachia infections and cytoplasmatic incompatibility CI. European<br />
populations of R. cerasi were described to be infected with at least five Wolbachia strains<br />
wCer1-5 and wCer2 causes almost complete CI between infected males and uninfected<br />
females. We will compare the wCer infection between R. cerasi specimens collected in 2002<br />
and those collected in 2009. In 2009, also specimens from Persia – the centre of origin of the<br />
host tree Prunus spp. – and from Central Russia were analysed.<br />
Methods: Pupae were collected in diverse areas of Europe, one location close to Teheran (Iran)<br />
and one location in Tomsk (Russia). Specimens were analyzed by sequencing wsp amplicons.<br />
Southern hybridization technique was applied for the detection of the recombinant low titre<br />
strain wCer3.<br />
Results: wCer1 was fixed in all populations whereas wCer2 was present only in South and<br />
Central Europe. The wCer2 spread in Europe from South to North had not changed much from<br />
2002 to 2009 confirming the low migration rate of R. cerasi. The abundance of wCer4 was<br />
homogenous across Iran and Russia and the frequency of wCer5 was highest in the Persian<br />
population. wCer5 showed significant differences in spatial distribution.<br />
Conclusions: No additional wCer strains were detected in Persia. Persia is described to be the<br />
center of origin for Prunus spp.. These centers are assumed to comprise the highest<br />
biodiversity and also genetic diversity. As wCer2 was not detected in Persia, a recent spread of<br />
wCer2 from South Europe is likely. Considering the low migration of R. cerasi and the<br />
presence of wCer1,4,5 in Iran and in the remote and isolated Russian area it is likely that these<br />
strains originate from Persia. From there, R. cerasi spread parallel to its host tree into the<br />
Palearctic region.<br />
Keywords: Rhagoletis cerasi, Wolbachia, cytoplasmatic incompatibility, Prunus, distribution<br />
47
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
INTRA AND INTERSPECIFIC LARVAL COMPETITION BETWEEN CERATITIS<br />
CAPITATA AND ANASTREPHA FRATERCULUS.<br />
Liendo, M. Clara *1,3 ; Devescovi, Francisco 1,3 ; Parreño, María A. 1 ; Milla, Fabián H. 1 ;<br />
Carabajal Paladino, Leonela 1,3 ; Vera, M. Teresa 2,3 ; Segura, Diego F. 1,3 & Cladera, Jorge<br />
L. 1<br />
1 Instituto Nacional de Tecnología Agropecuaria (INTA), Instituto de Genética Ewald Favret (IGEAF); De Los<br />
Reseros y N. Repetto, (1712), Castelar, Buenos Aires, Argentina. Email: mliendo@cnia.inta.gov.ar; 2 Estación<br />
Experimental Agroindustrial Obispo Colombres (EEAOC), Tucumán, Argentina; 3 CONICET.<br />
Background: Ceratitis capitata and Anastrepha fraterculus are two major fruit pests in<br />
Argentina where they are commonly found sharing its host fruit with conspecific and<br />
heterospecific larvae. As larvae are incapable of leaving the fruit, they are forced to share<br />
nutritional resources, and eventually compete for them with other larvae. Larval competition<br />
can occur by direct interference (contest) or exploitation (scramble). The impact of competition<br />
will depend both on the density of larvae within a particular fruit and the relative abundance of<br />
each species. Here we studied both the effects of competing with conspecific and<br />
heterospecific larvae for A. fraterculus and C. capitata.<br />
Methods: Intraspecific competition was analyzed by transferring newly hatched larvae into a<br />
fixed amount of larval diet at increasing densities. Interspecific competition was analyzed by<br />
keeping total density constant and transferring different proportions of larvae of the two<br />
species. We measured percentage of recovered pupae and adults, pupae weight, and larva-pupa<br />
and larva-adult developmental times.<br />
Results: Under intraspecific competition, C. capitata showed a density-dependent reduction in<br />
pupae weight and percentage of recovered pupae. A. fraterculus showed also a density<br />
dependent effect on pupae weight, and a decrease in larva-pupa developmental time.<br />
Comparing the two species, in C. capitata the reduction in pupae weight was more<br />
homogeneous among individuals, whereas in A. fraterculus a majority of pupae showed a<br />
marked decrease in weight and only few pupae showed slight negative effects. Under<br />
interspecific competition, we found that when the relative density of A. fraterculus larvae<br />
increased, the percentage of recovered pupae and the pupae weight increased for C. capitata.<br />
Likewise, A. fraterculus pupae weight increased when the relative density of C. capitata<br />
increased.<br />
Conclusions: Our results suggest that C. capitata larvae facing a shortage of resources develop<br />
a strategy of resource distribution similar to that described as scramble competition, because<br />
the individuals showed a homogeneous impact on their weight. Conversely, A. fraterculus<br />
larvae showed a response similar to that described as contest competition, because as the<br />
number of individuals in the container increases, the negative effect is not the same for every<br />
individual. Finally, we found that C. capitata and A. fraterculus larvae suffer more from<br />
sharing the resource with conspecific larvae than heterospecific larvae.<br />
Keywords: Ceratitis capitata, Anastrepha fraterculus, interspecific competition, intraspecific competition, larval<br />
competition.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECTS OF INTRASPECIFIC COMPETITION ON THE LARVAL DEVELOPMENT AND<br />
PUPAL WEIGHT OF DACINI (DIPTERA: TEPHRITIDAE) INFESTING CUCURBITS IN<br />
LA RÉUNION.<br />
Jacquard, Cathy*; Deguine, Jean-Philippe; Delatte, Hélène; Payet, Christine & Quilici,<br />
Serge<br />
Centre de coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), Département<br />
des Systèmes Biologiques, Saint-Pierre (La Réunion), France. * Postal address: 7 chemin de l’Irat, Ligne Paradis,<br />
97410 Saint-Pierre; E-mail: cathy.jacquard@cirad.fr<br />
Background: In La Réunion, cucurbit crops suffer considerable damage due to fruit fly attacks.<br />
A complex of three species (The Melon fly, Bactrocera cucurbitae, The Ethiopian fly, Dacus<br />
ciliatus, and the Indian Ocean fruit fly, Dacus demmerezi) coexist in the island and can infest<br />
16 different species of Cucurbitaceae. In order to develop sound and sustainable management<br />
methods for these pests, we need to know more about their behavior and ecology. The<br />
objective of this study is to clarify the interaction existing at larval stage, and particularly the<br />
competition which may occur between conspecific larvae in a same fruit. According to earlier<br />
studies, squash (Cucurbita pepo) was chosen for laboratory experiments because it is one of<br />
the most damaged cultivated cucurbit in La Réunion.<br />
Method: To measure the effect of intraspecific competition on the survivorship rate of larvae<br />
and on the pupal mass, infestations of squash were carried out in the laboratory with L1 larvae<br />
of B. cucurbitae, D. ciliatus or D. demmerezi, obtained from lab rearing.<br />
Five levels of infestation were selected e.g. A) One larva (L1) for two grams of squash, B) One<br />
larva/1 g, C) Two larvae/1 g, D) Four larvae/1 g and E) Eight larvae/1 g. Each level was<br />
replicated five times. Four days after infestation, pupae were collected daily, weighed<br />
individually and grouped by weight classes in a same box and when they emerged, the adults<br />
were sexed.<br />
Results: Larvae of B. cucurbitae and D. demmerezi don’t have the same response to an increase<br />
of intraspecific competition in a same fruit. Larvae of the first species show a good<br />
survivorship level (63% for level D) whereas larvae of D. demmerezi are more affected by a<br />
high level of competition (18% for level C). On the contrary, pupal weight of B. cucurbitae<br />
clearly decreases when the number of larvae increases in the fruit while pupal weight of D.<br />
demmerezi, does not seem to be affected by intraspecific competition.<br />
Conclusion: It seems that, in the complex of Dacini attacking cucurbits, the species do not have<br />
the same strategy regarding intraspecific competition. The larvae of B. cucurbitae are able to<br />
share resources and maintain a good survivorship finally giving more adults with low weight.<br />
Conversely, in D. demmerezi larval survivorship is decreased but the pupal weight of survivors<br />
remains high. Further studies will be needed on larval interspecific competition in this complex<br />
of fruit flies, in order to understand the performance of each species in situations of coinfestation.<br />
Keywords: larval intraspecific competition, La Réunion, Dacini, Cucurbita pepo<br />
49
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
OVIPOSITION CHOICE OF BACTROCERA PAPAYAE DREW & HANCOCK (DIPTERA:<br />
TEPHRITIDAE): FULLY-RIPE FRUIT OR UNRIPE FRUIT WITH WOUND ON FRUIT<br />
SURFACE.<br />
Rattanapun, Wigunda* 1 & Amornsak, Weerawan 2<br />
1 Department of Agricultural Technology, Faculty of Technology and Community Development, Thaksin<br />
University, Phatthalung, 93110 Thailand. Email: rwigunda@tsu.ac.th; 2 Department of Entomology, Faculty of<br />
Agriculture, Kasetsart University, Bangkhen Campus, Chatuchak, Bangkok, 10900 Thailand.<br />
Background: The oviposition preference of many species of tephritid fruit fly relies on fruit<br />
firmness. To minimise aculeus wear, female tephritid flies are thought to prefer to oviposit in<br />
fruit wounds or softer exopericarp of ripe and fully-ripe fruits over harder exopericarp of<br />
unripe fruit. The fully-ripe fruit was suitable for larval growth and preferred for oviposition by<br />
female fly of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) more than unripe fruit, soft<br />
exocarp of fully-ripe fruit appeared to be the limiting factor of oviposition, with few successful<br />
ovipositions recorded in all studies, and very few in unripe fruit. In this study, we asked<br />
whether female flies that were exposed to both fully-ripe fruit without wound and unripe fruit<br />
with wound on fruit surface, would choose fully-ripe fruit for oviposition or not. The result of<br />
this study can enhance our understanding of host utilization by female fly.<br />
Methods: A choice experiment was conducted to determine the behavior of individual female<br />
Bactrocera papayae Drew & Hancock (Diptera: Tephritidae). Both fully-ripe mango without<br />
wound and unripe mango with wound on fruit surface of mango, Mangifera indica,<br />
Namdorkmai varity (Anacardiaceae) were offered simultaneously in a 30×30×30 cm<br />
observation cage. An individual, 21–22-day-old, mated female fly was released in an<br />
observation cage. Fruit fly behaviors observed and recorded were: (i) duration of fly visits to a<br />
fruit, (ii) number of attempted ovipositions (unsuccessful penetration), and (iii) duration of<br />
successful oviposition events. Observations were done from 09:00–15:00 hours. Twenty<br />
single-fly replicates were conducted. At the end of the day, female flies were dissected to<br />
check whether eggs were presented in their ovaries. All fruits that female flies had laid eggs<br />
into were dissected for number of egg count.<br />
Results: The total duration of visits differed significantly between the stages of ripening (t-test:<br />
t = 15.010, d.f. = 38, P < 0.0001), with unripe fruits being visited in longer and fully-ripe fruit<br />
significantly less. There were also significant differences in the number of attempted<br />
ovipositions between the various fruit ripening stages, with fewer attempted ovipositions into<br />
fully-ripe fruit and higher penetration into unripe fruit (t-test: t = 6.696, d.f. = 38, P < 0.0001).<br />
All female flies could not oviposit into full-ripe fruits. The average time of successfully<br />
oviposition duration into wound on unripe fruit of female flies is 35.05 ± 1.52 minutes.<br />
Conclusions: The results of this study indicated that fruit wound play an important role in the<br />
oviposition determination of female fly B. papayae. Female B. papayae chose unsuitable host<br />
that she could easier oviposit than nutritious host which was hard to oviposit. Thus, it is<br />
difficult to assume that oviposition preference of female flies relates solely to the performance<br />
of their larvae.<br />
Keywords: Bactrocera papayae, oviposition preference, mango, ripening.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PHENOTYPIC PLASTICITY OF THERMAL TOLERANCE IN ADULT CERATITIS<br />
CAPITATA AND CERATITIS ROSA (DIPTERA: TEPHRITIDAE).<br />
Nyamukondiwa, Casper * & Terblanche, Jonh S.<br />
Department of Conservation Ecology and Entomology, Stellenbosch University, Private Bag X1, Matieland, 7602,<br />
South Africa. Email; tetra@sun.ac.za.<br />
Background: The invasion success of Ceratitis capitata relative to its congener, Ceratitis rosa<br />
probably stems from physiological, morphological and behavioural adaptations which enable<br />
them to survive in different habitats. However, few studies have investigated thermal tolerance<br />
as a means to explain biogeographic differences or likely invasion potential. Here we report on<br />
two main questions. First, we ask if C. capitata is aided during the invasion process by<br />
phenotypic plasticity of thermal tolerance relative to a narrowly-distributed congener C. rosa.<br />
Second, we investigated thermal tolerance and their responses to temperature acclimation<br />
conditions in both species.<br />
Methods: First, we examined rapid cold hardening (RCH) and rapid heat hardening (RHH) in<br />
C. rosa and C. capitata by investigating the effects of sub-lethal low and high temperature<br />
pretreatments on low and high temperature survival respectively. Since RCH was more<br />
pronounced than RHH, we then further examined the time course of RCH. Using a population<br />
extinction model in various thermal habitat scenarios, coupled with microclimate data where<br />
both species cohabit, we then estimated the potential fitness effects that variation in RCH<br />
might afford C. capitata under natural conditions. Second, using a full-factorial design, the<br />
responses of upper and lower critical thermal limits (CTLs) to one week of acclimation at three<br />
mild temperatures (20, 25 and 30 °C) and their interactions with 2 h exposures to these same<br />
temperature conditions were investigated. Lastly, the effects of temperature ramping rates on<br />
CTLs were investigated to better comprehend ecologically-relevant thermal limits to fruit fly<br />
activity.<br />
Results: The range of temperatures inducing RCH were similar for both species (5-10 °C) but<br />
at 5 °C C. capitata developed RCH significantly faster than C. rosa and which also persisted<br />
longer (8 h vs. 0.5 h) than its congener. A simple population survival model, based on the<br />
estimated time-course of RCH showed that time to extinction is greater for C. capitata than for<br />
C. rosa, especially in habitats where temperatures frequently drop below 10°C. Acclimation<br />
experiments showed improvement in heat tolerance with high temperature acclimation and an<br />
increase in low temperature tolerance following acclimation to cooler conditions although in<br />
some cases, significant interaction effects were evident for CTLs between short- and long-term<br />
temperature treatments. Slower heating rates also resulted in higher CTmax while slower<br />
cooling rates elicited lower CTmin.<br />
Conclusions: Variation in RCH responses may translate into significant variation in survival<br />
upon introduction to novel thermal habitats for C. capitata, particularly in cooler and more<br />
thermally variable geographic regions and may contribute to their ongoing invasion success<br />
relative to other, more geographically-constrained Ceratitis species. Acclimation results<br />
suggest that CTLs of adult fruit flies are moderated by temperature variation at both short and<br />
long timescales and may comprise both reversible and irreversible components<br />
Keywords: Acclimation; critical thermal limits; fruit flies; invasive; rapid cold-hardening<br />
51
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FEMALE RECEPTIVITY MODULATION IN ANASTREPHA FRATERCULUS: THE<br />
ROLE OF SPERM AND MALE ACCESSORY GLANDS PRODUCTS.<br />
Abraham, Solana *1,2 ; Willink, Eduardo 1 , Cladera, Jorge L. 3 & Vera, M. Teresa 1,2<br />
1 Sección Zoología, EEAOC, Tucumán, Argentina; Email: solanaabraham@yahoo.com.ar; 2 CONICET, Argentina;<br />
3 Instituto de Genética, INTA Castelar, Buenos Aires, Argentina.<br />
Background: Renewal of receptivity in females may reduce reproductive success of the first<br />
male. It is expected that males develop ways to manipulate female receptivity and avoid mixture<br />
of rival’s sperm. In several fruit flies, at least two mechanisms were examined: sperm effect and<br />
male accessory glands products (AGPs) effect. Here we evaluated the role of sperm and AGPs<br />
in the renewal of A. fraterculus female receptivity and how this was affected by the diet the<br />
male was provided and male origin.<br />
Methods: First, females were mated with males fed either sugar, low quality protein (3:1 sugar:<br />
protein ratio), high quality protein (same ratio) and the standard adult diet (sugar, protein,<br />
vitamins and corn hydrolyze). Wild and laboratory flies were evaluated. After the first<br />
copulation, different sets of females were used to determine female remating rate, refractory<br />
period, and sperm stored. Second, we evaluated the receptivity of females microinjected with<br />
AGPs in two set of trials. In the first, laboratory females were injected with AGPs of laboratory<br />
males fed either with sugar or the standard diet. In the second, laboratory and wild females were<br />
injected with AGPs of laboratory and wild males, all of them fed with standard diet. Female<br />
receptivity was evaluated 48 h after injection and the control were females injected only with<br />
saline. Differences in the frequency of remating females among treatments were compared with<br />
� 2 -tests with Bonferroni’s correction. Refractory period and amount of sperm stored were<br />
analysed with ANOVA coupled with Tuckey’s test.<br />
Results: Wild females mated with sugar males presented a higher remating rate and a shorter<br />
refractory period than those mated with standard diet males. Amount of sperm stored in females<br />
mated with sugar or low quality protein males was significantly lower than that stored in<br />
females that mated with standard diet males. Laboratory females also showed a lower refractory<br />
period when mated with sugar males compared to females mated with protein males irrespective<br />
of the protein quality. However, remating rate and amount of sperm stored was not affected by<br />
the diet. Additionally, females injected with AGPs of laboratory standard diet males had lower<br />
receptivity than females injected with AGPs of laboratory sugar males, which in turn were equal<br />
to control females. Females injected with AGPs from laboratory males had lower receptivity<br />
than females injected with AGPs from wild males (which were equal to the control),<br />
irrespective of female origin.<br />
Conclusions: Male nutritional status and origin affects female renewal of the receptivity. In wild<br />
flies this seems to be related with the sperm stored in the female. However in laboratory flies,<br />
the sperm effect seems to be not the only responsible of modulating these changes. Male<br />
nutritional status and origin also affected female receptivity trough an AGPs effect. The later<br />
may be related to the crowded conditions under laboratory rearing where laboratory males may<br />
need to increase the effectiveness of their AGPs to ensure its paternity.<br />
Keywords: polyandry, remating inhibition, sperm effect, accessory gland effect, South American fruit fly.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ABUNDANCE OF FRUIT FLY SPECIES AND THEIR PARASITOIDS COLLECTED ON<br />
COFFEE CROPS UNDER SHADED AND SUN CONDITIONS IN COLOMBIA.<br />
Giraldo-Jaramillo, Marisol* 1 ; Benavides, Pablo 1 ; Farfán, Fernando 2; Rendon, José Raúl 3 ,<br />
Mejia, Carlos Gonzalo 3 & Montoya, Diego 3<br />
1 Centro Nacional de Investigaciones de Café (Cenicafé), Disciplina de Entomología. Email:<br />
Marisol.Giraldo@cafedecolombia.com; 2 Disciplina de Fitotecnia, 3 Programa de experimentación. Planalto, Km. 4<br />
Antigua vía Chinchiná- Manizales, Caldas, Colombia.<br />
Background: This research is being conducted in order to identify Anastrepha species and their<br />
parasitoids related to two conditions of coffee systems: shaded and sun exposed coffee crops.<br />
Methods: Twenty-four McPhail traps with protein hydrolysate as feeding attractant were placed<br />
randomly on five farms in four departments, Quindio (1), Caldas (2), Risaralda(1) and Valle<br />
del Cauca (1), located at the main coffee crop region in Colombia. Half the traps were disposed<br />
in shaded coffee crops, while the others were set under sun exposed conditions. The shaded<br />
trees species were as follow: avocado Persea americana, guamos Inga spp., guava Psidium<br />
guajava, and Manga Mangifera indica. Sun exposed did not have any other tree species at all.<br />
Fruit fly adults have been weekly collected and registered from the traps up to eight months.<br />
We performed frequency and constancy analysis with this information. Simultaneously, we<br />
collected 30 samples of coffee berries of a pound each, in order to calculate the percentage of<br />
parasitism and to register parasitoid species.<br />
Results: Results showed total captures of 10,112 fruit fly individuals, where 5,512<br />
corresponded to females and 4,600 to males of the Anastrepha genus (Tephritidae: Diptera).<br />
Anastrepha fraterculus was the most frequent species accounting 80% of the total captures,<br />
followed by A. obliqua with 9% frequency and A. striata with 7%. Other five species were<br />
identified with only 4% of the total captures: A. distincta, A. grandis, A. mucronota, A.<br />
manihoti, and A. pallidipennis. While in Valle del Cauca, Quindio and Risaralda the species A.<br />
fraterculus was the dominant species ranging from 97 to 99% frequency, A. obliqua and A.<br />
striata represented 14% of the total captures in Caldas and A. fraterculus 65%. In the whole<br />
experiment, 77% of the fruit fly individuals were captured under shaded conditions. The<br />
parasitism found from the 30 samples evaluated was 18.5%. So far, three genera of parasitoids<br />
were registered only in coffee crops under shaded conditions: Doryctobracon sp., Microcasis<br />
sp. and Utetes sp. (Braconidae: Hymenoptera). Doryctobracon sp. was the most frequent<br />
species.<br />
Conclusions: Anastrepha was the only fruit fly genus collected at the main coffee growing area<br />
of Colombia. A. fraterculus was the most frequent species at both conditions shaded and sun<br />
exposed coffee crops.. Other seven species of fruit flies were identified living on coffee<br />
ecosystems. We found three parasitoids species of fruit flies under shaded coffee crop<br />
conditions which belonged to the Braconidae family.<br />
Keywords Fruit flies, Coffee grown, Anastrepha, Braconidae.<br />
53
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FEMALE ANASTREPHA SUSPENSA (LOEW) RESPONSE TO THE VIBRATION<br />
COMPONENT OF MALE WING-FANNING SIGNALS.<br />
Mankin, Richard W.* & Egan, Mackenzie A.<br />
USDA-ARS Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23 rd Dr, Gainesville, Florida<br />
USA 32608; Email: Richard.Mankin@ars.usda.gov<br />
Background: Anastrepha suspensa is an important pest of fruit crops in the Caribbean region.<br />
Courtship and mating behaviors have been analyzed in many previous studies to develop<br />
control methods. During courtship, males group in leks on leaves of host trees, fan their wings,<br />
and release pheromone to attract females. The wing fanning vibrations are transferred though<br />
their legs to the leaves and nearby stems. We hypothesized that females detect these wingfanning<br />
vibrations, and their behavioral responses may affect the likelihood of mating.<br />
Methods: Wing-fanning vibrations of male A. suspensa on leaves of strawberry guava host<br />
branches were recorded with accelerometers. The vibrations were similar in spectral and<br />
temporal pattern to sounds detected simultaneously by microphones. A 2-min file was created<br />
from a representative, 11-s period of male calling with 1 min continuously repeating the 11-s<br />
call, followed by 1-min silence. The file was played back in an endless loop by a vibration<br />
exciter attached to a mesh cage containing 10 females. The behaviors of the females were<br />
observed remotely at the midpoint of each 1-min calling or silence period using a video camera<br />
during 20-min playbacks. Movement was defined as activities typical of avoidance or<br />
disturbance behavior, including spinning around, walking, wing-fanning, and flying. If the<br />
female was resting or grooming at the calling or silence midpoint, she was scored as quiet. The<br />
total numbers moving or quiet at each midpoint were summed to score each test. Two groups<br />
of females were tested to consider possible effects of prior exposure to males. One group (13<br />
tests) was exposed to pheromone produced by a cage of calling males on the day previous to<br />
testing. The other group (12 tests) was never exposed to males or male pheromone prior to<br />
testing. All tests were conducted during times of day optimal for mating.<br />
Results: Fewer females moved during the periods when calling vibrations were played than<br />
during periods of silence. For females who had never been exposed to male pheromone, a<br />
mean of 12.7 females per 20-min test were moving at the midpoint of the vibration period<br />
compared to 16.8 at the midpoint of the silent period (t = -2.44, df = 12, P = 0.03), i.e., 0.13<br />
movements/fly/min of broadcast compared to 0.17 movements/fly/min of silence. When<br />
females were pre-exposed to males on the day before testing, a mean of 14.4 were moving at<br />
the midpoint of the vibration period compared to 19.25 at the midpoint of the silent period (t =<br />
-2.84, df= 11, P = 0.02) or 0.14 movements/fly/min of broadcast compared to 0.19<br />
movements/fly/min of silence.<br />
Conclusions: The results suggest that female A. suspensa can detect the wing-fanning<br />
vibrations from the male calling song on the host plant, just as they detect the sounds of the<br />
calls, and that the vibrations may reduce the likelihood that the female will move away from<br />
the male who approaches her to mate. The observed effects of vibrations are similar in many<br />
respects to the effects of wing-fanning sounds on females, and the two signals may have<br />
synergistic effects. Could disruption of male vibrations be a potential control method?
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SPATIAL AND DIURNAL PATTERN OF PROTEIN FORAGING BY BACTROCERA<br />
TRYONI (FROGGATT) ON A HOST PLANT.<br />
Balagawi, Solomon 1,2 *; Jackson, Kevin 2,3 ; Hamacek, Edward 3 & Clarke, Anthony R. 1,2<br />
1 Discipline of Biogeosciences, Queensland University of Technology, G.P.O. Box 2434, Brisbane 4001,<br />
Queensland, Australia. Email: solomon.balagawi@qut.edu.au; 2 Cooperative Research Centre for National Plant<br />
Biosecurity, Canberra, Australia; 3 Horticulture and Forestry Science, Agri-Science Queensland, Department of<br />
Employment Economic Development & Innovation, Brisbane, Australia.<br />
Background: Queensland fruit fly, Bactrocera tryoni, is the major pest fruit fly in Australia.<br />
Protein bait sprays, where insecticides are mixed with spot applications of a protein based food<br />
lure, are one of the sustainable pre-harvest fruit fly management strategies used in Australia.<br />
Although protein bait sprays do manage fruit fly infestation in the field, there is little science<br />
underpinning this technique and so improving its efficacy is difficult. Lacking information<br />
includes where and when to apply protein bait in order to best target foraging B. tryoni. As part<br />
of new work in this area, we investigated the effect of height of protein on tree and host plant<br />
fruiting status on the spatial and temporal protein foraging patterns of B. tryoni.<br />
Methods: The work was conducted in the field using nectarine and guava plants and wild B.<br />
tryoni at Redland Bay, Queensland, Australia. Spot sprays of protein bait were applied to the<br />
foliage of randomly selected fruiting and non-fruiting trees. Each tree received protein bait spot<br />
sprays on the lower and higher foliage at 0530hrs. The number, sex and species of flies that fed<br />
on each protein spot were recorded hourly from 0600hrs through to 1800hrs.<br />
Results: For nectarines, there was a significant difference in the number of B. tryoni feeding on<br />
protein bait placed at different locations within the tree (ANOVA, F = 8.898, p = 0.001). More<br />
flies fed on protein placed on higher foliage relative to lower, irrespective of the fruiting status<br />
of the nectarine trees. A significant difference was also observed in the diurnal protein feeding<br />
pattern of B. tryoni (ANOVA, F = 2.164, p = 0.024), with more flies feeding at 1600hrs.<br />
Results for guava are still being collected and will be presented at the meeting.<br />
Conclusions: We conclude that B. tryoni effectively forages for protein at heights higher than<br />
1.3m from ground, indicating greater efficacy of protein bait when applied at foliage higher in<br />
the canopy. Bactrocera tryoni actively forages for protein throughout the day, with a highest<br />
feeding peak at 1600hrs. The lack of significant difference in the spatial protein foraging<br />
pattern between fruiting and non-fruiting nectarine trees may be a real result, or may have<br />
resulted from the fruiting tree being very close (within 1 – 2 metres) of the non-fruiting tree.<br />
This hypothesis is being tested in the guava trial.<br />
Keywords: protein bait, resource use<br />
55
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CLASSICAL AND MOLECULAR STUDY OF GUT BACTERIAL COMMUNITY STRUC-<br />
TURE IN ALIMENTARY TRACT OF BACTROCERA CUCURBITAE (COQUILLETT).<br />
Thaochan, Narit* 1 ; Chinajariyawong, Anuchit 2 & Drew, Rai 3<br />
1 Department of Pest Management, Faculty of Natural Resources, Prince of Songkla University, Hat Yai,<br />
Songkhla, 90110, Thailand. Email: narit_taochan@yahoo.com. 2 National Biological Control Research Center<br />
(NBCRC), Central Regional Center, Kamphaeng Saen, Nakhon Pathom 73140, Thailand. 3 International Centre for<br />
the Management of Pest Fruit Flies, Australian School of Environmental Studies, Griffith University, Nathan<br />
Campus, Nathan, Queensland, 4111, Australia.<br />
Background: Bactrocera cucurbitae (Diptera: Tephritidae) is recognized as an important insect<br />
pest species in many cucurbitaceous plants. The bacteria in fruit flies alimentary tract have<br />
been known in advantages in their biology of the insect hosts. The classical techniques of gut<br />
bacterial study in fruit fly were known with easy culturable gut bacteria but unculturable and<br />
not-yet culture bacteria could not be detected. Recently, molecular techniques can reveal real<br />
time of gut bacterial community structure in insect alimentary tract. In this research, we employed<br />
classical and molecular cloning techniques to study gut bacterial community structure<br />
in B. cucurbitae gathered from wild.<br />
Methods: Both males and females of B. cucurbitae were hand-collected from their natural host<br />
plants in Nakhon Si Thammarat province, Thailand. Crop and midgut of the flies were dissected<br />
with aseptic techniques. The classical technique was the isolation of the gut bacteria on<br />
culture media and identified with API 20 E test kit. For the molecular technique, DNAs were<br />
extracted from their alimentary tracts. Genomic DNAs were amplified with universal primer of<br />
16S rRNA gene for bacteria. PCR products were transfer to competent cell and selecting by<br />
cloning techniques. The clone containing interested gene were sequenced and analyzed on<br />
RDP II. The community diversity and community similarity were analyzed with Shannon-<br />
Weaver index, rarefaction analysis, Morisita’s index, multidimensional scaling analysis (MDS)<br />
and phylogenetic tree analysis.<br />
Results: Classical technique revealed only one bacterial family Enterobacteriaceae that belonged<br />
to Gramma-proteobacteria in this fruit fly species. In molecular study, five bacterial<br />
taxa, Alpha-, Beta-, Gramma-proteobacteria, Actinobacteria and Firmicutes, were found. The<br />
former three bacterial taxa were classified in phylum Proteobacteria. Interestingly, Wolbachia<br />
sp., known as bacterial arthropod pathogen and unculturable bacterium, was detected. Female<br />
midgut showed the highest values of gut bacterial community diversity. The gut bacterial<br />
community similarity competently distinguished the differences of their gut bacteria harbored<br />
between crop and midgut.<br />
Conclusions: Molecular cloning techniques in this study have exposed appreciation of the<br />
ecology of gut bacterial community structures in B. cucurbitae alimentary canal. More bacterial<br />
species, including unculturable bacteria and also rare species were detected. Grammaproteobacteria<br />
was the predominant bacterial taxon in this fruit fly species.<br />
Keywords: Gut bacterial community structure, Alimentary tract, Molecular cloning technique, Bactrocera cucurbitae,<br />
Gramma-proteobacteria
Session 2<br />
Morphology & Taxonomy
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SIMULTANEOUS DETECTION AND IDENTIFICATION OF TEN DIFFERENT SPECIES<br />
OF TEPHRITID FRUIT FLIES USING OLIGONUCLEOTIDE MICROARRAYS.<br />
Chen, Yen-Hou 1 , Liu, Lu-Yan 2 , Tsai, Wei-Huang 3 & Lu, Kuang-Hui 1 *<br />
1 Department of Entomology, National Chung Hsing University, No. 250, Kuo-Kuang Rd., Taichung 40227,<br />
Taiwan. Email: khlu@dragon.nchu.edu.tw; 2 Graduate Institute of Plant Science, National Pintung University of<br />
Science and Technology, No. 1, Shuehfu Road, Neipu, Pingtung 91201, Taiwan. 3 Division of Plant Protection,<br />
Bureau of Animal and Plant Health Inspection and Quarantine, Council of Agriculture, Executive Yang, 9Dl., No.<br />
51, Sec. 2, Chungching S. Rd., Taipei 10075, Taiwan<br />
Background: Tephritid fruit flies are the most destructive insect pests of horticultural crops<br />
worldwide. To prevent the invasion of exotic tephritids, many countries set strict regulation on<br />
the quarantine inspection for imported fruits and vegetables. A method for rapid identification<br />
of the species of intercepted pests, which are most at their larval or even egg stages, are<br />
required to enhance the efficiency and quality of quarantine inspection and service. Here, we<br />
report that we have developed a microarray to meet the demand for rapid identification of<br />
insect species.<br />
Methods: Ten species of tephretid fruit flies, i.e., Bactrocera dorsalis, B. cucurbitae, B. tau, B.<br />
oleae, B. papayae, B. tryoni, B. carambolae, Ceratitis capitata, Anastrepha suspensa,<br />
Rhagoletis pomonella, were selected for the experiment. The mitochondrial genes, including<br />
cytochrome oxidase I and II (COI and COII) and NADH dehydrogenase subunit 4 and 5 (ND4<br />
and ND5), were targeted for species-specific probe screening. The genes were amplified by<br />
PCR using appropriate universal primers; the PCR products were cloned and sequenced; and<br />
the DNA sequences of each gene of all species were aligned for designing oligonucleotide<br />
probes. The probes were spotted on the microarray polymer substrate and immobilized by a<br />
UV cross-linker. The hybridization and colorimetric reactions between each target DNA and<br />
probe were carried out with DR. Chip DIY TM Kit (DR. Chip Biotech, Miao-Li, Taiwan).<br />
Results: (1) a multiplex PCR with four pairs of primers, i.e., COI, COII, ND4 and ND5, were<br />
developed, and it was able to simultaneously amplify all four target DNA fragments of all<br />
tested flies. (2) Multiplex probes of different oligonucleotide sequences targeting the four<br />
genes were obtained, and they were capable of discriminating one species unambiguously from<br />
others except between B. dorsalis and B. papayae.<br />
Conclusions: We have developed an inexpensive microarray that enables to rapidly and<br />
simultaneously indentify at least ten species of major tephretid pests within 5 hours.<br />
Keywords: tephritid fruit fly, microarray, cytochrome oxidase, NADH dehydrogenase, plant quarantine<br />
59
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
A HOLISTIC APPROACH TO DETERMINING SPECIES LIMITS BETWEEN<br />
MORPHOLOGICALLY SIMILAR PEST SPECIES OF THE BACTROCERA DORSALIS<br />
SPECIES COMPLEX: INITIAL RESULTS USING GEOMETRIC MORPHOMETRICS.<br />
Schütze, Mark K. 1,2 *; Jessup, Andrew J. 3 & Clarke Anthony R. 1,2<br />
1 Discipline of Biogeosciences, Queensland University of Technology, G.P.O. Box 2434, Brisbane 4000,<br />
Queensland, Australia. Email: m.schutze@qut.edu.au. 2 Cooperative Research Centre for National Plant<br />
Biosecurity, Canberra, Australia. 3 Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear<br />
Techniques in Food and Agriculture, International Atomic Energy Agency, Vienna, Austria.<br />
Background Particular pest species of the Bactrocera dorsalis species-complex are<br />
taxonomically indistinguishable with high intra-specific morphological variation. The most<br />
problematic species are B. dorsalis sensu stricto, B. papayae, B. carambolae, and B.<br />
philippinensis. Doubt remains over the taxonomic validity and species limits of these taxa. We<br />
aim to test this hypothesis by examining different biological aspects of these species.<br />
Methods The broad study incorporates multiple lines of evidence including cross-mating trials<br />
and comparing physiological characteristics among these species. This information will be<br />
coupled with genetic, morphological, and morphometric data. Work completed includes a<br />
preliminary geometric morphometric analysis of collection material to resolve fine scale<br />
differences among these taxa as currently described. For the geometric morphometric study, 15<br />
landmarks were applied to wings of 245 collection specimens coming from three populations<br />
of each of the above mentioned species, plus one outgroup, B. tryoni. Generalized Procrustes<br />
analysis was then performed, from which canonical variates analyses (CVA) were undertaken<br />
to determine if defined species could be discriminated. Hypothetical ‘unknown’ samples were<br />
also included to assess if this technique could assign unknown individuals to species based on<br />
shape data.<br />
Results The geometric morphometric study revealed that intraspecific wing size did not vary<br />
significantly between populations within species, but significant interspecific size comparisons<br />
did occur. Of the ingroup species, B. papayae had the smallest wings (average centroid size =<br />
6.002 ± 0.061 s.e.), while B. dorsalis s.s. had the largest (6.349 ± 0.066). CVA clustered<br />
together populations within a single species, but discriminated between species. The single<br />
exception was an intraspecific difference between one population of B. carambolae and the<br />
two other conspecific populations. This statistically unusual population was subsequently<br />
deemed an original misidentification and likely represents an undescribed species. Individuals<br />
were correctly reassigned to their respective species on average 93.27% of the time; the<br />
assignment of 15 ‘unknowns’ was 80% successful.<br />
Conclusions We conclude that geometric morphometric data is applicable to quantifying shape<br />
variation for these species. With the generation of further biological data, we anticipate our<br />
understanding of species limits for this group will significantly improve, as will our ability to<br />
develop diagnostic markers.<br />
Keywords: geometric morphometrics; species limits; cryptic species
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE ANASTREPHA FRATERCULUS SPECIES COMPLEX: RECOGNITION OF THREE<br />
MORPHOTYPES FROM ANDEAN COUNTRIES USING THE MULTIVARIATE<br />
APPROACH.<br />
Hernández-Ortiz, Vicente* 1 ; Selivon, Denise 2 ; Bartolucci, Andrea 3 ; Morales-Valles,<br />
Pedro 4 & Frías, Daniel 5<br />
1 Instituto de Ecología A.C. (INECOL), Red de Interacciones Multitróficas, Apartado Postal 63. Xalapa, Veracruz<br />
91070, México. Email: vicente.hernandez@inecol.edu.mx; 2 Universidade de Sao Paulo (USP), Departamento de<br />
Biología, Instituto de Biociencias, Sao Paulo, Brazil; 3 Instituto de Sanidad y Calidad Alimentaria de Mendoza<br />
(ISCAMEN), Programa de Erradicación de Mosca del Mediterraneo, Ciudad Mendoza, Argentina; 4 Instituto de<br />
Investigaciones Agrícolas (INIA-CENIAP), Protección Vegetal, Maracay, Aragua, Venezuela; 5 Universidad<br />
Metropolitana de Ciencias de la Educación (UMCE) Instituto de Entomología, Santiago, Chile.<br />
Background: One of the most important fruit fly species in continental America is the ASouth<br />
American fruit fly@ Anastrepha fraterculus (Wiedemann), recorded from southern Texas<br />
(USA) through Mexico, Central America, and South America. This nominal species is<br />
currently recognized as a cryptic species complex, based on karyotypic differences, isozymic<br />
analyses, mitochondrial DNA sequences, or host use patterns, among others. Despite all this<br />
evidence, to date there are no taxonomic elements to ascertain )how many species there are in<br />
this complex?, or )how they can be distinguished from each other?. A recent morphometric<br />
analysis from several Mexican populations of the nominal A. fraterculus showed that they can<br />
be distinguished from other samples belonging to South America. In this study we assessed the<br />
morphometric variability using the same approach in ten natural populations collected from<br />
four South American countries.<br />
Methods: We analyzed 10 population samples from Colombia (2), Venezuela (5), Ecuador (1)<br />
and Perú (2) using 15 specimens by sample. On the basis of structures as the aculeus,<br />
mesonotum and wing, 19 morphometric traits were assessed for each specimen. Discriminant<br />
function analysis (DFA) were applied to the full data set, grouping the specimens by locality of<br />
origin. The functions were tested by canonical correlation analysis to assess the significance of<br />
the discriminatory power of the model and variables involved in the segregation of the groups.<br />
Results: Our results showed that there are clear differences between populations, even though<br />
all of them become from the biogeographical sub-region of Northwestern South America.<br />
Samples from the highlands of Colombia and Venezuela were very similar to each; a second<br />
group was differentiated from a single population of the Venezuelan lowlands; while samples<br />
from the lowlands of Peru and Ecuador were grouped separately.<br />
Conclusions: In the Andean countries the AF complex would be represented by three natural<br />
groups here called morphotypes: the Venezuelan morphotype, which occurs at lowlands from<br />
that country; the Andean morphotype assembled by highland populations from Venezuela and<br />
Colombia; and the Peruvian morphotype recognized by populations that occur along the Pacific<br />
coastal slopes from Peru and Ecuador.<br />
Keywords: South American Fruit Fly, cryptic species, morphometry, morphotypes<br />
61
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DIVERGENCE OF FRUIT FLIES IN THREE SPECIES GROUPS OF THE GENUS<br />
RHAGOLETIS IN MÉXICO.<br />
Rull, Juan 1* ; Aluja, Martín 1 ; Feder, Jefferey. L 2 ; Guillén, Larissa 1 & Tadeo, Eduardo 3<br />
1<br />
Red de Manejo Biorracional de Plagas Instituto de Ecología, A.C., Xalapa, Veracruz, México.<br />
juan.rull@inecol.edu.mx; 2 Department of Biological Sciences, Galvin Life Science Bld., University of Notre<br />
Dame, Notre Dame IN, 46530, USA; 3 Instituto de Neuro Etología, Universidad Veracruzana, Xalapa, Veracruz,<br />
México.<br />
Background: Fruit flies in the genus Rhagoletis spearheaded the long and heated debate about<br />
the possibility and later the frequency of occurrence of sympatric speciation via host shifting<br />
for more than forty years. Once sympatric speciation was accepted as a common route to<br />
divergence in phytophagous insects, careful study of Mexican populations of the apple maggot<br />
fly Rhagoletis pomonella Walsh revealed that differentiation in the pomonella group had been<br />
in fact possible due to a combination of periods of allopatric divergence, gene flow and<br />
introgression and recent sympatric speciation via host shifting events.<br />
Methods: Such findings led us to undertake detailed philogeographic studies of the genus in<br />
México. We first studied the distribution, divergence and evolution of reproductive isolation<br />
among hawthorn infesting populations of Rhagoletis in Mexico and the U.S. We are now<br />
conducting similar studies on black cherry infesting populations in the cingulata species group,<br />
and walnut infesting populations in the suavis species group.<br />
Results: Despite the fact that flies in the three species groups shared a similar climatic history<br />
of range contraction and expansion, divergence in the three groups appears to be quite<br />
different, with flies in the pomonella group exhibiting what is now known as speciation mode<br />
plurality, flies in the suavis group undergoing rapid morphological variation without<br />
reproductive isolation and introgression between some species, and flies in the cingulata group<br />
exhibiting classic vicariant speciation.<br />
Conclusions: We discuss hypotheses on the outcome of these studies.<br />
Keywords: Rhagoletis, philogeography, divergence, speciation
Session 3<br />
Genetics & Evolution
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
NEW TOOLS FOR THE DEVELOPMENT AND STABILIZATION OF TRANSGENIC LINES<br />
OF CERATITIS CAPITATA (WIEDEMANN).<br />
Scolari, Francesca 1 ; Schetelig, Marc F. 3 ; Bertin, Sabrina 1 ; Handler, Alfred M. 3 ; Kittelmann, Sebastian 2 ;<br />
Malacrida, Anna R. 1* ; Gasperi, Giuliano 1 & Wimmer, Ernst A. 2<br />
1 Dipartimento di Biologia Animale, Università di Pavia, Via Ferrata 1, 27100 Pavia, Italy. 2 Department of<br />
Developmental Biology, Göttingen Center for Molecular Biosciences, Johann-Friedrich-Blumenbach-Institute of<br />
Zoology and Anthropology, Georg-August-University Göttingen, GZMB, Justus-von-Liebig-Weg 11, 37077<br />
Göttingen, Germany. 3 United States Department of Agriculture, Agricultural Research Center, Center for Medical,<br />
Agricultural and Veterinary Entomology, 1700 Southwest 23 rd Drive, Gainesville, FL 32608.<br />
Background. The Mediterranean fruit fly (medfly) (Wiedemann; Diptera: Tephritidae),<br />
Ceratitis capitata, for which a successful SIT package has been developed and validated, was<br />
the first non-drosophilid insect to be transformed, opening the way for the genetic<br />
transformation of many other pest insects that are targets of SIT programmes. The efficacy and<br />
cost effectiveness of the SIT both at the mass-rearing, release and monitoring stages may be<br />
improved by using different medfly strains currently available, which provide i) genetic<br />
marking for the identification of transformed insects; ii) male-specific fluorescent sorting; iii)<br />
sexing for male-only strains; iv) reproductive sterility through embryonic lethality.<br />
Methods. For the efficient application of SIT programmes, effective monitoring to assess the<br />
number and mating success of the released flies is essential. With this aim, we developed a<br />
powerful sperm-specific marking systems based on the spermatogenesis-specific C. capitata<br />
β2-tubulin (Ccβ2t) promoter. Molecular characterization and preliminary laboratory<br />
competitiveness assays identified a target line which was then modified by the site-specific<br />
integration system from phage phiC31. Donor plasmids containing an attB site, with additional<br />
markers, and transposon ends were integrated into attP sites randomly integrated within<br />
transposon-based vector by phiC31 integrase-mediated recombination. Finally, transposaseencoding<br />
‘jumpstarter’ strains were created and mated to transgenic strains to produce postintegrational<br />
excision of transposon ends.<br />
Results. Several lines were generated in which the reporter genes tGFP or DsRedEx are<br />
expressed in a stable, persistent and sex-specific manner during spermatogenesis. Fluorescent<br />
sperm can be isolated from testes or spermathecae. The marking does not result in fitness costs<br />
in preliminary laboratory assays. Particularly, males of one sperm-marked line showed no<br />
significant reduction in their overall fitness, transmitting their genes to the next generation in a<br />
competitive way. Site-specific integration experiments performed on this line resulted in the<br />
post-integrational excision of a transposon end, which left stably integrated transgene<br />
insertions that could not be further remobilized.<br />
Conclusions. The development of a sperm marking system in the medfly and the subsequent<br />
stabilization of the transgenic lines represent a powerful tool since i) the use of such harmless<br />
transgenic markers are an ideal for the transfer of insect transgenesis technology from the<br />
laboratory to field applications; ii) effective and easily recognizable sperm marking will make<br />
novel studies possible on medfly reproductive biology which will in turn help to further<br />
improve environmentally safe, biological pest management approaches; iii) the newly<br />
developed integration and stabilization system will allow the combination of several transgeneencoded<br />
advantageous traits at evaluated genomic positions to generate optimized strains for<br />
pest control that minimize environmental concerns.<br />
Keywords: sperm marking, insect pest management, phiC31 integrase, transgene stability.<br />
65
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GENETIC ENGINEERING OF THE OLIVE FRUIT FLY, BACTROCERA OLEAE, FOR USE<br />
IN THE STERILE INSECT TECHNIQUE (SIT).<br />
Ant, Thomas H. 1, 2 ; Koukidou, Martha* 1 ; Morgan, Sian A. 1 1, 2<br />
& Alphey, Luke<br />
1 Oxitec Ltd, 71 Milton Park, Abingdon, OX14 4RX, Oxfordshire, UK; Email: martha.koukidou@oxitec.com;<br />
2 University of Oxford, Department of Zoology, Oxford, UK.<br />
Background: The olive fruit fly, Bactrocera oleae (Diptera: Tephritidae), is the most<br />
destructive pest of olive fruit, causing considerable crop damage in the Mediterranean region<br />
and in California. Despite significant spending on insecticides the olive fly causes a reduction<br />
in crop yield and quality. Furthermore, some of the insecticides used are being phased out and<br />
olive fly is developing resistance to insecticides (even to relatively new insecticides such as<br />
Spinosad). The sterile insect technique (SIT) is a species-specific and environmentally nonpolluting<br />
method of pest control that involves the mass release of sterile insects that has been<br />
very effective against other fruit flies. Previous SIT attempts using irradiated mixed-sex insects<br />
achieved only limited success at suppressing populations of B. oleae. The released sterile males<br />
mated with the released sterile females, instead of dispersing and seeking the wild-type<br />
females. It was believed at the time that sterile flies preferred to mate earlier in the day, leading<br />
to partial reproductive isolation from with wild population. A genetic sexing system to allow<br />
male only release is therefore seen as essential for olive fly SIT; while also providing the<br />
several benefits provided by current Medfly genetic sexing strains<br />
Method: We used genetic transformation methods to develop engineered strains of olive fly<br />
(‘RIDL strains’) and used modifications of standard Medfly protocols to test strain<br />
performance.<br />
Results: Oxitec has developed genetic sexing strains of olive fly based on a conditional positive<br />
feedback design already successfully applied in two other tephritid species; the Mediterranean<br />
fruit fly, Ceratitis capitata, and the Mexican fruit fly, Anastrepha ludens. The strains are 100%<br />
effective in separating sexes and also incorporate a fluorescent marker allowing easy<br />
monitoring in the field. Furthermore, since females die without a dietary supplement<br />
(tetracycline) that is not available in the wild, significant bio-containment is also inherent in<br />
this design of sexing strain. Oxitec is currently conducting laboratory and greenhouse mating<br />
performance tests with these sexing strains to further optimize them for SIT.<br />
Conclusions: Our current RIDL strains of olive fly appear to give excellent sex-separation<br />
(‘genetic sexing’) without obvious negative effects on other performance parameters; our latest<br />
data on performance will be presented. The development of these strains greatly improves the<br />
prospects for the use of SIT to control field populations of olive fly.<br />
Keywords: olive fly (Bactrocera oleae), RIDL ® , SIT, genetic sexing
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GENOMIC SEQUENCING OF THE MEDITERRANEAN FRUIT FLY, CERATITIS<br />
CAPITATA.<br />
Handler, Alfred* 1 ; Gasperi, Giuliano 2 ; Richards, Stephen 3 ; Scherer, Steven 3 ; Gomulski,<br />
Ludvik 2 ; Schetelig, Marc 1 & Malacrida, Anna 2<br />
1 Center for Medical, Agricultural, and Veterinary Entomology, USDA, ARS, 1700 SW 23 rd Drive, Gainesville,<br />
Florida 32608 USA; Email: ahandler@ufl.edu; 2 Department of Animal Biology, University of Pavia, Via Ferrata<br />
1, Pavia 27100, Italy; 3 Baylor College of Medicine Human Genome Sequencing Center, Houston, TX 77030<br />
USA.<br />
Background: A joint project between the Baylor College of Medicine Human Genome<br />
Sequencing Center, the U. S. Department of Agriculture, and the University of Pavia was<br />
initiated in 2008 to sequence the whole genome of the Mediterranean fruit fly, Ceratitis<br />
capitata.<br />
Methods: The source material for the project was embryonic genomic DNA extracted from the<br />
medfly Ispra strain maintained as an inbred colony for more than 20 years at the University of<br />
Pavia, though this is considered to be polymorphic. Paired-end libraries were created for the<br />
454 Titanium sequencing platform.<br />
Results & conclusions: Pyrosequencing at 454 Titanium platform of paired-end libraries has<br />
produced 5,179 million bases of genomic DNA sequence after 13 fragment runs. This<br />
represents approximately 10x genomic sequence coverage with an average read length of 348<br />
bp. An initial assembly of the fragment data using 454 Newbler v.2.3 (pre-release-9/14/2009)<br />
software produced 215,255 sequence contigs longer than 500 bp. The average contig size was<br />
1,852 bp and the N50 was 2,709 bp, with the largest contig size of 70,001 bp. The current<br />
contig size data is sub-optimal, and is likely due to high repetitive DNA content and sequence<br />
polymorphism in the sample, negatively affecting the assembly. To improve the assembly<br />
product, new 3 kb insert paired-end libraries for Illumina sequencing have been created to<br />
order and orient the available contigs within the assembly, as well as to add data using a<br />
different sequencing technology. Sex-specific and embryonic RNAseq transcriptome<br />
sequencing is also anticipated. Upon completion of Illumina sequencing (expected in the<br />
summer/autumn of 2010) additional assemblies will be performed, and based on these results<br />
an interim assembly product will be released to public databases.<br />
Keywords: Whole genome sequencing, Medfly, 454 Titanium sequencing, Illumina sequencing<br />
67
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
APPLICATIONS OF GENOME SEQUENCING TO IDENTIFICATION, DETECTION, AND<br />
CONTROL OF BACTROCERA DORSALIS (HENDEL) AND OTHER TEPHRITID PESTS.<br />
Geib, Scott* & Jang, Eric<br />
U.S. Pacific Basin Agricultural Research Center, USDA-ARS, P.O. Box 4459, Hilo, HI 96720; Email:<br />
scott.geib@ars.udsa.gov.<br />
Background: Recent advances in DNA sequencing technologies have increased the feasibility<br />
of performing large scale sequencing experiments in non-model organisms. Next-generation<br />
sequencing (NGS) technologies have allowed for whole genome shotgun (WGS) sequencing<br />
and assembly experiments to be performed in a matter of months for thousands of dollars<br />
compared to the years and millions that it previously took using traditional methods. The<br />
Bactrocera dorsalis genome project is one of the first projects to utilized only NGS<br />
technologies to produce a draft genome assembly of a non-model organism.<br />
Methods: The sequencing of the oriental fruit fly genome was performed by using a<br />
combination of shotgun and paired-end (8 and 20 kb) sequencing with 454 pyrosequencing<br />
utilizing Titanium based chemistry. Approximately 8 Gb of sequence was obtained. A draft<br />
assembly of the genome, utilizing de novo assembly strategies was created using both the<br />
Newbler and the Celera assemblers. From this draft assembly, gene identification and<br />
annotation is currently taking place, utilizing computational pipelines that identify coding<br />
regions, compare the sequence to previously identified genes, and assign function annotation<br />
based on gene ontology. This draft database will likely contain the majority of the genes<br />
present in the oriental fruit fly genome, and can then serve as a database for functional<br />
genomics and proteomics as well as be used as a basis for developing molecular tools for use in<br />
other research projects.<br />
Results: Pyrosequencing on a 454 sequencer using Titanium chemistry was able to produce a<br />
de-novo assembly at approximately 15x coverage. A draft genome of approximately 430 Mb in<br />
size was created. Currently, annotation and analysis of this dataset is ongoing. The utility of<br />
this dataset to the management of this and other Tephritid pests is broad, ranging from<br />
development of genetic markers, such as SNPs and microsatellites that can be utilized for<br />
molecular diagnostics, to functional genomics tools that can be applied to physiological<br />
questions, targeting insect resistance, olfaction, digestion, or other areas.<br />
Conclusions: The integration of a genomic perspective into other research areas opens the door<br />
for many new approaches to asking fundamental questions in invasive species research.<br />
Keywords: Oriental Fruit Fly, genome sequencing, functional genomics, genetic markers
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
HOST PLANT TOXICITY, STENOPHAGY AND EVOLUTIONARY RADIATION IN THE<br />
GENUS CERATITIS (DIPTERA, TEPHRITIDAE).<br />
Erbout, Nathalie *1 ; Lens, Luc 1 ; Virgilio, Massimiliano 2-3 & De Meyer, Marc 2<br />
1 Ghent University, Department of Biology, Terrestrial Ecology Unit, Ledeganckstraat 35, 9000 Ghent, Belgium.<br />
Email: Nathalie.Erbout@ugent.be; 2 Royal Museum for Central Africa (RMCA), Entomology Section, Tervuren,<br />
Belgium. 3Royal Belgian Institute of Natural Sciences, Invertebrates Section, Brussels, Belgium.<br />
Background: It is widely acknowledged that the evolution of insect-host relationships has been<br />
largely influenced by the chemistry of insect host plants. However, whether and to what extent<br />
secondary plant metabolites triggered the evolution of host plant specialization in<br />
phytophagous insects is still much debated. We here describe the phylogeny of host plant<br />
specialization within the genus Ceratitis and unravel the link with host plant toxicity as a<br />
possible mechanism that underlies the evolutionary radiation of stenophagous clades within<br />
this genus.<br />
Methods: Using molecular data from three protein encoding genes and 49 species (98<br />
specimens), we reconstructed the phylogeny of the genus Ceratitis and investigated the<br />
evolution of host plant specialization along the different recognized clades. We used HPLC<br />
analysis to compare the concentration of two toxic secondary metabolites (alkaloids) in host<br />
plants of stenophagous and polyphagous species. In an experimental set-up, we tested if, and to<br />
what extent, development and fitness of a polyphagous fruit fly is adversely affected by host<br />
plant toxicity by comparing rates of development, survival and reproduction on four media that<br />
differ in alkaloid concentration.<br />
Results: Bayesian tree reconstructions supported previously proposed monophyletic lineages.<br />
Reconstruction of ancestral character states for host plant relationships suggested that<br />
stenophagy evolved repeatedly and independently within the genus Ceratitis. HPLC analysis<br />
proved an overall significantly higher concentration of toxic alkaloids in fruits of the<br />
stenophagous host plants, even though their concentration gradually decreased during ripening<br />
process. Despite reduced pupal and adult sizes, polyphagous larvae developing under low<br />
alkaloid concentrations successfully developed to the adult stage, probably as a result of<br />
accelerated pupation and ensuing restricted exposure to the toxic environment. High alkaloid<br />
concentrations, however, impaired their development process and prevented subsequent<br />
reproduction.<br />
Conclusions: The proposed phylogeny shows a number of biological clades including<br />
stenophagous species which share host genera with genus-specific toxic secondary metabolites.<br />
The adverse effects of host plant toxicity on larval development in polyphagous fruit flies<br />
presented here indicate that these high concentrations pose a significant constraint on host use<br />
in polyphagous Ceratitis species. We conclude that the observed phylogenetic patterns for<br />
monophyletic stenophagous clusters are the result of an evolutionary process of ecological<br />
specialization to toxic hosts.<br />
Keywords: Ceratitis, phylogeny, host-plant toxicity, stenophagy, evolutionary radiation<br />
69
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
MACROGEOGRAPHIC POPULATION STRUCTURING IN THE COSMOPOLITAN<br />
AGRICULTURAL PEST BACTROCERA CUCURBITAE (DIPTERA: TEPHRITIDAE).<br />
Virgilio, Massimiliano 1,2* ; Delatte, Hélène 3 ; Backeljau, Thierry 2,4 & De Meyer, Marc 1<br />
1 Royal Museum for Central Africa, Leuvensesteenweg 13, B-3080 Tervuren (Belgium). Email:<br />
massimiliano.virgilio@africamuseum.be; 2 Royal Belgian Institute of Natural Sciences, Vautierstraat 29, B-1000<br />
Brussels (Belgium); 3 UMR C53 PVBMT CIRAD-Université de la Réunion, CIRAD Pôle de Protection des<br />
Plantes, 7 chemin de l’IRAT, Ligne Paradis, 97410 Saint Pierre, La Réunion (France) ; 4 Department of Biology,<br />
University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp (Belgium).<br />
Background: The melon fly Bactrocera cucurbitae (Coquillett) is a major, highly polyphagous,<br />
agricultural pest abundant throughout the African continent, the islands of Indian Ocean, Asia,<br />
New Guinea, the Mariana Islands and Hawaii. This species attacks more than 125 plants<br />
including commercial crops such as pumpkin, cantaloupe, watermelon, squash, gourd,<br />
cucumber, tomato, eggplant and bean as well as soft fruits such as mango, orange, papaya and<br />
peach. Although the economic importance of B. cucurbitae is well documented, its large-scale<br />
(i.e. inter-regional) patterns of genetic structuring are poorly known. Hence, the current<br />
quarantine methods and management plans still rely on general assumptions concerning its<br />
intraspecific variation. The objectives of this work were to 1) characterize the large-scale<br />
population structure of Bactrocera cucurbitae, 2) identify its geographic origin and 3) infer the<br />
dynamics of its range expansion.<br />
Methods: Individuals of B. cucurbitae were collected from 25 worldwide-distributed localities<br />
(n=570) and genotyped at 13 microsatellite loci specifically developed for this species. Genetic<br />
discontinuities among geographical regions and levels of population admixture were quantified<br />
through Bayesian clustering procedures<br />
Results: Five main groups of populations were identified. These corresponded to populations<br />
from 1) the African continent, 2) La Réunion, 3) Central Asia, 4) East Asia and 5) Hawaii. The<br />
proportions of inter-regional assignments and the higher values of genetic diversity in<br />
populations from Pakistan, India and Bangladesh suggest that B. cucurbitae originated in<br />
Central Asia and expanded its range to East Asia and Hawaii on one hand and to Africa and the<br />
islands of the Indian Ocean on the other. A number of outliers (10-19 specimens according to<br />
different clustering algorithms) show high levels of admixture (Q>0.70) with populations from<br />
different regions and reveal complex patterns of inter-regional gene flow.<br />
Conclusions: Anthropogenic transport is the most plausible promoter of the large-scale<br />
dispersal of B. cucurbitae. The recent African invasions of B. cucurbitae were mainly due to<br />
the expansion of local populations while, hitherto, the introduction of individuals from<br />
geographically distant regions did not have a relevant role. The dynamics of the recent<br />
invasions, as well as the possible corridors to gene flow created by contemporary<br />
anthropogenic activities could provide a useful background to better evaluate invasion risks<br />
and establish priorities for the management of this cosmopolitan agricultural pest.<br />
Keywords: Tephritidae, agricultural pests, biological invasions, population structure, microsatellites.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GENETICAL CHANGES IN BACTROCERA TRYONI DURING LABORATORY<br />
ADAPTATION, AND THE USE OF FOUR-WAY HYBRIDS TO REDUCE INBREEDING<br />
EFFECTS<br />
Gilchrist, Stuart; Sved, John* & Meats, Alan<br />
Fruit Fly Research Laboratory, School of Biological, Ecological and Earth Sciences, The University of New South<br />
Wales, Australia; Email: john.sved@sydney.edu.au<br />
Background: Domestication of wild strains for use in factories appears to be a problematic<br />
process in Queensland fruit fly (Bactrocera tryoni), as in most tephritids. Early generations, in<br />
particular, are subject to large fluctuations apparently due to fertility bottlenecks. These are<br />
expected to lead to genetic changes due to the twin factors of inbreeding and selection, with<br />
associated detrimental effects on the performance of released SIT flies. The purpose of the<br />
present experiment is to use microsatellite markers to investigate the relative importance of<br />
these two factors in the process of introducing wild strains into the laboratory.<br />
Methods: Eight populations were set up using flies collected from different areas of the Sydney<br />
region. These populations were sampled over ten generations. All samples were typed for nine<br />
of the most variable of the established Q-fly microsatellites. For six of the eight lines, 48 flies<br />
were sampled at generations 0, 1, 2, 3, 4, 6 and 10. For the remaining two lines, 48 flies were<br />
sampled at generations 0, 1 and 10. Data on the distribution of individual age-specific female<br />
fecundity were also collected.<br />
Results: The rate of inbreeding was shown to be highly repeatable. The observed inbreeding<br />
process was modelled by stochastic simulation of the random genetic drift occurring in the<br />
replicates line, taking into account the finite population size and distribution of family sizes.<br />
The model successfully predicted the rate of decline of heterozygosities in the replicate lines,<br />
showing that the inbreeding in new Q-fly lines is unavoidable, even in the absence of “hard”<br />
selection for increased early-age egg production. There was, however, also some evidence for<br />
selection as shown by repeatability of changes in two lines. Comparisons between factory flies<br />
and hybrids showed that it is possible to retain the high fertility of factory flies while<br />
improving performance in dispersal activity and stress resistance.<br />
Conclusions: Both selection and inbreeding are expected to have undesirable consequences in<br />
terms of the performance of SIT releases. This experiment indicates that inbreeding per se is a<br />
likely factor in the early generations of laboratory culture. It follows therefore, that measures to<br />
reduce the effects of inbreeding during the setting up of factory strains are of value. A protocol<br />
using four-way hybrids is outlined.<br />
Keywords: microsatellites, Q-fly, inbreeding<br />
71
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
A GENETICS COMPARATIVE WORK OF TWO DIFFERENT TEPHRITIDAE SPECIES,<br />
CERATITIS CAPITATA (WIEDEMANN) AND BACTROCERA OLEAE (GMELIN): RAPD<br />
DATA.<br />
Ochando, María Dolores*; Fernández, Paz; Segura, M. Dolores & Callejas, Carmen<br />
Departamento de Genética, Facultad de CC. Biológicas, Universidad Complutense, C/ José Antonio Novais, nº2,<br />
28040- Madrid, Spain; Email: dochando@bio.ucm.es.<br />
Background: The olive fruit fly, Bactrocera oleae, is a major pest of olive crops and its<br />
expansion is exclusively restricted to the cultivation zone of the olive tree. The medfly,<br />
Ceratitis capitata, however, is a highly polyphagous species that infest more than two hundred<br />
different vegetal species. Both constitute important pests in the Mediterranean area being<br />
responsible of important crops damages and economic losses. We report here a comparative<br />
work on the genetics structure of their populations based on RAPD molecular markers<br />
information.<br />
Methods: Flies from different geographical areas, mostly Mediterranean, were collected by<br />
harvesting infested fruit and allowing the larvae to pupate in the laboratory. Individuals from<br />
20 different locations for C. capitata and 21 for B. oleae, were analyzed on the basis of<br />
Random Amplified Polymorphic DNA through the Polymerase Chain Reaction (RAPD-PCR).<br />
Genomic DNA was extracted from individual flies according to Reyes, Ochando et al. (1997),<br />
and amplified with six or seven different primers (Sets A and C, Operon Technologies). DNA<br />
amplifications were performed under the conditions reported by Williams et al. (1990), with<br />
slight modifications. Each amplification reaction was performed at least twice: the results were<br />
consistently reproducible. The amplification products were separated according to their<br />
molecular size by electrophoresis in 2% agarose gels with TAE buffer.<br />
Results: The observed polymorphism was high in both species, even higher in B. oleae (0.49<br />
for Ceratitis and 0.66 for Bactrocera species). With respect to the distribution of the genetic<br />
variability, both species showed the same pattern. In both species the within population<br />
variation (between 76 and 90 %, AMOVA analysis) was significantly higher than the between<br />
populations variation. Genetics distances were low and similar for all North Mediterranean<br />
samples in both species without clear phylogenetics relationships. However the rest of the<br />
populations (Tunisia, Israel and USA for B. oleae, and Kenya, SouthAfrica, Tunisia and Israel<br />
for C. capitata) were clearly different.<br />
Conclusions: However the different ecological life of the two analyzed species, C. capitata and<br />
B. oleae, their genetic variabilities are roughly of the same level from the quantitative point of<br />
view and show similar characteristics from the qualitative point of view. Thus, the ecological<br />
and feed parameters can not explain those coincidences. Other aspects such as effective<br />
population numbers, gene flow and elapsed time from the colonization processes, must be<br />
taking into account for population structure in these pests.<br />
Keywords: C. capitata, B. oleae, RAPD-PCR, polymorphism, genetic distances.
Session 4<br />
Risk Assessment, Quarantine &<br />
Post-harvest Treatments
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
QUARANTINE SECURITY FOR COMMODITIES: DETERMINING THE NON-HOST<br />
STATUS OF FRUITS REGULATED FOR TEPHRITID FRUIT FLIES<br />
Liquido, Nicanor J.* 1 ; Hennessey, Michael K. 2 ; Griffin, Robert L. 2 ; Velapatiño, Jorge<br />
F. 3 ; Layme, Javier M. 3 & Gonzales, Luis B. 4<br />
1 United States Department of Agriculture (USDA)-Animal and Plant Health Inspection Service (APHIS)-Plant<br />
Protection and Quarantine (PPQ), Center for Plant Health Science and Technology (CPHST), 300 Ala Moana<br />
Blvd., POB 50002, Honolulu, Hawaii 96850 USA. Email: Nicanor.J.Liquido@aphis.usda.gov; 2 USDA-APHIS-<br />
PPQ, CPHST, Raleigh, North Carolina USA; 3 Ministro de Agricultura, Servicio Nacional de Sanidad Agraria<br />
(SENASA), Lima, Perú; 4 Asociacion Peruana de Productores de Palta Hass (ProHass), Lima, Perú.<br />
Background: Assessing the risk of fruit fly pests associated with the movement of various<br />
economically important commodities is paramount in facilitating global trade. Determining the<br />
fruit fly host status of a particular commodity is imperative in developing risk mitigation<br />
measures and establishing the intensity of quarantine treatments. Host suitability is a critical<br />
quarantine security parameter of several international and regional standards. In particular, we<br />
aim to review RSPM 30: “Guidelines for the Determination and Designation of Host Status of<br />
a Commodity for Fruit Flies (Diptera: Tephritidae).” Here we report on the parameters defined<br />
in RSPM 30 that we used in determining the host status of ‘Hass’ avocados in Peru to<br />
infestation by South American fruit fly, Anastrepha fraterculus (Diptera: Tephritidae).<br />
Methods: Experiments were conducted, following the RSPM 30 guidelines, to determine the<br />
host suitability of ‘Hass’ avocados to A. fraterculus under natural field conditions in five<br />
orchards located in Lima, Ancash and Piura. Both wild and laboratory-reared adults were used.<br />
The adult population density in test orchards was estimated by trapping and collecting natural<br />
host fruits. During the entire harvest season, commercial grade mature green ‘Hass” fruits were<br />
collected biweekly from trees in each test orchard, and the infestation level of A. fraterculus<br />
determined by peeling (removing skin and cutting pulp) and holding fruits in rearing<br />
containers. A. fraterculus infestation in fruits on the ground and culled fruits from packing<br />
houses was also determined. No-choice and choice oviposition tests using intact and punctured<br />
fruits were conducted in cages measuring 4-m high and 4-m diameter which covered an entire<br />
tree. No-choice forced oviposition tests were also conducted using sleeves measuring 1.5-m<br />
length and 1-m diameter that enclosed an entire branch. For oviposition tests, mangoes were<br />
used as control fruits.<br />
Results: Successful insertion of eggs by A. fraterculus into the skin (exocarp) and pulp<br />
(mesocarp) of mature green ‘Hass’ avocados was never observed in both choice and no-choice<br />
forced oviposition tests. All fruit samples collected from the field and packinghouses had no A.<br />
fraterculus infestation. The population density of A. fraterculus inside and outside the orchards<br />
was low.<br />
Conclusions: Based on the studies conducted following the guidelines of RSPM 30, mature<br />
green, commercial grade ‘Hass’ avocados appear to be conditional non-hosts of A. fraterculus,<br />
suggesting that the current mitigation options for A. fraterculus need to be evaluated based on<br />
the present results.<br />
Keywords: quarantine security, host suitability, systems approach; fruit flies; ‘Hass’ avocado<br />
75
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE POTENTIAL GLOBAL DISTRIBUTION OF NATAL FRUIT FLY, CERATITIS ROSA<br />
(KARSCH) (DIPTERA: TEPHRITIDAE).<br />
De Villiers, Marelize* 1 ; Hattingh, Vaughan 1 & Kriticos, Darren J. 2<br />
1 Citrus Research International (CRI), Department of Conservation Ecology & Entomology, Stellenbosch<br />
University, P.O. Box 2201, Matieland 7602- Stellenbosch (South Africa); Email: dev@sun.ac.za; 2 CSIRO<br />
Entomology, Canberra, Australia.<br />
Background: Ceratitis rosa is an important pest of various commercially grown fruit crops in<br />
South Africa. Its present distribution is restricted to Africa, Mauritius and Reunion.<br />
Phytosanitary restrictions are imposed on international fruit trade to mitigate against the risk of<br />
introducing this pest into importing countries. Knowledge of its potential to invade other parts<br />
of the world makes it possible to evaluate the relevance of such measures. The potential future<br />
global distribution of C. rosa has previously been modelled using correlative species<br />
distribution modelling techniques. However, these models were built on presence data and did<br />
not incorporate information on the species’ relative abundance and seasonal phenology. The<br />
wide range of climatic conditions in South Africa, and related differences in the species’<br />
abundance, makes the region well suited to obtain information on relative abundance and<br />
seasonal phenology of the species in relation to climate. The objective of the study was to<br />
model the potential global distribution of C. rosa based on its distribution, relative abundance<br />
and seasonal phenology in South Africa.<br />
Methods: Yellow bucket traps, bated with Biolure® Fruit Fly, were used to trap C. rosa in<br />
different climatic regions of South Africa. Traps were placed in host plants and serviced<br />
monthly for a two-year period. A CLIMEX niche model of the potential global distribution of<br />
C. rosa was fitted based on the collected trapping data and other distribution records from<br />
South Africa. Distribution records for elsewhere in Africa were reserved for independent<br />
model validation. CLIMEX is a generic dynamic simulation model that combines inferential<br />
and deductive modelling approaches. It can be used to infer what climatic conditions a species<br />
can tolerate based on where it lives or its phenology.<br />
Results: The CLIMEX model output conformed well to the data observed from trapping in<br />
South Africa, not only in terms of the species’ distribution, but also in terms of the relative<br />
abundance and seasonal phenology. The projected distribution also accords well with the<br />
pattern of presence records of the species elsewhere in Africa. The model predicts a large part<br />
of South America, Central America, Mexico and southern USA to be climatically suitable<br />
under recent climate conditions. In Europe, climatically suitable habitat is restricted to the<br />
coastal regions of the Mediterranean. In Asia, suitable climate is restricted mostly to the<br />
southern and south eastern countries, while in Australia it is mostly the wetter South and East.<br />
Conclusions: The independent cross-validation provided by relative abundance and seasonal<br />
phenology data, together with relevant species specific biological information enhanced the<br />
modelling of the species’ potential global distribution, thereby producing a more reliable<br />
model.<br />
Keywords: Biosecurity, CLIMEX, distribution, relative abundance, seasonal phenology
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ASSESSMENT OF THE RELIABILITY OF EXISTING PHYTOSANITARY MEASURES<br />
AGAINST THE INTRODUCTION OF NON-EUROPEAN TEPHRITIDAE INTO THE<br />
EUROPEAN COMMUNITY.<br />
Cobos-Suárez, José María 1 ; Guitián-Castrillón, José María* 2 & Catalán-Ruescas, Diana 2<br />
1 Ministerio de Medio Ambiente y Medio Rural y Marino (MARM), Subdirección General de Sanidad de la<br />
Producción Primaria, C/Alfonso XII 62; 28014 Madrid (Spain). Email: jcobossu@marm.es; 2 Tecnologías y<br />
Servicios Agrarios, S.A. (Tragsatec) C/Hnos. García Noblejas 37c, 2 nd floor; 28037 Madrid (Spain).<br />
Background: The Tephritidae family comprises 500 genera, including the Anastrepha,<br />
Bactrocera, Ceratitis and Dacus, and Rhagoletis genus, worldwide notorious for their<br />
destructive impact on agriculture. Except for Rhagoletis spp., which are found in temperate<br />
areas of Europe, few species of the other genera occur within the European Community (EC).<br />
As it includes transboundary pests of global concern, a key issue for pest risk analysts is to<br />
assess the reliability of the existing phytosanitary measures. We summarize here the current EC<br />
set of measures against non-European Tephritidae, identify their efficacy and compare them<br />
with those applied in third countries.<br />
Methods: The EC Plant Health Directive has been conscientiously revised in order to identify<br />
all references to Tephritidae and plant products with which the fruit flies have a possibility of<br />
being associated in a suitable life stage. We have checked the ability of fruit flies to survive the<br />
standard shipping methods by consulting the European interception database (Europhyt).<br />
Finally we have selected from the relevant pathways of introduction those which appear to<br />
pose a higher risk for the EC.<br />
Results: Two major pathways have been identified: (i) fresh fruits of host plant species and (ii)<br />
plants for planting (except seeds) accompanied by contaminated, attached growing media from<br />
countries where non-European Tephritidae occur. Frequently, only plants and plant products<br />
which are accompanied by a Phytosanitary Certificate are subject to a plant health inspection in<br />
the ports of entry. This common practice hinders the efficacy of the protective measures laid on<br />
the Directive and lead to visual inspection as the only requirement against fruit flies for most<br />
fruits (citrus are an exception). Regarding soil and growing medium attached, the recurring<br />
detection of harmful nematodes in these consignments shows the lack of implementation in<br />
practice of the requirements imposed to the EC trading partners.<br />
Conclusions: Visual inspection, as a single measure, is not enough to guarantee that fruits and<br />
plants with attached growing media are free from tephritids. This statement is supported by the<br />
regulation in force in third countries (e.g. USA, China, Australia or Japan) that do not rely<br />
solely on visual inspection of consignments. The EC, as major importer of plant products, is<br />
exposed to the entry of non-European Tephritidae and should impose stricter quarantine<br />
measures to prevent their spread within its territory.<br />
Keywords: Pest Risk Analysis, phytosanitary measures, reliability, pathway, introduction.<br />
77
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DESIGNING PRACTICAL LABORATORY PROCEDURES FOR DETERMINING HOST<br />
STATUS OF COMMODITIES TO FRUIT FLIES USING VARIOUS ANASTREPHA SPECIES<br />
AS EXAMPLES.<br />
Mangan, Robert L.* & Tarshis Moreno, Aleena<br />
USDA-ARS Crop Quality and Fruit Insects Research Unit, Kika de la Garza Subtropical Agricultural Research,<br />
2413 E. Hwy 83, Weslaco Texas, (USA); Email: robert.mangan@ARS.USDA.GOV.<br />
Background: A critical component of assessing pest introductions and requirement for<br />
quarantines in fresh fruits/vegetables is the host status that determines risk of pest transport and<br />
introduction. Here we present a summary of laboratory procedures that address host evaluation<br />
and illustrate the procedure with summaries of laboratory experiments with the Mexican fruit<br />
fly, Anastrepha ludens Loew, the West Indian fruit fly A. obliqua (Macquart), and the Sapote<br />
fly A. serpentina (Weidemann) in tests with various Citrus species and cultivars. Based on<br />
earlier tests, we classified host status as either natural hosts or conditional hosts, and fruit fly<br />
species were classified as natural pests or conditional pests.<br />
Methods: Experiments tested survival and development of eggs and larvae of the fruit flies to<br />
compare tissue specific survival of the developmental stages of these species in natural and<br />
conditional citrus hosts and determine the tissue specific survival rates related to the maturity<br />
of the fruit. Experiments were performed by exposing fruit in laboratory cages (30 cm cubic/ 8<br />
fruit per cage, 8 mature females/fruit) using fruit wrapped to permit oviposition in a marked<br />
area for 24 h. Tests comparing fruit fly species were run in separate cages concurrently. Fruit<br />
were unwrapped, held in containers for maturation, then samples were dissected after 3 d, and<br />
weekly for 4 weeks to determine tissue and time specific survival of 4 (egg, and 1 st -3 rd instar)<br />
stages in flavedo, albedo, and pulp.<br />
Results: In all tests most mortality occurred during the first week after oviposition for eggs and<br />
first stage larvae in the albedo. The Mexican and sapote fruit flies oviposited into the albedo<br />
but West Indian fruit fly eggs were found mostly in the flavedo among the oil glands. Results<br />
showed large differences among the species for factors such as fruit maturity and senescence<br />
affecting mortality in the albedo but mortality was very low in larvae reaching the pulp for any<br />
of the species.<br />
Conclusions: The methods described here allowed identification of factors that affect host<br />
status for the 3 Anastrepha species of high economic importance in North America. The<br />
method could be tested under laboratory conditions during a single season. The Mexican fruit<br />
fly which is an economic pest on most citrus (but not Eureka lemons) was most sensitive to<br />
fruit maturity. Fly trapping triggers for quarantines in areas using a systems approach to meet<br />
requirements could be adjusted based on these factors for conditional hosts.<br />
Keywords: host status, quarantine
Session 5<br />
SIT Principles & Application
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IMPROVING MASS REARING TECHNOLOGY FOR THE MEDFLY TEMPERATURE<br />
SENSITIVE LETHAL STRAIN (TSL): GEL LARVAL DIET DEVELOPMENT AND<br />
VALIDATION.<br />
Cáceres, Carlos.<br />
USDA-APHIS-PPQ-CPHST/MOSCAMED PROGRAM GUATEMALA. San Miguel Petapa Fruit Flies Mass<br />
Rearing and Research Center Facility. Calle Real 3-60, Zona 9, San Miguel Petapa, Guatemala. Email:<br />
carlos.e.caceres@aphis.usda.gov, caceres-ce@hotmail.com<br />
Background: Mass rearing technology is a crucial element to establish a functional Sterile<br />
Insect Technique (SIT) program for any species. Insect rearing represents approximately 50%<br />
of the SIT cost. A Gel larval diet and its rearing system for Medfly (Ceratitis capitata)<br />
production were developed at San Miguel Petapa Facility in Guatemala. Benefits of a Gel diet<br />
technology include a reduction in waste diet; elimination of the bulking agent and reductions in<br />
diet ingredient storage needs and labor.<br />
Materials and Methods: Medfly eggs for both male only and colony productions lines were<br />
produced at the El Pino Facility, Guatemala. Eggs batch samples (250 ml for colony and 700<br />
ml for male only) were divided into equal parts. Half of the samples were sent to the Petapa<br />
Facility where the eggs were transferred onto the Gel diet. The diet is composed of Torula<br />
yeast, sugar, antifungal agents (sodium benzoate), hydrochloric acid, tap water and organic gel<br />
agent. Five replicas were conducted once per week during 5 consecutive weeks. Each<br />
experiment included approximately 60 trays for colony and 60 trays for male only production.<br />
The trays were loaded with two kilos of diet then male only trays were infested with 5.2 ml of<br />
eggs while the trays dedicated to the colony were infested with 1.8 ml of eggs. At El Pino the<br />
control eggs were seeded following normal rearing procedures. Trays were loaded with 5 kg of<br />
corn cob diet; male only trays were infested with 6.4 ml of eggs while colony trays were<br />
infested with 3.2 ml of egg. In both Facilities maturing larva were collected and measured<br />
every day to determine larva production quality. During the pupal development stage samples<br />
were separated and the quality assessed using standard QC tests.<br />
Results: Larval rearing of Medfly on this gel diet resulted in 30% increase in pupal production<br />
while increases in the colony production line resulted in 47% more female pupal production as<br />
compared to the control diet. Pupal weight, adult emergence, adult fliers, egg hatch and egg<br />
production are basically equal to insects reared on a conventional corncob diet. This first mass<br />
rearing assessment has shown a significant saving (23%) on the total cost of diet to produce the<br />
same quantity of insects. While it is now expected that the Gel diet can save some diet related<br />
costs during the rearing process other operational components such as energy consumption,<br />
labor, space, etc. need to be considered in the overall calculation of cost savings.<br />
Conclusion: The utilization of Gel diet technology may allow for considerable cost savings.<br />
However more research will be necessary in the future to design the best equipment and<br />
facility configuration for this new technology. The diet formulation may need to be revised so<br />
that more savings could be achieved without compromising the quality of the insects.<br />
Keywords: Medfly, SIT, Medfly larval diet, Mass rearing<br />
81
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
METHODOLOGY AND SAMPLE SIZE TO ESTIMATE RECOVERED FLIES AND<br />
ABSOLUTE FLYER FLIES POST-CHILLING PROCEDURE OF STERILE MEDFLY,<br />
CERATITIS CAPITATA (WIEDEMANN).<br />
Villaseñor, Antonio 1 ; Gonzalez, Ricardo 1 ; Rasgado, Milton 1 ; Romero, Moises 1 ;<br />
Hernández, Emilio* 2 & Montoya, Pablo 2<br />
1 Programa Moscamed-DGSV-SENASICA-México, Centro de Empaque de Adulto en Frío. Carretera Tapachula-<br />
Puerto Madero Km 3.6, Chiapas, México; 2 Desarrollo de Métodos, Moscafrut-Moscamed SAGARPA-IICA. Calle<br />
Central Poniente No. 14, Centro. 30700. Tapachula, Chiapas, México. E-mail: emilioho@prodigy.net.mx.<br />
Background: The efficacy of the Sterile Insect Technique is based on the quantity and quality of the<br />
adults released, which could be determined by the percent of post- chilling recovered flies<br />
(quantity) and absolute flyer flies (quality) after shipment, packaging, emergence, feeding,<br />
chilling and collection processes. In this work we described the adapted methodology for the<br />
daily estimation of the two parameters mentioned above for sterile male flies of Ceratitis<br />
capitata in Tapachula, México.<br />
Methods: The evaluation was carried on using emergence tower type “Guatemala” conformed<br />
by 25 double-screen tap trays. In 20 trays we collocate ~25, 000 pupae that were previously<br />
weighted. The trays remained in the emergence room at 24 °C and 60% RH, with food in form<br />
of fine flour (90% sugar, 10% vegetable proteins) and water. On the fifth day, the trays were<br />
moved into a cold room to slow down adults at 0 – 3 °C for 30 minutes.<br />
To determine the recovered flies and absolute flyers, standard procedures using total content of<br />
each screen-tray were compared with the new proposed method, which consisted in a sample<br />
of 30 ml of chilled flies from each tray. We used the proportion 9:1 between the weight of<br />
adult fly and its puparia as a constant. Each sample was weighted and placed in a plastic<br />
container of 33 x 20.5 x 10.6 cm talc impregnated, which were collocated in open space at<br />
environmental temperature to stimulate the flies flying and to get the residue of non-flying<br />
flies. The percent of recovered flies was estimated from the difference of the initial average<br />
weight of pupa and the final weight or residue inside the containers. To calculate the sample<br />
size we used the formula nE = (S / Ex) 2 (FAO/IAEA/USDA 2003).<br />
Results: The average percent and SD of recovered flies was 80% ± 0.62, while absolute flyer<br />
flies was 91 % ± 1.21. These results were compared with those obtained from standard<br />
procedures, 88% and 91%, respectively. The average weight in the sample of absolute flyers<br />
was 7.24 ± 0.36; using the formula with an accuracy level of 0.1 (90% reliability), the<br />
estimated number of samples was (0.36/(0.0135*7.24)) 2 = 12.96, which means that number of<br />
samples can be reduced from 20 to 13.<br />
Conclusions: The developed methodology provides reliable information about quality of<br />
managed flies by the chilled adult release system, while avoiding the use of large numbers of<br />
flies that could be destined for release purposes.<br />
Keywords: Medfly, Ceratitis capitata, sample size, recovered flies, absolute flyers.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IMPROVING THE STERILE INSECT TECHNIQUE THROUGH SELECTION OF BEST<br />
PERFORMING MALES.<br />
Liedo, Pablo 1 *; Hendrichs, Jorge 2 ; Toledo, Jorge 1 ; Oropeza, Azucena 1 ; & Jessup,<br />
Andrew J. 2<br />
1 Departamento de Entomología. El Colegio de la Frontera Sur. ECOSUR, Carretera Antiguo Aeropuerto Km 2.5,<br />
Tapachula, Chiapas, México 30700; Email: pliedo@ecosur.mx; 2 Insect Pest Control Subprogramme, Joint<br />
FAO/IAEA Division, Vienna, Austria.<br />
Background: The Sterile Insect Technique (SIT) requires mass-production of sterile insects and<br />
mass-rearing is a selection process. There is evidence of favorable traits that have been<br />
inadvertently selected through colonization and mass-rearing, such as fast development, high<br />
fecundity, ovipositing in egging devices, and large size. However, there is also evidence of<br />
non-intentional negative selection. For example, selection for fast mating, the loss of predator<br />
evasion ability and requirements for high light intensity and temperatures. Our goal in this<br />
study was to evaluate if selection based on survival ability, predator avoidance and mating<br />
competitiveness can improve the performance of mass-reared flies for SIT application, and if<br />
this approach could be incorporated into new colony management strategies of ‘mother stocks’.<br />
Methods: Strains of two mass-reared species were evaluated, the Mexican fruit fly, Anastrepha<br />
ludens, and the Mediterranean fruit fly Ceratitis capitata. Selection was done on host plants<br />
under field cage conditions. Wild flies were used as controls. Males selected by their survival<br />
ability, predator avoidance and wild female mating choice were crossed with mass-reared<br />
females and their offspring was reared following standard laboratory protocols. Offspirng<br />
performance in the same traits was evaluated in the next generation.<br />
Results: After one selection cycle we did not observe significant improvements in the general<br />
performance of males of the two species. Differences in survival and predator avoidance were<br />
not significant. However, the number of matings achieved by selected males were significantly<br />
greater than those achieved by non-selected (control) males.<br />
Conclusions: The slightly better mating performance of selected males was consistently found<br />
in both species. Possible reasons for the lack of more significant effects will be discussed and<br />
new research lines will be proposed. The feasibility of incorporating such selection processes<br />
in the routine colony management of “mother stocks’ is analyzed.<br />
Keywords: Anastrepha ludens, Ceratitis capitata, mass-rearing, sterile males<br />
83
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
NINE YEARS OF MASS-REARED OF WEST INDIAN FRUIT FLIES ANATREPHA<br />
OBLIQUA (MACQUART) DIPTERA: TEPHRITIDAE IN THE MOSCAFRUT COMPLEX.<br />
Orozco-Dávila, Dina; Artiaga, Trinidad*; Hernández, M. Refugio & Domínguez, Julio<br />
Planta Moscafrut SAGARPA-IICA. Central Poniente No. 14. C.P. 30700. Tapachula, Chiapas. México. E-mail:<br />
tartiagal@hotmail.com<br />
Background: Anastrepha obliqua is the second most economically important species for<br />
mango Mangífera indica L. growing in México (Aluja et al. 1987). One of the methods for<br />
controlling this pest is the Sterile Insect Technique (SIT). The Moscafrut mass rearing facility<br />
has produced this species. Since 2001, the rearing process has been optimized. In 2007 it was<br />
possible to rear the mother colony under more relaxed conditions and this improved the quality<br />
of the flies and male sexual competitiveness. In 2009 the highest competitiveness index was<br />
achieved.<br />
Methods: The rearing conditions have involved loading cages with 60,000 pupae for the relax<br />
colony and 88,000 for release. Environmental conditions have been: 26 ±1°C and 70±5 RH.<br />
Larval density in the diet has been 3.7 and 5.58 larvae/ per g of diet, for relax and release<br />
rearing, respectively. The diet formulation has been: 16.33% corncob powder, yeast 6.33%,<br />
8.66% corn flour, sugar 9.0%, 0.1% guar gum, Nipagin 0.18%, 0.33% sodium benzoate and<br />
citric acid 0.43%. The larvae are maintained for 3 days at 26-27 ° C, and then transferred to a<br />
room at 25 to 26 °C for 3 days and finally kept at 24-25 ° C until completing eight days in<br />
total. The mature larvae are separated by dissolving the diet in water. Pupation takes place at<br />
20 ± 1 °C for 24 hours, then the pupae are maintained at 26°C and 80% RH for 13 days.<br />
Finally, the pupae for the colony are selected and the rest is irradiated at 80 Gray and sent to<br />
release areas in Mexico.<br />
Results: From 2001 to 2009 egg production increased from 0.416 to 1.14 eggs/cage/day.<br />
Fertility increased from 82.43% to 89.32%. Egg to larva survival has increased from 57.73% to<br />
73.07%. Mean larva weight in the relax rearing has been 19.12 mg, and 18.71 for the release<br />
rearing. Pupae production increased from 262.649 to 2,286.885 million per year. Mean pupa<br />
weight has been 14.11 and 13.75 mg for relax and release rearing, respectively. The percentage<br />
of adult emergence and fliers gradually increased, reaching values of 86.66% and 79.82%<br />
respectively. With the implementation of the relax colony in 2007, the sexual behavior of the<br />
produced males has been successful. The RSI increased to 0.42, which has been the highest<br />
value. Competitiveness with wild flies has been acceptable with 0.63 competitiveness<br />
coefficient, above the acceptable values for the coefficient competitiveness parameter for<br />
sterile males (0.2-0.4 FAO/IAEA 2003).<br />
Conclusions: The Anastrepha obliqua mass-reared system has improved during the past 9<br />
years, accomplishing the goals of the National Campaign, both in quantity and quality.<br />
Keywords: mass-reared, Relative Sterile Index (RSI) and competitiveness.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EVALUATION OF GENETIC SEXING STRAIN “TAPACHULA-7” (T (Y-BP + )-7) OF<br />
ANASTREPHA LUDENS (LOEW) FOR STERILE INSECT TECHNIQUE.<br />
De León-Crisóstomo, Ángel H.; García-Martínez, Víctor; Ibañez-Palacios, Jorge; Flores-<br />
García, Hector; Meza, J. Salvador & Zepeda-Cisneros, Cristina S. *<br />
National Fruit Fly Campaign. Moscafrut Program. Acuerdo SAGARPA-IICA. Central Poniente No. 14 Col.<br />
Centro Tapachula Chiapas, México. Email: cczepeda@hotmail.com<br />
Background: Genetic sexing strains (GSS) can significantly enhance Sterile Insect Technique<br />
applicability and efficiency, for Fruit Flies are becoming more relevant every day due to their<br />
operational advantages and substantial saving that represents for the action programs. A GSS<br />
based on the Y-autosome translocation system has been developed for A. ludens; typically<br />
pupaes are brown/males: black/females. This strain has been named “Tapachula-7” and after<br />
making small-scale assessments has been identified a “good potential” for semi-massive<br />
scaling. We report here the evaluation on viability and stability in a production that is being<br />
increased.<br />
Methods: A stock colony of the GSS “Tapachula-7” was maintained at densities of 1500 pairs<br />
of flies per cage each generation; viability parameters were measured and screened the total<br />
production recording the number of aberrants. Furthermore other cages has been maintained at<br />
densities of 3000-4000 pairs per cage, the total flies produced were revised each generation<br />
recording since F8 the crippled/half emerged flies. Production parameters were estimated in<br />
order to calculate the rearing efficiency.<br />
The genetic study of aberrants were done by means of individual crosses, each aberrant fly was<br />
crossed with a “wild type” insect first, and then with a "black pupae” mutant. Their offspring<br />
was recorded for two generations.<br />
Results: The stock colony “Tapachula-7” has been maintained during 19 generations and no<br />
aberrants has been found. The fertility has been maintained in 70%, which is a good value for a<br />
strain carrying a Y-autosome translocation. The viability was measured as percentage of<br />
transformations at different developmental stages: larvae-brown pupae=33.56; larvae-black<br />
pupae= 35.56, brown pupae-adult=92.34, black pupae-adult=76.59. The production was scaled<br />
and actually we maintain 5 cages with 3000 pairs each one. From these cages we obtained in<br />
average 10ml egg/day/cage and a production of pupaes of 9,000/day/cage. The percentage of<br />
crippled and half merged flies was less than 2%. Only “black pupae” male aberrants have been<br />
found, when we crossed individually these insects with wild type females, the F1 progeny was<br />
only wild type insects, but in F2 appeared “black pupae” and wild type flies. In these aberrants<br />
insects the crosses with “black pupae” females did not produce any offspring.<br />
Conclusions: Production and stability parameters of Genetic Sexing Strain “Tapachula-7”<br />
allow for reliable predictions on that the strain will perform properly under mass rearing<br />
conditions.<br />
Keywords: mexicanfruitfly, genetic sexing strain, Anastrepha ludens, translocation, new strain evaluation, SIT<br />
85
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
JUVENILE HORMONE TREATMENT AND PROTEIN SUPPLY IN ADULT DIET: THE<br />
CASE OF ANASTREPHA FRATERCULUS.<br />
Segura, Diego 1,2* ; Vera, M. Teresa 2,3 ; Liendo, M. Clara 1 ; Devescovi, Francisco 1 ; Utgés, M.<br />
Eugenia 4 ; Abraham, Solana 3 ; Peralta, Patricia 1 ; Carabajal Paladino, Leonela 1 ;<br />
mann, Guillermo 1 ; Milla, Fabián 1 ; Teal, Peter 5 & Cladera, Jorge L. 1<br />
1 Instituto Nacional de Tecnología Agropecuaria, Buenos Aires, Argentina. Los Reseros y Repetto, Castelar<br />
(1712), Buenos Aires, Argentina. dsegura@cnia.inta.gov.ar; 2 CONICET, Argentina. 3 Estación Experimental<br />
Agroindustrial Obispo Colombres, S. M. de Tucumán, Argentina. 4 Facultad de Ciencias Exactas y Naturales,<br />
Universidad de Buenos Aires, Argentina; 5 Center for Medical, Agricultural, and Veterinary Entomology, USDA-<br />
ARS, Gainesville, FL, USA.<br />
Background: Anastrepha fraterculus is a major fruit pest in South America. Our research<br />
focuses on obtaining baseline data for the implementation of the Sterile Insect Technique. In<br />
this species, males have a long pre-copulatory period. We studied the effect of treating A.<br />
fraterculus with a juvenile hormone analogue, methoprene, which accelerates maturation in<br />
other Anastrepha. Because sexual development is tightly associated to protein intake,<br />
methoprene effects were analyzed under several nutritional regimes.<br />
Methods & Results: In laboratory, we found that methoprene treatment allowed a faster sexual<br />
maturation of A. fraterculus fertile as well as sterile (gamma-irradiated) males. Further studies<br />
showed no differences in transferred sperm between methoprene-treated males and mature,<br />
untreated males. Protein-fed, methoprene-treated males reached sexual maturity earlier than<br />
treated males that fed on sugar, or untreated males fed on sugar plus protein. However, this was<br />
not the case for every protein source; hydrolyzed yeast showed the highest percentage of<br />
matured males. We found that a 12:1 (sugar: hydrolyzed yeast) diet is enough for methoprene<br />
to shorten the pre-copulatory period. Under an SIT program, methoprene must be delivered to<br />
millions of males at once. Dipping pupae in a methoprene solution allowed males to mature as<br />
fast as topically treated males. Including methoprene in adult diet also induced an accelerated<br />
development, but it relies on methoprene solubility in water. In field cages, we found that 6<br />
days-old laboratory males treated with methoprene (by dipping pupae in methoprene) and fed<br />
sugar and hydrolyzed yeast at a 12:1 ratio were able to courtship, to attract of females, to mate,<br />
to transfer sperm and to induce a refractory period for mating in females as well as sexually<br />
mature wild males fed on a 3:1 diet. Young laboratory females showed lower performance<br />
(only 5% mated) than wild females irrespectively on the fact that they were treated with<br />
methoprene and fed a diet containing protein.<br />
Conclusions: Sexual maturation of A. fraterculus sterile males can be accelerated by treating<br />
the flies with methoprene, but protein is needed for methoprene to act. Methoprene and protein<br />
allow laboratory males to compete for wild females in field cages. Massive delivery of<br />
methoprene is possible through feeding and dipping, and future studies should focus on<br />
massive delivery of protein and sugar. In the absence of a genetic sexing system, the fact that<br />
females do not respond as males to methoprene acts as a physiological sexing effect.<br />
Keywords: South American fruit fly, SIT, sexing system, nutrition.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECTS OF A JUVENILE HORMONE ANALOGUE, METHOPRENE, AND DIETARY<br />
PROTEIN ON BACTROCERA CUCURBITAE (COQUILLETT): IMPLICATIONS FOR SIT.<br />
Haq, Ihsan ul *1 ; Cáceres, Carlos 1 ; Hendrichs, Jorge 2 ; Teal, Peter 3 & Robinson, Alan S 1<br />
1 Insect Pest Control Laboratory, Joint FAO/IAEA Agriculture and Biotechnology Laboratories, A-2444 Seibersdorf,<br />
Austria. E-mail: I.Haq@iaea.org; 2 Insect Pest Control Section, Joint FAO/IAEA Division, IAEA, Wagramerstrasse<br />
5, A-1400 Vienna, Austria. 3 Center for Medical, Agricultural, and Veterinary Entomology, ARS,<br />
USDA, 1600-1700 SW 23rd Drive, Gainesville, Florida, USA.<br />
Background: Juvenile hormone analogue, methoprene treatment can reduce the time required<br />
for sexual maturation and access to diet including protein enhanced mating success in male<br />
melon fly, Bactrocera cuccurbitae (Diptera: Tephritidae) genetic sexing strain under laboratory<br />
conditions, supporting their use as a treatment for sterile males within the context of sterile<br />
insect technique (SIT).<br />
Methods: The effect of access to dietary protein (P) (hydrolyzed yeast) and/or treatment with a<br />
juvenile hormone analogue, methoprene (M), (in addition to sugar and water) on B. cucurbitae<br />
was assessed in the laboratory and in field cages. The males were exposed to either 1) protein<br />
and methoprene (M+P+), 2) only protein (M-P+), or 3) only methoprene (M+P-), and<br />
compared with untreated males (M-P-). The effect of methoprene and protein on mating<br />
behaviour of the four groups of males, when competing for virgin sexually mature females (M-<br />
P+), on starvation survival and on total body protein and lipids was studied.<br />
Results: Access to a diet including protein significantly increased male performance at leks and<br />
mating success as compared to only sugar fed males. Application of methoprene had a<br />
significant effect on accelerating sexual maturity only in males with access to a diet including<br />
protein, but not in males feeding only on sugar. Combined exposure to methoprene and protein<br />
also showed a significant effect on male performance at leks. More M+P+ males called and<br />
initiated and participated in lek activities than all other types of male, attaining a higher mating<br />
success. No adverse effect of methoprene and/ or protein on starvation survival was observed<br />
when compared with the untreated males. Treating males with methoprene and/ or protein<br />
didn’t inhibit female re-mating, however female mated with protein- or sugar-fed males during<br />
their first mating always preferred to re-mate with protein-fed males. Protein incorporation into<br />
the diet significantly increased the male body weight, total body carbon (TBC) and total body<br />
nitrogen (TBN) as compared to only sugar-fed males. Methoprene had no significant effect on<br />
males’ TBC and TBN.<br />
Conclusions: These studies provide strong evidence for the benefits of methoprene application<br />
and protein incorporation into the adult diet of sterile males. Treated males achieve higher<br />
sexual success, reach sexual development several days earlier, and are therefore much closer to<br />
sexual maturity when released as part of SIT action programmes after being held in the fly<br />
emergence and release facility.<br />
Keywords: Melon fly, methoprene, dietary protein, sexual success, sterile insect technique (SIT)<br />
87
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IMPROVE MATING SUCCESS OF YOUNG STERILE MALES OF BACTROCERA<br />
DORSALIS (HENDEL) AND BACTROCERA CORRECTA (BEZZI).<br />
Orankanok, Watchreeporn*; Chinvinijkul, Suksom; Pinkaew, Supaap & Brohmsubha,<br />
Suttipun<br />
Department of Agriculture Extension, Ministry of Agriculture and Cooperatives, 2143/1 Phahol Yotin Road,<br />
Chatuchak, Bangkok, 10900, Thailand. Email: watchreporn@doae.go.th; Watchreporn@yahoo.com<br />
Background: Two tephritid fruit flies (Diptera: Tephritidae), the Oriental fruit fly, Bactrocera<br />
dorsalis (Hendel), and the guava fruit fly, Bactrocera correcta (Bezzi), are two key insect pests<br />
of fruits production causing yield loss, quality degradation and restrict international trade in<br />
Thailand and some Asia-Pacific countries. The Sterile Insect Technique, SIT, has been<br />
implement to control fruit fly species in pilot areas of mango, Mangifera indica Linn.<br />
(Anacardiaceae). Up-scaling from a pilot areas project to a country-wide level needed to apply<br />
SIT in a cost-effective way.<br />
Metods: Two prerelease diet sugar, S, only and combination of sugar and protein hydrolysate,<br />
SP 3:1, were provided to sterile males during emerging, sexual separation and continuously<br />
until 2 and 3 day-old then followed with synthetic methyl eugenol, ME (1,2-dimethoxy-4-(2propenyl)<br />
benzene), exposed and not exposed at releasing age 2 and 3 days. Mating<br />
competitiveness experiments were conducted as 2x2 factorial in RCBD, (S without ME); (S<br />
with ME); (SP 3:1 without ME) and (SP 3:1 with ME) over five replications. Sterile males<br />
(2+n and 3+n, where n = 1-15 testing day) of B. dorsalis were set up; 50 sterile males and 50<br />
wild males, 23 day-old, were completed for 50 wild females, 21 day-old. But the observation<br />
day for B. correcta was 18 days. The same number of B. correcta sterile and wild males were<br />
designed for wild females competition as B. dorsalis but the wild females was 37 day-old.<br />
Results: Bactrocera dorsalis and B. correcta showed additive interaction of ME and both S and<br />
SP 3:1 as prerelease diet via significantly higher percent of mating. RSI of sterile males given<br />
S with ME and SP with ME presented higher the percentage of mating significantly than<br />
prerelease diet feeding without ME exposure.<br />
Conclusions: Based on our result, we would recommend for weekly sterile male released<br />
program. Sterile males should provide sugar/protein (3:1) into adult eclosion box for 2-3 days<br />
then they would be exposed to ME in the morning of release day.<br />
Keywords: Bactrocera dorsalis, Bactrocera correcta, mating success, prerelease diet, methy eugenol.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF TRANSPORTATION AND SHIPPING IN THE QUALITY OF THREE MASS<br />
REARED SPECIES IN MEXICO.<br />
Hernández, M. Refugio*; Bravo, J.; Flores, S.; Domínguez, Julio & Orozco-Dávila, Dina<br />
Programa Operativo Moscafrut. Central Poniente No. 14 altos-Esq. 2ª Avenida Sur. CP 30700. Tapachula,<br />
Chiapas, MÉXICO.<br />
Background: The transport and shipment of fruit flies to be used in the Sterile Insect Technique<br />
(SIT) have to consider two basic aspects: 1) administrative and operational requirements and 2)<br />
Handling of biological material which includes type and conditions of packaging,<br />
transportation, distance traveled (shipping time), management in emergency areas, system for<br />
emergency and finally the release of biological material. The conditions that remain may affect<br />
the quality of adults produced, but even with optimal management, shipping time affect the<br />
emergence and flight ability of adults and negative effects in the performance of insects,<br />
competitiveness of flies or search capacity of parasitoids.<br />
Methods: Moscafrut Facility located in the south-east of Mexico have a mass rearing of<br />
Anastrepha ludens, A. obliqua and D. longicaudata. The biological material produced (sterile<br />
pupa or parasitized pupa) is separated from the substrate of pupation and packed in sausage<br />
bags, which are placed in cardboard boxes are then transported by air to the International<br />
Airport of Mexico City where they are transshipped to packaging and releases centers in the<br />
Northern states. During the packing, the pupae are maintained under hypoxic conditions for the<br />
sterilization of the flies, preventing the emergence of the material during the transportation. To<br />
measure the effect of shipping on the quality of both flies and parasitoid we compare the<br />
information generated during three years at origin and destination.<br />
Results: The transportation and shipment of the pupa to the destination resulted in a significant<br />
loss in quality parameters for each species distributed in the different states of Mexico. To<br />
Anastrepha ludens, the emergency of 92.78% and fligth ability of 90.37% were reduced by<br />
6.78% and 11.21% respectively. For A. obliqua, sent to two states, the values were 91.90% and<br />
86.51% with 2.63 and 6.34% loss for emergency and flight ability, respectively. The<br />
emergence of the parasitoid, D. longicaudata, was decreased by 9.14% at arrival with an initial<br />
value of 60.55%. The quality was affected significantly by the time of hypoxia, the effect being<br />
most noticeable when observing differences in travel times within a single destination.<br />
Conclusions: To minimize the effects on the quality of mass-produced insects, logistics must<br />
be organized in order to reduce the time of hypoxia.<br />
Keywords: emergence, flight ability, hypoxia, Anastrepha, Diachasmimorpha<br />
89
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
OPTIMUM DOSE OF GINGER ROOT OIL TO TREAT STERILE CERATITIS CAPITATA<br />
(WIEDEMANN) (DIPTERA: TEPHRITIDAE) MALES.<br />
Paranhos, Beatriz J *1 ; McInnis, Donald 2* ; Morelli, Renata 3 ; Castro, Rosemary M 1 ;<br />
Garziera, Luiza 1 ; Paranhos, Lucas G 1 ; Costa, Karen 4 ; Gava, Carlos 1 ; Costa, Maria de<br />
Lourdes Z 4 & Walder, Julio M M 4<br />
1 Laboratory of Entomology, Embrapa Semi Arid, C.P. 23, 56302-970, Petrolina-PE, Brazil. E-mail:<br />
bjordao@cpatsa.embrapa.br; 2 USDA-ARS Pacific Basin Agricultural Research Center, Honolulu-HI, USA; 3 Dept.<br />
of Entomology, Phytopathology and Agricultural Zoology-University of São Paulo (ESALQ), Piracicaba-SP,<br />
Brazil; 4 Laboratory of Food Irradiation and Radio-Entomology, University of São Paulo (CENA), Piracicaba-SP,<br />
Brazil.<br />
Background: The sterile insect technique (SIT) is widely used as part of an integrated approach<br />
to reduce field populations of the Mediterranean fruit fly (Ceratitis capitata, Diptera:<br />
Tephritidae). Aromatherapy based on exposure to ginger root oil (GRO) volatiles is known as a<br />
method to significantly improve the sexual performance of sterile medfly males, and is being<br />
used in many mass-rearing facilities around the world. However, the optimum dose of GRO is<br />
not well defined.<br />
Methods: This work evaluated in laboratory cages four different doses of GRO and a control<br />
(0, 0.1, 0.25, 0.5, and 0.75 ml/m 3 ), and two different methods to hold flies prior to release<br />
(paper bags and plastic cages). The objective was to find the lowest dose that provides optimal<br />
improvement in mating performance of sterile males when competing with wild males for wild<br />
females and optimal reduction in fertility of wild females. Egg hatch, copula duration, the<br />
Relative Sterility Index (RSI), and a Competitiveness (C) value (based on RSI) were calculated<br />
for each treatment to assess male sexual performance and induction of sterility.<br />
Results: The method used to hold flies did not influence the aromatherapy effect. The mean<br />
time spent by wild females in copula with wild males was significantly longer than with sterile<br />
males for all treatments, except when sterile males were treated with 0.1 ml of GRO/m 3 .<br />
Among all doses studied, the dose 0.1 ml of GRO/m 3 reached the highest levels for both RSI<br />
and induced sterility, and was not statistically different from the 0.25 and 0.5 GRO/m 3 doses.<br />
Conclusion: The flies can be treated aromatically with ginger root oil in either paper bags or<br />
ventilated plastic cages. It is recommended to apply the lowest dose, 0.1 ml of GRO/m 3 ,<br />
because it showed the best cost/benefit ratio when used in the Mediterranean fruit fly SIT<br />
programme in the San Francisco River Valley, Brazil.<br />
Keywords: Aromatherapy, Sterile insect technique, Mediterranean fruit fly, Medfly, GRO.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FIELD CAGE STUDIES ON MATING ISOLATION BETWEEN SPECIES WITHIN THE<br />
BACTROCERA DORSALIS COMPLEX.<br />
Jessup, Andrew J.* 1 ; Schütze, Mark K. 2 ; Clarke, Anthony R. 2 ; Islam, Amirul 1 &<br />
Wornoayporn, Viwat 1<br />
1 Insect Pest Control Laboratory. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture.<br />
International Atomic Energy Agency, Vienna, Austria; Email: a.jessup@iaea.org; 2 Discipline of Biogeosciences<br />
& CRC National Plant Biosecurity, Queensland University of Technology, G.P.O. Box 2434, Brisbane 4000,<br />
Queensland, Australia.<br />
Background: There is increasing demand, world wide, to address the issue of fruit fly species<br />
complexes, i.e. fruit fly species with distinct similarities that have been grouped together.<br />
There is concern that some of these insect lines are not separate species but geographical<br />
variants. Their uncertain taxonomic status is having significant implications for international<br />
trade and the efficacious use of the Sterile Insect Technique (SIT).<br />
Methods: Interspecific crosses between the following species were assessed: Bactrocera<br />
dorsalis, B. philippinensis, B. invadens and B. carambolae. 1) No-choice laboratory mating<br />
tests. Fly species were reared under identical conditions, sexed within 24h of adult emergence<br />
and housed in small cages with food and water. Five male flies of one species and 5 females of<br />
another species were housed in each cage and the experiment was replicated three times. Data<br />
on the numbers of eggs oviposited, egg fertility, egg-pupa recovery and the sex-ratio of F1<br />
hybrids were recorded. 2) Field cage mating tests. Flies were segregated by sex and “painted”<br />
with a dot of water-based paint when sexually mature. Each species / gender was painted with<br />
a different colour. Twenty males of each of the two species being tested were released into<br />
each of four field cages and, 30min later, 20 females of each of the two species were released.<br />
When mating couples formed the time of coupling was recorded, the mating couple was<br />
removed and identified through their colour code.<br />
Results: Under no-choice laboratory mating tests all attempted hybridisations produced viable<br />
offspring. However, under simulated field conditions various degrees of mating isolation<br />
between species were observed.<br />
Conclusions: Implications of these observations are discussed in regard to species separation,<br />
international trade and SIT.<br />
Keywords: Bactrocera dorsalis, species complex, mating behaviour, interspecific hybridisation<br />
91
Session 6<br />
Area-Wide & Action Programs
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
HAWAII AREA-WIDE FRUIT FLY INTEGRATED PEST MANAGEMENT PROGRAM: A<br />
MODEL SYSTEM.<br />
Vargas, Roger* 1 ; Mau, Ronald 2 ; Jang, Eric 1 & Wong, Lyle 3<br />
1 USDA, ARS, Pacific Basin Agricultural Research Center, P. O. Box 4459, Hilo, HI (USA). Email:<br />
roger.vargas@ars.usda.gov; 2 University of Hawaii at Manoa, Cooperative Extension Service, 3050 Maile Way,<br />
Gilmore 310, Honolulu, HI 96822; 3 Hawaiian Department of Agriculture, Division of Plant Industry, 1428 South<br />
King St., Honolulu, HI 96814.<br />
Background: In Hawaii, fruit flies limit development of a diversified fruit and vegetable<br />
industry, require export fruits to undergo expensive quarantine treatments, and provide a<br />
reservoir for pest introduction into the mainland United States. Hawaii has four species that<br />
have become established, three of which (Mediterranean fruit fly, Ceratitis capitata<br />
(Wiedemann), melon fly, Bactrocera cucurbitae (Coquillett), and oriental fruit fly, Bactrocera<br />
dorsalis (Hendel)) are considered major pests of agriculture worldwide. When these pests are<br />
introduced into the U.S. mainland, they often require large-scale eradication programs, often at<br />
great public expense. In California, where the total value of the fruit and vegetable industry has<br />
been estimated to be more than $14 billion annually, the California Department of Food and<br />
Agriculture has estimated that an established infestation of Mediterranean fruit fly would cost<br />
from $855 million to $1.4 billion during the first year of establishment.<br />
Methods: A partnership Hawaii Area-Wide Pest Management (AWPM) project implemented<br />
by ARS resulted in the first successful program to control fruit flies that have been devastating<br />
Hawaiian agriculture for almost 100 years. The control system based on a combination of<br />
techniques (sanitation, protein baits, male annihilation, and SIT), developed primarily by ARS,<br />
have been adapted and coordinated into an IPM initiative specifically designed to work in<br />
Hawaii’s environment. A hallmark of the program has been a network of partnerships<br />
involving ARS, the Hawaii Department of Agriculture, the University of Hawaii Cooperative<br />
Extension Service and local communities, with the support of APHIS and other research,<br />
regulatory and government agencies.<br />
Results: New technologies transferred to farmers included new monitoring systems, new<br />
protein bait systems, and new male annihilations treatments. The 2,079 signed cooperating<br />
growers in the program, representing 6,798 acres, 491 farms, across five islands so far-Oahu,<br />
Hawaii, Molokai, and Maui--have already been able to cut conventional pesticide use by 75-<br />
90%, and reduced fruit fly infestation from 30-40% to less than 5%. Small farms are now<br />
growing crops they had previously been abandoned due to fruit fly damage.<br />
Conclusions: An economic assessment determined that the Hawaii AWPM program was easy<br />
to use, and initial economic benefits were estimated at $2.6 million per year and projected.<br />
Even using the most conservative economic analysis, without including the possible benefits<br />
category, the rate of return still came to 27%, according to a cost-benefit study. The success of<br />
the Hawaii AWPM program has had international impacts on fruit fly management, as many<br />
other countries facing similar problems. Researchers and officials from People’s Republic of<br />
China, the Commonwealth of the Northern Mariana Islands, French Polynesia, Guam, Senegal,<br />
and Taiwan, among others, have expressed interest in or adopted the program as a model for<br />
fruit fly suppression.<br />
Keywords: Mediterranean fruit fly, oriental fruit fly, melon fly, area-wide integrated pest management.<br />
95
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
STATUS OF BACTROCERA INVADENS MANAGEMENT IN AFRICA: RECENT<br />
ADVANCES, SUCCESSES AND CHALLENGES.<br />
Ekesi, Sunday 1,* ; Vayssières, Jean-François 2 ; Hanna, Rachid 3 ; De Meyer, Marc 4 ;<br />
Goergen, Georg 5 & Malacrida, Anna 6<br />
1 ICIPE, PO Box 30772-00100, Nairobi, Kenya; Email: sekesi@icipe.org; 2 IITA-CIRAD,Cotonou, Benin; 3 IITA,<br />
Yaoundé, Cameroon; 4 RMCA, Tervuren, Belgium; 5 IITA, Cotonou, Benin; 5 Università di Pavia, Italia.<br />
Background: Bactrocera invadens was first detected at the Kenyan coast in 2003 and later<br />
described as a fruit fly pest that was completely new to science. It is probably native to Sri<br />
Lanka and has rapidly expanded its geographical range, now reported from over 28 African<br />
countries. The pest attacks numerous cultivated and wild host plants but mango is the primary<br />
host with damage ranging from 40-80%. Quarantine restrictions due to the insect have<br />
significantly restricted access to lucrative export markets abroad. For example, exports of host<br />
crops of B. invadens are already banned to Seychelles, Mauritius and South Africa; and trade<br />
of several horticultural produce between Africa and the US has been severely hampered.<br />
Methods: Since its detection, several international and national partners have been involved in<br />
investigations related to its distribution, abundance, host range and interaction with native<br />
Ceratitis species. Morphometrics and molecular studies on the insect have been carried out to<br />
unravel its relationship with other Bactrocera species as well as its invasion history. Several<br />
attractants and fungal-based biological control agents have been evaluated for suppression.<br />
Assessments of the role of the weaver ant Oecophylla longinoda in significantly decreasing<br />
damage, as well as farmer perception of the ant, have been carried out. Exploration for natural<br />
enemies in the putative aboriginal home has been undertaken and parasitoids have been<br />
introduced into Africa from established laboratories from the USA. Releases of Fopius<br />
arisanus has commenced in several countries. Post harvest disinfestations trials on citrus and<br />
avocado has been initiated jointly with the private sector to recoup export markets.<br />
Results: A comprehensive assessment of the distribution of B. invadens in Africa and potential<br />
range of expansion based on ecological niche model will be presented. Field observation<br />
depicts rapid displacement of native Ceratitis species. Host range studies showed that the pest<br />
attacks over 40 plants species. Morphometric analysis shows clear relationships with<br />
Bactrocera kandiensis and ongoing DNA barcode analysis will be presented. Analysis of<br />
invasion history using microsatellite markers revealed 2 points of invasion in Africa – Kenya<br />
and Benin. Potent attractants (e.g. GF-120 and Mazoferm, local waste brewer’s yeast) and<br />
fungal-based biopesticide have shown considerable potential for field suppression. Field<br />
releases of Fopius arisanus have commenced in several countries with significant recoveries<br />
indicating potentials for establishment. Results from ongoing cold disinfestations trials on<br />
avocado and citrus will be discussed.<br />
Conclusions: An IPM package based on the use of protein bait, male annihilation technique,<br />
biopesticides, biological control (ants and parasitoids) and orchard sanitation is being<br />
advocated, and progressively implemented by growers for access to both domestic and<br />
international markets. However adoption seems far fetch from the reality and challenges to<br />
implementation across Africa will be discussed.<br />
Keywords: Bactrocera invadens, invasion, spread, bioecology, suppression.
PATAGONIA ARGENTINA. FRUIT FLY FREE AREA.<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Rial, Esteban J.*; Borges, Carlos A. & Mongabure, Alejandro P.<br />
Ingenieros Agrónomos. Fundación Barrera Zoofitosanitaria Patagónica. FunBaPa, Maipú 1634 General Roca<br />
(8332) Río Negro, Argentina. Email: erial@funbapa.org.ar.<br />
Background: The Patagonian Protected Region is a vast area of 835.000 km 2 and about 1.7<br />
million inhabitants. There are 150.000 hectares of irrigated valleys which form a real oasis<br />
within the typical semiarid plain landscape. These valleys are suitable for the production of<br />
vegetables, pome, stone and fine fruits (berries), all of which are in very good condition for<br />
export. The Region produces 1,000,000 tons of apples, 600,000 of pears (which represent the<br />
85% of the country´s production) and 100,000 tons of stone and fine fruits.<br />
Methods: The whole Region is naturally isolated by the Andean Range to the west, the Atlantic<br />
Ocean to the east and the Colorado and Barrancas Rivers, to the north. In addition to this, 13<br />
terrestrial inspection posts, 13 airports, 1 railroad control post and all the seaports of the<br />
Patagonian coast protect the area in order to prevent from any eventual re-infestation from the<br />
northern part of the country.<br />
Because of the presence of medfly in some urban areas, both Private and Public sectors in<br />
Patagonia decided, in 1997, to face the eradication of the pest in order to prevent quarantine<br />
restrictions imposed by some countries. After many years of hard work, combining regulation<br />
of incoming articles, Sterile Insect Technique, and cultural and chemical control, Patagonia<br />
was finally recognized as free of Fruit Flies by USDA/APHIS, by December 2005.<br />
The Program runs a 2.500 traps network, serviced every week, which demands 30 people<br />
working as trappers and samplers and identification laboratories. Control activities are<br />
concentrated, at present, on the implementation of the SIT through a Preventive Release<br />
Program over some urban areas where chances of a re-infestation are higher.<br />
Results: In case an outbreak occurs, an Emergency Plan, including focused control and<br />
regulatory activities, is immediately put into action. Since the eradication of the pest, two<br />
outbreaks occurred, one in 2006 and the other one in 2009. Both have been satisfied controlled<br />
and finally eradicated.<br />
Conclusions: This situation enabled Patagonia export to the United States for the fifth<br />
consecutive season without quarantine treatments.<br />
97
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ECONOMIC EVALUATION OF THE NATIONAL CAMPAIGN AGAINST NATIVE FRUIT<br />
FLIES IN SIX MEXICAN STATES (1994-2008).<br />
Salcedo Baca, Diznarda 1 *; Lomeli Flores, J. Refugio 1 ; Terrazas González, Gerardo H. 1 ; Rodríguez Leyva,<br />
Esteban & Enkerlin, Walther 2<br />
1 External consultants from the Interamerican Institute of Cooperation in Agriculture (IICA). Magnolias 20-B,<br />
Depto. 102, Col. Tlacoquemecatl del Valle, 03200 México, D.F., México; E-mail: diznarda.salcedo@cefprofit.com;<br />
2 Co-Director, Moscamed Regional Program.<br />
Background: The National Campaign against native fruit flies (Anastrepha spp.) was<br />
established by the Mexican Government in 1992 to prevent the negative effects of this pest in<br />
fruit loss in the field and quality of products, and through it, facilitate farmers the<br />
commercialization of their fresh fruits both in national and international markets. Thanks to the<br />
Campaign, several Mexican states and regions have been recognized as free areas (Baja<br />
California, Sonora, North & Central Sinaloa, Chihuahua and North of Zacatecas) and low<br />
prevalence areas of native fruit flies (South of Sinaloa, Nuevo León, Tamaulipas &<br />
Aguascalientes). This has facilitated observed increments in production and exports of<br />
Mexican fruits in the states where the Campaign has been operating. The different species of<br />
native fruit flies can cause up to 20% fruit loss in the field of sweet citrus and mango, 15% of<br />
guava and 30% of “mamey” and “chico zapote” if there were no phytosanitary measures<br />
implemented against this pest.<br />
Methods: Economic impacts were estimated through a retrospective model that was designed<br />
using as a guideline the “Cost-Benefit Analysis Model: A Tool for Area-Wide Fruit Fly<br />
Management” developed by FAO/IAEA. The designed model considered historical data of the<br />
observed variables, program costs and benefits (direct & indirect) for a 15 years period, to<br />
estimate the economic indicators: benefit-cost ratio (B/C), net present value (NPV), internal<br />
rate of return (IRR) and pay-back period.<br />
Results: Estimated economic indicators for each of the six evaluated states showed a B/C ratio<br />
much higher than one, positive NPV, very significant IRR and a pay-back period in only one<br />
year. A total investment in the Campaign of US $188 million for all six states during a 15 year<br />
period (6 for the state of Guerrero), generated direct and indirect benefits of US $1.8 and 1.5<br />
billion, respectively. Benefits for each state were: US $13 and 12 million, respectively, for Baja<br />
California, US $303 and 143 million for Guerrero, US $208 and 424 million for Nuevo León,<br />
US $578 and 340 million for Sinaloa, US $380 and 158 million for Sonora and US $347 and<br />
456 for Tamaulipas. For the mango produced in four of these states, direct benefits rose to US<br />
$960 million and indirect ones to US $548 million, while for the sweet citrus species cultivated<br />
in the six states, US $1.3 and 1.0 billion, respectively.<br />
Conclusions: Based on the obtained economic indicators, the decision of the Mexican<br />
government of establishing the National Campaign against Native Fruit Flies was rational from<br />
the economic point of view, since it has been significantly profitable to the six evaluated<br />
Mexican states during the analyzed period. Through this Campaign, Mexico has benefited<br />
from a growing production and exports of fruit, maintained employment in the agricultural<br />
sector and saved negative impacts to the environment. Keeping Mexico free of the<br />
Mediterranean fruit fly (a non native species) has been a strategic objective of the Mexican<br />
Government and a key in allowing the economic success of the National Campaign against<br />
Native Fruit Flies.<br />
Keywords: Native fruit flies, economic indicators, benefit to cost ratio
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
NEW TECHNOLOGY ON STERILE INSECT TECHNIQUE FOR FRUIT FLIES ECLOSION<br />
AND RELEASE IN MEXICO.<br />
Gutierrez, J. Manuel 1 ; Villaseñor, Antonio 1* ; Zavala, J. Luis 2* ; de los Santos, Martín 1 ;<br />
Leal, Rubén 3 & Alvarado, Ricardo 4<br />
1 Programa Moscas de la Fruta DGSV-SENASICA-México; 2 Programa Operativo Moscamed SENASICA-IICA<br />
México; Email: joseluiszavalalopez@yahoo.com.mx; 3 Servicios Aéreos, Biológicos y Forestales “Mubarqui”;<br />
4 Better World Manufacturing Inc.<br />
Background: The National Service for Plant, Animal and Forest Health of Mexico<br />
(SENASICA) have invested in building new facilities for packing, eclosion and aerial release<br />
of chilled adults of fruit flies to improve the Sterile Insect Technique (SIT).<br />
Methods: The technicians of Fruit Flies Program with the support of private enterprises have<br />
developed and improved new containers for packing and eclosion pupae and adults, towers<br />
type called “México” and “V&Z”, build with screened levels for lateral ventilation. With<br />
plastic accessories for pupae eclosion and to keep the puparia and residues for a clean free of<br />
paper process. A kind of plastic accessory increases surface and capacity to emerge a larger<br />
amount of pupae-flies as an easier alternative to “Worley” towers and PARC boxes containers.<br />
For feeding and drinking water, fruit flies in those new containers use a fruit flies balanced<br />
diet, 90% sugar and carbohydrates and 10% vegetable proteins, coming from a mixed of<br />
amaranth, peanut, egg powder, corn and sugar. Water is provided in a sponge or a cotton<br />
padded fabric. This is an alternative to the food made of Agar, sugar and water.<br />
For aerial release automatic machines are used with intelligent refrigerated boxes to keep<br />
temperature between 0-5 o C and humidity below 50%, for an optimal chilled adult flies release<br />
process. Those machines are provided with a conveyor (50cm. wide) working in transversal<br />
way to the axis of the plain. The released fly density per hectare is regulated by the speed of the<br />
conveyor and size of the gate of airplane chute. The precision release is made by aerial<br />
navigation system AGNAV, GPS. All process are recorded and monitored by an interactive<br />
Web site.<br />
Results and Conclusions: The new technologies for SIT have been applied since 2000 and have<br />
been released around 104,000 million of sterile flies of the Anastrepha genus and 61,200<br />
million of Ceratitis capitata (Wiedemann). All this have contributed to achieve the goals of the<br />
Fruit Fly Programs of Mexico, with the creation and expansion of free and low prevalence<br />
areas around 49% of Mexican territory and in the containment and eradication of Medfly in<br />
Mexico and Guatemala border.<br />
Keywords: Medfly, fruitfly, packing and eclosion, balanced diet , aerial release,<br />
99
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
NATIONAL FRUIT FLY CONTROL AND ERADICATION PROGRAM (PROCEM)<br />
ARGENTINA.<br />
Quiroga, Diego; Ramírez, Wilda* & Ruiz, Cynthia<br />
Servicio Nacional de Sanidad y Calidad Agroalimentaria (SENASA) – Dirección Nacional de Protección Vegetal<br />
– Av. Paseo Colón 315 (1063) 4° B – Capital federal – Buenos Aires – República Argentina. E-mail:<br />
dquiroga@senasa.gov.ar<br />
Background: The National Fruit Fly Control and Eradication Program was created in Argentina<br />
in 1994, with the aim to suppress and eradicate the species Ceratitis capitata (Medfly) and<br />
Anastrepha fraterculus (South American Fruit Fly), according to the agrobiological conditions<br />
of each region. Considering that the production of fruits and vegetables accounts for 20% of<br />
total Agricultural Gross Domestic Product (pome and stone fruits: 126,952 ha; citrus fruit –<br />
excluding lemon–: 96,965 ha; grapes: 226,450 ha), the main objectives of the Program are to<br />
reduce economic losses caused by fruit flies and to facilitate the access of fruit and vegetable<br />
products to international markets.<br />
Methods: The Program is currently implemented in 910,000 hectares, with the support of 1,060<br />
professionals and technicians, developing the following activities: Surveillance (setting of traps<br />
and fruit sampling to monitor Anastrepha fraterculus and Ceratitis capitata, and to detect<br />
exotic fruit flies, like: Ceratitis spp., Anastrepha spp., Toxotrypana curvicauda, Bactrocera<br />
spp., Rhagoletis spp. y Dacus spp.), Control (sterile insect technique, chemical control<br />
methods, biological control activities, cultural practices and legal control), SIT production<br />
(there are two rearing facilities, located in the Provinces of Mendoza and San Juan, which<br />
provide the sterile material to the areas under official program), Quarantine Protection System<br />
(phytosanitary barriers inspection and control vehicles and loads, 24 hours a day, all the year,<br />
using organic material detectors and/or detector dogs), Quarantine Treatment Facilities<br />
(approved quarantine treatments are applied to fruit fly host products, prior to their entry into<br />
the protected areas), Quality Controls, Training, Diffusion and Communication.<br />
Results & Conclusions: The program has achieved different status of plague, including: Fruit<br />
Fly–Free Areas (Central and Southern Oases of Mendoza Province and the Patagonia Region),<br />
Areas with Low Prevalence of C. capitata and Free of A. fraterculus (Northern and Eastern<br />
Oases of Mendoza Province), Areas with Low Prevalence of C. capitata and Under Control of<br />
A. fraterculus (Calingasta’s Valley of San Juan Province), Areas Under Control of C. capitata<br />
y A. fraterculus (Cultivated Valleys of San Juan Province) and Uncontrolled Population subject<br />
to Monitoring Surveys (Monte Caseros–Colón, Provinces of Corrientes and Entre Ríos).
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
STUDIES ON SMALL MODEL AREA-WIDE CONTROL OF THE ORIENTAL FRUIT FLY<br />
FOR WAX APPLE IN TAIWAN.<br />
Chiang, Ming-Yao*; Kao, Ching-Hua & Huang, Yu-Bin<br />
Applied Zoology Division, Taiwan Agricultural Reserach Institute (TARI), Wufeng, Taichung, Taiwan, ROC.<br />
Email: mingyaw@tari.gov.tw<br />
Background: The area-wide (AW) control of the oriental fruit fly (Bactrocera dorsalis<br />
(Hendel), OFF) was tested, using the conditions defined by Lindquist (2000), in Juan-Nan area<br />
of Mei-Shan township, Chia-Yi county, where wax apple is the major crop with significant<br />
economical importance. Growers in this area show a common interest in wax apple production,<br />
hence are highly cooperative in managing the fruit fly population.<br />
Methods: All available OFF control techniques have been utilized, e.g., methyl eugenol bait for<br />
male annihilation; protein bait for female fly control; bagging of the fruits; and thorough field<br />
sanitation in all wax apple orchards. A well organized OFF density monitoring program was<br />
established with 12 monitoring sites, which covered 500 ha testing area. By using 3D aerophotos<br />
display system, control planning and farmer education were conducted under the AW<br />
control scheme.<br />
Results: Comparing OFF density from individual monitoring sites in 2005 and 2006, the high<br />
density sites almost the same to conjecture that sites are hot spot possibility. The similar result<br />
reappears next year. In two years of AW control practice, the OFF population dropped below<br />
the economic threshold and fruit production was in excellent quality with a financial gain of<br />
NT$ 4 million ($ 122,000) in 2005. In 2006, the OFF population density was further reduced<br />
50% from that of 2005, and the estimated economic gain on fruit value combining with the<br />
saving of pest control cost exceeded NT$ 9 million ($ 286,000). The evaluation of the AW<br />
control program also included a survey of farmers’ inputs, which concluded a score of 4.0 in<br />
the scale of 1 to 5. The control practice covering 500 ha was considered small in the AW<br />
control model for OFF, as if did not have any buffer zone established. Despite the high<br />
mobility of OFF, no serious re-invasion problem from the adjacent area occurred due to the<br />
scarcity of alternative hosts in that area.<br />
Conclusions: This study, though confirmed the AW control requirements as proposed by<br />
Lindquist, recommends the requirements supplemented with such additional elements as<br />
effective control tactics, continuous farmer education, new and available technologies.<br />
Keywords: Bactrocera dorsalis, Area-wide control, Wax apple, Small acreage model.<br />
101
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DEVELOPING, IMPLEMENTING AND ECONOMIC ANALYSIS OF AREA-WIDE<br />
INTEGRATED MANAGEMENT OF MANGO FRUIT FLY, BACTROCERA DORSALIS<br />
(HENDEL) IN SOUTH INDIA<br />
Verghese, Abraham 1 & Mumford, John M. 2<br />
1 Indian Institute of Horticultural Research, Hesserghatta Lake PO, Bangalore 560089, India; Email:<br />
abraham.avergis@gmail.com; 2 Imperial College London, Silwood Park, Ascot, Berkshire SL 5 7PY, UK.<br />
Background: The fruit fly, Bactrocera dorsalis (Hendel) is a major pest of mango (Mangifera<br />
indica) causing on an average 27% loss in India. Additionally it has been an impediment in<br />
exports. So, an Integrated Management of Fruit Flies (IMFFI) project was initiated in India in<br />
2001, as a joint collaboration between Indian Council of Agricultural Research (ICAR, New<br />
Delhi) and Department for International Development (DFID, UK). The main objective was to<br />
develop an integrated management (IM) of the mango fruit fly B. dorsalis.<br />
Methods: The IM was developed and standardized, between 2001 and 2005. Between 2006 and<br />
2009, the IM was transferred to mango growers on an area-wide basis. The strategies<br />
investigated were male annihilation using methyl eugenol, field sanitation, bait application,<br />
visual (colour lures) and use of Oscimum extract. The economic impact of the IM was assessed<br />
using a semi-structured questionnaire in a major mango belt in South India.<br />
Results: It was found that methyl eugenol impregnated plywood (with dichorovos as toxicant)<br />
placed in a plastic container (500ml capacity) with two circular vents (1.5cm diameter) served<br />
as an ideal cost-effective trap [Cost per trap= 0.5 Euro]. It was worked out that 6-8 traps/acre<br />
could serve both as a surveillance tool and male annihilator (MA). A series of experiments with<br />
several indigenous baits showed that jaggery or banana pulp at 10% (mixed with a toxicant like<br />
malathion) was on par with protein hydrosylate, as splashes and could be used in IM as bait<br />
application technology (BAT). The number of BAT splashes were standardized to 40/acre, at<br />
the rate of 50-100ml bait solution per tree trunk (given two feet above the ground). Likewise,<br />
cultural approach of regular sanitation (weekly removal and destruction of fallen fruits) showed<br />
additive control. These measures, viz, MA, BAT and sanitation on an area-wide basis gave<br />
excellent control (95-100%). These areas were in pockets within a contiguous mango belt in<br />
south India between 12 0 E, 79 0 N. The impact analysis showed that these farmers realized at<br />
least 20-40% yield increase, with a cost: benefit ratio ranging from 1:4 to 1:20, depending on<br />
the commercial value of the mango variety. Visual and Oscimum lures were not effective.<br />
Conclusions: Farmers expressed high satisfaction with the area wide IM of fruit flies. So it is<br />
envisaged, to cover IM of fruit fly on a wider area, in 2010 and 2011 with financial support<br />
from ICAR and NABARD (National Bank for Agricultural and Rural Development, India.).<br />
This will further strengthen the adoption of fruit fly IM in mango in India and South Asia<br />
where Bactrocera spp are a problem.<br />
Keywords: Bactrocera spp, Mango, area-wide, male annihilation, baits, integrated management, India
Session 7<br />
Natural Enemies & Biocontrol
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
INSECTICIDAL COMPOUNDS SECRETED BY ENTOMOPATHOGENIC FUNGI SHOW<br />
PROMISE FOR CERATITIS CAPITATA (WIEDEMANN) CONTROL.<br />
Quesada-Moraga, Enrique*; Santiago-Álvarez, Cándido & Ortiz-Urquiza, Almudena<br />
Agricultural Entomology Unit. Department of Agricultural and Forestry Sciences, University of Cordoba. Campus<br />
de Rabanales C4, 2º planta 14071 Córdoba, Spain. Email: cr2qumoe@uco.es.<br />
Background: The control of Ceratitis capitata is usually performed with protein bait sprays<br />
incorporating chemical insecticides that have adverse effects on humans, non-target organisms,<br />
and environment. For that, in the last decade, there has been an increasing interest for the so<br />
known natural insecticides for medfly control, including those from higher plants and microorganisms.<br />
Among microorganism, naturally occurring entomopathogenic fungi (EF) not only<br />
are excellent candidates for medfly microbial control but also a new poorly explored source of<br />
novel insecticidal compounds of natural origin<br />
Methods: The crude soluble protein extract (CSPE) in liquid media of 130 isolates from our<br />
collection of autochthonous strains of EF obtained from the soil, insect and phyllo plane<br />
belonging to several EF species, with emphasis in Metarhizum anisopliae, Beauveria bassiana,<br />
have been evaluated for insecticidal activity against newly emerged medfly adults when<br />
administered per os. The active fractions of the CSPE have been purified by liquid<br />
chromatography and gel electro-elution. The active protein fractions have been characterized<br />
and their acute, chronic and sublethal reproductive effects on medly adults evaluated.<br />
Results: To now, only three out of the 130 CSPEs evaluated have shown insecticidal activity<br />
against medfly adults. Among them, the CSPE of M. anisopliae EAMa 01/58-Su strain is the<br />
most active. The mortality in flies fed with this CSPE exhibits a dose and time-related<br />
response, a result likely due to a progressive deterioration on the fly midgut after ingestion of<br />
the extract. Four monomeric proteins from this crude extract have been purified by liquid<br />
chromatography and gel electro-elution. Although all four monomers seem to be involved in<br />
the insecticidal activity of the CSPE, the 15 kDa and the 11 KDa proteins appear largely<br />
responsible for the observed insecticidal effect. Besides, the activity of the CSPE of this isolate<br />
can be highly enhanced (by seven fold) by manipulating the nutrient conditions of the liquid<br />
medium (carbon and nitrogen sources and ratios). Likewise, a reduction of female fecundity<br />
levels during the first days after the treatment has been found in CSPE challenged flies.<br />
Conclusions: We have presented evidence that the CSPE of M. anisopliae 01/58-Su strain<br />
should be considered as a new tool for biocontrol of C. capitata adults that can be combined<br />
with an attraction bait-system to increase its efficiency while minimizing the impact on<br />
beneficial insects.<br />
Keywords: natural insecticides, medfly, fungal proteins, metabolites, Beauveria, Metarhizium, baits, fungal<br />
extracts, macromolecules, toxicity<br />
105
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CERATITIS CAPITATA (WIEDEMANN) SUSCEPTIBILITY TO CYT1AA TOXIN FROM<br />
BACILLUS THURINGIENSIS (BERLINER).<br />
Vidal-Quist, J. Cristian 1* ; Castañera, Pedro 2 & González-Cabrera, Joel 2<br />
Unidad Asociada de Entomología UJI-IVIA-CIB CSIC. 1 Instituto Valenciano de Investigaciones Agrarias (IVIA).<br />
Centro de Protección Vegetal y Biotecnología; Ctra. Moncada-Náquera km 4.5; E-46113-Moncada. Spain. e-mail:<br />
vidal_josqui@gva.es; 2 Centro de Investigaciones Biológicas (CIB CSIC). Madrid, Spain.<br />
Background: The Mediterranean fruit fly, Ceratitis capitata, is one of the major threats to fruits<br />
crops worldwide. Current control methods rely mostly on synthetic insecticides. Their impact<br />
on human health and the environment along with the development of resistances make<br />
necessary the implementation of sustainable control methods. Bacillus thuringiensis (Bt)<br />
based-products should be a good alternative since they have been used for decades for<br />
controlling economically important pests. Though δ-endotoxins are highly toxic and specific<br />
against many insect pests, including dipterans, previous works performed in our group have<br />
reported that C. capitata is not susceptible to spore and crystal (S+C) mixtures (containing δendotoxins)<br />
obtained from a wide array of Bt isolates. The goal of this work was to overcome<br />
the reported inefficiency of Bt S+C mixtures against C. capitata by the administration of<br />
solubilised and/or activated δ-endotoxins.<br />
Methods: Our first approach was to test the in vitro solubilized crystal protoxins obtained from<br />
cultures of 35 native selected strains and 5 standard strains. Proteolytic activation of δendotoxins<br />
produced by the most toxic strains was also tested; the crystals were in vitro<br />
solubilised and thereafter incubated with gut extracts from 3 insect species (C. capitata,<br />
Sesamia nonagrioides and Culex pipiens).<br />
Results: Bioassays on neonate C. capitata larvae showed significant sublethal effects of<br />
solubilised protoxins at 20 µg/cm 2 on one standard and two native strains, all belonging to the<br />
Bt serotype israelensis (Bti). Protein electrophoresis revealed that protoxin activation was<br />
strongly dependent on the source of proteases. Dose-response assays showed that in vitro<br />
proteolytic processing of Bti protoxins increased lethal effects on C. capitata neonate larvae,<br />
being significantly higher when protoxins were activated with Culex pipiens proteases, a<br />
dipteran species highly susceptible to Bti. LC50 of C. pipiens activated Bti toxins was 31.26<br />
µg/cm 2 . Additionally, the use of a recombinant Bt strain allowed the identification a single δendotoxin,<br />
Cyt1Aa, responsible for the lethality found on C. capitata larvae. LC50 of this<br />
protoxin after solubilisation was: 4.93 µg/cm 2 .<br />
Conclusions: We have shown that in vitro emulation of events from Bt mechanism of action<br />
may increase the activity against recalcitrant pests such as C. capitata. The identification of a<br />
single δ-endotoxin with larvicidal activity may be the basis for new engineered Bt strains<br />
effective on C. capitata control.<br />
Keywords: Ceratitis capitata, Sesamia nonagrioides, Culex pipiens, proteases, bioassays
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
COMPATIBILITY OF A NEW ATTRACTANT-CONTAMINANT DEVICE CONTAINING<br />
METARHIZIUM ANISOPLIAE WITH MEDFLY SIT PROGRAMS.<br />
San Andrés, Victoria 1,2 ; Moya, Pilar 3* ; Ayala, Ildefonso 3 ; Abad, María 3 ; Primo, Jaime 1 &<br />
Castañera, Pedro 2<br />
1 Instituto Valenciano de Investigaciones Agrarias (IVIA), Unidad Asociada de Entomología IVIA– CIB CSIC.<br />
Ctra. de Moncada a Náquera km. 4,5; Moncada 46113- Valencia (Spain); 2 Centro de Investigaciones Biológicas<br />
(CIB) del Consejo Superior de Investigaciones Científicas (CSIC), Departamento Biología de Plantas, Madrid,<br />
Spain; 3 Centro de Ecología Química Agrícola (CEQA), Universidad Politécnica de Valencia (<strong>UPV</strong>), Camino de<br />
Vera s/n. 46022. Valencia, Spain. Email: mmoyasa@ceqa.upv.es.<br />
Background: The Sterile Insect Technique (SIT) is one of the most useful strategies to control<br />
the Mediterranean fruit fly Ceratitis capitata (Wiedemann), medfly. However, new or<br />
improved eco-friendly control methods are required to synergise or complement SIT programs.<br />
One of the new tools currently being studied is the use of entomopathogenic fungi. Recently, a<br />
new C. capitata attractant-contaminant device, containing Metarhizium anisopliae<br />
(Metschnikoff) Sorokin, developed by our group, showed a high field efficacy on this citrus<br />
pest. The aim of this work was to evaluate the compatibility of ongoing SIT programs in the<br />
Comunidad Valenciana with this “attractant -contaminant” technique. The combination of<br />
these two techniques would be feasible providing that at least the sterile males do not show a<br />
higher susceptibility to M. anisopliae than the wild population, and that the mating<br />
performance of the sterile male is not affected.<br />
Methods: Susceptibility of C. capitata Vienna 8 males to M. anisopliae was evaluated by<br />
topical application and compared with that previously obtained from a C. capitata laboratory<br />
strain, annually refreshed with wild medflies (WTL strain). A comparative mating performance<br />
bioassay between sterile and WTL medfly males exposed to attractant-contaminant devices<br />
(fungus contaminated or fungus-free) was also carried out. One day after the exposure, healthy<br />
virgin WTL females were allowed to mate with each male group. Mating pairs were<br />
individually collected during three hours and the number of mating and their duration recorded.<br />
Mated medfly females were kept in separated cages for two days, and then allowed to remate<br />
with non-treated WTL males. Then, remated females were maintained individually in separated<br />
cages to determine the fecundity and fertility.<br />
Results: The pathogenicity bioassays established that the susceptibility of C capitata Vienna 8<br />
males to our M. anisopliae strain was similar to that showed by the WTL strain, and did not<br />
impair the percentage of mating. However, the duration of the copula increased significantly<br />
for copulating WTL male treated with M. anisopliae. No significant differences on fecundity<br />
were found between females mated with the M. anisopliae treated and not treated males, but<br />
the fertility of remated females was significantly higher for those mated twice with males of<br />
the WTL strain, despite of the treatment.<br />
Conclusions: M. anisopliae does not adversely affect any of studied parameters linked to<br />
medfly mating performance, suggesting that the “attractant-contaminant” technique would be<br />
compatible with SIT programmes.<br />
Keywords: Medfly, entomopathogenic fungi, biological control<br />
107
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PEST, PARASITOID AND FRUIT INTERACTIONS IN BIOLOGICAL CONTROL OF THE<br />
OLIVE FRUIT FLY IN CALIFORNIA.<br />
Yokoyama, Victoria Y.*<br />
U.S. Department of Agriculture, Agricultural Research Service, San Joaquin Valley Agricultural Sciences Center<br />
(USDA-ARS-SJVASC), 9611 South Riverbend Avenue, Parlier, California 93648 USA. Email:<br />
Victoria.Yokoyama@ars.usda.gov<br />
Background: Olive fruit fly, Bactrocera oleae, has become a key pest in olives, Olea europaea,<br />
since it was first discovered in California in 1998. Biological control is an economical<br />
alternative for costly bait sprays in commercial orchards of olives grown for canning and oil.<br />
Several years of augmented releases of an imported parasitoid, Psyttalia humilis, has resulted<br />
in successful parasitism of olive fruit fly, but the parasitoid has not yet become established.<br />
Methods: The life cycle of olive fruit fly and P. humilis was studied in the laboratory,<br />
greenhouse, and field. Survival of the adult and immature stages of olive fruit fly and the adult<br />
stage of P. humilis was investigated at constant temperatures (15-25°C) and humidities (35-<br />
65%) in laboratory incubator tests, and in two greenhouse climates with fluctuating diurnal<br />
(32-36°C, 31-37% hot vs. 26°C, 61-63% cool) and nocturnal conditions, and with and without<br />
honey for food and water. Seasonal field temperatures and humidities were monitored with<br />
data loggers. Olive fruit fly adult infestations were determined in the field with yellow sticky<br />
traps baited with ammonium carbonate baits and pheromone lures. Fruit samples were<br />
collected to determine natural larval infestations and parasitism after parasitoid releases.<br />
Results: Olive fruit fly and P. humilis required cool temperatures, high humidities, and food<br />
and water for prolonged survival (≈6 months for host) in laboratory and greenhouse tests. Life<br />
span was greatly shortened by high temperatures, low humidities, and the absence of food and<br />
water. The observations were consistent with high populations of the pest found in cool coastal<br />
areas and low numbers that occur in dry, arid inland valleys. The parasitoid was effective for<br />
biological control (≤100% parasitism) when fruit infestations were high such as in coastal olive<br />
growing areas. Non-establishment of the parasitoid was related to the lack of infested olive<br />
fruit remaining in the trees during winter. The major factors that caused infested fruit suitable<br />
for development of P. humilis to drop from trees were high larval infestations and occasional<br />
freezing winter conditions. The parasitoid could survive (≈4 months at 15°C) and reproduce<br />
(10°C) in the host during winter. Olive fruit fly larvae were found to complete development in<br />
immature fruit 1 cm long increasing availability of host between annual crops.<br />
Conclusions: Successful biological control of olive fruit fly in California is dependent on the<br />
presence of infested fruit which is destroyed by high larval numbers and inclement weather.<br />
The parasitoid is long-lived and capable of overwintering as adults, but the host may be<br />
unavailable due to fruit condition.<br />
Keywords: Olive fruit fly, Psyttalia humilis, biological control, olives
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CONSERVATION BIOLOGICAL CONTROL STRATEGIES IN CITRUS AS A TOOL TO<br />
REGULATE MEDFLY POPULATIONS.<br />
Monzó, César* 1,2 ; Urbaneja, Alberto 1 ; Sabater-Muñoz, Beatriz 1 & Castañera, Pedro 2<br />
1 Instituto Valenciano de Investigaciones Agrarias (IVIA), Unidad Asociada de Entomología IVIA– CIB CSIC.<br />
Ctra. de Moncada a Náquera km. 4,5, Moncada 46113- Valencia (Spain); Email: cmonzo@ivia.es; 2 Centro de<br />
Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC), Departamento<br />
Biología de Plantas, Madrid, Spain.<br />
Background: In recent years, emphasis has been placed on implementing environmentally<br />
friendly methods to control medfly. Within this context, biological control measures based on<br />
predators have hardly been used. In medfly there are three developmental stages that are<br />
susceptible to being preyed upon by ground-dwelling predators: late third instars larvae, which<br />
jump from the fruit to pupate underground, pupae and teneral adults, which remain on the soil<br />
until they are able to fly. We report here on the importance of these ground-dwelling predators<br />
in citrus groves and their influence upon C. capitata.<br />
Methods: A 3-years study of the ground-dwelling predator assemblages associated to citrus<br />
orchards has been conducted in Valencia Region (Spain) using pit-fall traps. Activity-density<br />
patterns, community structure, species richness and abundance parameters of these<br />
assemblages were evaluated. Functional responses against the medfly developmental stages<br />
susceptible of being preyed upon were obtained under laboratory conditions for the prevalent<br />
predator species of each group. A C. capitata specific PCR marker was designed and used to<br />
assess medfly predation by means of predator gut-content analysis.<br />
Results: This work reveals the presence of a rich and abundant complex of ground-dwelling<br />
polyphagous predators inhabiting the ground surface of citrus orchards throughout the year.<br />
Indeed, 17,526 ground-dwelling predator specimens belonging to 110 different species were<br />
captured. The prevalent predators found were the lycosid Pardosa cribata (Araenae), the<br />
ground beetle Pseudophonus rufipes (Coleoptera: Carabidae) and the earwig Forficula<br />
auricularia. All these prevalent predators were able to prey upon C. capitata under laboratory<br />
conditions. Pseudoophonus rufipes was the most efficient predator, while F. auricularia was<br />
the least. Pseudoophonus rufipes preyed mainly upon pupae, with an estimated attack rate of<br />
3.07 d -1 and a handling time of 0.048 d, P. cribata used teneral adults as the main prey, with an<br />
estimate attack rate of 0.771 d -1 and a handling time of 0.051 d, and F. auricularia showed the<br />
highest preference for third-instar larvae, with an estimated attack rate of 0.269 d -1 and a<br />
handling time of 0.065 d. The carabid P. rufipes turned out to be the most efficient predator<br />
under field conditions, with 23% specimens testing positive after PCR gut-content analysis,<br />
while the proportion of P. cribata testing positive was 5%.<br />
Conclusions: Ground-dwelling predators could play an important role on regulating C. capitata<br />
populations. A challenge for the future will be how to enhance their populations, to increase<br />
their efficiency against this pest, and consequently, to incorporate them in current citrus<br />
conservation biological control strategies.<br />
Keywords: Ground-dwelling polyphagous pedators, medfly, functional response, PCR gut-content analysis,<br />
conservation biological control.<br />
109
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
A NEW OVIPOSITION-DETERRING ANT CUE PROTECTS MANGOES FROM FRUIT<br />
FLIES.<br />
Vayssières, Jean-François* 1 ; Sinzogan, Antonio 1 ; Adandonon, Appolinaire 1 ;<br />
Ouagoussounon, Issa 1 & Van Mele, Paul 2<br />
1 International Institute of Tropical Agriculture, Centre International de Recherche (IITA-CIRAD), 08 BP 0932,<br />
Cotonou, Benin. Email: j.vayssieres@cgiar.org; 2 Africa Rice Center (ARC), Cotonou, Benin.<br />
Background: The mango fruit industry in sub-Saharan Africa suffers quality losses from fruit<br />
flies. The impact of African weaver ant (Oecophylla longinoda) on fruit fly damage to earlier<br />
mango cultivars (cv) was studied, along with the tritrophic interactions between mangoes<br />
(Mangifera indica), two frugivorous fly species (Bactrocera invadens and Ceratitis cosyra)<br />
and weaver ants. These observations showed that olfactory and visual factors from the presence<br />
of O. longinoda could help deter these pests. We investigated ant impact on season and late<br />
cultivars and the extent to which the density of the pheromone of ants affects the oviposition<br />
behaviour of two major mango fruit fly species and resultant damage.<br />
Methods: In the field, we investigated weaver ant impact on three seasonal mango cv (Ifac 3,<br />
Améliorée du Cameroun, Kent) and two late cv (Smith and Brooks). We used three blocks in<br />
2008 (Ifac 3, Kent, Smith) and three other blocks in 2009 (Améliorée, Smith and Brooks), each<br />
with two treatments repeated 10 times: (i) without ants = control, (ii) with ants. In the lab, the<br />
effect of Oecophylla cues on B. invadens and C. cosyra oviposition behaviour were studied<br />
using ant-exposed and unexposed mangoes as treatments in 10 replicates. Individual fruits<br />
were placed in separate cages and confined with three gravid females of either B. invadens or<br />
C. cosyra.<br />
Results: (1) mean damage inflicted by fruit flies was significantly reduced by 58% and 81% in<br />
2008 and 2009, respectively; (2) tephritid species oviposited significantly less in fruits<br />
previously patrolled by ants; (3) for landing, C. cosyra seemed twice as sensitive to the ant<br />
cues than B. invadens on exposed fruits vs. unexposed; (4) although B. invadens generally<br />
takes longer than C. cosyra to oviposit, there was no significant difference between species<br />
when confined with ant-exposed fruit; (5) damage from B. invadens was six times lower and<br />
from C. cosyra four times lower for ant-exposed fruits vs. unexposed fruits;(6) B. invadens had<br />
significantly more pupae per kg of fruit than C. cosyra when confined with unexposed<br />
mangoes. No significant difference was detected between species when confined with antexposed<br />
fruits. We discuss the significance of prey recognition of cues from generalist<br />
predators and resulting avoidance, together with ecological and behavioural implications.<br />
Conclusions: The use of Oecophylla colonies is well-suited to perennial cropping systems in<br />
sub-Saharan Africa, as they are efficient, constantly available, widespread and selfregenerating.<br />
The effect of cues from generalist predators, such as ants, on the foraging<br />
behaviour of frugivorous insects and more generally on pests in terrestrial systems is a<br />
relatively new finding likely to have crucial consequences for future research on developing<br />
protection strategies against tephritid pests.<br />
Keywords: conservation biological control, predator avoidance, tritrophic relationship, Mangifera indica.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
REARING THE BIOLOGICAL CONTROL AGENT DIACHASMIMORPHA KRAUSSI<br />
(FULLAWAY) (HYMENOPTERA: BRACONIDAE) ON IRRADIATED LARVAE OF THE<br />
QUEENSLAND FRUIT FLY, BACTROCERA TRYONI (FROGGATT) (DIPTERA:<br />
TEPHRITIDAE).<br />
Reynolds, Olivia L. 1 *; Harris, A.R. 2 ; Pratt, C.F. 2 ; Jessup, Andrew J. 3 ; Banos, C. 4 ;<br />
Lindhout, K. 5 & Gurr, Geoff M. 6<br />
1 EH Graham Centre for Agricultural Innovation, Industry & Investment New South Wales, Private Bag 4008,<br />
Narellan NSW, Australia 2567; Email: olivia.reynolds@industry.nsw.gov.au; 2 Imperial College London, Division<br />
of Biology, Silwood Park, Ascot, Berkshire, SL5 7PY, United Kingdom; 3 Insect Pest Control Sub-programme,<br />
FAO/IAEA Agriculture and Biotechnology Laboratory, A-2444 Seibersdorf, Austria; 4 Australian Nuclear Science<br />
and Technology Organisation, Menai, NSW, 2234; 5 Industry & Investment New South Wales, Narara, NSW,<br />
Australia 2250; 6 EH Graham Centre for Agricultural Innovation, Charles Sturt University, Orange, NSW,<br />
Australia 2800.<br />
Background: The Queensland fruit fly, Bactrocera tryoni is one of the most damaging pests in<br />
Australia. Biological control would be a welcome technology. This paper presents results from<br />
a study which aimed to assess scope for mass rearing parasitoid wasps on gamma irradiated<br />
host larvae. The practical advantage of such a rearing system is that flies and wasps would not<br />
need to be separated, thus reducing cost of production. Wasp production could also be done<br />
without fear of adverse biological or political consequences arising from escapees.<br />
Methods: Diachasmimorpha kraussii adults of both sexes emerged from hosts that were<br />
irradiated and exposed as second instar larvae irrespective of dose (0, 4.7, 9.1, 15.9, 27.6, 47.0<br />
and 79.9 Gy).<br />
Results: The fitness of wasps appeared unaffected by the host irradiation; adults from all<br />
treatments produced equally numerous offspring. For hosts that were not parasitised, fertile<br />
adult B. tryoni of both sexes emerged from the control treatment and at the lowest two<br />
irradiation doses. Only two flies emerged from the 15.9 Gy treatment and both died within two<br />
days. High levels of sterility were not observed for B. tryoni at any dose. In similar work in<br />
which hosts were irradiated and exposed as third instar larvae, wasps developed regardless of<br />
dose. Adult B. tryoni of both sexes emerged from the control treatment and the 4.7 and 9.1 Gy<br />
doses for all replicates. Emergence declined as the irradiation dose increased and only one fly<br />
emerged from the 79.9 Gy dose and this died within a day.<br />
Conclusions: Overall results suggest scope to develop a protocol involving a carefully<br />
calibrated irradiation dose and host material of a specific developmental stage that will allow<br />
mass production of D. kraussii, yet either excludes fly emergence or renders flies sterile from<br />
any unparasitised hosts.<br />
Keywords: parasitoid, gamma irradiation, opine, natural enemy<br />
111
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF HOST REARING DIET ON FITNESS PARAMETERS OF FOPIUS ARISANUS<br />
(SONAN) (BRACONIDAE: HYMENOPTERA).<br />
Mohamed, Samira A. 1 *, Ekesi, Sunday 1 & Hanna, Rachid 2<br />
1 International Centre of Insect Physiology and Ecology, P.O. Box, 30772-00100, Nairobi, Kenya, Email:<br />
sfaris@icipe.org; 2 The International Institute of Tropical Agriculture (IITA), Yaounde, Cameroon.<br />
Background: Bactrocera invadens, an alien invasive pest in Africa poses a significant threat to<br />
the livelihoods of millions of African farmers and the entire African economy which is largely<br />
agricultural based. To combat the devastating effect of this aggressive pest an IPM package<br />
was developed and validated by icipe. Classical biological control using a coevolved<br />
parasitoid, Fopius arisanus (Braconidae: Hymenoptera) constitutes an essential component of<br />
this IPM package. However, to insure the releases of this parasitoid on a large scale in all<br />
invaded countries, a sustainable and affordable rearing media for mass production of good<br />
quality wasps is urgently needed. In this study the effect of three host rearing media; on various<br />
fitness parameters of Fopius arisanus was evaluated.<br />
Methods: Parasitized B. invadens eggs were reared on three media; liquid diet, carrot based diet<br />
and mango pulp. Number of recovered puparia, puparia weight, percentage uneclosed puparia,<br />
percentage of total emerged wasps, developmental time, potential fecundity, adult wasp’s size<br />
and adult’s longevity were recorded.<br />
Results: Total puparia recovery, puparia weight and percentage emerged wasps were<br />
significantly higher on liquid diet compared with other diets. Also wasps that emerged from<br />
hosts reared on liquid diet had shorter developmental time, larger in size, and their females<br />
were more fecund. Wasps’ longevity was comparable across the diets. However females lived<br />
longer than males in all diet types tested. Uneclosed puparia significantly varied with the diet<br />
types being lowest when the host was reared on mango but comparable for liquid and carrot<br />
diet.<br />
Conclusions: Based on the results of this study, F. arisanus wasps from the host reared on the<br />
liquid diet were of superior quality for almost all the tested parameters. The potential of<br />
adopting the use of liquid diet on a large scale and replacing some of its ingredients with less<br />
expensive and readily available local alternatives will be discussed.<br />
Keywords: Bactrocera invadens, Fopius arisanus, mass rearing, quality control
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FIELD SUPERPARASITISM OF DIACHASMIMORPHA LONGICAUDATA<br />
(HYMENOPTERA: BRACONIDAE) ATTACKING ANASTREPHA LARVA ON MANGO<br />
FRUITS.<br />
Montoya, Pablo 1 ; Ruiz, Lía 1 ; Cancino, Jorge 1 ; Pérez-Lachaud, Gabriela 2 & Liedo, Pablo 2<br />
1 Subdirección de Desarrollo de Métodos, Programa Moscafrut SAGARPA-IICA. Central Poniente 14, Col Centro,<br />
CP 30700, Tapachula, Chiapas, México. Email: pmontoya@prodigy.net.mx; 2 El Colegio de la Frontera Sur,<br />
Carretera Antiguo Aeropuerto Km 2.5, CP 30700, Tapachula, Chiapas, México<br />
Background: Diachasmimorpha longicaudata is one of the fruit flies larval parasitoids more<br />
used as augmentative biological control agent worldwide, given the relative ease of its mass<br />
rearing and the high efficiency of its releases in the field. Recent laboratory studies revealed<br />
that superparasitism in this species might have adaptive connotations; since it was found a<br />
significant correlation between the level of superparasitism and sex ratios biased to females,<br />
without detriment in other fitness parameters as fertility and longevity. Our aim in this work<br />
was to determine the presence of superparasitism in this species at field level, and study its<br />
possible relationship to variables such as host fruit size, size of parasitized larvae, and sex ratio<br />
of the emerged adults.<br />
Methods: We collected 498 mango fruits var. Creole (Manguifera indica) in the Valley of<br />
Mazapa de Madero, Chiapas, Mexico. We obtained 14,450 Anastrepha spp. larvae, which were<br />
placed in 1L containers with humid vermiculite to facilitate pupation. Once the pupae were<br />
formed, they were measured (length x diameter) with a digital micrometer, and weighed on an<br />
analytical balance. The presence of superparasitism was determined by counting the number of<br />
scars on each puparium, and sex ratio was determined at the emergence of adult parasitoids (n<br />
= 279).<br />
Results: We found that only 37.66% of the larvae were parasitized, and that 22% of them<br />
experienced superparasitism. The number of oviposition scars per pupa ranged between 2 and<br />
29 scars, although high levels were rare. The percentage of pupae with more than three scars<br />
was 8.11%. Fruit size showed no relationship to levels of parasitism and superparasitism, but it<br />
was found that the length and width of the pupae were positively related to superparasitism<br />
level (F1, 278 = 16.52, P = 0.0001, F1, 278 = 14.74, P = 0.0002, respectively). Through a logistic<br />
regression, we determined that a sex ratio biased to females was positively correlated with the<br />
number of scars per pupa and to the length of the pupa, but not with the width (� 2 1 for number<br />
of scars = 11.24, P = 0.0008; for pupal length = 4.303, P = 0.038; for pupa width = 1.157, P =<br />
0.21 NS). The adult emergence and a female biased sex ratio were favored when<br />
superparasitism levels were high (> 6 scars / pupa).<br />
Conclusions: Superparasitism is present in natural populations of D. longicaudata, and shows<br />
advantageous features such as increased adult parasitoid emergence and a female biased sex<br />
ratio. These findings are consistent with data found under mass rearing conditions, and support<br />
the election of this species as a viable candidate for biological control programs against fruit<br />
flies.<br />
Keywords: Parasitoids, Biological control, Fruit flies, sex ratio, oviposition scars.<br />
113
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ASEXUAL VERSUS SEXUAL STRAINS: WOLBACHIA INFECTION EFFECTS ON THE<br />
BIOLOGICAL CONTROL POTENTIAL OF ODONTOSEMA ANASTREPHAE BORGMEIER<br />
PARASITOIDS OF FRUIT FLIES.<br />
Davies, Andrew P. 1 ; Aluja, Martin 2 & Sivinski, John* 1<br />
1 United States Department of Agriculture – Agricultural Research Service (USDA-ARS), Center for Medical,<br />
Agricultural and Veterinary Entomology, Gainesville, Florida, USA. Email: John.Sivinski@ars.usda.gov; 2<br />
Instituto de Ecologia, A.C., Apartado Postal 63, 91000 Xalapa, Veracruz, Mexico.<br />
Background: The maternally inherited intracellular bacterium, Wolbachia pipientis, induces<br />
reproductive alterations in host insect populations such as male-killing, feminization,<br />
parthenogenesis induction (PI, or reproduction without males) and cytoplasmic incompatibility.<br />
Both thelytokous (asexual) and arrhenotokous (sexual) Odontosema anastrephae populations<br />
attack Anastrepha spp. fruit flies on guava in Mexico. Here we report that PI in Mexican O.<br />
anastrephae populations is caused by Wolbachia infection. In addition, we discuss implications<br />
derived from laboratory based bionomic comparisons of sexual and asexual O. anastrephae<br />
strains on their respective mass rearing and fruit fly biological control potentials.<br />
Methods: Wolbachia infection in field collected and laboratory reared O. anastrephae was<br />
detected and sequence typed using PCR. That Wolbachia is the likely cause of PI in asexual O.<br />
anastrephae was determined by PCR screening for alternative bacterial infections (both 16S<br />
rDNA general eubacterial primers and primers specific for other bacteria that cause PI) and<br />
antibiotic curing (0.5% tetracycline hydrochloride in 10% honey solution). For comparative<br />
bionomic studies, treatments of 10 asexual and mated sexual O. anastrephae females were<br />
each supplied with 10% honey solution as food and equal volumes of late instar Anastrepha<br />
suspensa larvae in diet as hosts. The experiment incorporated three replications and repetitions<br />
and A. suspensa larvae controls devoid of parasitoids. The numbers of emergent fruit flies,<br />
parasitoids and fruit fly pupae that did not eclose per exposure were counted.<br />
Results: Multi locus sequence typing determined that asexual O. anastrephae from Mexico are<br />
infected with a novel Wolbachia strain (ST 165). That no other bacterial infections were<br />
detected and exposure to antibiotics induced males suggests that Wolbachia causes PI in<br />
Mexican O. anastrephae populations. Asexual and sexual O. anastrephae sex ratios were<br />
1:64.7 and 1:2.5, respectively, and both strains displayed protandry. The proportion of<br />
emergent fruit flies (0.11:0.03) and percentage parasitism (0.33:0.20) in asexual treatments was<br />
higher than that in sexual treatments, while a higher proportion of fruit fly pupae demonstrated<br />
no eclosion in the latter (0.56:0.77).<br />
Conclusions: Wolbachia infection causes PI in O. anastrephae producing a higher proportion<br />
of females relative to uninfected sexual strains, yet potential infection benefits on mass<br />
production of the parasitoid remain unclear. Ongoing experiments aim to further elucidate<br />
strain differences and model their relative potential as biological control agents attacking fruit<br />
flies on guava in Mexico.<br />
Keywords: Parthenogenesis induction, endosymbionts, MLST, biological control
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
BACTEROCERA ZONATA (SAUNDERS) STATUS AND ITS NATURAL ENEMIES IN<br />
EGYPT.<br />
El-Heneidy, Ahmed H. * 1 & Ramadan, Mohsen M. 2<br />
1 Department of Biological Control (DBC), Plant Protection Research Institute (PPRI), Agricultural Research<br />
Centre (ARC), Giza, EGYPT. P.O. Box # 915, Postal Code 11728, Maadi, Cairo, EGYPT. E-mail:<br />
aheneidy@link.net; 2 State of Hawaii Department of Agriculture, Division of Plant Industry, Plant Pest Control<br />
Branch, Honolulu, HAWAII 96814 (USA)<br />
Background: The tephritid peach fruit fly (PFF), Bacterocera zonata is a recently introduced<br />
exotic fruit fly species that has established and become a widespread pest in Egypt. It was<br />
ranked as an A1 serious quarantine pest. B. zonata threatens countries in the Near East and<br />
North Africa and to a lesser extent in Southern Europe. PFF has not established yet in any other<br />
region of the Mediterranean or Africa so quarantine concerns are likely to be much more<br />
significant for Egypt. PFF poses a threat to many of Egypt’s export markets and a more<br />
immediate threat to the welfare of the industry than the Medfly and for this reason is likely to<br />
prevent further spread. PFF populations are building up slowly from early April reaching high<br />
levels in July which persist well in to the fall. Its major host plants in Egypt are; mango, guava,<br />
peach, apricot and citrus. PFF has been also recorded as a secondary pest species on some<br />
vegetable crops; cucurbits, tomatoes, paprika, eggplant and potatoes in Egypt. Such transaction<br />
of the pest from mainly fruit crops to some vegetable crops threats the production and export of<br />
those crops in the future in Egypt.<br />
Methods: PFF detection in Egypt depends on scattered traps using non-toxic methyl eugenol<br />
plugs. Control methodology, using killing bags, bait spraying of tree trunks beside chemical<br />
control has relatively limited effect on the pest control.<br />
Results: Through a wide survey, natural enemies, particularly native parasitoid species, seem to<br />
be absent. For nonnative pests, conservation is likely to be inadequate because sufficiently<br />
effective natural enemies will be absent. In such cases, introducing new effective natural<br />
enemy species as additional mortality factors against the pest is absolutely essential, and is an<br />
approach that historically has been extremely successful. Therefore, an US-Egypt collaborative<br />
project was proposed for utilizing the experience of the Plant Protection Dept., in Hawaii, USA<br />
in validating and implementing a biological control program for controlling the target fruit flies<br />
in Egypt, through testing some exotic specific parasitoid species, as well exchange some of the<br />
potential indigenous parasitoid species for the benefit of both sides. Five parasitoid species;<br />
Aganaspis daci, Diachasmimorpha kraussi and D. tryoni, D. longicaudata and Fopius arisanus<br />
were introduced from Hawaii for evaluation. The parasitoid species have been maintained<br />
under the quarantine conditions for laboratory studies and pilot field trials in Egypt.<br />
Preliminary results showed some success of 2-3 parasitoid species.<br />
Conclusions: Establishment of some of those parasitoid species under Egyptian agroecosystems<br />
will be an additional mortality factor towards the pest control.<br />
Keywords: Bacterocera zonata, Status, Natural enemies, Egypt<br />
115
Session 8<br />
Chemical Ecology, Attractants &<br />
Other Control Methods
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
LETHAL MALE-FEMALE TRANSFER OF INSECTICIDE FROM A FIPRONIL-BASED<br />
MALE ANNIHILATION DEVICE BY BACTROCERA TRYONI (FROGGATT).<br />
Bull, Richard M*<br />
9 Goldieslie Park, Indooroopilly Q4068, Australia. Email: brichard42@optusnet.com.au<br />
Background: Male annihilation technique (MAT) devices are an essential component of<br />
eradication and crop protection programs for Tephritid fruit flies responding to various<br />
parapheromones. In eastern Australia, a mixture of Cue-lure (4-(p-acetoxyphenyl)-2-butanone)<br />
and malathion toxicant is commonly used in MAT devices to attract only the male Queensland<br />
fruit fly (B. tryoni), and on contact, cause rapid death. This paper reports the results of 2 large<br />
field cage experiments that demonstrated male B. tryoni transferred lethal doses of slow acting<br />
fipronil from a new commercial MAT product (Amulet Cue-Lure�) to virgin females during<br />
courtship and mating activities.<br />
Methods: In 2 experiments, 50 sexually mature, virgin, laboratory bred male and female B.<br />
tryoni were released into 3 large (4x4x3 m) field cages, each enclosing a fruiting custard apple<br />
(Anona reticulata :ANONACEA) tree in a commercial orchard. Release was 1.5 hours prior to<br />
their mating time at nightfall and immediately following placement of a single Amulet Cuelure<br />
station in the trees within 2 cages and a non-toxic Cue-lure (control) wick in the 3 rd .<br />
Mortality of flies was determined in both experiments by collection of cadavers from cages for<br />
4 days post-treatment. In the 2 nd experiment an observer in each cage recorded fly behaviour<br />
from time of release until dark. At d-1, 14 female cadavers were collected from the Amulet<br />
MAT cages and analysed by LC/MS/MS for integument surface residues of fipronil.<br />
Results: Both sexes dispersed into foliage on release. After 2 minutes males aggregated on and<br />
around the Cue-lure sources, responding similarly to Amulet MAT and control wicks. Some<br />
spent long periods imbibing Cue-lure and regurgitating repeatedly, but on Amulet MATs<br />
debilitation of engorged males occurred within 15-30 minutes. Others alighted briefly (1-2<br />
minutes) before flying to adjacent leaves, forming groups of 3-10 individuals, leaf-licking and<br />
preening before engaging in aggressive behaviour (head-butting and jostling) to defend a<br />
position for stridulation (pheromone release). At dusk, activity increased, females flying to<br />
higher leaves and males forming a milling swarm above the tree. Females then moved to leaves<br />
near Cue-lure sources where males had stridulated and regurgitated, located a male and<br />
copulated. Mating was observed in all cages, but most in the control. Several females alighted<br />
on an Amulet MAT. In Amulet cages (Exp.1) male and female mortality at 4-d was 97 & 65%<br />
respectively and Cue-lure 28 & 16%. All 14 cadavers analysed (Exp.1 & 2) had fipronil (as<br />
total fiprole) residues of 8-1849pg/mg of fly.<br />
Conclusions: The experiments confirmed slow insecticidal activity of fipronil in the Amulet<br />
Cue-lure device enabled 2 methods of transmission of lethal residues to female B. tryoni to<br />
occur: 1. By direct transfer from contaminated male tarsi during copulation, confirmed by<br />
analyses; 2. By attraction of females to fipronil contaminated locations on leaves (and Amulet<br />
MAT) where males had stridulated and released pheromone.<br />
Keywords: fipronil, Queensland fruit fly, Amulet Cue-lure, MAT.<br />
119
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFICACY TRIALS ON EPA K-LIP®-MF, A NEW ATTRACT&KILL DEVICE FOR<br />
THE CONTROL OF CERATITIS CAPITATA (WIEDEMANN).<br />
de Alfonso, Ignacio* 1 ; Navarro, Ismael 2<br />
1 Ecología y Protección Agrícola, S.L. c/ Gregal, 11; Pol. Ind. Aptdo. de Correos nº 50, Carlet 46240 – Valencia,<br />
Spain; Email: igdeal@epa-ecologia.com; 2 Centro de Ecología y Química Agrícola (CEQA), Instituto Agroforestal<br />
del Mediterráneo (IAM) - Universidad Politécnica de Valencia (<strong>UPV</strong>), Valencia, Spain.<br />
Background: Attract&Kill based control of Medfly has shown to provide excellent results in<br />
fruit crop protection. We report here outlined results of efficacy trials on a new device named<br />
EPA K-LIP®-mF, which possess noteworthy advantages over existing competitors.<br />
Methods: Devices were provided by manufacturer (Ecología y Protección Agrícola, S.L.).<br />
Assays were performed in citrus, stone fruit and persimmon groves in Valencia and Huelva<br />
(Spain). Dose effect was studied for each crop. Plot size for each thesis had a surface of<br />
approximately one hectare. Damage evaluation was carried on over fruit samples that were<br />
kept at evolutionary for 7-10 days before analysis. Female population dynamics was<br />
monitorized employing McPhail traps baited with Biolure®.<br />
Results: Suitable dose for citrus is found to be 50 units/Ha. For stone fruits and persimmon the<br />
proper dose is standing between 100-150 units/Ha depending on sensitivity of fruit variety.<br />
Average fruit damage is below 1% when stated dose are employed. Border effect is clearly<br />
observed. These positive results are remarkable due to high levels of infestation in the<br />
surroundings attending to damage values of blank plots. Spoiled fruit values for standard<br />
chemical treatment plots are clearly higher than those found in EPA K-LIP®-mF treated plots<br />
at recommended doses.<br />
Conclusions: EPA K-LIP®-mF has demonstrated to be a helpful tool for protecting fruit crops<br />
from Ceratitis capitata (Wiedemann) attacks, affording an average damage of less than 1% in<br />
small plots with high pest pressure. This method shown to be more efficient than conventional<br />
chemical treatments and yields residue-free fruits.<br />
Keywords: Medfly, control, field trials, attract and kill, long-life device.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
MONITORING POPULATION DYNAMICS OF ORIENTAL FRUIT FLY WITH WSN<br />
TECHNOLOGY.<br />
Jiang, Joe-Air 1 ; Chen, Chia-Pang 1 ; Lin, Tzu-Shiang 1 ; Liu, Kevin J. 2 ; Ho, Kun-Yaw 3 ;<br />
Chen, Chien-Chung 4 ; Hsu, Ju-Chun 2 ; Yang, En-Cheng 2*<br />
1 Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, Taipei, Taiwan.<br />
2 Department of Entomology, National Taiwan University, Taipei, Taiwan. Email: ecyang@ntu.edu.tw; 3 Chia-Yi<br />
Agricultural Experiment Station, Agricultural Research Institute, Chia-Yi, Taiwan. 4 Agricultural Research<br />
Institute, Council of Agriculture, Taichung, Taiwan<br />
Background: Traditional monitoring method on the oriental fruit fly (OFF) population is based<br />
on poisoned methyl eugenol trap and labor-intensity counting work. This study adopts wireless<br />
sensor network (WSN) technology which combines sensor modules and wireless signal<br />
transmission to develop a specialized sensor module to detect the attracted OFFs. With the<br />
monitoring system, a sub-hour timescale monitoring database is accomplished, and the wireless<br />
signal transmission network allows the distribution of a hectare-wide sensor array to monitor<br />
the spatial distribution of the pests.<br />
Methods: The WSN technology was applied to build up an ecological monitoring system for<br />
monitoring the OFF population in the Chiayi county, Taiwan since April, 2009. Twenty<br />
poisoned methyl engenol traps were deployed as a 5 � 4 array in an orchard (25 m × 50 m).<br />
Each of the traps was coupled with a two-paired infra-red module specially designed for<br />
counting the male fly number automatically and equipped a set of sensor node to record the<br />
environmental factors as well. The recorded data was then transmitted to a local gateway by<br />
wireless network and to the database by Global System of Mobile Communication (GSM)<br />
module as a short message (SM) every 30 minutes. The auto-counting data of the traps were<br />
individually counted by human twice a week for checking the accuracy of the system.<br />
Results: The year-round population variation just like most surveys, the population climbs high<br />
by May, drops in October and keeps low to the following April. However, the hourly<br />
population data shows that the male fruit fly is active in the daytime. The earliest trapped<br />
individuals are positive correlated to sunrise time, and more individuals are attracted in the<br />
morning than afternoon. In terms of spatial distribution, more than 40% of the trapped flies<br />
were constantly captured by the most western 4 traps of the orchard through the year. Our<br />
results suggest that the OFFs reside in and come directionally from the nearby.<br />
Conclusions: Using the specialized fruit fly sensor module and WSN technology, a nearly realtime<br />
monitoring system for the methyl eugenol-attracted male OFF has been accomplished.<br />
The hourly data reveals that the fruit flies are attracted in the daytime and mostly in the<br />
morning. This auto-counting system provides easier and detailed population data than ever<br />
before. Furthermore, grouped sensor array represents the coming direction of local population.<br />
Some hot-spots appear in the survey area and the trapped flies of the most western 4 nodes<br />
represent more than 40% of the total surveyed number. With the powerful spatial and temporal<br />
resolution of population survey provided by the WSN technique, getting insight of the pest<br />
population dynamics for future pest control is promising.<br />
Keywords: Bactrocera dorsalis, wireless sensor network, population dynamics, methyl eugenol.<br />
121
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EXPLORING STRUCTURE-ACTIVITY RELATIONSHIPS IN THE PHENYLPROPANOIDS<br />
TO PROCURE NEW MALE LURES FOR NON-RESPONSIVE BACTROCERA AND DACUS.<br />
Fay, Harry A. C.<br />
Horticulture and Forestry Science, Agri-Science Queensland, Dep. of Employment, Economic Development and<br />
Innovation, 28 Peters Street, Mareeba Q. 4880 Australia. Email: harry.fay@deedi.qld.gov.au<br />
Background: It is generally thought that between 30 and 50% of Bactrocera and Dacus spp. do<br />
not respond to either of the two recognised male lures, cuelure (CL) and methyl eugenol (ME).<br />
Included among these species are pests of quarantine or market access importance, such as<br />
Bactrocera atrisetosa, B. cucumis, B. decipiens and B. jarvisi. Regulatory authorities are under<br />
increasing pressure to report on such pests, making an ability to monitor activity and to use<br />
male annihilation as a control/eradication option clearly desirable. While several past studies<br />
have looked at structural variants of CL and ME for B. cucurbitae and B. dorsalis respectively,<br />
there has been little research on new male lures for non-responding species in Dacinae endemic<br />
areas.<br />
Methods: The study reported here examined 50 different phenylpropanoids (selected on their<br />
chemical structures) in a series of trials at a field site in north-east (tropical) Australia, and<br />
mostly compared their attractiveness to Bactrocera and Dacus spp. against raspberry ketone<br />
(RK) (the hydrolysed form of CL) and ME. The phenylpropanoids were applied to dental<br />
wicks with maldison in 1L modified Steiner traps and left in the field for 6-9 weeks during the<br />
wet season.<br />
Results: Ten of the 50 compounds tested did show activity, which varied from weak to very<br />
strong attraction. Zingerone (4-hydroxy, 3-methoxyphenyl-2-butanone), which structurally lies<br />
between RK and ME, was highly attractive to B. jarvisi. It also attracted small numbers of<br />
some CL species, the non-lure responding B. aglaiae, the rarely caught Dacus secamoneae and<br />
a totally new species, B. speewahensis. The non-lure responding B. aurea also came to this<br />
compound in a related study in south-east Queensland. Methyl-3-(4-hydroxyphenyl) propionate<br />
was the next most powerful lure and attracted a similar range of species (11) to RK (14). If<br />
differs structurally from RK in having an additional oxygen molecule in the primary<br />
substituent attached to the phenyl ring i.e. methyl propionate instead of 2-butanone. This<br />
compound was substantially more attractive than anisylacetone, a precursor of CL. Twomethoxy-4-propyl<br />
phenol was the only compound that attracted both B. jarvisi and B. visenda<br />
(a ME species), and the only one that didn’t possess a double bond in the primary substituent.<br />
Both isoeugenol and methyl isoeugenol are structurally similar to ME but were less attractive<br />
to those species that came to the latter. However, isoeugenol attracted a species that is possibly<br />
new although similar to B. kraussi (a CL species), while methyl isoeugenol attracted B.<br />
nigrovittata, a non-lure responding species only known previously from New Guinea.<br />
Conclusions: A number of phenylpropanoids have been identified which are attractive to a<br />
range of Bactrocera that do not respond to either CL or ME and further exploration of their<br />
potential through the Asia-Pacific region could prove profitable.<br />
Keywords: Dacinae, phenylpropanoids, new male lures, Australia
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IDENTIFICATION OF ROOSTING HOSTS AND THEIR INTEGRATION INTO BAIT<br />
APPLICATIONS FOR THE MANAGEMENT OF CUCURBIT FLIES IN WEST AFRICA.<br />
Hanna, Rachid 1 *; Azandémé, Ginette 1 & Gnanvossou, Desiré 1<br />
1 International Institute of Tropical Agriculture (IITA), Cotonou, Benin; 08 BP 0932 Tri Postal, Cotonou Benin.<br />
Email: r.hanna@cgiar.org.<br />
Background: Several types of protein baits have been used successfully in fruit fly<br />
management. In some cases, roosting hosts have been integrated into bait spray programs. Here<br />
we report on experiments that identified the most appropriate roosting hosts for cucurbitinfesting<br />
fruit flies in Benin (West Africa) and their use as border plants for the spot<br />
applications of GF-120.<br />
Methods: Two experiments were conducted at IITA station near Cotonou, Benin. In the first<br />
experiment, three main plots (30m x 30m), separated by at least 100m were used. The main<br />
plots were planted with zucchini and watermelon. Twelve potential roosting hosts were planted<br />
in 5m x 5m plots along the borders - three roosting host plots equally spaced on each side of<br />
main plots. Abundance and identity of adult fruit flies were recorded during 10-min visits twice<br />
a day (starting at 1200h and at 1800h) at weekly intervals over a period of 2.5 months. Fruit fly<br />
infestations of cucurbit fruits were evaluated at two-week intervals. Sorghum and castor bean<br />
were selected as most suitable roosting hosts for integration with GF-120 spot sprays in an<br />
experiment that followed a design similar to the first experiment, but with six plots – three for<br />
GF-120 and three for control. The two roosting hosts were planted in alternating fashion in the<br />
four corners of each of the cucurbit plots. Plots were separated by at least 100m. Spot<br />
applications of GF-120 at the rate of 2.5 l/ha of cucurbits (1 litre GF-120:1.5 litre water) were<br />
carried out at weekly intervals (0700h-0800h) starting at 25 days after planting of cucurbits.<br />
Fruit fly monitoring schedules on cucurbits and resting host was similar to the first experiment.<br />
Results: Of the 12 roosting hosts, castor bean was consistently the most suitable for all five<br />
species of cucurbit fruit flies encountered in the study site, followed by sorghum; but the latter<br />
was most attractive when it was infested with aphids. GF-120 spot sprays were much more<br />
attractive to fruit flies on castor bean compared with sorghum. Fruit fly infestations on zucchini<br />
and watermelon in GF-120 plots were reduced to 77% and 69% respectively. Dacus ciliatus<br />
and D. vertebratus were the dominant fruit flies infesting zucchini and watermelon<br />
respectively. Other fruit flies included B. cucurbitae, D. punctatifrons and D. bivittatus.<br />
Conclusions: GF-120 spot applications on castor bean border plants can be successfully used in<br />
integrated management of tephritid fruit flies in cucurbit crops in Benin and likely other<br />
countries in West Africa. Large economic returns can be expected from the recommended<br />
system because of the high losses in unprotected cucurbits and the efficacy of GF-120 in<br />
reducing fruit fly infestations.<br />
Keywords: GF-120, protein bait, Dacus spp., Bactrocera cucurbitae, integrated management<br />
123
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE INFLUENCE OF THE PHYSIOLOGICAL AGE OF CERATITIS CAPITATA<br />
(WIEDEMANN) FEMALES ON THEIR PREFERENCES FOR TWO MASS TRAPPING<br />
SYSTEMS.<br />
Restrepo-Ortiz, Claudia X. *1 ; Botta, Anna 2 ; Marín, Cándido 2 ; Pujade-Villar, Juli 1<br />
1 Universidad de Barcelona (UB), Facultat de Biologia, Departament de Biologia Animal, Av. Diagonal 645,<br />
08028 Barcelona (Spain). Email: claudiaximenaro@gmail.com. 2 R&D Plant Physiology Division, BIOIBERICA,<br />
S.A, Palafolls, Barcelona (Spain).<br />
Background: Mass trapping techniques have been consolidated as an effective solution to<br />
control Medfly Ceratitis capitata (Diptera: Tephritidae) worldwide. One of the reasons for<br />
their success is the strong attraction that ammonia odor exerts to the females, which are the<br />
damaging fruit agent and the dominant factor for pest multiplication. Among females, it is<br />
believed that the youngest are more attracted to proteinic compounds due to the need for an<br />
external supply of protein for ovary maturation. With these findings as background, this report<br />
further evaluates the influence of the physiological age of the Medfly female on the attraction<br />
to two different kinds of mass trapping systems.<br />
Methods: We performed three different crop trials (fig, pome granate and mandarin trees)<br />
during Medfly season in the south-east of Spain in order to compare the effectiveness of two<br />
mass trapping systems; a solid attractant which contains ammonium acetate, trimethylamine,<br />
putrescine and that requires DDVP insecticide (named Standard) and a liquid protein<br />
formulation obtained by enzymatic hydrolysis and free of pesticides (Cera Trap � ). In each trial,<br />
system effectiveness was evaluated during three weeks by assessing: (i) the number of<br />
captures, (ii) percentage of females and males and (iii) percentage of different physiological<br />
females in each system. In order to classify the physiological age of females, we studied their<br />
internal reproductive organs through dissection and three categories of ovarium development<br />
were established: pre-ovoposition stage, ovoposition stage and senile ovarium.<br />
Results: Total Medfly captures per trap in the three trials performed were significantly higher<br />
with the Cera Trap system than with the Standard one. In both systems, the percentage of<br />
females was three times larger than males; however, regarding the higher number of total<br />
captures of Cera Trap, more females were captured with this system in the three trials.<br />
Concerning the physiological age classification, pre-ovoposition and mature flies were highly<br />
captured in both Cera Trap and Standard, which implies a significant reduction of the pest<br />
pressure for the next generations. The capture of senile flies was considered accidental in both<br />
systems. The percentage of total pre-ovopositing females shows a statistical significantly larger<br />
amount of captures of these younger flies with the Cera Trap system (60%) compared with the<br />
Standard system (40%).<br />
Conclusions: Both mass trapping systems show a great performance on the control of Medfly<br />
populations due to the large number of captures per trap during the whole Medfly season,<br />
especially for the higher percentage of female flies captured. However, this study shows that<br />
Cera Trap system is the most effective, not only for capturing a greater number of Medflies,<br />
but especially for its attraction effect on young flies, thus providing control of Medfly<br />
population before females are able to damage fruits and multiply the pest population.<br />
Keywords: Medfly, Mass trapping, Ceratitis capitata, physiological age
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
USE OF LIVE-CAUGHT INSECTS EXPANDS THE SCOPE OF TRAPPING.<br />
Kouloussis, Nikos* 1 ; Papadopoulos, Nikos 2 ; Katsoyannos, Byron 1 & Carey, James 3<br />
1 Aristotle University of Thessaloniki, School of Agriculture, Department of Plant Protection, Laboratory of<br />
Applied Zoology and Parasitology, 54124 Thessaloniki, Greece; Email: nikoul@agro.auth.gr; 2 University of<br />
Thessaly, Depart. of Agriculture Crop Production and Rural Environment, Laboratory of Entomology and<br />
Agricultural Zoology, Volos, Greece; 3 University of California, Depart. of Entomology, Davis, CA, USA.<br />
Background: Insect traps are commonly used to draw a sample from wild populations and then<br />
use information gained from this sample to infer parameters that apply to the entire population.<br />
Insects are almost always killed during the process of trapping, so the datum used is the<br />
number of dead flies captured. Extrapolations are then made regarding the actual size of the<br />
population and a number of other parameters. Several types of traps, however, can be modified<br />
to capture live insects from which we may then derive additional valuable “latent” information<br />
on biology and behavior.<br />
Methods: In recent studies [e.g. J. R. Carey et al., Aging Cell 7: 426–437 (2008)], Ceratitis<br />
capitata (Diptera: Tephritidae) were trapped alive with the use of McPhail-type traps baited<br />
with a food-based synthetic attractant. The survival patterns of wild-caught individual insects<br />
of unknown age were then used to estimate age structure in the populations from which these<br />
individuals were derived. Several important findings were obtained from that study. For<br />
example, it was found that major changes in age structure occur in wild populations of C.<br />
capitata and that a substantial fraction of individuals survive to middle age and beyond. The<br />
time of capture during the season had a significant effect on the captive lifespan, the average<br />
daily reproduction and on the patterns of egg-laying.<br />
Conclusions: The above example provides an illustration of how live-caught insects may be<br />
used in trapping as a means of extracting information that cannot be obtained by dead insects.<br />
Live-caught insects have a history (related to food, symbiont acquisition, mating, exposure to<br />
hardiness, etc) and are loaded with information related not only to aging and reproduction but<br />
also to several other parameters in their biology and behavior. Certain of this latent information<br />
could be derived in the lab post-capture, thus considerably advancing the scope of sampling<br />
science.<br />
Keywords: Trapping, sampling, reproduction, age structure, Ceratitis capitata, natural population<br />
125
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFICACY ASSESSMENT OF BAIT STATIONS AS A CONTROL METHOD FOR<br />
CERATITIS CAPITATA (WIEDEMANN) IN FIELD TRIALS.<br />
Navarro-Llopis, Vicente*; Sanchis, Juan; Vacas, Sandra, Alfaro Cañamás, Cristina;<br />
Primo, Jaime<br />
Centro de Ecología Química Agrícola - Instituto Agroforestal del Mediterráneo. Universidad Politécnica de<br />
Valencia. Camino de Vera s/n. 46022. Valencia. Spain. Email:<br />
Background: Mass trapping or lure and kill methods have been widely used to control fruit fly<br />
populations during last century. But the efficacy of these techniques depends on the efficacy of<br />
female attractants and the cost and easy to hand properties of the devices. Using high efficacy<br />
female attractants the number of required devices can be reduced and, therefore, the<br />
replacements of devices that increase the final cost can be prevented. Moreover a well designed<br />
device reduces hand labor and increase the killing efficacy. In this work we report the efficacy<br />
of new designed bait station devices with more long lasting and efficient female attractants in<br />
field trials.<br />
Methods: Two types of bait stations were tested: a chemosterilant one with lufenuron in the<br />
bait using 24 traps per ha, and an insecticide bait with cypermethrin applying between 50 and<br />
100 devices per ha. Field trials were conducted in citrus, persimmon and peach orchards and<br />
for efficacy assessment fruit fly population and fruit damage before and during harvest were<br />
evaluated. Chemosterilant treatment was compared with malathion aerial bait sprays and<br />
insecticide baits were compared with terrestrial sprayed insecticides and mass trapping<br />
technique.<br />
Results: Chemosterilant devices reduced significantly fruit fly populations regard malathion<br />
aerial treatment since the first year of application, but the best results were obtained in the third<br />
and fourth year after chemosterilant treatment, when a reduction of 80% of C. capitata<br />
population was reached. Fruit damage showed a significant reduction in persimmon in the third<br />
and fourth year. C. capitata populations were also reduced significantly in field with<br />
insecticide baits regard insecticide sprayed fields. Fruit damage was maintained below 1% in<br />
bait stations treated fields, without significant differences with mass trapping fields. However,<br />
in untreated fields, fruit damage was over 25%.<br />
Conclusions: Bait stations were efficient enough to represent an alternative to chemical control<br />
with insecticides. In addition, the effect of bait station is long lasting and therefore cumulative<br />
effect could be observed if wide areas are treated with this technique. Finally, bait stations add<br />
several advantages like non residues in fruit, specificity affectation of target pest and avoid<br />
insecticide exposure by applicators.<br />
Keywords: Ceratitis capitata, chemosterilization, bait station, lure and kill.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SPLAT-MAT SPINOSAD ME A NEW CONTROL STRATEGY FOR INVASIVE<br />
BACTROCERA SPECIES.<br />
Gomez, Luis E.* 1 ; Mavrotas, Costas 2 ; Lysandrou, Michael 2 ; Paroonagian, Doris 1 ;<br />
Vargas, Roger, 3 Borges, Rafael 4 & Mafra-Neto, Agenor 5<br />
1 Dow AgroSciences LLC 9330 Zionsville Road, Indianapolis, IN 46268; Email: egomez2@dow.com; 2 Dow<br />
AgroSciences Lavrion, Greece. 3 USDA, ARS, HI USA. 4 ISCA Technologias Ltda, Brazil. 5 ISCA Technologies,<br />
INC. CA USA.<br />
Background: Tephritid fruit flies include some of the world’s most destructive insect pests.<br />
Areas infested with fruit flies are quarantined and fruit cannot be shipped to non-infested areas<br />
without expensive radiation, cold, or heat treatments that can reduce fruit quality and shelf life.<br />
Fruit fly control tactics include the use of cover sprays, toxicant-laced baits, and Male<br />
Annihilation Techniques (MAT) that still rely on pyrethroid and organophosphate (OP)<br />
insecticides. Public concern over the use of these actives has grown around the world.<br />
Methods: Dow AgroSciences and ISCA Technologies embarked in developing a sprayable<br />
novel MAT that would match performance to existing pyrethroid and OP-based options.<br />
Spinosad, a green chemistry insecticide, is the toxicant and methyl eugenol (ME) is the<br />
attractant. SPLAT, a monolithic passive dispenser, is the basis of the formulation that protects<br />
the components from the environment (e.g. rain and UV light) and provides a metered release<br />
of the attractant and toxicant, making them available for longer time. This novel product<br />
attracts male fruit flies from long distances making it suitable for off-crop applications thus<br />
eliminating pesticide and attractant residues on the fruit.<br />
Results: Product was evaluated in Hawaii, USA to assess residual, and rainfastness. SPLAT-<br />
MAT Spinosad ME and Min-U-Gel-naled MAT were aged for up to 12 weeks with average<br />
monthly rainfall of 158 mm during the trial period. Samples were exposed weekly to endemic<br />
oriental fruit fly (Bactrocera dorsalis) populations. The novel MAT had similar performance to<br />
Min-U-Gel-naled MAT during the first 6 weeks and outperformed it from weeks 7-12. In<br />
Brazil, two large trials were established to compare one single application of this product to the<br />
local MAT standard, wooden blocks dipped in a solution of malathion+ME applied every 20<br />
days, against the carambola fruit fly (Bactrocera carambolae). A single application of SPLAT-<br />
MAT Spinosad ME outperformed multiple applications of the standard treatment during 3<br />
months. In Pakistan a single application of this technology outperformed weekly applications<br />
of the standard cover spray, Dipterex, for the peach fruit fly (Bactrocera zonata) control in two<br />
trials.<br />
Conclusions: SPLAT-MAT Spinosad ME, a novel green chemistry-based MAT was<br />
developed and is currently registered in Brazil and the USA. It has been tested against three<br />
Bactrocera species in different geographies. Within each scenario the product outperformed<br />
the local standard MAT or cover spray program. Fewer applications were required to regulate<br />
male and subsequent female populations while resulting in reduced damaged and infested fruit.<br />
Benefits from this technology are: reduction in application costs, elimination of insecticide<br />
residues on fruit, increased marketable fruit, and reduction of infested fruit being transported to<br />
fruit fly free zones.<br />
Keywords: MAT, Spinosad, oriental fruit fly, peach fruit fly, carambola fruit fly.<br />
127
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ATTRACTION AND FEEDING RESPONSES OF THREE CERATITIS SPECIES TO<br />
PROTEIN BAITS.<br />
Manrakhan, Aruna*; Kotze, Charl & Daneel, John-Henry<br />
Citrus Research International, PO Box 28, Nelspruit, 1200, South Africa; E-mail address: aruna@cri.co.za<br />
Background: Recommendations for fruit fly control using baits are usually generic. Insecticidal<br />
baits applied as sprays or stations are often assumed to result in the same level of control<br />
irrespective of pest fruit fly species. In South Africa, three congeneric species: Ceratitis<br />
capitata, C. rosa and C. cosyra co-exist in the north/north east and eastern areas. The attraction<br />
and feeding responses of the three species to proteinaceous baits used in fruit fly control were<br />
evaluated in field cages and laboratory bioassays respectively.<br />
Methods: Mature protein-deprived, laboratory-reared C. capitata, C. rosa and C. cosyra were<br />
used in all tests. Species were evaluated separately. Attractiveness of HymLure, GF 120<br />
Naturalyte and M3 bait (also known as Questlure and used in the M3 bait station registered for<br />
fruit fly control in South Africa) was evaluated in field cages. A novel method of bait<br />
evaluation was used where baits tested were each placed in an aerated plastic container which<br />
was in turn suspended in an open-ended plastic cylinder lined with a sticky insert to trap<br />
attracted flies. Feeding responses of HymLure, Solbait and M3 bait were evaluated separately<br />
in the laboratory in aerated plastic containers fitted with a removable glass top. Female flies<br />
were released in the containers one hour before bait exposure. All baits were combined with a<br />
green food dye at the rate of 1%. Baits were exposed as dried droplets (a drying period of 30<br />
minutes at 30° C) on glass slides which were weighed before exposure to flies. Feeding tests<br />
ended after 24 hours when the number of flies containing the green dye was counted and the<br />
glass slide containing the bait was weighed to determine the amount of bait consumed per<br />
female.<br />
Results: Attraction responses to baits were species-specific. C. cosyra had a low response to<br />
HymLure but responded well to the other baits. C. capitata was the most responsive of all<br />
species tested. There was variability in consumption between baits for each species and<br />
between species for similar baits.<br />
Conclusions: The study showed differences in responses to protein baits even between closely<br />
related congeneric species. As such, in areas of multiple fruit fly pest species, it is essential to<br />
choose the appropriate bait or combinations of bait that will work effectively against all species<br />
present.<br />
Keywords: C. capitata, C. rosa, C. cosyra, baits
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
USE OF MAGNET MED TECHNOLOGY FOR THE ATTRACT AND KILL OF<br />
MEDITERRANEAN FRUIT FLY (CERATITIS CAPITATA).<br />
Casagrande, Enzo*. & Martí, Salvador<br />
Suterra Europe Biocontrol, S.L. Director of Technology. C/ Argenters, 5. Parc Tecnològic del Vallès 08290<br />
Cerdanyola del Vallès. Spain. enzo.casagrande@suterra.es<br />
Background: The optimization of potent Medfly attractants identified by the USDA has<br />
revolutionized the strategies used for the control of this important pest of fruit crops replacing<br />
both chemical cover sprays and bait sprays. Till now, however, these attractants have been<br />
formulated for use mainly for the mass trapping of Medfly where they applied in combination<br />
with suitable traps and killing agents. While very effective these systems are cumbersome and<br />
costly. Magnet MED moves this technology on to the next level by combining the attractants;<br />
the target device and the killing agent into a single ready to use attract and kill (A&K) system.<br />
Methods: Developed and tested over the last 4 years, Magnet MED offers a highly effective<br />
system for the control of Medfly. The presentation will detail the development of the<br />
technology and will present the results of trials carried out mainly in Italy and Spain. These<br />
demonstrate that A&K gives control equal or better than the current mass trapping systems and<br />
is as effective as any conventional insecticide program used for Medfly control.<br />
Results: The system uses very low rates of Deltamethrin insecticide at between 0.5 to 0.75g per<br />
hectare for season long control up to 5 months. The system has been optimized so that the<br />
application of 50 devices per hectare can now be recommended for most crops with the option<br />
to increase to 75 per hectare in vulnerable crops such as peaches. Because the insecticide is not<br />
applied directly to the crop and can be removed at the end of the season, the formulation is<br />
approved for use in organic orchards in the European Union.<br />
129
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SPINTOR CEBO ®<br />
(GF-120 ® ), A UNIQUE CONTROL STRATEGY FOR<br />
SUSTAINABLE FRUIT FLY MANAGEMENT IN SPAIN.<br />
Torné, Maria 1* & Mavrotas, Costas 2<br />
1 DowAgroSciences Ibérica, Ribera del Loira 4-6 4ª, 28042 Madrid (Spain); Email: mtorne@dow.com;<br />
2 DowAgroSciences, Lavrion, Greece.<br />
Background: SPINTOR ® CEBO (GF-120 ® , SUCCESS ® , SPINTOR ® FLY), is a unique premixed<br />
fruit fly bait containing spinosad, an organic insecticide, and an optimized blend of<br />
feeding attractants. It offers improved efficacy and attractiveness with a favorable<br />
environmental and toxicological profile when compared to current toxicant bait mixtures.<br />
SPINTOR ® CEBO attracts and controls males and females of multiple species of tephritid fruit<br />
flies. SPINTOR ® CEBO delivers very low quantities of insecticide per ha (0.24-0.36 grams)<br />
decreasing environmental load by 600 to 8,000 times per season when compared to<br />
conventional fruit fly control products commonly used in Spain. Spinosad is derived from the<br />
fermentation of a soil bacterium, Saccharopolyspora spinosa, with a unique mode of action on<br />
the insect nervous system. SPINTOR ® CEBO is currently the only pre-mixed bait authorized in<br />
Spain for ground and aerial applications in citrus and olives.<br />
Methods: Field, laboratory research, and commercial applications over the past 10 years in<br />
Spain in citrus and olives are summarized. Applications were done using ground or air,<br />
equipment customized to achieve optimum spray parameters.<br />
Results: Our experience with SPINTOR ® CEBO clearly demonstrates that in order to obtain<br />
the maximum attraction and residuality the product must be applied as a concentrated bait at<br />
rates of 1 – 1.5 L product/ha in 4 to 10 liters of total mixture (with water). The application<br />
should deliver droplet sizes of 4-6 mm and should be aimed to the top part of the trees, where<br />
adult flies are more active. Ground application equipment was developed to deliver SPINTOR ®<br />
CEBO at the right rates and recommendations resulting in a significant reduction of spray<br />
volume (90-95%) versus conventional bait sprays. Since this application method represented a<br />
significant change for farmer practices, an extensive training program was implemented. Aerial<br />
applications have been adapted resulting in a reduction of spray volume of more than 50%,<br />
reducing the total cost and time to treat crops.<br />
Conclusions: Extensive experience on efficacy against Ceratitis capitata in citrus and<br />
Bactrocera oleae in olive has been gathered in the last decade in Spain. When used according<br />
to application recommendations, SPINTOR ® CEBO consistently delivers excellent efficacy for<br />
fruit fly control and crop selectivity. SPINTOR ® CEBO provides key attributes to the growers<br />
such as lower application cost, short pre-harvest interval (1 day), EU and USA authorization in<br />
Organic farming, favorable environmental and toxicological profile, and compatibility with<br />
beneficial arthropods and pollinators. SPINTOR ® CEBO should be a key component of an<br />
integrated fruit fly control strategy for a sustainable agriculture to meet present and future<br />
farmer and export market needs.<br />
Keywords: Medfly, olive fruit fly, spinosad bait, organic, aerial application, efficacy
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
TRITROPHIC INTERACTIONS IN THE COMPLEXES OF FRUIT FLIES DAMAGING<br />
FRUIT AND VEGETABLE CROPS IN LA REUNION ISLAND<br />
Quilici, Serge*; Duyck, Pierre-François; Brévault, Thierry; Rousse, Pascal; Jacquard, Cathy; Delatte,<br />
Hélène; Deguine, Jean-Philippe; Lereculeur, Anabel; Wattier, Christopher; Atiama, Toulassi; Hurtrel,<br />
Béatrice; Franck, Antoine; Simiand, Christophe & Chiroleu, Frédéric<br />
Centre de coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), Département<br />
des Systèmes Biologiques, Saint-Pierre (La Réunion), France; E-mail: serge.quilici@cirad.fr<br />
Background: The study of tri-trophic interactions is a vast field of basic ecological studies<br />
which are also of paramount importance for the development of environment-friendly methods<br />
of pest management. Within the Diptera, the Tephritidae family represents an interesting model<br />
for such studies because of the variable host-specificity of species in this group, and the<br />
important role that some natural enemies may play in the regulation of their populations.<br />
Furthermore, the great economic importance of many species in this family offers a large array<br />
of applications to research results. La Réunion, a French island situated in the south-west of the<br />
Indian Ocean, is a favourable area for studying such interactions, because of the number of<br />
tephritid species present in the island, and the great variability of its climatic conditions. On<br />
fruit crops, a complex of three polyphagous species of tephritids cause considerable damage:<br />
the Mediterranean fruit fly, Ceratitis capitata, the Natal fruit fly, C. rosa, and the Peach fruit<br />
fly, Bactrocera zonata. In addition, another complex of Dacini is harmful to cucurbit crops: the<br />
Melon fly, B. cucurbitae, the Ethiopian cucurbit fly, Dacus ciliatus, and the Indian Ocean<br />
cucurbit fly, D. demmerezi. Conversely, on Solanaceous crops such as tomato, the Tomato fruit<br />
fly Neoceratitis cyanescens, is the only species of economic importance.<br />
Methods: Over the last twenty years, many field studies allowed us to specify the host-range<br />
and relative importance of the different species on cultivated crops. Of particular interest are<br />
some host preferences observed in the field in some otherwise very polyphagous species, such<br />
as C. capitata. Detailed studies (lab, wind tunnel and field cages) were also devoted to the host<br />
location behaviour of stenophagous species, taking as a model the tomato fruit fly, N.<br />
cyanescens. Other behavioural studies also highlighted the preferences of the different species<br />
for particular host-plants during foraging or egg-laying behaviour. More recently, studies were<br />
focused on the quality of various host or non-host fruits for the pre-imaginal development of<br />
the different species, and its influence on their fitness. Though some indigenous parasitoids of<br />
the Mediterranean fruit fly have been recorded in the island, most of the natural regulation by<br />
parasitoids is due to exotic species imported through classical biological control programmes.<br />
This is particularly the case with two species imported from Hawaii, in collaboration with<br />
USDA Hawaii and the University of Hawaii: Psyttalia fletcheri, a larvo-pupal parasitoid of the<br />
Melon fly, and, more recently, Fopius arisanus, an egg-pupal parasitoid of some Bactrocera<br />
spp. Following the acclimatization of these two species, field studies allowed us to evaluate<br />
their host range, favourite habitats, and impact on host species populations. In addition,<br />
laboratory, field-cage and wind tunnel studies improved our knowledge of the stimuli involved<br />
in host habitat and host selection behaviours.<br />
Results: Results of these studies are summarized and discussed in relation to the current state<br />
of knowledge of insect-plant and host-parasitoid interactions in tephritids, and to their possible<br />
applications in pest management.<br />
Keywords: Tephritidae, tri-trophic interactions, insect-plant relationship, host-parasitoid relationship.<br />
131
Workshop<br />
Longevity, Ageing & SIT
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
A HIGH-RESOLUTION SYSTEM FOR RECORDING THE DAILY AND LIFETIME<br />
BEHAVIORAL AND MOVEMENT PATTERNS OF INDIVIDUAL MEXICAN FRUIT FLIES.<br />
Carey, James R. 1 ; Zou, Sige 2 ; Altamirano, Leopoldo 3 ; Ingram, Donald 4 & Liedo, Pablo 5 *<br />
1 Department of Entomology, University of California Davis, Davis CA 95616 USA; 2 Laboratory of Experimental<br />
Gerontology, National Institute on Aging, Baltimore MD 21224 USA; 3 Laboratorio de Visión por Computadora,<br />
Instituto Nacional de Astrofísica, Óptica y Electrónica, Tonanzintla, Pue. México; 4 Pennignton Biomedical<br />
Research Center, Louisiana State University, Baton Rouge LA 70808 USA; 5 Departamento de Entomología. El<br />
Colegio de la Frontera Sur (ECOSUR), Tapachula, Chiapas, México; Email: pliedo@ecosur.mx<br />
Background: We developed a system in which the lifetime behavior and movement patterns of<br />
the Mexican fruit fly, Anastrepha ludens, maintained in 27 cm 3 cages is recorded every fifth of<br />
a second for one minute three times per hour for 24 hours each day throughout their lives. The<br />
data generated from the behavioral monitoring system (BMS) include date, time, 1-of-6<br />
behaviors, and locations in 3-dimensional space (XYZ location). This yields over 12 million<br />
data-points for a single fly that lives 100 days. The raw data enable researchers to analyze: (1)<br />
behavior including frequency, sequence, age and diel patterns; (2) movement including age and<br />
diel patterns, rates, and pathways; and (3) location within the cage by age and time-of-day.<br />
Methods: The hardware of the BMS consists of a pair of cameras, a nine-cage tray to house<br />
individual flies in each cage, visible and infrared lights. The cameras are connected to an image<br />
processor for short term storage of video images, which are fed to a computer for behavior<br />
detection and long-term data storage. The BMS records 3-D coordinates of individual flies and<br />
automates classification of six behaviors, termed as resting, moving, walking, flying, feeding<br />
and drinking. Two BMS were used to monitor lifetime behaviors of 16 A. ludens females with<br />
each BMS holding four females on the full diet and four females on sugar only diet. The<br />
recording was initiated at the eclosion of the flies and stopped at day 134 when approximately<br />
half of the flies were dead.<br />
Results: Flies on both diets spend more than 60% of their time on resting every day over their<br />
lifetime. The next most frequent daily behaviors were walking and moving, which, combined,<br />
counted approximately 25% of a fly’s daily time. Flies on sugar diet had significantly higher<br />
walking frequency than those on the full diet. Flying, feeding and drinking occured at very low<br />
frequencies, which counted less than 5% of all the behaviors in most of the days. The<br />
frequency of daily walking reached the peak around the age of 20 days and then gradually<br />
decreased with age for flies on both diets, suggesting that walking frequency is an age-related<br />
parameter. Overall, the frequency of daily flying declined with age after reaching the peak<br />
around the age of 20 days, while the frequency of daily feeding and drinking appear to<br />
increased with age. During night time between 7 pm when the light is off and 7 am when the<br />
light was on, flies spent almost all of their time (>95%) on resting, presumably in sleep, while<br />
other behaviors were rare. Walking and flying showed hour-dependent changes and reached<br />
the peak period in late afternoon from 3-7 pm. The daily total distance gradually increased in<br />
the first 15 days, reached the peak at the age of approximately 20 days and then gradually<br />
declined with age for flies on both diets.<br />
Conclusion: This BMS can be easily adopted for a regular research lab. It is the first high<br />
resolution machine vision system to have the capacity to record lifetime 3-D positions of an<br />
animal and automatically classify lifelong behaviors for months.<br />
Keywords: Anastrepha ludens, lifetime behavior, aging, biodemography.<br />
135
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DIFFERENTIAL RESPONSE OF A LONG AND A SHORT-LIVED MEDFLY POPULATION<br />
TO LARVAL CROWDING.<br />
Diamantidis, Alexandros 1,2 ; Carey, James R. 2 & Papadopoulos, Nikos T. 1<br />
1 Laboratory of Entomology and Agricultural Zoology, Department of Agriculture, Crop Production and Rural<br />
Environment, University of Thessaly Greece, Phytokoy St. Nea Ionia (Volos), 38446 Magnisia, Greece; Email:<br />
nikopap@uth.gr; 2 Department of Entomology, University of California, Davis CA 95616<br />
Background: Fruit fly larval competition, within infested fruits, may arise because of increased<br />
population densities. This is a form of scramble, intra-specific competition that may affect vital<br />
life history traits for immature and adult stages. Older studies have addressed effects of larval<br />
crowding on aspects of development and survival of immatures. Nevertheless, little is known<br />
regarding the effects of larval crowding on adult life history traits. And, it still remains<br />
unknown if different populations (biotypes) selected for short or long adult lifespan exhibit the<br />
same response under stressful conditions imposed by larval crowding. Here we investigated<br />
the effect of larval crowding on the life-history traits of a long and a short-lived Mediterranean<br />
fruit fly, Ceratitis capitata, (Wiedemann; Diptera: Tephritidae) populations.<br />
Methods: Experiments were conducted during 2006 – 2007 under constant laboratory<br />
conditions (25 ± 1 °C, 65 % R.H., 14:10 L: D). We used a long, and a short-lived medfly<br />
population originating from Greece and Guatemala respectively. Newly hatched larvae of the<br />
two populations were randomly assigned in six treatments in relation to larval density: 1, 5, 15,<br />
30, 60, and 120 larvae per 3 ml of larval diet. We recorded larval and pupal survival, pupal size<br />
(length and weight), and duration of larval and pupal development. Soon after adult emergence<br />
pairs consisting of a male and a female from four (1, 15, 60, and 120 larvae per 3 ml of food)<br />
out of the six treatments in relation to larval density for both medfly populations used, were<br />
placed into cages containing adult diet, water, and oviposition substrates (domes). Mortality<br />
and female fecundity were recorded daily until death.<br />
Results & Conclusions: The short and the long – lived medfly populations responded<br />
differently to stress induced by larval crowding. Increased larval density (a) reduced larval and<br />
pupal survival only in the case of the short-lived population (Guatemala), and (b) increased<br />
duration of larval development only for the long-lived population (Greece). Pupal size (length<br />
and weight), adult lifespan, and female fecundity were reduced in response to larval density for<br />
both populations used.<br />
Keywords: Ceratitis capitata, intraspecific competition, stress, life-history, fitness
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
AGE AFFECTS THE MATING COMPETITIVENESS OF MALE MEDITERRANEAN<br />
FRUIT FLIES.<br />
Papanastasiou, Stella 1 ; Diamantidis, Alexandros 1 ; Carey, James R. 2 & Papadopoulos,<br />
Nikos, T. 1<br />
1 Laboratory of Entomology and Agricultural Zoology, Department of Agriculture, Crop Production and Rural<br />
Environment, University of Thessaly Greece, Phytokoy St. Nea Ionia (Volos), 38446 Magnisia, Greece; Email:<br />
nikopap@uth.gr; 2 Department of Entomology, University of California, Davis CA 95616<br />
Background: Mating behavior of male Mediterranean fruit flies Ceratitis capitata<br />
(Wiedemann) has been extensively studied the last few decades mostly in the frameworks of<br />
the implementation of the Sterile Insect Technique (SIT). However, the effect of age of males<br />
on mating success remains largely unknown, especially for the older males. When sperm<br />
resources are not exhausted in successive matings, theory predicts that the older males would<br />
be more successful against young males in receiving matings. We used medfly males, a male<br />
lekking species, to test the above hypothesis.<br />
Methods: Experiments were conducted during 2007 – 2008 under laboratory conditions. Wildlike<br />
flies, reared in laboratory for 1 – 3 generations were used. We run mating tests under<br />
competitive (two choice) and non competitive (no choice) conditions. In choice tests, 10 young<br />
(12-14 days old), sexually mature, virgin females were brought to choose between 10 control<br />
(sexually mature, non mated, young, 12-14 days old) and 10 older males (15, 20, 30, 40, 50<br />
days old). In no choice tests, the performance of 10 non mated males of different age (10, 15,<br />
20, 30, 40, 50 and 70 days old) to mate with 10 young, sexually mature virgin females was<br />
assessed. In choice tests we recorded the number of copulas of young (control) and older<br />
males. In no choice tests we further recorded the copula duration, the latency time, the amount<br />
of sperm transferred to female spermathecae, and the sperm abundance in testes of mated, and<br />
not mated males.<br />
Results:In choice tests male mating competitiveness dramatically decreased in ages older than<br />
30 days. Fifty days old males achieved only 15% of matings compared to younger – control<br />
males. The proportion of matings decreased in older ages in no choice tests. Nevertheless,<br />
more than 30 and 20% of 50 and 70 days old males achieved copulations respectively. Mean<br />
duration of copula decreased in older males while the latency time to mate increased. Old<br />
males transferred to female spermathecae comparable amounts of spermatozoids with that of<br />
younger males. The number of sperm cells increased in testes of both mated and non mated<br />
males at older ages.<br />
Conclusions: Our results indicate that (I) C. capitata male mating competitiveness decreases in<br />
advanced ages (II) although ageing negatively affects the ability of males to mate (even in no<br />
competitive conditions), older males can still become accepted as mating partners, transferring<br />
comparable amounts of spermatozoids with that of younger males to female spermathecae.<br />
And (III) our data suggest that spermatogenesis occurs in adult males until extreme ages. The<br />
importance of our findings to understand the role of older males for the reproductive behavior<br />
of male lekking species is discussed.<br />
Keywords: Medfly, aging, mating competitiveness, sperm transfer<br />
137
POSTER ABSTRACTS
DATABASES
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
RIESPO: IBERIAN NETWORK ON THE EVALUATION OF EFFICACY AND SIDE<br />
EFFECTS OF CONTROL TREATMENTS AGAINST OLIVE PESTS<br />
Adán, Angeles 1 ; Bento, Albino A. 2 ; Budia, Flor 1 ; Campos, Mercedes 3 ; Cotés, Belén 3 ;<br />
González, Manuel 4 ; Jerez, Carlos 5 ; Paredes, Daniel 3 ; Nave, Anabela 6 ; Medina, Pilar 1 ;<br />
Pascual, Felipe 5 ; Pascual, Susana* 4 ; Pereira, José A. 2 ; Porcel, Mario 3 ; Rei, Fernando 7 ;<br />
Ros, J. Pedro 4 ; Ruano, Francisca 5 ; Ruíz, Manuel 8 ; Sánchez-Ramos, Ismael 4 ; Santos,<br />
Sónia A.P. 2 ; Seris, Elena 4 ; Torres, Laura 6 & Viñuela, Elisa 1<br />
1 Departamento de Producción Vegetal: Botánica y Protección Vegetal. Escuela Técnica Superior de Ingenieros<br />
Agrónomos (ETSIA), Universidad Politécnica de Madrid, Madrid, Spain. 2 Centro de Investigação da Montanha<br />
(CIMO), Escola Superior Agrária, Instituto Politécnico de Bragança, Bragança, Portugal. 3 Estación Experimental<br />
de Zaidín, Consejo Superior de Investigaciones Científicas (CSIC), Granada, Spain. 4 Departamento de Protección<br />
Vegetal, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). Carretera de A Coruña<br />
km 7,5 28040 Madrid, Spain. Email: pascual@inia.es. 5 Departamento de Biología Animal, Facultad de Ciencias,<br />
Universidad de Granada, Granada, Spain. 6 Departamento de Agronomia. Universidade de Trás-os-Montes e Alto<br />
Douro, Vila Real, Portugal; 7 Departamento de Fitotecnia/ICAAM, Universidade de Évora, Évora, Portugal;<br />
8 Laboratorio de Producción y Sanidad Vegetal, Consejería de Agricultura y Pesca, Junta de Andalucía, Jaén,<br />
Spain.<br />
The Iberian network on efficacy and side effects evaluation of control treatments against olive<br />
pests (RIESPO) was constituted in 2009 and funded by a Complementary Action (AC2009-<br />
00045-00-00) of the National Institute of Agricultural and Food Research and Technology<br />
(Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, INIA). The main<br />
objectives of this network is unifying and standardize methodologies applied in field trials,<br />
from the setting up of the assays in the field to the sampling methods used to characterize the<br />
arthropod community of olive groves and the statistical methods used for data analysis.<br />
Currently, the network consists of researchers and technicians from eight centres in Spain and<br />
Portugal. Members participate in research projects focused on pest control in olive groves,<br />
paying special attention to the key pest of this crop, the olive fruit fly (Bactrocera oleae). This<br />
communication aims to present the network RIESPO to those researchers who may be<br />
interested in its activities.<br />
Keywords: Bactrocera oleae, Field trials, Control methods, Efficacy and side effects assessment<br />
143
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
NEW PROGRESS IN TEPHRITID WORKERS NETWORKING<br />
Bakri, Abdeljelil* 1 ; Hendrichs, Jorge²; Reyes, Jesus²; Liedo, Pablo 3 & Robinson, Alan²<br />
1 Insecte Contrôle, Marrakech 40080, Morocco (bakri@ucam.ac.ma). ²Insect Pest Control Section, Joint<br />
FAO/IAEA Programme Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency,<br />
Vienna, Austria. 3 El Colegio de la Frontera Sur, ECOSUR, Tapachula, Chiapas, Mexico.<br />
Tephritid Workers Database (TWD) was established in 2004, and since then the number of its<br />
member keeps growing to reach 1046. To ensure excellence and relevance in networking<br />
among tephritid workers, TWD intends to evolve to achieve this mission. In 2009, TWD knew<br />
two major developments, the establishment of the Tephritid Workers of Western Hemisphere<br />
Database (TWWH) and the reinstatement of the Fruit Fly News (FFN) newsletters.<br />
It took almost one year of hard work to establish the TWWH since the decision was taken at<br />
the 7th Meeting of the Working Group on Fruit Flies of the Western Hemisphere in Mazatlan,<br />
Mexico on November 2008. At present, TWWH is the largest regional group, including 425 of<br />
tephritid workers from 21 countries. The TWWH database is bringing together scientists from<br />
major universities, research institutes and agriculture agencies in the Americas. Their field of<br />
expertise is extending from ecological theory and laboratory investigation to area-wide<br />
programs using the latest technology to control fruit flies.<br />
Fruit Fly News (FFN) was initially produced in hardcopy form in the 1980’s, and used to be<br />
the communication tool of the IOBC Global Working Group on Fruit Flies of Economic<br />
Importance. A total of 11 annual or semiannual issues were produced until 1992. Now, 17<br />
years later, FFN is back and is provided online from the issue FFN 12. The goal of FFN is to<br />
become the communication tool of the worldwide groups working on tephritid fruit flies,<br />
providing timely information on management of tephritid fruit flies, research activities,<br />
publications, ongoing action programs, upcoming events, job opportunities, and news. As a<br />
community means of communication, efforts are made to ensure its sustainability and regular<br />
frequency of delivery. Up to now, there are three issues a year. All contributions and<br />
suggestions are welcome and highly appreciated.<br />
Keywords: Database, newsletter, tephritid, fruit flies, pest
Session 1<br />
Biology, Ecology & Behaviour
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
SEXUAL COMPATIBILITY AMONG POPULATIONS OF ANASTREPHA FRATERCULUS<br />
(WIEDEMANN, 1981) (DIPTERA: TEPHRITIDAE) OF DIFFERENT REGIONS OF<br />
PERU.<br />
Alburqueque, Mónica*; García, Juliana & Quintanilla, Luis.<br />
Desarrollo de Métodos, Unidad de Centros de Producción de Moscas de la Fruta-La Molina (UCPMF) Del<br />
Servicio Nacional de Sanidad Agraria (SENASA), La Molina 1915 Lima, Perú Email:<br />
malburqueque@senasa.gob.pe.<br />
Background: An essential requirement for the application of the Sterile Insect Technique (SIT)<br />
is the evaluation of sexual behavior of wild flies to predict the expected performance of<br />
laboratory colonies under field conditions, the aim of this study was evaluated the sexual<br />
behavior of mass-reared flies and wild populations of Anastrepha fraterculus from six regions<br />
of Peru where the SIT could be applied.<br />
Methods: We evaluate the sexual compatibility among populations of the Peruvian Coast:<br />
Piura, Lambayeque, La Libertad, Ancash, Lima, Ica and mass-reared flies, six field cages<br />
installed on plants of Persea americana Mill were used; the wild flies were 16 days old and<br />
laboratory flies 9 days old. 30 females and 30 males from each population (two at a time) were<br />
released in each cage, in four replicates and couples from a single population as witnesses were<br />
released in the other ones (two cages). After each test the crosses: females of the 2 nd population<br />
with males of the 1 st population and females of the 1 st population with male of the 2 nd<br />
population were carried to laboratory and reproduce them in host fruits.<br />
Results: The results showed that populations were sexually active showing more activity<br />
around noon with some differences in the starting time, 9:00 and 10:00 am and peaks of sexual<br />
activity at 11:00, 12:00 and 13:00. Nevertheless the index of sexual isolation (ISI) ranging<br />
from -0.07 to 0.28, reflecting sexual compatibility between the populations and the mass-reared<br />
strain, indicating that wild individuals successfully mate with each other and with the<br />
laboratory strain. Also the male relative performance index (MRPI) and insect females (FRPI)<br />
ranging from -0.26 to 0.36 reflecting both sexes of insects produced in the laboratory is as<br />
effective as the wild to mate.<br />
Conclusions: Our results of behavior, it is confirmed that the colony used for mass rearing of<br />
sterile insects for the implementation of sterile insect technique for pest control is compatible<br />
with the flies from the regions studied. Efforts should be initiated to determine if populations of<br />
other natural mountain and forest regions in the same group and the feasibility of applying the<br />
sterile insect in these regions.<br />
Keywords: Anastrepha fraterculus, Tephritidae, SIT, sexual compatibility, Peru.<br />
147
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SUSCEPTIBILITY OF SECHIUM EDULE TO FRUIT FLIES (DIPTERA: TEPHRITIDAE) IN<br />
LABORATORY CONDITIONS<br />
Atiama, Toulassi * ; Deguine, Jean-Philippe; Douraguia, Elisabeth & Quilici, Serge<br />
Centre de coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), Département<br />
des Systèmes Biologiques, Saint-Pierre (La Réunion), France. E-mail: toulassi.atiama@cirad.fr<br />
Background: Sechium edule (chayote) is a climbing cucurbit largely cultivated under arbour in<br />
La Réunion but its production has strongly declined in recent years. Farmers attribute the yield<br />
losses to fruit flies (Tephritidae, Dacini) (Bactrocera cucurbitae, Dacus ciliatus and Dacus<br />
demmerezi), accordingly current studies are carried out in order to evaluate in the field the<br />
damage of fruit flies. Our study aimed to assess in laboratory the susceptibility of S. edule to<br />
fruit flies (B. cucurbitae, D. ciliatus, D. demmerezi), particularly in studying (i) the future of<br />
eggs laid naturally in fruit by gravid females, and (ii) the development of larvae artificially<br />
deposited in fruit.<br />
Methods: Two types of experiments were conducted in 2009 and three types of fruit were<br />
compared: S. edule (young fruit 10cm) and C. pepo as a control. In<br />
the first trial, for each species of flies, fruit (S. edule and C. pepo) were individually exposed to<br />
cohorts including gravid females and the number of pupae obtained and adult flies emerged<br />
from each fruit were then counted after two weeks. In a second trial, pieces of fruit were<br />
artificially infested by L1 larvae of each species, and their survival rate was measured over six<br />
days.<br />
Results: Development of the three species was better in C. pepo than in S. edule, for B.<br />
cucurbitae (a mean of 615 pupae per fruit of C. pepo versus 0 pupae in S. edule), for D.<br />
demmerezi (445 versus 250 pupae), and for D. ciliatus (15 versus 10 pupae). The mean number<br />
of B. cucurbitae pupae obtained per fruit of C. pepo was not significantly different (P>0.05)<br />
from that of D. demmerezi. Complete development (eggs to adult flies) on S. edule was<br />
observed only for D. demmerezi and D. ciliatus. However, D. demmerezi showed better larval<br />
survival and emergence rate on S. edule than D. ciliatus. For both species, larval development<br />
was better in younger fruits than in mature ones.<br />
Conclusions: These results provide new information on the development of fruit flies in S.<br />
edule. The low levels of infestation observed and the difficulties of development for the larvae<br />
suggest that fruit flies might not be the only cause of yield losses in chayote crops. These<br />
results are important to provide guidelines and to adapt the fruit flies management scheme to<br />
the particular case of S. edule crops.<br />
Keywords: Sechium edule, fruit flies, larval development, emergence rate, pupae
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
HOW DO ALTERNATIVE HOST FRUIT TREES INCREASE CERATITIS CAPITATA<br />
(WIEDEMANN) POPULATIONS IN CITRUS CLEMENTINE GROVES?<br />
Campos, José M.*; Martínez-Ferrer, María T. & Fibla, José M.<br />
IRTA-Amposta. Ctra. de Balada, Km 1. E- 43870 Amposta (Tarragona) Spain. E-mail: jmiguel.campos@irta.es<br />
Background: Ceratitis capitata is a polyphagous specie that causes great losses in citrus crops.<br />
The presence of alternative host fruit trees has been proved to lead the Medfly dynamics in<br />
citrus groves. Frequently, home gardens and also isolated host trees are close to early<br />
Clementine varieties that ripen during the same period. Thus, sanitary measures are promoted<br />
to prevent medfly populations to migrate to citrus orchards.<br />
Methods: During 4 years the influence of alternative host fruit trees was studied in Clementine<br />
citrus orchards. From 6 to 7 Tephri-Trap lured with Tri-Pack, plus DDVP as a toxicant were<br />
placed along the citrus orchard. The number of adults caught per trap was recorded weekly.<br />
Fruit maturity was determined by measuring the rind colour index with a Minolta CR-400®<br />
chromameter. In addition, during 2 years, 274 samples of 14 different fruit species were<br />
collected weekly from all the citrus growing area and placed in containers at 25º C where<br />
medfly larvae emerged were counted. Fruits were counted and weighted, thus the number of<br />
larvae per fruit and kg of fruit was quantified.<br />
Results: All the sampled fruit species, both cultivated and wild were attacked by medfly. Figs<br />
and jujube fruits produced the highest amount of medfly larvae, over 100 larvae per kilo. The<br />
sanitary measures on stone fruit orchards (basically peaches) surrounding citrus had a strong<br />
influence on medfly populations, but did not threaten the harvest since did not coincide with<br />
citrus ripening. Medfly adults from overripe fruits in orchard borders moved and achieved<br />
distances as shown by captures in traps. Medfly control by mass-trapping in those fruit trees<br />
was not enough to prevent the migration of insects to the citrus crop, since captures on the<br />
border of the orchard were high.<br />
Conclusions: Both fig and jujube trees that grow in the nearness of citrus crops, produce the<br />
highest medfly populations when early Clementine varieties are susceptible to be attacked by<br />
this pests. Consequently, enhancing mass trapping technique in areas is required even when<br />
pest control in alternative host fruit trees is performed.<br />
Keywords: Medfly, Ceratitis capitata, Clementine, alternative host trees, traps.<br />
149
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PERFORMANCE OF AUGMENTORIUM AS A SANITATION TECHNIQUE AGAINST<br />
FRUIT FLIES (DIPTERA: TEPHRITIDAE) IN REUNION ISLAND<br />
Deguine, Jean-Philippe*; Atiama, Toulassi; Douraguia, Elisabeth; Jacquard, Cathy;<br />
Simiand, Christophe; Delatte, Hélène & Quilici, Serge<br />
CIRAD (Centre de Coopération Internationale en Recherche Agronomique pour le Développement), Département<br />
des Systèmes Biologiques, Saint-Pierre (La Réunion), France ; E-mail: jean-philippe.deguine@cirad.fr<br />
Background. Tephritid fruit flies cause severe damage to fruit and vegetable crops in Reunion<br />
Island. Instead of the curative approach to reduce existing populations, the first step proposed<br />
for their management is sanitation. This method is based on an original technique firstly<br />
developed by USDA in Hawaii utilizing a tent-like structure called an “augmentorium” which<br />
aims to sequester adult flies emerging from infested fruit while allowing the parasitoids to<br />
escape, via a net placed at the top of the structure. This study focused on the performance and<br />
the efficiency of the augmentorium prototype recently tested in Reunion Island and particularly<br />
(i) the number of adult flies that can potentially be sequestered in an augmentorium in the field;<br />
(ii) the efficiency of the net mesh for fly sequestration and parasitoid escape; (iii) the feasibility<br />
of producing compost with infested fruit collected in the field.<br />
Method. The potential number of flies that could be sequestered was estimated by measuring in<br />
the lab the emergence of several species of flies from infested fruit collected in the field from<br />
2005 to 2009 in different sites of the island. Emergence of adult flies was measured for six<br />
species of flies: (i) Bactrocera cucurbitae, Dacus ciliatus and D. demmerezi attacking three<br />
species of Cucurbits (pumpkin: Cucurbita maxima; cucumber: Cucumis sativus and courgette:<br />
Cucurbita pepo); (ii) Bactrocera zonata, Ceratitis rosa and C. capitata attacking one species of<br />
fruit (mango: Mangifera indica). The sequestration of three of these fly species (B. cucurbitae,<br />
B. zonata and C. capitata) and the escape of two of their parasitoids (Psyttalia fletcheri and<br />
Fopius arisanus) were assessed in the Cirad laboratory in Saint-Pierre in 2008. Preliminary<br />
tests on the feasibility of producing compost were then conducted in Saint-Pierre in 2009,<br />
mixing courgette and other components.<br />
Results. Collections of infested fruits showed the following means of emerged adults per kg of<br />
fruit: 76 for mango (B. zonata, C. rosa, C. capitata); 217 for cucumber, 340 for pumpkin and<br />
594 for courgette (B. cucurbitae, D. ciliatus, D. demmerezi). The efficiency of the mesh chosen<br />
for the prototype of augmentorium (hole area 1.96 mm²) proved to be perfectly effective in the<br />
lab with 100% of sequestration of adult flies. In the same way, 100% of the parasitoids were<br />
able to escape from the mesh if they choose to do so. In addition, we showed that a ratio of<br />
50:30:20 of courgette, sugar cane stem and chicken litter respectively was well adapted to<br />
produce compost.<br />
Conclusion. These results confirm the relevance and the efficiency of the augmentorium in an<br />
agroecological crop protection. As a sanitation technique against fruit flies, the augmentorium<br />
sequesters on average several hundreds of adult flies per kg of infested fruit. As a biological<br />
control method, it may contribute to increase parasitoid populations which are often low<br />
because of the previous and significant pesticide pressure. The augmentorium can also be<br />
considered as a useful tool to produce compost in the context of sustainable agriculture. The<br />
technique of sanitation using the augmentorium is now well accepted by farmers in pilot areas<br />
in Reunion Island.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
IMPACT OF FRUIT FLIES (DIPTERA: TEPHRITIDAE) ON CHAYOTE (SECHIUM<br />
EDULE) IN REUNION ISLAND.<br />
Deguine, Jean-Philippe*; Atiama, Toulassi; Douraguia, Elisabeth; Gilles, Benoît;<br />
François, Thomas & Quilici, Serge.<br />
CIRAD (Centre de Coopération Internationale en Recherche Agronomique pour le Développement), Département<br />
des Systèmes Biologiques, Saint-Pierre (La Réunion), France; E-mail: jean-philippe.deguine@cirad.fr<br />
Background. Chayote is a popular vegetable in Reunion Island. Traditionally and extensively<br />
cultivated for its leaves, it is nowadays usually intensively cultivated under treillises for the<br />
production of fruit. A high number of fruit fallen on the ground result in a significant loss in<br />
production. Farmers attribute this to three fruit fly species (Bactrocera cucurbitae, Dacus<br />
ciliatus and Dacus demmerezi) which roost in the trellises. This situation provokes major<br />
insecticide applications in the chayote trellises to combat fruit flies. The present study aimed<br />
therefore at evaluating the real incidence of fruit flies in chayote cultivation.<br />
Methods. The experiments were conducted in 2008 and 2009 and observations in trellised<br />
crops were performed in different sites on the island. In 2008, hundreds of fruit were collected<br />
at a single site to study the susceptible stage of the chayote fruit while the development of<br />
samples of 20 recently blemished fruit and 20 unblemished fruit was regularly followed in the<br />
field. In addition, 120 blemished fruit were collected in 2008 in two sites 20 fruit were weekly<br />
collected at a site in 2009 and the emergence of adult flies was measured in the Cirad<br />
laboratory over 15 days. In addition, bird nets were placed under the trellis of three fields in<br />
2009 to collect fallen fruit and the number of blemishes and the infestation of the fruit gathered<br />
were examined. In the lab, eight chayote fruit were exposed to gravid females per fly species<br />
and dissected.<br />
Results. In situ, fruit smaller than 6 cm length appeared to be unaffected by female blemishes.<br />
Two to four weeks after blemishes under the trellis, the blemished fruit did not show any<br />
morphological difference compared to unblemished fruit. This observation was strengthened<br />
by the results of emergence tests: only one of 120 blemished fruit collected in 2008 and only<br />
five fruit collected from 219 fruit harvested in the crop in 2009 showed emergence of flies. D.<br />
ciliatus appeared to be the only species able to emerge not only from these infested fruit<br />
collected in the field but also from the fruit exposed to gravid females in the lab. A defence<br />
reaction of the fruit to eggs or L1 larvae of B. cucurbitae and D. demmerezi was also observed<br />
by dissecting infested fruit in the lab. Finally, other field observations showed that only 13 of<br />
197 fruit fallen from the trellis were infested by larvae, meaning that 93 % of fruit fell for other<br />
reasons.<br />
Conclusion. This study provides first results on the real incidence of fruit flies in chayote crops<br />
in Reunion Island. Although fly adults roost in chayote trellis, these pests are not responsible<br />
for the significant production losses recorded by the farmers. This suggests that the chemical<br />
protection used against fruit flies in chayote trellis is not appropriate, especially when one<br />
considers the secondary negative effects of the chemical treatments. A physiological shedding<br />
caused by an overloaded production of fruit under this type of intensive cultivation under trellis<br />
could be the main cause of the fruit fall and this requires further research.<br />
Keywords: fruit flies, chayote, Sechium edule, Reunion Island<br />
151
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
INFERENCES ON COMPETITIVE INTERACTIONS FROM DISTRIBUTION PATTERNS OF<br />
COEXISTING ANASTREPHA FRATERCULUS AND CERATITIS CAPITATA POPULATIONS.<br />
Devescovi, Francisco 1 ; Segura, Diego F. *1 ; Liendo, María C. 1 ; Bachmann, Guillermo 1 ;<br />
Vera, M. Teresa 2 ; Milla, Fabián H. 1 ; Bouvet, Juan P 3 & Cladera, Jorge L. 1<br />
1 Instituto Nacional de Tecnología Agropecuaria (INTA), Instituto de Genética “Ewald A. Favret” (IGEAF), De<br />
los Reseros y Repetto s/n (1712), Buenos Aires, Argentina; Email: dsegura@cnia.inta.gov.ar; 2 Estación<br />
Experimental Agropecuaria (EEA) INTA Concordia, Entre Ríos, Argentina; 3 Estación Experimental Agropecuaria<br />
Ovispo Colombres (EEAOC), Tucumán, Argentina.<br />
Background: Anastrepha fraterculus (native to South America) and Ceratitis capitata (exotic<br />
species in this region) have shared host fruit species in Argentina for more than one century.<br />
Little is known about the mechanisms that allow the coexistence of these tephritids. Previous<br />
works suggested that avoidance of infested fruit would act as a stabilizing force minimizing the<br />
negative effects of interspecific competition. We tested here the hypothesis that these species<br />
exploit available resources avoiding larval competition by analyzing the spatial distribution of<br />
infestation, particularly co-occurrence patterns.<br />
Methods: A. fraterculus and C. capitata infestation patterns were evaluated from pupae<br />
obtained from Psidium guajava. Fruit were collected in Northeast region of Argentina<br />
(Concordia) and were processed individually. Frequencies of fruit infested by one, both or<br />
none species were compared with two theoretical distributions, expected under a random<br />
oviposition hypothesis and under an avoidance (of already infested fruits) hypothesis. As an<br />
alternative method we use null models (Ecosim 7.0). Each method was tested at three spatial<br />
scales, for each sampling date.<br />
Results: Both methods showed that infestation patterns varied according to the spatial scale. At<br />
the largest scale there was a tendency towards co-infestation, while at a reduced scale<br />
(particularly at the tree scale), the pattern revealed either a random oviposition behavior or<br />
avoidance of infested fruit. At this scale, a tendency towards avoiding infested fruit occurred<br />
only when fruit infestation levels were moderate.<br />
Conclusions: Our study suggests that competition avoidance behavior could be densitydependent.<br />
This mechanism could, at least partially, permit the coexistence of A. fraterculus<br />
and C. capitata. Other factors, such as environments and host species associated with agroecosystems<br />
could also be playing a role in structuring population densities.<br />
Keywords: Interspecific competition, distribution patterns, EcoSim, spatial and temporal heterogeneity.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
LIFE CYCLE ASSAY AND ESTIMATING SURVIVAL FUNCTIONS OF KONAR FRUIT<br />
FLY, CARPOMYA VESUVIANA COSTAL IN SOUTH OF IRAN.<br />
Farrar, Nasser 1* ; Golestaneh, S.R. 1 ; Mohammadi, M. 2 & Alemansoor, H. 3<br />
1 Agriculture and Natural Resources Research Center of Bushehr Province, Iran; Email: farrar29@yahoo.com; 2 Research<br />
Institutes of Forests and Rangelands; 3 Agriculture and Natural Resources Research Center of Shiraz Province. Iran.<br />
Background: The Konar fruit fly, Carpomya vesuviana Costa (Diptera: Tephritidae) cause high<br />
damage in quality and quantity of (Konar) Ziziphus spp. in south of Iran. The life table is one<br />
of the most important tools in demographic and gerontology research because it is used to<br />
characterize the mortality and survival properties of cohorts and to quantify the actuarial rate of<br />
aging.<br />
Methods: Field and laboratory studies on the Life cycle of Carpomya vesuviana were carried<br />
out for the three consecutive years in south of Iran. The materials of the Tephritid (Eggs,<br />
Larva, Prepupa and Pupa) were collected by weekly sampling in nature and reared in Petri<br />
dishes containing the soft well aerated soil. The Konar fruit fly was reared on three kinds of<br />
Ziziphus fruits (Z. spina-christi, Z. nummularia, Z. mauritiana) as well as an artificial diet to<br />
study the life cycle parameters under laboratory conditions.<br />
Results: The longevity of adults was from 14.6±4.2 to 25.8±6.3 days in different months. The<br />
sex ratio was 1: 1. The pre-oviposition, oviposition and post-oviposition periods lasted 2 to 8, 3<br />
to 35 and 0 to 12 days, respectively. The female lays egg after mating under fruit skin. The<br />
number of eggs/ovipunctures observed in a single fruit is generally 1 to 4 and a maximum of 8.<br />
Females laid averagely 19.1±5 eggs. Eggs-laying in most females (about 72.8%) is carried out<br />
in the first 3 to 7 days of their life. Females have higher fecundity during November and<br />
February and lower fecundity in March. The incubation period was 1-4 days and about 70.4 to<br />
91.9 percent eggs turned out to be viable. The larval period was 6 to 22 days. Larvae had<br />
longer during December (15.5±3.6) and lived short during March (average 8.5±1.6). The<br />
maggots took 1.8 to 5 hours to prepare puparium. Pupal duration was variably different<br />
between 8-320 days. The shortest was 8 days in March April and the longest was 320 days in<br />
September.<br />
Conclusions: The insect completed 8 to 10 overlapping generations each year. The shortest<br />
time for a complete cycle belonged to the flies which laid eggs in March, April and January.<br />
The longest time for a complete cycle was about 330 day which refers to the eggs laid in<br />
September-October. The over wintering and summering was a pupa within soil profile. This<br />
insect has diapause or might not have any diapause at all. Insect attack intensity was estimated<br />
between 30 and 100 percent in randomly inspected fruits.<br />
Keywords: Carpomya vesuviana, Konar, Ziziphus, Life cycle, Iran<br />
153
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
MATING COMPETITIVENESS OF STERILIZED MASS-REARED MALES AND WILD<br />
POPULATIONS OF ANASTREPHA FRATERCULUS (WIEDEMANN, 1981) (DIPTERA:<br />
TEPHRITIDAE).<br />
García, Juliana*; Alburqueque, Mónica & Quintanilla, Luis.<br />
Desarrollo de Métodos, Unidad de Centros de Producción de Moscas de la Fruta-La Molina (UCPMF) Del<br />
Servicio Nacional de Sanidad Agraria (SENASA), La Molina 1915 Lima, Perú Email: jgarciac@senasa.gob.pe<br />
Background: The South American fruit fly Anastrepha fraterculus (Wiedemann 1830), is one<br />
of the most important agriculture pest in Peru. The Sterile Insect Technique (SIT) would be a<br />
powerful no-pollutant method for its direct control which has been used successfully against a<br />
parental specie Ceratitis capitata (Wiedemann 1829) in Peru. The success of SIT depends on<br />
the quality and the ability of sterile males mate with wild females. Competitiveness tests<br />
measure the ability of sterile males to achieve copulations with wild females and the degree of<br />
the sterility of the eggs produced by wild females when wild and sterile males compete to mate<br />
with them. The aim of this study was to evaluate the mating ability of sterilized mass-reared<br />
(two doses 0.07 and 0.08 KGy/hour) A. fraterculus in comparison with wild males from 5<br />
regions of the Peruvian coast: Lima, Ica, Ancash, La Libertad and Lambayeque.<br />
Methods: “competitiveness” cage (field cage) into which 120 insects sterile males were release<br />
along 40 wild males and 40 wild females; “wild control” cage (plexiglas cage 30 cm×30<br />
cm×40cm) into which 25 males insects wild were release along with 25 wild females; “sterile<br />
control” cage (cages with panel 20x15x15cm) into which 25 sterile males were released along<br />
with 25 females fertile insect. Plums were placed into each cage for a period of 8 days in order<br />
to collect the eggs to measure the induced sterility. The fruits hosts were changed daily and the<br />
percent of eggs hatched was evaluated. Fried's competitiveness index (C) was used whose<br />
values range from 1 to 0, values close to zero indicate superior competitiveness of the wild<br />
male. The weight of the pupae were registered in order to correlate the average weight of the<br />
irradiated pupae and the C value<br />
Results: The results obtained in the Fried test (Which measures induced sterility in eggs) with<br />
irradiated pupae with 0.08 KGy/hour showed a competitiveness coefficient ranging from 0.27<br />
to 0.47; on the other hand irradiated pupae with 0.07 KGy/hour showed a competitiveness<br />
coefficient ranging from 0.30 to 0.45. The lowest sexual performance of the sterile males was<br />
with the Ica population (in both doses), while the highest ones was with the La Libertad<br />
population (0.08 KGy/hour) and Ancash (0.07 KGy/hour); this suggests that sterile males<br />
successfully compete with flies from different parts of the Peruvian coast.<br />
Conclusions: Mating competitiveness is an important quality control parameter that affects the<br />
performance of released sterile insects. The results suggest that sterile males successfully<br />
compete with flies from different parts of the Peruvian coast. This paper shows that despite the<br />
strain of mass rearing A. fraterculus used in the test has been reared since 2001 is still<br />
appropriate for the purposes of implementing the SIT.<br />
Keywords: Anastrepha fraterculus, SIT, sexual Competitiveness, sterile male, wild flies.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DISPERSAL DYNAMICS OF AN INVASIVE FRUIT FLY SPECIES, BACTROCERA<br />
INVADENS, ALONG AN ALTITUDINAL GRADIENT.<br />
Geurts, Katrien 1* ; De Meyer, Marc 2 ; Mwatawala, Maulid 3 ; Erbout, Nathalie 1 & Lens,<br />
Luc 1<br />
1 Terrestrial Ecology Unit, Faculty of Sciences, Ghent University, Gent, Belgium; Email:<br />
marc.de.meyer@africamuseum.be; 2 Entomology Section, Royal Museum for Central Africa, Tervuren, Belgium;<br />
3 Department of Crop Science and Production, Sokoine University of Agriculture, Morogoro, Tanzania.<br />
Background: Bactrocera invadens, a fruit fly species originating from Asia, has become an<br />
invasive pest in Africa. This fruit fly causes a lot of damage to crops and threatens fruit export,<br />
leading to considerable economic losses. In Africa it seems to be spreading not only in latitude<br />
but also in altitude. The ongoing spread of B. invadens can be limited by certain barriers:<br />
climatic conditions, host availability and suitability, and interspecific competition with coldtolerant,<br />
Ceratitis rosa, a native species.<br />
Methods: This study tried to determine the boundaries for the extension of the distribution area<br />
of B. invadens by placing traps and collecting fruit along an altitudinal transect (580-1789 m)<br />
in Morogoro, Tanzania from September 2008 until September 2009. Concurrently<br />
environmental variables (temperature, relative humidity and host availability) were measured<br />
along the transect at the different collection sites.<br />
Results: Diversity increased with rising altitude and was determined by host availability and<br />
suitability. B. invadens abundance and infestation declined with rising altitude and was largely<br />
determined by temperature and host availability. Interspecific competition could also play a<br />
part in determining abundance and diversity. In peach B. invadens was dominated in numbers<br />
by C. rosa, but in mango B. invadens dominated all other fruit fly species, even mango<br />
specialist: C. cosyra. There was an annual dispersal (in January and February) of B. invadens<br />
towards higher altitudes in periods with population surplus. Its reproduction in high altitude<br />
fruits such as peach and pear could mean a colonization of this area.<br />
Conclusions: Because of B. invadens’ strong r- and K-selection and its broad climatic and host<br />
range it is possible that this species will continue to spread to high altitude areas and possibly<br />
pose a threat to subtropical areas, even more so considering the influence of climate change.<br />
Keywords: B. invadens, altitude, dispersal, Africa<br />
155
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
TEPHRITIDS IN THE MAIN CITRUS-GROWING AREA IN COSTA RICA (2007 – 2008).<br />
Hernán Camacho, V.*<br />
Retired Professor University of Costa Rica. Adviser in Tico Frut, Costa Rica. P.O. Box 2100 – 3000. Heredia.<br />
Costa Rica. E-mail hcamachov@hotmail.com<br />
Background: In Costa Rica, 27,000 hectares of land currently produce citrus fruits, these being<br />
mainly sweet oranges (Citrus sinensis). There are 4,519,219 trees for monoculture crops<br />
planted in a total of 20,000 hectares. 15,153 hectares are located in the Huetar Zone, which<br />
occupies a significant part of the northern sector of the country. This area is where most of the<br />
nation’s orange crop is raised. These plantations are located on the border with Nicaragua, and<br />
they extend into that country’s territory, making oranges the most important crop in that region.<br />
In this same zone, the Tico Frut Company operates its plants and other facilities. Tico Frut is<br />
Central America’s largest industrial fruit processing enterprise, and the most important fruit<br />
harvest collection and storage center in all of Costa Rica. In order to best ascertain how to<br />
select the most effective and suitable strategies for tephritid control, the diverse tephritid<br />
populations present in the aforementioned region were studied.<br />
Methods: The study was carried out at six orange plantations (Farms No. 9, 12, 13A, Naranya,<br />
Mefisa, and LBA), these having a total joint area of 5,604 hectares, during the harvest periods<br />
of 2007 - 2008. Yellow Multilure Traps that had been baited with Nu Lure were used, with<br />
checks carried out each week in order to collect and identify the insects that were captured.<br />
Simultaneously, a weekly sampling of 60 ripe oranges took place in the same lots where the<br />
traps had been placed. Larvae and pupae were collected on a weekly basis, as their eclosion<br />
was expected, which would make it possible to identify them.<br />
Results: A total of 1093 traps were set at the plantations; in these traps, 9,009 diptera<br />
specimens were collected, of which 69 were Tephritidae, 803 were Lonchaeidae, and 907 were<br />
Ulididae. The number of tephritid orange pests was low. There were only 2 Mexican fruit flies<br />
(Anastrepha ludens) and 2 Medflies (Ceratitis capitata). Other potential orange pest specimens<br />
of the Anastrepha genus were captured. These were 14 A.striata, 33 A.obliqua, 5 A serpentina<br />
and, 2 A fraterculus. None of these specimens were trapped in or on fruits, but rather only in<br />
traps. In the orange crop sampling, 22,920 fruits were analyzed (382 total samples comprised<br />
of 60 oranges each) and from these samples, the following pests were obtained: 1,154<br />
Lonchaeidae, as well as 380 Muscidae, but no Tephritidae or Ulididae flies. An unforeseen<br />
outcome was that the Lonchaeidae Neosilba batesi caught in traps and in the fruit sampling<br />
accounted for the majority of the pests collected. Another unexpected result was the number of<br />
the Ulídidae Xanthacrona bipustulata captured in traps. The largest number and widest<br />
diversity of tephritids were captured in the area of the industrial plant facilities. In 2007, 112<br />
traps were set, and 1,360 diptera were collected, of which 152 were tephritids (6 A ludens, 1 C<br />
capitata, 116 A striata, 18 A obliqua, and 4 A fraterculus), in addition to 85 lonchaeids. In<br />
2008, 75 traps were set, and these caught 383 flies, of which 28 were tephritids (2 A ludens, 2<br />
C capitata, 18 A striata, and 6 A obliqua), but no lonchaeids. This greater diversity was due to<br />
the large quantity of fruit stored at this locale, originating from crops harvested at a number of<br />
sites around the country.<br />
Keywords: Medfly, Mexican Fruit Fly, Anastrepha spp., Lonchaeidae, Ulididae.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DYNAMIC POPULATION OF FRUIT FLIES IN CASSAVA CROPS (Manihot esculenta)<br />
Montes, Sônia M. N. M.* , ¹; Raga, Adalton² & de Souza-Filho, Miguel F.²<br />
¹Agência Paulista de Tecnologia dos Agronegócios-Polo Alta Sorocabana, P.O. Box 298, 19015-970 Presidente<br />
Prudente, SP, Brazil. E-mail: soniamontes@apta.sp.gov.br; ²Instituto Biológico/Laboratório de Entomologia<br />
Econômica, P.O. Box 70, 13001-970 Campinas, SP, Brazil.<br />
Background: Cassava is a subsistence crop, especially for small proprietiers in the north and<br />
NE regions of Brazil. Many insect pests infest cassava crops in Brazil. The damage increases<br />
according to technological level and its intensive production. Tephritidae larvae infest easily<br />
fruits and stems. The purpose of this work was to determine the dynamic population and<br />
identify de fruit fly diversity in a mixed cassava cultivars. The cassava crops is situated in<br />
Presidente Prudente municipality, State of São Paulo, Brazil (UTM 7545288.76 m N,<br />
459930.31 m E and altitude 424.29 m).<br />
Methods: The experiment was conducted from February to May 2005, January 2006 to<br />
December 2007 in cassava field composed by industry and table cultivars `IAC-12´, `IAC-13´,<br />
`IAC-14´, `IAC-15´, `Espeto´, `Branca de Santa Catarina´, `Roxinha´, ´Fécula Branca´, `Fibra´<br />
and `IAC 576-70´. The flies were weekly collected by two yellow McPhail plastic traps at<br />
1.20m high, using 400 mL per trap of 5% hydrolisated protein. Mature cassava fruits were<br />
picked at random for evaluation of fruit fly species incidence. The area was kept free of<br />
insecticide sprays.<br />
Results: We captured a total of 600 flies from the genus Anastrepha (372 males and 228<br />
females), related to: A. montei Lima (152), A. sororcula Zucchi (6), A. pickeli Lima (22) and A.<br />
fraterculus Wied. (2). Based on the flies per trap per day (FTD), the peak reached 1.64<br />
(13/4/2005), 1.43 (12/04 and 17/05/2006) and 2.21 (18/04/2007). The infestation rate in fruits<br />
was 2.0% in `Espeto´ and `IAC 14´. Only A. montei was recovered from cassava fruits.<br />
Conclusions: Low fruit fly diversity was registered in cassava crops by McPhail trapping,<br />
which only A. pickeli and A. montei was related with cassava plants. We obtained low<br />
population in cassava fruits by A. montei.<br />
Keywords: McPhail traps, FTD, Anastrepha, occurrence.<br />
157
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
OCCURRENCE OF FRUIT FLIES IN MANGO ORCHARD IN THE WEST REGION OF<br />
THE STATE OF SÃO PAULO, BRAZIL.<br />
Montes, Sônia M. N. M. * 1 ; Raga, Adalton 2 & de Souza-Filho, Miguel F. 2<br />
¹Agência Paulista de Tecnologia dos Agronegócios-Pólo Regional Alta Sorocabana Rodovia Raposo Tavares<br />
km561, Caixa 298, Presidente Prudente, SP, Brazil, 19015-970. E-mail soniamontes@apta.sp.gov.br; 2 Instituto<br />
Biológico/Laboratório de Entomologia Econômica, P.O. Box 70 Campinas, SP, Brazil 13001-970.<br />
Background: The mango crops are widely distributed in the majority of tropical and<br />
subtropical countries. Fruit flies (Diptera: Tephritidae) are key pests in mango crops in Brazil.<br />
The objective of the present work was evaluate the fruit fly population dynamic and diversity<br />
in an mixed orchard of Haden, Tommy Atkins and Palmer, in Presidente Prudente, SP, Brazil.<br />
Methods: From August 2005 to December 2007, flies were captured weekly in nine yellow<br />
McPhail traps. Inside the trap we disposed 400 mL of 5% hydrolisated protein (Isca<br />
Tecnologias Ltda.), renewed weekly.<br />
Results: We collected a total of 65,956 flies: 2,788 of Anastrepha spp. (4.23%) and 63,168 of<br />
Ceratitis capitata Wied. (95.77%). From the Anastrepha spp. specimens, 55.45% are males<br />
and 44.55% females. For medfly, we captured 43.69% males and 56.31% females. During<br />
2006 we obtained 85.65% of C. capitata and 94.08% of Anastrepha spp. from the captured<br />
flies. The following species were identified: Anastrepha obliqua (Macquart) (91.92%), A.<br />
fraterculus (Wied.) (3.24%), A. sororcula Zucchi (1.37%), A. pseudoparalella (Loew)<br />
(1.11%), A. striata Schiner (0.77%), A. barbiellinii Lima (0.26%), A. daciformis Bezzi<br />
(0.09%), A. haywardi Blanchard (0.09%), A. leptozona Hendel (0.60%), A. zenildae Zucchi<br />
(0.09%) and A. montei Lima (0.85%). Only A. obliqua was considered constant species.<br />
Conclusions: During the monitoring C. capitata was dominant. Among eleven Anastrepha<br />
species, only A. obliqua was constant and dominant. Mango is a common host for Ceratitis<br />
capitata, A. fraterculus and A. obliqua in the State of São Paulo.<br />
Keywords: Anacardiaceae, population dynamic, Tephritidae, faunistic analyses.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
PROLONGED PUPAE DIAPAUSE AFFECTS VITAL FITNESS COMPONENTS OF<br />
RHAGOLETIS CERASI ADULTS.<br />
Moraiti, Cleopatra A. & Papadopoulos, Nikos T.<br />
Laboratory of Entomology and Agricultural Zoology, Department of Agriculture, Crop Production and Rural<br />
Environment, University of Thessaly Greece, Phytokoy St. Nea Ionia (Volos), 58446 Magnisia, Greece. Email:<br />
nikopap@uth.gr.<br />
Background: Diapause is a physiological process allowing insects to exploit seasonally<br />
fluctuating resources, diversify, and colonize harsh environments. Although several<br />
physiological and ecological aspects of diapause have been studied in detail, little is known<br />
regarding the effects of prolonged diapause on adult life history traits. The European cherry<br />
fruit fly, Rhagoletis cerasi L. (Diptera: Tephritidae) is a univoltine species undergoing<br />
obligatory pupae diapause for, usually, one or more years. Thus, it forms an excellent model<br />
organism to test whether prolonged pupae diapause tradeoffs adult fitness components.<br />
Methods: In 2007-2009, we studied the demography of R. cerasi adults of three Greek<br />
populations, emerging from pupae that terminated diapause after one (1 st year) or two (2 nd year,<br />
prolonged diapause) cycles of high-low-high temperatures. The three populations were<br />
originating from: i) Mikra (Thessaloniki), ii) Kamari (Magnisia), and iii) Kernitsa (Achaia). In<br />
all populations, pupae, recovered from field infested, ripe sweet cherries, were placed at 25 ο C<br />
for 2.5 months, then at 3 ο C for 6 months, and finally at 25 ο C until adult emergence (1 st<br />
adults). Overlaying pupae were held at 25 ο C for 2.5 months, and then at 3 ο C for an additional<br />
period of 6 months, and ultimately, returned to 25 ο C until adult emergence (2 nd year adults). A<br />
pair of newly emerged adults (one male and one female) were held into individual cages<br />
containing oviposition substrates (five ceresin domes), standard adult diet and water. Fifty such<br />
pairs (replicates) were established for each population and diapause regime. Daily egg<br />
production and female and male ages at death were recorded under constant laboratory<br />
conditions (25 ± 1 ο C, 65 ± 5% R.H., and L14:D10). Additionally, the size of the body of each<br />
adult (female and male) was assessed at death by measuring thorax length and width, and head<br />
width. Somatometric data showed that both males and females emerging after prolonged pupal<br />
diapause were larger than their counterparts emerging the first year. Average longevity, for<br />
both males and females, differed significantly among populations the first year, but not among<br />
adults emerging after prolonged pupae diapause. Within each population, average longevities<br />
of adults emerging the second year were either similar or longer than of those emerging the<br />
first year. Average fecundity was higher for females emerging the first year than that of their<br />
second year counterparts. Fecundity varied substantially among the different populations the<br />
first year (260 – 380 eggs/female), but it was similar after prolonged pupae diapause (≈100<br />
eggs/female). Oviposition and postoviposition periods of females emerging the second year<br />
were 15 and 5 days shorter and longer, respectively, than that of females emerging the first<br />
year.<br />
Conclusions: Collectively, these data suggest that prolonged pupae diapause: i) is associated<br />
with larger adult body size, ii) decreases lifetime female fecundity rates, iii) alters the age<br />
specific patterns of oviposition, and iv) does not substantially affect the adult lifespan.<br />
Keywords: European cherry fruit fly, life history traits, demographic parameters, body size, fitness cost,<br />
Tephritidae<br />
year<br />
159
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE INFLUENCE OF HOST FRUIT AND TEMPERATURE ON THE BODY SIZE OF<br />
ADULT CERATITIS CAPITATA (WIEDEMANN) IN THE LABORATORY AND IN FIELD<br />
POPULATIONS.<br />
Navarro-Campos, Cristina *1 ; Martínez-Ferrer, María T. 2 ; Alcaide, Juan 1 ; Campos, José<br />
M. 2 ; Bargues, Laura 1 ; Fibla, José M. 2 ; Marzal, Carmen 1 & Garcia-Marí, Ferran 1 .<br />
1 Instituto Agroforestal Mediterráneo (IAM), Universidad Politécnica de Valencia, Camino de Vera s/n, 46022,<br />
Valencia, Spain; Email: crinacam@posgrado.upv.es; 2 IRTA Amposta. Carretera de Balada, Km 1. E- 43870-<br />
Amposta (Tarragona), Spain.<br />
Background: The adult body size of medfly, Ceratitis capitata (Wiedemann) (Diptera:<br />
Tephritidae), varies in natural conditions. Both temperature during larvae development and<br />
host fruit quality have been cited as possible causes for this variation. Body size is an important<br />
fitness indicator in insects; larger individuals usually exhibit increased survival, longevity,<br />
mating success and fecundity. We studied the influence of host fruit and temperature during<br />
larvae development in adult body size in field populations of medfly with the aim to quantify<br />
body size variation throughout the year.<br />
Methods: The size of adult C. capitata was measured and analyzed using wing surface area as<br />
an estimate of adult size. Flies measured had three origins. Firstly, flies periodically collected<br />
along the year in field traps from 32 groves in Spain, during the period 2003-2007. Secondly,<br />
flies evolved from field collected fruits. Finally, flies reared in laboratory on different foods<br />
and in different temperature regimes.<br />
Results: In the lab, wing area of male and female adult medflies varied significantly with<br />
developing temperature of larvae, being larger at the lowest temperature (females: F = 114.06;<br />
df = 4, 209; P < 0.0001, males: F = 107.89; df = 4, 209; P < 0.0001). Adult size was also<br />
significantly different depending on host fruit in which larvae developed (females: F = 11.23;<br />
df = 3; 127; P < 0.0001; males: F = 7.84; df = 3; 131; P = 0.0001). Females developed in<br />
apricot were the largest followed by those developing in peach, plum and orange. The size of<br />
the flies captured at the field varied seasonally showing a gradual pattern of change along the<br />
year. The largest individuals were obtained during winter and early spring and the smallest<br />
during late summer. Size of adult flies showed less variation according to host fruit in which<br />
larvae developed in the field.<br />
Conclusions: Adult size in field populations of C. capitata changes periodically influenced<br />
mainly by temperature, and secondarily by species of host fruit. These observations could<br />
improve our current background on the behavior of C. capitata in the field. The importance of<br />
final adult size and its implication in the sterile insect technique are discussed.<br />
Keywords: medfly, body size, host, temperature-size rule.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FRUIT FLIES OF ECONOMIC IMPORTANCE WITH SPECIAL REFERENCE TO<br />
BACTROCERA CUCURBITAE, A GLOBAL REVIEW.<br />
Nikpay, Amin 1 & Kahrarian, Morteza 2<br />
1 Division of Plant Protection, Sugarcane & By-Product Development Company, Salman Farsi Unit, Ahwaz, Iran;<br />
Email amin_nikpay@yahoo.com; 2 Department of Agriculture, Islamic Azad University, Kermanshah Branch.<br />
Background: Tephritids are one of the most destructive and diversified group of insects. There<br />
are more than 4000 species of fruit flies in the world in 500 genera which the melon fly<br />
Bactarocera cucurbitae (Coquillett) (Diptera: Tephritidae) is an important pest of<br />
Cucurbitaceae (melon, watermelon, pumpkin, squash) in different countries such as most Asian<br />
countries, Middle East and Hawaii. This species is responsible for both direct economic losses<br />
in vegetable production and for the considerable efforts implemented for the detection and<br />
eradication programs in those countries where this pest is found. Also fruit flies of the family<br />
Tephritidae are considered the most important quarantine insect pests in terms of international<br />
trade. Chemical control, first used as the only management strategy, led to insect resistance,<br />
high cost and contamination of fruits and the environment and negative effect on beneficial<br />
arthropods. Due to this reason, in recent years several tactics and strategies such as sterile<br />
insect techniques, bait spray, Biological controls, legislative control and pheromone traps,<br />
alone or in combination have been used. However, in spite of the tremendous investments and<br />
efforts made to date, the pest has not been pushed out of all invaded areas and the war is far<br />
from being won. In this study, we reviewed several control strategies and focus on success or<br />
failure of each method.<br />
Keywords: Melonfly, Bactrocera cucurbitae, Sterile Insect Technique, Pheromones<br />
161
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
TEPHRITIDAE ASSOCIATED WITH MANGO DAMAGES IN THE WEST OF BURKINA<br />
FASO: IDENTIFICATION AND IMPORTANCE OF THE DAMAGES.<br />
Ouédraogo, S. Nafiba* 1 , Vayssières, Jean-François 2 , Dabiré A Rémy 3 & Rouland-Lefèvre<br />
Corinne 4<br />
1 Université Paris Est, 32 Avenue Henry Varagnat 93-140, Bondy, France. osylvainn@yahoo.fr; 2 Centre<br />
International de Coopération et de Recherche Agronomique pour le Développement/ Institut International<br />
d’Agriculture Tropicale (CIRAD/IITA), Cotonou, Bénin; 3 Institut de l’Environnement et de Recherche Agricoles<br />
(INERA), Bobo-Dioulasso, Burkina Faso; 4 Institut de Recherche pour le Développement (IRD) UMR<br />
BIOEMCO/IBIOS, Bondy, France.<br />
Background: Mango fruits export constitutes an important source of currency for Burkina<br />
Faso. However, the damages inflicted by Tephritidae (fruits fly) to mangos threaten the<br />
viability of this sector. Poor knowledge of fruits fly species responsible for these damages<br />
makes difficult the development of an adapted control strategy. The present study aims to<br />
identify the Tephritidae species associated with these damages and to assess their importance<br />
on eight cultivars. Two hypotheses were formulated i) The damages caused by Tephritidae on<br />
the mango in the West of Burkina are caused by several fruits fly species and, ii) The<br />
infestation rate of the mangos by Tephritidae varies significantly depending on the cultivar.<br />
Methodology: Fruits of six grafted cultivars (Amélie, Brooks, Keitt, Kent, Lippens and<br />
Springfield) and two non-grafted ones (Mango vert and Sabre) of mango (Mangifera indica)<br />
served as plant material. During mango seasons 2008 and 2009 (March to August), a sample of<br />
36 fruits per cultivar was taken every 2 weeks from eight orchards. Fruits were incubated using<br />
the method described by Vayssières et al. (2004), so as to determine infestation rates. Pupae<br />
collected from the infested fruits allowed after hatching, identification of fruits fly species<br />
(Diptera: Tephritidae) associated with the damages. The analysis of variance at 5 % threshold<br />
after data transformation allowed comparison of the infestation rates.<br />
Results: Seven species of Tephritidae of the genus Bactrocera (1species) and Ceratitis (6<br />
species) were identified. B. invadens and C. cosyra appeared from most of the infested fruits<br />
(61.14 % and 36.1 % of the hatched adults, respectively). The rates of mango infestation varied<br />
between 2.4 % for Springfield at the beginning of the season (April, 2008) and 69.2 % for<br />
Brooks at the end of the season (August, 2008). The comparison test of the averages has<br />
distinguished five classes of cultivars according to infestation rates (F = 6.181; P = 0.0001).<br />
Amélie was the grafted cultivar most weakly infested whereas Keitt was found to be most<br />
strongly infested.<br />
Conclusions: This study has demonstrated (1) the existence of seven Tephritidae species<br />
infesting the mango among which B. invadens and C. cosyra are the most associated with the<br />
damages and (2) the significant influence of cultivars on the rate of mango infestation by<br />
Tephritidae. These results suggest a much better comprehension of the biology and ecology of<br />
these two species in the study zone for a more effective control strategy of fruits fly damages.<br />
Keywords: Bactrocera invadens, Ceratitis cosyra, Mangifera indica, quarantine insects, infestation rate.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
NATURAL FRUIT FLY INFESTATION IN STONE FRUITS IN THE STATE OF SÃO<br />
PAULO, BRAZIL.<br />
Raga, Adalton* 1 ; Paula, Laís Ívina Silva de²; de Souza-Filho, Miguel F. 3 & Lopes de<br />
Castro, Jairo 4<br />
¹Instituto Biológico, P.O. Box 70, 13001-970, Campinas, SP, Brazil. E-mail: adalton@biologico.sp.gov.br;<br />
²Pontificia Universidade Católica de Campinas, Av. John Boyd Dunlop - s/n°, 13060-904, Campinas – SP, Brazil.<br />
3 Instituto Biológico, P.O. Box 70, 13001-970, Campinas, SP, Brazil. 4 Experiment Station, APTA, P.O. Box 62,<br />
18300-970, Capão Bonito, SP, Brazil.<br />
Background: Fruit fly species (Diptera: Tephritidae) were considered key pests in Rosaceae<br />
crops in Brazil, causing fruit losses and increasing its costs due to cover insecticide sprays. In<br />
the case of peaches, the growers provide additionally fruit bagging after reaching 2.0 cm in<br />
diameter. We report in the study the fruit fly species recovered from peaches, nectarines and<br />
plums in the southwest region of the São Paulo, Brazil.<br />
Methods: The study was conducted in the Germoplasm Bank installed at the Experimental<br />
Station of Agência Paulista de Tecnologia dos Agronegócios, in the municipality of Capão<br />
Bonito. During 2004, 2005 and 2006, mature fruits from 34 varieties were picked up at random<br />
from the canopy of unsprayed trees. The samples were bring to the laboratory and each fruit<br />
were placed on sieved sand in an isolated plastic container covered by a fine mesh cloth. Fruit<br />
fly puparia were extracted from the sand with a sieve and kept in a small cage until adult<br />
emergence.<br />
Results: Except `Harry Pieckstone´, all remainder varieties were infested by Tephritidae:<br />
Àurora 1´, `Flor da Prince´, `Douradão´, `Dourado 1´, `Dourado 2´, `Ouromel 2´, ´Aurora 2´,<br />
`Joia 1´, `Joia 2´, `Joia 4´, `Regis´, `Ouromel 3´, `Centenária´, `Rubrosol´, `Diamante´,<br />
`Rosalina´, `Brasão´, `Josefina´, `Reubennel´, `Januaria ´, `Grancuore´, `Gema de Ouro´, `Roxa<br />
de Itaquera´, `Kelsey 31´, `Kelsey Paulista´, `Tropical´, `Carmesin´, `Fla 680/13´, `Fla 87-7´,<br />
`Fla 87-1´, `Okinawa´ and Mume (Prunus mume). From a total of 1,454 puparia of Tephritidae<br />
emerged 1,328 adults. We obtained only 669 Ceratitis capitata Wied. (361 females and 308<br />
males) and 659 Anastrepha fraterculus Wied. (302 females and 357 males).<br />
Conclusions: Ceratitis capitata and A. fraterculus are the most poliphagous fruit fly species in<br />
the state of São Paulo. Consequentelly, the IPM strategies in stone fruit crops depend on the<br />
knowledge of suitable hosts for both fruit fly species in the local agroecossystem.<br />
Keywords: medfly, south-american fruit fly, Rosaceae; peach, plum<br />
163
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
WOLBACHIA IN THE ORIENTAL FRUIT FLY BACTROCERA DORSALIS (DIPTERA,<br />
TEPHRITIDAE) IN PAKISTAN.<br />
Rasool, Bilal 1,2* ; Schuler, Hannes 2 ; Riegler, Markus 3 ; Arthofer, Wolfgang 4 & Stauffer,<br />
Christian 2<br />
1 Nuclear Institute for Agriculture & Biology, Faisalabad, Pakistan, bilalisb2001@yahoo.com. 2 Institute of Forest<br />
Entomology, Boku, University of Natural Resources & Applied Life Sciences, Vienna, Austria. 3 Centre for Plants<br />
and the Environment, University of Western Sydney, Australia. 4 Molecular Ecology Group, University of<br />
Innsbruck, Austria.<br />
Background: Wolbachia are a common and widespread group of endosymbiontic bacteria<br />
which is capable of manipulating reproduction of arthropod and nematode hosts. This<br />
bacterium causes a variety of reproductive abnormalities including cytoplasmic incompatibility<br />
CI. Wolbachia has been recently reported as a possible means for insect pest population<br />
control. Bactrocera dorsalis is an important economic pest of orchards and vegetables and is<br />
found in Asia including Pakistan. A Wolbachia strain was identified in 19 specimens analyzing<br />
over 1500 Bactrocera dorsalis individuals from China. Here we report on the identification of<br />
a different Wolbachia strain from a Pakistani B. dorsalis population.<br />
Methods: Collection of B. dorsalis adults was done from two regions in Pakistan. DNA was<br />
extracted from individual adult flies. Analysis was done by PCR using general wsp primers,<br />
cloning and sequencing. Individuals without visible amplicon were tested for low titer<br />
infections by Southern blot.<br />
Results: In most B. dorsalis individuals a PCR amplicon was detected using the general wsp<br />
primers. Comparison of the wsp sequences showed differences between Pakistan and China<br />
specimens. Analysing the individuals without visible amplicon by Southern blot revealed a<br />
100% Wolbachia infection rate in Pakistan.<br />
Conclusions: All individuals analyzed revealed to be infected by one Wolbachia strain<br />
contrasting the data of the Chinese population. MLST markers will be applied to this new<br />
Wolbachia strain to bring more insight into its phylogenetic relationships.<br />
Keywords: Bactrocera dorsalis, Wolbachia, Pakistan, phylogeny, wsp
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
COLOR AND VOLATILES RESPONSES OF FEMALE BACTROCERA DORSALIS<br />
(HENDEL) (DIPTERA: TEPHRITIDAE) TO DIFFERENT MANGO RIPENING STAGES.<br />
Rattanapun, Wigunda* 1 ; Amornsak, Weerawan 2 & Clarke, Anthony R. 3<br />
1 Department of Agricultural Technology, Faculty of Technology and Community Development, Thaksin<br />
University, Phatthalung, 93110, Thailand. Email: rwigunda@tsu.ac.th; 2 Department of Entomology, Faculty of<br />
Agriculture, Kasetsart University, Bangkhen Campus, Chatuchak, Bangkok, 10900, Thailand. 3 School of Natural<br />
Resource Sciences and CRC for National Plant Biosecurity, Queensland University of Technology, GPO Box<br />
2434, Brisbane, Qld 4001, Australia.<br />
Background: Tephritid females locate their host plants using both visual and olfactory cues.<br />
The color and volatiles preferences of female fruit flies tend to relate to their host fruits. The<br />
responses of female fruit flies to fruit abundance are partially the consequence of the<br />
stimulation by host fruit volatiles and physiological stage of female flies. Understanding the<br />
visual and olfactory responses of tephritid female flies is important for the development of trap<br />
design, trap deployment and orchard design strategies.<br />
Methods: This study isolated host visual and olfactory cues for three mango ripening stages.<br />
The artificial mangoes of the three mango ripening stages (unripe, ripe and fully-ripe) were<br />
used to study the effect of color to host preference of the Oriental fruit fly, Bactrocera dorsalis<br />
(Hendel) (Diptera: Tephritidae). A Y-tube olfactometer was used to determine the preference<br />
of B. dorsalis female flies on volatiles of each mango ripening stage. A 21-22-day-old, gravid<br />
female B. dorsalis, which had no prior oviposition experience, were used to study on color and<br />
volatile preferences. All experiments were carried out between 07:00-15:00 hours.<br />
Results: Analysis did not detect a significant differential response by female flies to the three<br />
different ripening stages of artificial mango (female response to unripe fruit = 23 flies; ripe<br />
fruit = 31; fully-ripe = 36; χ 2 = 2.867, d.f. = 2, P = 0.239, n = 90). Many female flies were<br />
observed attempting to oviposit in artificial mangoes after arrival at fruit. Female flies showed<br />
a significant preference to the volatiles of fully-ripe mango over the volatiles of ripe mango (χ 2<br />
= 11.267, d.f. = 1, P = 0.001, n = 60) and unripe mangoes (χ 2 = 48.60, d.f. = 1, P < 0.0001, n =<br />
60). When female flies were presented with the volatiles of ripe and unripe mangoes, female<br />
flies significantly preferred the ripe mango volatiles over the unripe mango volatiles (χ 2 =<br />
48.60, d.f. = 1, P < 0.0001, n = 60).<br />
Conclusions: Host fruit color played only a minor role in host quality assessment by B. dorsalis<br />
female flies, whereas host fruit volatiles played an important role in the determination of host<br />
quality. Many female fruit flies tried to oviposit into artificial mango demonstrates that color<br />
and shape of host fruit alone can stimulate oviposition behavior of female fruit flies.<br />
Keywords: artificial mango, Bactrocera dorsalis, host fruit, oviposition, Y-tube.<br />
165
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FRUIT FLY SPECIES (DIPTERA: TEPHRITIDAE) DIVERSITY IN THREE REMNANT<br />
AREAS OF THE HIGLY ENDAGERED ATLANTIC RAINFOREST IN THE STATE OF<br />
BAHIA, BRAZIL.<br />
Silva, Janisete G. 1* ; Santos, Mirian S. 1 ; Navack, Kelly I. 1 & Araujo, Elton L. 2<br />
1 Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz (UESC) Email: jgs10@uol.com.br;<br />
2 Departamento de Ciências Vegetais, Universidade Federal Rural do Semi-Árido, BR 110, km47, Mossoró, RN,<br />
59625-900, Brazil<br />
Background: The Atlantic rainforest in Brazil is considered one of the richest biomes on earth<br />
and southern Bahia harbors one of the few and largest remnants of the highly endangered forest<br />
and an important center of endemism.<br />
Methods: This study was carried out in three remnants of the Atlantic rainforest in the<br />
municipalities of Belmonte, Ituberá, and Una, in the state of Bahia, for two years. We<br />
documented the diversity and dynamics of the three most frequent fruit fly species. Fruit flies<br />
were collected using 10-12 plastic McPhail traps baited with hydrolyzed protein and set up in<br />
tree canopies for each fragment.<br />
Results: A total of 7,470 fruit fly adults was captured (4,376 females and 3,094 males).<br />
Seventeen Anastrepha species were recorded, Anastrepha antunesi Lima, Anastrepha<br />
bahiensis Lima, Anastrepha dissimilis Stone, Anastrepha distincta Greene, Anastrepha<br />
fraterculus (Wied.), Anastrepha furcata Lima, Anastrepha grandis (Macquart), Anastrepha<br />
leptozona Hendel, Anastrepha minensis Lima, Anastrepha obliqua (Macquart), Anastrepha<br />
pickeli Lima, Anastrepha pseudoparallela (Loew), Anastrepha quiinae Lima, Anastrepha<br />
serpentina (Wied.), Anastrepha simulans Zucchi, Anastrepha sororcula Zucchi, and<br />
Anastrepha zenildae Zucchi. Ceratitis capitata (Wied.) specimens were trapped only in the<br />
fragment in Belmonte. A. minensis and A. simulans are reported in the state of Bahia for the<br />
first time. In the fragment in Belmonte, A. fraterculus showed two population peaks in<br />
December 2007 and August 2008, whereas A. leptozona and A. distincta showed a population<br />
peak in April de 2008. In the remnant in Ituberá, A. distincta, A. bahiensis, and A. fraterculus<br />
showed a population increase from March to June 2008, A. distincta showed a population peak<br />
in April and A. bahiensis and A. fraterculus in May. In the fragment in Una, few specimens<br />
were trapped during the study period and A. fraterculus showed a population peak in February<br />
and April 2008, A. distincta and A. antunesi showed a small population increase in April 2008.<br />
In the three fragments, the capture of flies was restricted to a few months during the study<br />
period coinciding with the host fruiting period.<br />
Conclusions: This is the first fruit fly survey in the Atlantic rainforest in Bahia. Our results<br />
elevated to 33 the number of Anastrepha species registered in the state. Most Anastrepha<br />
species recorded were monophagous or oligophagous.<br />
Financial support: FAPESB, Plantações Michelin da Bahia, CNPq and UESC.<br />
Keywords: Anastrepha, population fluctuation, Atlantic rainforest.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FRUIT FLIES (DIPTERA: TEPHRITIDAE) AND PARASITOIDS ASSOCIATED TO<br />
SPONDIAS MOMBIM L. IN UNA, BAHIA, BRAZIL.<br />
Silva, N. M. O. 1 ; Araujo, Elton L. 2 ; Silva, Janisete G. 1*<br />
1 Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz (UESC), Rodovia Ilhéus-Itabuna km<br />
16, Ilhéus, Bahia, 45650-000, Brazil. Email: niveamosilva@yahoo.com.br; 2 Departamento de Ciências Vegetais,<br />
Universidade Federal Rural do Semi-Árido, BR 110, km47, Mossoró, RN, 59625-900, Brazil<br />
Background: Yellow mombim (Spondias mombim L.) fruit have a very characteristic flavor<br />
and are greatly appreciated in Brazil. In southern Bahia, the collection of this fruit has been a<br />
source of income for local producers who produce frozen concentrate juice for the domestic<br />
market. There are no studies regarding levels of infestation by fruit flies or control methods for<br />
this Anacardiaceae species in southern Bahia, which may be attributed to the fact that the fruits<br />
are still obtained in an extractive way. This study aimed at identifying the fruit fly species and<br />
associated parasitoids that use yellow mombim (‘cajá’) as a host.<br />
Methods: We carried out a survey from June 2005 to June 2007 collecting fruit samples weekly<br />
from tree canopies and from the ground on an experimental 1 ha orchard in Una, southern<br />
Bahia, Brazil The fruit were individually placed in plastic vials covered with cloth to avoid<br />
reinfestation until adults and parasitoids emerged. We determined infestation rates, pupal<br />
viability, emergence rates, parasitism rates and fruit loss due to infestation.<br />
Results: A total of 1,003 fruits (13.18 Kg) were collected from which 729 puparia were<br />
obtained. A total of 331 tephritid adults and 183 parasitoids emerged, representing a pupal<br />
viability of 70.50%. The mean infestation rate was 0.73 puparia/fruit and 55.3 puparia/Kg of<br />
fruit. The mean emergence rate was 45.4%. Only two Anastrepha species were recovered from<br />
the infested fruit, Anastrepha obliqua and Anastrepha antunesi. A. obliqua showed a 78%<br />
emergence rate and A. antunesi showed a 22% emergence rate. The parasitism rate recorded<br />
was 35.6%. Three braconid species emerged from Anastrepha larvae/puparia, Doryctobracon<br />
areolatus and Utetes anastrephae (Opiinae) and Asobara anastrephae (Alysiinae). Parasitoids<br />
in the subfamily Opiinae showed a frequency of 84% and thes only species in the subfamily<br />
Alyssinae showed a frequency of 16 %. D. areolatus was the most frequent parasitoid<br />
associated to A. obliqua and A. antunesi. Fruit loss ranged from 11% to 93.4%. Infestation rates<br />
over 87.54 puparia/Kg of fruit corresponded to more than 50% of fruit loss.<br />
Conclusions: Our results indicate that A. obliqua and A. antunesi damage yellow mombim fruit<br />
during the period of highest fruit production and that any action directed at controlling these<br />
fruit flies in the field should be applied prior to or during this period.<br />
Financial support: from CNPq, FAPESB, and UESC.<br />
Keywords: Anastrepha, Spondias mombim, parasitoids.<br />
167
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE CURRENT KNOWLEDGE ON ANASTREPHA SCHINER (DIP., TEPHRITIDAE) IN<br />
THE STATE OF AMAPÁ, BRAZIL.<br />
Silva, Ricardo A. *1 ; Deus, Ezequiel G. 1 ; Pereira, Júlia D. B. 2 & Zucchi, Roberto A. 3<br />
1 Empresa Brasileira de Pesquisa Agropecuária (Embrapa Amapá), Rodovia JK, km 5, n. 2600, CEP 68903-419,<br />
Macapá, Amapá, Brazil. E-mail: adaime@cpafap.embrapa.br; 2 Universidade Federal do Amapá (UNIFAP),<br />
Macapá, Amapá, Brazil. 3 Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ), Universidade de São Paulo<br />
(USP), Piracicaba, São Paulo, Brazil.<br />
Background: Until the 1990s, studies with fruit flies in the state of Amapá located in northern<br />
Brazil were virtually nonexistent. However, studies were intensified after the official detection<br />
of the carambola fruit fly (Bactrocera carambolae) in Oiapoque border with French Guiana in<br />
1996. B. carambolae is a quarantine pest for Brazil and is restricted to the state of Amapá,<br />
under official control of the Brazilian government. This study aims to list the species of fruit<br />
flies of Amapá, highlighting those that have no known host.<br />
Methods: In recent years, periodic and intensive surveys of fruits (cultivated and wild species)<br />
have been conducted in almost all counties of Amapá, generating a significant amount of<br />
information on the diversity of fruit flies and host plants.<br />
Results: Currently 32 species of the genus Anastrepha are recorded for Amapá. Of these, 17<br />
species are known hosts: A. anomala, A. antunesi, A. atrigona, A. bahiensis, A. coronilli, A.<br />
distincta, A. fraterculus, A. hastata, A. leptozona, A. obliqua, A. parishi, A. pseudanomala, A.<br />
serpentina, A. sororcula, A. striata, A. turpiniae and A. zenildae. For 15 species there is no host<br />
record in the state of Amapá by having been captured only in McPhail traps: A. amita, A.<br />
binodosa, A. dissimilis, A. duckei, A. flavipennis, A. furcata, A. limae, A. minensis, A. mixta, A.<br />
pickeli, A. pseudoparalella, A. shanonni, A. sodalis, A. submunda and Anastrepha aff.<br />
mucronota. The diversity of tephritid species probably is related to the diversity of fruit species<br />
in the state. On the other hand, little is known about the influence of meteorological factors on<br />
tephritid populations in the region.<br />
Conclusion: Despite reasonable knowledge about the Tephritidae diversity in the state of<br />
Amapá, the knowledge about host plants and natural enemies is still scarce, especially studies<br />
on biology and population ecology.<br />
Keywords: Fruit fly, Bactrocera, diversity, survey, Amazon.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
AMAZON NETWORK FOR RESEARCH ON FRUIT FLIES.<br />
Silva, Ricardo A. *1 ; Pereira, Júlia D. B. 2 ; Deus, Ezequiel G. 1 ; Godoy, Maria J. S. 3 &<br />
Zucchi, Roberto A. 4<br />
1 Empresa Brasileira de Pesquisa Agropecuária (Embrapa Amapá), Rodovia JK, km 5, n. 2600, CEP 68903-419,<br />
Macapá, Amapá, Brazil. E-mail: adaime@cpafap.embrapa.br; 2 Universidade Federal do Amapá (UNIFAP),<br />
Macapá, Amapá, Brazil. 3 Ministério da Agricultura, Pecuária e Abastecimento (MAPA), Departamento de<br />
Proteção de Plantas, Brasília, Distrito Federal, Brazil. 4 Escola Superior de Agricultura "Luiz de Queiroz"<br />
(ESALQ), Universidade de São Paulo (USP), Piracicaba, São Paulo, Brazil.<br />
Background: Little is known about the diversity of fruit flies species (Diptera: Tephritidae);<br />
reported as one of the main pests affecting agriculture worldwide, in addition to with the low<br />
number of researchers working on this biological group in the Amazon region, it motivated the<br />
structuring of a research project in the Amazon to allow the integration between institutions<br />
and generation of information on the families Tephritidae and Lonchaeidae. With funding from<br />
the Brazilian Agricultural Research Corporation – Embrapa was create the "Amazon Network<br />
for Research on fruit flies" led by Embrapa Amapá (August/2007 to July/2010). Bringing<br />
together researchers from research institutions in 10 Brazilian states, the Network aimed to<br />
generate and spread information on the diversity, distribution, host plants and natural enemies<br />
of fruit flies in the Amazon, with emphasis on the carambola fruit fly (Bactrocera<br />
carambolae), quarantine pest, restricted to the state of Amapa and under official control of the<br />
Ministry of Agriculture, Livestock and Supply. The research priorities on B. carambolae were<br />
established by the Scientific Committee of the National Eradication Program of carambola fruit<br />
fly, aiming at to support the actions in the state of Amapá.<br />
Methods: Samplings of fruits and surveys with McPhail traps in various states of the Brazilian<br />
Amazon were performed. In the laboratory, fruits were processed according to the<br />
methodology used to obtain fruit flies.<br />
Results: As results of this project were obtained new records of fruit flies (Tephritidae) and<br />
"lance flies" (Lonchaeidae) species, host plants and parasitoids in the Brazilian Amazon and<br />
Brazil, generating a significant advance in scientific knowledge on these flies. Team members<br />
were also trained mainly in the taxonomy of fruit flies. New research projects were approved in<br />
the network, emphasizing knowledge of fruit flies species of economic importance.<br />
Conclusion: The greater impact of the scientific network will be obtained at the end of 2010<br />
with the book "Fruit flies in Brazilian Amazon: diversity, hosts and natural enemies”, which<br />
will present the innovative results added to the knowledge previously available in the literature.<br />
Keywords: Tephritidae, Lonchaeidae, Bactrocera carambolae, diversity, host.<br />
169
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CUCURBITACEAE HOSTS FOR NATURAL INFESTATION OF ANASTREPHA GRANDIS<br />
(MACQUART) IN THE OF SÃO PAULO STATE, BRAZIL.<br />
de Souza-Filho, Miguel F.* 1 ; Raga, Adalton 1 ; Montes, Sônia M. N. M. 2 ; Azevedo-Filho,<br />
Joaquim A. de 3 ; Garcia, Maria J. De M. 4 ; Almeida, Aparecida M. de 4 & Matioli, André<br />
L. 1 .<br />
1 Instituto Biológico, Caixa Postal 70, CEP 13012-970, Campinas, São Paulo (Brazil). Email:<br />
miguelf@biologico.sp.gov.br; 2 APTA Regional, Pólo Alta Sorocabana, Presidente Prudente, São Paulo (Brazil);<br />
3 APTA Regional, Pólo Leste Paulista, Monte Alegre do Sul, São Paulo (Brazil); 4 APTA Regional, Pólo Centro<br />
Oeste, Bauru, São Paulo (Brazil).<br />
Background: Anastrepha grandis (Macquart) is the most important pest of cucurbits in Brazil<br />
due to the direct damages and restrictions for its exportation. Growers from the western region<br />
of the State of São Paulo export squash under the System Risk Mitigation for A. grandis. The<br />
objective of the present study was investigated the infestation of A. grandis in cucurbits in the<br />
remainder regions of the State of São Paulo.<br />
Methods: From April 2009 to March 2010, 153 cucurbit fruit samples were collected from 22<br />
municipalities. The samples were brought to the laboratory, weighed and kept into plastic box<br />
containing a thin layer of vermiculite and covered with cotton cloth. The fruits were kept for<br />
about 30 days. The pupa were transferred to the glass cup (500 cc) with vermiculite under<br />
transparent glass jars (6000 cc) and kept at 25°C±2°C and 70±10% relative humidity until<br />
adult emergence.<br />
Results: Only six municipalities have showed cucurbit fruits infested by A. grandis: Bauru,<br />
Campinas, Ituverava, Monte Alegre do Sul, Platina and Socorro. The following species and<br />
respective cultivars showed infestation by A. grandis (pupae/fruit; pupae/kg of fruit): Cucumis<br />
sativus ‘Caipira’ (14.5; 29.2), Cucurbita maxima 'Exposição' (55.9; 40.0), Cucurbita moschata<br />
'Caravela' (84.7; 21.4), Cucurbita moschata 'Menina Brasileira' (38.4; 17.7), Cucurbita<br />
moschata 'Canhão' (5.0;0.9), Cucurbita pepo 'Caserta’ (29.5; 46.6) and hybrid C. moschata x<br />
C. maxima 'Tetsukabuto'. No infestation was registered on C. moschata 'Paulistinha'; C.<br />
moschata 'Goianinha'; Cucurbita mixta ‘Caipira’; Lagenaria siceraria and Cucumis sp.<br />
Conclusions: Cucumber (Cucumis sativus), squash (Cucurbita maxima and Cucurbita<br />
moschata), pumpkin (Cucurbita pepo) and the hybrid squash (C. moschata x C. maxima) are<br />
suitable hosts for A. grandis in the State of São Paulo, Brazil.<br />
Keywords: South American Cucurbit Fruit Fly, Cucumis, Cucurbita, Lagenaria, infestation.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FRUIT FLY ON CHILLI IN THAILAND: SPECIES AND ITS BIOLOGY.<br />
Srikachar, Sunyanee*; Plodkornburee, Wipada & Jamroenma, Kriengkai<br />
Entomology and Zoology Group, Plant Protection Research and Development Office, Department of Agriculture,<br />
Bangkok, Thailand. Email: nutaa2000@yahoo.com<br />
Background: Fruit fly (Diptera: Tephritidae) is the major insect pests of chilli cultivation in<br />
Thailand. The Solanum fruit fly, Bactrocera latifrons (Hendel) is known as an insect pest of<br />
Solanaceae crop. This species is distributed throughout South-East Asia, Taiwan, India and southern<br />
China. We studied on species and biology of fruit flies on chilli plantation.<br />
Methods: Study on fruit fly species and its biology on chilli were conducted both in the laboratory<br />
and field conditions during 2005-2008. Monitoring and collection fruit fly species on chilli<br />
were carried out in Nakornpatom, Kanchanaburi, Bangkok, Ubon-Ratchathani, Khon-Kaen and<br />
Chaiyaphum provinces during December 2005 - August 2008. The damaged chilli fruits were<br />
collected and kept in the laboratory until fruit flies emerged for identification. The conditions<br />
of laboratory were 23.95+0.82 �C and 90.24+2.635% relative humidity (RH).<br />
Results: Remarkably, we found only the Solanum fruit fly, B. latifrons at the number of 4,551<br />
individuals with sex ratio 1:1 from the total number of 19,779 infested chilli. Biological study<br />
of B. latifrons was conducted in the laboratory during 2005-2008. Adult females laid their eggs<br />
singly after her preoviposition period of eight days. Egg stage lasted 44-68 hours with the<br />
average of 192.17+75.18 eggs/female. Percentage of egg hatching was 88%. Larval<br />
development period was three instars. Total larval development period was ranged 8-10 days<br />
and averaged 8.76+0.71 days. Pupal period averaged 11.97+0.77 days with the range of 11-14<br />
days. The longevity of adult female and male were averaged 147.90+29.03 and 131.50+12.79<br />
days, respectively. Life table study of B. latifrons in bush red pepper revealed that the<br />
appearance mortality of the first instar was remarkably high up to 31.82% but the third instar<br />
was low to 10.20%, whereas the survival rate was decreased but stage and age were increased<br />
Conclusion: The results revealed that the Solanum fruit fly, B. latifrons is the only one fruit fly<br />
species infested on chilli in Thailand. Life cycle of B. latifrons was 23.56+0.98 days and thhe<br />
survival rate of egg to adult was 33% under laboratory conditions. Egg parasitoid, Forpius<br />
arisanus (Sonan) (Hymenoptera: Braconidae) and larval parasitoid, Diachasmimorpha<br />
longicaudata (Ashmead) (Hymenoptera: Braconidae) were found to be the natural enemies of<br />
B. latifrons.<br />
Keyword: biology, Solanum fruit fly [Bactrocera latifrons (Hendel)].<br />
171
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
POPULATION OF CHILI FRUIT FLY AND ITS DAILY ACTIVITY IN CHILI PEPPER<br />
(CAPSICUM ANNUM) CROP IN LEMBANG WEST JAVA - INDONESIA.<br />
Syamsudin, Tati Suryati 1 ; Arief, M. Ardelia 1 ; Susanto, A. 1,2 & Setyawati, W. 3<br />
1 School of Life Sciences and Technology – Institut Teknologi Bandung, Ganesa no 10 Bandung 40132. Indonesia.<br />
Email: tati@sith.itb.ac.id; 2 Faculty of Agriculture, Pajajaran University, jalan Raya Jatinangor, Bandung<br />
Indonesia; 3 Center for Vegetables Research, Department of Agriculture, Jalan Raya Lembang, Indonesia.<br />
Background: Chili fruit fly is one kind of pest which attack chili pepper fruit (Capsicum<br />
annum) which has economic impact. However, in Indonesia, the species of Bactrocera dorsalis<br />
(complex) always present on the same localities. Many control measure has been apply to<br />
control the population such as application of attractant. More than twenty sibling species are<br />
attracted to methyl eugenol. In order to control Chili fruit fly, we need the information on the<br />
species and its behavior. The objectives of this research are to identify fruit fly species from<br />
chili pepper, daily activity and its damages in the field.<br />
Methods: Chili fruit fly collection was done from chili invested fruit and by methyl eugenol<br />
trap at Lembang, West Java - Indonesia. Morphological character was measured to confirm<br />
species identification such as: male & female genital organ of fruit fly from infested fruit and<br />
from trap caught. Daily activity of fruit fly in the field was observed every hour using methyl<br />
eugenol trap. Fruit fly damage on chili pepper was observed in 20 chili plants in the field.<br />
Results & Conclusions: From invested fruit, the length of aculeus was about 1.9 - 2.19 (2.02 ±<br />
0.09) micron, aedeagus length was about 3219 – 3853 (3534.7 ± 192.9) micron. However the<br />
length of male fruit fly aedeagus caught from the trap was about 3195 – 3975 (3658.3 ± 156)<br />
micron. It was concluded that fruit fly species from chili pepper was Bactrocera papayae.<br />
Male’s fruit fly respond to methyl eugenol trap in the field showed significant activity at 08.00<br />
to 15.00 and the highest activity was at 11.00 to 14.00. From 20 chili plants, almost 100% of<br />
chili plant was damage by chili fruit fly and the intensity of damages for each plant varied<br />
between 4-94%.<br />
Keywords: B papayae, Capsicum annum, fruits damage, morphological character.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
COMPARISON STUDY OF VALATILES PRODUCED BY CALLING MALE OF CERATITIS<br />
CAPITATA FROM DIFERENT POPULATIONS.<br />
Vaníčková, Lucie* 1,2 ; Do Nascimento, Ruth R. 3 ; Júnior, Cyro R. C. 4 ; Kalinová, Blanka 1<br />
& Hoskovec, Michal 1 .<br />
1 Infochemicals Research Group, Institute of Organic Chemistry and Biochemistry of the Academy of Science of<br />
the Czech Republic, Flemingovo nám. 2, CZ-166 10 Praha, Czech Republic; Email: lucielisboa@gmail.com;<br />
2 Department of Chemistry of Natural Compounds, Institute of Chemical Technology in Prague, Praha, Czech<br />
Republic; 3 Instituto de Química e Biotecnologia, Universidade Federal de Alagoas, Maceió, Brazil; 4 Faculdade de<br />
Nutrição, Universidade Federal de Alagoas, Maceió, Brazil.<br />
Background: Ceratitis capitata is one of the most worldspread member from the fruit fly<br />
family (Tephritidae). There have been numerous SIT (Sterile Insect Techniques) programs<br />
running all around the world to control natural populations of this pest. In these programs,<br />
males originating from mass produced laboratory colony are sterilized by irradiation and<br />
released into the field. It is supposed that sterile males which mate with wild females generate<br />
no progeny. The efficency of these programs is based on the assumption that laboratory males<br />
reared on artificial diet are able to compete with wild males. In fruit flies, mating behavior is<br />
quite complex and involves chemical, visual and acoustic communication. Chemical<br />
communication is based on sex pheromone released by males to attract females.<br />
Methods: In order to provide adequate studies concerning compatibility or incompatibility of<br />
laboratory and wild populations of C. capitata, we analyzed and compared the chemical<br />
composition of volatiles released by calling males of one laboratory and two wild populations<br />
of C. capitata using GC×GC/TOFMS technique and Principal Component Analyses (PCA).<br />
Results: Our results indicate that there is a strong correlation between laboratory and wild<br />
populations of C. capitata studied, suggesting that males from these populations use the same<br />
chemical language to attract females.<br />
Conclusions: These results are of significance for the successful implementation of the SIT<br />
program.<br />
Keywords: Medfly, pheromone, populations, SIT, GC×GC/TOFMS, PCA<br />
173
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EMBRYONIC DEVELOPMENT OF ANASTREPHA LUDENS (LOEW).<br />
Zepeda-Cisneros, Cristina S. * ; García-Martínez, Víctor & De León-Crisóstomo, Ángel H.<br />
National Fruit Fly Campaign. Moscafrut Program. Acuerdo SAGARPA-IICA. Central Poniente No. 14 Col.<br />
Centro Tapachula Chiapas, México. Email: cczepeda@hotmail.com<br />
Background: The egg stage and the embryonic development of Anastrepha ludens is one of the<br />
less studied despite the great importance for mass rearing facilities where it is necessary obtain<br />
high percentages of egg hatch. Peculiar mechanisms of elimination of yolk during gastrulation<br />
and final head lobules development has been described for other Anastrepha species. However<br />
A. ludens has not been studied previously. We describe the different egg stages and time during<br />
embryonic development and the emergency process. This knowledge contributes to the<br />
developmental embryonic of this species and is useful in early evaluation of egg hatch, which<br />
is a valuable date to calculate egg density and at the moment of seeding.<br />
Methods: Samples of eggs of a colony maintained in the laboratory during generations, named<br />
“Chiapas”, were collected in oviposition panels with water during 30 minutes; they were<br />
incubated in petri dishes with a humid filter paper and maintained in a bioclimatic chamber at<br />
26 o C. Observations with a transmitted light stereomicroscopy were made each two hours. It<br />
was not necessary to remove the chorion, because the egg covers are transparent. It was<br />
identified the anterior and posterior poles and registered the moment when appear o disappear<br />
the yolk masses. It was observed the movements of larvae near the emergence and identified<br />
the place where it hatches.<br />
Results: The morphological changes of the egg were monitored during the embryonic<br />
development. Based in yolk masses extruded during the embryonic development of A. ludens<br />
four types of eggs were identified: Type 1 without mass yolk, Type 2 only anterior mass yolk,<br />
Type 3 only posterior mass yolk and, Type 4 anterior and posterior mass yolk. The class 3 and<br />
4 were the most abundant, and corroborated that type 1 and 2 presented embryonic<br />
development. It is possible to predict at the third day after oviposition the egg hatch, with<br />
reliability of 99%, because the unfertilized eggs are clearly identified. Near the emergence the<br />
larvae twist 180 o and the majority hatch near the posterior pole.<br />
Conclusions: The egg stages during the embryonic development permit to estimate the<br />
maximum egg hatch one day before seeding.<br />
Keywords: mexfly, extrusion of yolk mass, Anastrepha ludens, embryogenesis, egg hatch, fertility egg
Session 2<br />
Morphology & Taxonomy
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DNA BARCODING AS IDENTIFICATION TOOL IN FRUIT FLY<br />
INTERCEPTION AND SURVEYING ACTIVITIES<br />
Breman, Floris 1 ; Jordaens, Kurt 1 , De Meyer, Marc 2 & Virgilio, Massimiliano 2-3<br />
1 Joint Experimental Molecular Unit (JEMU), Brussels & Tervuren, Belgium. 2 Royal Museum for Central Africa,<br />
Entomology Section, Tervuren, Belgium. 3 Royal Belgian Institute of Natural Sciences, Invertebrates Section,<br />
Brussels, Belgium<br />
Background: DNA barcoding uses the sequences of a particular part of the mitochondrial gene<br />
COI as a unique identifier for all zoological species. DNA barcodes can provide a valuable<br />
alternative to morphological identification in cases where morphological characters do not<br />
provide diagnostic features. Complete taxon coverage is currently not feasible because of the<br />
unavailability of specimens for the rarer species. However, for quarantine interception, and<br />
field survey and monitoring activities with para-pheromone traps, identification of fruit flies<br />
through DNA barcodes could be considered. This activity fits within the TBI (Tephritid<br />
Barcoding Initiative) and the flagship project on Diptera of JEMU<br />
Methods: Our study focuses on the African fauna of the main fruit fly genera of economic<br />
importance (EI): Ceratitis, Dacus, Bactrocera and to a lesser extent Capparimyia and<br />
Trirhithrum. A DNA barcode library will be established for all EI taxa within these genera, as<br />
well as for all taxa regularly encountered in para-pheromone trapping activities in different<br />
parts of the continent.<br />
Separate sets of intercepted fruit fly samples of European NPPO’s and material recently<br />
collected during surveying and monitoring activities in different parts of Africa will be<br />
requested. This material will be identified independently based upon morphological characters<br />
by a taxonomic specialist, and upon DNA barcodes. The efficiency and reliability of both<br />
methodologies will be compared.<br />
Results: Currently, a DNA library has been compiled for more than 120 African taxa. This<br />
includes all major EI species belonging to the target taxa, several congeneric species, and<br />
outliers of related genera. For widespread species, representatives of different geographical<br />
regions were included in order to cover for intraspecific variability. The library also includes<br />
more than 75% of all taxa regularly trapped in para pheromone traps in the last 20 years. As<br />
test material, samples were obtained from French and UK NPPO’s, resulting from fruit fly<br />
interception in fruit consignments from the last five years. In addition, recent material from<br />
survey programs in western and southern Africa was also obtained. Testing is ongoing but<br />
preliminary results show that DNA barcodes can provide an alternative as identification tool,<br />
especially with regard to juvenile stages like larvae found in intercepted fruit consignments.<br />
Difficulties in species differentiation are mainly encountered in recognized species complexes<br />
such as the Ceratitis FAR complex (C. fasciventris, C. anonae, C. rosa).<br />
Keywords: DNA barcoding, taxonomy, Africa<br />
177
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
COULD BE ANASTREPHA OBLIQUA (MACQUART) A COMPLEX OF CRYPTIC<br />
SPECIES?<br />
Canal, Nelson A.*; Castañeda, Maria del R. & Osorio, Armando<br />
Universidad del Tolima, Facultad de Ingeniería Agronómica. Altos de Santa Helena, Ibagué, Tolima, Colombia.<br />
E-mail nacanal@ut.edu.co<br />
Background: Anastrepha obliqua (Diptera, Tephritidae) is an important pest in Neotropical<br />
countries, specially damaging mangoes. This species belong to the fraterculus group, within<br />
one a complex of cryptic species had been described, especially in the A. fraterculus<br />
(Wiedemann) s.l. Resolution of cryptic species status is important for management tactics like<br />
SIT or other advising the pest biology. An artificial rearing of A. obliqua has been performed<br />
for last 10 years in the “Universidad del Tolima” and two populations were collected from<br />
fruits of hobo (Spondias mombin) and Mango (Manguifera indica) in May 2009. Specimens<br />
from mango were reared easily following our protocol; however specimens from hobo can not<br />
be rearing under laboratory conditions, suggesting a difference in the biology.<br />
Methods: Mature fruits from hobo (H), mangoes (M) and plum (P) (Spondias purpurea) were<br />
collected in Prado, Espinal and Guamo (respectively), three neighboring municipalities in the<br />
Department of Tolima, Colombia. Previous fruit flies collect from these hosts had been always<br />
identified as A. obliqua. Adults flies were obtained and third instar larvae from them were<br />
taken from artificial diet when specimens were collected from mango or plum, and from<br />
papaya for specimens collected from hobo. Measurements of adults (thorax, wings and<br />
aculeus) and larvae (long, depth of mandible and long of anterior and posterior spiracles) were<br />
done. Number of tubules in the anterior spiracles and hairs in the posterior spiracles were<br />
observed too.<br />
Results: Aculeus is longer and thinner in H population (1.54±0.04 mm long, 0.088±0.007 mm<br />
wide VS 1.53±0.08, 0.097±0.006 (M) and 1.50±0.01, 0.096±0.006 (P)); however the aculeus<br />
tip was shorter in H (0.182±0.01 mm VS 0.197±0.006 (M) and 0.189±0.01 (P)). The serrate<br />
part was shorter but no wider in the H populations (0.130±0.096mm long, 0.076±0.007mm<br />
wide VS 0.139±0.00, 0.076±0.00 (M) and 0.144±0.01, 0.078±0.005 (P)). The mandible in the<br />
larvae was less depth in H population (0.035±0.01mm) than in other populations (0.080±0.009<br />
(M), 0.076±0.00 (P)) and conspicuous longer (0.272±0.007mm VS 0.265±0.01 (M),<br />
0.261±0.007 (P)); then, the mandibles ratio is so different (0.128±0.04 (H), 0.304±0.04 (M)<br />
and 0.290±0.008 (P)). The anterior spiracles have 11-12 tubules in the H populations and 14-16<br />
in the M and P populations. The posterior spiracles have fewer hairs in the H specimens than in<br />
the other two populations (approximately a half).<br />
Conclusions: Morphological differences and the biological rearing behavior of specimens<br />
collected from hobo suggest two different species (cryptic species) in the A. obliqua s.l. Other<br />
studies like karyotipes, molecular or crossing will help to clear these findings and experiments<br />
had been performed and started in our University.<br />
Keywords: Morphology, adults, larvae, West Indian Fruit Fly, taxonomy.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
SPECIES ANASTREPHA SCHINER (DIPTERA: TEPHRITIDAE) “FRUIT FLIES” AND<br />
GEOGRAPHICAL DISTRIBUTION IN PERU.<br />
Nolazco, Norma*.<br />
Servicio Nacional de Sanidad Agraria (SENASA), Área de Desarrollo de Métodos (DM). Unidad del Centro de<br />
Producción de Moscas de la Fruta (UCPMF). Área de Taxonomía y Sistemática de Moscas de la Fruta. Unidad del<br />
Centro de Diagnóstico de Sanidad Vegetal (UCDSV). Av. La Molina 1915, La Molina; Lima (Perú). Email:<br />
nnolazco@senasa.gob.pe<br />
Background: The growing economic importance of the losses caused by fruit flies has been<br />
ongoing concern and justified the Ministry of Agriculture through the National Service of<br />
Agrarian Health (SENASA). In Peru, where he performed work for the control, suppression or<br />
eradication of fruit flies, you need a reliable practice of identifying, measuring and evaluation<br />
of the populations of Anastrepha, to demostrate the presence or absence pest and thus obtain<br />
the results for the implementation of Integrated Control work their decline. The main objective<br />
of this study was: to know the species of fruit flies present, captured in Mc Phail traps and their<br />
geographical distribution in Peru, carrying out this analysis based on morphological<br />
characteristics of male and female genitalia.<br />
Methods: The specimens were captured in Mc Phail Traps baited weekly during 2003 January-<br />
2009 December, installed in 19 Departments, the flies were preserved in alcohol 70% in sealed<br />
bottles labeled and sent monthly to the entomology laboratory for taxonomic identification to<br />
species level by analysis of male and female genitalia, which in some specimens were<br />
performed for assemblies. For the determination of species it was used the Identification<br />
Manual for fruit flies prepared by Korytkowski in 2007.<br />
Results: The results obtained were distributed in 29 species, in alphabetical order these are as<br />
follows: Anastrepha alveata, A. atrox, A. barnesi, A. chiclayae, A. curitis, A. dissimilis, A.<br />
distincta, A. fraterculus, A. flavipennis, A. grandis, A. hamata, A. kuhlmanni, A. lambda, A.<br />
lanceolate, A. leptozona, A. macrura, A. manihoti, A. montei, A. nigripalpis, A. obliqua, A.<br />
ornata, A. pickeli, A. pseudoparallela, A. schultzi, A. serpentina, A. shannoni, A. striata, A.<br />
tecta and A. willei, being the most widely distributed species and predominance A. fraterculus,<br />
considered the most important species, having been found in abundant populations in almost all<br />
ecological zones of the region. Of all specimens tested, 51.2% were females and 48, 8% were<br />
males, it had a sex ratio of 1:1 approximately.<br />
Conclusions: Anastrepha species captured in Mc Phail traps in Peru is 29, but the most<br />
common are: Anastrepha fraterculus, A. distincta, A. serpentina, A. striata and A. obliqua. The<br />
most widely distributed species are Anastrepha fraterculus and Anastrepha distincta and lower<br />
distribution is A. willei and A. tecta. Departments that had great diversity were Cajamarca,<br />
Junín, Tumbes, Lambayeque and Huánuco.<br />
Keywords: Anastrepha, McPhail Traps, species, fruit flies.<br />
179
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EGG MORPHOLOGY OF ANASTREPHA SPP (DIPTERA: TEPHRITIDAE) IN THE<br />
FRATERCULUS GROUP.<br />
Ronchi-Teles, Beatriz 1 ; De Siqueira Dutra, Vivian 1 ; Gomes Silva, Janisete 2 & Steck, Gary<br />
J. 3<br />
1 Coordenação de Pesquisas em Entomologia, Instituto Nacional de Pesquisas da Amazônia, C. P. 478, 69011-970<br />
Manaus, AM, Brazil. ronchi@inpa.gov.br; 2 Departamento de Ciências Biológicas, Universidade Estadual de Santa<br />
Cruz, Rodovia Ilhéus/Itabuna km 16, Ilhéus, BA, 45650-000, Brazil; 3 Florida Department of Agriculture and<br />
Consumer Services, Division of Plant Industry,Gainesville, FL 32614-7100.<br />
Background: The fraterculus group is the largest and most widespread species group<br />
comprising 29 species, 17 of which occur in Brazil. The need to explore additional<br />
morphological characters, such as those present in eggs and larvae, besides molecular ones, to<br />
better understand the phylogenetic relationships among Anastrepha species has been pointed<br />
out. Nevertheless, there is a paucity of information on egg morphology and relatively few<br />
studies have described the eggs of only 25 species of Anastrepha. This paper describes the egg<br />
morphology of Anastrepha antunesi Lima, 1938, Anastrepha bahiensis Lima, 1937,<br />
Anastrepha coronilli Carrejo & González, 1993, Anastrepha distincta Greene, 1934,<br />
Anastrepha turpiniae Stone, 1942, and Anastrepha zenildae Zucchi, 1979 using SEM.<br />
Methods: The length and the width measures of the entire egg were taken with an ocular<br />
micrometer and a Wild M3C stereoscopic microscope at Instituto Nacional de Pesquisas da<br />
Amazônia – INPA, Manaus, Brazil. In preparation for SEM, eggs were transferred to silicone<br />
capsules and dehydrated in an ethanol series, then critical point dried in CO2 for 3 hours, and<br />
sputter-coated with a gold layer. The eggs were examined in a LEO 1450VP scanning electron<br />
microscope at Museu Paraense Emílio Goeldi, Belém, Brazil and under an optical microscope<br />
(Leica M165C) at INPA. Twenty to thirty five eggs per female of at least three females of each<br />
species studied were prepared for light microscopy. SEM was used to examine the chorion in at<br />
least 10 eggs.<br />
Results: This SEM study on eggs of six species of Anastrepha shows basic similarities to other<br />
species whose eggs have already been described as well as some differences previously<br />
unreported. Anastrepha coronilli and A. distincta belong into the category of eggs with a<br />
smooth or faintly reticulated chorion – the reticulation in eggs of A. antunesi, A. bahiensis and<br />
A. distincta is very faint whereas it is absent in eggs of A. coronilli. Anastrepha turpiniae and<br />
A. zenildae are in the category of those species with a sculptured chorion and both species<br />
show a very pronounced reticulation.<br />
Conclusions: The results presented here broaden our knowledge of eggshell morphology in<br />
Anastrepha species in the fraterculus group, however more detailed work is needed on<br />
additional species to answer important questions regarding phylogenetic relationships within<br />
the species groups and the genus as a whole. We expect our results to increase our knowledge<br />
of the morphology of Anastrepha eggs in the fraterculus group and to be useful for taxonomic<br />
and systematic purposes.<br />
Keywords: Morphology, fruit flies, Amazon, Brazil
Session 3<br />
Genetics & Evolution
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
CROSS-AMPLIFIED MICROSATELLITE MARKERS IN THE EUROPEAN CHERRY FLY,<br />
RHAGOLETIS CERASI (DIPTERA: TEPHRITIDAE), REVEAL AN EXTENDED<br />
STRUCTURING OF THE SPECIES NATURAL POPULATIONS IN GREECE.<br />
Augustinos, Antonios 1,2,3 ; Asimakopoulou, Anastasia K. 3 ; Papadopoulos, Nikos T.* 4 &<br />
Bourtzis, Kostas 1<br />
1 Department of Environmental and Natural Resources Management, University of Ioannina, 2 Seferi st., 30100<br />
Agrinio, Greece. 2 Department of Biology, University of Patras, Greece. 3 Department of Biochemistry and<br />
Biotechnology, University of Thessaly, Greece. 4 Department of Agriculture, Crop Production, and Agricultural<br />
Environment, University of Thessaly, Greece.<br />
Background: The European cherry fruit fly, Rhagoletis cerasi (Diptera: Tephritidae), is a major<br />
pest of cherries in Europe and parts of Asia. Despite its big economic significance, there is a<br />
lack of studies on the genetic structure of its natural populations. Knowledge about an insect<br />
pest on molecular, genetic and population levels facilitates the development of environmentally<br />
friendly control methods. In this study we present the development of 13 microsatellite markers<br />
for the European cherry fly through cross-species amplification and their evaluation through<br />
the genotyping of 220 wild flies collected in Greece. Microsatellites constitute very useful tools<br />
for population analyses, since they are highly polymorphic, dispersed in the genome, codominant<br />
inherited Mendelian markers.<br />
Methods: Thirty-nine primer pairs designed for the amplification of microsatellite markers in<br />
other Tephritidae species were evaluated for their ability to amplify specific amplicons with R.<br />
cerasi DNA as template. Those that did so were used for the genotyping of a small sample<br />
consisting of 20 individuals. The primer pairs that proved polymorphic, easily scorable and<br />
gave reproducible results were used for a preliminary analysis of R. cerasi natural populations<br />
in Greece.<br />
Results: Thirteen primer pairs conformed to the aforementioned criteria and were used for the<br />
genotyping of 220 individual flies, collected from 8 different sampling locations. Two to nine<br />
alleles per locus were observed and the vast majority of the markers conformed to Hardy-<br />
Weinberg Equilibrium (HWE) in each sample. Genetic distances were very high, pointing<br />
towards an extensive structuring of R. cerasi natural populations.<br />
Conclusions: Our results indicate that (a) cross-species amplification is a versatile and rapid<br />
tool for developing microsatellite markers in Rhagoletis spp., (b) the microsatellite markers<br />
presented here constitute an important tool for population studies on this pest, and (c) there is<br />
clear structuring of natural European cherry fruit fly populations.<br />
Keywords: Rhagoletis cerasi, Tephritidae, microsatellites, population genetics, cross-species amplification<br />
183
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE ROLE OF ESTERASES IN MALATHION AND LAMBDA-CYHALOTHRIN<br />
RESISTANCE IN THE MEDFLY CERATITIS CAPITATA (WIEDEMANN).<br />
Cervera, Amelia*; Arouri, Rabeh; Couso-Ferrer, Francisco; Castañera, Pedro;<br />
Hernández-Crespo, Pedro & Ortego, Félix<br />
Centro de Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC),<br />
Departamento de Biología Medioambiental, Madrid, Spain. *Email: ame@cib.csic.es.<br />
Background: The insecticide malathion has been widely used in Spain for control of the<br />
Medfly Ceratitis capitata (Diptera: Tephritidae), and as a result, resistance has developed in<br />
field populations. The main resistance mechanism described so far is a point mutation in the<br />
acetylcholinesterase gene (ace2), the target molecule of malathion, although metabolic<br />
resistance mechanisms mediated by esterases may also be involved, as revealed by bioassays<br />
with the synergist DEF. In 2009, malathion was withdrawn from the EU market, being mostly<br />
replaced by the insecticides spinosad and lambda-cyhalothrin for Medfly control in citrus<br />
orchards. We are maintaining in our laboratory a 200-fold malathion-resistant strain, as well as<br />
different isolines carrying homozygous mutations in the aliesterase gene ae7- which has been<br />
previously associated with malathion resistance in Musca domestica and Lucilia cuprina -<br />
although none corresponds to the resistance mutations reported in those species. The<br />
malathion-resistant strain is also 5 times less susceptible to lambda-cyhalothrin than the<br />
susceptible strain, and has been further selected with lambda-cyhalothrin to give a 40-fold<br />
resistant population. Susceptibility in this strain is partially regained with the synergist DEF,<br />
which suggests that esterases could also be playing a role in resistance to lambda-cyhalothrin.<br />
We have analysed therefore the possible involvement of both carboxylesterases and<br />
aliesterases in malathion and lambda-cyhalothrin resistance in C. capitata.<br />
Methods: Carboxylesterase specific activity towards the substrate α-naphtylacetate (αNA) has<br />
been measured spectrophotometrically in abdomen homogenates of single insects. Aliesterase<br />
specific activity towards the model substrate methylthiobutyrate (MTB) has been measured in<br />
the same way. Two malathion-resistant strains carrying the point mutation in ace2, a lambdacyhalothrin<br />
selected strain and a susceptible laboratory strain, as well as isolines carrying<br />
different ae7 mutations have been analysed.<br />
Results: Enzyme assays revealed that esterase activities were altered in some of the resistant<br />
strains and isolines when compared to the susceptible strain. Those populations that showed<br />
differences in carboxylesterase or aliesterase activity were further studied by native acrylamide<br />
gel electrophoresis (PAGE) using the same substrates, and their isoenzyme profiles compared.<br />
Conclusions: The role of esterases in malathion and lambda-cyhalothrin detoxification is<br />
discussed in terms of Medfly control strategies in field conditions.<br />
Keywords: Medfly, insecticide resistance, esterase, aliesterase, malathion, lambda-cyhalothrin
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
GEOGRAPHICAL DISPERSION OF GLY328ALA ACETYLCHOLINESTERASE<br />
MUTATION ASSOCIATED TO MALATHION RESISTANCE IN C. CAPITATA<br />
(WIEDEMANN, 1824).<br />
Couso-Ferrer, Francisco*; Perera, Nathalie; Beroiz, Beatriz; Cervera, Amelia; Ortego,<br />
Félix; Castañera, Pedro & Hernández-Crespo, Pedro.<br />
Centro de Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC),<br />
Departamento de Biología Medioambiental, Madrid, Spain. Email: fcferrer@cib.csic.es<br />
Background: A point mutation in acetylcholinesterase gene (ace2) resulting in a Gly328Ala<br />
substitution has been associated to malathion resistance in the Mediterranean fruit fly Ceratitis<br />
capitata (Wiedemann, 1824). In addition, differences in susceptibility among field populations<br />
were related to the frequency of the treatments. We have developed a simple PCR-RFLP<br />
method useful to detect the resistant allele in field populations, and performed an analysis of<br />
the geographical dispersion of this allele among C. capitata populations collected in Spain and<br />
other countries worldwide.<br />
Methods: Primers allowing a PCR-RFLP method directed to determine the presence of ace2<br />
gene mutation in genomic DNA have been designed. 20 individuals from 38 C. capitata<br />
populations (27 from Spanish and 11 from other countries) were analyzed.<br />
Results: Analysis by PCR-RFLP revealed that the resistant allele is widespread all over Spain,<br />
being present in 25 from 27 Spanish populations analysed, including populations from different<br />
regions such as Comunitat Valenciana, Catalunya, Andalucia, La Rioja, Aragón, Illes Balears<br />
and Islas Canarias. The resistant allele was not detected in other populations collected in<br />
Guatemala, Brazil, South Africa, Israel, Australia, Morocco, Tunis, Greece, Turkey, Portugal<br />
(Madeira Island) and France (La Réunion). The frequency of individuals carrying a resistant<br />
allele (in homocigosis or heterocigosis) was related to malathion susceptibility as determined<br />
by bioassays.<br />
Conclusions: 1. PCR-RFLP is an effective method to detect the presence of the malathion<br />
resistant allele Gly328Ala in field populations of C. capitata. 2. Gly328Ala AChE mutation is<br />
widespread all over Spain, but is not present in populations from other countries. 3. The<br />
frequency of individuals carrying a resistant allele (in homocigosis or heterocigosis) is related<br />
to malathion susceptibility determined by bioassays.<br />
Keywords: Medfly, organophosphorus resistance.<br />
185
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
POPULATION GENETICS OF TWO FRUIT FLIES DAMAGING CUCURBITS IN LA<br />
REUNIÓN: BACTROCERA CUCURBITAE AND DACUS CILIATUS.<br />
Delatte, Hélène*; Jacquard, Cathy; Simiand, Christophe; Deguine, Jean-Philippe &<br />
Quilici, Serge.<br />
CIRAD (Centre de Coopération Internationale en Recherche Agronomique pour le Développement), Département des<br />
Systèmes Biologiques -UMR PVBMT CIRAD / Université de la Réunion, Saint-Pierre (La Réunion), France. Postal address: 7<br />
chemin de l’IRAT, 97410 Saint Pierre, France. E-mail: helene.delatte@cirad.fr.<br />
Background: Bactrocera cucurbitae and Dacus ciliatus are two fruit fly species damaging<br />
various cucurbit crops. Of Asiatic origin, B. cucurbitae has become an invasive pest in many<br />
countries especially on the African continent. Conversely, D. ciliatus, which originates in<br />
Africa has increased its distribution during the last years and is now becoming a pest in<br />
different Asiatic countries. Both species rank among invasive pests and has been introduced in<br />
La Réunion, a French island in the South-Western Indian Ocean over the last 50 years. They<br />
now damage the whole range of cucurbits cultivated in the island and cause important yield<br />
losses. La Reunion is a subtropical island, with high altitudinal gradient (up to 3000 m) and<br />
two seasons (hot and rainy summer, milder and drier winter). Nevertheless, those two fruit fly<br />
species are found in most of the areas, regardless of the season and the altitude. However, they<br />
show some altitudinal preferences: B. cucurbitae is more abundant in the lowlands and D.<br />
ciliatus at medium altitudes. The aim of this work was to 1) characterize the population<br />
structures of B. cucurbitae and Dacus ciliatus, 2) test host races 3) and seasonal or altitude<br />
effects.<br />
Methods: 2500 Individuals of B. cucurbitae were collected from 32 sites at high, medium and<br />
low altitudes during winter and summer on different host plants all over Réunion island. In<br />
order to study the population structure of both species, we developed sets of microsatellite<br />
markers for both species, and genotyped at those populations. Genetic differentiations among<br />
geographical regions were quantified through hierarchical analysis of molecular variance<br />
(ARLEQUIN) and levels of population admixture were quantified through Bayesian clustering<br />
procedures (STRUCTURE, TESS).<br />
Results: We obtained 13 polymorphic and specific markers for each species. Those markers<br />
were used for genotyping both species. No clear structuration was found for both species<br />
according to altitude, season or host plants. Between all sites we had high levels of admixture<br />
and reveal patterns of inter-regional, and inter seasonal gene flow.<br />
Conclusions: The low differentiation found between sites among all populations tested is<br />
coherent with the recent invasion by one population for both pests into La Réunion. The<br />
productions of those 2 sets of molecular markers are a good tool to evaluate small and large<br />
scale diversity of both species, and trace back the invasion routes.<br />
Keywords: population genetic analysis, microsatellites, Dacus ciliatus, Bactrocera cucurbitae.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EXTENSION OF THE PATHWAY ANALYSIS FOR CERATITIS CAPITATA<br />
(WIEDEMANN) IN THE MEDITERRANEAN AREA.<br />
Hammouti, Nasera 1* ; Griebeler, Eva M. 1 ; Barr, Norman 2 ; Mazih, Ahmed 3 & Seitz,<br />
Alfred 1 .<br />
1 Institute for Zoology, Department for Ecology, University of Mainz, Johann-Joachim-Becherweg 13, D-55128<br />
Mainz, Germany. Email: hammouti.nasera@yahoo.com. 2 Center for Plant Health and Technology, Mission<br />
Laboratory, USDA-APHIS, Moore Air Base, Edinburg, TX 78541, USA. 3 Département Protection des Plantes,<br />
Institut Agronomique et Vétérinaire Hassan II, BP 121 Aït Melloul, Agadir, Morocco<br />
Background: The Mediterranean fruit fly (medfly), Ceratitis capitata, is one of the most<br />
destructive agricultural pests worldwide. Once introduced, programs to manage or eradicate<br />
medfly populations can have tremendous economic costs. To avoid the establishment of this<br />
species, non-infested countries (or regions) can apply strict quarantine standards on the import<br />
of fresh fruits produced in infested areas. Recently, a worldwide pathway analysis of medfly<br />
based on mitochondrial DNA sequences (i.e., a portion of the cytochrome oxidase I [COI] gene<br />
and a locus including the NADH-dehydrogenase subunits 5 and 4 [N5N4] genes) has been<br />
published by Barr (2009). This diagnostic tool applies an exclusionary principle to gain<br />
knowledge of probable geographic sources of an introduction, considering six predefined<br />
geographical regions: Mediterranean area, Central America, South America, Hawaii, Australia<br />
and Sub-Saharan Africa.<br />
Methods: In Morocco, the medfly is the most serious pest infesting citrus orchards. In our<br />
study, we sequenced both COI and N5N4 markers from Moroccan populations and used the<br />
data to assign the Moroccan population to one of the six predefined regions using the<br />
exclusionary principle. Moreover, we compare diversity within the country, and update the<br />
reference database of medfly diversity. In a second approach to pathway analysis, we applied a<br />
SAMOVA (Spatial Analysis of MOlecular VAriance, a without a priori method) on the whole<br />
samples (from Barr 2009 and Morocco) to refine the geographical grouping described by the<br />
pathway analysis.<br />
Results: For both markers, the most common haplotype among the medfly populations from<br />
Morocco supports that they are genetically similar to the Mediterranean region. We found four<br />
new haplotypes for COI and eight new haplotypes for N5N4. One COI haplotype is common in<br />
Kenya, a probable source for the Moroccan populations. For the more variable marker N5N4,<br />
the SAMOVA assumes a subdivision of the Mediterranean unit in two groups, a Western one<br />
(Morocco, Spain and Portugal) and an Eastern one (Tunisia, Egypt, Italy, Israel, Greece and<br />
Malta).<br />
Conclusions: We conclude that the generation of additional DNA sequences of N5N4 and COI<br />
genes for individuals from many regions increases the discrimination power of the pathway<br />
analysis tool. The construction of a large reference database from worldwide medfly<br />
populations will contribute to identify potential sources of introduction. Such information can<br />
help to improve quarantine procedures.<br />
Keywords: Medfly, pathway analysis, mitochondrial DNA, SAMOVA, Mediterranean area<br />
187
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IMPROVEMENT OF GENETIC SEXING STRAINS THROUGH THE INDUCTION OF<br />
CHROMOSOMAL INVERSIONS IN ANASTREPHA LUDENS LOEW (DIPTERA:<br />
TEPHRITIDAE).<br />
Ibañez-Palacios, Jorge*; Zepeda-Cisneros, Cristina S. & García-Martínez, Víctor.<br />
Nacional Fruit Fly Campaign. Moscafrut Program. Acuerdo SAGARPA-IICA. Central Poniente No. 14 Col.<br />
Centro, CP. 30700. Tapachula Chiapas, México. Email: jorgeip05@hotmail.com<br />
Background: Genetic Sexing Strains (GSS) has been development to be included in the Sterile<br />
Insect Technique (SIT) programs. These GSS have been constructed on the basis of induced<br />
translocations Y-autosome which allows discriminate and eliminate the females in early<br />
development stages so that only males could be released in the field. Because meiotic<br />
recombination presented in males, the GSS have a certain percentage of insect aberrant product<br />
of recombination between the marker selected for the GSS construction (black pupae bp) and<br />
the break point of translocation, it is needed to develop colonies in which the phenomenon of<br />
recombination is reduced or nullified in its entirety through the induction of chromosomal<br />
inversions. For this, it is necessary to have another phenotypic marker linked to bp mutation;<br />
this new mutant found in 2009 was named “purple iridescence” (im), for what is this current<br />
work.<br />
Methods: To induced and isolate chromosomal inversions irradiated bp males were crossed to<br />
im females. The resultant F1 females were then crossed in single pairs to bp/bp; im/im double<br />
mutant males and the F2 offspring were screened for families showing absence or a low<br />
percentage of recombinant phenotypes. When such families were observed, to produce an<br />
homozygous inverted strain, the bp males were crossed to im females, then the F3 wild type<br />
males and females were inbreeding and finally the homozygous bp insects F4 offspring are in<br />
homozygous condition.<br />
Results: The mutation im is a monogenetic, autosomal recessive gene characterized by the<br />
presence of a bright purple halo in the eyes of the insects easily to identify; these insects also<br />
present pupae more elongated and thinner than the wild type. There have been eight trials from<br />
witch have been obtained five families carrying chromosomal inversions: Family 4, 154, 63, 79<br />
and 120. Families 63 and 79 showing 3.70% and 3.95% of recombinant phenotypes<br />
respectively. The families 4 and 120 presented very weak insects when the chromosomal<br />
inversions were in homozygous condition in the F4 and they could not survive by themselves.<br />
Finally, the family 154 was successfully established as a homozygous colony with 500 insects<br />
and has not been recovered recombinant insects from this colony.<br />
Conclusions: It was established a recombinant suppressor strain for A. ludens using the new<br />
phenotypic marker purple iridescence. This chromosomal inversion will help to reduce the<br />
recombination frequency in genetic sexing strain Tapachula-7.<br />
Keywords: genetic sexing strains, inversion A. ludens
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DEVELOPMENT OF A GENETIC SEXING STRAIN FOR ANASTREPHA LUDENS USING A<br />
BLACK PUPAE (bp + ) TRANSLOCATION IN EGFP/Y-INSERTION MALES.<br />
Meza, J. Salvador 1* , Handler, Alfred M. 2 , & Zepeda-Cisneros, Cristina S. 1<br />
1 National Campaign Fruit Flies. Programa Moscafrut. Acuerdo SAGARPA-IICA, México. Central Poniente No.<br />
14 Col Centro CP 30700 Tapachula Chiapas México. Email: mezajs@hotmail.com; 2 Center for Medical,<br />
Agricultural, and Veterinary Entomology, Agricultural Research Service, USDA, Gainesville FL USA.<br />
Background: In order to improve the sterile insect technique (SIT) several germ-line<br />
transformation systems have been developed in pest insects. The Mexican fruit fly, Anastrepha<br />
ludens, in particular was transformed using piggyBac vector constructs, pXL-PUbEGFP/Asb2t-<br />
DsRed.T3. During this transformation one transgenic line with Y-linked insertion was obtained,<br />
expressing EGFP only in males, although the red fluorescence from the testes was not<br />
observed. In addition, a black pupae mutation (bp) was previously isolated from the mass<br />
rearing MOSCAFRUT facility, and this marker has the potential to create a genetic sexing<br />
strain (GSS-bp) by translocating the wild type allele of the mutation (bp + ) to the Ychromosome.<br />
In a bp - /bp - mutant strain having the Y-bp + translocation, male brown pupae<br />
males can be separated from the mutant black pupae females for genetic sexing.<br />
Methods: To improve the male-specific Y-linked marking (and sex-specific selection) in the<br />
GSS-bp strain, we translocated autosomal bp + allele onto the Y-EGFP chromosome to create<br />
Y-EGFP/bp + . Pupae from the Y-EGFP/bp + strain were irradiated with 30 Gy using a Co 60<br />
source, with subsequent appropriate crosses to select for Y-EGFP/bp + translocations for GSS.<br />
Results and Conclusions: Seven hundred (700) male progeny were screened, from which three<br />
potential lines were selected where all females had the mutant black pupae phenotype, while<br />
all males had the brown pupae (bp + ) wild type phenotype, in addition to green fluorescence<br />
observed under epifluorescent optics. These lines were maintained for four generations with no<br />
detection of recombinant individuals. One line had fertility higher than 50% and good fitness,<br />
which indicates good potential for development into a new GSS line allowing male-specific<br />
selection and marking necessary for SIT. This transgenic line is also being tested for remobilization<br />
of the vector to a new Y-linked autosomal site that may allow enhanced<br />
expression of the EGFP and sperm-specific DsRed markers. Y-linked fluorescence will provide<br />
the possibility for male-specific selection, especially if embryonic expression is detectable, for<br />
fluorescence-based embryonic sexing.<br />
Keywords: Mexfly, transgenic, genetic sexing strain, black pupae mutation, EGFP<br />
189
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
MITOCHONDRIAL DIVERSITY OF FIVE TUNISIAN CERATITIS CAPITATA<br />
POPULATIONS (DIPTERA: TEPHRITIDAE) EVALUATED BY SINGLE STRAND<br />
CONFORMATION POLYMORPHISM.<br />
M’saad Guerfali, Meriem 1* ; Hemdan, Heitham 1 ; Sellini, Lotfi 1 ; Raies, Aly 2 &<br />
Mathiopoulos, Kostas 3<br />
1 Unité de Production des mâles Stériles de la Cératite, Centre National des Sciences et Technologies Nucléaires,<br />
Technopole Sidi Thabet, 2020, Tunis, Tunisie. E-mail: msaad_tn@yahoo.fr.; 2 Laboratoire de Biomolécules<br />
Actives et de Microbiologie, Université Tunis El Manar Faculté des Sciences Naturelles de Tunis Campus<br />
Universitaire Tunis 2092 Tunisie; 3 Department of Biochemistry and Biotechnology, University of Thessaly,<br />
Ploutonos 26, Larissa 41221, Greece.<br />
Background: The Mediterranean fruit fly (Ceratitis capitata) (Diptera:Tephritidae)<br />
(Wiedemann) is an established pest in regions of north Africa. Little is known on the<br />
population structure and dynamics of north African populations. Nonetheless, effective control<br />
of this pest depends on knowledge of patterns of colonization of this economically important<br />
pest, such as the determination of invasion sources and the elucidation of demographic<br />
parameters. The aim of this present work was to analyze the genetic structure of medfly<br />
populations in this region, in an effort to understand the colonization process in the area.<br />
Methods: Mitochondrial DNA variation was studied in five natural populations from the<br />
northern, north-western, central and southern parts of Tunisia by amplifying three different<br />
mitochondrial loci (ND4, ND5 and COII) and subjecting them to Single Strand Conformation<br />
Polymorphism analysis (SSCP). The single stranded amplified DNA was electrophoresed on a<br />
non-denaturing polyacrylamide gel.<br />
Results: At least three distinct haplotypes were detected. These haplotypes are in the process of<br />
additional analysis: amplified bands are being isolated and sequenced in order to determine the<br />
exact nature of the detected polymorphisms. Further analysis will also resolve population<br />
structuring of Tunisian medfly populations.<br />
Conclusion: SSCP is a convenient, rapid and relatively inexpensive approach of surveying<br />
mitochondrial genetic variation. The three loci chosen for the analysis of medfly populations in<br />
Tunisia (ND4, ND5 and COII) present substantial sequence variation that can yield in<br />
meaningful results.<br />
Keywords : medfly, mitochondrial DNA, SSCP, polymorphism.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
TRANSGENIC CONDITIONAL LETHALITY STRAINS FOR THE BIOLOGICAL<br />
CONTROL OF ANASTREPHA SPECIES.<br />
Nirmala, Xavier; Schetelig, Marc F. & Handler, Alfred M.*<br />
Center for Medical, Agricultural, and Veterinary Entomology, USDA, ARS, 1700 SW 23 rd Drive, Gainesville,<br />
Florida, 32608 USA. Email: ahandler@ufl.edu<br />
Background: A variation of the sterile insect technique (SIT) for the biological control of pest<br />
species is the release of transgenic insects carrying dominant conditional lethal genes. Two<br />
approaches towards lethality include use of dominant temperature-sensitive (DTS) mutations,<br />
and tetracycline (tet)-suppressible lethality systems, in particular an embryo-specific system<br />
proven effective in medfly. For DTS lethality in Anastrepha spp. we tested a heat-sensitive<br />
mutation in the 20S proteasome subunit gene, Prosβ2 1 , first described in D. melanogaster. A<br />
missense mutation results in an antimorph at 29°C that disrupts proteasome function, resulting<br />
in late larval or pupal death. For tet-suppressible embryonic lethality we have initiated tests of<br />
genetic elements isolated from Anastrepha species for promoter-driver and lethal effector gene<br />
cassettes.<br />
Methods: For DTS lethality, proteasome subunit genes were isolated from A. suspensa<br />
embryonic cDNA libraries by degenerate PCR, with 5' and 3' genomic sequences isolated by<br />
inverse PCR. DTS mutant alleles, created by in vitro mutagenesis, were transformed into a<br />
wild strain using piggyBac transposon vectors. Homozygous lines for the DTS transgene were<br />
reared at 25°C and 29°C to evaluate viability from larval to adult life stages. For tetsuppressible<br />
lethality embryonic gene promoters and cell death gene cognates were identified<br />
from EST sequences by similarity searches and isolated by PCR.<br />
Results: The caribfly AsProsβ2 cognate was isolated and mutated to create a Gly170Arg<br />
substitution (AsProsβ2 1 ). Five transgenic lines homozygous for the mutant transgene pupated<br />
at frequencies similar to wild type at 25°C and 29°C, but exhibited pupal lethality rates of 96 to<br />
100% at 29°C. Embryonic gene cognates whose promoters may be used to drive lethality<br />
(linked to the tTA transcriptional activator), or from which dsRNA may act as lethal effectors<br />
(linked to the tet-response element; TRE) were isolated, including serendipity and slam. Their<br />
promoters were isolated, linked to tTA and are being tested with the TRE-hid ala5 lethal effector<br />
cassette (used in medfly) in A. suspensa transformant lines. Potential cell death gene cognates<br />
for hid, reaper and sickle were isolated from A. suspensa and A. ludens and are being tested<br />
functionally by cell death assays in Drosophila and Anastrepha embryonic cell lines.<br />
Conclusions: A DTS conditional lethal strain has been created for A. suspensa, yielding<br />
complete pupal mortality when reared at 29°C. While highly effective in caribfly, and likely<br />
useful in mexfly, post-larval lethality is less than optimal for immediate population control. A<br />
tet-suppressible embryonic lethality system, that uses lethal effector genes from Anastrepha,<br />
will potentially improve upon a similar system successfully tested in medfly. While embryonic<br />
lethality is highly preferable to pupal death, for transgenic release use of the two lethal systems<br />
together may be advantageous to ensure that reversion or modification of either one does not<br />
allow transgenics to survive in the field, that might result in re-population of the targeted<br />
species.<br />
Keywords: Anastrepha, SIT, conditional lethality, DTS mutations, tet-suppressible lethality<br />
191
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GENETICS ANALYSIS OF CERATITIS CAPITATA (WIEDEMANN) POPULATIONS<br />
USING MITOCHONDRIAL DNA SEQUENCES.<br />
Ochando, Maria Dolores; Hernandez, Sofia, Matallanas, Beatriz & Callejas, Carmen.<br />
Departamento de Genética, Facultad de CC. Biológicas, Universidad Complutense, C/ José Antonio Novais, nº2, 28040-<br />
Madrid, Spain. Email: dochando@bio.ucm.es.<br />
Background: The Ceratitis capitata species, is, among the Tephritidae family, one of the most<br />
damaging pests. From its putative origin in Africa, this species colonized the Mediterranean<br />
area and then, expanded all over the world with tropical or temperate climate. The present<br />
work reports a molecular phylogenetic study on geographic samples of Ceratitis capitata<br />
species.<br />
Methods: Flies from six different geographical origins, three from Spain, Mediterranean area,<br />
and other three from Africa origin (Kenya, Morocco, and Tunisia), were used for sequencing.<br />
A mitochondrial DNA region encompassing 344 bp have been sequenced. The segment include<br />
cytb, tRNA Ser and ND1 genes. DNA was extracted from individual flies, and amplified with<br />
primers CBF1: 5’-ACATGAATTGGAGCTCGACCAGT-3’ and N1R1: 5’-<br />
GGTACATTACCTCGGTTTCGTTATGAT-3’, designed by Dr. Fleming (Applied Plant<br />
Science, The Queen’s University of Belfast, United Kingdom) and based on Drosophila<br />
yakuba sequences (Clary and Wolstenholme, 1985). Total DNA was extracted from individual<br />
flies according to Reyes, Ochando et al. (1997). The polymerase chain reaction (PCR) was<br />
used to amplify and DNA amplifications were performed in 100 �l reaction volumes<br />
containing 4 �l DNA, 10 �l Ecotaq PCR buffer (Ecogen), 2mM MgCl2, 10 pmol CBF1, 10<br />
pmol NR1, 0.2 mM dNTPs and 2.5 U EcoTaq DNA polymerase (Ecogen). Double-stranded<br />
amplified products were purified using the High Pure PCR Product Purification Kit<br />
(Boehringer-Manheim) and used as templates for sequencing reactions. These reactions were<br />
performed using an ABI PRISM 377 DNA sequencer in the “Servicio de Secuenciación de la<br />
Universidad Complutense”.<br />
Results: The amplified segments correspond to positions 11523-11867 in the Drosophila<br />
yakuba mtDNA sequence. All the flies, European and African, showed the same haplotype.<br />
The obtained unique sequence was compared with those obtained in previous work and<br />
published in the GenBank (http://www.ncbi.nlm.nih.gov/Genbank) from C. cosyra, C. rosa, B.<br />
oleae, B. cucurbitae, B. dorsalis, and B. tryoni (accession numbers: AY096092, AY096094,<br />
AY096087, AY096081, AY096083, AY096088). Sequences were different for different<br />
species.<br />
Conclusions: The mitochondrial segment analyzed show a very high conservative sequence.<br />
However the time elapsed from colonization and the high density numbers of the populations<br />
of C. capitata, no variability is detected, and even, surprisingly enough, no variability is<br />
detected in African populations. The sequence is useful for taxonomic purposes.<br />
Keywords: Ceratitis capitata, mitochondrial sequences, haplotypes.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FEMALE-SPECIFIC EMBRYONIC LETHALITY SYSTEM FOR CERATITIS CAPITATA<br />
(WIEDEMANN).<br />
Ogaugwu, Christian E.* 1 ; Schetelig, Marc F. 1, 2 & Wimmer, Ernst A. 1<br />
1 Georg-August University Goettingen, Johann-Friedrich Blumenbach Institute of Zoology and Anthropology,<br />
Department of Developmental Biology, GZMB, Ernst-Caspari-Haus, Justus von Liebig-Weg 11, 37077<br />
Goettingen, Germany; Email: cogaugw@gwdg.de; 2 USDA/ARS, Center for Medical, Agricultural and Veterinary<br />
Entomology, Gainesville, FL 32608, USA.<br />
Background: The principle of the Sterile Insect Technique (SIT) involves mass production,<br />
mass rearing and mass release of sterile pest insects in the field to control the wild population.<br />
SIT is more effective when only sterile males are released and production of only males also<br />
reduces the general cost of carrying out an SIT programme. A female-specific lethality system<br />
based on a female specific splicing intron was developed for transgenic sexing in Ceratitis<br />
capitata, possibly to overcome the fitness problems of classical genetic sexing strains (GSS).<br />
However, this transgenic sexing system showed lethality predominantly during the pupa stage.<br />
To create an early-acting lethality system, we combined the principle of this female-specific<br />
lethality system with an embryonic lethality system to yield a female-specific embryonic<br />
lethality system.<br />
Methods: A sexing effector construct driven by the heterologous transactivator tTA and<br />
containing a female-specifically spliced intron inserted into the coding region of the<br />
proapoptotic gene hid was created and inserted into C. capitata via piggyBac-mediated<br />
transposition. Germ-line transformed flies were confirmed by fluorescence microscopy on G1<br />
adults. The resulting transgenic sexing effector lines were crossed with driver lines mediating<br />
early embryonic expression of the tetracycline-controlled tTA to generate lethal lines. To<br />
assess female-specific embryonic lethality, adults and larvae of these lethal lines were reared<br />
on tetracycline-free media and the sex of the subsequent adult progeny checked. Lethality was<br />
assessed for both heterozygous and homozygous adult progeny.<br />
Results: Different lethal lines showed varying strengths of female-specific embryonic lethality,<br />
with some lines showing complete female-specific lethality. This lethality does not affect male<br />
flies in any way observable to us. Female-specific lethality was suppressed on rearing adult<br />
flies and their larvae on 10µg/ml tetracycline media. In situ hybridizations showed the<br />
expected sequential activation of the components of this female-specific embryonic lethality<br />
system.<br />
Conclusion: The result of our study establishes female-specific embryonic lethality in the pest<br />
insect C. capitata, and such a system would also be relatively easy to develop for other<br />
tephritid species of agricultural importance. Early-acting female-specific lethality systems<br />
should offer cost-effective sexing of pest insect strains once introduced into SIT programmes.<br />
Keywords: Ceratitis capitata, integrated pest management, germ-line transformation, fluorescence microscopy,<br />
female-specific embryonic lethality<br />
193
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
MOLECULAR TOOLS FOR THE PATHWAY ANALYSIS OF CERATITIS CAPITATA.<br />
Ruiz-Arce, Raul* 1 ; Barr, Norman 1 , & De Meyer, Marc 2<br />
1 United States Department of Agriculture, Animal Plant Health Inspection Service, Center for Plant Health<br />
Science & Technology [USDA APHIS CPHST] Mission Lab, 22675 N. Moorefield Rd S6414, Edinburg, TX.<br />
78541 USA; Email: raul.a.ruiz@aphis.usda.gov; 2Royal Museum for Central Africa, Tervuren, Belgium.<br />
Background: Outbreaks of medfly, Ceratitis capitata (Diptera: Tephritidae), occur frequently<br />
in the United States. Determining the source of these introductions is key to the eradication of<br />
this pest. Unfortunately, morphological characters cannot be used to identify captures to<br />
populations-level. However, there are various molecular methods available for population<br />
analysis of medfly. These pathway tools are important to the eradication of this pest because<br />
they can be used to help develop better quarantine programs, for settling trade disputes, and to<br />
help provide valuable information for evaluating eradication programs. The USDA APHIS<br />
Mission Lab is assessing the utility of two mitochondrial and twenty-two nuclear markers for<br />
their use in the development of a comprehensive set of diagnostic tools for this pest. In order to<br />
facilitate the development, we are testing these markers with specimens from worldwide<br />
collections of recently-gathered wild and sterile insect technique (SIT) Mediterranean fruit<br />
flies.<br />
Methods: In order to evaluate the performance of nuclear and mitochondrial DNA for pathway<br />
analysis, we are examining the genetic diversity of numerous Mediterranean fruit fly<br />
collections. We are using the polymerase chain reaction and direct sequencing of mitochondrial<br />
COI and ND5/ND4 DNA. We are also exploring 22 previously developed microsatellite<br />
makers for their use in examining these collections.<br />
Results: Approximately 1900 individual flies from 53 collections and 19 different countries<br />
were gathered and submitted to the USDA APHIS Mission Lab fruit fly repository in the<br />
spring of 2010. Both COI and ND5/ND4 sequences from the flies were analyzed along with<br />
microsatellite loci. Our initial assessment of the mitochondrial DNA sequences suggests that<br />
haplotypes maintain a regional specificity. The microsatellite markers we evaluated show an<br />
increased resolution as compared to mtDNA and may serve for fine scale population analyses.<br />
Conclusions: Both mitochondrial and nuclear sets of markers provide have a high potential for<br />
effective pathway analysis tool development. Further work is needed in order to properly<br />
evaluate their utility and develop an effective set of reliable pathway analysis tools.<br />
Keywords: Mediterranean fruit fly, PCR, DNA
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
PRELIMINARY STUDIES ON THE PRESENCE OF TWO HAPLOTYPES OF THE OLIVE<br />
FLY SYMBIONT “CANDIDATUS ERWINIA DACICOLA” IN ITALY.<br />
Savio, Claudia*; Martinez-Sañudo, Isabel; Mazzon, Luca & Girolami, Vincenzo.<br />
Dipartimento di Agronomia Ambientale e Produzioni Vegetali (DAAPV), Università di Padova. AGRIPOLIS –<br />
Viale dell’Università 16 – 35020 Legnaro, Padova (Italy). Email: claudia.savio@unipd.it<br />
Background: A very close association between the olive fly Bactrocera oleae (Rossi) (Diptera:<br />
Tephritidae) and bacteria has been known for more than a century since Petri firstly described<br />
Pseudomonas savastanoi as its symbiont. Recently, using molecular techniques, the presence<br />
of an hereditary, host specific, unculturable symbiotic bacterium, designated as ‘Candidatus<br />
Erwinia dacicola’, has been confirmed in all wild specimens even when adults emerged from<br />
previously surface sterilized pupae. The present work aims to characterize the possible<br />
existence of different haplotypes of “Ca. E. dacicola” and their distribution in Italian territory.<br />
Methods: Two-three day-old flies (n= 281), collected from 26 locations across the Italian range<br />
of the olive tree, were aseptically dissected to remove the oesophageal bulbs. The DNA of their<br />
content was extracted using a salting-out protocol, and an amplification by PCR was performed<br />
on the 16s rRNA, using universal bacterial primers. The contents of some olive fly oesophageal<br />
bulbs were also cloned using the P-GEM-T Easy vectors (Promega). PCR products and clones,<br />
randomly chosen from the constructed libraries, were subjected to DNA sequencing and<br />
restriction enzyme detection.<br />
Results: With the exception of a few cases (1.15%) GenBank similarity search showed that the<br />
amplified DNA fragment was similar to 16S ribosomal RNA gene of “Ca. E. dacicola”. When<br />
aligning the sequences obtained two “Ca. E. dacicola” haplotypes were found, one being<br />
slightly more prevalent than the other (56%). Populations were significantly different from<br />
each other in the relative frequencies of the two haplotypes; in particular there seems to be a<br />
positive, significant correlation in their distribution along the Tyrrhenian coast that does not<br />
occur along the Adriatic coast. Moreover the two haplotypes never co-exist in the same fly, as<br />
confirmed by cloning. Populations monitored for three consecutive years show no significant<br />
differences in the haplotype frequency.<br />
Conclusions: “Ca E. dacicola” presents at least two haplotypes in Italy whose distribution is<br />
not random but significantly different in different regions with an evident gradient of transition,<br />
not influenced by sampling in terms of time. Extending the sampling to the Mediterranean area<br />
will be the next step in this research.<br />
Keywords: Bactrocera oleae, “Candidatus Erwinia dacicola”, symbiont, haplotypes.<br />
195
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
OLFACTION IN INSECTS: MOLECULAR CHARACTERIZATION AND EXPRESSION<br />
PROFILES OF PUTATIVE PHEROMONE-BINDING PROTEIN GENES FROM THE<br />
MODEL PEST SPECIES CERATITIS CAPITATA (DIPTERA:TEPHRITIDAE).<br />
Siciliano, Paolo 1 ; Scolari, Francesca 1 ; Gabrieli, Paolo 1 ; Gomulski, Ludvik M. 1 ; Guglielmino, Carmela R. 2 ;<br />
Falchetto, Marco 1 ; Bonomi, Angelica 1 ; Malacrida, Anna R. 1* & Gasperi, Giuliano 1<br />
Dipartimento di Biologia Animale, Università di Pavia, Via Ferrata 1, 27100 Pavia, Italy. 2 Dipartimento di<br />
Genetica e Microbiologia, Università di Pavia, Via Ferrata 1, 27100 Pavia, Italy.<br />
Background: In insects, pheromones play an important role in intra- and inter-sex<br />
communication, inducing specific behavioural responses in terms of sexual attraction for matefinding,<br />
mating aggregation, or host-marking of oviposition sites. Odour perception is<br />
regulated by a fine molecular pathway that involves multigene families including odorantbinding<br />
proteins (OBPs) and chemosensory proteins (CSPs). According to the type of ligand,<br />
OBPs are classified in general odorant-binding proteins (GOBPs) and pheromone-binding<br />
proteins (PBPs). Although notable progress in the study of pheromone perception has been<br />
achieved in several species, the molecular basis underlying the outstanding selectivity and<br />
sensitivity of olfaction in pests such as the Mediterranean fruitfly, Ceratitis capitata, is still<br />
unknown. In this study we aimed at furnishing the first insights on the pheromone perception<br />
process in this species.<br />
Methods: A well established C. capitata ISPRA strain reared under standard conditions was<br />
used for the experiments. Two different cDNA libraries were investigated by BlastX and<br />
tBlastX analyses to identify medfly PBP homologues. The five sequences that showed high<br />
similarity to PBP genes of other insect species were first characterized (cloning, 5’-3’RACE-<br />
PCR, PCR) and then analyzed to define their expression profiles in relation to sex, tissue,<br />
maturation and mating status (RT-PCR, Northern Blot, qPCR). In silico analyses were<br />
performed to verify the presence of all the features of the PBP family and to obtain a<br />
hypothetical 3D protein structure.<br />
Results: We report the cloning, characterization and expression profiles of five putative PBP<br />
genes (CcPBP1 to CcPBP5) isolated from two cDNA libraries. Molecular analyses revealed<br />
the presence of all the features of the PBP family (signal peptide, hydrophobic domain, six<br />
conserved cysteines). Gene structures showed high similarity to Drosophila putative<br />
homologues, except for the presence of longer introns in the medfly. Expression profile<br />
analyses showed that i) all the genes were highly over-expressed in head; ii) transcription<br />
levels were higher in females than in males, apart from CcPBP4; iii) sexually mature males and<br />
females showed higher transcript levels of each gene; iv) after mating the transcript levels in<br />
females tended to decrease, while males did not experience such a strong effect.<br />
Conclusions: The finding of multiple potential PBPs in the medfly suggests that PBPs may also<br />
play a role in the differentiation of pheromone components by differential binding capacities.<br />
Future biochemical, functional, behavioural and transgenic assays, will clarify the mechanisms<br />
underlying odour and pheromone perception. This knowledge will enable the identification of<br />
PBP ligands, to determine their roles in medfly chemoreception and, in the long term, to use<br />
pheromone components as target molecules in field studies to develop synthetic attractants or<br />
repellents for pest control programs.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EXPRESSED SEQUENCE TAGS (ESTS) IN THE OLIVE FLY, BACTROCERA OLEAE<br />
(DIPTERA: TEPHRITIDAE).<br />
Tsoumani, Konstantina 1 ; Augustinos, Antonios 1,2 ; Kakani, Evdoxia 1 ; Drosopoulou,<br />
Eleni 3 ; Mavragani-Tsipidou, Penelope 3 & Mathiopoulos, Kostas 1<br />
1 Department of Biochemistry and Biotechnology, University of Thessaly, Larissa, Greece; 2 Department of<br />
Biology, University of Patras, Rion, Greece; 3 Department of Biology, Aristotle University of Thessaloniki,<br />
Thessaloniki, Greece<br />
Background: The olive fruit fly, Bactrocera oleae, is the major pest of the olive tree. Despite<br />
its importance, very little genetic and molecular knowledge is available. The present study is a<br />
first attempt to identify and partially characterize B. oleae expressed sequence tags (ESTs). A<br />
few possible uses are also explored.<br />
Methods: Random cDNA clones were isolated from an adult B. oleae library and the generated<br />
ESTs were annotated through BLASTX similarity searches. A few of them were physically<br />
mapped by in situ hybridization. Codon usage was determined for the resulting open reading<br />
frames. Seventeen Exon-Primed-Intron-Crossing (EPIC) primers were developed and tested on<br />
eleven Tephritids.<br />
Results: One hundred and ninety-five randomly selected cDNA clones and the generated ESTs<br />
were annotated through BLASTX similarity searches. A set of 159 unique putative transcripts<br />
were functionally assigned using Gene Ontology terms in broad categories of biological<br />
process, molecular function and cellular component based on D. melanogaster matches.<br />
Moreover, the cytogenetic location of 35 ESTs by in situ hybridization to B. oleae polytene<br />
chromosomes was defined. The deduced codon usage of the isolated ESTs suggested a<br />
conserved pattern of B. oleae with its closest relatives. Additionally, the comparative analysis<br />
of B. oleae ESTs with the homologous D. melanogaster genes led to the development of 17<br />
nuclear EPIC-PCR markers for the amplification of intron sequences of 11 Tephritids.<br />
Sequencing analysis of several cross-amplified intron sequences revealed a high degree of<br />
conservation among Bactrocera species and a varying transferability of the generated markers<br />
across the examined genera.<br />
Conclusions: The generated B. oleae EST collection constitutes the largest set of putative<br />
expressed genes in the species. The resulting low resolution EST map more than doubles the<br />
available entry points to the insect’s genome and can assist syntenic comparisons with other<br />
distant species. EPIC analysis can provide a useful tool for the clarification of phylogenetic<br />
relationships among different species, particularly in cases of species complexes.<br />
Keywords: Gene Ontology (GO), in situ hybridization, EPIC markers, codon usage<br />
197
Session 4<br />
Risk Assessment, Quarantine &<br />
Post-harvest Treatments
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DETECTION OF ANASTREPHA GRANDIS (MACQUART) IN CUCURBITS WITH THE<br />
USE OF INTEGRATED MEASURES IN A SYSTEM APPROACH FOR PEST RISK<br />
MANAGEMENT IN THE STATE OF GOIÁS, BRAZIL.<br />
Caixeta, Claudini V. D. 1* ; Pereira, André F. 2 ; Veloso, Valquíria da R. S. 3 ; Rabelo, Lílian<br />
R. S. 4 & Rios, Anderli D. F. 3<br />
1 Superintendência Federal de Agricultura em Goiás/MAPA/DSV, Praça Cívica n° 100, 6° andar, Centro, Goiânia-<br />
Goiás, Brasil, claudini.caixeta@agricultura.gov.br. 2 Instituto Federal de Educação Ciência e Tecnologia de<br />
Brasília, Campus Planaltina, Rodovia DF 128, Km 21, Zona Rural - Km 18 da BR 020, Caixa Postal: 08202,<br />
Planaltina-DF, Brasil; Universidade Federal de Goiás (UFG). 3 Escola de Agronomia e Engenharia de Alimentos,<br />
Campos Samambaia, Caixa Postal 131; Goiânia-GO, Brasil; 4 Agência Goiana de Defesa Agropecuária-<br />
AGRODEFESA, Av. Portugal, 591, qd.j-10, Lt.06, Setor Oeste, Goiânia-GO.<br />
Background: The presence of fruits flies in trade fruit crops, represents an obstacle to the<br />
production and marketing of fresh fruits in Brazil as well as worldwide, due to the direct<br />
damages caused by the attack of the larvae as well as to the impediments to exportation, in the<br />
form of quarantine measures imposed by importer countries. Fruits of the Cucurbitaceae family<br />
are hosts of Anastrepha grandis (Macquart), which is among the fruit fly species that are<br />
subject to quarantine restrictions. Detection survey of Anastrepha grandis was concluded to<br />
provide information regarding the occurrence of this species. The results might serve as<br />
scientific basis so that several municipalities in Goiás State can implement the Integrated<br />
Measures in a Systems Approach for Pest Risk Management and ensure the phytosanitary<br />
security of Brazil’s fruit exportation.<br />
Methods: The monitoring was conducted at productions units in the properties registered in the<br />
Integrated Measures in a Systems Approach for Pest Risk Management in the municipalities of<br />
Carmo do Rio Verde, Cristalina, Goianésia, Ipameri, Itapuranga, Jaraguá, Maurilândia, Rio<br />
Verde, Santa. Helena, São Miguel do Araguaia and Uruana, on watermelon, melon and<br />
pumpkin crops. It used McPhail traps with 5% hydrolyzed corn protein (food attractant), with<br />
density of 1 trap/5 hectares and inspection on a weekly basis, within the period of December<br />
2004 to February 2010. After each service, the traps were washed and the attractant was<br />
replaced. All the captured insects were transferred to vials containing 70% alcohol, and sent to<br />
the Insect Identification Laboratory at the Escola de Agronomia e Engenharia de Alimentos da<br />
Universidade Federal de Goiás (UFG) for further identification.<br />
Results: The Index fly/trap/day keep on the acceptable limit in the Brazilian legislation for<br />
implementation and maintenance the Systems Approach for Pest Risk Management. Number<br />
of specimens of Anastrepha. grandis (Macquart) detected in 2008 and 2009: Ipameri (01),<br />
Cristalina (01), Jaraguá (01), Uruana (03), Rio Verde (01) and Goianésia (07).<br />
Conclusions: The species Anastrepha grandis (Macquart) was detected in 2008 and 2009 in the<br />
municipalities of Ipameri, Cristalina, Jaraguá, Uruana, Rio Verde and Goianésia, at Goiás<br />
State, in Brazil. The results show the low density of A. grandis (Macquart) registered in the<br />
fields of production of cucurbits throughout the year and in the general population remained<br />
within non-detectable levels in the most of the years. Detection survey confirm previous<br />
studies which concluded that the pest populations have low density, therefore, they are more<br />
important due to quarantine controls, than the damages it actually causes.<br />
Keywords: Cucurbitaceae, fruit fly, Anastrepha grandis, Pest Risk Management, detection.<br />
201
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IMPLANTATION OF THE INTEGRATED MEASURES IN A SYSTEMS APPROACH FOR<br />
PEST RISK MANAGEMENT IN THE STATE OF GOIÁS, BRAZIL.<br />
Caixeta, Claudini V. D. 1 ; Pereira, André F. 2 ; Veloso, Valquíria da R. S. 3 ; Rabelo, Lílian<br />
R. S. 4 & Rios, Anderli D. F. 3<br />
1 Superintendência Federal de Agricultura em Goiás/MAPA/DSV, Praça Cívica n° 100, Centro, Goiânia-Goiás, Brasil,<br />
claudini.caixeta@agricultura.gov.br; 2 Instituto Federal de Educação Ciência e Tecnologia de Brasília, Campus Planaltina,<br />
Rodovia DF 128, Km 21, Zona Rural - Km 18 da BR 020, Caixa Postal: 08202, Planaltina-DF, Brasil; Universidade Federal de<br />
Goiás (UFG); 3 Escola de Agronomia e Engenharia de Alimentos, Campos Samambaia, Caixa Postal 131; Goiânia-GO, Brasil;<br />
4 Agência Goiana de Defesa Agropecuária-AGRODEFESA, Av. Portugal, 591, qd.j-10, Lt.06, Setor Oeste, Goiânia-GO.<br />
Background: The presence of fruit flies in trade fruit crops represents an obstacle to the<br />
production and marketing of fresh fruits in Brazil as well as worldwide, due to the direct<br />
damages caused by the attack of the larvae as well as to the impediments to exportation, in the<br />
form of quarantine measures imposed by importing countries. Fruits of the Cucurbitaceae<br />
family are hosts of Anastrepha grandis (Macquart), which is among the fruit fly species that<br />
are subject to quarantine restrictions. To export fresh fruits of cucurbits crops to countries<br />
devoid of fruit flies, the countries with the presence of fruit flies should have their fruit coming<br />
from pest free areas or areas with implementation of the Integrated Management Measures for<br />
Risk Management (SMR) officially recognized, ensuring products free of fruit flies. The work<br />
aimed at implementing the Integrated Measures in a Systems Approach for Pest Risk<br />
Management - RMS for pest Anastrepha grandis (Macquart) in the State of Goiás, in order to<br />
meet the requirements of importing countries which impose fitossanitary restrictions of<br />
cucurbitaceous trade.<br />
Methods: The implantation of the RMS is subject to the detection of the pest Anastrepha<br />
grandis (Macquart) in the proposal area, according to Instruction Normative 16/2006, which<br />
establishes the monitoring for at least six uninterrupted months before the request for official<br />
recognition. The Farms of properties inscribed on watermelon, melon and pumpkin cultivation,<br />
were registered in the State Board of Plant Protection Plant and monitoring Anastrepha grandis<br />
(Macuart) within the period of December 2004 up to June 2005, from April to September 2007<br />
and from April to September 2008. The farms were audited and recognized by the Plant<br />
Protection Division of Ministry of Agriculture, Livestock and Food Supply through the<br />
Normative Instruction 41/2006, 22/2008 and Resolution 1/ 2009.<br />
Results: The state of Goiás in Brazil, implemented and obtained official recognition by the<br />
Ministry of Agriculture, Livestock and Food Supply the Mitigation Risk System -RMS for pest<br />
Anastrepha grandis (Macquart) in 2006, 2008 and 2009 in the municipalities of Carmo do Rio<br />
Verde, Cristalina, Ipameri, Itapuranga, Jaraguá, Maurilândia, Rio Verde, Santa Helena and<br />
Uruana.<br />
Conclusions: The implantation of SMR has allowed, since 2006, the exportation of<br />
watermelon, melon, pumpkin fresh fruits which led to the expansion of trade and the opening<br />
of new markets for the State of Goiás.<br />
Keywords: Cucurbitaceae, fruit flies, Integrated Measures in a Systems Approach for Pest Risk Management,<br />
Anastrepha grandis, exportation
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
WHEN FRUIT FLIES FLY. DATA ON THREE YEARS OF TEPHRITIDAE<br />
INTERCEPTION BY THE FRENCH NPPO IN ROISSY CHARLES-DE-GAULLE<br />
AIRPORT.<br />
Balmès, Valérie 1 , Germain, Jean-François1*; Salin, Daniel 2 & Gourdon, Aude 2<br />
1 Laboratoire National de la Protection des Végétaux, station de Montpellier. CBGP Campus international de<br />
Baillarguet CS 30016, FR-34988 Montferrier-sur-Lez cedex; E-mail: germain@supagro.inra.fr; 2 SRAL-DRAAF<br />
antenne régionale de Roissy Charles de Gaulle BP 10112 FR-95701 Roissy CDH cedex.<br />
Background: All non european Tephritidae are in quarantine pest list for EU area. In Roissy<br />
Charles de Gaulle airport, the NPPO inspectors control commodities (fresh fruits/ vegetables)<br />
that come from different parts of the world. Main pathways, commodities and fruit flies groups<br />
are presented.<br />
Methods: The inspectors are formed to recognized Tephritidae larva and other quarantine pests.<br />
When they found larvae, several are send to LNPV* for confirmation allowing official<br />
notification. If possible, the other available larvae are reared and resulting adults are sent to<br />
LNPV for species identification. Presented datas correspond to the results of these rearings<br />
during the years 2007 to 2009.<br />
Results: Over the followed period 716 rearings gave adults identified to species (5433<br />
specimens). Four genus are represented, by order of importance: Bactrocera, Ceratitis,<br />
Anastrepha and Dacus. Fruits Flies were taken on fourteen varieties of fresh fruits/vegetables.<br />
The main is Mangifera indica, followed by Annona spp., Capsicum spp., Syzygium spp. and<br />
Psidium guajava. The geographical origin is mainly Asia, then Africa and to a lesser extent<br />
neotropical area.<br />
Conclusion: Data collecting during last there years allow identifying possible pathways of<br />
introduction of the fruit flies in EU. Only species of agronomic importance are recorded<br />
pointing out a strong link between the species and its host-plant (ex: Bactrocera invadens and<br />
mango) or between the species and the area of origin (ex: Anastrepha spp. and neotropical<br />
area). The observation of Bactrocera invadens in the records shows the importance of the<br />
survey and the interest of the identification at species level in following pathways.<br />
Keys words: quarantine, pathway, interception<br />
203
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FRUIT FLY MANAGEMENT BEGINS AT HOME.<br />
Barnes, Brian* 1 & Sutherland, Ian 2<br />
1 ARC Infruitec-Nietvoorbij Institute for Fruit, Vine & Wine, Stellenbosch, South Africa; Email:<br />
barnesb@arc.agric.za; 2 SIT Africa (Pty) Ltd, Stellenbosch, South Africa.<br />
Bacground: Three species of fruit flies of economic importance occur in South Africa -<br />
Mediterranean fruit fly (Ceratitis capitata), Natal fruit fly (C. rosa) and marula fruit fly (C.<br />
cosyra). Their pest status is exacerbated by the fact that they are international quarantine pests,<br />
restricting the free trade of export fruit. Chemical options for fruit fly control are limited to<br />
baits comprising an attractant and an insecticide, and the use of "attract and kill" technology<br />
(bait stations). The sterile insect technique is also used in certain areas against C. capitata.<br />
None of these strategies is regarded as a 'stand-alone' - all possible strategies should be<br />
incorporated in an integrated management approach also involving vineyard/orchard sanitation<br />
and host plant management. Success with fruit fly control in South Africa varies; poor control<br />
is mostly due to non-compliance with recommendations, especially sanitation and host plant<br />
management. Understanding fruit fly behaviour, and in particular their migration behaviour<br />
and easy adaptation to home garden environments, can play a major role in limiting infestation<br />
in commercial plantings. In this regard a trial was carried out to monitor the occurrence of C.<br />
capitata and C. rosa simultaneously in a block of commercial table grape vineyards and in<br />
nearby home gardens over two seasons, to identify the time and place during the season when<br />
fruit fly populations are most vulnerable to management interventions, and to identify periods<br />
in the season and target areas for optimal fruit fly control.<br />
Materials and Methods: A vineyard of five adjacent blocks of export table grape (±12 ha) with<br />
reportedly high populations of fruit flies were monitored from December 2006 to October 2008<br />
on a farm in the Worcester district of the Western Cape. Two home gardens in a residential<br />
area approximately 400 m from the vineyard site were simultaneously monitored. Five<br />
Chempac® traps baited with 3-component lure were placed in the table grape vineyard, and<br />
one each in suitable shade trees in the two home gardens, and serviced monthly. All male and<br />
female fruit flies were recorded. The vineyards were harvested from January to April.<br />
Results: In the vineyard blocks, very high fruit fly populations occurred from March to June,<br />
while fruit flies were absent or in very low numbers from July to February. In the residential<br />
gardens, moderate but fluctuating populations occurred throughout the year, including July to<br />
February when fruit flies were absent in the vineyards.<br />
Conclusions: The results highlight the role urban areas play in fruit fly infestation levels in<br />
commercial fruit. Adults leave commercial fruit plantings in early winter, seeking shelter, food<br />
and breeding sites in residential gardens with host plants year-round. They re-infest<br />
commercial vineyards and orchards in summer when ripe fruit is available and susceptible. The<br />
greatest control effort should therefore be made in all gardens from early winter to early<br />
summer to limit the size of populations that will later migrate to vineyards and orchards in<br />
summer. All possible strategies should be used: fruit fly bait, bait stations, host plant<br />
management, sanitation, and SIT where applicable.<br />
Keywords: Monitoring, migration, commercial fruit, home gardens, management strategy
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
THE POTENTIAL GEOGRAPHIC DISTRIBUTION OF BACTROCERA ZONATA<br />
(SAUNDERS) IN EUROPE AND THE MEDITERRANEAN BASIN.<br />
Cobos-Suárez, José M. 1 ; Catalán-Ruescas, Diana 2 & Guitián-Castrillón, José M. 2* .<br />
1 Ministerio de Medio Ambiente y Medio Rural y Marino (MARM), Subdirección General de Sanidad de la<br />
Producción Primaria, C/ Alfonso XII 62; 28014 Madrid (Spain). Email: jcobossu@marm.es; 2 Tecnologías y<br />
Servicios Agrarios, S.A. (Tragsatec) C/ Hnos. García Noblejas 37c, 2 nd floor; 28037 Madrid (Spain).<br />
Background: Ten years ago, the peach fruit fly (PFF) Bactrocera zonata (Saunders) seemed to<br />
present a lesser risk for Europe than other fruit flies as its geographic distribution was restricted<br />
to tropical areas. However its rapid spread and establishment in Egypt has confirmed that the<br />
PFF should be treated as a priority, at least, by Mediterranean countries. In this article we have<br />
developed a predicted distribution of the PFF for Europe and the Mediterranean Basin using<br />
CLIMEX modelling package.<br />
Methods: Climate matching software CLIMEX was used to synthesize available information<br />
from diverse sources and assess the invasion potential of the PFF within Europe and the<br />
Mediterranean Basin. A Composite Match Index (CMI) was determined with the 'Match<br />
Climates' function in order to match: (a) the ranges in Egypt where the fly is introduced with<br />
its native range in India and Pakistan; (b) the ranges in Europe and the Mediterranean Basin<br />
with the current distribution of the PFF. The Ecoclimatic Index (EI) was estimated in order to<br />
have an overall measure of favourableness of each European location for permanent occupation<br />
by the PFF.<br />
Results: Broadly speaking, the climate of India is not similar to that of Egypt. On the other<br />
hand, the climate of Pakistan shows significant similarities with most of the Egyptian locations.<br />
The climatic similarity between Egyptian and the European locations is variable: Alexandria<br />
and El Tur, where the PFF is known to occur, are similar to several locations of Spain, Greece,<br />
Italy and Portugal but the climate of El Cairo is not comparable to any location in Europe.<br />
Regarding the EI, more than 40 locations within Europe have obtained an EI > 30, which<br />
means that their ecoclimatic conditions are very favourable for the pest establishment. In these<br />
places the PFF is predicted to have an economic impact.<br />
Conclusions: The locations within the areas of origin of the PFF are not climatically similar.<br />
This may lead to the contention that the Indian populations of the PFF have different climatic<br />
requirements to the populations found in Pakistan or those introduced in Egypt and the Near<br />
East. The climate model supports that Mediterranean and semi-arid climates are predicted to be<br />
suitable for the long-term survival of the PFF. Due to the predicted capacity of PFF to<br />
withstand drier climatic conditions than those traditionally associated to this fly, much of the<br />
Mediterranean Basin could be at risk of invasion.<br />
Keywords: Climate matching, Pest risk prediction, CLIMEX, invasion potential, climatic conditions.<br />
205
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DETERMINATION OF THE MOMENT UNDER WHICH VERNA LEMON (CITRUS<br />
LEMON) IS SUSCEPTIBLE TO CERATITIS CAPITATA IN THE SPANISH SOUTHEAST.<br />
Quinto, Vicente* 1 ; Fernández, Milagros 1 ; Arques, Eva 1 ; Lucas, Alfonso 2 & Lacasa,<br />
Alfredo 3<br />
1 Empresa de Transformación Agraria, S.A. (Tragsa) - Servicio de Sanidad Vegetal de la Consejería de Agricultura<br />
y Agua de la Comunidad Autónoma de la Región de Murcia. C/ Mayor, s/n, 30150 La Alberca (Murcia), España.<br />
E-mail: vicente.quinto@hotmail.com; 2 Servicio de Sanidad Vegetal de la Consejería de Agricultura y Agua de la<br />
Comunidad Autónoma de la Región de Murcia. C/ Mayor, s/n, 30150 La Alberca (Murcia), España.<br />
3 Biotecnología y Protección de Cultivos, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario.<br />
C/ Mayor, s/n, 30150 La Alberca (Murcia), España.<br />
Background: Until 2009, the lemon was not included in the list of quarantine host of Ceratitis<br />
capitata, although there was evidence of isolated incidents of infested fruits. Therefore special<br />
regulatory rules for international trade were not established. In 2006, at the end of the export<br />
campaign to US, fruits were found infested on arrival from a batch of “Verna” lemons from the<br />
Spanish mainland southeast. The singular climatic and agronomic conditions of the 2006<br />
campaign brought about harvesting of fruits damaged or senescent which were infested. In<br />
order to adjust the conditions of harvesting of fruits for export, it was needed to determine the<br />
susceptibility time frame and the conditions which allows the complete development of C.<br />
capitata.<br />
Methods: Over the last two campaigns Verna lemons have been regularly sampled from three<br />
orchards registered in the official record for the export of fruits to US. Additionally, C. capitata<br />
monitoring was established for the three orchards along the study period. Fruit sampling started<br />
at fruit colour change until early July.Commercial size fruits were taken each week during the<br />
first month and every 3 weeks since the fruits were completely yellow. For each orchards and<br />
group of 100 fruits it was determined: a) Natural infestation, keeping them in incubation for 90<br />
days at 25ºC, between 70 and 100% relative humidity and a photoperiod of 14:10 hours<br />
light:dark. b) Fruit susceptibility by artificial infestation (by exposing the collected fruits to a<br />
population of C. capitata, fruits were incubated under the same conditions).<br />
Results: Natural infestation in the orchard occurs at a time when the levels of capture of adults<br />
in the plots are highest. Shortly before detection of natural infestation, a small proportion of<br />
fruits were susceptible to artificial infestation. In controlled conditions C. capitata was able to<br />
infest and complete development in those yellow Verna lemons.<br />
Conclusions: The results show that natural infestation of Verna lemons depends on the<br />
characteristics of the fruits (colour and damage) and on the adult immigrants populations that<br />
arrive to the plots from other host plants.<br />
Keywords: Ceratitis capitata, quarantine, susceptibility.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
INDICATOR PARAMETERS OF SUSCEPTIBILITY OF VERNA LEMONS (CITRUS<br />
LEMON) TO CERATITIS CAPITATA (WIEDEMANN).<br />
Quinto, Vicente* 1 ; Fernández, Milagros 1 ; Arques, Eva 1 ; Lucas, Alfonso 2 & Lacasa,<br />
Alfredo 3<br />
1 Empresa de Transformación Agraria, S.A. (Tragsa) - Servicio de Sanidad Vegetal de la Consejería de Agricultura<br />
y Agua de la Comunidad Autónoma de la Región de Murcia. C/ Mayor, s/n, 30150 La Alberca (Murcia), España.<br />
E-mail: vicente.quinto@hotmail.com; 2 Servicio de Sanidad Vegetal de la Consejería de Agricultura y Agua de la<br />
Comunidad Autónoma de la Región de Murcia. C/ Mayor, s/n, 30150 La Alberca (Murcia), España.<br />
3 Biotecnología y Protección de Cultivos, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario.<br />
C/ Mayor, s/n, 30150 La Alberca (Murcia), España.<br />
Background: Is abundant the bibliography which report contamination episodes by Ceratitis<br />
capitata of certain varieties of lemons when these are overripe. In the summer of 2006 adults<br />
of this species were found in an export batch of “Verna” lemon from Spanish southeast to US.<br />
Following this event were initiated studies to try to determine some characteristic of the fruit,<br />
to distinguish the susceptible fruits that can be infested of non-susceptible, in order to facilitate<br />
regulatory inspections in the marketing.<br />
Methods: Over the last two campaigns have been harvested “Verna” lemon fruits in orchards of<br />
the three most important production areas in the Region of Murcia, from the months of<br />
November to June-July. The samples were collected each week during the colour change and<br />
every 3 weeks later. Some of the fruits were used to determine the natural infestation and to<br />
evaluate the susceptibility from exposure to adult populations of Ceratitis capitata in artificial<br />
conditions. Another part was used to determine the following parameters: acidity of the juice,<br />
ºBrix, colour, peel thickness, equatorial diameter, hardness of the peel, chemical composition<br />
of the peel (albedo and flavedo), etc. The correlations between these parameters have been<br />
studied and the infestation in the different sampling dates.<br />
Results: Not has been found a representative parameter of the characteristics of the fruit that is<br />
associated with the susceptibility or resistance to infestation by Ceratitis. The parameter more<br />
indicative of the susceptibility is the ratio between the force required to break the peel and the<br />
distance at which rupture occurs. Also, the composition of the albedo would be indicative of<br />
the susceptibility or resistance, because the larvae did not complete their postembryonic<br />
development, when feed on albedo<br />
Conclusions: Any parameter representative of the fruit that is closely related to the<br />
susceptibility or resistance of fruit infestation by Ceratitis has been found. We need to study<br />
more thoroughly the ratio between the force required to break the peel and the distance at<br />
which rupture occurs, as well as the chemical composition of albedo.<br />
Keywords: fruit fly, export protocol, susceptibility.<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CONTRIBUTION FOR DEVELOPMENT OF A CERATITIS CAPITATA (WIEDEMANN)<br />
RISK MODEL IN TERCEIRA ISLAND.<br />
Lopes, D.J.H. *1 ; Pimentel, R. 1 ; Macedo, N. 1 ; Zorman, M. 3 ; Ventura, L.B. 1; Dantas,<br />
Luis 2 ; Mexia, A.M.M. 4 ; & Mumford, John M. 5<br />
1 Universidade dos Açores, Centro de Biotecnologia dos Açores, Departamento de Ciências Agrárias, Secção de<br />
Protecção de Plantas, 9701-851 Terra chã. E-mail: dlopes@notes.angra.uac.pt; 2 Programa Madeira-Med, Estrada<br />
Eng. Abel Vieira, 262, 9135-260 Camacha, Madeira, Portugal. 3 Faculty of Agriculture, Vrbanska 30 , SI-2000<br />
Maribor, Slovenia. 4 Universidade Técnica de Lisboa, Instituto Superior de Agronomia, Departamento de<br />
Protecção de Plantas e Fitoecologia, Tapada da Ajuda. 5 Imperial College of Science, Technology and Medicine,<br />
Department of Environmental Science and Technology, Silwood Park, Ascot, United Kingdom.<br />
Background: The Medfly (Ceratitis capitata Wied.) (Díptera: Tephritidae) worldwide<br />
biological adaptability as well as the careless fruit transportation, sometimes infested with eggs<br />
or even larves, were two the most important factors which allowed the fast dispersion of this<br />
pest over many continental areas and islands. The behavioral study, as well as the fight and<br />
prevention studies about this insect has became very important, because there has been a bigger<br />
occurrence of this pest over several fruit cultures, carrying sometimes severe economic impacts<br />
to the fruit producers. In the Azores Islands, especially on Terceira Island the behavioral study<br />
of this insect has been done by Interfruta and Interfruta II Investigation Projects. Their studies<br />
have been covering the population dynamics, dispersion, fruit infestation and some control<br />
measures. This work will try to expand the horizons of these studies with the creation of a<br />
model about risk prediction of C. capitata for Terceira Island.<br />
Methods: For model development, traps capture records from some works done in Interfruta II<br />
project are going to be used to try to establish a reasonable multi-correlation ratio between<br />
these records and variables such as climatic, soil occupation and fruit availability.<br />
Keywords: Medfly, Risk model, Interfruta projects
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
THE BANANA FLY, BACTROCERA MUSAE (TRYON): IS IT A BANANA PEST?<br />
Mararuai, Amanda 1,2 & Clarke, Anthony R. 1,3*<br />
1Queensland University of Technology (QUT), Discipline of Biogeosciences, Faculty of Science & Technology,<br />
Brisbane, Queensland 4001, Australia; 2 [PNG] National Agricultural Research Institute Dry Lowlands Program<br />
(NARI DLP), Port Moresby, Papua New Guinea; 3 Cooperative Research Centre for National Plant Biosecurity<br />
(CRCNPB), Canberra, Australia; Email: a.clarke@qut.edu.au.<br />
Background: The Pest Risk Analysis (PRA) is a central plank of international phytosanitary<br />
regulations. A fundamental aspect of PRAs is knowing the risk posed by different pests.<br />
Unfortunately, for many crops susceptible to fruit flies, PRAs are often based on generic fruit<br />
fly host lists, in which fruit varietal differences, ripening stages and field usage of hosts is<br />
rarely, if ever, considered. In Papua New Guinea (PNG), a potential exporter of the commercial<br />
banana varieties Cavendish and Lady Finger, the endemic banana fly, Bactrocera musae<br />
(Tryon), is considered a major pest. As part of developing a PRA for PNG bananas, we studied<br />
the abundance, distribution, host use and pest status of B. musae.<br />
Methods: Distribution and abundance data for B. musae in PNG was generated through an<br />
intensive trapping program. Abundance data was correlated with rainfall, altitude and the<br />
importance of banana in the local cropping system. Host preference and larval performance of<br />
B. musae on the commercial varieties “Cavendish” and “Lady Finger” were examined in the<br />
laboratory. Field infestation of Cavendish in PNG was examined through a national fruit<br />
survey covering four agricultural production areas and bananas picked at the mature green<br />
stage of ripeness.<br />
Results: Banana fly was found to be common and widespread throughout PNG. Abundance<br />
was not significantly correlated with rainfall (R 2 = -0.02, p = 0.28) or altitude (R 2 = -0.01, p =<br />
0.35), but was weakly positively correlated with banana’s importance in the local cropping<br />
system (when presented as a percentage of local crop grown) (R 2 = 0.08, p = 0.02). Despite the<br />
fly’s high field abundance, Cavendish bananas picked from unmanaged plots around PNG<br />
were essentially unaffected by banana fly. From 111 samples, constituting 381kg of fruit and<br />
2,700 fingers, only 11 fingers were infested (0.41% infestation). Eighty-seven percent of flies<br />
reared (n = 245) came from only three fingers (0.1% of sample). Laboratory studies reflected<br />
field data, particularly for adult oviposition trials: while there was 20% adult emergence from<br />
Cavendish and Lady Finger fruit inoculated with B. musae eggs, very few flies laid eggs into<br />
fruit of either variety.<br />
Conclusions: Under laboratory conditions, the commercial banana varieties Cavendish and<br />
Lady Finger are infrequent hosts of B. musae. Cavendish is also a rare field host and we<br />
suspect the same is true for Lady Finger. In PNG, banana fly infests over-ripe bananas, as well<br />
as some sweet native varieties (authors’ unpublished data), but not the main commercial<br />
varieties when picked at the mature green stage. This project reinforces the importance of<br />
detailed host use studies for developing PRAs, rather than relying on generic literature records.<br />
Keywords: Pest risk analysis; agricultural production, abundance and distribution, host use.<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
INSPECTION OF ORGANIC PRODUCTS BY DETECTOR DOGS.<br />
Merayo, Eduardo*; Rosillo, Mario & Gobet, Luis.<br />
Fundación Barrera Zoofitosanitaria Patagonica FunBaPa- Programa Incan Trehua. Email:<br />
emerayo@funbapa.org.ar<br />
Background: The Incan Trehua is a Program developed by FunBaPa, Iscamen and Senasa. This<br />
program is aimed at training dogs for the detection of animal and vegetable derived products.<br />
These banned substances should not enter to the protect product areas due to sanitary reasons.<br />
The Incan Trehua program means “protector dogs” in Mapuche language and it`s pioneering in<br />
Latin America. The aim of this program is to enhance the perfomance of the Quarantine<br />
Patagonian System. By use of the olfactory sense to detect loads with restrictic admissions<br />
products. Incan Trehua improves the effectivenness in the sanitary control stations and making<br />
the inspectors work easier.<br />
Methods: Beagle and Labrador retriever are the chosen dog breeds for this project. The training<br />
period last a mínimum of six weeks, during that it stimulates the Memory of Olfactory Fields<br />
of the dog. This is archieved trough sterilized lints impregnated in contact with differents fruits<br />
and vegetables products and associated to many acts that the dog does, what its called<br />
“Associative Kits”<br />
Results & Conclusions: The Incan Trehua program extends its services throughout the most<br />
importans land points of control, spread over the provinces of Buenos Aires, Neuquen, La<br />
Pampa, Rio Negro and Mendoza, the Neuquen International Airport and the Viedma National<br />
Airport, protecting the entry of banned fruit, vegetables and animal origin products to<br />
Patagonia. Throughout the six years the program has, we achieved and statistic of, in a hundred<br />
vehicles controled by dogs, they detect banned organic products in ten of them. This takes over<br />
the statistics of man detection and provide a more friendly and accepted way of search as well.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
THE MEDFLY, CERATITIS CAPITATA (WIEDEMANN), AS VECTOR OF ERWINIA<br />
AMYLOVORA, THE CAUSAL AGENT OF FIRE BLIGHT IN ROSACEOUS PLANTS.<br />
Ordax, Mónica 1 ; Piquer-Salcedo, Jaime E. 1 ; Sabater-Muñoz, Beatriz 2* ; Biosca, E G. 3 ;<br />
López, María M. 1 & Marco-Noales, Ester 1 .<br />
1 Instituto Valenciano de Investigaciones Agrarias (IVIA), Centro de Protección Vegetal y Biotecnología, Ctra. de<br />
Moncada a Náquera km. 4,5; Moncada 46113- Valencia (Spain); 2 Unidad Asociada de Entomología IVIA– CIB<br />
CSIC, Centro de Protección Vegetal y Biotecnología, IVIA. Email: sabater_bea@gva.es; 3 Dpto. Microbiología y<br />
Ecología, Universidad de Valencia, 46100, Burjassot, Valencia.<br />
Background: The bacterium Erwinia amylovora is the causal agent of fire blight in rosaceous<br />
plants, and is one of the most important pests on the EPPO A2 list, being under quarantine<br />
measures for plant movement. It has been widely accepted that fruits from fireblight affected<br />
countries present an insignificant risk of transmission in fire blight free countries, despite the<br />
recent description of the viable but non-culturable (VBNC) condition, a bacterial physiological<br />
state that is undetectable by cultural methods. Till this moment, only sap-sucking insects or<br />
pollinators had been implied in fire-blight transmission. The Mediterranean fruit fly, Ceratitis<br />
capitata (Wiedemann) (Diptera: Tephritidae), is a devastating fruit pest worldwide. Besides the<br />
direct damage caused to fruit production and export, the medfly also poses a serious indirect<br />
threat to the plant health by acting as plant pathogen vector. This work describes the E.<br />
amylovora transmission ability of medfly and its implication on fruit trade.<br />
Methods: Five days old individuals of IVIA2002 strain were used to determine: i) its<br />
transmission ability and ii) the survival of E. amylovora. For the transmission ability, medflies<br />
were allowed to feed on contaminated mature apples for 48h and then other healthy apples or<br />
pear sprouts were challenged to detect transmission during 5-7days. For E. amylovora survival<br />
assays, medflies were put in contact with contaminated mature apples for 48h, and then tested<br />
for E. amylovora presence at 7, 14, 21 and 28 days after removing apples. The detection of the<br />
plant pathogen was determined in bacterial CCT medium and by PCR, both from plant material<br />
and medflies.<br />
Results: The medfly was able to transmit E. amylovora to fruit peel or pulp after 5 days<br />
exposition at 10 6 -10 3 cfu, respectively. The medfly induced fireblight symptoms in pear sprouts<br />
after 5-7 days of transmission. We were able to isolate culturable E. amylovora cells from<br />
medfly after 7 or 14 days from exposition (10 3 -10 6 cfu/medfly). However, with longer<br />
bacterium-medfly contact periods, the bacterium adopted the VBNC state. Medfly extracts (21<br />
or 28 days old) containing these VBNC cells, induced fireblight symptoms in fruits and pear<br />
sprouts and tested positive by PCR..<br />
Conclusions: This is the first time that the medfly is assessed as a vector of a plant pathogen<br />
such as E. amylovora. In this host the bacterium maintains its pathogenic potential during<br />
almost one month, being transmitted in high numbers to healthy plant material.<br />
Keywords: Medfly, Erwinia amylovora, fire blight, transmission, survival, plant pathogen vector.<br />
211
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GEOGRAPHICAL DISTRIBUTION OF ANASTREPHA GRANDIS (MACQUART) IN THE<br />
STATE OF SÃO PAULO, BRAZIL.<br />
de Souza-Filho, Miguel F.* 1 ; Raga, Adalton 1 ; Montes, Sônia M. N. M. 2 ; Azevedo-Filho,<br />
Joaquim A. de 3 ; Garcia, Maria J. De M. 4 & Almeida, Aparecida M. de 4 .<br />
1 Instituto Biológico, Caixa Postal 70, CEP 13012-970, Campinas, São Paulo (Brazil). Email:<br />
miguelf@biologico.sp.gov.br. 2 APTA Regional - Pólo Alta Sorocabana, Presidente Prudente, São Paulo, Brazil.<br />
3 APTA Regional - Pólo Leste Paulista, Monte Alegre do Sul, São Paulo, Brazil. 4 APTA Regional - Pólo Centro<br />
Oeste, Bauru, São Paulo, Brazil.<br />
Background: Anastrepha grandis is considered a fruit fly species of quarantine importance and<br />
it has been registered in the south, southeast and central of Brazil. In the State of São Paulo is<br />
concentrated the highest volume of cucurbitaceae trade for Brazilian market. The objective of<br />
the present study was to investigate the geographical distribution of A. grandis in the State of<br />
São Paulo.<br />
Methods: The occurrence of A. grandis was based on the literature, such as theses, articles and<br />
reports from the congress. Additional informations were based on field surveys conducted from<br />
2009 to 2010. In the both cases, we considered collections of A. grandis from fruit hosts and<br />
monitoring studies in many crops.<br />
Results: A. grandis is reported in 22 municipalities of northeast, central, southwest and east of<br />
the State of São Paulo. Eleven municipalities are located in the east region.<br />
Conclusions: No specimen of A. grandis was collected from northwest and extreme west<br />
regions of the State of São Paulo, which it was included the squash production areas for<br />
exportation.<br />
Keywords: South American Cucurbit Fruit Fly, occurrence, localities, quarantine pest, Cucurbitaceae
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
THE THREAT OF BACTROCERA INVADENS DREW TSURUTA & WHITE, TO SOUTH<br />
AFRICA: SURVEILLANCE, CONTINGENCY AND TRADE RESPONSE.<br />
Venter, Jan Hendrik* 1 ; Manrakhan, Aruna 2 ; Matebe, Yolanda 1 ; Brown, Leslie 3 &<br />
Stones, Wilna 4<br />
1 Department of Agriculture Forestery and Fisheries, (DAFF), Private Bag X14, Pretoria, South Africa, 0031,<br />
janhendrikv@daff.gov.za; 2 Citrus Research International (CRI), Nelspruit, South Africa; 3 DFPT Research,<br />
Stellenbosch, South Africa; 4 Subtropical Fruit Growers Association, Tzaneen, South Africa.<br />
Background: Exotic fruit flies remain a major concern to South African fruit producers aiming<br />
to reach the international market. Not only do exotic flies pose a risk to fruit production, but<br />
there is a real threat that export markets may be lost or at least that exports could be<br />
temporarily suspended should new species be introduced to the country. Bactrocera invadens,<br />
an invasive fruit fly of Asian origin, was first detected in Africa in 2003 in Kenya. Thereafter,<br />
the fly was reported in several other African countries including those in southern Africa such<br />
as Zambia, Mozambique and Namibia. B. invadens males are strongly attracted to methyl<br />
eugenol which can be used for monitoring of this fly. Since 2006, South Africa has been<br />
conducting a fruit fly surveillance programme through collaboration between the National<br />
Plant Protection Organisation (NPPO), fruit industries and research organizations for early<br />
detection of exotic fruit flies including B. invadens. Contingency plans specific to B. invadens<br />
have also been put in place in South Africa and these include the setting up of a national<br />
steering committee on B. invadens and the drafting of an action plan specific to the Invader fly<br />
where response actions for initial incursions and longer term management strategies are<br />
described. Import conditions for fresh fruit have been revised, which has had an impact on<br />
trade of certain host commodities from countries where B. invadens is present.<br />
Methods: The surveillance network for detection of B. invadens currently consists of 616<br />
methyl eugenol baited traps and 136 BioLure 3 component baited traps. Traps are deployed in<br />
major towns, ports, airports, high risk border areas as well as in production areas. Traps are<br />
serviced at least once a month. The trapping information from the NPPO and fruit industries is<br />
then captured in a centralized database.<br />
Results & conclusions: To date, South Africa remains free from B. invadens. Should there be<br />
an incursion of B. invadens in South Africa, a stockpile of emergency materials for use<br />
according to the relevant action plan, will be available immediately for eradicating this pest.<br />
Keywords: Invader fruit fly, detection, action plan, trade implications<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PATAGONIAN QUARANTINE SYSTEM.<br />
Villalba, Daniel Nestor*; Justo, Adrian D. & Merayo, Eduardo M.<br />
Patagonian zoophytosanitary barrier foundation. Email: dvillalba@funbapa.org.ar<br />
Background: The Patagonian Protected Region, which is the southernmost area in South<br />
America, covers an area of 835.000 km2. Its natural boundaries are the Andean Range to the<br />
west, the Atlantic Ocean to the east and the Colorado and Barrancas Rivers, along which the<br />
terrestrial control posts in every way-in to the protected area are settled, to the north. The<br />
Patagonian Quarantine System is conducted and administrated by the Patagonian<br />
Zoophytosanitary Barrier Foundation (FUNBAPA), a non governmental organization where<br />
institutions coming from both the public and private sectors, like SENASA, regional provinces<br />
administrations, and growers and breeders associations, take part. It is a joint organizational<br />
model of public and private sectors, non-profit and with an active participation of its<br />
components. The aim of the Patagonian Quarantine System is the supervision of the way-in<br />
and way-out of plant or animal originated products by the enforcement of federal sanitary<br />
regulations in order to avoid the introduction of pests and/or diseases that could affect the local<br />
production. Legislation applied through the Patagonian Quarantine System is federal and is<br />
originated both in the Ministry of agriculture, livestock and fisheries of the nation (MAGyP)<br />
and the Agroalimentary Quality and Health National Service (SENASA).-<br />
Methods: The Patagonian Quarantine System involves 34 terrestrial inspection posts 13<br />
airports, 1 railroad control post and all the seaports of the Patagonian coast. Terrestrial posts<br />
work all the year round, 24 hours a day, and in each post the number of inspectors is directly<br />
related to the number of vehicles that pass along. More than three hundred people among<br />
professionals, technicians and inspectors are involved, being trained and evaluated on an<br />
ongoing basis. Supervision and quality control is carried out by the own managers staff and the<br />
audits are carried out by SENASA. Inspection work is complemented by the FUNBAPA Incan<br />
Trehua Program which uses dogs specially trained for the detection of organic products.<br />
Besides, special equipment for the same purpose is used at airports so as to improve and<br />
facilitate inspection tasks. The financing of the Patagonian Quarantine System is given by the<br />
funds originating from the tariff control, inspection and disinsectation is levied in terrestrial<br />
barrier checkpoints, contributions from the Government and the provinces.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
POTENTIAL DISTRIBUTION OF CERATITIS ANONAE IN CHINA BASED ON<br />
PREDICTION OF ECOLOGICAL NICHE MODELS.<br />
Zhang, Run-Jie* & Li, Bai-Ni<br />
State Key Laboratory for Biocontrol & Institute of Entomology, Sun Yat-Sen University, Guangzhou 510275,<br />
P.R. China. E-mail: lsszrj@mail.sysu.edu.cn.<br />
Backgrounds: The Ceratitis anonae (Graham) is native to the Afrotropical region. It is a<br />
polyphagous species with wide host ranges including almond, annonaceous, avocado, cherry,<br />
coffee, guava, mango, strawberry, and so on. This insect pest is an important quarantine<br />
invasive pest for China and its larva are continually intercepted from fruits carried by incoming<br />
passengers at Guangdong ports. The potential geographic distribution of this species in China<br />
is of considerable concern in terms of biosecurity.<br />
Methods: In this study, three ecological modeling methods (ENFA: ecological niche factor<br />
analysis, Mahalanobis Typicality, and Maxent: maximum entropy niche-based modeling) were<br />
used to predict its potential distribution. Native range of C. anonae was derived from the Royal<br />
Museum for Central Africa and the Global Biodiversity Information Facility. Records of<br />
occurrence were obtained from the literature and electronic resources. In total, 144 unique<br />
records were obtained for C. anonae.<br />
Results: The results show that Maxent has the best prediction performance, followed by<br />
Mahalanobis Typicality, and ENFA. The difference of prediction capabilities between Maxent<br />
and Mahalanobis Typicality is not significant. Based on prediction outcome of Maxent,<br />
suitable areas of C. anonae in China are limited in Guangxi, Guangdong, Hainan, and a few<br />
areas of Yunnan. The probability of C. anonae permanently establishing in southern China<br />
exist indeed but small. Jackknife analysis indicates that six variables have notable influences<br />
on the distribution pattern of C. anonae, i.e. ground frost frequency, annual mean precipitation,<br />
precipitation of October, precipitation of April, annual minimum temperature, and vapor<br />
pressure. More researches on the influence of environmental variables on survival of C. anonae<br />
are needed to determine its suitable distribution area.<br />
Conclusions: The probability of C. anonae permanently establishing in southern China exist<br />
indeed but small. Six environmental variables have notable influences on the distribution<br />
prediction of C. anonae, and further researches about their influence on survival of C. anonae<br />
are needed.<br />
Keywords: Ceratitis anonae; ENFA; Mahalanobis Typicality; Maxent; potential distribution prediction<br />
215
Session 5<br />
SIT Principles & Application
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
OLIVE FLY: FROM SMALL SCALE PRODUCTION TO MASS-REARING.<br />
Ahmad, Sohel*, Wornoayporn, Viwat, Haq, Ihsan ul, Cáceres, Carlos & Jessup, Andrew<br />
J.<br />
Insect Pest Control Laboratory, Joint FAO/IAEA Agriculture and Biotechnology Laboratories. A-2444,<br />
Seibersdorf, Austria. Email: S.Ahmad@iaea.org<br />
Background: The olive fly, Bactrocera oleae (Diptera: Tephritidae), is considered an ideal<br />
candidate for control by the sterile insect technique (SIT) as part of an integrated pest<br />
management approach because it attacks only olives and does not disperse naturally great<br />
distances unlike other Bactrocera spp. One major constraint in the development of a successful<br />
and cost-effective SIT programme for olive fly is the large scale production of high quality<br />
mass-reared flies. The aim of this work was to develop cost-effective methods for massrearing.<br />
Methods: The following three basic parameters in mass-rearing were examined: Cage density<br />
of adult flies, size and design of the adult oviposition cage, and egg collection methods.<br />
Results: The results showed that an adult fly density of 4.1 cm 2 (internal surface area of cage)<br />
per fly in a medium-sized cage (0.036m 3 ) produced up to 11.5 eggs/female/day over the life of<br />
flies. Further research on other cage sizes ranging from 0.015 m 3 to 0.4 m 3 resulted in egg<br />
volumes from only 2.8 to 6.8 eggs/female/day. Our newly developed method of egg collection<br />
using a flat egging panel proved cost-effective, more efficient, and enabled us to elevate the<br />
colony size to a level that mass-production can be started.<br />
Conclusions: As a consequence of this work, we are now rearing the flies in large cages (0.4<br />
m 3 ) formerly used for mass rearing of Mediterranean fruit fly and we are now optimizing the<br />
density of olive fly in cages that are more acceptable for cost-effective mass rearing of this<br />
insect.<br />
Keywords: Olive fly, fly density, cage design, egg production<br />
219
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
STERILE INSECT TECHNIQUE FOR CONTROLLING CUCURBITS FRUIT FLY, DACUS<br />
CILIATUS (LOEW) (DIPTERA:TEPHRITIDAE).<br />
Al-Taweel, Ayad A. 1* ; Al-Shammary, A.J. 1 & Ahmed, R.F. 2<br />
1 IPC Center, Ministry of Science & Technology, Baghdad, Iraq. 2 College of Agriculture, Baghdad University,<br />
Baghdad, Iraq.<br />
Background: The cucurbit fruit fly, Dacus ciliatus (Loew) is one of the most important fruit<br />
flies on cucurbits in Iraq. The Iraqi scientist thinking to apply sterile insect technique to control<br />
this pest instead of insecticide for environment concern, after they carried out several studies<br />
regarding its biology, ecology and behavior.<br />
Methods: Different doses of irradiation were tested on 4-5 days old pupa. Egg laid was<br />
measured for each dose, using also irradiated females as control.<br />
Results: The results of this study revealed that the dose of 0.75 kGy caused complete sterility to<br />
the females and the dose of 0.90 kGy caused complete sterility to the males when they<br />
irradiated as 4-5 days old pupa. The oviposition was recorded in the mating between<br />
unirradiated males x irradiated females (1); irradiated males x unirradiated females (2) and<br />
irradiated males x irradiated females (3) for the doses 0.15-60 kGy, while for the doses 0.75 &<br />
0.90 kGy no eggs laid were observed for the mating 1 & 3 only. Furthermore, the percentage of<br />
eggs hatch was zero for doses 0.75 & 0.90 kGy for the mating 1 & 3, while it was zero for<br />
mating 2 for the dose 0.90 kGy only.<br />
Conclusions: The results of this study also showed that doses of gamma rays have no effect on<br />
pupation percentage of the larvae produced from hatched eggs of irradiated males & females as<br />
pupa of five days with all doses & mated to opposite unirradiated sex or mated together.<br />
Finally the results of this study also showed that all doses have no effect on adult emergence<br />
percentage & sex ratio.<br />
Keywords: Dacus ciliatus , Gamma Rays, Fecundity, Cucurbits
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
IMPROVEMENT IN PRODUCTION AND QUALITY PARAMETERS AT NEW SIT AFRICA<br />
MEDFLY, CERATITIS CAPITATA (WIEDEMANN), REARING FACILITY.<br />
Arnolds, Luciano 1 ; Vermeulen, Nathan* 1 ; Johnson, Jerome 2 & Barnes, Brian 2<br />
1 SIT Africa (Pty) Ltd, Stellenbosch, South Africa. Email: luciano@sitafrica.co.za; 2 Technical Consultant for SIT<br />
Africa (Pty) Ltd, Stellenbosch, South Africa.<br />
Background: The South African Medfly SIT programme was initiated in 1999 when mass<br />
rearing of the tsl strain Vienna 7 was started in a small, renovated building in Stellenbosch. A<br />
number of strain changes have since taken place, and Vienna 8 has been used since 2003. In<br />
1999 the maximum production capacity was approximately 8 million sterile males per week.<br />
Production and quality was adversely affected by factors including equipment breakdown,<br />
inadequate environmental insulation, lack of space, and micro-organism growth in breeding<br />
rooms, all of which was compromised by a shortage of funding. The building was old and the<br />
general infrastructure was increasingly becoming degraded. In 2001 a new Chromodec®<br />
insulated-panel building was erected to house the adults and a quality control laboratory.<br />
Although this led to more stable production and quality, larval rearing and pupal handling in<br />
the old building, and ultimately sufficient production of high quality sterile males, remained at<br />
risk from the failing infrastructure. Roll-out of the SIT programme to new areas required an<br />
increase in production, which put further pressure on the facility. A new building to house the<br />
immature stage rearing and handling was essential.<br />
Development: In 2010, following a considerable injection of funding from government, a 480<br />
m 2 Chromodec® insulated-panel extension was added to the adult room for immature stage<br />
rearing. The capacity of this new facility is estimated to be at least 40 million sterile males per<br />
week, which will allow a far greater fruit production area to be included under the SIT blanket.<br />
Results: In the 6 months since the new building was commissioned, production has<br />
significantly stabilised, and key production and quality parameters have all increased, in some<br />
cases substantially, in comparison with the previous 12 month-period in the old building.<br />
Overall average increases in the filter, amp 1, amp 2 and release streams (in percentage points)<br />
were as follows: egg production, 40.9; efficacy, 25.7; egg hatch (48 h), 4.0; flight ability<br />
(colony females), 3.6; flight ability (colony males), 3.9. Pupal weight in the release stream<br />
increased by 2.5 percentage points, and flight ability of sterile males by 2.5. The long-term<br />
trends in all parameters from the new building show a narrower band of fluctuation than in the<br />
old building, and all daily and weekly figures are mostly on or above target. Currently, 11<br />
million sterile males are released weekly in three separate production areas, protecting a total<br />
area of nearly 24,000 ha of fruit.<br />
Conclusions: The above improvements in production and quality augur well for the planned<br />
expansion of Medfly SIT in the Western Cape and even further afield in the near future, and for<br />
the Medfly SIT programme in South Africa in general.<br />
Keywords: Medfly, mass-rearing, tsl, improved quality, SIT<br />
221
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SUITABLE LARVAL DIETS FOR ANASTREPHA FRATERCULUS (DIPTERA:<br />
TEPHRITIDAE).<br />
Braga Sobrinho, Raimundo 1* ; Guimarães, J.A 2 ; Araujo, Elton L. 3 & 4 Moreira, M.A.B 4<br />
1 Embrapa Agroindústria Tropical – Rua Dra. Sara Mesquita, 2270 - 60511-110 – Fortaleza Ceará Brasil.<br />
braga@cnpat.embrapa.br; 2 Embrapa Hortaliças – Parque Estação Biológica – PqEB – 70770-901 – Brasília DF,<br />
Brasil. 3 Universidade Federal Rural do Semiárido, BR 110, Km 47- 59625-900 – Mossoró – RN, Brazil;<br />
4 Embrapa Tabuleiros Costeiros – Av. Beira Mar, 3250. 49025-040 – Aracaju SE<br />
Background: The genus Anastrepha (Diptera:Tephritidae) is the largest and most economically<br />
important genus of true fruit flies in the American tropics and subtropics. It is indigenous to the<br />
Americas, presently with no distribution outside the Western Hemisphere. Among other<br />
species, the South American fruit fly, Anastrepha fraterculus (Wiedemann, 1830) is highly<br />
economic important in South America, especially in Brazil. Attempts to improve diet quality<br />
for mass rearing A. fraterculus have been ongoing for more than twenty years. The diets<br />
currently used throughout the world were based on combined ingredients taken in account their<br />
availability, physical properties and economics with little emphasis on nutrient value. A mass<br />
rearing system and Sterile Male Technique (SIT) for A. fraterculus are underway and will<br />
remain a challenge. A technique to mass rear A. fraterculus is the start point for the<br />
development of SIT. The objective of the present work was to identify appropriate ingredients<br />
for suitable larval and adult diets to be used as a platform to develop a protocol for mass<br />
rearing the South American fruit fly.<br />
Methods: A screening of eleven different larval diets (four replications) was performed in order<br />
to find suitable and economic diets for further comparison tests. The efficacy of the diets was<br />
determined by statistical analysis including egg hatching, pupal recovery, pupae weigh, adult<br />
emergence and adult flying ability. The general procedure for quality control tests were based<br />
on protocols specified in the International Fruit Fly Control Manual (FAO/IAEA/USDA,<br />
1998).<br />
Results: The final goal of a fruit fly mass rearing production system is a consistent result of<br />
healthy and competitive adults. This success is very dependable of high control quality of all<br />
laboratory procedures and specially a suitable and economic diet for larvae. From the results of<br />
these larval diets experiments, there is clear evidence that the source of protein is determinant<br />
for getting healthy and competitive adults. Showed by fly ability test. From six diets tested,<br />
three of them with 9.0, 10.0 and 13.0% of protein and 20.0% of cane bagasse as the bulk agent<br />
pesented the best results for egg hatching, pupal recovery, pupal weight, adult emergence and<br />
fly abilitiy when is compared with the other diets with less protein content. Since there were no<br />
statistical differences among these thee diets it is clear that is more economically feasible to<br />
recommend the diet with 9% of protein.<br />
Conclusions: It is clear that protein in diet larvae improves larval development, pupal recovery<br />
and adult fly ability. Larval diets with 9.0% of protein show to be adequate for a mass rearing<br />
program with focus on SIT. The bulk agent sugar cane bagasse improves larval feeding by<br />
facilitating the larvae movement though out the diet mass.<br />
Keywords: South American fruit fly, SIT, pupae, mating, fly ability
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
INFLUENCE OF PRERELEASE DIET TO BACTROCERA DORSALIS (HENDEL) AND<br />
BACTROCERA CORRECTA (BEZZI) STERILE MALES ON MATING PAIRS.<br />
Chinvinijkul, Suksom*; Pinkaew, Supaap & Orankanok, Watchreeporn<br />
Department of Agricultural Extension, 2143/1 Paholyothin Road, Chatuchak, Bangkok 10900 Thailand. Email:<br />
chinvinijkuls@yahoo.com.au.<br />
Background: Released sterile males fed only water-agar are currently applied in fruit fly<br />
control project area in Thailand. In general, effectiveness of protein was shown to the sexual<br />
maturity in fruit fly males for the mating capability. We reported here on influence of<br />
prerelease diet of sterile males on mating pairs which indicate their successful of mating<br />
performance.<br />
Methods: Sexing separation of the 53th generations of Bactrocera dorsalis (Hendel) and the<br />
55th generations of Bactrocera correcta (Bezzi) sterile mass reared males were conducted<br />
within 24 hours of eclosion. Five groups of neonate fruit flies were fed with water-agar only,<br />
sugar (S), sugar and protein hydrolysate ratio 3:1 w/w (SP = 3:1), sugar and protein<br />
hydrolysate ratio 2:1 w/w (SP = 2:1) and protein hydrosate (P) only for two days. Then each<br />
group were given a standard adult diet SP (3:1) and water throughout the experiment. During<br />
15 testing days of B. dorsalis, each 2+n day-old (n=1-15) of 50 sterile males fed with<br />
difference type of diet were mated with 50 virgin mass reared females, 12 day-old. With the<br />
same procedure during 18 testing days of B. correcta, each 2+n day-old (n=1-18) of 50 sterile<br />
males fed with difference type of diet were mated with 50 virgin mass reared females, 17 dayold.<br />
Results: From the 7th-15th testing days after prerelease diet feeding, young sterile B. dorsalis<br />
and B. correcta fed with S, SP = 3:1, SP = 2:1 and protein hydrolysate showed higher different<br />
mating percentage than those fed only water-agar significantly. Total testing period (15 days<br />
for B. dorsalis and 18 days for B. correcta) percentage of mating pairs of sterile male fed diet<br />
formula were highly significant difference than other fed with water-agar only. Young sterile<br />
male of B. dorsalis fed protein hydrolysate showed the highest percentage of mating pairs but<br />
not significantly difference to those fed S, SP = 3:1; and SP = 2:1. Bactrocera correcta fed<br />
with SP=3:1 achieved the highest percentage of mating pairs and significantly higher than<br />
sterile males fed with S and P only but not significantly difference to sterile males fed SP =<br />
2:1.<br />
Conclusions: Prerelease diet, sugar only, combination of sugar and protein hydrolysate 3:1, 2:1<br />
and portein hydrolysate only showed non significantly difference of mating pairs for sterile B.<br />
dorsalis males but B. correcta males fed with SP=3:1 and SP=2:1 showed mating percentage<br />
significantly superior than S and P feeding. Sterile males of B. dorsalis and B. correcta fed<br />
only water-agar as prerelease diet obtained very low mating success.<br />
Keywords: Bactrocera dorsalis, Bactrocera correcta, young sterile, prerelease diet, mating pairs<br />
223
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
STARTER DIET AS INITIATING FEED FOR THE ANASTREPHA LUDENS LARVAE IN<br />
MASS REARING CONDITIONS IN MOSCAFRUT FACILITY.<br />
Domínguez, Julio; Hernández, M. Refugio; Solís E., Eduardo A. * ; Rovelo M., Rosa E. &<br />
Báez M., Raúl<br />
Planta Moscafrut, Camino a los Cacahoatales S/N. Metapa de Domínguez, Chiapas, México. C.P. 30860.<br />
Background: The nutritional conditions in insects have a significant role in the ability of males<br />
to attract females and result in mating. The relation between nutrition and male reproductive<br />
behavior is shown by production of signs for courtship and males may use nutrients to<br />
synthesize materials for production of pheromones or substances transferred during copulation<br />
such as sperm (Braga, R. 2006). There is a general agreement that protein-fed males are more<br />
likely to mate. Vienna-7 sterile Med fly males mate much more frequently in cages with<br />
protein than did the regular sugar fed colony. On the other hand wild males that emerged in the<br />
laboratory and were tested for mate in a field cage also succeeded significantly better than<br />
sugar fed males (Braga, R. 2006). For Moscafrut Facility is important to maintain the<br />
characteristics of the flies and improve them at the level with the wild ones, so trying the gelid<br />
food as starter diet for the mass rearing to rise Anastrepha ludens and A. obliqua is expected<br />
that it will give the necessary nutriments which the larvae requires, transferring to the final diet<br />
(the traditional diet in less proportion) to finish its maturity process; giving the desire mass<br />
rearing and quality parameters and in the other hand the high costs of the elaborating diet will<br />
be reduced (there is a high consume of imported ingredients for the traditional diet).<br />
Method: After seeding the egg in the jelly diet (Nutryfly®) the new larvae was transferred to<br />
the normal diet to finish its development, then it was separated from the diet, continuing with<br />
its recovery as pupae and adult. So the parameters that will be tested are: yield egg-larvae,<br />
number of larvae per tray and larvae weight. The values of established parameters are: Yield<br />
egg-larvae (Y.E-L): 75%; weight larvae: 26 mg; number larvae/tray: 20,900.<br />
Results: From the three egg densities (1.2 ml, 1.4 ml and 1.6 ml) the results were: 1.2 ml:<br />
73.36% Y.E-L, 25.78 mg and 19,273 larvae/tray; 1.4 ml: 66.88 Y.E-L, 25.14 mg and 20,504<br />
larvae/tray; 1.6 ml: 58.86% T.E-L, 24.39 mg and 20,578 larvae/tray. With the data obtained<br />
from the 1.2 ml was selected for a second phase to improve it, this because two of the three<br />
parameters were close to get, so in the second part the test was divided, a seeding was done in<br />
the morning (9:00 am) and the other in the afternoon (6:00 pm) considering the egg hatching;<br />
the results were: 1.2 ml in the morning: 69.47% T.E-L, 28.37 mg and 18,211 larvae/tray; 1.2<br />
ml in the afternoon: 75.59% T.E-L, 27.80 mg and 19,958 larvae/tray.<br />
Conclusions: according the results using a low density (1.2ml) it is possible to obtain the<br />
production and the quality parameters (even the yield is high and the longevity per cent); if the<br />
temperature for the egg hatching (increasing it) can be regulated is possible to improve the<br />
results in the starter feed; so it is useful to try 1.2 ml for the further tests between the gelid food<br />
as starting feed with the traditional feed and compare the results in a mass rearing system in A.<br />
ludens, also for A. obliqua. Finally other benefits come from this kind of feeding, like the<br />
residual reduction of the diet, the minimized import ingredients consumption, the space and the<br />
storage are optimized in the areas.<br />
Keywords: starter diet, yield egg-larvae, larvae weight and larvae per tray
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EFFECT OF ANOXIA ON MASS REARED QUEENSLAND FRUIT FLY BACTROCERA<br />
TRYONI (FROGGATT) (DIPTERA: TEPHRITIDAE).<br />
Dominiak, Bernard C. 1 , Sundaralingam, S. 2 , Jiang, L. 2, , & Nicol. H. I. 3<br />
1 NSW Department of Industry and Investment, Locked Bag 21, Orange, New South Wales, 2800, Australia.<br />
Email: bernie.dominiak@industry.nsw.gov.au; and The Department of Brain, Behaviour & Evolution, Macquarie<br />
University, NSW 2109, Australia; 2 Elizabeth Macarthur Agricultural Institute, NSW Department of Industry and<br />
Investment, PMB 8, Camden, New South Wales, 2570, Australia; 3 Nicol Consulting, 95 Ophir Road, Orange,<br />
New South Wales, 2800, Australia<br />
Background: Area wide treatments using the Sterile Insect Technique (SIT) rely on irradiated<br />
or sterile fruit flies to produce no viable offspring in wild fly populations. Irradiation of<br />
oxygenated tissues is known to cause adverse effects on fruit fly and need to be minimised. A<br />
simplest method to decrease the level of oxygen in pupae is to seal pupae in plastic bags and<br />
allow the oxygen levels to decline to the low levels. However some species do not survive<br />
well in depleted oxygen for long periods. We report here on tests conducted on Bactrocera<br />
tryoni (Froggatt) Qfly to evaluate the adverse effects of different periods of anoxia.<br />
Methods: Four experiments were conducted using irradiated and non irradiated Qfly pupae at<br />
17°C, 21°C and 25°C. Anoxia for 56, 96, 168 and 192 hours were tested. At the end of each<br />
period of anoxia, the bags were opened and eclosion allowed to occur. After full eclosion, the<br />
percentage successful eclosion was assessed and analysed statistically.<br />
Results: In the first experiment using unirradiated pupae for up to 56 hours of anoxia, the<br />
highest average eclosion of 89.5% occurred at 21°C. In the second experiment using<br />
unirradiated pupae at up to 96 hours of anoxia, there was a decline from 89.4% eclosion at 0<br />
hours to 78.1% at 96 hours with again 21°C resulting in higher eclosion. There were small<br />
changes in the percentage of partially eclosed adults but larger increases in uneclosed pupae. In<br />
the third experiment, irradiated and unirradiated pupae were compared up to 192 hours of<br />
anoxia at only 17°C. The average eclosion declined by 14% to about 70%. Unirradiated pupae<br />
had 8% higher average eclosion compared with irradiated pupae. The fourth experiment used<br />
only irradiated pupae at 17°C. Full eclosion at 0 hours was 78.0% and declined to 64.6% at 192<br />
hours.<br />
Conclusions: Optimum eclosion appears to occur at 21°C. Ecolsion declined at approximately<br />
0.1% eclosion per hour of anoxia during the first 96 hours. There was a 12-14% decline in<br />
eclosion after 192 hours of anoxia. These results are at variance with many other species. Qfly<br />
are tolerant of long periods of anoxia and current practices will not disadvantage eclosion rates<br />
as long as temperatures are kept at optimal.<br />
225
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
CAN STERILE SPERM PRESENCE AND FRUIT DAMAGE BE USED AS INDICATORS OF<br />
EFFICACY OF MEDFLY SIT PROGRAMS?<br />
Juan-Blasco, María 1* ; Sabater-Muñoz, Beatriz 1 ; Martínez-Utrillas, Miguel Ángel 2 ;<br />
Martínez, Marta 2 ; Plá, Ignacio 2 ; Argilés, Rafael 2 ; Castañera, Pedro 1 ; Jacas, Josep A. 3 &<br />
Urbaneja, Alberto 1 .<br />
1 Unidad Asociada de Entomología UJI-IVIA-CIB CSIC. Centro de Protección Vegetal y Biotecnología; Instituto<br />
Valenciano de Investigaciones Agrarias, IVIA; Ctra. de Moncada a Náquera km 4.5; E-46113-Moncada (Spain).<br />
Email: juan_marbla@gva.es. 2 Departamento de Plagas. TRAGSA. Release Unit. Ctra. de Moncada a Náquera km<br />
4.5; E-46113-Moncada (Spain). 3 Unidad Asociada de Entomología UJI-IVIA-CIB CSIC. Departament de<br />
Ciències Agràries i del Medi Natural; Universitat Jaume I, UJI; Campus del Riu Sec; E-12071-Castelló de la<br />
Plana (Spain).<br />
Background: A Sterile Insect Technique (SIT) programme against the Mediterranean fruit fly,<br />
Ceratitis capitata, has the goal of decreasing the target population by area-wide release of large<br />
numbers of sterile males. These sterile males will mate with wild females, which will then<br />
produce unviable eggs and consequently a decrease of the wild population. Success of a SIT<br />
programme requires a continued evaluation of its efficacy which is currently evaluated by<br />
monitoring of both wild and sterile males by means of pheromone baited traps. In this work,<br />
the efficacy in suppression of 5 medfly male ratios (wild:sterile) were evaluated in a citrus field<br />
experiment. For this aim, the correlation between sterile males mate achievement, measured by<br />
molecular sperm identification in trapped wild females, and the production of viable progeny<br />
measured as pupae emerged from sentinel fruits, was obtained.<br />
Methods: The experiment was conducted in 17-y-old single mesh-caged clementine trees. In<br />
each cage (3 x 3 x 3 m) 50 wild males and 50 wild females with the corresponding sterile<br />
males (ratios of 1:0; 1:1; 1:5; 1:10 and 1:20 wild: sterile) were released. Ten susceptible fruits<br />
were hold on each tree to count after exposition the number of emerging medfly pupae per<br />
fruit. The presence of Vienna-8 sterile male sperm in the spermathecae of collected female was<br />
detected by specific PCR markers.<br />
Results: The number of medfly pupae emerged per fruit was significantly different between<br />
treatments. Treatments of 1:5 and higher ratios showed a number of pupae emerged<br />
significantly lower than treatments 1:1 and 1:0 where no sterile males were released. The<br />
percentage of population reduction was around 80% for the 1:5; 1:10 and 1:20 (wild: sterile)<br />
treatments when compared to trees with no sterile males releases. Accordingly, percentages<br />
females with presence of sterile sperm were detected in those treatments where sterile males<br />
were released. Among ratios tested, number of pupae emerged per fruit was related to<br />
percentage of sterile sperm detection.<br />
Conclusions: The 1:5, 1:10 and 1:20 (wild: sterile) C. capitata males ratios effectively reduced<br />
medfly populations and this was reflected in a reduction in fruit damage. The results of the<br />
experience showed a relation between fruit damage and sterile sperm molecular identification.<br />
Implications of these results for a SIT programme are discussed.<br />
Keywords: Ceratitis capitata, sterile insect technique, (wild: sterile) male ratio, medfly infestation, mating<br />
success, PCR sperm detection.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
ADVANCES IN THE MASS REARING OF ANASTREPHA SERPENTINA (WIEDEMANN)<br />
(DIPTERA: TEPHRITIDAE) IN PERU.<br />
López, Karim*; Nolazco, Norma & García, Juliana<br />
Desarrollo de Métodos (DM) de la Unidad del Centro de Producción de Moscas de la Fruta (UCPMF), del<br />
Servicio Nacional de Sanidad Agraria (SENASA), Lima-Perú. Email: klopez@senasa.gob.pe.<br />
Background: Agricultural activity in the Peru has acquired great economic importance since it<br />
generates foreign exchange through export; this activity has been limited by the presence of<br />
fruit flies of the genus Anastrepha. In the Peru, one of the three species of major economic and<br />
quarantine importance is Anastrepha serpentina which causes strong damage to fruit species<br />
such as Lucuma obovata and Sapotaceae other, causing serious economic losses in the<br />
Peruvian farmers. Therefore, in the Laboratory of Methods Development SENASA is carry<br />
research related to colonization, mass rearing, sterilization behavior and fruit fly Anastrepha<br />
complex with the objective of implementing the Sterile Insect Technique (SIT) to control this<br />
pest. Here is the advance in mass rearing A. serpentina achieved in Peru.<br />
Methods: The colony established and maintained in the laboratory was obtained from pupae<br />
recovered from infested fruits from the field (of two places: Cajamarca and Ayacucho). Having<br />
obtained the adult flies, the first problem was to obtain the eggs of A. serpentina form<br />
artificially. We tested different panels of oviposition (two colors mesh Bombay). Once<br />
approved this panel, were tested for hatching of eggs and artificial diet was developed to carry<br />
out the life cycle of flies in the laboratory.<br />
Results: It was possible to establish the environmental conditions for this species, temperatures<br />
and humidity of the environments used for each stage of A. serpentina ranged from 19-28º C<br />
and 60-85% relative humidity. From the 5th generation began to collect eggs in the oviposition<br />
panel: Bombay blue mesh recommended by other researchers and as the cages have been used,<br />
oviposition increased, reaching up to 1.5 ml collecting egg per day from 4 cages with an<br />
average of 32 000 adults in each cage, from infested fruits in the laboratory. The artificial diet<br />
tested was composed of: hydrolysate enzimatic protein, sugar, methylparaben, sodium<br />
benzoate, citric acid and “chala” (leaves and stems of corn dry and milled) as texturizing agent,<br />
we were able obtained pupae with a weight average of 15 mg and a 70% of emergency.<br />
Conclusions: It is possible the progressive adaptation of A. serpentina under laboratory<br />
conditions for subsequent mass rearing in our country. The selected artificial diet is still in<br />
testing because it requires fine-tune the quantities of inputs to use and conduct more tests with<br />
other types of inputs to achieve a greater recovery of larvae and adults.<br />
Keywords: Anastrepha serpentina, fruit fly, oviposition, Sterile Insect Technique<br />
227
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PRE-RELEASE PROTEIN DIET, ALSO A KISS OF DEATH FOR ANASTREPHA LUDENS?<br />
Meza, J. Salvador 1,2* , Tejeda, Marco T. 1 , Díaz-Fleischer, Francisco 1 & Pérez-Staples,<br />
Diana 1<br />
1 INBIOTECA, Universidad Veracruzana, Apartado Postal 250, Xalapa, Veracruz, Mexico; Email:<br />
mezajs@hotmail.com; 2 National Campaign Fruit Flies. Programa Moscafrut. Acuerdo SAGARPA-IICA, México.<br />
Central Poniente No. 14 Col Centro CP 30700 Tapachula Chiapas México.<br />
Background: Protein supplements offered during the pre-release period have been shown to<br />
improve male mating competitiveness in Ceratitis capitata and Bactrocera tryoni. However,<br />
protein during pre-release could be a “kiss of death” for flies if they are released into a stressful<br />
environment where they may starve if food is scarce. Here, we tested the sexual performance<br />
and longevity of sterile Anastrepha ludens (aka Mex fly) males and females fed sugar and<br />
protein (hydrolysed yeast) during different days during the six-day pre-release period and then<br />
starved until sexual maturity.<br />
Methods: Sterile males and females were offered protein and sugar for varying numbers of<br />
days during the pre-release period for A. ludens. Protein was offered at the beginning of<br />
emergence or before release in a total of 10 combinations. After 5 days, all flies where starved<br />
and offered water only until 10 days of age. Control flies were fed continuous protein and<br />
sugar or sugar only until 10 days of age. Daily survival was noted. The number of copulations<br />
of males and females from each treatment were recorded for flies that survived until 10 days of<br />
age.<br />
Results: Preliminary results suggest that flies fed on sugar and protein for 5 to 3 days and then<br />
offered water only did not survive to sexual maturity. However, males fed sugar and protein for<br />
only two days immediately after adult emergence or immediately before release did survive on<br />
water only and where more competitive sexually than males fed sugar only. Furthermore,<br />
females fed protein during two days did not mate.<br />
Conclusions: Providing protein supplements for a short period of time to females may not be<br />
enough for them to mature sexually, which could favor a unisexual release in this species.<br />
Protein supplements provided for only two days immediately after emergence or immediately<br />
before release may be enough to increase the sexual performance of sterile males without the<br />
detrimental effects on longevity.<br />
Keywords: Mexfly, protein diet, sexual competitiveness, longevity
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EXPOSURE OF STERILE MALES TO GINGER ROOT OIL REDUCES MATING IN WILD<br />
CERATITIS CAPITATA FEMALES (DIPTERA: TEPHRITIDAE).<br />
Morelli, Renata 1 ; Paranhos, Beatriz J* 2 ; Coelho, Mello A 3 ; Castro, Rosimary 2 ; Garziera,<br />
Luiza 2 ; Lopes, Fabiana 2 & Bento, Jose Mauricio S 1<br />
1 University of São Paulo, ESALQ, Department of Entomology and Acarology, C.P. 9, 13418-900 Piracicaba-SP,<br />
Brazil. E-mail: jmsbento@esalq.usp.br; 2 Embrapa Semi Arid, Laboratory of Entomology, Petrolina-PE, Brazil;<br />
3 University of São Paulo, ESALQ, Department of Exact Sciences, Piracicaba-SP, Brazil.<br />
Background: Females of Ceratitis capitata are facultatively polyandrous, with remating more<br />
common in laboratory strains rather than wild ones. In the application of the Sterile Insect<br />
Technique (SIT) against this pest, large overflooding ratios of sterile: wild males can increase<br />
the remating frequency, because females that mate for the first time with a sterile male tend to<br />
remate more frequently. The exposure of sterile males to ginger root oil (GRO) is used in C.<br />
capitata SIT programmes to increase the sterile male’s mating success. Exposing males to an<br />
aromatherapy with GRO results in greater competitiveness of sterile males when competing for<br />
wild females, and this may also increase the remating frequency among wild females.<br />
Methods: This work examined, under greatly male biased sex ratios, whether sterile male<br />
exposure to GRO has an effect on the remating behavior of wild females. The frequency of<br />
wild females remating, number of matings per female, the refractory period between the first<br />
and second mating, and the duration of the first and second matings of wild females were<br />
determined under laboratory conditions for three mating scenarios that included wild males<br />
only or wild males competing with sterile males (either GRO-treated or non-treated).<br />
Results: Wild females first mated with sterile males exposed to GRO had their remating rate<br />
over six days and the mean number of matings per female reduced in comparison to those first<br />
mated with non-exposed sterile males, from 62.5% to 31.1% and from 3.1 to 1.6, respectively.<br />
Conclusion: The remating parameters of females mated with sterile GRO-exposed males<br />
resembled those of females mated with wild males.<br />
Keywords: Ceratitis capitata, female receptivity, sterile insect technique, α-copaene<br />
229
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF METHYL EUGENOL EXPOSITION TO YOUNG STERILE MALES OF<br />
BACTROCERA DORSALIS(HENDEL) AND BACTROCERA CORRECTA (BEZZI) ON<br />
MATING SUCCESS.<br />
Pinkaew, Supaap*; Chinvinijkul, Suksom & Orankanok, Watchreeporn<br />
1 Department of Agricultural Extension, 2143/1 Paholyothin Road, Chatuchak, Bangkok 10900 Thailand. Email:<br />
pinkaew_mon555@yahoo.com<br />
Background: Area-wide integrated control of fruit flies are going on in Thailand while Areawide<br />
Sterile Insect Technique (SIT) program against fruit flies are increasingly implemented<br />
worldwide. Achievement of SIT is to assess mating success of released sterile males. Influence<br />
of methyl eugenol, ME, was generally known as effective to the sexual maturity male fruit<br />
flies. We reported here on effect of methyl eugenol on young sterile males as a precise marker<br />
to assess mating performance.<br />
Methods: Premature mass reared sterile males of the 32nd and 34th generations of two species<br />
tephritid fruit fly, (Diptera: Tephritidae), Bactrocera dorsalis (Hendel) and Bactrocera<br />
correcta (Bezzi), respectively were fed only water-agar after eclosion. The first group of sterile<br />
males, 2, 3, 4 and 5 day-old, were exposed to 1 ml ME from 8.00-9.00 a.m. The second group<br />
of the same ages were not exposed to ME and provided with standard adult diet, sugar protein<br />
hydrolysate, SP 3:1, and water al libitum. During 15 testing days of 50 B. dorsalis sterile males<br />
without ME, 50 sterile males with ME, and 50 mature wild males (23 day-old) were introduced<br />
into field cage competing for 50 of mature wild females (21 day-old). Bactrocera correcta<br />
were conducted and observed similar as B. dorsalis. But the testing day for B. correcta took 18<br />
days; mature wild males were 34 day-old; mature wild females were 37 day-old.<br />
Results: Bactrocera dorsalis sterile males, 2 day-old, exposed to ME showed significantly<br />
higher percentage of mated pairs than ones achieved by non methyl-eugenol and set of 3+n,<br />
4+n and 5+n day-old males after exposed 2-15 days. Higher percentage of mated pairs were<br />
shown on the first day after exposed than the group without ME and reached no significantly<br />
different on the 5th-8th and 10th, 13th-15th day after treated. Enhanced mating performance of<br />
older fly B. dorsalis strongly presented for 15 days. Meanwhile sterile males exposed to ME at<br />
age 3, 4 and 5 days were started no significant different from the 3rd, 2nd and 2nd day after<br />
treated, respectively. For B. correcta, percentage of mated pairs of all young sterile males<br />
exposed to ME at age 2, 3, 4 and 5 day-old were significantly higher than the ones achieved by<br />
non ME and reached non significant to wild males from the 6th-14th day. Older B. correcta<br />
flies treated at age 3, 4, 5 day-old showed high peak of the relative sterile index, RSI, that was<br />
non significant with wild males from the first day after treated. The relative sterile index of<br />
sterile males treated at age 3 and 4 days was better than wild males from the 8th-13th day and<br />
the 6th-11th day after treated, respectively.<br />
Conclusions: Young sterile males of B. dorsalis and B. correcta of 2, 3, 4 and 5 day-old<br />
exposed to ME showed manifest mating competitiveness significantly higher than non ME<br />
exposed steril males. But they were competitively inferior to mature wild males.<br />
Keywords: Bactrocera dorsalis, Bactrocera correcta, young sterile male, methyl eugenol expose, mating<br />
competitiveness
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
INFLUENCE OF GINGER ROOT OIL EXPOSURE AND POST-TENERAL NUTRITION ON<br />
LONGEVITY AND MORTALITY IN BAIT TREATMENTS OF STERILE MEDFLY.<br />
San Andrés, Victoria 1,2 , Pérez-Panadés, Jordi 3 ; Carbonell, Emilio 3 ; Castañera, Pedro 2 &<br />
Urbaneja, Alberto 1 *<br />
1 Instituto Valenciano de Investigaciones Agrarias (IVIA), Unidad Asociada de Entomología IVIA– CIB CSIC.<br />
Ctra. de Moncada a Náquera km. 4,5; Moncada 46113- Valencia (Spain). Email: aurbaneja@ivia.es. 2 Centro de<br />
Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC), Departamento<br />
Biología de Plantas, Madrid, Spain. 3 Instituto Valenciano de Investigaciones Agrarias - Biometría, Carretera<br />
Montcada-Nàquera Km 4.5, Moncada, Valencia 46113, Spain,<br />
Background:A key issue for Sterile Insect Technique (SIT) programmes against medfly,<br />
Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is to release sterile males that are<br />
sufficiently competitive with males from the wild population. Post-teneral nutrition and ginger<br />
oil (GRO) exposure of sterile males prior to release have been shown to improve male<br />
competitiveness or performance. However, few studies are available on the effect of postteneral<br />
nutrition and ginger oil exposure on longevity and mortality in bait treatments by sterile<br />
male C. capitata. Our study was based on the hypothesis that the increase of sexual activity of<br />
males exposed to GRO may have an energetic cost that induces them to search for food<br />
resources, such as proteins included in bait treatments.<br />
Methods: Four treatments were compared: (1) protein-fed flies not exposed to GRO, (2)<br />
protein-fed flies exposed to GRO, (3) protein-deprived flies not exposed to GRO, and (4)<br />
protein-deprived flies exposed to GRO. Longevity and mortality in bait treatments of sterile<br />
medfly of the Vienna 8 strain for the corresponding treatments were obtained.<br />
Results: Sterile male longevity was increased by the addition of protein to the standard prerelease<br />
sugar diet, whereas exposure to GRO did not influence their longevity. Mortality in<br />
spinosad baits was influenced both by diet and GRO exposure. Sterile males on a proteindeprived<br />
diet suffered greater mortality than sterile males fed with both sugar and protein.<br />
When sterile males were fed on the protein-deprived diet, GRO exposure increased their<br />
mortality. However, no significant differences were found in adults on the sugar-protein diet,<br />
whether or not they had been exposed to GRO.<br />
Conclusions: These results show, for the first time, a negative effect of GRO exposure in terms<br />
of increasing mortaliy in proteinaceous bait treatments, a common practice in areas where SIT<br />
is implemented. Nevertheless, this effect could be reduced by the addition of protein to the<br />
standard pre-release diet. The implications of these results for SIT programmes against C.<br />
capitata are discussed.<br />
Keywords: Sterile Insect Technique, SIT, medfly, sugar, protein, proteinaceous bait, Diptera, Tephritidae<br />
231
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PROTEIN RESOURCE ALTERNATIVES FOR MASS REARING OF CERATITIS CAPITATA<br />
WIEDEMANN (DIPTERA: TEPHRITIDAE).<br />
Silva Neto, Alberto M. 1 ; Santos, Tatiana R. O. 1 ; Dias, Vanessa S. 1 ; Joachim-Bravo, Iara<br />
S. 1 ; Benevides, Leandro J. 2 ; Benevides, Clicia M. J. 2 ; Silva, Mariangela V. L. 2 ; Santos,<br />
Deise C. C. 3 ; Virginio, Jair F.* 3 ; Oliveira, G. B. 4 ; Walder, Julio M. M. 5 ; Paranhos,<br />
Beatriz A. J. 6 & Nascimento, Antônio S. 7<br />
1 Universidade Federal da Bahia, Dep. de Biologia Geral, Campus Ondina, Av. Adhemar de Barros, Salvador-BA,<br />
Brazil; E-mail: bio.alberto@gmail.com; 2 UNEB- Departamento de Ciências da Vida- Campus 1, Salvador-BA,<br />
Brazil; 3 Biofábrica Moscamed Brasil, Juazeiro-BA, Brazil; 4 UFRB – Centro de Ciências da Saúde, Feira de<br />
Santana-BA, Brazil; 5 Centro de Energia Nuclear na Agricultura – CENA/USP, Piracicaba-SP, Brazil; 6 Embrapa<br />
Semiarido (CPATSA), Petrolina-PE, Brazil; 7 Embrapa Mandioca e Fruticultura Tropical (CPNMF), Cruz das<br />
Almas-BA, Brazil.<br />
Background: Ceratitis capitata is one of the most economic importance pest in fruit crops<br />
around the world and it’s control need to be a safety and environmental friendly technique,<br />
such as Sterile Insect Technique (SIT), that consists to mass rearing insects with high quality<br />
insect and low cost. Artificial diet is the major bottleneck in the Facility Moscamed Brazil<br />
(BMB), regarding the source protein (yeast hydrolysate) of the adult's diet, because it is<br />
expensive and has to be imported.<br />
Methods: Two commercial products, from national origin and low cost, were tested (yeast<br />
autolysed-AL and yeast extract-EL) as substitutes of imported protein. The biological<br />
parameters appraised were: fecundity, adults' longevity and eggs’ viability.<br />
Results: We founded that flies mass reared on the national protein sources were equivalent or<br />
superior to the flies on imported one, where females fed on EL protein in laboratory conditions<br />
presented higher fecundity and females fed on EL protein had higher longevity.<br />
Conclusions: Based in the obtained results the two commercial products demonstrated to be<br />
possible substitutes for imported protein (yeast hydrolysate) in C. capitata mass rearing, which<br />
can be easily found in the national market, and with low cost.<br />
Keywords: Mediterranean fruit fly, medfly, adult diet, facility, SIT.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
IMPROVED AND DEVELOPED OF ALTERNATIVE LARVAL DIETS FOR MASS<br />
REARING FRUIT FLIES SPECIES OF THE GENUS ANASTREPHA.<br />
Hernández, Emilio* 1 ; Rivera, Pedro 1 ; Bravo, Bigail 1 ; Salvador, Miguel 2 & Chang, Chiou<br />
L. 3<br />
1 Desarrollo de Métodos, Programa Moscafrut-Moscamed SAGARPA-IICA. Calle Central Poniente No. 14,<br />
Centro. 30700 Tapachula Chiapas, México. E-mail: emilioho@prodigy.net.mx; 2 Centro de Biociencias,<br />
Universidad Autónoma de Chiapas, Tapachula, Chiapas, México; 3 U.S. Pacific Basin Agricultural Research<br />
Center, USDA-ARS, 64 Nowelo St., Hilo, HI 96720.<br />
Background: Mass rearing fruit flies for Sterile Insect Technique programs needs artificial<br />
diets to maintain and developed larvae of the target species. The feasibility of any diet depends<br />
on the cost of the raw materials, their accessibility and quality. The Moscafrut Facility at<br />
Metapa, Chiapas, México has a continuous project for test available and developed new<br />
ingredients with potential to use for mass-production Anastrepha ludens and A. obliqua, and to<br />
maintain colonies of A. serpentina and A. striata with the goal to increase quality and decrease<br />
the cost.<br />
Methods: The work consisted of six experiments. In the first was determined the efficacy of<br />
new diets, jellified diet using corn, rice and potato starch (Nutrifly ® ), jellified diet using no<br />
purified agar (Agarmex) and liquid diet. In the second experiment were evaluated no typical<br />
protein sources, Tempeh (flour of chick pea fermented with Rhizopus oligosporus), Mubarqui ®<br />
(flour of Amaranthus seeds), lyophilized of fruits of Sargentia greggi and yeast extract;<br />
compared with Torula yeast and hydrolyzed protein. In the third experiment was evaluated the<br />
agar-started diet. In the fourth experiment was evaluated the elimination of the Methyl Paraben<br />
(Nipagen). In the fifth experiment was evaluated the Torula yeast content. In the last<br />
experiment were evaluated new commercial available torula yeast marks.<br />
Results: At experimental conditions the Nutrifly ® , liquid and agar diets increased the larval<br />
weight maintained the at least 70% of neonate larval survival for A. ludens, A. obliqua and A.<br />
serpentina. The Tempeh permitted high larval transformation and pupal weight in A. obliqua.<br />
The use of agar-started diet permitted the established of an experimental colony of A. striata.<br />
The absence of Nipagen had not any effect of the production and quality parameters. A. ludens,<br />
A. obliqua and A. serpentina required diets with 9, 7 and 5% content, respectively. Diets<br />
prepared with Lake States ® , Nutribio ® or Lallemand ® yeast produced excellent results in<br />
experimental tests for A. ludens and A. obliqua.<br />
Conclusions: Jellified diet with starch and agar have high potential to use as started diet, which<br />
as liquid diet permit save space and production cost. Tempeh, Mubarqui ® , and lyophilized of<br />
fruits need to improve but today they could be used to substitute partially the torula yeast.<br />
Started diet permits high larval survival during the colonization and establishment of new<br />
colonies. Nipagen is not necessary for larval survival in the diet. Lake States ® , Nutribio ®<br />
(Biosaf) and Lallemand ® yeast permitted survival larvae. Protein sources as distiller´s dried<br />
grains and soluble (DDGS) should be tested.<br />
Keywords: Anastrepha, larval diets, protein sources, started diets, torula yeast<br />
233
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF ENSILED RICE STRAW AND FRUIT FLY LEFTOVERS ON BEEF CATTLE<br />
PERFORMANCE.<br />
Orankanok, Somchai* 1 ; Thumdee, Patama 1 ; Katepongpun, Wichit 1 & Orankanok,<br />
Watchreeporn 2*<br />
1<br />
Department of Animal Sciences, Faculty of Agriculture and Natural Resources, Rajamangala University of<br />
Technology Tawan-Ok (RMUTTO), Chonburi. 20110. Thailand. . Email: orankanoks@hotmail.com; 2<br />
Department of Agriculture Extension, Ministry of Agriculture and Cooperatives; 2143/1 Phahol Yotin Road,<br />
Chatuchak, Bangkok, 10900. Thailand.<br />
Background: Fruit fly mass rearing need a large amount of larva diet which is wheat bran based<br />
diet. Spent larva diet or fruit fly leftovers (FFL) still contain nutritive value for livestock as<br />
shown by proximate analysis of 57.98 % moisture, 12.7 % crude protein, 1.86 % fat, 19.36 %<br />
fiber, 7.84 % ash, 0.05 % calcium and 0.2 % phosphorous. Application of fruit fly leftovers as<br />
livestock feed could save cost of environmental problem in term of disposing the used diet and<br />
save feed cost of livestock production.<br />
Methods: Change over design was conducted to evaluate the effectiveness of 4 treatments. A;<br />
ensiled 95 kg. of 4 % urea-rice straw + 5 kg. of malasses (control). B; ensiled 60 kg. of 5 %<br />
urea-rice straw + 5 kg. of malasses + 35 kg. of FFL. C; ensiled 45 kg. of 5 % urea-rice straw +<br />
5 kg. of malasses + 50 kg. of FFL. D; ensiled 30 kg. of 5 % urea-rice straw + 5 kg. of malasses<br />
+ 65 kg. of FFL. 16 hybrid cattle were assigned as individual effect and 4 phases of feeding<br />
were assigned as row effect. Each phase of feeding was 4 weeks with 2 weeks of resting<br />
period.<br />
Results: Four treatments of cattle feed show gradually decreased from 8.5 (A; control), to 7 (B;<br />
35 % FFL), 6.5 (C; 50 % FFL), and 5.5 (D; 65% FFL) corresponding with increasing of strong<br />
smell of ammonia. The first 3 groups of ensiled feed were yellow-to-brown in color whereas<br />
feed D produced brown-to-dark brown color. Average daily gain (kg./day) of feed A, B, C and<br />
D were 0.790, 0.710, 0.840 and 0.620, respectively and found significant difference (p
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
PERFORMANCE OF GROWING SWINE FED BY DIETS SUPPLEMENTED WITH<br />
DIFFERENT LEVELS OF FRUIT FLY MASS-REARING DIET LEFTOVERS.<br />
Orankanok, Somchai* 1 ; Watechakul, Narumon 1 ; Chinvinijkul, Suksom 2 & Orankanok,<br />
Watchreeporn 2 *<br />
1 Department of Animal Sciences, Faculty of Agriculture and Natural Resources, Rajamangala University of<br />
Technology Tawan-Ok (RMUTTO), Chonburi. 20110. Thailand. Email: orankanoks@hotmail.com; 2 Department<br />
of Agriculture Extension, Ministry of Agriculture and Cooperatives, 2143/1 Phahol Yotin Road, Chatuchak,<br />
Bangkok, 10900, Thailand. Email: Watchreporn@yahoo.com<br />
Background: Larva diet, wheat bran based diet, in process of fruit fly mass rearing consist of<br />
wheat bran, granulated sugar, instant dry yeast, sodium benzoate, methyl-p-hydroxybenzonate,<br />
acetic acid and water. Nutrients in the used larva diet or fruitfly leftovers(FFL) were shown by<br />
proximate analysis of 57.98 % moisture, 12.7 % crude protein, 1.86 % fat, 19.36 % fiber, 7.84<br />
% ash, 0.05 % calcium and 0.2 % phosphorous with amino acids frofile of 0.44 % alanine, 0.31<br />
% arginine, 0.13% cystine, 0.37 % glycine, 0.32 % histidine, 0.22 % isoleucine, 0.56 %<br />
leucine, 0.23 % lysine, 0.13 % methionine, 0.31% phenylalanine, 0.27 % threonine, 0.13 %<br />
tryptophan, 0.18 % tyrosine and 0.31 % valine. The mentioned nutrients are benefit to produce<br />
swine feed, especially feed for growing- finishing swines since major cost of production of this<br />
period is feed cost.<br />
Methods: Randomized complete block design was applied to evaluate experimental feed. Growing<br />
swines, 60 kg. of body weight, were divided in to 4 groups. Each group was composed of 8 swines.<br />
Treatments were grower diet varied by different levels of dried fruit fly leftovers as followed :<br />
treatment 1, grower diet with 20 % rice bran without FFL (control); treatment 2,<br />
grower diet with 7.5 % rice bran + 7.5 % FFl; treatment 3, grower diet with 0 % rice bran + % 15<br />
FFL; treatment 4, grower diet with 0 % rice bran + 20 % FFL. Each diet group was fortified by 3,200<br />
kg cal/kg ME, 18 % protein and other nutrient requirements followed by NRC(1998). All of swines<br />
were fed by full feeding until about 100 kg. of body weight.<br />
Results: Palatability of diets mixed with FFL were a bit better than control group resulting in higher<br />
feed intake and faster growth rate in term of average daily gain which were 0.872, 0.967, 0.970 and<br />
0.851 gram/day for control, 7.5 % FFl, 15 % FFL and 20 %FFL, respectively. Growth rate of 7.5 and<br />
15 % FFL were significantly higher than 20 % FFL (p
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SEXUAL MATURATION IN A MUTANT STRAIN OF ANASTREPHA FRATERCULUS<br />
(DIPTERA: TEPHRITIDAE).<br />
Peralta, Patricia A.; Segura, Diego F.*; Milla, Fabián H. & Cladera, Jorge L.<br />
Instituto Nacional de Tecnología Agropecuaria (INTA), Instituto de Genética "Ewald Favret" (IGEAF) De los<br />
Reseros y Repetto (1712) Castelar, Buenos Aires, Argentina. pperalta@cnia.inta.gov.ar<br />
Background: Sexual maturation is a relatively long process in Anastrepha males. This leads to<br />
an increase in the costs to apply the sterile insect technique (SIT) because the flies must be kept<br />
in laboratory conditions for several days before released. Our objective was to study the<br />
duration of the sexual maturation process in a mutant strain in Anastrepha fraterculus and to<br />
study the heretability of this trait.<br />
Methods: To estimate sexual maturity we observed the sexual behavior of males at different<br />
ages, after offering them sexually mature virgin females. The percentage of males that mated at<br />
different ages was the variable under study. We analyzed two strains: an eye color mutant<br />
(#3210) and a wild-type strain as control (L-TUC). We found that males from strain #3210 had<br />
a significantly shorter pre-copulatory period than the males from the other strain. To study the<br />
heredability of this character in the mutant strain, we performed crosses between individuals<br />
from the #3210 strain and the males and females of wild-type stock.<br />
Results: We observed that the sons of #3210 males matured significantly faster than the<br />
descendants of wild-type males, irrespective of the origin of the mother. When the descendants<br />
of both sexes were analyzed at the 6 th and 13 th generation, we found that these results were<br />
repeated.<br />
Conclusions: These results encourage the use of this strain of A. fraterculus both as model to<br />
study the inheritance of genes associated to the sexual maturation, and also as a candidate<br />
strain for massive rearing and release with the framework of an SIT program against this fruit<br />
fly pest.<br />
Keywords: sexual maturation, Anastrepha fraterculus mutant, males pre-copulatory period.
Session 6<br />
Area-Wide & Action Programs
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
SIT PILOT PROJECT IN CROATIA; CONTROL OF MEDFLY BY SIT IN THE NERETVA<br />
VALLEY<br />
Bjelis, Mario¹; Marusic, Ivana¹ & Popovic, Luka²<br />
¹Croatian Center for Agriculture, Food and Rural Affairs, Zagreb; Zvonimirova 14 a, 21210 Solin,<br />
mario.bjelis@hcphs.hr; ² Croatian Center for Agriculture, Food and Rural Affairs, SIT Emergency facility, Tisno<br />
bb, 20355 Opuzen.<br />
Background: The Neretva river valley is divided with a part in the Republic of Croatia and a<br />
part in Bosnia and Herzegovina. It has been noted that highest damage caused by medfly –<br />
Ceratitis capitata Wied. (Diptera, Tephritidae) is along the border area and can cause up to<br />
30% damage on mandarines. The Croatian Government strongly supports the project since the<br />
medfly became a serious pest in the valley. Interest from the Croatian Government to integrate<br />
the sterile insect technique (SIT) on an area-wide basis as an alternative for insecticide-based<br />
medfly control in consultation with the Joint FAO/IAEA Division determined that Neretva<br />
river valley could be a good location to apply this environmentally friendly pest control method<br />
as part of an integrated pest management system. The Neretva valley, is the largest citrus<br />
production area in Croatia.<br />
The Neretva river valley is a relatively isolated ecosystem (similar to that of the Hex River<br />
Valley in South Africa where IAEA had supported activities of medfly control by using of<br />
SIT). This provides a good probability of success for a future project since such relative<br />
isolation by mountains would limit the movement of wild medfly adults into the valley. A<br />
national project involving the Neretva Valley as a pilot project for southern Europe is feasible<br />
in view that it represents an ecosystem at the northern edge of medfly distribution.<br />
Methods: An pilot area of 650 ha of mandarine was selected as pilot area where sterile males<br />
are released since april 2010. The files were produced in BioFly Israel, and were shipped<br />
directly to Croatian airport Split, twice per week. A total of 70 shipments will be delivered to<br />
Croatia during 2010, as a part of FAO/IAEA RER 5014 TCP.<br />
Croatian government build an emergency facility with capacity of 20 mil flies/week in city of<br />
Opuzen in Neretva river valley.<br />
Results and Conclusions: The Neretva valley includes over 80% of the Croatian national citrus<br />
production, valued at USD 13 million of exported mandarine fruit per year, mostly to EU. The<br />
medfly control by integrating SIT technique would ecconomically benefit all local stakeholders<br />
including growers and the public at large in view more access to fresh fruits. This program will<br />
benefit from the support of the local authorities as well as of the growers and exporters.<br />
Keywords: Neretva river valley, SIT technique, pilot project<br />
239
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE CUBANS EXPERIENCES ON MONITORING AND MANAGEMENT OF FRUIT FLY<br />
ANASTREPHA SPP (DIPTERA:TEPHRITIDAE) IN MANGO (MANGIFERA INDICA L)<br />
AND GUAVA (PSIDIUM GUAJAVA. L ) CULTURE.<br />
Borges, Mirtha*; Beltrán, Alina; Mulkay, Tania; Rodriguez, Jorge L.; Hernández, Doris<br />
& Paumier, Adrian .<br />
Dirección de Ciencia y Técnica, Grupo de Ecología y Manejo de Plagas, Instituto de Investigaciones en<br />
Fruticultura Tropical (IIFT), Miramar Playa, Ciudad de la Habana, Cuba; Email: mirthaborges@iift.cu.<br />
Background: Agricultural programs for sustained fruit tree production have increased during<br />
last years, contributing to availability of products for consumption and marketing. Tephritid<br />
fruit flies constitute one of the main pests of tropical and subtropical fruit trees. The presence<br />
of fruit fly species of Anastrepha genus in fruit tree culture has originated the application of<br />
quarantine regulations and restrictions of importing and exporting countries Of them, due to<br />
importance for Cuban fruitculture stand out seven, being of the greater impact: "Caribbean fruit<br />
fly", Anastrepha suspensa Loew, "West Indian fruit fly" Anastrepha obliqua Mcquart, in guava<br />
and mango indistinctly.<br />
Methods: Studies on mango and guava cultures were done with the following objectives:<br />
taxonomic identification, monitoring systems (traps, attractants and fruit sampling, occurrence<br />
of parasites), population behavior, and prospecting and control methods. Entomological<br />
inspections were done to detect the presence of fruit fly parasitoids in traps with attractants and<br />
collected fruits. The use of chemical, agrotechnical, cultural biological and postharvest control<br />
methods were tested in mango and guava cultures.<br />
Results: The obtained result was that Anastrepha obliqua and A. suspensa were the species of<br />
more frequency in the culture. It is proposed as result the of use Torula yeast as attractant<br />
alternative with varieties and contributions to monitoring and sampling system, alternative host<br />
plant ,population behavior ,biological control (prospection & application ) with the detection<br />
by first time in Cuba the larvae parasite Utetes anastrephae ( Hymenoptera: Braconidae ) and<br />
the application of the entomopathogenous nematodes Heterorabhditis indica on larvae and<br />
pupae of these species of fruit flies. The agrotechnical, cultural and chemical (insecticide- bait)<br />
methods showed good results. The effectivity of treatment postharvest with the use of hot<br />
water on the fruits of mango cv Super Haden, presented 4 for the egg stadium and zero for<br />
larvae, while with wax cover, a total of 48 and 53 alive larvae were obtained. In the<br />
combinations wax cover + hot water, the control of larvae was of 100%. This effectivity of<br />
control pre & postharvest are important and insert in the IPM Program.<br />
Conclusions: The use of alternative attractants, the knowledge of the behavior, hosts plants of<br />
Anastrepha spp, the agrotechnical, chemical cultural, biological control and postharvest assays<br />
offer promissory result in this work line of importance for the economic and quarantine point<br />
of view. These results constitute the principal basis of IPM on fruit flies of economic<br />
importance in Cuba conditions.<br />
Keywords: Anastrepha, monitoring, mango guava, pre and postharvest control
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
ADVANCES IN THE NATIONAL PROGRAMME AGAINST FRUIT FLIES IN MÉXICO.<br />
Gutiérrez-Ruelas, José M.*; Santiago-Martínez, Guillermo; Cárdenas-Lozano, Jesús &<br />
de los Santos, Martín.<br />
Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria (SENASICA). Dirección General de Sanidad<br />
Vegetal (DGSV). Av. Guillermo Pérez Valenzuela No. 127 Colonia del Carmen Coyoacán, C.P. 04100 México,<br />
D. F. Email: jruelas@senasica.gob.mx<br />
Background: In 1992 the Government of Mexico established the National Program against<br />
Fruit Flies (NPFF) in order to control populations of Anastrepha ludens, A. obliqua, A. striata<br />
and A. serpentina, and establish Free Areas of Fruit Flies (FA-FF) and Areas of Low<br />
Prevalence of Fruit Flies (ALP-FF), under an Area-Wide Integrated Pest Management (AW-<br />
IPM) approach, including the Sterile Insect Technique (SIT) and the Augmentative Biological<br />
Control (ABC). In this paper we report the most significant achievements in the period 2001 -<br />
2010.<br />
Methods: We established strategic alliances among the Federal Government, State<br />
Governments and fruit producers, to combine efforts and economic resources to implement the<br />
Strategic Plan for the control of the pest. We signed an Agreement Cooperation with the<br />
Interamerican Institute for Cooperation on Agriculture (IICA), for the production of sterile flies<br />
A. ludens and A. obliqua, and the parasitoid Diachasmimorpha longicaudata. With the Institute<br />
of Ecology AC, and the International Atomic of Energy Agency (IAEA) were implemented<br />
specific projects to perform basic and applied research. Currently, the SIT is applied on 100<br />
000 ha of citrus, mango and guava, for which we operate five centres of Packaging and Release<br />
of sterile flies under the system of chilling adult and aerial releases. On a weekly basis, in this<br />
facilities are packed an average of 215 million of sterile pupa, using PARC boxes or towers<br />
type “Mexico” or “Worly”. After emergence, the adult flies are supplied with juvenile hormone<br />
through the food, in order to accelerate sexual maturity.<br />
Results. In 2009 we produce 9.500 million of A. ludens sterile, 2.300 million of A. obliqua<br />
sterile and 1,600 million of D. longicaudata. Until May 2010, 49.91% (978.093 km2) of the<br />
national territory has been recognized as free area of fruit flies (FAFF), and 10.7% (210.603<br />
Km2) as low prevalence areas of fruit flies (ALP-FF). From free areas, we have been exported<br />
160 000 tons of mango, 120 000 tons of orange, and 7.300 tons of peach, with a trade value of<br />
U.S. $ 190 million. In our area of Methods Development, we have more strains of parasitoids<br />
(e.g., Coptera haywardi and Doryctobracon crawfordi), which in short time could be scaled to<br />
mass production with the aim to strengthen the biological control of the pest. In our project of<br />
genetic sexing, we obtained a new strain of A. ludens which is differentiated by black pupae in<br />
females, which allows us the exclusive selection of males. This condition is a relevant attribute<br />
for the SIT, so we have started evaluating the quality of the strain, and scaling it for mass<br />
production.<br />
Conclusions. The economic evaluation of this Programme in six states of the Mexican<br />
Republic during the period of 1994-2008, demonstrates the benefits of the implemented<br />
actions, which reflects that investment has a positive cost-benefit ratio.<br />
Keywords: Area-Wide, IPM, SIT, Anastrepha spp. Pest Free Areas.<br />
241
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
SMALL ACREAGE AREA-WIDE CONTROL OF BACTROCERA DORSALIS HENDEL ON<br />
CITRUS.<br />
Huang, Yu-Bin 1,2* ; Kao, Ching-Hua 1 ; Chiang, Ming-Yao 1 & Cheng, Edward Y. 1<br />
1 Applied Zoology Division, Taiwan Agricultural Research Institute, Wufeng, Taichung 41362, Taiwan, ROC, Email:<br />
ybhuang@tari.gov.tw; 2 Department of Entomology, National Chung Hsing University, Taichung, 40998<br />
Taiwan, ROC.<br />
Background: Citrus is a very important fruit crop in Taiwan with an acreage of more than<br />
32,000 ha. The oriental fruit fly (Bactrocera dorsalis Hendel, OFF) is a constant problem on<br />
the crop, and since citrus is such an important export commodity, the OFF control program has<br />
been on the priority list ever since 1959. Following the success of area-wide control of OFF in<br />
sugar apple (in Taitung) and wax apple (in Juan-Nan), similar studies to control OFF damage<br />
on citrus were deemed appropriate.<br />
Methods: Da-Lin Township of Chia-Yi County was selected for this study, because citrus is the<br />
only fruit crop grown in this area. With the cooperation of area growers, an area-wide OFF<br />
control program was launched in 2005. It included a four step process: (1) Growers were<br />
organized into a cooperative control team. (2) OFF population densities were monitored, and<br />
appropriate geographic and agricultural data were collected. (3) Beginning two months prior to<br />
and lasting until the end of harvesting, methyl eugenol bait was used to attract and kill OFF. (4)<br />
Field sanitation. The control program was carried out in a 400 ha test area which included 120<br />
ha citrus orchards (60,000 trees) beginning in early June, 2005. No buffer zone was set as<br />
protect orchard and spatial warming.<br />
Results: The citrus production in Da-Lin was not subjected to bagging as was done in the wax<br />
apple and sugar apple studies. As such, the OFF damage rate was assessed directly. The OFF<br />
population was reduced to less than one fly per trap per day in comparison to historical data.<br />
By sampling the citrus fruits from three orchards, a fruit damage rate of 0.35% was observed.<br />
An additional benefit of this program was that no insecticides were needed. Reducing the OFF<br />
damage minimized not only the insecticide cost, but eliminated the insecticide residue problem<br />
as well. The quality of the citrus fruits was also improved. For each of the two consecutive<br />
years, eliminating the use of insecticide spray resulted in a cost saving estimated at 1.4 million<br />
NT dollars (US$ 44,343)/ year .<br />
Conclusions: Economic motivation and grower education are critical incentives in a successful<br />
area-wide control program. The success of such small acreage control programs can be used as<br />
a model for extention into other similar production areas in Taiwan in future studies.<br />
Keywords: Area-wide control, Bactrocera dorsalis, Citrus.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
MANAGEMENT OF FRUIT FLY, BACTROCERA CUCURBITAE (COQUILLETT)<br />
INFESTING GHERKINS USING AREA WIDE CONTROL.<br />
Hunasaghatta, Praveen M. 1 ; Muniyappa, Nandeesh 1 ; Chandra Mouli, Munikote R. 1* ;<br />
Gollahalli, Gundu R. V. 1 & Shanmugam, Vijaysegaran 2<br />
1 Global Green Company Limited, #14, 80 feet road, 4 th Block, Koramangala, Bangalore – 560 034, India. Email:<br />
chandramouli.m@globalgreencompany.com; 2 Entomologist Emeritus, Brisbane, Australia.<br />
Background: The melon fly, Bactrocera cucurbitae, is a serious pest of gherkin (Cucumis<br />
sativus), which is cultivated by small individual farmers but collectively on a large scale<br />
through contract farming in southern peninsular India for export to world markets. Intensive<br />
control measures undertaken by individual gherkin farmers were insufficient as fruit damage<br />
was above 4%, and highly unacceptable to the export market. An area wide management<br />
(AWM) program using a combination of male annihilation technique (MAT), protein bait<br />
application technique (BAT) and sugar baits was thus attempted for more effective<br />
management of the melon fly. We report here on the success of this area wide management<br />
program in suppressing melon fly populations, and reducing and maintaining melon fly<br />
damage on gherkin at levels acceptable for export markets.<br />
Methods: The area wide management (AWM) program was undertaken over an area of<br />
approximately three square km in Kashapura village of Gauribidanur taluk, Chickaballapura<br />
district, southern Karnataka, over the period April 2008 to March 2010. The AWM area<br />
comprised gherkin (10%), other crops (50%) and fallow area (40%) within a central zone<br />
surrounded by a 50m buffer zone. The control methods used were male annihilation technique<br />
(MAT), protein bait application technique (BAT), and sugar baits. Male annihilation was<br />
achieved through cue lure + chlorpyriphos blocks (Nomate life time* from Agri Land) placed<br />
at 50 m intervals within the central zone, and 25 m apart within the buffer zone. The cue lure<br />
blocks were replaced monthly. Protein bait (Prima protein bait from Malaysia) plus malathion<br />
spot sprays were applied at weekly intervals to the gherkin crop. In addition, a sugar (10 %<br />
jaggery) plus malathion solution was applied to the maize grown as a border crop surrounding<br />
gherkin. The same treatments were imposed in individual gherkin plots situated 3 km away<br />
from the AWM area. To evaluate the effectiveness of the AWM program, melon fly<br />
populations (number of fruit flies trapped per day - FTD) were monitored continuously using<br />
cue lure traps that were cleared weekly. Levels of fruit fly damage to gherkin fruits at harvest<br />
were assessed by dissection of fruits in the factory..<br />
Results: Treatments with MAT, BAT and sugar bait sprays in the AWM area trapped 0.9 fruit<br />
flies per day during first week of January 2008 and this was reduced to zero during the last<br />
week of April 2010. In comparison, treatments with MAT, BAT and sugar bait sprays in the<br />
control area outside the AWM area still trapped 0.02 flies in the last week of April 2010. In<br />
harvested gherkin fruits, fruit fly infestation in the AWM area was reduced to 0.3 %, which is<br />
acceptable by the factory compared to an infestation level of 1.5% in the control area.<br />
Conclusions: Area wide management with treatments, MAT, BAT and sugar bait sprays were<br />
highly effective in reducing infestation by melon fly in gherkin to less than 0.3 per cent, and<br />
enabled the gherkin industry in southern India to meet with the stringent quality required by<br />
international export markets.<br />
Keywords: Bactrocera cucurbitae, MAT, BAT, Area wide management, FTD.<br />
ESTABLISHMENT OF A COLONY OF THE NEW WORLD SCREWWORM,<br />
COCHLIOMYIA HOMINIVORAX, FOR COMING AW-IPM PROJECTS IN<br />
243
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
MERCOSUL.<br />
Mastrangelo, Thiago* 1 ; Fernandes, Thiago 1 ; Azeredo-Espin, Ana M. L. 2 ; Malavasi, Aldo 3<br />
& Walder, Julio M.M. 1<br />
1 Center for Nuclear Energy in Agriculture (CENA/USP). Av. Centenário 303, 13400-970, Piracicaba, SP, Brazil.<br />
Email: piaui@cena.usp.br; 2 Centro de Biologia Molecular e Engenharia Genética (CBMEG) (DGEB/IB),<br />
Universidade Estadual de Campinas (UNICAMP). P.O. Box 6010, 13083-875, Campinas, SP, Brazil; 3 Biofactory<br />
MOSCAMED Brazil (BMB). Distrito Industrial do São Francisco, 48900-000, Juazeiro, Bahia, Brazil.<br />
Background: Since the 1950’s, the AW-IPM programs integrating the SIT against the New<br />
World Screwworm (NWS), Cochliomyia hominivorax (Diptera: Calliphoridae), have been<br />
successful. The NWS was eradicated from the USA, Central America to Panama, but in most<br />
tropical regions of Latin America (excluding Chile), the NWS is still a serious threat to<br />
livestock. There is a raising interest for its suppression in South America. Between January and<br />
May 2009, a pilot-project, supported by the BID and COMEXA, was performed at the Brazil-<br />
Uruguay border. As the results were positive, novel regional AW-IPM projects can be planned.<br />
To set a mass-rearing center based in South America is strategic when considering long-term<br />
programs. The CENA/USP has a pilot-facility, built by the Agriculture Ministry and supported<br />
by FAO/IAEA, which serves to give technical support to SIT projects adopted by the<br />
government. In partnership with the Biofactory MOSCAMED Brazil, a project to produce<br />
sterile NWS started on 2009.<br />
Methods: The project aims to maintain a colony of a regional NWS strain, to develop a massrearing<br />
system and a sterilization protocol by X rays, and to study the sterility induction in<br />
regional strains. So far, a colony has been successfully established. The parental generation<br />
came from UNICAMP pupae (originally from Goiás State, Central Brazil). The adults are kept<br />
in cages (90x145x75 cm) and fed on a diet (honey, spray dried blood and brewer’s yeast-<br />
Bionis ® ). The oviposition substrate is a mix of larval diet disposal and blood. The larvae grow<br />
in a medium made of spray dried blood, spray dried egg, milk, sodium citrate, citric acid,<br />
water, formalin and ground meat (7%: 2%: 1%: 0.1%: 0.1%: 100%: 0.1%: 70% (w/v)). On the<br />
5 th day, larvae pupate in vermiculite. The colony is maintained at controlled rooms (38±1ºC;<br />
60±10% RH).<br />
Results: The colony has been reared for 7 generations. Egg hatch has been of 70±10%. From<br />
G1 to G7, the total amount of pupae produced was about 8 L (~ 67,000 pupae). The mean adult<br />
emergence and sex ratio (♀/♂+♀) were 86.7±3% and 0.59±0.08 respectively. Currently, only<br />
1.5 L pupae/generation is produced (cost ~ US$ 35.00), what is enough to conduct the rearing<br />
and sterilization bioassays. Tests to eliminate the meat from the larval diet and to introduce<br />
local bulk agents (as sugarcane bagasse and coconut fiber) are underway and will decrease the<br />
rearing cost.<br />
Conclusion: This NWS colony will be soon transferred to the BMB to attend coming SIT<br />
projects in MERCOSUL.<br />
Keywords: SIT, Screwworm, Cochliomyia hominivorax.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
AREA-WIDE BACTROCERA CUCURBITAE (COQUILLETT) CONTROL IN SELECTED<br />
AREAS OF MAURITIUS.<br />
Sookar, Preeaduth* 1 ; Hendrichs, Jorge 2 ; Alleck, Malini 1 ; Buldawoo, Indrani 1 ;<br />
Khayrattee, Fazilla B. 1 ; Choolun, Tejanand 1 ; Permalloo, Shradhanand 1 & Rambhunjun,<br />
Mahen 1<br />
1 Entomology Division, Agricultural Services, Ministry of Agro Industry Food Production & Security, Reduit,<br />
Mauritius. Email: psookar@mail.gov.mu. 2 Insect Pest Control Section, Joint FAO/IAEA Division, PO Box 100,<br />
Wagramer Strasse 5, A-1400 Vienna, Austria<br />
Background: The melon fly, Bactrocera cucurbitae (Coquillett), is the most important insect<br />
pest of cucurbit crops in Mauritius (Mascarenes island in the Indian Ocean) causing heavy<br />
damage. To control this pest, planters have recourse to regular conventional insecticide cover<br />
sprays with all their disadvantages. Besides, this practice adds up to their cost of production. A<br />
project entitled Feasibility Study for the Suppression of the Melon Fly in Selected Areas of<br />
Mauritius – MAR 5/016 was launched in June 2007. The main objectives of the project are to<br />
produce quality cucurbits through effective environment-friendly suppression methods and to<br />
minimise the use of pesticides for melon fly management.<br />
Methods: An isolated area of 110 ha occupied by some 135 cucurbit growers was selected. The<br />
growers were sensitised on the project through meetings, pamphlets, stickers and video film.<br />
As an incentive to participate fully in the project, they were all given sprayers, fruit disposal<br />
cages, protein bait and MAT blocks. The growers were introduced to the techniques used in<br />
area-wide melon fly suppression. Gradually they started to adopt the recommended methods,<br />
which included targeted bait applications, male annihilation technique and sanitation. Fruit<br />
flies were monitored through dry traps baited with Cuelure and Malathion, wet traps baited<br />
with modified waste brewer’s yeast and fruit collection and incubation in the labioratory.<br />
Results: Cucurbit infestation which was above 30% before project implementation was reduced<br />
to 5% one year later. A decrease in the frequency of pesticide application was noted. 25% of<br />
the planters were treating their crops thrice a week before the project was started compared to<br />
5% one year after project implementation. An increase in cucurbit production was noted. 85%<br />
of planters claimed to have obtained an increase in both quality and quantity of cucurbits. The<br />
melon fly population was reduced by 50%.<br />
Conclusions: Cucurbit growers were able to produce quality cucurbits by adopting<br />
environment-friendly techniques of melon fly control. The cost of cucurbit production was<br />
reduced through a decrease in use of pesticides as well as application costs. The project has<br />
demonstrated the use of an integrated approach to reduce insecticide use, and to produce better<br />
quality fruits.<br />
Keywords: Melon fly, trapping, bait application, male annihilation technique<br />
245
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
NEW ACHEIVEMENT OF FRUIT FLY SUPPRESSION PROGRAM IN THAILAND: CASE<br />
STUDY IN SAMUT SAKORN PROVINCE.<br />
Telavanich, Thitipong *1 ; Amornsak, Weerawan 2 & Orankanok, Watchreeporn 3<br />
1 Thai Fruit and Vegetable Producer Association, 29/10 Soi Charoennakorn 12, Charoennakorn Road, Klongbonsri,<br />
Klongsan, Bangkok. 10600. Email: okra@qualitygreen.com; 2 Department of Entomology, Faculty of Agriculture,<br />
Kasetsart University, Bangkhen Campus, Chatuchak, Bangkok, 10900, Thailand; 3 Department of Agriculture<br />
Extension, Ministry of Agriculture and Cooperatives, 2143/1 Phahol Yotin Road, Chatuchak, Bangkok, 10900.<br />
Thailand.<br />
Background: Fruit fly control has been focused for more than 20 years in Thailand using the<br />
Sterile Insect Technique-SIT, along with other safe methods. The successful technologies<br />
conducted in pilot areas have been conveyed and modified to be suitable in each other areas.<br />
Base on knowledge of area-wide fruit fly integrated control, Samut Sakorn province was<br />
proposed, which is traditional scatter fruit orchards over more than 100 km 2 . Various fruits<br />
grown in this province are major hosts of tephritid fruit fly (Diptera: Tephritidae). Fruit<br />
production in this area is presented all year round causing of fruit fly outbreak and fruit damage<br />
which might be very high up to 100 percent if there is no any insecticide applied. Sprays and<br />
young fruit bagging are normally used but costly to control fruit fly infestation. Samut Sakorn<br />
Provincial Administration Organization has embarked and supported the growers for<br />
controlling Tephritid fruit fly by collaboration with the Department of Agricultural Extension,<br />
Ministry of Agriculture and Cooperatives.<br />
Methods: To decrease fruit fly population, the committee of Samut Sakorn Provincial<br />
Administration Organization suggested that various aspects, outbreak problems by growers,<br />
growers’s attitude on fruit fly control methods and etc., should be considered and then growers<br />
would be educated on cause of the outbreak, suitable and effective fruit fly control and growers<br />
participation. To increase empowerment of growers, at the beginning of activities, growers were<br />
participated activities together, taught to make traps using polyethelene terephthalate-PET with<br />
methyl eugenol as a fly attractant. Growers were motivated by given fertilizer, value 1,000 Baht/<br />
2 litre of flies, as award if they could show captured fruit flies from their own handmade traps.<br />
Results: Growers in Samut Sakorn province were interested in all activities organized by the<br />
committee. They gained the knowledge, continue to cooperate and also help each other in<br />
controlling fruit flies. However, main achievements are the changed of growers’ attitude on<br />
mass trapping using methyl eugenol, decreasing of fruit fly population, lower fruit infestation<br />
and better visible quality.<br />
Conclusions: Campaign of area-wide fruit fly mass trapping could be simply implemented by<br />
growers and local government cooperation. Convincing the growers to use fruit fly traps and<br />
put more effort following neighbour success would be the way to stimulate them. Samut<br />
Sakorn Provincial Administration Organization has made a plan to establish fruit fly<br />
surveillance using trapping network and implementation of other appropriate environment<br />
friendly technologies.<br />
Keywords: fruit fly suppression, Samutsakorn province, mass trapping, area-wide, methyl eugenol.
Session 7<br />
Natural Enemies & Biocontrol
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EVALUATION OF ENTOMOPATHOGENIC FUNGI TO OF CERATITIS CAPITATA.<br />
Ali, Ali*; Sermann, Helga & Büttner, Carmen<br />
Faculty for Agriculture and Horticulture. Department for Crop and Animal Sciences. Division Phytomedicine.<br />
Humboldt university zu Berlin. Germany. Email aligermany80@yahoo.de.<br />
Background: Laboratory experiments were carried out to measure the pathogenicity and to<br />
study the infection process of isolate 412 of Beauveria bassiana, and isolate V24 of<br />
Lecanicillium muscarium to old larvae of the Mediterranean fruit fly, Ceratitis capitata<br />
(Diptera: Tephritidae) under the laboratory conditions after inoculation of the fungal<br />
suspension on the soil surface.<br />
Methods: To evaluate the potential of the fungi to old larvae, plastic doses (3.8 cm × 2.8 cm)<br />
were filled with dry soil and sprayed with 3 ml suspension (1 × 10 8 Spores/ml) on the soil<br />
surface using a small dash bottle. The density of spores valued (2.6 × 10 7 Spores/cm 2 ) on the<br />
soil. After soil application the old larvae were put on the treated soil. The plastic doses were<br />
then incubated at 25°C in the dark. Nine days after the treatment, the number of pupae,<br />
moulded and emerged flies were detected. Dead pupae were cut and the condition determined.<br />
The flies were transferred in cages and provided with tap water and a diet of yeast.<br />
Results: In these trials L. muscarium and B. bassiana were pathogenic to C. capitata after the<br />
contact of old larvae on the soil. Non dead or moulded Larvae were found in the varieties. All<br />
of the contaminated larvae could develop to pupae, but the death occurred by pupae stage. The<br />
mortality of pupae ranged between 51.6 % by L. muscarium and 46.7 % by B. bassiana in<br />
comparison to the control with 18.4 %. The development of infected flies inside the pupae was<br />
stopped 2 to 3 days after the pupation. Among of dead pupae 35 % by L. muscarium and 26.6<br />
% by B. bassiana were moulded. The fungal growth of L. muscarium and B. bassiana was<br />
clearly inside the pupae. In some cases the mycelium emerged from the pupae. This fact<br />
indicates that the spores were picked up from the soil and adhered on the body of larvae, but<br />
the infection process occurred 2 to 3 days after the pupation inside the pupae, where the body<br />
of fly was formed.<br />
Conclusions: The results showed that a soil application of L. muscarium and B. bassiana could<br />
reduce the emergency of fruit fly of C. capitata. About 48.3 % of pupae were emerged by L.<br />
muscarium and 53 % by B. bassiana, while 82 % of pupae were emerged in the control. The<br />
following experiments should be carried out to determine the effect on the eggs, pupae and the<br />
flies.<br />
Keywords: entomopathogenic fungi, Mediterranean fruit fly, mortality, Mouldiness<br />
249
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF SOIL APPLICATION OF PAECILOMYCES LILACINUS (THOM) SAMSON IN<br />
CONTROLLING MELON FLY BACTROCERA CUCURBITAE COQ.<br />
Amala, U. 1* ; Jiji, T. 1 ; Naseema, T 1,2 ; Jacob, Arthur 1 & Sheela, M.S. 1<br />
1 Department of Agricultural Entomology, College of Agriculture, Vellayani, Thiruvananthapuram 695522, India.<br />
Email: amala_uday86@yahoo.in; 2 Department of Plant Pathology, College of Agriculture, Vellayani,<br />
Thiruvananthapuram 695522, India.<br />
Background: Melon fly Bactrocera cucurbitae (Diptera: Tephritidae) is a serious pest of<br />
cucurbitaceous vegetables causing 30 to 100 per cent crop loss (Sookar et al., 2006). Annual<br />
loss in fruit and vegetable production equivalent to Rs. 26,902 million has been reported during<br />
the year 2000 in India (Stonehouse, 2001). Although management options like hydrolyzed<br />
protein spray, parapheromone trap (Zaman 1995; Stonehouse et al., 2005), field sanitation<br />
(Akhtaruzzaman et al., 2000), chemical spray (Waseem et al., 2009) have been adopted, it is<br />
ideal to develop an eco-friendly and user-friendly method for the management of fruit flies.<br />
Various Deuteromycetes fungi viz., Beauveria bassiana, Metarhizium anisopliae,<br />
Paecilomyces fumosoroseus were known to cause epizootics in fruit flies (Dimbi et al., 2003;<br />
Yee and Lacey, 2005). Jiji et al., (2006) reported the pathogenicity of B.bassiana, P. lilacinus,<br />
Aspergillus candidus: Fries on melon fly. The study was undertaken to explore the efficacy of<br />
P. lilacinus in the management of melon fly.<br />
Methods: The bio-efficacy of the entomopathogenic fungus P. lilacinus was tested on the<br />
pupae of melon flies under in vitro conditions. Glass troughs (30 cm diameter) were filled with<br />
soil to a depth of 4 cm. The soil was drenched with 30 ml of spore suspension at 1.3x10 7, 1.3<br />
x10 8 , 1.3x10 9 spores/ml. Soil drenched with sterile water served as control. Ten numbers of one<br />
day old pupae were introduced into the soils on the third, fifth and seventh day after treatment<br />
of the fungus. Observations on the number of infected pupae per trough were recorded on the<br />
tenth day. Based on in vitro studies, the fungus was tested under field conditions. Bran based<br />
formulation of the fungus was applied as soil drenching (1.3 x10 9 spores / ml) at fortnightly<br />
intervals during the fruiting period of bittergourd. Observations on the number of infected<br />
pupae / sq.m was recorded.<br />
Results: When the pupae was introduced on the third day after inoculation of the fungus<br />
significant mortality (92.45% at 1.3x10 9 followed by 72.28% at 1.3x10 8 ) was observed which<br />
was on par (69.84% at 1.3x10 7 spores/ml) with that on the tenth day after treatment. When<br />
pupae were introduced on the fifth day after inoculation of the soil with the fungus significant<br />
mortality of 69.84% at 1.3x10 7 , 74.92% at 1.3x10 8 and 92.45% at 1.3x10 9 was recorded on the<br />
tenth day. When pupae were introduced on the seventh day after inoculation of the fungus,<br />
mortality percentage of 32.30, 44.62 and 67.28 at 1.3x10 7 , 1.3x10 8 , 1.3x10 9 spores/ml,<br />
respectively was recorded on the tenth day. Under in vivo conditions the number of infected<br />
pupae varied from 2.30 to 3.99 per sq.m.<br />
Conclusions: Soil drenching with P. lilacinus is effective, eco-friendly and easily amenable<br />
method that fits well into the IPM schedule for the management of fruit flies. The same fungus<br />
was found pathogenic to B. dorsalis, the findings of which widens the scope of its practical<br />
utility.<br />
Keywords: Bactrocera cucurbitae, Paecilomyces lilacinus, melon fly, bittergourd, cucurbits
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
PERFORMANCE OF TWO INTRODUCED PARASITIODS AGAINST BACTROCERA<br />
INVADENS ON DIFFERENT HOST FRUITS AND TEMPERATURES.<br />
Appiah, Ernest F. 1, 2,* ; Mohammed, Samira A. 1 ; Ekesi, Sunday 1 & Afreh-Nuamah,<br />
Kwame 2<br />
1 ICIPE, Nairobi, Kenya. E-mail: fappiah@icipe.org; 2 Department of Crop Science, University of Ghana, Legon.<br />
Background: The horticulture industry across sub-Saharan Africa is plagued with fruit fly<br />
pests, and the situation has worsened in recent years with the arrival of the invasive Bactrocera<br />
invadens. The failure of native parasitoids to effectively control this alien species led to the<br />
introduction of two co-evolved parasitoids (the egg-prepupal parasitoid Fopius arisanus and<br />
larval-prepupal parasitoid Diachasmimorpha longicaudata) for evaluation against B. invadens.<br />
Laboratory tests showed that these parasitoids especially F. arisanus are promising candidates<br />
for management of B. invadens. However their performance on various host fruits and<br />
temperature remains to be elucidated. This information will help in guiding the releases of this<br />
parasitoid as part of IPM management strategy for B. invadens.<br />
Methods: Performance of the two parasitoids as measured by percent parasitism, total number<br />
of parasitoid progeny, sex ratio, and developmental time were assesses across four host fruits:<br />
mango, pawpaw and citrus. The suitable stage of B. invadens was exposed on the fruit peels<br />
placed on an oviposition unit to female wasps in no choice experiment. After 24 hours, the<br />
exposed hosts were incubated till parasitoid and fruit fly emergence. Total number of emerged<br />
parasitoids (male and female), their developmental time, and uneclosed puparia were recorded.<br />
Percent parasitism and sex ratio were computed for the two parasitoids on the four host fruits.<br />
The methodology was the same for the temperature experiments except that parasitized B.<br />
invadens eggs or larvae were incubated at varying temperatures.<br />
Results: Results showed that F. arisanus parasitizes eggs of B. invadens reared on mango,<br />
pawpaw and citrus resulting in a mean % parasitism of 63.8%, 62.4% and 24.0% respectively.<br />
The % parasitism for eggs reared on citrus was significantly lower compared to mango and<br />
pawpaw and this may have been due to the significantly low number of pupae recovered in<br />
citrus experiments. Parasitism rates of 34.9%, 25.4%, 23.2% and 18.4% for mango, pawpaw,<br />
guava and citrus were recorded with respect to D. longicaudata with the rate for mango being<br />
significantly higher than the other fruits. For temperature experiments, 35°C was found to be<br />
detrimental to the development of parasitized B. invadens eggs and larvae, as all eggs and<br />
larvae were unable to complete the full developmental cycle. Temperature of 25°C was the<br />
most favourable for the development and rearing of both F. arisanus and D. longicaudata.<br />
Conclusions: F. arisanus and D. longicaudata are effective biological control agents against B.<br />
invadens. With the exception of citrus which is not favourable for the development of B.<br />
invadens, the two parasitoids were effective against B. invadens eggs reared on mango and<br />
pawpaw fruits. The two parasitoids were relatively tolerant to temperature conditions that exist<br />
on the African continent.<br />
Keywords: Bactrocera invadens, Fopius arisanus, Diachasmimorpha longicaudata, host fruits, temperature<br />
251
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
RADIATION DOSES TO SAFELY RELEASE THE PARASITOID DIACHASMIMORPHA<br />
LONGICAUDATA REARED ON ANASTREPHA FRATERCULUS LARVAE.<br />
Bachmann, Guillermo; Carabajal Paladino, Leonela; Conte, Claudia; Devescovi,<br />
Francisco; Milla, Fabián H.; Cladera, Jorge L.; Segura, Diego F.* & Viscarret, Mariana<br />
M.<br />
Instituto Nacional de Tecnología Agropecuaria (INTA), De los Reseros y Repetto s/n (1712), Buenos Aires,<br />
Argentina. Email: dsegura@cnia.inta.gov.ar.<br />
Background: Anastrepha fraterculus is a fruit fly of economic importance in Argentina and<br />
other countries in South America. Biological control programs based on the release of the<br />
braconid Diachasmimorpha longicaudata have been successful to control other flies of the<br />
genus Anastrepha. In order to release this parasitoid it is necessary to establish the radiation<br />
dose that, both, avoid fruit fly emergence from non parasitized pupae and, do not affect<br />
parasitoid performance.<br />
Methods: A. fraterculus larvae were irradiated with X-ray, at doses equivalent to: 60, 80 and<br />
100 Gy (6250.2, 8333.6 and 10417.0 roentgens, respectively). Immediately after irradiation,<br />
larvae were offered to D. longicaudata mated females. A control group of irradiated larvae<br />
were not exposed to parasitoids. Number of emerged parasitoids, flies and non emerged pupae<br />
were registered. Emerged flies (if any) were crossed with non-irradiated flies and egg<br />
production and egg hatch were registered. For those parasitoids that emerged from irradiated<br />
larvae fecundity was determined. Fecundity was assessed by offering host larvae every other<br />
day, during 2 weeks.<br />
Results: Larvae irradiated at 60 Gy showed high level of emergence and were as fertile as<br />
control (non-irradiated larvae). At 80 Gy, only 1.4% of the pupae emerged, males having a<br />
significantly reduction in fertility. No flies emerged from larvae irradiated at 100 Gy.<br />
Parasitism rate on larvae irradiated at 80 or 100 Gy did not differ from that found in control<br />
(non-irradiated) larvae, nor did their offspring’s sex ratio. F1 fertility was also not affected by<br />
the radiation dose.<br />
Conclusions: These results suggest that irradiation with X-rays at a dose equivalent to 100 Gy<br />
applied to third instar larvae of A. fraterculus can significantly reduce the risk of release of<br />
adult fruit flies on the field without affecting the development and subsequent performance of<br />
D. longicaudata. This dose could be reduced to 80 Gy, as emergence will also be close to zero<br />
and emerged flies will be to a large extent sterile.<br />
Keywords: Anastrepha fraterculus, Diachasmimorpha longicaudata, Biological Control, X-ray
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
STUDY ON THE ACTIVITY OF THE PUPAL PARASITOID SPALANGIA CAMERONI<br />
(HYMENOPTERA: PTEROMALIDAE) AS A POTENTIAL BIOLOGICAL CONTROL<br />
AGENT OF THE MEDITERRANEAN FRUIT FLY CERATITIS CAPITATA (DIPTERA:<br />
TEPHRITIDAE).<br />
Beitia, Francisco 1* ; Pérez-Hinarejos, Marta 1 , Böckmann, Elias 2 ; Fischer, K 2 ; Alonso,<br />
Marina 3 ; Gayubo, Severiano F. 3 ; Asís, José D 3 . & Tormos, José 3<br />
1 Associated Unit of Entomology, Centre of Plant Protection and Biotechnology, Instituto Valenciano de<br />
Investigaciones Agrarias (IVIA), Moncada (Valencia) SPAIN. fbeitia@ivia.es 2 Ernst-Moritz-Arndt University,<br />
Departement of Zoology, Greifswald, GERMANY. 3 University of Salamanca, Faculty of Biology, Salamanca,<br />
SPAIN<br />
Background: In spite of the lasting presence of the Mediterranean fruit fly, Ceratitis capitata,<br />
in the Mediterranean area there were not references for native parasitoids of the pest in Spain.<br />
The pteromalid Spalangia cameroni, has recently been identified (in 2003) as a parasitoid of C.<br />
capitata in Spain. This insect is well known as a pupal parasitoid of flies belonging to different<br />
taxonomic families: Muscidae, Sarcophagidae and Tephritidae. But it was not known its<br />
parasitism on C. capitata until now. Due to the importance of the Medfly as a serious pest on<br />
citrus and fruit trees in the Community of Valencia (Spanish eastern coast), the parasitic<br />
activity of this parasitoid on C. capitata is being studied in laboratory conditions for the last 5<br />
years to assess its potential as a biological control agent of the Mediterranean fruit fly.<br />
Methods: In this work we present data about several parasitic features (fecundity, fertility,<br />
host-age and host number incidence, …) in the parasitism of Spalangia cameroni on pupae of<br />
Ceratitis capitata in controlled conditions (in the laboratory), as well as on the use of frozen<br />
pupae of the Medfly to be applied to the mass rearing of the parasitoid.<br />
Results: In general, results show a good activity of the parasitoid on C. capitata in the<br />
laboratory. Also, the use of frozen pupae reveals as a good method to facilitate the mass rearing<br />
of the parasitoid.<br />
Conclusions: We can conclude that this parasitoid could be an interesting candidate to be used<br />
in the biological control of the Medfly in Spain, but some work in field conditions must be<br />
done to assure its real effectiveness against the pest.<br />
Keywords: Ceratitis capitata, Spalangia cameroni, parasitic activity, mass rearing.<br />
253
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DIPLOID MALES IN DIACHASMIMORPHA LONGICAUDATA (ASHMEAD)<br />
(HYMENOPTERA: BRACONIDAE).<br />
Carabajal Paladino, Leonela 1 ; Lanzavecchia, Silvia* 1 ; Bressa, María José 2 ; Papeschi,<br />
Alba G. 2 & Cladera, Jorge L. 1<br />
1 Instituto Nacional de Tecnología Agropecuaria (INTA), Instituto de Genética “Ewald A. Favret” (IGEAF). De<br />
Los Reseros y Nicolás Repetto, Castelar, B1712WAA – Buenos Aires (Argentina). Email:<br />
lcarabajal@cnia.inta.gov.ar; 2 Universidad de Buenos Aires (UBA), Facultad de Ciencias Exactas y Naturales<br />
(FCEN), Departamento de Ecología, Genética y Evolución (EGE). Intendente Güiraldes 2160, Ciudad Autónoma<br />
de Buenos Aires, C1428EGA (Argentina).<br />
Background: Diachasmimorpha longicaudata is an endoparasitoid of last instar larva of<br />
tephritid fruit flies. It is widely used as biological control agent of these pests, and has been<br />
massively produced in several bio-factories. One of the facts that should be taken into account<br />
in order to optimize its production is the possibility of generating a bias on the sex ratio in<br />
favor of females, as they are the ones that will attack and cause the death of the host. In this<br />
direction, it is necessary to study the sex determination system in the species. Hymenopterans<br />
are characterized by being haplodiploid (females are diploid and come from fertilized eggs, and<br />
males are haploid and come from unfertilized eggs). Nevertheless, diploid males have been<br />
observed in more than 70 species. According to one of the hypotheses about sex determination<br />
in Hymenoptera, sex is determined by the status of the sex determination locus/loci. When<br />
heterozygous, the individual will develop as female, when hemizygous, as male, and when<br />
homozygous, as diploid male. Considering this, the production of diploid males should be<br />
induced by an increase in the level of endogamy. These males are considered as a genetic load<br />
for the population as they are usually infertile. They also cause enormous losses in massive<br />
production, as they come from fertilized eggs that could have become females. The ploidy<br />
level of an individual can be established by analyzing its chromosome number. According to<br />
previous cytogenetic studies performed on D. longicaudata, males have n= 20 chromosomes<br />
and females have 2n= 40 chromosomes.<br />
Methods: In order to determine whether diploid males are produced in D. longicaudata, 12<br />
crosses between virgin females and one of their haploid sons were performed. Their male<br />
descendants were studied by cytogenetics, and their ploidy level was recorded. Chromosome<br />
preparations were made by spreading of testis from prepupa, pupa and pre-emerged adults,<br />
dyed with DAPI and photographed.<br />
Results: 10 out of the 12 crosses produced male and female descendants, indicating that the<br />
female was fertilized. Per couple, between 3 and 21 individuals were karyotyped. A total of<br />
108 chromosome preparations were performed, and from them, 49 corresponded to haploid<br />
males, 6 to diploid males (2n= 40 chromosomes), 9 were recorded as haploid males but with<br />
not enough certainty, and other 9 were identified as diploid males in the same conditions. The<br />
rest of the individuals (35) didn’t show cells in metaphase stage, so it wasn’t possible to<br />
determine their ploidy level.<br />
Conclusions: These results constitute the first description of diploid males in D. longicaudata,<br />
and set a starting point to study the sex determination system in this species in order to<br />
optimize the rearing protocols applied up to now.<br />
Keywords: Biological control, sex determination, diploid male
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
INCREASE OF PARASITISM RATES OF DIACHASMIMORPHA LONGICAUDATA<br />
REARING ON X-IRRADIATED CERATITIS CAPITATA LARVAE.<br />
Conte, Claudia 1 ; Fernández Salinas, Maria L. 1 ; Lanzavecchia, Silvia B.* 1 ; Viscarret,<br />
Mariana M. 2 ; Segura, Diego F. 1 & Cladera, Jorge L. 1<br />
1 Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética “Ewald A. Favret”. De Los Reseros<br />
y N. Repetto s/n. Castelar. Buenos Aires. Argentina. Email: cconte@cnia.inta.gov.ar; 2 Instituto Nacional de<br />
Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola. De Los Reseros y N. Repetto<br />
s/n. Castelar. Buenos Aires. Argentina.<br />
Background: Control strategies including the sterile insect technique (SIT) and biological<br />
control using parasitoids are currently applied against Ceratitis capitata. In Argentina, the<br />
larval parasitoid Diachasmimorpha longicaudata is currently being evaluated as candidate for<br />
biological control. Parasitism rate is one of the most important parameter to determine<br />
productivity and efficacy of parasitoids as biocontrol agents. The study of host-parasitoid<br />
interactions provides useful information to optimize the massive rearing of parasitoids. This<br />
interaction involves the immune response which is triggered after the female parasitoid<br />
oviposits on the host larvae. Previous studies have shown that the exposure to radiation<br />
suppresses host immune system responses in other insect species. Our objective was to study<br />
parasitism rates by D. longicaudata on irradiated larvae of C. capitata.<br />
Methods: Late second stage-early third stage of C. capitata larvae were treated with different<br />
X-ray doses (20, 40 and 80 Gy) prior to exposure to D. longicaudata females. Seven replicates<br />
were tested for each dose. Parasitized larvae were kept at controlled condition of humidity and<br />
temperature until emergence. Non-irradiated larvae were used as a control. Parasitism rates<br />
were calculated from the number of emerged parasitoids for each X-ray dose. The parasitism<br />
rate was corrected taking into account the number of non-emerged parasitoids. A one-way<br />
ANOVA was performed. When the test was significant, post-hoc pair-wise comparisons were<br />
performed using Tukey`s HSD (Honestly Significant Difference).<br />
Results: Parasitism rates of larvae exposed to 20 G and 40 Gy and non-irradiated larvae<br />
showed no significant differences. Larvae irradiated with 80 Gy showed the highest rate of<br />
parasitism which significantly differed from those obtained with non-irradiated larvae (p
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
INCIDENCE OF PARASITOIDS OF ANASTREPHA SPP., IN BACKYARD OCHARDS IN<br />
THE SOUTHEAST OF CAMPECHE, MEXICO.<br />
García Ramírez, María de Jesús 1*; Medina Hernández, Rosa E. 1 , Baeza Gómez, Jenny<br />
M. 1 ; López-Martínez, Victor 2 ; Vázquez López, Margarito 1 & Encalada Mena, Lisandro<br />
A. 1<br />
1 Escuela Superior de Ciencias Agropecuarias de la Universidad Autónoma del Estado de Campeche. Calle 53 s/n<br />
Col. Esfuerzo y Trabajo N.2. C.P. 24350. Escarcega Campeche, México. 2 Facultad de Ciencias Agropecuarias.<br />
Universidad Autónoma de Morelos. mjgarcia.25@gmail.com.<br />
Background: The fruit flies of the genus Anastrepha spp. are the principal phytosanitary<br />
problem in fruit crops in tropical and subtropical areas in Latin America, this insects attack a<br />
wide range of exotic and native host plants for oviposition. The geographical position of the<br />
Mexican state of Campeche in the tropics, with its proximity to the Gulf of Mexico and the<br />
Caribbean Sea, determine the conditions for the proliferation of many species of the genus<br />
Anastrepha spp. This aspect gives us the opportunity to study commercial and native host<br />
plants for fruit flies detection in backyard orchards.<br />
Methods: Fruit collections were made in backyard orchards located in the southeastern state of<br />
Campeche, Mexico (18 º 29 '251 " N and 90 º 55' 309 W, 42.6 meters above sea level, with<br />
warm sub-humid climate and summer rains). The collection period was from June 2008 to July<br />
2009. Fruits were collected from Pouteria sapota, Manguifera indica, Prunus domestica,<br />
Citrus paradisi, Citrus aurantum, Psidium guajava, Talisia olivaeformis and Zuelenia guidonia<br />
trees. The material collected was transported to the laboratory of the Escuela Superior de<br />
Ciencias Agropecuarias, Universidad Autónoma de Campeche. The fruits were dissected and<br />
inspected for fruit flies larva presence. Third instar larvae were located in plastic container (5 x<br />
5 cm) with a mesh cover for fruit flies and parasitoids emergence.<br />
Results: All species of commercial and native fruits sampled were attacked by Anastrepha fruit<br />
flies. The species reared were A. ludens, A. serpentina, A. obliqua, A striata and A fraterculus.<br />
The larval parasitoids Doryctobracton aerolatus, Aganaspis pelleranoi and Odontosema<br />
alvinerve were reared.<br />
Conclusions: In the Mexican backyard orchards in Campeche, A. fraterculus can complete its<br />
biological cycle in many commercial and native fruit hosts. The parasitoids Doryctobracton<br />
aerolatus, Aganaspis pelleranoi and Odontosema alvinerve were associated to A. fraterculus.<br />
Keywords: Anastrepha hosts, larval parasitoids
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
TOXICITY OF SPINOSAD ON SCAPTOTRIGONA MEXICANA, MELIPONID BEES<br />
(HYMENOPTERA: APIDAE).<br />
Gómez-Escobar, Enoc¹; Guzmán-Díaz, Miguel Á.²; Flores-Breceda, Salvador¹* &<br />
Montoya, Pablo J. 1<br />
1 Subdirección de Desarrollo de Métodos, Programa Moscafrut SAGARPA-IICA. Central Poniente 14, Col.<br />
Centro.Tapachula, Chiapas 30700. Email: ²El Colegio de la Frontera Sur. Carretera Antiguo Aeropuerto Km 2.5,<br />
CP 30700, Tapachula, Chiapas, México<br />
Background: Spinosad (GF-120; Dow AgroSciences,* NF Naturalyte 0.24 g ai/L) is used to<br />
control outbreaks of Ceratitis capitata in the border Mexico-Guatemala, as part of the Medfly<br />
program activities against this pest. Although this product is considered to be selective and<br />
harmless to European bees, little is known about the impact on populations of other associated<br />
insects in fruit orchards, such as meliponid bees Scaptotrigona mexicana (Hymenoptera:<br />
Apidae), a common pollinator agent in rambutan and mango orchards in Chiapas, Mexico.<br />
Methods: This work was carried out in the gardens of Moscafrut SAGARPA-IICA Program<br />
located in Metapa de Dominguez, Chiapas, Mexico. Meliponid bee hive was provided by the<br />
laboratory of apidology of El Colegio de la Frontera Sur. Bioassays were carried out using<br />
cylindrical acrylic cage 9 cm in diameter and 8 cm high. We evaluated treatments for fresh and<br />
48 h after application of spinosad, at concentrations of 10, 20, 40, 80 ppm ai. The exposure of<br />
the product within each cage (20 mL) was carried out in a plastic slotted cylindrical piece of<br />
1.2 cm in diameter and 10 cm long. For each bioassay, ten repetitions of each concentration<br />
were performed, recording mortality at 24 and 48 h.<br />
Results: Mortality of adult meliponid exposed to spinosad residues in fresh showed no<br />
difference between the different concentrations and control. When the exposure time was<br />
increased up to 48 h no differences among treatments were obtained. Similarly, exposure of dry<br />
residues did not significantly affect the survival of insects. When the recommended dose to<br />
control fruit flies (80 ppm) was tested, mortality of meliponid bees exposed to either dry or<br />
fresh residues was not higher than in control.<br />
Conclusions: Survival of adult Scaptotrigona mexicana exposed to residues of spinosad, was<br />
not affected significantly when the product was applied according to manufacturer's<br />
recommendations.<br />
Keywords: Pollinators, stingless bees, Scaptotrigona, toxicity, Spinosad<br />
257
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
HORIZONTAL TRANSMISSION OF FUNGAL INFECTION BETWEEN CERATITIS<br />
CAPITATA ADULTS UNDER LABORATORY CONDITIONS.<br />
Imoulan, Abdessamad * & El Meziane, Abdellatif.<br />
Laboratory of Biotechnology, Valorisation and Protection of Agro-Resources, Faculty of Sciences and<br />
Technology, Cadi-Ayyad University, B.P. 549, Marrakesh, Morocco. Email: imoulan@gmail.com<br />
Background: The Mediterranean fruit fly (medfly), Ceratitis capitata, is widely distributed<br />
around the World and is regarded as the most important pest of agricultural resources. Its<br />
management is still internationally based on treatments with broad-spectrum chemical<br />
insecticides. Even though the contact pesticides are commonly effective at higher doses, they<br />
cause environmental, ecological and human health problems. Therefore, alternatives strategies<br />
have to be developed urgently to control this devastating fruit pest. Biological control by using<br />
entomopathogens could represent a suitable alternative strategy to chemical control of fruit<br />
flies. Until now, no efficient method was used for the application of entomopathogenic fungi in<br />
the field. Dissemination of conidia from infected male or female to its uninfected opposite sex<br />
among medfly population could be a potential method. Our investigation aims (i) to evaluate<br />
the possibility of horizontal transmission between medfly adults, (ii) to examine whether the<br />
medfly male or female could be an efficient spreader of conidia and (iii) to determine how<br />
horizontal transmission was affected by the infected males/females ratios and the inoculation<br />
method.<br />
Methods: Fresh conidial suspensions of Moroccan Beauveria bassiana isolates were prepared<br />
and their germination percentages were estimated. Emerged medfly adults were inoculated<br />
with 2x10 6 conidia and then coupled with clean opposite sex. Infected male insects were mated<br />
with clean females at different ratio: (i) 5 males:5 females, (ii) 5 males with 10 females, (iii) 5<br />
males with 25 females and (iv) 5 males with 50 females. Medfly Infection (mycosis) and<br />
Mortality were evaluated using wet and dry conidia.<br />
Results: Horizontal transmission of Beauveria bassiana isolates, between different sexes of C.<br />
capitata, was evaluated in the laboratory. Uninfected males and females could became infected<br />
when mated with infected opposite sex. The infection percentage of uninfected males and<br />
females ranged from 55% to 76.67% and from 58.33 to 75.00%, respectively, whereas the<br />
average of survival time (AST) varied between 11.80 to 15.10 and 13.50 to 15.34 days,<br />
respectively. Although, the males or females transmitted equally the infection to their opposite<br />
sex during mating, the infection percentage of females was found closely correlated to the<br />
ratios of inoculated males to uninfected females. Furthermore, the horizontal transmission of<br />
conidia was significantly different between B. bassiana isolates. Nevertheless, the mycosis<br />
percentage increased when dry conidia was used (66.67-100%) compared to wet conidia<br />
(26.67-93.33%). Moreover, the ASTs were shorter with dry conidia (2.36-8.62 days) compared<br />
to wet conidia method (12.37-20.06 days).<br />
Conclusion: Horizontal transmission was proved among medfly adults at the laboratory and<br />
varied significantly in relation to the B. bassiana isolates. The infection percentage and AST of<br />
uninfected females were correlated positively to the ratios of infected males. This work<br />
revealed that horizontal transmission could be a useful method to management of C. capitata in<br />
field.<br />
Keywords: Beauveria bassiana, C. capitata, horizontal transmission, biological control, Argan forests.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
CONSERVATION OF PREDACIOUS SPIDERS FOR CONTROLLING FRUIT FLIES IN<br />
MANGO ORCHARDS.<br />
Jumroenma, Kriengkrai; Vangsilabutr, Wipada; Sudhi-Aromna, Sarute; Srikachar,<br />
Sunyanee* & Plodkornburee, Wipada<br />
Entomology and Zoology Group, Plant Protection Research and Development Office, Department of Agriculture,<br />
Bangkok, Thailand. Email: nutaa2000@yahoo.com<br />
Background: Fruit flies (Diptera: Tephritidae) are the major pests of various fruits in Thailand. Two<br />
species of tephritid flies, the Oriental fruit fly, Bactrocera dorsalis and the guava fruit fly, Bactrocera<br />
correcta have been found in mango orchards all year round. The dominant species is B. dorsalis.<br />
Female fruit flies cause damage by laying their eggs in ripe or nearly ripe fruit. The larval stage lasts<br />
about one week then leaves the fruit and drops into the ground to pupate in the soil. Many species of<br />
spider role as a predacious spider in mango orchards.<br />
Methods: To study on the outbreak and population density of fruit flies in mango orchard, steiner bait<br />
traps with methyl eugenol and insecticide were placed throughout the year in mango orchard areas in<br />
Chachoengsao province. The spiders were collected by beating the brunches of mango trees and by<br />
sweeping on the weedy areas at unsprayed and spayed orchards. The species and number of fruit flies<br />
and predacious spider were surveyed and collected.<br />
Results: the Oriental fruit fly, B. dorsalis and the guava fruit fly, B. correcta were the most important<br />
fruit flies pests in mango orcahrd. Bactrocera correcta was found at the average of 9.68 - 44.73<br />
flies/trap/day during March - May 2008, the seasonal harvesting period of mango in Thailand. Sixty<br />
six species of 50 genus and 17 families of spider fauna were found inhabiting in mango orchards.<br />
The lynx spider, Oxyopes lineatipes is the most important predacious spider in consuming fruit flies.<br />
The immature stage, adult females and males of the lynx spider consume fruit flies 7.78, 7.67 and<br />
6.53 flies/day, respectively. In the orchard, spiders stay on weed waiting for newly emerged fruit flies<br />
climbing up from the ground. In the unsprayed orchard, high density of spider was found on weed<br />
under the tree canopy as well as at the water ways' side areas, whereas many spiders were found on<br />
weed at water ways' side areas in the sprayed orchard. In order to conserve the predacious spiders in<br />
mango orchards, we concluded that weeds should be kept at certain spots as to provide a shelter for<br />
them.<br />
Conclusion: Bactrocera dorsalis is the dominant fruit fly species and the lynx spider, O. lineatipes is<br />
the important natural enemy in mango orchard. The immature stage, adult females and males could<br />
consume fruit flies 7.78, 7.67 and 6.53 flies/day. To conserve the predacious spider in mango<br />
orchards, weed should be maintained at certain spots as to provide a shelter for them.<br />
Keywords: conservation, Bactrocera dorsalis (Hendel), mango<br />
259
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ANALYSIS OF TWO LARVAL-PUPAL PARASITOIDS (HYMENOPTERA,<br />
BRACONIDAE) IN THE BIOLOGICAL CONTROL OF CERATITIS CAPITATA<br />
(WIEDEMANN) IN SPANISH MEDITERRANEAN AREAS.<br />
Martins, David S. 1,2 ; Skouri, Wafa 1,3 ; Chermiti, Brahim 3 ; Aboussaid, Houda 1,4,5 ; El<br />
Messoussi, Said 4 ; Oufdou, Khalid 5 ; Carbonell, Emilio 6 , Sabater-Muñoz, Beatriz 1 &<br />
Beitia, Francisco 1*<br />
1 Associated Unit of Entomology, Centre of Plant Protection and Biotechnology, Instituto Valenciano de<br />
Investigaciones Agrarias (IVIA), Moncada (Valencia) SPAIN. fbeitia@ivia.es 2 Instituto Capixaba de Pesquisa,<br />
Assistência Técnica e Extensão Rural, INCAPER, UFV/CNPq. Vitoria, ES, BRASIL. 3 Institut Supérieure<br />
Agronomique de Chott-Mériem, Université de Sousse. 4042 Chott-Mériem TUNISIA. 4 Laboratory of Molecular<br />
Modeling and Ecophysiology, Faculty of Sciences-Semlalia, Cadi Ayyad University, Marrakesh, MOROCCO.<br />
5 Laboratory of Biology and Biotechnology of Microorganisms, Faculty of Sciences-Semlalia, Cadi Ayyad<br />
University. 6 Biometry Unit, IVIA.<br />
Background: The Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), is<br />
considered a key pest in fresh fruit and citrus production in the Mediterranean Bassin.<br />
Nowadays, it is being studied the use of several ecological methods against this pest in some<br />
Mediterranean countries (Spain, Morocco and Tunisia), like the Sterile Insect Technique (SIT),<br />
the classical biological control (CBC) and the search for native parasitoids. Two exotic larvalpupal<br />
parasitoids of fruit-flies have been imported by the IVIA to Spain: Diachasmimorpha<br />
tryoni and D. longicaudata. As it occurs in other countries, in Spain it is being studied the<br />
combined use of the SIT and overflooding releases of parasitoids to combat Medfly<br />
populations. To achieve this, only one parasitoid species is usually mass reared and released.<br />
Over the last two years we have been analyzing the competitiveness of D. longicaudata and D.<br />
tryoni in order to choose the species to be used together the SIT for the integrated control of the<br />
Medfly.<br />
Methods: Four factors have been studied in laboratory, greenhouse and field trials to select one<br />
species for mass-rearing. These factors are: i) Adult longevity at the four seasons in field; ii)<br />
Fecundity and fertility of females in the laboratory; iii) parasitism at variable host densities in<br />
laboratory; and iv) interspecific competition in laboratory and greenhouse.<br />
Results: When separately studied in the laboratory both species seems to have similar potential<br />
as parasitoids of the Medfly. But in the intraspecific competition assay D. longicaudata shows<br />
a significant more effectiveness than D. tryoni in offspring production (either in laboratory or<br />
in greenhouse experiments). This result could mean a competitive superiority of D.<br />
longicaudata. Field trials have showed an unexpected high longevity in both species but more<br />
important in D. longicaudata. The experimental releases were performed with a low number of<br />
females, but even in this conditions D. longicaudata performed better than D. tryoni, as only<br />
individuals of the previous species have been recovered.<br />
Conclusions: D. longicaudata seems to present a competitive superiority in medfly parasitism<br />
over D. tryoni in Mediterranean conditions. A joint view of all results leads us to select D.<br />
longicaudata for the application of CBC in the Comunidad Valenciana and open the possibility<br />
to use it in the other countries from the north of Africa.<br />
Keywords: Ceratitis capitata, Biological control, parasitoid efectiveness.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
INSECTICIDAL ACTIVITY OF TUNISIAN STRAINS OF BACILLUS THURENGIENSIS<br />
AGAINST LARVAE AND ADULTS OF THE MEDITERRANEAN FRUIT FLY CERATITIS<br />
CAPITATA WIEDEMAN (DIPTERA: TEPHRITIDAE).<br />
Mediouni-Ben Jemâa, Jouda 1 *; Messoudi, Soumaya 2 & Belhadj Omrane 2<br />
1 Laboratoire de Protection des Végétaux, Institut National de la Recherche Agronomique de Tunisie, Rue Hedi<br />
Karray, 2092 Ariana, Tunis, Tunisia. Email: joudamediouni@lycos.com. 2 Laboratoire de Biochimie et de<br />
Technobiologie, Faculté des Sciences de Tunis, 2092 EL Manar II, Tunisia.<br />
Background: The Mediterranean fruit fly, Ceratitis capitata is one of the key pests on citrus<br />
and several others fruit in Tunisia. Chemical control using a broad-spectrum of insecticides<br />
remains the most used tool for the management of this pest. However, problems related to the<br />
appearance of insect resistance, emergence of secondary pests, impact on non target organisms,<br />
environmental pollution and residues further restrict the use of such method. In the aim of<br />
searching effective and environment friendly alternatives, the bacterium Bacillus thuriengiensis<br />
has been successfully employed to control insect pests in the orders of Lepidoptera,<br />
coleopteran and diptera including C. capitata.<br />
Methods: The insecticidal activity of 8 Tunisian strains of Bacillus thuringiensis isolated from<br />
various regions of Tunisia and selected among 120 strains was assessed against the third instar<br />
larvae and newly emerged adults of the Mediterranean fruit fly C. capitata. The strains isolates<br />
were tested for their activities by using respectively spore-crystal in suspension and<br />
lyophilized. Two methods were used. The first method consists in pulverizing the culture<br />
medium by 4 ml of the suspension spore-crystal. The second consists in the mixture of the<br />
larvae medium by the lyophilised suspension spore-crystal with increasing concentrations:<br />
0.032; 0.048 and 0.064 mg/g of culture medium. The death rate is noted every 24 h during 3<br />
days.<br />
Results: Results showed that the suspension spores-crystal of Bt18, Bt4 and Bt21 strains were<br />
very effective against larvae with respectively death rates of 85, 90 and 95%. The strains Bt13<br />
and Bt3 present a significant death rate with respectively 65 and 70%. Newly emerged adults<br />
were less sensitive to all tested strains compared to larvae. The highest toxicity was obtained<br />
with Bt21 with 55% of death. According to the second method, highest toxicities for larvae<br />
were also obtained for Bt18, Bt4 and Bt21 strains with respectively death rate of 80, 85 and<br />
90%. For new emerged adults, highest toxicity was observed with Bt4 with 45% death rate.<br />
Conclusions: Therefore, the use of B. thuriengiensis could be integrated with some of these<br />
existing methods in an IPM approach to control C. capitata.<br />
Keywords: Bacillus thuringiensis, biopesticide, Ceratitis capitata<br />
261
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FUNCTIONAL RESPONSE AND PATCH TIME ALLOCATION OF THE FRUIT FLY<br />
PARASITOID FOPIUS ARISANUS (SONAN) ON BACTROCERA INVADENS DREW,<br />
TSURUTA & WHITE.<br />
Merkel, Katharina* 1,2 ; Ekesi, Sunday 1 & Hoffmeister, Thomas S. 2<br />
1 International Centre of Insect Physiology and Ecology (icipe), Plant Health Department, P.O. Box 30772-00100,<br />
Nairobi, Kenya. Email: kmerkel@icipe.org. 2 University of Bremen, Institute of Ecology, Population Ecology &<br />
Evolutionary Ecology, FB 2, Bremen, Germany.<br />
Background: To predict and understand the success or failure of a biocontrol agent it is<br />
necessary to study their behavior and mechanisms of decision making. In this study we<br />
investigated the functional response of the egg-prepupal parasitoid, Fopius arisanus, on<br />
Bactrocera invadens and the effect of host encounters on the patch residence time. Depending<br />
on the distribution of hosts in the field, successful host encounters can have either an<br />
incremental or decremental effect on patch residence time. Fopius arisanus is known to<br />
discriminate between suitable and unsuitable hosts. Hence, encounters with parasitized or<br />
unhealthy hosts should lead to a decrease in the patch residence time.<br />
Methods: Seven-day old, experienced females of F. arisanus were individually exposed to<br />
artificially infested mango domes (3, 6, 9, 18, 27, 54 or 81 eggs of B. invadens). The behavior<br />
of the parasitoids was recorded until the time the female left the mango dome using The<br />
Observer® 2.0 software. To estimate the parasitization rate, host eggs were dissected and the<br />
number of parasitized eggs recorded. Since the host density was gradually depleted over time<br />
the data were fitted to Royama’s equation. The effect of oviposition on the patch leaving<br />
tendency was tested using the Cox proportional hazards model.<br />
Results: The efficiency of F. arisanus increased from 5.5 to 14.9 parasitized hosts per hour at a<br />
host density of 3 and 81, respectively. The mean residence time increased from 1782s for the<br />
lowest to 7069s for the highest host density. Results also showed that oviposition events<br />
decreased the patch leaving tendency (exp(β) = 0.67, P < 0.0001) hence increased the<br />
responsiveness of the parasitoid towards the patch. According to Royama’s model the handling<br />
time was 198.8s while from our observations one oviposition took an average of 80.5s.<br />
Conclusions: Our results suggest that F. arisanus should be able to contribute to the<br />
management of B. invadens. The practical application of this finding with regard to field<br />
suppression of the pest will be presented.<br />
Keywords: Bactrocera invadens, Fopius arisanus, functional response, patch time allocation
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EFFECT OF SOIL PHYSICAL FACTORS ON THE FATE AND VIRULENCE OF THE<br />
MITOSPORIC ASCOMYCETE METARHIZIUM ANISOPLIAE AGAINST PUPARIA OF<br />
CERATITIS CAPITATA.<br />
Quesada-Moraga, Enrique*; Torrent, J.; Barrón, V.; Santiago-Álvarez, Cándido &<br />
Garrido-Jurado, I.<br />
Agricultural Entomology Unit. Department of Agricultural and Forestry Sciences, University of Cordoba. Campus<br />
de Rabanales C4, 2º planta 14071 Córdoba, Spain. Email: cr2qumoe@uco.es.<br />
Background: As an alternative to chemical control or as part of IPM programs, there is a<br />
resurgence of interest in the use of microbial insecticides for biological control of tephritid<br />
flies, particularly entomopathogenic fungi (EF). The strategic options in using<br />
entomopathogenic fungi in medfly control are aerial applications and autodissemination<br />
targeting adults and soil inoculation targeting pupariating larvae and puparia. With this aim, in<br />
a previous study, we have found Metarhizium anisopliae Sorok EAMa 01/58-Su isolate to be<br />
highly virulent against both C. capitata adults and puparia. However, factual information on<br />
the fate of the conidia after they reach the soil will provide a better understanding of the<br />
potential of this M. anisopliae strain for being applied as an effective biological insecticide<br />
against medfly puparia. In this work, we report on the effect of soil pH and texture on the fate<br />
in the soil and virulence of EAMa 01/58-Su strain against medfly puparia.<br />
Methods: Sixteen representative Mediterranean soils were used for the studies the effect of soil<br />
type on the adsorption of conidia on the soil. The time course evolution of the number of<br />
available conidia in each soil suspension was obtained either under standard saline conditions<br />
or flocculation conditions. Then, the movement of conidia in the soil was studied using soil<br />
columns with four soils representing sandy acid and alkaline and clay acid and alkaline<br />
conditions respectively. These four soils were also used for evaluating the physical conditions<br />
of the soil on the virulence of EAMa 01/58-Su strains against medfly puparia in transparent<br />
containers (8 by 8 by 5.5 cm), each holding 30 g of soil.<br />
Results: Our results indicate that increasing clay contents in the soil resulted in lower number<br />
of available conidia in the soil solution. Even if in the soil columns most of the conidia were<br />
retained in the first soil layer for all soils, this was particularly significant in the sandy ones.<br />
Interestingly, the soil salinity clearly influenced the adsorption on the conidia in the soil.<br />
Besides, virulence of this strain against medfly puparia in the soil was also depended on the<br />
type of soil.<br />
Conclusions: Our results show the potential of soil treatments with M. anisopliae EAMa 01/58-<br />
Su strain for the microbial control of medfly puparia even if the soil physical conditions have<br />
to be considered to decide on the number of application etc. This strain could be used therefore<br />
in a biocontrol strategy targeting both adults in aerial sprays and to the soil beneath the tree for<br />
controlling puparia, the soil-inhabiting stage.<br />
Keywords: Medfly, Metarhizium, Beauveria, soil application, geodesinfestation, puparia<br />
263
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE STERILE INSECT TECHNIQUE AND PARASITOIDS (HYMENOPTERA:<br />
BRACONIDAE) FOR THE CONTROL OF THE QUEENSLAND FRUIT FLY,<br />
BACTROCERA TRYONI (FROGGATT) (DIPTERA: TEPHRITIDAE).<br />
Reynolds, Olivia L. 1 *; Jessup, Andrew J. 2 ; & Gurr, Geoff M. 3<br />
1 EH Graham Centre for Agricultural Innovation, Industry & Investment New South Wales, Private Bag 4008,<br />
Narellan NSW, Australia 2567, Email: olivia.reynolds@industry.nsw.gov.au; 2 Insect Pest Control Laboratory,<br />
Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency,<br />
Vienna, Austria; 3 EH Graham Centre for Agricultural Innovation, Charles Sturt University, Orange, NSW,<br />
Australia 2800.<br />
Background: The Queensland fruit fly, Bactrocera tryoni is the most significant pest of edible<br />
fruit and fruiting vegetables in Australia. With a decreasing reliance on insecticides and an<br />
increase in organic produce, biologically safe options are the way forward.<br />
Methods: The sterile insect technique (SIT) is an environmentally friendly option to control or<br />
suppress fruit fly populations or outbreaks and has been used in pest free areas (PFAs) and<br />
surrounding buffer zones to target outbreaks and suppress populations respectively. However,<br />
international studies have shown that the augmentive release of opiine braconids can suppress<br />
fruit fly populations to a greater extent than either technique alone. Parasitoids and sterile<br />
insects have the advantage to go into areas where other techniques cannot be applied.<br />
Results: This paper reports on the progress of SIT and parasitoids for the control of B. tryoni in<br />
PFAs and bordering zones. Methods for rearing and releasing sterile B. tryoni have been<br />
improved and include studies to improve the performance of the fly. Alternative release<br />
methods are also being developed and include chilled adult release. Two species of parasitoids,<br />
Diachasmimorpha kraussii and D. tryoni, not previously recorded in inland New South Wales<br />
have been identified, although in numbers insufficient to suppress B. tryoni populations to<br />
below economically acceptable thresholds. However, these results suggest that these two<br />
species are candidates for augmentative release in the region. Trials looking at the biology and<br />
ecology of these parasitoids in relation to their potential as biological control agents for B.<br />
tryoni are ongoing.<br />
Conclusions: Overall, results will be presented in relation to the potential for incorporation of<br />
the combined use of SIT and parasitoids into pest fruit fly management programs.<br />
Keywords: opiine, natural enemy, pest free area
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
SPECIES-SPECIFICITY OF A PCR-BASED ASSAY FOR IDENTIFICATION OF<br />
CERATITIS CAPITATA (WIEDEMANN) PARASITOIDS.<br />
San Andrés, Victoria 1,2* ; Castañera, Pedro 1 & Sabater-Muñoz, Beatriz 2 .<br />
Unidad Asociada de Entomología IVIA-UJI-CIB CSIC. 1 Centro de Investigaciones Biológicas (CIB) del Consejo<br />
Superior de Investigaciones Científicas (CSIC), Departamento Biología de Plantas, Ramiro de Maeztu 9; 28040-<br />
Madrid, Spain. Email: sanandres_vic@gva.es. 2 Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. de<br />
Moncada a Náquera km. 4,5; Moncada 46113- Valencia (Spain).<br />
Background: The Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) is a key<br />
pest of citrus and many other fruits. Two exotic braconid medfly parasitoids,<br />
Diachasmimorpha tryoni (Cameron) and D. longicaudata (Ashmead) were imported into<br />
Spain, and are being considered for its implementation jointly with the ongoing medfly sterile<br />
insect technique program in Valencia. Currently the identification of parasitoids and estimation<br />
of parasitism rates rely on tedious and time-consuming dissection and rearing methods. As a<br />
result, there has been increasing interest in the use of molecular methods to identify parasitoids<br />
and to assess parasitism rates. The present study describes species-specific PCR primers, based<br />
on the ribosomal internal transcribed spacer 1 region sequence (ITS-1), for three species of<br />
medfly parasitoids and their use in a multiplex PCR assay.<br />
Methods: Complete ITS sequence has been obtained for several species of parasitoids by<br />
amplification with ITS universal primers. Species-specific primers for these species and for C.<br />
capitata were designed on ITS-1 to use in combination with a reverse universal primer.<br />
Multiplex PCR conditions have been set up with parasitoid DNA mixtures. Laboratory<br />
experiments have been set up to test the multiplex PCR protocol for each species individually<br />
and in competition. Parasitism rates were estimated by both molecular and rearing and<br />
dissection methods under the conditions tested.<br />
Results: The primers set developed amplify exclusively sized species-specific PCR products in<br />
a single reaction. Sensibility of D. tryoni and D. longicaudata primers have been established at<br />
1 ng of parasitoid DNA (mixed DNA samples). In laboratory experiments, D. tryoni and D.<br />
longicaudata DNA were detected in C. capitata pupae after 48h of parasitism.<br />
Conclusions: The multiplex system developed allows establishing the parasitoid species and<br />
the parasitism rate, in a feasible fast manner.<br />
Keywords: Medfly, parasitoids, ITS-1<br />
265
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ASSESMENT OF THE POTENTIAL NON TARGET EFFECTS OF METARHIZIUM<br />
ANISOPLIAE (SOROKIN) ON THE BRACONID PARASITOIDS DIACHASMIMORPHA<br />
TRYONI (CAMERON) AND D. LONGICAUDATA (ASHMEAD).<br />
San Andrés, Victoria 1,2* ; Moya, Pilar 3 ; Ayala, Ildefonso 3 , Abad, María 3 ; Primo, Jaime 3 &<br />
Castañera, Pedro 2<br />
Unidad Asociada de Entomología IVIA–UJI-CIB CSIC 1, 2 . 1 Instituto Valenciano de Investigaciones Agrarias<br />
(IVIA), Ctra. de Moncada a Náquera km. 4,5; Moncada 46113- Valencia, Spain. 2 Centro de Investigaciones<br />
Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC), Departamento Biología de Plantas,<br />
Ramiro de Maeztu 9; 28040- Madrid, Spain. Email: sanandres_vic@gva.es; 3 Centro de Ecología Química<br />
Agrícola (CEQA), Universidad Politécnica de Valencia (<strong>UPV</strong>), Camino de Vera s/n. 46022. Valencia, Spain.<br />
Background: The demand of insecticide free fresh fruit is encouraging the use of ecologically<br />
friendly alternatives against the Mediterranean fruit fly (medfly) Ceratitis capitata<br />
(Wiedemann), such as the sterile insect technique (SIT) and biological control programs.<br />
Consequently, two imported braconid species Diachasmimorpha tryoni (Cameron) and D.<br />
longicaudata (Ashmead) started to be released in Valencia in autumn 2008. In addition, the<br />
utilization of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) Sorokin is<br />
being considered jointly with the ongoing SIT program to control medfly in the Comunidad<br />
Valenciana. Accordingly, the potential non-target effects of this pathogen on the released<br />
parasitoids need to be evaluated.<br />
Methods: Laboratory assays were performed to assess the pathogenicity of M. anisopliae to<br />
both parasitoids by exposing them to mycosed cadaver flies. The longevity of both exposed<br />
and non-exposed parasitoids was determined. Moreover, the effect of M. anisopliae on<br />
fecundity, sex ratio and emergence rate of parasitoids was calculated. Multiplex-PCR on the<br />
non-emerged pupae of C. capitata was used to assess the actual rate of parasitism.<br />
Results: The exposition of the two parasitoids to mycosed flies resulted in D. tryoni and D.<br />
longicaudata differential pathogenicity. For D. tryoni, the parasitism rate was not affected<br />
regardless of the amount of conidia received. For D. longicaudata, fecundity, and parasitism<br />
rates were affected when higher conidia contact occurred. In any case, the sex ratio of both<br />
parasitoids was affected by M. anisopliae exposure.<br />
Conclusions: This laboratory evaluation reveals that both parasitoid species are susceptible to<br />
M. anisopliae, though further field-cage and open field studies are required to confirm these<br />
results.<br />
Keywords: Medfly, parasitoids, entomopathogenic fungus, SIT
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
MEDFLY IMPACT IN SOUTHWESTERN MOROCCO AND BIOLOGICAL CONTROL<br />
WITH HALOPHYTES SPECIES.<br />
Soummane, Hassna 1,3 ; Naamani, Khalid 1* ; Larhsini, Mustafa 1 ; El Hadrami, Ismaïl 1 ;<br />
Guerfali, Meriem M'saad 2 & Coll Toledano, Josep 3 .<br />
1 Cadi Ayyad University, Faculty of Sciences Semlalia, Department of Biology, Laboratory of Biotechnologies,<br />
Protection and Valorization of Vegetables Resources, PB: 2390. Marrakesh 40000 Morocco. E-mail:<br />
knaamani@yahoo.fr. 2 National Centre of Sciences and Nuclear Technologies (CNSTN), Technopole of Sidi<br />
Thabet, 2020 Sidi Thabet, Tunisia. 3 Dep. Química Biològica i Modelització Molecular, IQAC (Institut de Química<br />
Avançada de Catalunya), CSIC. Jordi Girona 18-26 08034 Barcelona. Spain.<br />
Background: The medfly attack reduces growth capacity of the crops and therefore lower<br />
commercial yield. This effect is most important when the attacked species is considered such<br />
as high importance economic species, such as Argan in southwestern Morocco. The Argan tree<br />
(Argania spinosa) is endemic to semi-desert Souss valley of southwestern Morocco. In this<br />
area, it plays major ecologic and socio-economic roles. The argan oil is considered as a highvalue<br />
product for that it’s crucial to women’s co-operatives in Southwest of Morocco. This<br />
economic contribution depends of argan tree productivity which faces to abiotic and biotic<br />
factors, especially the dipterans fly: Ceratits capitata. The argan tree is considered as the<br />
largest fruit fly host knows in the world.<br />
Methods: In order to control this insect we propose a biological one with three halophytes<br />
species belongings to two different families. The plant extracts were sequentially partitioned<br />
for subsequent bioassay. The active fractions were divided into subfractions using a silica gel<br />
column (reverse phase). Toxicity and effectiveness of the subfractions were reported as LC50<br />
and LC90, which represent the concentrations in µg/ml with 50% and 90% medly mortality in<br />
24h, respectively. Both larvae and adult medfly mortality were considered.<br />
Results: We noted that a great insecticidal activity for Tamaricacea species comparing with the<br />
studied Chenopodiacea species. The Tamarix species showed a strongest larvicidal activity<br />
against the second and third instar larvae of Ceratitis capitata, two times greater comparing<br />
with the other extracts. These same findings were also registred for the adulticidal activity of<br />
Tamarix species it was 21.3% greater comparing with the tested Chepodiacea species.<br />
Conclusion: These results suggest that tested Tamarix species may be considered as an<br />
interesting bioinsecticidal product. Application of these products could be very useful to reduce<br />
the larvae and adult of Ceratitis capitata. Future investigations should aim at testing these<br />
products against other fruit fly species and try to resolve the mode of action of these<br />
bioinsecticidal compounds.<br />
Keywords: biological control, halophytes, Ceratitis capitata, Argania spinosa<br />
267
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
LONGEVITY OF THREE SPECIES OF TEPHRITID (DIPTERA) FRUIT FLY<br />
PARASITOIDS (HYMENOPTERA: BRACONIDAE: OPIINAE) PRESENTED WITH TWO<br />
FRUIT-BASED DIETS.<br />
Stuhl, Charles 1,2 ; Cicero Jarado, Lizzette 3 ; Sivinski, John 2* ; Aluja, Martin 3 & Teal,<br />
Peter 2<br />
1 Department of Entomology and Nematology, University of Florida, Gainesville, FL. 2 USDA-ARS, Center for<br />
Medical, Agricultural and Veterinary Entomology, Gainesville, FL. Email: John.Sivinski@ARS.USDA.GOV;<br />
3 Instituto de Ecologia, A.C., Apartado Postal 63, 91000 Xalapa, Veracruz, Mexico.<br />
Background: Opiinae braconids are important natural enemies of Anastrepha spp. and massreared<br />
in several locations for area-wide control. Adult parasitic Hymenoptera often feed on<br />
floral and extrafloral nectars and honeydews, but an Old World opiine introduced to the<br />
Americas, Diachasmimorpha longicaudtata, survives on orange juice (Citrus aurantium) as it<br />
seeps from injured fruit. The New World species, Doryctobracon areolatus and Utetes<br />
anastrephae, are also prominent Anastrepha parasitoids. The orientation to and longevity upon<br />
diets based on two different fruits were compared and contrasted in all three parasitoids.<br />
Methods: Pulp and juice were provided from a domesticated Old World fruit (C. aurantium)<br />
that offered little opportunity for shared evolutionary histories, except perhaps with D.<br />
longicaudata. All three parasitoids were also provided with a less-domesticated New World<br />
fruit (guava, Psidium guajava) which shared an evolutionary history with D. areolatus and U.<br />
anastrephae.<br />
Results: Both sexes of D. longicaudata, from USA and Mexican-derived cultures died when<br />
provided P. guajava pulp or juice at a rate similar to a water-only control. In the USA, D.<br />
areolatus and U. anastrephae, presumably adapted to the nutrient/chemical constituents of P.<br />
guajava, also died at a rate similar to that of the water-control. Survival of all three species on<br />
C. aurantium pulp and juice was greater than on water and longevity often equaled that<br />
obtained on a honey+water solution provided as a positive control. The differences in survival<br />
on the two fruits suggested that P. guajava was either: 1) innutritious, 2) toxic or 3)<br />
unattractive. Fructose found in P. guajava sustained both male and female D. longicaudtata.<br />
Starved female D. longicaudata and D. areolatus could be found at P. guajava feedingstations,<br />
but when given a choice between fruit both sexes of all species were generally found<br />
at C. aurantium stations. Thus P. guajava contained nutrient sugar and could be located over a<br />
distance of at least 15 cm by females of two species. Longevity differences between insects fed<br />
on P. guajava and C. aurantium were most likely due to an undiscovered toxin.<br />
Conclusions: Given the differences in fruit-food quality, adult opiine food sources would not<br />
be obtainable at all oviposition sites and more additional foraging for food than previously<br />
postulated may be required. Volatiles present in C. aurantium, but not P. guajava, may be<br />
candidate attractants.<br />
Keywords: Diachasmimorpha, Doryctobracon, Utetes, Psidium, Citrus
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DISPERSAL OF STERILE MALES OF ANASTREPHA LUDENS (DIPTERA:<br />
TEPHRITIDAE) TREATED WITH BEAUVERIA BASSIANA.<br />
Toledo, Jorge 1, *; Novelo-Rincón, Luisa F. 1 ; Montoya, Pablo 2 ; Oropeza, Azucena 1 ; Flores-<br />
Breceda, Salvador 2 & Liedo, Pablo 1<br />
1 Departamento de Entomología Tropical. El Colegio de la Frontera Sur (ECOSUR) Apartado Postal 36.<br />
Tapachula, Chiapas, 30700 México; Email: jtoledo@ecosur.mx; 2 Subdirección de Desarrollo de Métodos.<br />
Programa Moscamed, SAGARPA. Central Poniente # 14. Tapachula, Chiapas, 30700 México.<br />
Background: Laboratory and field cage studies have shown that sterile males can be used for<br />
the horizontal transmission of entomopathogenic fungi, as an additional mortality factor for the<br />
control of fruit fly pests. Our goal in this study was to investigate the survival and dispersal of<br />
sterile Anastrepha ludens males treated with Beauveria bassiana conidia under natural field<br />
conditions.<br />
Methods: Two central point release-recapture experiments were carried out. In the first, marked<br />
sterile A. ludens adults treated with three different concentrations of dry B. bassiana conidia<br />
and untreated sterile flies were released in a central point in a mango orchard and subsequently<br />
recaptured in a trapping net around the release point using Multilure traps. In the second<br />
experiment, only one conidia concentration was used to treat sterile flies. Field survival and<br />
dispersal was estimated from the number of trapped flies.<br />
Results: In the first experiment, the recapture of males treated with low and intermediate<br />
concentrations was similar to the recapture of untreated males. The highest conidia<br />
concentration resulted in the lowest recapture. However, the dispersal pattern was similar for<br />
all treatments. In the second experiment, the recapture of sterile untreated males was<br />
significantly greater than the recapture of treated males. The longest distance displaced was<br />
203 and 218 m for treated and untreated males, respectively; but the difference were not<br />
significant. Dispersal pattern of both, treates and untreated males was to the south-southeast<br />
area. The survival to both, treated and untreated males was similar. Only in one, out of five<br />
replicates, the difference was significant.<br />
Conclusions: Horizontal transmition of entomopathogenic fungi by sterile males is feasible.<br />
Using low conidia concentrations, the field survival and dispersal of treated males was similar<br />
to that of untreated males. These and previous results on mating competitiveness suggest that<br />
the performance of conidia treated sterile males is not affected. This new approach could result<br />
in a more effective use of the Sterile Insect Technique for the control of fruit fly pests, over all<br />
when only males are reared in the genetic sexed strain. Methods need to be developed for<br />
conidia application just before release of sterile flies.<br />
Keywords: Beauveria bassiana, horizontal transmission, microbial control.<br />
269
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GF-120 ® (SPINTOR-CEBO ® ), AN INTERESTING ALTERNATIVE IN SUSTAINABLE<br />
AGRICULTURE FOR THE CONTROL OF CERATITIS CAPITATA & BACTROCERA<br />
OLEAE. SIDE-EFFECTS ON BENEFICIALS.<br />
Viñuela, Elisa 1* & Torné, María 2<br />
1 Protección de Cultivos. E.T.S.I. Agrónomos. 28040-Madrid. (Spain). Email: elisa.vinuela@upm.es. 2 Dow<br />
Agrosciences Ibérica, Madrid (Spain).<br />
Background: Spinosad is a microbial pesticide isolated by fermentation from a soil<br />
actinomycete, with a new mode of action on the nervous system (prevention of closing of the<br />
nicotinic acetylcholine receptors), high efficacy against tephritid flies and good environmental<br />
performance, so it has been authorized in EU Organic Agriculture in 2008. Spintor-Cebo ® ,<br />
incorporates the active ingredient in low concentration (0.024%) plus a bait (mixture of several<br />
components) and it has got definitive registration from the Spanish Ministry of Environment,<br />
Rural and Marine affaires since 2009, being currently used for the control of two economically<br />
important fruit flies, the Med fly in citrus orchards and the olive fruit fly. The insecticide is<br />
applied as ultra low volume bait spraying (water amount under 10 l/ha) to the top of the canopy<br />
of trees aiming at controlling adults, and the percentage of the treated surface in aerial<br />
applications varies from 25-40% depending on the crop, while in ground applications may<br />
cover among 5-10% of the vegetation<br />
Methods: Based on the available information in literature, a summary of its characteristics,<br />
performance on fruit flies as well as its side-effects on different developmental stages of the<br />
beneficial organisms compared with that of classical pesticides is presented here. Results are<br />
mainly based in its residual contact activity on laboratory, semi-field or field and on its<br />
persistence, but some information on its toxicity by other exposures routes is presented as well.<br />
Results: Spinosad is a bit more slow acting than former active ingredients used for the control<br />
of these two fruit flies (organophosphates and pyrethroids), but its efficacy after 72 h<br />
application is comparable and the pre-harvest intervals are very short: 1 day in citrus orchards<br />
and 7 days in olive groves, because it degrades quickly. The insecticide is compatible with<br />
many enemies, mainly predators belonging to different orders and families and pollinators<br />
(honeybee), and it possess a risk to many parasitoids, but age residues are usually not harmful<br />
because photolysis is the main way of degradation.<br />
Conclusions: Based on its good environmental performance and efficacy against tephritids, as<br />
well as its compatibility with non target arthropods and pollinators, it is a good candidate to be<br />
used in the control of Ceratitis capitata in citrus orchards and for the control of the Bactrocera<br />
oleae in olives, in substitution of conventional products especially in Organic Agriculture<br />
where only natural insecticides are permitted.<br />
Keywords: Medfly, olive fruit fly, spinosad bait, side-effects, parasitoids, predators
Session 8<br />
Chemical Ecology, Attractants &<br />
Other Control Methods
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EFFECTIVENESS OF THE MASS TRAPPING TECHNIQUE IN COMBINATION WITH A<br />
NEW INSECTICIDE IN CERATITIS CAPITATA MANAGEMENT.<br />
Acín, Patricia* 1 ; Roura, Lídia 1 ; Escudero-Colomar, Adriana 2 ; Du Fretay, Gäel 3 &<br />
Palència, Jaume 1 .<br />
1 Sociedad Española de Desarrollos Químicos S.L., Marie Curie, 33; 08210 Barberà del Vallès; Barcelona<br />
(Spain). Email: pacin@sedq.es. 2 IRTA-Estació Experimental Agrícola Mas Badia, Girona (Spain). 3 ABER<br />
Consulting, Martillac (France).<br />
Background: Mass trapping has been widely used in many Mediterranean areas to control the<br />
Mediterranean fruit fly (Medfly) Ceratitis capitata giving satisfactory results when used in<br />
combination with the insecticide dichlorvos (DDVP). Due to the new directive 91/414/CEE the<br />
utilization of DDVP inside the traps has been banned except in exceptional authorizations.<br />
Therefore a new effective insecticide is needed in order to assure the success of mass trapping<br />
in this fly species. Here we report the efficacy of another insecticide, deltamethrin, in the<br />
Medfly mass trapping.<br />
Methods: Four trials have been performed in different fruit varieties and localities. The assays<br />
consisted in the deployment of a certain number of traps baited with a suitable attractant and<br />
the insecticide replacing the DDVP applied in the upper part of the trap. All traps were placed<br />
in external branches in the southern face of the trees following a homogeneous distribution in<br />
the correspondent test plot. Every week a fixed number of traps were checked where the total<br />
number of adults captured inside were evaluated differentiating between sexes in most cases. In<br />
addition, a fruit damage assessment was performed.<br />
Results: The trials carried out supposed both low and high Medfly population densities. In<br />
general, females were captured in a higher proportion than males (65-75%) except at the end of<br />
the assays where the percentage of males slightly increased. Furthermore, in those tests where<br />
the number of live flies captured in the traps was evaluated a very low proportion of them were<br />
observed (approximately 2%), showing this the effectiveness of the deltamethrin. The fruit<br />
damage rate in all experiences was rather acceptable, especially in those plots where a high<br />
number of traps were positioned.<br />
Conclusions: The efficacy of mass trapping using deltamethrin as a DDVP substitute has been<br />
proved in all tests executed. This system has a notable capability for capturing adults in both<br />
low and high Medfly populations emphasizing an appropriate female percentage caught in the<br />
traps, a general low number of live adults in the devices and a very satisfactory fruit damage<br />
percentage. Therefore this mass trapping system comprises a good alternative to the traditional<br />
DDVP use and thoroughly compatible with integrated pest management.<br />
Keywords: Medfly, mass trapping, DDVP, deltamethrin<br />
273
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
VOLATILE EMISSIONS OF CERATITIS CAPITATA (WIEDEMANN): INFLUENCE OF<br />
FLY SEX, AGE AND MATING STATUS.<br />
Alfaro Cañamás, Cristina*; Vacas, Sandra; Navarro-Llopis, Vicente; Primo, Jaime.<br />
Centro de Ecología Química Agrícola - Instituto Agroforestal del Mediterráneo. Universidad Politécnica de<br />
Valencia. Camino de Vera s/n. 46022. Valencia. Spain. calfaro@ceqa.upv.es<br />
Background: The courtship behavior and pheromone communication between males and<br />
females Ceratitis capitata have been studied for many researches, however, the sex pheromone<br />
composition is still a controversial subject. The use of semiochemicals is being extended in the<br />
control of Mediterranean fruit fly, so the research of new products affecting the medfly<br />
behavior or the composition of the sex pheromone would be interesting to improve these<br />
control methods.<br />
Methods: The experimental design to study volatile compounds emitted by C.capitata<br />
consisted of three factors at two levels: sex (male or female), mating status (virgin or mated)<br />
and age (3 or 9 days old). Volatile was collected by solid phase microextraction and the<br />
compounds were isolated and identified by gas chromatography coupled with mass<br />
spectrometry. The data were analyzed by Principal Component Analysis in order to obtain<br />
clusters of compounds with a similar emission pattern. A 3-way ANOVA was applied in order<br />
to study in detail the emission pattern of these representative compounds according to sex, age<br />
and mating status.<br />
Results: Seventy compounds emitted by C. capitata were detected. All compounds were<br />
detected in males whereas only 37 were found in female volatile profile. The most abundant<br />
compounds detected in the male volatile profile were 2-hexenoic acid, geranyl acetate, ethyl<br />
(3E)-3-octenoate and (E,E)-α-farnesene. In female volatile profile, 2-ethylhexanoic acid,<br />
trimethylamine, 2,5-dimethylpyrazine and 2-hexenoic acid were the most abundant released<br />
compounds.<br />
Conclusions: The compounds were grouped in 16 clusters, determining the emission pattern for<br />
everyone. The knowledge of the emission pattern for each compound and which of them have<br />
similar profile would be interesting to design the biological assays and the combination of<br />
compounds to determine the effect on the Mediterranean fruit fly behavior.<br />
Keywords: Ceratitis capitata, volatile profile, solid phase microextraction, pheromone.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FACTORS WHICH INFLUENCE THE EFFICACY OF MASS-TRAPPING TO CONTROL<br />
THE MEDFLY CERATITIS CAPITATA (WIEDEMANN).<br />
Alonso-Muñoz, Andrés & Garcia-Marí, Ferran<br />
Instituto Agroforestal Mediterráneo, Universidad Politécnica de Valencia, Camí de Vera 14, 46022 Valencia,<br />
(Spain). Email: fgarciam@eaf.upv.es.<br />
Background: We analyzed the spatial distribution of adult Ceratitis capitata (Diptera:<br />
Tephritidae) captured in the traps, in Citrus orchards where mass trapping was applied to<br />
control the pest, to understand how the method works, to determine the factors which influence<br />
it and to suggest changes which could improve its efficacy.<br />
Methods: In Ibiza Island, from 2006 to 2008, we established a mass trapping system of control<br />
in 31 citrus orchards. The traps, of the tephri-trap type and baited with Tripack as attractant,<br />
were uniformly distributed at a density of 50 per hectare following the standard<br />
recommendations. In the orchards, all traps of the mass trapping system were sampled at<br />
intervals of 30 to 45 days, by counting all flies captured on each trap. In all, 110 samplings,<br />
with 102 traps observed per sampling on average, were carried out.<br />
Results: The establishment of the network of traps causes usually a barrier effect, with<br />
reduction in medfly captures in a gradual and rather constant rate from the periphery to the<br />
interior of the orchards. The amount of reduction observed is usually low, since the average<br />
captures were 3.5 flies per trap and day (ftd) on the external perimetral layer of traps and 2.3<br />
ftd on the second layer of traps, reaching 0.7 ftd on the sixth layer located 70 meters inside the<br />
orchard. The following factors were analyzed for the barrier effect on medfly populations: size<br />
and shape of the orchard, presence of mature fruits, species and/or variety of fruit, month of the<br />
year, time elapsed since the mass trapping was installed, and overall medfly abundance in the<br />
periphery. Size and shape of the orchard, and month of the year, showed significant influence:<br />
Larger plots and plots with minimum ratio perimeter/area showed higher capacity in reduction<br />
of captures, and the efficiency of the system of control increases in June and July.<br />
Conclusions: According to these results, the success of mass trapping can be measured by the<br />
capacity to get high levels of reduction between the external and the internal layer of traps in<br />
the plot. The mass trapping technique must be applied in large areas, of several tens of<br />
hectares, in surfaces with a compact shape in order to minimize the perimeter, and increasing<br />
the density of traps in the periphery of the protected area. The traps should be installed when<br />
fruit ripening approaches but not before, as no improvement was observed when the period of<br />
establishment of the mass trapping in the orchards increases.<br />
Keywords: Ceratitis capitata, spatial distribution, mass-trapping, Citrus, traps<br />
275
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DIFFERENCES IN LAMBDA-CYHALOTHRIN SUSCEPTIBILITY AMONG FIELD AND<br />
LABORATORY POPULATIONS OF CERATITIS CAPITATA (WIEDEMANN).<br />
Arouri, Rabeh* 1 ; Perera, Nathalie 1 ; Beroiz, Beatriz 1 ; Hernández-Crespo, Pedro 1 ; Ortego,<br />
Félix 1 & Castañera, Pedro 1<br />
1 Centro de Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones Científicas (CSIC),<br />
Departamento de Biología Medioambiental, Madrid, Spain.<br />
Background: The Mediterranean fruit fly (Medfly), Ceratitis capitata (Wiedemann), is one of<br />
the most economically important pests of citrus orchards in Spain. The intensity of insecticide<br />
treatments with malathion-lure against C. capitata has resulted in the development of resistant<br />
populations in some areas of Spain. The situation changed in 2009, since the decision of noninclusion<br />
of malathion in Annex 1 of the Directive 91/414/EEC has resulted in the withdrawal<br />
of authorisations for plant protection products containing this insecticide. Several insecticides<br />
are approved for Medfly control in citrus crops, but spinosad and lambda-cyhalothrin are the<br />
most widely applied alternatives. It is therefore necessary to evaluate the susceptibility to<br />
lambda-cyhalothrin of field and laboratory populations in order to determine the effectiveness<br />
of this insecticide for Medfly control.<br />
Methods: C. capitata infested fruits were collected in commercial fields from different<br />
geographical areas in Spain during 2009. The susceptibility to lambda-cyhalothrin (10% w/v,<br />
Karate Zeon, Syngenta Agro S.A., Madrid, Spain) of emerging adults from each field was<br />
determined by dose-response feeding bioassays and compared to the susceptibility of a<br />
malathion-resistant and a susceptible laboratory strain. In addition, the malathion-resistant<br />
strain is being maintained under lambda-cyhalothrin selection pressure in the laboratory in<br />
order to determine the potential for cross-resistance development.<br />
Results: Populations from Granada, Malaga, Valencia and Lerida showed a LC50 for lambdacyhalothrin<br />
of 144, 202, 134 and 287 ppm, respectively. The susceptibility of these populations<br />
is remarkably lower than that of a susceptible laboratory strain (LC50 = 18 ppm). In addition,<br />
laboratory selection with lambda-cyhalothrin initiated on a malathion-resistant laboratory strain<br />
has decreased its susceptibility to lambda-cyhalothrin from LC50= 98 ppm to LC50=759 ppm,<br />
whilst malathion susceptibility was maintained. Interestingly, the esterase inhibitor DEF<br />
partially suppressed the tolerance to lambda-cyhalotrin of the selected strain.<br />
Conclusions: The implications for Medfly control of the differences in susceptibility to<br />
lambda-cyhalothrin of field populations from different origins, when compared to laboratory<br />
strains, are discussed. Our results suggest that resistance to malathion may confer crossresistance<br />
to lambda-cyhalothrin.<br />
Keywords: Medfly, lambda-cyhalothrin, cross-resistance, malathion.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
SPINOSAD EFFICACY IN LABORATORY AND FIELD CONDITIONS AGAINST THE<br />
MEDITERRANEAN FRUIT FLY CERATITIS CAPITATA WIED. (DIPTERA:<br />
TEPHRITIDAE).<br />
Bachrouch, Olfa 1 ; Mediouni-Ben Jemâa, Jouda 2 ; Beitia, Francisco 3 & Chermiti,<br />
Brahim 4 .<br />
1 Unité de Recherche de Physico-Chimie Moléculaire. IPEST, BP51, 2070 La Marsa, Tunisia. 2 Laboratoire de<br />
Protection des Végétaux, Institut National de la Recherche Agronomique de Tunisie, 2049 Ariana, Tunisia.<br />
3 Associated Unit of Entomology, Centre of Plant Protection and Biotechnology, Instituto Valenciano de<br />
Investigaciones Agrarias (IVIA), Moncada (Valencia) SPAIN. 4 Institut Supérieur Agronomique de Chott-Mériem,<br />
Sousse, Tunisia.<br />
Background: In Tunisia, the Mediterranean fruit fly Ceratitis capitata, is a serious pest causing<br />
quantitative and qualitative losses to several crops, mainly citrus. The chemical control using<br />
broad-spectrum insecticides, essentially organophosphates, plays a major role in controlling<br />
this pest. However, insect resistance was recorded and control failures have been reported in<br />
field situations last years. Among the more used insecticides, malathion �diethyl (dimethoxythiophosphorylthio)<br />
succinate� has been declared an ozone-depleting substance and therefore,<br />
is being phased out. This work aims to evaluate Spinosad efficacy as an alternative to<br />
conventional chemical control in Tunisia.<br />
Methods: The assessment of Spinosad efficacy (dose=10ml/hl) was based on determination of<br />
adult and larval mortalities in laboratory together with population reduction expressed by<br />
weekly male captures and fruit damage at harvest in the field. Non treated insects were used as<br />
control in the laboratory experiment whereas in field conditions trials, a control plot of 0.5 ha<br />
area was employed.<br />
Results: Laboratory experiment showed that C. capitata larvae were more tolerant than adults.<br />
At 72h after treatment, Spinosad achieved 22% of mortality of larvae against 3% for the<br />
control, whereas, it leds to 100% mortality for adults against 4% for the control. For the field<br />
trial, results indicated significant differences in male captures and fruit damage percentages<br />
between Spinosad and the control. Concerning adult males captures reductions of 55.15% were<br />
recorded. For fruit damage assessment at harvest, results showed significant differences<br />
between the treated and the control fields. The percentage of punctured fruit was 13.13% in<br />
Spinosad trial against 20.40% for the control.<br />
Conclusions: Spinosad reduced significantly the damage produced by medfly females, and can<br />
be an efficient alternative to chemical control in Tunisia.<br />
Keywords: Ceratitis capitata, Spinosad, tolerance, fruit damage<br />
277
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
A COMPARISON OF SEVERAL TRAPS AND ATTRACTANTS AS A FIRST APPROACH<br />
TO THE USE OF MASS TRAPPING AGAINST THE OLIVE FRUIT FLY (DIPTERA,<br />
TEPHRITIDAE) IN THE VALENCIAN COMMUNITY.<br />
Beitia, Francisco J. 1 ; Tortosa, A. 2 ; Carbonell, Emilio 3 & Ros, J. Pedro. 4<br />
1 Associated Unit of Entomology, Centre of Plant Protection and Biotechnology, Instituto Valenciano de<br />
Investigaciones Agrarias (IVIA), Moncada (Valencia) SPAIN. Email: beitia_fra@gva.es. 2 Agricultural<br />
Experimental Station, IVIA, Carcaixent (Valencia) SPAIN. 3 Biometry Unit, IVIA, Moncada (Valencia) SPAIN.<br />
4 Instituto Nacional de Investigaciones Agrarias. Laboratorio de Entomología Agroforestal. Departamento de<br />
Protección Vegetal. Carretera de La Coruña, Km 7,5. Madrid, SPAIN.<br />
Background: The Valencian Community (on the eastern coast of Spain) is the fourth Spanish<br />
region in olive production (for oil and olives). One of the main problems for this crop is the<br />
olive fruit fly, Bactrocera oleae (Gmelin) and this pest has traditionally been combated in the<br />
region by the use of chemicals. Nowadays, it is been studied the technical and economical<br />
feasibility in using some ecological methods against the pest. One of these methods could be<br />
the mass trapping, so some trials have been started to know its effectiveness.<br />
Methods: The experiment was carried out in two olive plots located in Enguera (Valencia,<br />
Spain), an inner town about 70 km far from the Mediterranean coast. Two different olive<br />
varieties were in each plot: Arbequina and Villalonga. Five traps (glass MacPahil, Tephritrap,<br />
ecological Tephritrap, Easy trap and Olipe) and three attractants (Nulure+Borax, Tephri Lure<br />
and Ammonium Phosphate), with a total of 15 combinations (trap+attractant) were analyzed.<br />
Trials started on 5 th June and finished on 31 st December. Traps were installed on trees in both<br />
plots over an area of about 2.300 m 2 and every other tree. They were moved between trees<br />
through the experiment to allow all trap combinations were in all positions in the plot. Traps<br />
were moved, attractants renewed and insects inside traps counted every two weeks.<br />
Results: Data were similar in the two plots, so in the two olive varieties. The best trap and the<br />
best attractant in capturing olive flies were Tephritrap and Tephri Lure respectively. In<br />
addition, the best combination was Tephritrap + Tephri Lure. Ammonium Phosphate shows the<br />
worst result in capturing flies, and this compound with Easy Trap and Olipe were the worst<br />
combinations. In general, all combinations in both plots show two period of high fly captures:<br />
through the month of September and on mid October. In the other hand, captures of natural<br />
enemies in traps were also counted and specially for Chrysopa sp individuals, a very common<br />
and important natural enemy on olive groves. The best trap in capturing flies was also one of<br />
the traps with a low ratio nº of chrysopas/nº of flies (0’07 in both plots) and these data must be<br />
considered to determine the real effectiveness of the traps.<br />
Conclusions: Mass trapping could be an interesting method to be applied against the olive fruit<br />
fly in the Valencian Community but it must be analyzed not only the effectiveness of the trap<br />
but also its cost and its effect on natural enemies in order to select the best one.<br />
Keywords: Olive fruit fly, traps and attractants, mass trapping.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
LABORATORY EVALUATION OF THE EFFECTS OF ATTRACT & KILL<br />
FORMULATIONS ON ANASTREPHA FRATERCULUS.<br />
Borges, Rafael 1 ; Botton, Marcos 2 ; Machota Jr., Ruben 2 ; Boff, Mari Ines Carissimi 3 ;<br />
Mafra-Neto, Agenor 4<br />
1 ISCA Tecnologias, BR 285, Km 461, Ijui, RS 98700 Brasil. Email: rafael@isca.com.br. 2 Empresa Brasileira de<br />
Pesquisa Agropecuária (EMBRAPA-CNPUV). Bento-Gonçalves, RS (Brasil). 3 Universidade do Estado de Santa<br />
Catarina, Centro Agroveterinário, Lages, Brasil. 4 ISCA Technologies, Riverside California, US.<br />
Background: A new atract & kill (A&K) formulation was developed by ISCA Tecnologias to<br />
control fruit flies. In laboratory tests, we compared the efficacy of this new formulation with<br />
three commercial products used by fruit growers in southern Brazil. Mortality tests were<br />
performed with Anastrepha fraterculus (Diptera: Tephritidae) and results showed that the<br />
effectiveness of Anarosa formulations was directly proportional to increasing dose of<br />
insecticide. Formulations containing organophosphates and cypermethrin produced similar<br />
mortality results. The Anarosa formulations, regardless of the insecticide dose, were more<br />
effective than GF 120 for A. fraterculus control.<br />
Methods: Experiments were conducted at the Entomology Laboratory of EMBRAPA Grapes &<br />
Wine, Bento Gon‡alves, RS (temperature 25øCñ2, relative humidity (RH%) 70ñ10,<br />
photoperiod 14 hours) with insects reared on a diet of wheat germ, brown sugar and soy extract<br />
(3:1:1). Mortality of adult 15-20 days of age A. fraterculus was evaluated. Each replicate<br />
consisted of a cage (300ml plastic cup without bottom, containing a circular floor paper, a 2 ml<br />
container for water and 0.8g of solid diet) with two pairs of adult insects. Treatments were<br />
applied to leaves of Vitis vinifera plants in a greenhouse setting. Using disposable syringes, a<br />
5ml drop (4 mm in diameter) was placed on the upper leaf surface. After a three hour drying<br />
period, the leaves were collected in the laboratory and cut into a circular shape (3cm diameter)<br />
containing a drop of the treatment which was then presented to the adult flies. Each treatment<br />
was replicated 13 times; each cage was a replicate. Measurements were taken every two hours<br />
for the first 24 hours, followed by intervals of four hours until the completion of assessments<br />
after 96 hours.<br />
Results: The most rapid mortality resulted from the Anarosa + 0.2% cypermethrin and Biofruit<br />
+ 0.15% malathion treatments, which caused 100% mortality after 36 and 32 hours<br />
respectively. Anarosa + 0.4% spinosad resulted in 100% mortality after 44 hours. The Anarosa<br />
+ 0.2% spinosad and molasses + 0.15% malathion treatments both resulted in 100% mortality<br />
after 48 hours. For the Anarosa treatments containing 0.1% and 0.05% spinosad, 100%<br />
mortality was reached after 52 and 60 hours for each treatment respectively. The GF 120<br />
treatment resulted in 96% mortality at the end of the tests, while the standard treatment,<br />
Anarosa without insecticide, resulted in 13% mortality at the end of the experiment.<br />
Conclusion: Anarosa treatments proved to be promising for controlling populations of<br />
Anastrepha fraterculus and the mortality effect of the formulations progressively increases with<br />
increasing insecticide concentration<br />
Keywords: Anarosa, Spinosad, Mortality, Anastrapha fraterculus<br />
279
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PERSISTENCE OF ATTRACT & KILL FORMULATIONS OF ANASTREPHA<br />
FRATERCULUS: GREENHOUSE AND RAIN-FAST EXPERIMENTS.<br />
Borges, Rafael 1 ; Botton, Marcos 2 ; Machota Jr., Ruben 2 ; Boff, Mari Ines Carissimi 3 ;<br />
Bortoli, Ligia Caroline 4 ; Mafra-Neto, Agenor 5 .<br />
1 ISCA Tecnologias, BR-285, Km-461, Ijui, RS 98700 Brasil, rafael@isca.com.br. 2 Empresa Brasileira Pesquisa<br />
Agropecuária-CNPUV, RS-Brasil. 3 Universidade Estado Santa Catarina, Centro Agroveterinário SC-Brasil.<br />
4 Universidade Caxias Vinhedos, RS-Brasil; 5 ISCA Technologies, Riverside California, US.<br />
Background: Food attractants associated with pesticides have been used as a strategy for<br />
management of fruit flies in fruit growing areas of Southern Brazil. Commercial formulations<br />
require frequent applications due to environmental factors like sunlight and rain. Anarosa,<br />
developed by ISCA Technologies, was evaluated for persistence after exposure to solar<br />
radiation and rainfall in laboratory tests on mortality of adult Anatrepha fraterculus.<br />
Material and Methods: Experiments were conducted to evaluate the persistence of two attract<br />
and kill (A&K) formulations of Anarosa containing 0.1 or 0.2% Spinosad, compared to two<br />
standard treatments, GF 120 (0.02% Success® CB) and hydrolyzed protein (Biofruit®) +<br />
malathion on adult A. fraterculus. Experiments were conducted on Vitis vinifera in a<br />
greenhouse setting. Five plants were used per treatment. Treatments were applied as droplets<br />
(4mm diameter, 20 droplets) to upper leaves using disposable syringes (5ml). After 1, 7, 14,<br />
21, 28 and 35 days after application, leaves with treatments were collected for rain-fast testing.<br />
Each treatment was exposed to 20 and 50mm of artificial rain; then leaf samples were collected<br />
and presented to flies in the laboratory to measure mortality. Each treatment was composed of<br />
13 replicates consisting of a cage (300ml plastic cup without bottom, containing a circular floor<br />
paper, a 2ml container for water and 0.8g of solid diet) with two pairs of adult insects. For each<br />
evaluation, leaves with treatment droplets were cut in a circular shape (3cm diameter) and<br />
presented to the insects. Measurements were taken every two hours for the first 24 hours,<br />
followed by every four hours until the completion of assessments after 96 hours.<br />
Results: Samples exposed to 20 mm of simulated rainfall resulted in 75 and 83% mortality<br />
after 96 hours for Anarosa 0.1 and 0.2% Spinosad treatments, respectively whereas GF 120 +<br />
malathion and Biofruit + malathion each resulted in only 39 and 18% mortality after 96 hours.<br />
The control sample resulted in 4% mortality. Samples exposed to 50 mm of simulated rainfall<br />
resulted in 59 and 83% mortality for Anarosa 0.1 and 0.2% spinosad treatments, 9 and 4% for<br />
the GF 120 + malathion and Biofruit + malathion treatments and 4% for the control treatment.<br />
Samples exposed to solar radiation showed no significant differences in mortality after 1 to 21<br />
days with all resulting in 100% mortality.<br />
Conclusions: The persistence of both the Anarosa and GF 120 formulations was similar after<br />
exposure to solar radiation, however the Anarosa formulation proved highly persistent with<br />
respect to rain, maintaining efficiency after considerable rainfall volumes.<br />
Keywords: South American fruit fly, rain-fast, solar radiation, Anarosa, GF 120
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FRUIT INFESTATION ON SOME CUCURBITS AS INFLUENCED BY PUPAL DENSITY OF<br />
MELON FLY, BACTOCERA CUCURBITAE (COQUILLETT) IN SOIL AND THEIR<br />
MANAGEMENT THROUGH SOIL TREATMENTS.<br />
Chatterjee, Hirak 1 & Laskar, Nripendra 2<br />
1 Department of Plant Protection, Palli Siksha Bhavana, Visva-Bharati, Sriniketan-731236, India.Email:<br />
hc65@rediffmail.com. 2 Department of Agricultural Entomology, Uttar Banga Krishi Viswavidyalaya,<br />
Coochbehar, Pundibari-736165, India.<br />
Background: Melon fly, Bactocera cucurbitae (Coquillett) is the most notorious pest of fruit<br />
vegetables causing considerable damage worldwide. This biotic pressure is increasing day by<br />
day due to intensive cultivation of HYVs under chemicalized agriculture, which leads to<br />
several environmental ill effects. Therefore, a sustainable management technology is a dire<br />
need of the hour. Here a novel eco-friendly approach has been made to combat this pest<br />
thorough estimation of pupal density in soil, its impact on percent fruit infestation and<br />
evaluation of some pesticide molecules including microbial.<br />
Methods: Four important cucurbitaceous vegetables in West Bengal, India viz. pumpkin,<br />
Cucurbita moschata L., bittergourd, Momordica charantia L., ridgegourd, Luffa acutangula<br />
Roxb. and cucumber, Cucumis sativus L. were selected and the fruits were observed for melon<br />
fly infestation at weekly interval. Subsequently from each field soil samples below 12 cm<br />
depth were collected for pupal count. For evaluation of residual toxicity of some chemicals,<br />
soil was collected from experimental plot and after oven drying the same was kept in a plastic<br />
container followed by insecticide application. The third instar maggots at ‘jumping stage’ were<br />
released in the test container after 1, 3, 5 and 7 days of application of insecticides. The data<br />
thus obtained on various stages of development were subjected to statistical analysis by<br />
appropriate techniques.<br />
Results: Highly significant positive correlations were established between pupal density and<br />
fruit infestation with having correlation coefficient ‘r’ = 0.984, 0.987, 0.962 and 0.923 on<br />
pumpkin (Cucurbita moscheta L.), bittergourd (Momordica charantia L.), ridgegourd (Luffa<br />
acutangulata Roxb.) and cucumber (Cucumis sativus L.) respectively. Economic threshold<br />
level (ETL) worked out from the individual regression model showed 0.47, 0.24, 0.53 and 0.56<br />
adult male per trap per day for the respective cucurbit vegetables. Among the different<br />
chemicals significant differences in pupal mortality, pupal deformity and adult emergence were<br />
observed. Soil application of dichlorvos 76 EC emerged as most effective with highest mean<br />
pupal mortality (63.33%) and lowest mean adult emergence (10%) followed by carbofuran 3G<br />
( 58.33, 12.17%) and chlorpyriphos 20EC (56.25, 15.42%). But bleaching powder and<br />
formaldehyde (4%) proved to have insignificant effect on adult eclosion as 80.33 and 76.33%<br />
adult already eclosed from bleaching powder and formaldehyde treated soil respectively.<br />
Whereas maximum deformed pupae ( 28.75%) was recorded from the soil treated with the<br />
microbial biopesticide, Metarhizium anisopliae (1.15WP).<br />
Conclusions: The above findings established that the percent fruit infestation is highly<br />
influenced by its pupal density in field soil and their different developmental stages could be<br />
disrupted through soil application of chemicals. Also suggest that it could be an efficient tool<br />
for sustainable management of melon fruit fly before their emergence.<br />
Keywords:, Bactocera cucurbitae, fruit infestation, pupal density, mortality, deformity, eclosion, soil treatments<br />
281
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
OLIVE FRUIT FLY, BACTROCERA OLEAE GMEL., MASS-TRAPPING WITH OLIPE<br />
TRAPS: EFFECT OF HOLE SIZE IN THE FLY AND NON-TARGET ARTHROPODS<br />
CAPTURES.<br />
Coelho, Valentim 1* ; Bento, Albino A. 1 ; Mexia, A. 2 ; Pereira, José A. 1<br />
1 CIMO/Escola Superior Agrária, Instituto Politécnico de Bragança, Campus Sta Apolónia, Apt.<br />
11172, 5301-855 Bragança, Portugal; Email: jpereira@ipb.pt; 2 Instituto Superior de<br />
Agronomia, Tapada da Ajuda, 1349-017 Lisboa, Portugal<br />
Background: The olive fruit fly, Bactrocera oleae Gmelin (Diptera: Tephritidae), is a key-pest<br />
of olive tree in the Mediterranean region. In organic agriculture, the use of Olipe traps for olive<br />
fruit fly mass-trapping is currently used in different regions of the Iberian Peninsula. However,<br />
previous works have demonstrated that trap hole size influences the number of fly and nontarget<br />
arthropods captures, that could have a significant impact of beneficial arthropods<br />
communities. In this context, the present work aimed to study these aspects as well as the<br />
efficacy of the method in the control of the pest.<br />
Methods: The work was conducted in an organic olive grove located in Mirandela (Northeast<br />
of Portugal). The olive trees have medium size and belong to Cobrançosa variety. Four<br />
treatments (bottle hole sizes) were selected (4, 6, 8 and 10 mm), and each tested in one hectare<br />
orchard at basis of one trap per tree. In each treatment with a bi-weekly basis 15 traps were<br />
changed and the number of flies and no-target arthropods were counted. The attack of fruits<br />
was observed in 25 fruits per tree in a total of 20 trees.<br />
Results: In all treatments a total of 8922 individuals were counted and ants were the major<br />
group recovered. A marked effect was observed in the total number of non-target arthropods if<br />
ants are excluded. The use of these bottles was particularly harmful to the adults of the<br />
Chrysopidae family that represents between 3.7% and 17.6% of the total recovered (excluding<br />
ants). Concerning to B. oleae, a reduced number of adults were observed in all treatments. No<br />
differences were registered in the attacked fruits between bottle hole size. And, the fruit attack<br />
reaching 19% in the control plot (without traps) that was about the double of treated plots.<br />
Keywords: Olive fruit fly, Olipe traps, hole size, no target arthropods, organic agriculture
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
MEDFLY CONTROL IN MEDITERRANEAN AREAS OF SPAIN WITH M3 TRAPS.<br />
Coltell, Néstor * ; Espinosa, Eduardo & Sabater Alepuz, Juan B.<br />
BIAGRO SL. Poligono industrial del Mediterráneo. Jaime I, n8. 46560 Massalfasar, Valencia (Spain). Email:<br />
biagro@biagro.es.<br />
Background: The Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), is a keypest<br />
of fruit trees in the Mediterranean region. Lure and kill traps as M3 represent an efficient<br />
system to control fruit flies. Several trials are presented in this work to ascertain its efficiency<br />
under different mediterranean conditions and in different crops.<br />
Methods: The work was conducted in several citrus and stone fruits orchards Bétera, Vilavella,<br />
Sagunto, Pobla de Farnals, Alama de Murcia, Jumilla, Yecla and Lérida. Traps were hold in<br />
place at 360 traps per ha, at 45 days prior fruit harvest. Fruit damage was evaluated in all<br />
orchards. Control traps baited with food attractants were placed in the orchard perimeter and<br />
one in the center.<br />
Results: In all the assays a significant fruit damage reduction has been obtained when<br />
compared to control orchards subjected to chemical control. Its broad application for organic<br />
agriculture is discussed.<br />
Keywords: M3 traps, fruit damage, organic agriculture<br />
283
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECTIVE SAMPLING RANGE FOR PROTEIN-BASED LURES.<br />
Epsky, Nancy D.* 1 ; Kendra, Paul E. 1 ; Espinoza, Hernán R. 2 ; Midgarden, David 3 &<br />
Heath, Robert R. 1<br />
1 United States Department of Agriculture, Agricultural Research Service (USDA, ARS), Subtropical Horticulture<br />
Research Station, 13601 Old Cutler Road, Miami, Florida, 33158, USA;. Email: nancy.epsky@ars.usda.gov;<br />
2 Fundación Hondureña de Investigación Agrícola (FHIA), La Lima, Cortés, Honduras; 3 United States Department<br />
of Agriculture, Animal and Plant Health Inspection Service - International Services (USDA, APHIS-IS), Sao<br />
Paulo, Brazil<br />
Background: Effective sampling range is defined as the maximum distance from which an<br />
insect can reach an attractive source in a given period of time. Information on sampling range<br />
is valuable for determining appropriate coverage of traps for use in population delimitation,<br />
mass trapping control strategies, or identification of foci of infestation for precision targeting of<br />
control measures. Despite extensive use of protein-based female-targeted attractants in current<br />
trapping programs for tropical tephritids, there is little documentation their effective sampling<br />
range.<br />
Methods: Studies were conducted in Florida and Honduras. A geostatistical analysis approach,<br />
using data from capture of wild flies in a specifically designed trapping grid, was compared<br />
with a standard release/recapture study approach that used laboratory-reared sterile and wild<br />
flies (Florida), and recently captured wild flies (Honduras) to determine effective sampling<br />
range. Field design consisted of 38 traps (over 0.5 ha) placed in a combination of standard and<br />
high density grids to facilitate geostatistical analysis, and tests were conducted in coffee,<br />
mango and orthanique with the three component lure (APT: ammonium acetate, putrescine,<br />
trimethylamine, Honduras) and guava with the two component lure (AP) and torula yeast/borax<br />
(TY, Florida).<br />
Results: Effective sampling range, as determined by geostatistical analysis for APT for C.<br />
capitata, was ~30 m for flies captured in coffee or mango and ~ 40 m for flies captured in<br />
orthanique. Similar results were obtained from the release/recapture study, and contour maps<br />
indicated that wind direction had a strong influence on sampling range, which was ~15 m<br />
greater upwind as compared to downwind from the release point. In release/recapture tests with<br />
mature wild A. suspensa, range of AP was determined to be 30 m. With sterile females,<br />
effective range of both AP and TYB was 20 m. Sterile immature A. suspensa females dispersed<br />
farther and were recovered in higher numbers than mature females, and recapture with AP was<br />
higher than recapture with TY.<br />
Conclusions: Geostatistical analysis of field-captured insects in appropriately designed<br />
trapping grids may provide a supplement or alternative to release/recapture studies to estimate<br />
sampling ranges for semiochemical-based trapping systems. Contour maps indicated that wind<br />
direction had a strong influence on the active space of attractants, as reflected by distribution of<br />
captured flies in release/recapture studies.<br />
Keywords: Ceratitis capitata, Anastrepha suspensa, synthetic lures, torula yeast
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FIELD ASSAYS OF NEW CONTROLLED-RELEASE PHEROMONE DISPENSERS FOR<br />
CONTROLLING BACTROCERA OLEAE (GMELIN).<br />
Gil-Ortiz, Ricardo*; Moya-Sanz, Pilar 1 ; Navarro-Fuertes, Ismael 1 ; Navarro-Llopis,<br />
Vicente 1 & Primo, Jaime 1<br />
1 Centro de Ecología Química Agrícola (CEQA), Universidad Politécnica de Valencia. Camino de Vera s/n.<br />
Edificio 6C. 46022 – Valencia (Spain). Email: mmoyasa@ceqa.upv.es<br />
Background: The eco-trap systems for controlling the olive fly, Bactrocera oleae (Diptera:<br />
Tephritidae), are used worldwide in integrated pest management programmes. The low<br />
biodegradability of most commercial pheromone dispensers requires finding other alternatives<br />
more ecological to the currently existing ones. We report here on several type of eco-friendly,<br />
hand-applied pheromone (1.7-dioxaspiro [5.5] undecane) dispensers tested in both laboratory<br />
and field.<br />
Methods: Clay materials and different additives, in changing proportions, were designed and<br />
evaluated in order to obtain that possessing the optimal pheromone emission. The effectiveness<br />
of the best formulations were evaluated in field and subjected in parallel to a procedure of<br />
accelerated aging in a temperature and wind speed controlled chamber. Residual pheromone<br />
remaining in the dispensers was periodically evaluated by gas-liquid chromatography. Release<br />
rates and half-life times were determined.<br />
Results: Two formulations (four types of dispensers) showed the best performances in the<br />
laboratory as compared with the commercial B. oleae Long Life Lure from Agrisense ®<br />
(Pontypridd, UK) so they were finally tested in the field. Design 1E Type showed the best<br />
behaviour in the field with an average controlled release of emission of 0.6 mg<br />
pheromone/dispenser.day, from day 21, which is slightly higher than that showed by the<br />
commercial one. In addition, it showed an initial loss of pheromone 10% lower than that<br />
showed by Long Life Lure. Maximum differences in emission rates of all dispensers ranged<br />
between 0.1-0.9 mg pheromone/day. The average of captures obtained in SEVEP ® traps (EPA,<br />
SP) were not statistically significant despite the differences in emission found among<br />
dispensers. The best half-life time obtained was higher than 4 months, which allows to<br />
dispenser to be placed in the field without replacement during an entire olive fly campaign in<br />
the Mediterranean area of Spain.<br />
Conclusions: The field emission tests of dispensers designed confirmed the good performance<br />
obtained in the laboratory. The mean of captures were as effective as the best dispenser on the<br />
market, which combined with their easy biodegradability make to be suitable for monitoring<br />
and controlling B. oleae, especially in "Lure and kill" programs.<br />
Keywords: Olive fly, dispenser, pheromone, monitoring, control.<br />
285
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
STRUCTURAL ABNORMALITIES IN THE EGGS OF FRUIT FLY, BACTROCERA TAU<br />
(WALKER) AFTER FEEDING ON PHLOROGLUCINOL.<br />
Gupta, Divender* & Sunandita<br />
Department of Entomology and Apiculture. Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni<br />
(Solan)-HP-173 230 India.<br />
Background: Fruit fly, Bactrocera tau is a major pest of tomato and cucurbit vegetables and<br />
loads of insecticides are used to prevent the damage by this fly but in the process the fruit gets<br />
poisoned making the consumption unsafe. Phloroglucinol (1,3,5 trihydroxy benzene) has been<br />
reported as an egg shell peroxidase inhibitor and here in the present study we report the effect<br />
of this novel chemical on the eggs of B. tau when fed on artificial diet containing this chemical,<br />
under laboratory conditions.<br />
Methods: In the first set of experiment, five pairs of freshly emerged adults of B. tau were<br />
given a diet containing glucose, protein hydrolysate and water (4:1:5) provided with different<br />
concentrations of phloroglucinol (1,5 and 10mM (milli Molar) in first subset, and 25, 50 and<br />
100 mM in second subset). These fruit flies were provided with a host (cucumber) for egg<br />
laying from 9 th day onwards. In the second set of experiment, 8 day old flies were provided<br />
with phloroglucinol containing diet and the host was provide for egg laying as in the first set.<br />
Observations on egg laying and the structure of eggs were taken.<br />
Results: In Set-1, where the adults were fed continuously on phloroglucinol containing diet,<br />
normal egg laying was recorded at 1, 5 and 10mM phloroglucinol concentrations. Egg count<br />
was slightly less (158.0eggs/female) at 25mM concentration in comparison to 187.0<br />
eggs/female in control. However at 50 and 100mM concentrations of phloroglucinol the egg<br />
laying reduced drastically to 26.2 and 12.0 eggs/female, respectively. Further, the eggs laid<br />
were fragile, very thin or with ruptured chorion at 50mM, whereas at 100mM in addition some<br />
chorionless eggs or vitelline membrane bound oocytes (VMOs) were also laid. In Set-II where<br />
the diet containing phloroglucinol was provided after 8 days of emergence, a similar trend was<br />
recorded.<br />
Conclusions: Phloroglucinol has a marked effect on egg development of B. tau particularly on<br />
egg chorion as it acted by inhibiting egg shell peroxidase when fed at 50 and 100mM<br />
concentrations. It resulted in eggs with thin chorion, ruptured chorion and vitelline membrane<br />
bound oocytes (VMOs), which failed to hatch. The study suggests that it can be an effective<br />
ecofriendly tool in IPM of B. tau.<br />
Keywords: Fruit fly, Bactrocera tau, phloroglucinol
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
FRUIT FLIES SPECIES COMPOSITION AND SEASONALITY, AND RELATIVE<br />
EFFICIENY OF FOUR FEMALE ATTRACTANTS IN MANGO ORCHARDS IN BENIN.<br />
Hanna, Rachid 1* ; Gnanvossou, Desiré 1 ; Bokonon-Ganta, Aimé 1 & Ekesi, Sunday 2<br />
1 International Institute of Tropical Agriculture (IITA), 08 BP 0932 Tri Postal, Cotonou Benin. Email:<br />
r.hanna@cgiar.org; 2 International Centre for Insect Physiology and Ecology (icipe), Nairobi, Kenya.<br />
Background: Trapping of tephritid fruit flies has received a great deal of attention for<br />
detection, monitoring and control purposes. Several commercially available female attractants<br />
have been developed and are widely used, but information on their usefulness in fruit fly<br />
monitoring under various environments in Africa is scarce. Here we report on species<br />
composition and seasonality of fruit flies caught in four commercial female attractants in<br />
mango orchards in Benin (West Africa).<br />
Methods: The experiments were conducted in eight mango orchards in 2007 and 2008 and 12<br />
orchards in 2009 – spread across four agroecologies in Benin. Orchards contained a range of<br />
mango cultivars representing what is grown in the targeted areas. Fruit fly trapping was carried<br />
out each year during the mango fruiting period – March through July. Four commercially<br />
available attractants were tested using Multilure traps. Three traps per attractant and per<br />
orchard were used. Torula yeast was used as the standard in all three years and was compared<br />
with Nulure in 2007, BioLure in 2008, and Mazoferm in 2009. Traps were serviced at weekly<br />
and biweekly intervals for liquid and dry attractants, respectively.<br />
Results: Overall, eleven fruit fly species belonging to three genera were found in traps, with<br />
Bactrocera invadens and Ceratitis cosyra being the most abundant. Species composition in<br />
traps mirrored those in infested mango fruits, with species relative abundance depending on<br />
agroecology. Bactrocera invadens was present in all study sites but its seasonality followed a<br />
pattern of dominance throughout the mango season in the south while being nearly absent in<br />
the drier agroecologies in northern Benin during the first half of the season, but its abundance<br />
increased steadily in central and northern Benin with the onset of rains and became the<br />
dominant species during the second half of the season. Ceratitis cosyra abundance followed<br />
patterns opposite to those of B. invadens. Torula yeast attracted considerably more fruit flies<br />
than BioLure (20% of Torula yeast) regardless of agroecology (P < 0.05), but attracted similar<br />
numbers of flies as Mazoferm, except in the Sudan Savanna where Torula yeast attracted more<br />
B. invadens (P = 0.008) and Ceratitis spp. (P = 0.048) than Mazoferm. Nulure was generally<br />
less attractive to all fruit flies than Torula yeast.<br />
Conclusions: The study revealed that Torula yeast and Mazoferm are equally attractive to the<br />
fruit fly species assemblage in mango orchards in Benin, followed by Nulure and BioLure.<br />
There were not any significant differences; however, in relative attractiveness of the four<br />
attractants to B. invadens and Ceratitis spp. Implications for fruit fly monitoring and<br />
suppression are discussed.<br />
Keywords: Multilure traps, protein bait, Torula yeast, Nulure, BioLure, Mazoferm.<br />
287
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ORGANOPHOSPHATE RESISTANCE IN ORIENTAL FRUIT FLY MAY AFFECT THE<br />
ATTRACTING EFFICIENCY OF OP-BASED INTOXICANT LURE.<br />
Hsu, Ju-Chun 1* ; Hai-Tung Feng 2<br />
1 Department of Entomology, National Taiwan University, Taipei, Taiwan. Email: juchun@ntu.edu.tw; 2 Taiwan<br />
Agricultural Chemicals and Toxic Substances Research Institute, Council of Agriculture, Taiwan<br />
Background: The oriental fruit fly (Bactrocera dorsalis (Hendel)) is an economic pest of<br />
great significance in the Asia-Pacific region. The most common management tactic is the male<br />
annihilation technique (MAT), which consists of using highly attractive toxic lures. The<br />
toxicant used in these lures are most often organophosphate (OP) based insecticides. In recent<br />
years, fruit flies in Taiwan have been reported to have OP resistance in the field. Similar cases<br />
of resistance development and subsequent effectiveness reductions to OP based insecticides<br />
have been observed in a wide range of alternate fly species in different localities. However, at<br />
this time, there have been no evaluations concerning the control efficiency of OP-based<br />
intoxicant lures when resistance is exhibited in the field.<br />
Methods: The attracting efficiency of naled with methyl eugenol (ME) to two different levels<br />
of OP-resistant and one susceptible male oriental fruit fly lines were evaluated under<br />
greenhouse conditions in Taiwan. Two concentrations of naled (5 and 2.5%) with 90% of ME<br />
were used to evaluate the attraction efficiency within the traps, or cages, after 1 h, 24 and 48 h<br />
post-treatment. Flies from each of the 3 lines were placed inside a perforated plastic fruit fly<br />
trap, and then the traps were suspended by means of a wire attached to the roof of a organdy 1<br />
m 3 cage.<br />
Results: The attracting efficiency of B. dorsalis males in the three lines were higher for the 5%<br />
naled treatments. Of the three lines, attraction was highest in the higher OP-resistant line,<br />
followed by the lower OP-resistant line, and the least efficiency was exhibited in the<br />
susceptible line at different posttreatments times.<br />
Conclusion: In our study, OP-resistant lines affected the attracting efficiency of OP-based<br />
intoxicant lures. When applying MAT to oriental fruit fly control, the intoxicant resistance<br />
needs to be considered.<br />
Keywords: Bactrocera dorsalis, MAT, naled, methyl eugenol.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
IMPROVED FRUIT FLY LURES AND FORMULATIONS FOR DETECTION,<br />
DELIMITATION AND CONTROL.<br />
Jang, Eric * 1 ; Vargas, Roger, 1 ; Ramsey, Amanda 2 , Khriman, Ashot 3 , & Siderhurst,<br />
Matthew 4 .<br />
1 USDA-ARS, Pacific Basin Agricultural Research Center, POB 4459, Hilo, HI 96720 USA Emmail:<br />
eric.jang@ars.usda.gov 2 Scentry Biologicals, 610 Central Avenue, Billings, MT 59102, USA 3 USDA-ARS,<br />
Invasive Insect Biocontrol and Behavior Laboratory, Bldg 007, BARC-West, Beltsville, MD, 20705, USA, 4<br />
Eastern Mennonite University, 1200 Park Road, Harrisonburg, VA, 22802, USA<br />
Background: Exotic fruit fly detection, delimitation and control using semiochemicals-based<br />
attractants form the foundation of area-wide pest management programs worldwide.<br />
Increasingly, countries require detection arrays as a basis for certification of fly-free areas, low<br />
prevalence areas and for development of systems approaches which all facilitate international<br />
trade. Improved attractants and formulations for key fruit fly pests such as Ceratitis sp.,<br />
Anastrepha sp., as well as methyl eugenol and cue-lure responding Bactrocera are needed to<br />
improve detection, reduce costs and facilitate auditing of regulatory areas of concern. We<br />
report here on recent developments to improve lures and formulations for use in fruit fly<br />
programs.<br />
Methods: New (improved) attractants have been developed for Ceratitis (ceralure B1), methyl<br />
eugenol-responding Bactrocera (Iodo-eugenol) and cue-lure responding Bactrocera (melolure<br />
and cuc-lure). Attractants were identified, formulated and tested in the field against<br />
representative species and in some cases multiple species in conjunction with collaborators.<br />
Data were collected in a variety of bioassay arenas including flight tunnels, field cages and in<br />
open field tests lasting up to 10 weeks or more. Additionally, several new and/or improved<br />
formulations of existing attractant lures have been developed that reduce handling of liquid<br />
lures on cotton wicks and include toxicants in the formulations.<br />
Results: Many of the new attractants were shown to me more attractive than their predecessors<br />
based on molecule to molecule comparisons (ceralure B1 or melolure), or showed increased<br />
attractancy and female bias relative to proteinaceous baits (cuc-lure). These new attractants<br />
have also been tested in newer formulations such as solid matrix composites that allow the lure<br />
to volatilize at a more constant rate than the cotton wicks. Of the various toxicants tested,<br />
incorporation of naled or DDVP into the solid matrix as been effective, while reduced-risk<br />
insecticides such as spinosad or rynoxypur have has success with metyl eugenol containing<br />
formulations but less so with cue-lure formulations under field conditions.<br />
Conclusions: New and improved lures and formulations of fruit fly attractants have improved<br />
usability and detection capabilities for several tephritid species. These formulations should be<br />
considered by action agencies and regulatory bodies for inclusion into standardized detection<br />
grids or for control technologies such as mass-trapping, male annilhilation and bait sprays.<br />
Keywords: Tephritidae, Bactocera, Ceratitis, trapping, detection, control.<br />
289
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
IMPACT OF THE PROJECT ON INTEGRATED MANAGEMENT OF FRUIT FLIES IN<br />
INDIA (IMFFI) IN FRUITS AND VEGETABLES CULTIVATION IN SOUTHERN<br />
KERALA.<br />
Jiji, T. 1 , Verghese, Abraham 3 ; Suja, G. 3 , Naseema, A 4 ; Nisha, V.G; 5 Sarika, Mohan 6 &<br />
Mumford, John M. *<br />
1 Department of Agricultural Entomology, College of Agriculture, Vellayani, Thiruvanathapuram 695522, Kerala,<br />
India. 2 Indian Institute of Horticultural Research, Bangalore, India. 3 Regional Agricultural Research Station,<br />
Kayamkulam, India. 4, 5. College of Agriculture, Vellayani, Thiruvanathapuram, Kerala, India. 6 Imperial College<br />
London, UK.<br />
Background: Oriental fruit fly,Bactrocera dorsalis Hendel and melon fly B. cucurbitae Coq.<br />
are serious pests of mango and cucurbits, respectively. B. dorsalis is a direct pest on mango. In<br />
India, the loss in fruit yield ranges from 1 to 31% (Verghese et al., 2002). In vegetables the<br />
extent of crop loss due to B.cucurbitae varies between 30 to100 % (Sookar et al., 2006). The<br />
adult female lays eggs in maturing fruits and vegetables. The maggot develops in the fruit,<br />
causing drop and rotting. ICAR–UKDFID project on integrated management of fruit flies in<br />
India (IMFFI),which was in operation during 2002-2005 came out with packages for fruit fly<br />
management in India both for B. dorsalis and B. cucurbitae. The objective of the present<br />
study was to assess the impact of the research results of IMFFI in reducing the crop loss due to<br />
fruit fly attack in mango and cucurbìtaceous vegetables in southern Kerala [south India]. The<br />
study also assessed the feedback of farmers on the effectiveness of this technology and the<br />
scope of its commercial adoption.<br />
Methods: Trials in three southern districts of Kerala were carried out in farmers’ fields,<br />
maintaining demonstration plots, during April-June, 2009 to evaluate the efficacy of integrated<br />
pest management involving MAT using methyl eugenol traps/cue lure traps and Bait<br />
application technique (BAT), using food baits for the management of fruit flies infesting<br />
mango and snake gourd. Field sanitation and biocontrol methods were also integrated.<br />
Adoption of the technology was also assessed, based on feed back.<br />
Results: There was significant reduction in the level of B. dorsalis infestation in farmers’ fields<br />
where IPM was followed. In mango, in the three districts of Thiruvananathapuram, Kollam and<br />
Alappuzha the control plots recorded an infestation level of 80.41, 80.89 and 83.38 % as<br />
against the treated plots with a pest incidence of 8.41, 7.6 and 8.24 %, respectively. In<br />
snakegourd in the three districts Thiruvananthapuram, Kollam and Alappuzha, the control plots<br />
recorded an infestation level of 73.74, 73.16 and 74% as against the treated plots with a pest<br />
incidence of 7.4, 7.77 and 4.99%, respectively. The results are in conformity with the finding<br />
of the ICAR-UK DFID project on fruit flies. Beauveria bassiana owing to high infectivity and<br />
easiness of application gave encouraging results for the management of fruit flies..<br />
Conclusions: The application of biocontrol agent can effectively be incorporated as a<br />
component of integrated fruit fly management programme in mango and vegetable crops. The<br />
feedback was encouraging, which authenticates the findings of the IMFFI Project<br />
Keywords: IPM, IMFFI, B. dorsalis , B. cucurbitae, snake gourd.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
SIDE EFFECTS OF SELECTED PESTICIDES COMMONLY USED IN CITRUS ORCHARDS<br />
ON STERILE MEDFLY MALES.<br />
Juan-Blasco, María 1* ; Planes, Laura 1 ; Argilés, Rafael 2 ; Jacas, Josep A. 3 ; Sabater-Muñoz,<br />
Beatriz 1 & Urbaneja, Alberto 1 .<br />
1 Unidad Asociada de Entomología UJI-IVIA-CIB CSIC. Centro de Protección Vegetal y Biotecnología; Instituto<br />
Valenciano de Investigaciones Agrarias, IVIA; Ctra. de Moncada a Náquera km 4.5; E-46113-Moncada (Spain).<br />
Email: juan_marbla@gva.es. 2 Departamento de Plagas. TRAGSA. Release Unit. Ctra. de Moncada a Náquera km<br />
4.5; E-46113-Moncada (Spain). 3 Unidad Asociada de Entomología UJI-IVIA-CIB CSIC. Departament de<br />
Ciències Agràries i del Medi Natural; Universitat Jaume I, UJI; Campus del Riu Sec; E-12071-Castelló de la<br />
Plana (Spain).<br />
Background: The Sterile Insect Technique (SIT), can be considered a biological control<br />
method, as it is based on rearing, sterilizing and releasing large numbers of males to mate with<br />
wild females, who will then produce unviable eggs. Hence, sterile medfly males can be<br />
considered as beneficial organisms within an integrated pest management program. Sterile<br />
males released may directly be exposed to pesticides used against other citrus pests in the<br />
target area. Therefore, the side-effects of these pesticides should be assessed on these males.<br />
Methods: Laboratory trials were conducted to find out side-effects of the pesticides abamectin,<br />
chlorpyrifos, etofenprox, etoxazole, narrow-range mineral oil, spinosad, pymetrozine and<br />
pyriproxyfen on Vienna-8 tsl sterile C. capitata males. Three-day-old sterile males were<br />
exposed to fresh, 7, 14 and 21-day-old residues of the selected pesticides. Mortality of sterile<br />
males was observed daily during 6 days after pesticide application. When differences against<br />
the control were found, the resulting corrected mortalities (Abbott) were interpreted according<br />
to IOBC standards.<br />
Results: All pesticides resulted harmless to Vienna-8 tsl sterile males except for chlorpyrifos<br />
and spinosad. Both pesticides resulted moderate harmful according to the IOBC categories for<br />
the fresh residue and decreased to slightly harmful until residue of 14-d-old. At 21-d-old<br />
residue, chlorpyrifos resulted harmless whereas spinosad continued as slightly harmful.<br />
Conclusions: Our results provide useful information to implement an SIT program within an<br />
citrus area where an IPM program is being conducted. Sub-lethal effects of pesticides on sterile<br />
Vienna-8 strain of C. capitata males mating behavior should also be considered for a complete<br />
analysis of their impact.<br />
Keywords: Ceratitis capitata, sterile insect technique, insecticide toxicity.<br />
291
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FIELD EVALUATION OF GF-120 TO CONTROL CERATITIS CAPITATA (WIEDEMANN)<br />
IN IRAQ.<br />
Khlawi, Samira & Mohammed Ali, Hussain<br />
Ministry of Science & Technology, Directorate of Agricultural Research, P.O.Box 765 Baghdad, Iraq. E-mail :<br />
halrubeai@yahoo.com.<br />
Background : The medfly, Ceratites capitata has been recorded for the first time infested<br />
orchard citrus fruits in late 2006, and since then spread to all areas of orange grooves and<br />
become one of the major pest in Iraq. This because of citrus fruit importation from neighboring<br />
countries known to have this pest for decades and for inefficient national quarantine system.<br />
The crises depicted the urgent need for the implementation of national control campaign,<br />
taking in mind using effective alternatives to harsh insecticides with low environmental risk.<br />
GF-120 consist of spinosad as an active ingredient, derived from bacteria, and mixed with<br />
attractant and feeding stimulant agents, proven to be effective in controlling many pest species<br />
including medfly with a low mammalian and wildlife toxicity.<br />
Materials & Methods : The efficacy of GF-120 ( Spintor Fly 0.24 g/l, Dow AgroSciences) was<br />
evaluated against C. capitata in citrus orchards of Iraq. Trials were carried out in mandarin<br />
oranges orchard at Baghdad in the year of 2007, and in naval, mandarin and sour orange<br />
orchard at Wasset province ( 50 km east of Baghdad) in the year of 2008. For all treatments<br />
FG-120 was applied as spot treatment in a concentration of 300ml/5 L water. Treatment of<br />
2007, the solution was applied two times with 10 days interval using 15 L handbach spryer.<br />
Twenty five trees (replicates) were treated each time, in addition to 10 trees for the control.<br />
Differences between treatments were determined by checking, at 10 days after treatment and at<br />
harvest time, 100 fruits randomly picked. Treatment of 2008 was applied at three times: before<br />
adult emergence, after adult emergence and after 10 days of the 2 nd treatment. The experiments<br />
were designed to have four groups of trees, each with 25 trees and the control. The 1 st group<br />
treated 3 times, the 2 nd treated twice, the 3 rd group treated once and the control group. At the<br />
end of the season, percentages of infested fruits were determined by checking of 100 fruits for<br />
each group.<br />
Results: The result of the experiments carried out in 2007 on orange mandarin indicated lower<br />
percentage of damaged fruits (5-8%) in treatment with GF-120 compared with untreated check<br />
(21-36%).In the trial of 2008, on mixed oranges treatment resulted statistically different<br />
between treatment timing and number. Percentage of infestation achieved were 3,5,10% for 3,2<br />
and one time treatments ,respectively. All treatments result showed significant difference<br />
compared to the untreated check (89%).<br />
Conclusion: Results achieved in all trails showed that spot foliar surface treatment with GF-<br />
120 is a promising product to control the medfly, C. capitata with low impact on the<br />
environment.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
POPULATION MONITORING OF CERATITIS CAPITATA (DIPTERA: TEPHRITIDAE) IN<br />
S. MIGUEL ISLAND IN THE LAST FIVE YEARS.<br />
Medeiros, Aida 1 ; Tavares, Laura 1 & Oliveira, Luísa 2<br />
1 Direcção Serviços de Agricultura e Pecuária – Direcção Regional Desenvolvimento Agrário – Quinta de S.<br />
Gonçalo, 9500-343 Ponta Delgada, Açores, Aida.MC.Medeiros@azores.gov.pt; 2 Departamento de Biologia, CBA,<br />
CIRN, Universidade dos Açores, Rua da Mãe de Deus, 9501-801 Ponta Delgada, Açores.<br />
Background: To develop a control program against the Mediterranean fruit fly, or medfly,<br />
Ceratitis capitata (Wiedemann), is essential to know the population size and presence around<br />
the year to establish the programs. Population monitoring is a key point in control programs.<br />
The extent of lures and traps available in the market pushed us to establish a pilot program to<br />
select the best trap and lure for the San Miguel Island.<br />
Methods: Monitoring was conducted through 2005-2009 in three fruit orchards in the central<br />
part of São Miguel Island, Azores. One orchard was a monoculture orchard and the other two<br />
were mixed-fruit orchards. Monitoring used “Easy-trap”, with two different attractants: the<br />
male specific pheromone trimedlure and a food-based attractant – “Ferag” lure (ammonium<br />
acetate + diaminoalcane + trimethylamine). Two traps, one with each attractant, were placed in<br />
each orchard. Traps were suspended in the tree canopy.<br />
Results & Conclusions: In the two mixed-fruit orchard adults were captured year-round with a<br />
break during March-May. In the monoculture orchard, however, captures were limited to July-<br />
December. Significant differences were observed between the two types of attractants and<br />
among the years, independently of the location of the orchard. The mean number of adults per<br />
day captured in the pheromone trap was significantly higher than traps baited with Ferag lure.<br />
However, using both attractants increases the potential of controlling the fly infestations by<br />
better predicting the presence and the numbers of the flies.<br />
Keywords: Med Fly, monitoring, attractants<br />
293
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ECONOMIC IMPORTANCE OF THE MEDITERRANEAN FRUITFLY, CERATITIS<br />
CAPITATA IN THE ORCHARDS OF CENTRAL IRAQ.<br />
Mohammed Z. Khalaf ;Adnan H. Salman ;Ali K. Shbar and Farqad A. Sadq<br />
Integrated Pest Control Research center, Ministry of Science & Technology. Baghdad, IRAQ Email:<br />
mzkhalaf2007@yahoo.com<br />
Background: In the recent years, The Mediterranean Fruitfly Ceratitis capitata is distributed in<br />
the orchards of central Iraq, and caused highly economic losses.<br />
Methods: The study was conducted in orchards in central Iraq during 2009 and make field<br />
survey of the insect in four types of orchards (citrus, Apricot, Figs & Citrus, a mixture of fruit<br />
trees), and used for this purpose Tefri traps supplied with Q-lure and DDVP.<br />
Results: The results of the study for the presence of the insect throughout the year ,and reached<br />
its highest numerical density of the insect in citrus orchards during the month of October and<br />
November were ( 295 – 870 ) insect per trap per 2 week and the least numerical density during<br />
the months of January and February .While the numerical density of the insect in the orchards<br />
of Apricot ,Figs & Citrus ,a mixture of fruit trees were 9 ,166 , 207 Insect per trap per 2 week<br />
each respectively, during the months of October and November.<br />
Conclusions: The C. capitata caused highly economic losses in citrus orchards reached 60% of<br />
the mandarin fruits .Currently in Iraq to fight no control method to reduce the economic losses<br />
caused by this pest except the use of pesticides GF-120.<br />
Keywords: Fruitfly , Survey , Iraq
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
ELECTROANTENNOGRAPHY FOR EVALUATION OF FOOD-BASED ATTRACTANTS<br />
FOR PEST TEPHRITIDAE.<br />
Kendra, Paul E.; Montgomery, Wayne S.; Epsky, Nancy D. & Heath, Robert R.*<br />
United States Department of Agriculture, Agricultural Research Service, Subtropical Horticulture Research<br />
Station, 13601 Old Cutler Road, Miami, Florida, 33158, USA. Email: paul.kendra@ars.usda.gov<br />
Background: Anastrepha fruit flies are serious economic pests of fruit crops throughout the<br />
American tropics and subtropics. Current trapping systems utilize synthetic lures that emit<br />
ammonia and other attractant chemicals that function as protein feeding cues. However, fly<br />
captures are variable with synthetic lures, and this variability does not appear to be correlated<br />
solely with ammonia release rate. The need for improved attractants prompted research using<br />
electroantennography (EAG), a technique that measures response of antennal olfactory<br />
receptors to volatile chemical stimuli.<br />
Methods: All EAG analyses were conducted with a Syntech system (Hilversum, The<br />
Netherlands) using laboratory-reared adults (of known age and physiological state) of the<br />
Caribbean fruit fly, Anastrepha suspensa (Loew). Antennae were presented with quantified<br />
chemical samples in saturated vapor form, and EAG responses were recorded and normalized<br />
relative to a standard reference chemical (20 µl 2-butanone).<br />
Results: To date, our group has quantified antennal response of male and female A. suspensa to<br />
ammonia, carbon dioxide, ammonium bicarbonate, 2-butanone, 3-methyl-1-butanol,<br />
putrescine, cadaverine, and homologous terminal diamines. Analysis of the amplitude of EAG<br />
response has provided information on appropriate doses and combinations of chemicals needed<br />
to elicit optimal antennal response. Comparative EAG has identified several factors that<br />
contribute to the variability in fly response to known food-based attractants, including sex, age,<br />
nutritional requirements, and reproductive status. In addition, EAG has provided insight into<br />
the antennal chemoreceptors involved in detection of semiochemicals, and has identified<br />
potential new fruit fly attractants.<br />
Conclusions: Quantitative electroantennography is a valuable tool for evaluation of peripheral<br />
olfactory response of tephritid flies to potential attractants. Information obtained from EAG<br />
analysis will be used to determine relationships between antennal sensitivity to food-based<br />
attractants and efficacy of those compounds when deployed in traps for capture of pest species.<br />
Keywords: Anastrepha suspensa, electroantennography, olfaction, synthetic lures, food-based attractants<br />
295
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
DEVELOPMENT OF SPLAT MAT CONTROLLED RELEASE SEMIOCHEMICAL BAIT<br />
FORMULATIONS FOR LONG LASTING FRUIT FLY MANAGEMENT<br />
Mafra-Neto, Agenor* 1 ; Gomez, Luis E. 2 ; Vargas, Roger 3 ; Stoltman, Lyndsie 1 ; Zeni,<br />
Diego 1 , Borges, Rafael 1<br />
1 ISCA Technologies, 1230 Spring St., Riverside California 92507, US. Email: president@iscatech.com; 2 Dow<br />
AgroSciences LLC Indianapolis, Indiana US. 3 USDA, ARS, Hilo, Hawaii US.<br />
Background: The insecticide landscape is changing rapidly in orchard crop production in the<br />
US and abroad. This is particularly true for nursery, greenhouse, fruit, olive and nut production<br />
where conventional insecticide products that form the foundation of current management<br />
programs are being lost. Small growers in particular, suffer with this increasing lack of<br />
available control tools and from pressure of suburban neighbors to reduce use of conventional<br />
insecticides. Current Male Annihilation Techniques (MAT) combine male-specific attractants<br />
with insecticide in traps and devices that, while effective, require routine service that is costly<br />
and labor intensive. ISCA Technologies’ Specialized Pheromone and Lure Application<br />
Technology (SPLAT) was initially developed for the mechanical deployment of small doses of<br />
Lepidopteran pheromones for long lasting mating disruption. Here we report further<br />
development of this biologically inert matrix to create Tephritid management formulations that<br />
are hand or mechanically applied, rain-fast, and provide long-term controlled release of a<br />
variety of semiochemicals and pesticides at biologically relevant levels.<br />
Methods: SPLAT was formulated with spinosad, malathion, naled and other insecticides along<br />
with attractants such as methyl eugenol, trimedlure, cue-lure, raspberry ketones, and other<br />
parapheromones, attractants and feeding stimulants to achieve specific killing action against a<br />
variety of Tephritidae, in both laboratory and field trials. Studies have been conducted to<br />
determine the longevity and mortality of SPLAT formulations for fruit flies resulting from<br />
variable point source densities, sizes and concentrations of active ingredients. Methods of<br />
SPLAT application ranged from hand-held caulking tubes and electric grease guns to<br />
mechanical and aerial applications.<br />
Results: Field trials conducted in Hawaii showed that SPLAT could be combined with naled<br />
and spinosad, resulting in significantly higher mortality of B. dorsalis males with greater<br />
longevity than other baits and thickener formulations. Field trials in tropical urban areas<br />
indicate that a single application of SPLAT combined with methyl eugenol and Spinosad<br />
effectively controls B. carambolae for months. Studies with A. ludens, C. capitata and A.<br />
fraterculus have shown that SPLAT combined with spinosad, feeding stimulants and<br />
attractants resulted in up to 99% mortality within hours. Laboratory data also show that SPLAT<br />
is amenable to the inclusion of a wide range of active ingredients, including volatile alcohols,<br />
acetates and aldehydes which can increase attraction for some fruit fly species. SPLAT matrix<br />
provided protection of active ingredients tested and increased longevity, a particularly<br />
important issue for areas with high temperatures and rainfall.<br />
Conclusions: Due to its amorphous and flowable quality, SPLAT MAT provides a mechanized<br />
attract and kill formulation for long-term farm-wide as well as area-wide management and<br />
control of populations of a variety of Tephritid fruit fly species. The size and density of point<br />
sources can be tailored to treat any sized field or pest density.<br />
Keywords: SPLAT, Spinosad, Attract-and-kill, Attractant, Mortality
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
MEDITERRANEAN FRUIT FLY, CERATITIS CAPITATA (WIEDEMANN) MASS-<br />
TRAPPING ON CLEMENTINE GROVES IN SPAIN: OPTIMIZING TRAP DENSITY.<br />
Martínez-Ferrer, María Teresa*; Campos, José Miguel & Fibla, José M.<br />
IRTA Amposta. Carretera de Balada, Km 1. E- 43870-Amposta (Tarragona), Spain. Email:<br />
teresa.martinez@irta.es.<br />
Background: In citrus, C. capitata mainly causes damage to early ripening varieties, such as<br />
Clementine (Citrus reticulata Blanco). Up to 8 sprays can be performed in a grove to control<br />
Medfly in a season. The mass trapping strategy attempts to provide a successful pest control.<br />
The system is based on placing a high amount of traps with an attractant and a toxicant, aiming<br />
to capture as large as possible number of adults in the field. In Spain, more than 30,000 ha of<br />
citrus groves protected against Medfly by mass trapping showed good efficacy using a density<br />
of 50 traps per ha during 3 months before harvesting. Our aim is to study the efficacy of this<br />
method specifically on the most abundant Clementine variety and optimize the trap density<br />
required to successfully control this pest.<br />
Methods: Six commercial groves of Clemenules (Citrus reticulata Blanco cv. Clemenules)<br />
citrus variety of approximately 3 ha in size were divided each one into three plots and each plot<br />
was assigned a different trap density: 25 (D25), 50 (D50) and 75 (D75) traps per ha.<br />
Probodelt ® (Maxitrap ® model) traps with the attractant Ferag CC D TM ® (SEDQ), comprising<br />
three membrane dispensers of Trimethylamine, Ammonium acetate and Diaminoalkane, were<br />
placed in position. DDVP was used as toxicant. Colour rind index, internal maturity of fruits,<br />
adults captured in traps, and fruit damage were assessed.<br />
Results: Fruits from all groves and years were ready to harvest at the end of October, according<br />
to commercial quality parameters required for trade export. The higher the trap density, the<br />
lower the number of adult captures per trap and day with, on average, 1.21 ± 0.13, 0.27 ± 0.02<br />
and 0.23 ± 0.02 for D25, D50 and D75, respectively. However, the number of adults captured<br />
per ha was similar for D50 and D75 trap densities, while both were significantly lower than<br />
D25. The average number of adults captured was 270.2 ± 69.9, 123.3 ± 28.7, and 152.3 ± 53.7<br />
for D25, D50 and D75, respectively. For all the years and plots, the maximum percentage of<br />
fruits attacked was 1.5%. The number of attacked fruits was similar between plots. Damaged<br />
fruits on the perimeter rows were significantly higher than inside the plots.<br />
Conclusions: Mass trapping is a valid stand-alone control method against C. capitata in<br />
varieties of Clementine that start maturing by October, and a 25 traps per ha density is<br />
sufficient to ensure successful control of this pest. Therefore, our findings imply that a mass<br />
trapping control program, using 25 traps per ha, can be implemented providing protection<br />
against med fly over 50% of production in our citrus producing area.<br />
Keywords: mass trapping, trap density, fruit fly, citrus.<br />
297
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
FRUIT FLY MANAGEMENT IN OLIVES WITH A NOVEL INNOVATIVE BIOLOGICAL<br />
SYSTEM (GF-120 ® ).<br />
Mavrotas, Costas* 1 ; Paroonagian, Doris 2 ; Gomez, Luis E. 2 ; Torné, Maria 3 . Tescari,<br />
Enzo 4 , Lysandrou, Michael 1<br />
1 Dow AgroSciences, Thorikon – Lavrion, 19500, Attika, Greece, cmavrotas@dow.com; 2 Dow AgroSciences LLC,<br />
Indianapolis, IN, USA., 3 Dow AgroSciences, Madrid, Spain, Dow AgroSciences, Bologna, Italy.<br />
Background: The olive fruit fly Bactrocera (=Dacus) oleae, is the most destructive pest of<br />
olives. Crop losses, quantitative and qualitative, due to this pest can reach up to 100% for table<br />
cultivars and 80% of oil value if no control measures are taken. In the last forty years a variety<br />
of methods have been used to control this pest and protect against significant economic yield<br />
losses. Current control methods include full cover, protein-based bait sprays and SIT. Bait<br />
applications have proven to be much safer to the crop and environment than the full cover<br />
sprays. GF-120 ® (SPINTOR ® CEBO, SUCCESS ® , SPINTOR ® FLY) is a concentrated premixed<br />
fruit fly bait containing a reduced risk toxicant to both mammals and non-target insects.<br />
Attractants in GF-120 have been optimized to control a plethora of the most damaging and<br />
invasive fruit fly species. GF-120 ® attracts and controls male and female tephritid fruit flies. It<br />
contains an optimized blend of feeding attractants and a green chemistry insecticide, spinosad;<br />
which is derived from a naturally occurring soil bacterium, Saccharopolyspora spinosa.<br />
Methods: Field and laboratory studies have been conducted in olives, throughout the main olive<br />
producing countries aiming to evaluate the efficacy against olive fruit fly and the medfly,<br />
selectivity, attractiveness, residuality, potential risk of resistance and impact on non-target<br />
organisms of GF-120 ® .<br />
Results: GF-120 ® applied at the rate of 1.0 -.1.6 L pr/ha as a true bait showed that it is very<br />
effective against olive fruit fly and the medfly compared to the traditional control programs<br />
consisting of tank mixes of organophosphate or pyrethroid with hydrolyzed protein. Optimized<br />
bait application technology targets achieving large droplet size (4-6 mm) which results in<br />
improved attractiveness and residuality. The potential risk of resistance development was<br />
evaluated in six consecutive generations and proved that it is very low. GF-120 ® impact on<br />
non-target organisms (NTOs) has been evaluated in a number of studies confirming greater<br />
levels of selectivity compared to traditional fruit fly control solutions (organophosphate or<br />
pyrethroid with hydrolyzed protein). GF-120 ® is approved for use in organic farming and has<br />
an excellent environmental profile and has been demonstrated under laboratory, and field<br />
conditions to be an ideal product for use in IPM programs and sterile insect technique (SIT)<br />
control strategies.<br />
Conclusions : Years of extensive research trials and commercial use across the globe have<br />
proven that GF-120 ® applied either by ground or by air at 1.0-1.6 lt pr/ha targeting true bait<br />
applications provides the most effective, safe and organic solution for the olive industry.<br />
Keywords: Spinosad, GF-120 ® , Bactrocera oleae, Olive fruit fly, insecticidal baits.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
COMPARATIVE STUDIES OF THE EFFECTIVENESS AND ATTRACTIVENESS OF THE<br />
TRAP "SENTRAP"AND TWO HOME-MADE PROTOTYPES TO "MULTULURE"<br />
(CONVENTIONAL TRAP) IN MALE ANIHILATION TECHNIQUE.<br />
Mbaye, Ndiaye 1 & Souleymane, Lessueur 2<br />
1 National Plant Protection Service ; Km 15, Route de Rufisque. Box: 20054 Thiaroye, Dakar. Republic of<br />
SENEGAL. Email : E-mail : mbaye1314@hotmail.fr; 2 SENCHIM section recherches et développement Dakar<br />
Bacground: If pests could be attracted to few points where they would be either in contact with<br />
or consume the toxin then many of the objections that confront broadcasted pesticides could be<br />
overcome. Studies have shown that peak population of Bactrocera invadens, the most<br />
destructive invading fruit fly species, occurs during the wettest periods with an abundance of<br />
food. So, such situation limits the use of killer-blocks in open air in Male Annihilation<br />
technique (MAT). This study aims to compare the effectiveness and attractiveness of these<br />
different local trapping devices to the conventional "Multilure" (Better World Manufacturing,<br />
Inc.., Miami, FL) regarding partly the limited financial resources of producers.<br />
Methods: The study was conducted in a 10-years-old mango orchard of fifty six (56) ha of the<br />
late season variety "Kent"in Senegal. It was set in 3 randomized blocs and treatment repeated 4<br />
times with 50m between traps. Killer-blocks (5 x 5 x 1,25 cm) of Triplex are soaked in<br />
Mal'atrap (75% methyl eugenol and 25% malathion). Flies were collected separately every 10<br />
days. The experiment lasted from July 15 to October 15, 2008. Data variability on individuals<br />
were analyzed using a one-way ANOVA, means separation between treatments performed by<br />
the Tukey honestly significant difference (HSD) test at 5%.The dependence with variables<br />
such as time and catch was done using the test of Pearson correlation.<br />
Results: Average catches of "Multilure" to those of 1.5-liter, "Sentrap" and 5-liters showed a<br />
significant difference (F = 5.472, df 3, α = 0.003). However, Results from the Tukey HSD test<br />
indicated that only 5-liters trap was statistically comparable to "Multilure". The study of the<br />
relationship between catch at different times and type of trap showed strong relationships.<br />
Sentrap, and "Multilure" exhibited a very significant dependency relationships to the variables<br />
time and catch (P=-0.969** and P=-0.903**; α < 0.01); the common feature between these two<br />
traps is their yellow basal color.<br />
Conclusions: In case of absence of conventional traps, the plastic bottle of 5 liters could,<br />
therefore, be used as an alternative and its basal part painted in yellow should increase its<br />
efficiency. Due to simple design and less time needed for servicing "Sentrap", increasing its<br />
volume and openings and fixing firmly the upper part with a lid screw might improve its<br />
efficiency.<br />
Keywords: Traps, catches, Bactrocera<br />
299
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
PRELIMINARY RESULTS OF AN INTEGRATION PEST MANAGEMENT PROGRAM OF<br />
FRUIT FLIES (DIPTERA: TEPHRITIDAE) ATTACKING MANGOES IN CENTRAL<br />
TANZANIA.<br />
Mwatawala, Maulid Walad 1* ; Maerere, Amon Petro 1 ; De Meyer, Marc 2 & Rwegasira,<br />
Gration 1<br />
1 Sokoine University of Agriculture (SUA), Department of Crop Science and Production, Box 3005, Chuo Kikuu,<br />
Morogoro, Tanzania. mwatawala@yahoo.com. 2 Royal Museum for Central Africa (RMCA), Entomology section,<br />
Tervuren, Belgium.<br />
Background: There is no proper management program for fruit flies attacking mangoes, despite<br />
heavy losses they inflict on the crop, especially after the introduction of Bactrocera invadens.<br />
There is a need for testing a number of management programs on their efficacies in reducing<br />
losses due to fruit flies.<br />
Methods: Trials were conducted in eight selected orchards located in the mango growing areas<br />
of Morogoro Region, Central Tanzania. The treatments included (i) orchard sanitation (ii)<br />
orchard sanitation, spray of dimethoate 480EC and early harvesting (iii) orchard sanitation,<br />
spray of GF 120 containing 0.02% spinosad, male annihilation using methyl eugenol and early<br />
harvesting (iv) spray of a locally formulated bait (containing molasses, browsers’ waste and<br />
crude extracts of Derris elliptica as a toxicant) and early harvesting. Each treatment was<br />
applied in an individual orchard and replicate twice. Additionally, two orchards of unimproved<br />
mango varieties and not subjected to any fruit flies management method were included in the<br />
study. Populations of fruit flies in each orchard were monitored using torula yeast placed in<br />
McPhail traps. The replication of traps within each site were sub-sampled to construct point<br />
wise biased and accelerated bootstrap confidence intervals that were used to compare<br />
treatments. Further, fruits were sampled at ripening and individually placed in containers to<br />
determine incidences and infestation rates of emerged fruit fly species. Analysis of Variance<br />
(ANOVA) was used to compare infestation rates while chi-square was used to compare<br />
incidences of fruit flies. Log-linear regression models were used to correlate population of<br />
fruit flies, season and weather parameters recorded at each orchard.<br />
Results: Different management programs have shown differences in their efficacies in reducing<br />
losses due to fruit flies.<br />
Conclusion: Losses due to fruit flies can be reduced using ecologically based management<br />
programs. However, further refinement of some programs is still needed.<br />
Keywords: IPM, fruit flies, mangoes, Tanzania
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
ADRESS® COUPLED WITH SIT: AN INTEGRATED STRATEGY TO CONTROL<br />
MEDITERRANEAN FRUIT FLY.<br />
Navarro-Llopis, Vicente*; Sanchis, Juan; Vacas, Sandra, Alfaro Cañamás, Cristina;<br />
Primo, Jaime.<br />
Centro de Ecología Química Agrícola-Instituto Agroforestal del Mediterráneo, Universidad Politécnica de<br />
Valencia, Edificio 6C, Camino de Vera s/n 46022, Valencia, Spain. Email: vinallo@ceqa.upv.es<br />
Background: Sterile Insect Technique (SIT) has been implemented over Wide Areas all around<br />
the world. SIT best results were obtained when Ceratitis capitata population was low or very<br />
low and this is key point to ensure success of this technique. For this purpose combination of<br />
SIT with natural enemies, insecticide sprays or lure and kill technique has been studied in last<br />
decades. In this way, we propose to combine chemosterilization technique using Adress®<br />
devices with SIT as both affect medfly population reducing fertility. In this case,<br />
chemosterilization affects directly wild population and does not kill sterile released individuals<br />
and this can suppose a great advantage. Results of combination of both techniques over a wide<br />
area compared with only SIT treated area are showed.<br />
Methods: During 2008 and 2009, the efficacy of the combination of two fruit fly control<br />
techniques, SIT and Adress®, was tested in three crops: citrus, stone fruit and persimmon.<br />
2000 sterile males were released per ha and week in the whole trial area (50.000 ha, SIT area).<br />
Twenty four Adress® devices per ha were hung in 3600 ha (SIT+Adress® area) inside of the<br />
whole trial area,. Ten SIT+Adress® plots per each crop and 10 SIT plots per each crop were<br />
assessed throughout the trial period. In order to evaluate the efficacy, the male and female<br />
population was monitored from August 2008 to November 2009 and injured and the viability<br />
of the egg-laying was assessed.<br />
Results: Results showed an important reduction in the C. capitata population in plots treated<br />
with both techniques versus plots treated only with SIT. A reduction in the percentage of<br />
injured fruit in SIT+Adress® plots was observed and the viability of the egg-laying was also<br />
minor.<br />
Conclusions: Results show the compatibility of these techniques and provide the possibility of<br />
use Adress® coupled SIT to reduce C. capitata population in locations with high fruit fly<br />
population.<br />
Keywords: Ceratitis capitata, chemosterilization, Adress, SIT, sterile<br />
301
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
OLFACTORY RESPONSES OF MALE MEDFLIES TO PLANT MATERIAL CONTAINING<br />
THE PARAPHEROMONE Α-COPAENE.<br />
Niogret, Jerome; Epsky, Nancy D.*; Montgomery, Wayne S.; Kendra, Paul E. & Heath,<br />
Robert R.<br />
United States Department of Agriculture, Agricultural Research Service, Subtropical Horticulture Research<br />
Station, 13601 Old Cutler Road, Miami, FL 33158-1857 USA. Email: Jerome.niogret@ars.usda.gov<br />
Background: The Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera:<br />
Tephritidae) is a highly invasive species that is considered the most adaptable and polyphagous<br />
species of tephritid fruit fly due to its global distribution and its broad range of host plants,<br />
primarily tropical and subtropical fruits and vegetables. Ceratitis capitata is presently ranked<br />
first among economically important fruit fly pests, due to both damage to crops and costs of<br />
eradication. Trimedlure is a synthetic chemical that is highly attractive to male medflies and is<br />
the standard male-targeted lure used for this species. Medfly response to trimedlure is similar<br />
to response to the sesquiterpene α-copaene, a widely-distributed plant compound, and males<br />
respond to both host and non-host sources that contain α-copaene. Although α-copaene is<br />
reported to be 2 to 5 times more attractive than trimedlure, difficulties in obtaining synthetic αcopaene<br />
in sufficient quantities for large-scale trap deployment have prevented its use as a lure.<br />
As part of a study on sesquiterpene content of tree cambial tissue, we found that cambial<br />
(cambium+bark layers) tissue from avocado, Persea americana, contained α-copaene, and that<br />
levels were highly variable among different genotypes. Therefore, studies were initiated to<br />
determine if these sources of α-copaene were biologically active for male medflies. For<br />
comparative purposes, results were compared with responses to cambial tissue from Litchi<br />
chinensis and Ficus benjamina, two substrates known to elicit behavioral responses in male<br />
medflies.<br />
Methods: Behavioral bioassays and electroantennography (EAG) were used to evaluate<br />
responses of sterile male medflies; and GC-MS analysis was used to quantify the amounts of<br />
13 sesquiterpenes, including α-copaene, common in cambial tissue from four avocado<br />
genotypes, from Litchi and from Ficus.<br />
Results: Litchi elicited the highest response and Ficus the lowest response, with cambial tissue<br />
from the avocado genotypes eliciting intermediate responses that varied significantly among<br />
the four types in both the bioassays and EAG experiments. These responses, however, were not<br />
correlated with the amount of α-copaene, but were correlated with few other components like<br />
a-humulene, which triggered strong antennal responses in EAG.<br />
Discussion: Additional sesquiterpenes may be responsible for the high responses observed with<br />
the low α-copaene substrates. Identification of these chemicals may provide a new<br />
understanding of the biological basis for the response of male medflies to these wood sources,<br />
which could lead to development of new tools for improved detection and control.<br />
Attractiveness of α-humulene still has to be tested in bioassays but, this compound triggered a<br />
strong antennal response of the male medflies. Although not necessarily attractive, this<br />
compound may play a synergetic role to α-copaene for C. capitata attraction.<br />
Keywords: Ceratitis capitata, bioassays, Electroantennography, olfaction, cambial
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
USING GIS TECHNOLOGY IN CONJUNCTION WITH FRUIT FLY TRAPPING SURVEY<br />
IN THAILAND.<br />
Noikeaing, Sirilux *1 , Piyarath, Sudsa-nguan 1 ; Sittipitak, Wanlapa 1 & Orankanok,<br />
Watchreeporn 2<br />
1 Department of Agriculture Extension, Ministry of Agriculture and Cooperatives; 2143/1 Phahol Yotin Road,<br />
Chatuchak, Bangkok, 10900, Thailand; Email: siriluxr@gmail.com<br />
Background: Geographic Information System, GIS, has been used to support area-wide fruit<br />
fly control base on the Sterile Insect Technique, SIT, and related environment-friendly<br />
technology in Thailand for 10 years. As a tool, GIS has been used to define project areas, core<br />
area and buffer zone, mark trapping site for monitoring fruit fly population dynamic.<br />
Differrence distance of trapping site were discussed in vision of GIS tehnician and operation<br />
cost.<br />
Methods: Two tephritid species (Diptera: Tephritidae), the Oriental fruit fly Bactrocera<br />
dorsalis (Hendel) and the guava fruity fly Bactrocera correcta (Bezzi) have been considered to<br />
be the key insect pests which covered over 160 km 2 of two areas in Phichit province, one area<br />
in Ratchaburi province and one area in Prachuap Khiri Khan province. Steiner traps with<br />
methyl eugenol as fruit fly attractant were design as 1 trap per 1, 2, 4, 8 and 10 square<br />
kilometers. Traps located were marked by GIS program therefore relalted distance of radius<br />
were 1, 1.414, 2, 2.83 and 3.16 km, respectively. Trapping network weekly deployment<br />
frequency was conducted 52 weeks continueously. Fruit fly population idex was showed in<br />
term of fly/trap/day, FTD.<br />
Results: In four study areas, GIS program genarated 158 trap points from trap density in one<br />
square kilometers, 80 trap points from trap density in two square kilometers, 36 trap points<br />
from trap density in four square kilometers, 18 trap points from trap density in eight square<br />
kilometers and 15 trap points from trap density in 10 square kilometers. The result showed that<br />
the trap points from trap density in one square kilometers and two square kilometers covered a<br />
whole project area but the others did not cover a whole project area. Ranking FTD in seven<br />
was classed using ISPM standard in the first three classes and the others classes were classified<br />
by the calculation of total data and the expert.<br />
Conclusions: Area-wide Sterile Insect Technique (SIT) programs in a cost-trapping can be a<br />
trapping radius of approximately 1.414 kilometers and/or the trap density in two square<br />
kilometers.<br />
Keywords: GIS, SIT, fruit fly control, trapping survey, Thailand.<br />
303
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF TRAP DENSITY ON NUMBER OF CAPTURED FRUIT FLIES.<br />
Orankanok, Somchai* 1 ; Amornsak, Weerawan 2 & Orankanok, Watchreeporn 3<br />
1 Department of Animal Sciences, Faculty of Agriculture and Natural Resources, Rajamangala University of<br />
Technology Tawan-Ok (RMUTTO), Chonburi. 20110. Thailand. . Email: orankanoks@hotmail.com; 2 Department<br />
of Entomology, Faculty of Agriculture, Kasetsart University, Bangkhen Campus, Chatuchak, Bangkok, 10900,<br />
Thailand; 3 Department of Agriculture Extension, Ministry of Agriculture and Cooperatives, 2143/1 Phahol Yotin<br />
Road, Chatuchak, Bangkok, 10900, Thailand<br />
Background: Fruit fly control has been listed in International Standard of Phytosanitary<br />
Measure, ISPM. Surveillance process, data interpretation and assessment are major important<br />
systems required in order to thoroughly understand nature of pests and use information as a<br />
tool for making decision in conducting activities. Currently, fruit fly control program in<br />
Thailand needs much more attention in various aspects than traditional control-based<br />
knowledge. Fruit fly investigation is mainly to reduce expense of trap surveillance latest<br />
technologies, sterile insect technique, biological control, bait application technique and male<br />
annihilation.<br />
Methods: The modified steiner traps using methyl eugenol as an attractant were placed as following:<br />
a trap/km 2 , a trap/2 km 2 , a trap/4 km 2 , a trap/8 km 2 and a trap/10 km 2 , in four fruit orchard areas, Pak-<br />
Tau district in Ratchaburi province, Pohpratunchang district in Pichit province, Saklek district in<br />
Pichit province and Pranburi district in Prachuabkirikhan province. The total area was 160 km 2 with<br />
307 traps. Fruit flies were continuously trapped for each seven days/week of 52 weeks. Collected<br />
trap data were 31,928. The index to indicate fruit fly population is number of fruit fly/trap/day.<br />
Results: The interaction of the treatments based on fruit fly number and trap density in each area<br />
depended on geographical location and fruit fly species. Distribution of B. dorsalis at Pranburi<br />
district in Prachuabkirikhan province was shown higher than other areas significantly (p
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EFFICACY OF KAOLIN AND COPPER TO CONTROL BACTROCERA OLEAE (ROSSI)<br />
AND SIDE EFFECTS ON NON-TARGET ARTHROPODS.<br />
Pascual, Susana 1* ; Cobo, Ana 1 ; Marcotegui, Aránzazu 1 ; Sánchez-Ramos, Ismael 1 ; Seris,<br />
Elena 1 ; Cobos, Guillermo 1 ; Bengochea, Paloma 2 ; González-Núñez, Manuel 1<br />
1 Departamento de Protección Vegetal, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria<br />
(INIA). Carretera de A Coruña km 7,5 28040 Madrid, Spain. Email mgnunez@inia.es. 2 Departamento de<br />
Producción Vegetal: Botánica y Protección Vegetal. Escuela Técnica Superior de Ingenieros Agrónomos<br />
(ETSIA), Universidad Politécnica de Madrid, Madrid, Spain.<br />
Background: Olive fruit fly, Bactrocera oleae, is the key pest of the olive crop, demanding<br />
annual treatments. Therefore, the development of environmentally friendly control methods is<br />
of growing interest. Results of the evaluation in field assays about the efficacy of preventive<br />
application of kaolin and copper to control B. oleae are hereby reported, as well as the side<br />
effects of both treatments on non-target arthropods, paying special attention to natural enemies.<br />
Methods: The field trial was conducted in 2008 in an olive grove at Villarejo de Salvanés,<br />
Madrid, using a randomized complete block design with four replicates. Treatments consisted<br />
of three applications of kaolin (Surround � WP), copper (Bordeaux mixture, Cuprocal � WP)<br />
and dimethoate bait sprays (Danadim Progres � plus protein hydrolisate, Life � ) as a positive<br />
control. B. oleae damage was assessed by the number of olive fruits attacked and the<br />
developmental stages found. Arthropod fauna of olive tree canopies was sampled by a beating<br />
method throughout the growing season. The effect of treatments on abundance and biodiversity<br />
(number of species and Shannon index) of non-target arthropods was evaluated. Data were<br />
analyzed by means of ANOVA and PRC curves were built to estimate the effect of treatments<br />
on natural enemies.<br />
Results: Population levels of B. oleae were very high and none of the treatments significantly<br />
reduced the percentage of olives attacked. However, a significantly lower number of olive<br />
fruits with advanced developmental stages (L3 or pupae) was registered in kaolin treated olive<br />
trees at harvest time (20% compared to 64% in the control), what resulted in an important<br />
reduction of the damage. No significant changes in abundance and number of species of<br />
arthropods were detected. However, the Shannon diversity index was reduced in dimethoate<br />
bait plots on the two last sampling dates. PRC analysis showed no significant effect of the three<br />
treatments on the natural enemies’ community as a whole but a reduction of some specific taxa<br />
was revealed. Thus, Phylodromidae, Encyrtidae and Coniopterygidae were negatively affected<br />
by the three treatments; kaolin and dimethoate reduced the numbers of the spiders Tmarus spp<br />
and Gibbaranea spp while the populations of the coccinellid beetle Scymnus mediterraneus<br />
were reduced by Kaolin.<br />
Conclusions: Kaolin seems to be able to reduce the damage of B. oleae. However, copper did<br />
not control the fly. We detected some slight deleterious effects on non-target arthropods,<br />
especially in the case of dimethoate. Longer term trials are needed to better characterize both<br />
the efficacy and the side effects of the tested products.<br />
Keywords: Olive fruit fly, kaolin, copper, dimethoate, side-effects<br />
305
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EVALUATION OF MASS TRAPPING EQUIPMENT AGAINST CERATITIS SPP. ON THE<br />
ISLAND OF LA REUNION.<br />
Peñarrubia-María, Esther *1 ; Quilici, Serge 2 ; Schmitt, Chloé 2 ; Vilajeliu, Marià 1 ; Batllori,<br />
Lluís 3 ; Avilla, Jesús 4 & Escudero-Colomar, Lucía Adriana 1<br />
1 Institut de Recerca i Tecnologia Agroalimentàries (IRTA) - Estació Experimental Agrícola Mas Badia. Canet de<br />
la Tallada, 17134 La Tallada d’Empordà, Girona, Spain. esther.penarrubia@irta.cat. 2 Centre de Coopération<br />
Internationale en Recherche Agronomique pour le Développement (CIRAD) Réunion, UMR PVBMT, Saint-<br />
Pierre, France. 3 Servei de Sanitat Vegetal. DAR. Castelló d’Empúries, Girona, Spain. 4 University of Lleida -<br />
Centre UdL – IRTA, Lleida, Spain.<br />
Background: Based on several field trials of equipment used for mass trapping of Ceratitis<br />
capitata conducted in Girona (Spain), similar studies were performed in the island of La<br />
Réunion, where two main Ceratitis spp. (C. rosa and C. capitata) coexist. The aim of the study<br />
was to evaluate in the field the effectiveness of trapping equipment for Ceratitis spp. through<br />
comparative studies of trap types, attractants and insecticides.<br />
Methods: All trials were conducted in commercial Citrus orchards and the experimental design<br />
used randomized blocks with 4 replicates with a monitoring period lasting from 55 to 60 days.<br />
Traps were hung at a height of 1.5 m on Citrus trees. Two rotations per week were made in<br />
order to diminish the possible effect of trap location, giving a total of 5 complete rotations. In<br />
trials on traps and attractants, the number of males and females were recorded while, in trials<br />
on insecticides, alive and dead individuals were taken into account. Data were pooled for each<br />
rotation cycle, transformed (arcsine of the square root of the percentages of captures) and<br />
analyzed through one-way ANOVA (Enterprise Guide, SAS).<br />
Results: Over all the trials, C. rosa was the most abundant Ceratitis species captured.<br />
Maxitrap® and Tephri-trap traps caught similar numbers of C. rosa throughout the trial period<br />
whereas Easy-trap® captured significantly fewer adults. Similar results were obtained for C.<br />
capitata. In the trial on attractants, significantly higher captures of C. rosa were found with<br />
BioLure Med Fly® (3 dispensers) over the study period, followed by Biolure Unipack® and<br />
FERAG® CC D TM. Owing to the lower adults of C. capitata recorded (195) no differences<br />
were found between the attractants tested. In the insecticide test, there were no significant<br />
differences in the numbers of dead C. rosa with DDVP and Deltamethrin, which were higher<br />
than with Alpha-cipermetrine. A significantly lower number of alive adult C. rosa was<br />
observed with DDVP, followed by Deltamethrin. Although captures of C. capitata were lower,<br />
similar numbers of dead flies were observed for all treatments.<br />
Conclusions: The most effective equipment for the capture of C. rosa, the species caught in<br />
higher percentages in all trials, were Maxitrap and Tephri-trap devices, the attractant BioLure<br />
Med Fly and the insecticide Deltamethrin, that could constitute a suitable substitute for DDVP,<br />
recently banned in the EU.<br />
Keywords: Ceratitis capitata, Ceratitis rosa, mass trapping equipment
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
BAIT STATION VERSUS CHEMICAL CONTROL IN ARGENTINA, PERIOD 2007-2008.<br />
Putruele, Graciela 1 & Segade, Gonzalo 2<br />
1 Centro Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto Nacional de Tecnología<br />
Agropecuaria (INTA)-CC 77 (1708) Provincia Buenos Aires. 2 Estación Experimental Agropecuaria San Pedro-<br />
Instituto Nacional de Tecnología Agropecuaria (INTA)-CC 43 (2930) Buenos Aires, ARGENTINA.<br />
Background: In this work, we report the results of comparative field studies on chemical<br />
control, deployment strategy and efficacy of the bait station methods in the control of the<br />
medfly.<br />
Methods: The tests were conducted from November 2007 to June 2008 in San Pedro<br />
(Argentina) with the objective of adding new tools in the integrated pest management (IPM) of<br />
fruit flies. San Pedro is in the North - East of Buenos Aires Province at 33º 39' latitude South<br />
and 59º 41' longitude West at 26.99 m over sea level. Test 1: It was conducted in an orchard of<br />
20 years old (sweet orange var. Valencia late) with density 625 plants/plot (4x4 m). Test 2: In a<br />
6 ha ten-year-old organic orchard (sweet orange var. Washington Navel), with trees spaced<br />
(3x4 m) separately (density 825 plants/plot). On both of them, the following four treatments<br />
were tested (each in a one ha plot within the orchard): a) Mass trapping: One BBS (Gum stick<br />
(classic) 9% AA, 5% TMA, 1% Spinosad, supplied by R. Heath, USDA/ARS, Miami, FL))<br />
placed in every second tree; b) Bait stations: 100 BBS distributed in four rows (25 per row) of<br />
the one ha plot; c) Ground bait sprays (chemical control locally in use) using Spinosad<br />
formulated bait (Flipper) containing 0.02% Spinosad; and d) Untreated control (no control).<br />
Medfly population densities in each plot were assessed every week by placing five Multilure<br />
baited trap (Better World) with three components food-based synthetic lure containing<br />
Ammonium Acetate, Putrescine and Trimethylamine (BioLure, Suterra LLC) and water plus<br />
Triton added to the base in order to retain attracted flies. The traps were sampled after 24 h and<br />
the number of flies captured was determined. The fruit was sampled once a week in all<br />
treatments in order to assess its infestation levels. One hundred fruits were sampled in each<br />
treatment and were dissected. The larvae found were put in Petri dish until adult emergence.<br />
The fruit infestation levels in each treatment are given.<br />
Results: In both tests the Mass Trapping Treatment (a) showed similar control and protection to<br />
the Ground Bait Treatment (Spinosad) (c) in fruit infestation levels and cached adults in<br />
Multilure traps. The Bait Station Treatment showed a good control in Test 1 (very closed to<br />
Ground Bait and Mass Trapping results) but in Test 2 its performance appeared to be not as<br />
good probably due to the exceptionally high populations of Medfly. Nevertheless, even under<br />
these unfavorable conditions the Bait Stations Treatment provided a remarkable control when<br />
compared with the untreated control.<br />
Conclusions: We consider that the three treatments protected the orange orchards during the<br />
entire time of fruit susceptibility. The Mass Trapping and Bait Station methods provide a<br />
useful medfly management tool which can be combined with other IPM methods used in the<br />
area. These procedures have the advantage of being environmentally friendly, not polluting the<br />
atmosphere and preserving the beneficial entomofauna. This technology is very important in<br />
integrated or organic fruit production, and for several markets in which the control of<br />
insecticide residue is important.<br />
Keywords: Medfly, bait station, alternative control, mass trapping, Fruit Fly SIT.<br />
307
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
ENRICHED GINGER OIL, A POWERFUL SEXUAL ATTRACTANT FOR TRAPPING THE<br />
MALES OF SOME CERATITIS SPP.<br />
Quilici, Serge* 1 ; Schmitt, Chloé 1 ; Trelluyer, Mickael 1 ; Franck, Antoine 1 ; Deguine, Jean-Philippe 1 &<br />
Peñarrubia-María, Esther 2<br />
1 CIRAD (Centre de Coopération Internationale en Recherche Agronomique pour le Développement),<br />
Département des Systèmes Biologiques, Saint-Pierre (La Réunion), France. E-mail: serge.quilici@cirad.fr 2<br />
Institut de Recerca i Tecnologia Agroalimentàries (IRTA) - Estació Experimental Agrícola Mas Badia. Canet de<br />
la Tallada, 17134 La Tallada d’Empordà, Girona, Spain.<br />
Background: Since the 60’s, trimedlure has been the main para-pheromone used to attract the<br />
males of a series of Ceratitis spp. for monitoring or detection purposes. In recent years, within<br />
a FAO-IAEA coordinated programme, in the framework of SIT (Sterile Insect Technique)<br />
programmes, the strong attractiveness of Ginger Root Oil (GRO) for the males of some<br />
Ceratitis spp. has been confirmed. Based on these results, it appeared useful to compare the<br />
attractiveness of a recently available commercial dispenser of Enriched Ginger Oil (Insect<br />
Science SA) with that of a standard trimedlure dispenser, in field trapping experiments. More<br />
recently, field-cage tests were carried out to determine the factors (age, food status, mating…)<br />
influencing the male response in both species.<br />
Method: Four field experiments were conducted in 2009-2010 on different crops. The first one<br />
was carried out in a Citrus orchard situated in Bassin Martin, in the south of the island, at an<br />
altitude of 290 m. The second trial was carried out in another Citrus orchard, in Petite Ile in the<br />
south (alt.: 300m) and the third one in Etang-Salé, in the south-west, in a chilli plot (alt.: 10m).<br />
Finally a fourth trial, set up in Piton Armand on the East in a Chinese guava plot (alt.: 450m),<br />
is still in progress. For each trial, two types of attractants were compared in Tephri-traps<br />
(Sorygar, Spain): Enriched Ginger Oil (EGO) (“Pherolure”, Insect Science SA, Tzaneen, South<br />
Africa) and trimedlure (Agrisense BCS Ltd, Pontypridd, UK). The attractant dispensers were<br />
placed in a small plastic basket at the top of the trap, while a strip of DDVP was placed at the<br />
bottom of the Tephri-trap to kill the adult flies. From four to six replicates were carried out<br />
depending on the trial. Traps were monitored weekly for recording the number of caught<br />
males, then rotated. For each species, results were analysed with a GLM (Poisson family, logit<br />
link) on the number of flies caught per trap, using the soft ware R.2.8.1.<br />
Result: In trials 1, 2 and 4, the populations of C. capitata were lower than those of C. rosa,<br />
while in trial 3, in the dry south-west of the island, the medfly dominated. In all trials (except<br />
trial 4, still in progress), for both species, EGO attracted highly significantly more males than<br />
did trimedlure (glm, 1 d.f., p
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
DESIGNING CONTROLLED RELEASE DEVICES: FORMULATING FRUIT FLY<br />
ATTRACTANTS AND COMMERCIALLY AVAILABLE INSECTICIDES.<br />
Ramsey, Amanda* 1 & Jang, Eric 2<br />
1 Scentry Biologicals, Inc. 610 Central Avenue, Billings, MT, USA; Email: aramsey@scentry.com; 2 USDA-ARS,<br />
Pacific Basin Agricultural Research Center, POB 4459, Hilo, HI 96720 USA<br />
Background: The development of controlled release devices (CRDs) can be summarized as<br />
being dictated by the following factors: (1) the chemistry of the semiochemical (2) the targeted<br />
level of attractancy (3) economics. The design of the CRD will account for the volatility and<br />
solubility of the attractant, for example equal quantities of methyl eugenol and cue lure will not<br />
have the same longevity. To obtain equal longevities, formulations for methyl eugenol have a<br />
higher loading, and a different ratio of vinyl polymer to compensate for the increased volatility<br />
of the molecule. Furthermore, formulations for cue lure are developed to not last beyond the<br />
required field-life, thus preventing residual attractant from remaining within the CRD. In<br />
summary, semiochemicals are formulated to achieve optimal release from polymer-based<br />
CRDs. A general practice of monitoring fruit flies in the United States, specifically regions<br />
such as California and Florida, entails absorbing an attractant and an insecticide onto a cotton<br />
wick and positioning the combination into a Jackson Trap with a sticky bottom. The<br />
preparation of the cotton wick is laborious and the users are exposed to liquid insecticides such<br />
as Naled (Dimethyl 1,2-dibromo-2,2-dichloroethyl phosphate) and DDVP (Dimethyl 2,2dichlorovinyl<br />
phosphate). Combining the attractant and the insecticide within a polymeric CRD<br />
would minimize exposure to insecticides and eliminate the added task of baiting wicks.<br />
Methods: A study was conducted on native populations of Bactrocera dorsalis, Oriental Fruit<br />
Fly, on the Big Island of Hawai’i. The 8-week trial compared baited wicks and the Scentry<br />
Methyl Eugenol Cone combined with the following commercially available insecticides:<br />
Naled, DDVP, Spinosad, and Rynaxypyr. The lures and wicks were baited with 6 ml of<br />
attractant and 5% of the insecticide.<br />
Results: The DuPont Rynaxypyr formulation demonstrated strong competitive insecticide<br />
activity to the organophosphate formulations at a smaller percentage or loading of insecticide<br />
per device.<br />
Conclusion: Every one of the all-in-one products present in the study were competitive to the<br />
loaded wicks, offering field technicians a viable alternative to the old method of baiting cotton<br />
wicks.<br />
Keywords: Bactrocera, Methyl Eugenol, Cue Lure, Rynaxypyr.<br />
309
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
COMPARISON OF EFFICACY OF BACTROGEL AND SUCCESS APPAT FOR THE<br />
CONTROL OF CUCURBIT FRUIT FLY, DACUS DEMMEREZI, IN MADAGASCAR.<br />
Raoelijaona, Jeannette C. Y. 1 ; Raoelijaona, Adrien R. 1 ; Raharimalala, Soanjanahary 1 ;<br />
Ratovonomenjanahary, Zelin T. 2 ; Rasamizafy, Lerry A. 2 & Andrianarimanana, Aina G. 3.<br />
1 Ministère de l’Agriculture, Direction de la Protection des Végétaux, Division Entomologie, Antananarivo,<br />
Madagascar. Email: raoelijaonaj@yahoo.fr; 2 Université d’Antananarivo, Faculté des Sciences, Département<br />
Entomologie, Antananarivo,Madagascar; 3 Groupe Agriculture Chimie Madagascar (ACM), Antananarivo,<br />
Madagascar.<br />
Background: Cucurbit production is one of producer’s income sources. Unfortunately, this<br />
culture either cultivated or wild is submitted to Dacus demmerezi, the cucurbits fruit flies<br />
attacks. The harvest lost could be assessing at 85%. In order to implement a survey strategy<br />
against this pest the team thinks that it is judicious to find an effective product which would be<br />
advised to the producers<br />
Methods: All experiences have been lead on the ACM Society parcel. Maize has been growth<br />
at the four parcels corner and Mac Phail trap with cue lure has been placed in the center. Every<br />
fifteen days, captured flies in the trap has been counted and maize has received application<br />
with either Bactrogel or Success Appat<br />
Results: The graph of fruit flies population evaluation has shown that Dacus has a good<br />
response as well to Bactrogel as to success appat but there is little bit difference on capture.<br />
Conclusion: We conclude that Success Appat is more effective than Bactrogel<br />
Keywords: Success Appat, Dacus, Bactrogel
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
MASS TRAPPING OF CERATITIS CAPITATA AND NON-TARGET CONTROL FLIES.<br />
Reuveny, Haim<br />
Integrated Pest Management Center (IPMC), Northern R&D. Israel<br />
Background: Control of fruit fly Ceratitis capitata (Diptera: Tephritidae), in Israeli apple<br />
orchards is based on airborne, wide-area, ULV spraying with malathion mixed with protein<br />
hydrolysate (PH). In IPM orchards, where use of malathion and other organophosphates is not<br />
advisable, attractant-formulation devices are used, without pesticides (Cera Trap, Olipe) or<br />
with a selective pesticide (spinosad) (Biofeed, Frutect06). This study examined attraction of<br />
flies, including C. capitata (male and female) and non-target control flies to such devices and<br />
systems.<br />
Methods: The first trial, in four commercial apple orchards (each about 2 ha) in northern Israel,<br />
used four different attractant devices to control C. capitata: 1) Cera Trap at 100/ha; 2) Olipe,<br />
100/ha; 3) Biofeed, 150/ha; 4) Frutect06, 150/ha. The first two are non-toxic devices based on<br />
attractant formulations of PH and Torula yeast, respectively. Attraction to the last two is based<br />
on an odorous mixture of pesticide (PH and spinosad), in combination with color and form (a<br />
square yellow board with a red circle). Six such devices were placed randomly in each<br />
treatment orchard after some modifications to facilitate fly trapping: 1) The Cera Trap and<br />
Olipe bottle were cut off below the fly entry hole. To avoid drowning the flies the piece of net<br />
and the DDVP plug (the lethal component) where placed above the attractant formulation; 2)<br />
The Biofeed and Frutect06 were covered with transparent covers and sprayed with insect glue.<br />
The devices were checked every week from July through December. The second trial involved<br />
airborne wide area control in citrus orchards; the same attractant formulations were tested, all<br />
of them poured from bottles. The trial was arranged in five random blocks with 25 m between<br />
devices. The devices were checked weekly from January through April. To prevent possible<br />
influence of location of fly trapping (i.e., fly hot spot) the device was moved to the adjacent<br />
location within the same block after being checked.<br />
Results: Significantly more trapping of C. capitata in apple orchards (Jul.-Dec.) were found<br />
with Olipe (average of 25.7 per device) and Cera Trap (23.0) than with Frutect06 (5.5) and<br />
Biofeed (0.4); the male:female ratios were 1:4, 1:2, 1:1.7, and 1:1, respectively. All devices<br />
also trapped non-target flies (NTF) at average rates of 290.3, 310.0, 56.2 and 51.2 per device,<br />
respectively. The trapping ratios of C. capitata:NTF were: 1:11.3, 1:13.5, 1:10.2, 1:128.0,<br />
respectively. In citrus (Jan.-Apr.) C. capitata trappings averaged 1.4, 12.4, 2.8 and 9.4 per<br />
device in Olipe, Cera Trap, Frutct06 and Biofeed, respectively, with male:female trapping<br />
ratios of 1:6, 1:2.3, 1.1.8, 1:1.6, respectively. The average rates of NTF trapping were 488.8,<br />
898.0, 621.8 and 168.8 per device, respectively, and the C. capitata:NTF trapping ratios were:<br />
1:349.1, 1:72.4, 1:222.1, and 1:17.9, respectively.<br />
Conclusions: The results clearly show the problem presented by the higher rate of NTF<br />
trapping, with regard to an IPM strategy for controlling C. capitata if attractant devices using<br />
selective products are to be considered. The high rate of NTF and low rate of C. capitata<br />
trapping must be addressed in development of specific attractant formulations.<br />
Keywords: Mass trapping, Ceratitis capitata, Non-target flies.<br />
311
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
TRANSGENIC ORANGE FRUIT WITH REDUCED LIMONENE CONTENT IN THE PEEL<br />
IS LESS ATTRACTANT TO CERATITIS CAPITATA (WIEDEMANN).<br />
Rodríguez, Ana* 1 ; San Andrés, Victoria 1,2 ; Cervera, Magdalena 1 ; Shimada, Takehiko 3 ;<br />
Rodrigo, M.Jesús 4 ; Zacarías, Lorenzo 4 ; Castañera, Pedro 2 & Peña, Leandro 1<br />
1 Instituto Valenciano de Investigaciones Agrarias (IVIA), Centro de Protección Vegetal y Biotecnología. Ctra. de<br />
Moncada a Náquera km. 4,5; Moncada 46113- Valencia (Spain); Email: arodrig@ivia.es; 2 Unidad asociada UJI-<br />
CIB. Centro de Investigaciones Biológicas (CIB-CSIC), Departamento Biología de Plantas, Madrid, Spain.<br />
3 National Institute of Fruit Tree Science (NIFTS), Tsukuba, Japan; 4 Instituto de Agroquímica y Tecnología de<br />
Alimentos (IATA-CSIC), Valencia, Spain.<br />
Background: Ceratitis capitata (medfly) males form loose aggregations termed leks,<br />
performing sexual signaling by emitting a sexual pheromone. Receptive females visit leks and<br />
choose their mates on the basis of their physical characteristics and their courtship<br />
performance. The secondary compounds produced by plants, many of them are volatiles, have<br />
signaling functions. In Citrus, the peel is embedded of thousands of oil glands that contain<br />
terpene volatile compounds, being limonene the most abundant one (up to 97% in orange<br />
fruits). The high amount of limonene that accumulates in orange oil glands suggests an<br />
important biological role for this terpene compound in fruit odour interactions with the biotic<br />
environment. However, some studies have suggested mating advantages when adult males<br />
were exposed to different volatiles derived from orange fruits.<br />
Methods: Transgenic plants expressing the complete cDNA of a citrus limonene synthase gene<br />
(CiTMTSE1) in antisense orientation (AS) were used in this study. Pure synthetic terpene<br />
compounds were used in laboratory experiments. Neonate larvae and 5 day-old laboratory<br />
mass-reared flies were used in laboratory, greenhouse and wind tunnel bioassays.<br />
Results: Transgenic plants had a decrease of two orders in magnitude of limonene<br />
accumulation in fruit peel. Consequently, the accumulation of other monoterpenes and<br />
sesquiterpenes decreased while that of monoterpene alcohols increased. Wind tunnel bioassays<br />
with medfly males exposed to pure compounds revealed a clear attraction to linalool and<br />
limonene volatiles. In no-choice experiments, AS and control fruits were damaged at<br />
comparable levels by medfly females. Nevertheless, wind tunnel bioassays with AS and control<br />
fruits showed that males were not only not repelled but significantly more attracted to control<br />
fruits (more than ten times; 32% vs. 2%), indicating that limonene (and likely other terpene)<br />
emission were actually insect- attractants to the fruit.<br />
Conclusions: These results provide a much more comprehensive view of the role of terpene<br />
volatiles as attractors of medfly in Citrus fruits.<br />
Keywords: Limonene, volatiles, antisense downregulation, Ceratitis capitata behaviour
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
TRANSPARENT VERSUS YELLOW COLOR IN TRAPS AGAINST BACTROCERA OLEAE<br />
(ROSSI).<br />
Ros, J. Pedro* & Seris, Elena.<br />
Instituto Nacional Investigaciones Agrarias. Ctera Coruña Km.7 28040 Madrid. (Spain). Email: ros@inia.es<br />
Background : Currently, yellow is the most frequent color in traps against Tephritidae family.<br />
The first Macphail traps that began it way made with transparent glasses, today, because the<br />
manufacturing, transport and prices, have derived into plastics and yellow color as the<br />
reference. The fundament of Easy trap ® (Sorygar,Co) (as the main different with Mcphail) is to<br />
have two opposite “Ventury” holes that makes the vacuum inside the trap when the air is<br />
coming in, forcing out the smell of the bait when the air is coming out. Easy trap has a high<br />
efficiency degree when capturing fruit flies. It is working with solid bait (Biolure ® Suterra,Co)<br />
as well as liquid baits (Hydrolysate protein) but even though to be airtight the farmers don’t<br />
close it well and the liquid spill. A new version “Easy Blister Trap ® ” still keeping the Easy trap<br />
concept (ventury holes) but it is a completely transparent and completely sealed without any<br />
manual interaction. We have carried out an experiment to determine the variability of both<br />
traps capturing Bactrocera oleae and studying if it have differences on the attracted<br />
parasitoids and predators.<br />
Methods: A ecological olive farm of 4 ha. sited in Villarejo (Madrid Province) was chosen to<br />
hung the traps, Easy (yellow) and Easy Blister Trap (Transparent) both baited with Tephri<br />
Lure ® 12% from 10 June to 12 November of 2009 (165 days). The statistical design was 4<br />
replicates of both traps on alternate olives and consecutive rows separated each other (12x24<br />
m). The check was made each 15 days, counting males and females as well as other insects. A<br />
statistical analysis of variance was carried out with the data.<br />
Results: No significant differences (α = 0.05 ) between the captures of both traps (males and<br />
females) although in all checks the transparent trap capture more flies tan the yellow one.<br />
Regarding numbers of non-target arthropods captured, the percentages of natural enemies were<br />
6,1% for Easy trap and 5,6 for Easy Blister Trap but no statistical significant differences were<br />
observed between these numbers, either in the case of predators or parasitoids.<br />
Coclusions: According with the results; neither Bactrocera oleae, nor predators, nor parasitoids<br />
change their behavior when trap color is changed from yellow to transparent. This experiment<br />
is giving some evidences that the most important factors for trap designing is the kind of<br />
attractant and the way the trap spread it in the air and not the color of the trap<br />
Keywords: Bactrocera oleae, easy trap, easy blister trap, natural enemies<br />
313
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
AMMONIUM PHOSPHATE VERSUS HYDROLYSATE PROTEIN: WHICH ONE TO BE<br />
USED IN MASS TRAPPING AGAINST THE OLIVE FLY BACTROCERA OLEAE, (ROSSI).<br />
Ruiz, J. Manuel¹*; Ruiz, Joaquín²; Alonso, Esther³ & Ros, J. Pedro 4<br />
1 Laboratorio de Producción y Sanidad Vegetal. GEOLIT. Parque Científico y Tecnológico. C/ Sierra Morena,<br />
Manzana 12 b. 23620 Mengíbar, Jaén. España. Email: manuel.ruiz.torres.ext@juntadeandalucia.es; ²Delg. Prov.<br />
Agric. Almería. Dept. Sanidad Vegetal C/Hermanos Machado 4 pl.4ª 04004 Almería .España. ³A.T.R.I.A. Unión<br />
de Cooperativas de Madrid (UCAM). C/ Jordan 8 28010 Madrid, España. 4 Instituto Nacional de Investigaciones<br />
Agrarias. Laboratorio de Entomología Agroforestal. Departamento de Protección Vegetal. Carretera de La<br />
Coruña, Km 7,5. 28040 Madrid. España.<br />
Background: Mass trapping is one of the methods to control olive fly when insecticide is<br />
forbidden, case of organic grows. Olipe trap baited with Ammonium Phosphate is a wellknown<br />
trap extensively used in many countries. Many years ago we have demonstrate that<br />
hydrolysate protein is a better attractant than Ammonium Phosphate against olive fly (IAEA<br />
programmes). This method arise a high cost of money and hand work if we not guarantee the<br />
maximum of captures that makes a free damage harvest. The authors are working several years<br />
with a new protein hydrolysate (Tephri Lure ® , Sorygar,Co) needing to check that there are no<br />
errors. In this poster we present the results of four simultaneous trials carried out in four<br />
separated sites with very different climates.<br />
Methods: The experiment was carried out in four localities of Spain: Jaén (the greatest area of<br />
olive culture), Almería, near the Mediterranean sea, and Madrid (2), middle of Spain with<br />
continental climate. We have used two kind of traps: Olipe (water mineral bottle) with 4 holes<br />
(1,5 cm ǿ) and Easy Blister trap a new version of “Easy Trap ® ” Sorygar,Co) that keeping the<br />
Easy trap concept (ventury holes) but it is a completely transparent and completely sealed<br />
without any manual interaction. Each trap was baited with Ammonium Phosphate 4% and<br />
Tephri Lure 12% (from origin with borax and glycol). The traps were hung on the interior of<br />
the olive tree. Four replicates were placed randomly in a square surface of sixteen olive trees.<br />
(12X12m.). The check of traps was monthly, males and females were counted just like the<br />
different arthropods. The statistical analysis of variance was made with the change of variable<br />
x’=(x+1) 1/2<br />
Results: In all localities were the experiment was carried out the hydrolysate protein has<br />
captured much more flies independent of the type of trap. Easy Blister Trap was better than<br />
Olipe Trap when both trap are baiting with Ammonium Phosphate, but they are not significant<br />
differences when they are baiting with hydrolisate protein. (F=11,3; p
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
EFFECT OF SUBLETHAL CONCENTRATIONS OF NEEM EXTRACTS ON THE<br />
REPRODUCTION AND LONGEVITY OF THE MEDFLY, CERATITIS CAPITATA<br />
(DIPTERA: TEPHRITIDAE).<br />
Silva, Márcio Alves 1 ; Bezerra, Gerane C. Dias 1 ; Mastrangelo, Thiago* 2 & Vendramim,<br />
José Djair 1<br />
1 Department of Entomology and Acarology, University of São Paulo (USP). Av. Pádua Dias 11, 13418-900,<br />
Piracicaba, SP, Brazil. Email: marcio@esalq.usp.br; 2 Center for Nuclear Energy in Agriculture (CENA/USP).<br />
Av. Centenário 303, 13400-970, Piracicaba, SP, Brazil.<br />
Background: Fruit flies show high vitellogenesis at the adult stage, which represents a good<br />
moment to verify the physiological effects of neem extracts. The sublethal concentrations<br />
(LC15, LC30 and LC45) of aqueous extract from seed kernels of Azadirachta indica A. Juss for<br />
Ceratitis capitata (Wied.) adults were estimated applying bait (extract + honey) on ‘Cravo’<br />
lemon seedlings. Subsequently, the effect of different extracts in these sublethal concentrations<br />
(39, 225 e 888 ppm, respectively) on the longevity, fecundity and the fertility on the 1 st , 4 th , 8 th<br />
and the 12 th days of oviposition was evaluated.<br />
Methods: As first step, using the aqueous extract from neem seed kernels, the LC15, LC30 and<br />
LC45 were estimated for adults exposed to a bait (a solution of 20 % honey + neem) in cages<br />
containing a citrus seedling. After that, those concentrations were used for a second bioassay. It<br />
was used plastic cages where it was fixed a cotton wick which was in contact with a plastic vial<br />
with 12.5 ml of the extracts (seed kernel aqueous extract, leaf extract in methanol and branch<br />
extract in dichloromethane). Water or 5 % acetone were used as controls. Two couples were<br />
released in each cage and an artificial diet (hydrolyzed yeast + sugar) was offered ad libitum.<br />
The sublethal concentrations were estimated by probit regression analyses using the SAS 9.1<br />
software. The experiment was completely randomized with four replicates per treatment and<br />
was conducted under environmental controlled conditions (25 ± 2ºC, 60 ± 10 % RH and 14 h<br />
photophase).<br />
Results: At these sublethal concentrations, the longevity of males and females and the preoviposition<br />
period were not significantly affected. The branches extract in dichloromethane at<br />
the highest concentration (888 ppm) affected significantly the fecundity and the fertility at the<br />
8 th day, reducing in almost 80 % the number of eggs laid and in 30 % the egg hatching.<br />
Conclusions: The estimated LC15, LC30 and LC45 were 38.68, 225.19 and 887.86 ppm for the<br />
aqueous extract of neem seed kernels in the bait. The branches extract in dichloromethane at<br />
the concentration of 888 ppm affected significantly the insect fecundity and fertility.<br />
Keywords: Medfly, Azadirachta indica, sublethal concentrations, reproduction.<br />
315
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
EFFECT OF SPINOSAD, KAOLIN AND PROTEIN BAIT SPRAY ON CERATITIS CAPITATA<br />
(DIPTERA: TEPHRITIDAE) POPULATION IN CITRUS ORCHARDS.<br />
Smaili, Charif 1 ; Wadjinny, Jamila 2 & Bakri, Abdeljelil 3*<br />
1 INRA, Laboratory of Entomology, Kenitra, Morocco; Email: csmaili@yahoo.fr; 2 IRPV, ONSSA, Kenitra,<br />
Morocco; 3 Insecte Contrôle, Marrakech 40080, Morocco.<br />
Background: Phytosanitary practices based on the extensive use of insecticides in citrus groves<br />
to control the Mediterranean fruit fly Ceratitis capitata (Wiedemann) may cause the resurgence<br />
of resistant strains of this pest, the emergence of secondary pests by elimination of their natural<br />
enemies, and the growing public concerns over issues related to public health, environmental<br />
quality, and food safety. Therefore, there is an urgent need to develop an alternative and<br />
environment friendly method for C. capitata management. The purpose of this study was to<br />
demonstrate the efficiency of the treatment with the kaolin, spinosad and protein bait spray<br />
(BAT) and how these different treatments may be associated for a better control of C. capitata<br />
in several citrus varieties.<br />
Methods: Citrus plots of Clementine (Citrus reticulata, var. Cadoux), sweet orange (Citrus<br />
sinensis, var. Hamlin), sweet orange (Citrus sinensis, var. WashingtonNavel), and Clementine<br />
Mandarin (Citrus reticulata, var. Marisol) were treated with kaolin for 7 days at two doses.<br />
Results & Conclusions: The plots treated with the kaolin continuously for seven days showed a<br />
lower number of C. capitata captures and fruit infestation compared with those treated with<br />
BAT or spinosad. Kaolin used at a low dose (1.4%) showed a similar level of infestation as<br />
spinosad. However, at a high dose (3%), a small reduction in fruit infestation is obtained<br />
compared to spinosad. Addition of the hydrolyzed protein does not seem to enhance spinosad<br />
effectiveness. The feasibility of combining resistant citrus varieties with these alternative<br />
methods is discussed and would be considered in the development of an integrated pest<br />
management strategy against medfly in citrus groves.<br />
Keywords: Citrus, Ceratitis capitata, Kaolin, Spinosad, Pest management.
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
MONITORING RHAGOLETIS ALTERNATA FALLEN – A SERIOUS PEST OF ROSE HIPS.<br />
Uggla, Madeleine<br />
Balsgård-Department of Plant Breeding and Biotechnology, Swedish University of Agricultural Sciences,<br />
Fjälkestadsvägen 459, SE-Kristianstad, Sweden. E-mail: madeleine.uggla@ltj.slu.se<br />
Background: Due to their biochemical compounds the interest of growing rosehips, the fruits<br />
from different Rosa species (mainly section Caninae), has increased the last years. In orchards<br />
growing raw material to health promoting products there is a demand for organic production.<br />
Rhagoletis alternata is the most serious pest in wild-growing as well in cultivated rosehips.<br />
From a previous study, a high percentage of the wild-growing plants had infested fruits. Initial<br />
study of phenology of R. alternata has been performed.<br />
Methods: Rhagoletis alternata was studied during three seasons in an orchard in the south of<br />
Sweden. The plant material originated from a plant breeding program and included several<br />
Rosa species. Using three-dimensional unbaited yellow sticky traps (Rebell amarillo) adult<br />
flies were monitored from June to September. The weather data was collected and the degreedays<br />
have been calculated.<br />
Results: In 2007 more flies were captured on traps compared to the same period in 2008 and<br />
2009. In all three years the peak of fly capture occurred at the same time on calendar basis.<br />
Differences in captures between traps showed possible genetic variation in Rosa species in<br />
their chemical content.<br />
Conclusions: The study demonstrated that the Rebell amarillo was effective for capturing R.<br />
alternata flies. Further study will probably show interaction between the pest and different<br />
Rosa genotypes.<br />
Keywords: Rosehip fly, traps.<br />
317
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
USE OF FOLIAGE BAIT SPRAYS IN THE REDUCTION OF BACTROCERA INVADENS<br />
(DREW, TSURATA AND WHITE) DAMAGE ON SWEET ORANGE VARIETIES.<br />
Umeh,Vincent*; Olaniyi, Mercy & Thomas, John<br />
National Horticultural Research Institute (NIHORT), Citrus Research Program, P.M.B. 5432 Idi-Ishin, Jericho<br />
Reservation Area, Ibadan, Nigeria, E-mail vumeha@yahoo.com<br />
Background: Damages caused by fruit flies have been identified as major limiting factors to<br />
citrus production in Nigeria. Recently, a newly introduced species Bactrocera invadens caused<br />
economic yield losses in many producing areas. The majority of sweet orange producers in<br />
Nigeria are small-holders who do not often adopt standard citrus production practices, usually<br />
due to lack of the necessary funds. The present study therefore seeks to evaluate the application<br />
of locally-made protein bait sprays in different sweet orange varieties in the reduction of fruit<br />
fly damage.<br />
Methods: In 2006 and 2007, one, two and three spot applications of locally-made protein bait<br />
mixed with chlorpyrifos insecticide were made separately to the foliage of Agege, Valencia<br />
Late and Parson Brown sweet oranges arranged in randomized complete block and replicated<br />
thrice at the National Horticultural Research Institute, Ibadan. Sweet orange stands with no<br />
treatments were maintained as control. The candidate trees were picked at random. Efficacy of<br />
the bait sprays in reducing fruit attack was assessed by randomly sampling 10 ripe or ripening<br />
fruits per stand. Each spot spray covered 50cm x 50cm of the foliage. Sweet orange stands<br />
with no treatments were used as controls. Efficacy of the bait sprays in reducing fruit attack<br />
was assessed by close observation of fruit fly entry spots (with the aid of hand lens) in 10 ripe<br />
or ripening fruits randomly sampled per stand. The number of fruits that dropped underneath<br />
each candidate tree due to fruit fly damage was recorded. The fruits were picked and buried<br />
away from the site. Confirmation of fruit fly species causing sweet orange damage was<br />
achieved by rearing fruit flies in attacked fruits placed inside wooden cages till adult<br />
emergence. Data were subjected to Analysis of Variance (ANOVA) procedures and means of<br />
significant tests were separated using Student-Newman-Keuls test at P≤ 0.05.<br />
Results: The bait sprays significantly (P
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
IMPACT OF WEAVER ANTS (HYMENOPTERA: FORMICIDAE) ON MANGO FRUIT<br />
FLY (DIPTERA: TEPHRITIDAE) DAMAGES IN BENIN IN 2008-2009.<br />
Vayssières, Jean-François* 1 ; Sinzogan, Antonio 1 ; Ouagoussounon, Issa 1 ; Adandonon,<br />
Appolinaire 1 & Van Mele, Paul 2<br />
1 International Institute of Tropical Agriculture, Centre International de Recherche (IITA-CIRAD), 08 BP 0932,<br />
Cotonou, Benin. Email: j.vayssieres@cgiar.org; 2 Africa Rice Center (ARC), Cotonou, Benin<br />
Background: In sub Saharan Africa the mango fruit industry is facing with the problem of fruit<br />
flies. Among the numerous species that attack the mango, a new invasive species, Bactrocera<br />
invadens, is causing especially extensive damage compared to the native fly species during the<br />
rainy season. It has a number of comparative advantages such as good flying skills and<br />
competitive demographic parameters which allow it to re-infest orchards relatively quickly.<br />
Making more efficient use of natural enemies can be of great benefit to the growers. As Asian<br />
growers are using Asian weaver ant (Oecophylla smaragdina) since many centuries it made<br />
sense to test the ability of African weaver ant (Oecophylla longinoda) to be an effective<br />
biological control agent in Africa. We have studied the ant impact on early cultivar (cv) in<br />
2006-2007. It was interesting to do the same kind of experiment on season and late cv.<br />
Methods: In Benin, we tested the impact of weaver ants on both B. invadens and Ceratitis<br />
cosyra on 5 mango cv (Ifac 3, Améliorée du Cameroun, Kent, Smith and Brooks). The first<br />
three ones are season cv and the last two ones are late cv. We used three blocks in 2008 (Ifac 3,<br />
Kent, Smith) and three other blocks in 2009 (Améliorée, Smith and Brooks) with two<br />
treatments each (i) without ants = control, (ii) with ants. Each treatment was repeated 10 times.<br />
Data were analysed by one-way ANOVA followed by Student Newman Keuls test. Analysis<br />
was done using the SAS Statistical Program(SAS 2003).<br />
Results: Mean damage inflicted by fruit flies was significantly reduced by 58% in 2008 on Ifac<br />
3, Kent and Smith between mango trees with ants vs control trees. In 2009, mean damage<br />
inflicted by fruit flies was significantly reduced by 81% in other blocks of Améliorée du<br />
Cameroun, Smith and Brooks. The control mangoes cv Kent, Smith and Brooks had the highest<br />
infestation with B. invadens populations. It is normal because a consistent population increase<br />
of B. invadens in the early rainy season caused considerable damage to mid season (Kent) and<br />
late cultivars (Smith, Brooks).<br />
Conclusions: The use of Oecophylla colonies is well-suited for perennial cropping systems in<br />
sub Saharan Africa, because they are efficient, constantly available, widespread and selfregenerating.<br />
Practical information about the use of weaver ants in fruit fly control should be<br />
made available to all those involved in the mango fruit industry at every level.<br />
Keywords: conservation biological control, Mangifera indica, fruit flies, West Africa.<br />
319
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
THE STUDY ON THE APPLICATION METHODS OF PROTEIN BAIT TO ORENTIAL<br />
FRUIT FLY.<br />
Wang, Bo 1 & Chen, Jia-hua 2<br />
Beneficial Insects Institute, Fujian Agriculutre and Forestry University, Fuzhou, China. Email:<br />
chenjh34@yahoo.com.cn.<br />
Background: Oriental fruit fly, Bactrocera dorsalis (Hendel), is a tephritid fruit fly that is a<br />
serious pest of a wide range of tropical fruits. Application protein bait was one important<br />
measure to prevention and control B. dorsalis (Hendel). Using trap bottle and spot spray were<br />
two important means of application protein bait.<br />
Methods: The effect some factors to attractant effectiveness were studied in this paper, i.e<br />
using methods, amount and cycle of protein bait,and comparing the application effect with<br />
different host orchards, different protein bait and different locations.<br />
Results: The effect of diluting two-fold was best regardless of means, and the number of<br />
attractant and deadth were 5.33 and 6.67 respectively. The optimal use amount of two methods<br />
were 15mL and 30mL, even though the attractant effectiveness increased with the increase of<br />
using amount. The life cycle of two means were the same as 5 days, and the the number of<br />
attractant and deadth were 12.33 and 13.33 on the first two days. The results of using trap<br />
bottle founded that the attractant effectiveness can be increased by raising pH value properly,<br />
and the best value of pH was 6.The attractant effectiveness in star fruit orchard was better than<br />
oranges and grapefruit orchards, and the atttractant number was 5.00. There was no significant<br />
difference between home-made protein bait and Prima, the attractant number were 7.33 and<br />
9.33 respectively, which significantly higher than GF-120. The attractant number of pake,<br />
orchard and campuses were 11.73, 7.00, and 5.33 respectively. The results of spot spray<br />
showed that the effect was best when adding 0.20% amount of malathion, and the adults<br />
number of deaths was 7.67.<br />
Conclusions: The methods have some benefits of guiding on field management.<br />
Keywords: protein bait; Bactrocera dorsalis (Hendel); application method; trap bottle; spot spray
Workshop<br />
Longevity, Ageing & SIT
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
TRANSCRIPTIONAL CHANGES RELATED TO MATURATION AND MATING IN MALE<br />
AND FEMALE CERATITIS CAPITATA WIEDEMANN.<br />
Gomulski, Ludvik M. 1 ; Dimopoulos, George 2 ; Xi, Zhiyong 2 ; Gabrieli, Paolo 1 ; Scolari,<br />
Francesca 1 ; Siciliano, Paolo 1 ; Malacrida, Anna R 1* & Gasperi, Giuliano 1<br />
1 University of Pavia, Department of Animal Biology, Via Ferrata 1, 27100, Pavia, Italy; Email:<br />
malacrid@unipv.it; 2 Johns Hopkins Bloomberg School of Public Health, Baltimore, USA<br />
Background: The medfly, Ceratitis capitata, is a highly invasive agricultural pest that has become<br />
a model insect for the development of biological control strategies. These control strategies<br />
are reliant on knowledge of the behaviour, physiology and molecular genetics of medfly<br />
reproduction. The recent availability of expressed sequence tags derived from adult medfly<br />
heads and embryos has permitted the development of microarrays for high throughput gene<br />
expression profiling. The genes that show changes in expression as a result of maturation and<br />
mating could potentially be used as targets for the development of novel medfly control strategies.<br />
Methods: A high density oligonucleotide microarray was designed based on 11885 assembled<br />
expressed sequence tags from medfly cDNA libraries derived from 0 - 8 day old male and female<br />
heads and from 0 - 36 h embryos. Complementary RNA derived from virgin immature,<br />
mature and mated males and females of the ISPRA strain were labelled and hybridised to the<br />
microarrays using standard Agilent protocols. The microarray data were filtered, normalized<br />
and analysed using TIGR MIDAS, MeV and GEPAS software. Gene Ontology and GO Term<br />
Enrichment analysis analyses were performed using GO Slimmer and GOEAST, respectively.<br />
The microarray-derived gene expression results were confirmed by comparison with the expression<br />
patterns of 12 genes obtained using real-time quantitative PCR (qRT-PCR). Three<br />
reference genes were used for relative quantification normalization.<br />
Results: A microarray based gene expression approach was used to compare the head transcriptomes<br />
of adult male and female medfly at differential physiological stages, sexually immature<br />
and mature virgin individuals and mated individuals. Particular attention was focused on the<br />
expression patterns of transcripts belonging to reproduction, behaviour, sensory perception of<br />
chemical stimulus, and immune system processes. Compared to the broad transcriptional<br />
changes during the maturation of the female, post-mating changes in females were modest,<br />
suggesting that in the medfly mating does not trigger extensive transcriptional changes in the<br />
head. Of particular interest is the apparent lack of mating-induced immune responses in the<br />
medfly female, compared to D. melanogaster, which may be the result of the different reproductive<br />
strategies of these species.<br />
Conclusions: Our study has revealed intriguing transcriptional changes during medfly maturation<br />
and mating. This study will encourage further investigations on numerous questions regarding<br />
the biology of this and related species. Apart from increasing our understanding of the<br />
molecular machinery behind these biological processes, the implicated genes may represent<br />
important targets that could be used to control populations of this pest species.<br />
Keywords: Ceratitis capitata, microarray, immune response, reproduction, gene expression<br />
323
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
GENOME-WIDE TRANSCRIPT PROFILES TO UNVEIL CERATITIS CAPITATA (WIE-<br />
DEMANN) CANDIDATE GENES INVOLVED IN AGEING.<br />
San Andrés, Victoria 1,2 ; Castañera, Pedro 2 & Sabater-Muñoz, Beatriz 1*<br />
Unidad Asociada de Entomología IVIA– CIB CSIC. 1 Instituto Valenciano de Investigaciones Agrarias (IVIA),<br />
Centro PVyB. Ctra. de Moncada a Náquera km. 4,5; Moncada 46113- Valencia (Spain). Email:<br />
sabater_bea@gva.es. 2 Centro de Investigaciones Biológicas (CIB) del Consejo Superior de Investigaciones<br />
Científicas (CSIC), Departamento Biología de Plantas, Madrid, Spain.<br />
Background: The success of a SIT program depends greatly upon releasing males at optimal<br />
sexual maturity and that they live long enough to successful mating with wild females.<br />
Consequently, information the knowledge of wild female reproductive status (mature or not) at<br />
the time of the sterile male release is a factor to be considered for the success of the SIT.<br />
Ageing is one of the most complex biological processes being determined by both, genetic and<br />
environmental factors. In C. capitata sex-specific life span patterns have been described as<br />
resulting from an underlying “constitutional” longevity minus the deleterious effects of<br />
reproductive biology, and sex-related behaviour. The role of genes in determining the rate of<br />
ageing and the primary mechanism underlying the ageing process are the subject of intense<br />
investigation. In recent years, sequencing analysis has become an important tool in molecular<br />
biology.<br />
Methods: In this work we present a genome-wide expression analysis based on Suppressive<br />
Substractive Hybridization technique (SSH) and Expressed Sequence Tag (EST) sequencing<br />
and macroarray expression analysis to identify signature genes related to the ageing-maturing<br />
process in C. capitata. Three differentiated reproductive (immature, mature and old adults)<br />
times have been used to perform SSH in circular scheme, both to females and males. After<br />
RNA extraction and SSH, all the substracted cDNA were cloned and single-pass sequenced.<br />
After comparisons to Genebank, consensus sequences were printed in a membrane and<br />
expression was measured by macroarray hybridization in each select time and compared in<br />
between by using GEPAS software.<br />
Results: We have sorted out 94 unigenes from 873 single-pass ESTs, of which 57% have<br />
homology with known genes. Ageing-maturing process in C. capitata presents a marked<br />
expression pattern accompanied by the increase of transcription level of genes involved in<br />
reproduction. In females, the maximum peak of expression (for vitellogenins and chorion<br />
proteins) is obtained at 5 days-old, whereas in males it is reached at the age of 15 days (for the<br />
male specific serum proteins). Other identified cDNAs with a differential expression pattern<br />
would be also candidates but deserves further studies, as they belong to the 43% unknown<br />
function class.<br />
Conclusions: These genes and the new candidates would allow proposing a transcriptional<br />
profiling protocol to determine the reproductive status and putative age of medflies in field.<br />
Keywords: Medfly, ageing, SSH, ESTs sequencing
AUTHOR INDEX
A<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
Abad, María ..................................................... 107, 266<br />
Aboussaid, Houda .................................................... 260<br />
Abraham, Solana .................................................. 52, 86<br />
Acín, Patricia ........................................................... 273<br />
Adán, Angeles .......................................................... 143<br />
Adandonon, Appolinaire .................................. 110, 319<br />
Adnan H. Salman .................................................... 294<br />
Afreh-Nuamah, Kwame........................................... 251<br />
Ahmad, Sohel........................................................... 219<br />
Ahmed, R.F. ............................................................. 220<br />
Alburqueque, Mónica ....................................... 147, 154<br />
Alcaide, Juan ........................................................... 160<br />
Alemansoor, H. ........................................................ 153<br />
Alfaro Cañamás, Cristina ......................... 126, 274, 301<br />
Ali K. Shbar ............................................................. 294<br />
Ali, Ali...................................................................... 249<br />
Alleck, Malini .......................................................... 245<br />
Almeida, Aparecida M. de ................................ 170, 212<br />
Alonso, Esther.......................................................... 314<br />
Alonso, Marina ........................................................ 253<br />
Alonso-Muñoz, Andrés ............................................ 275<br />
Alphey, Luke ............................................................. 66<br />
Al-Shammary, A.J. .................................................. 220<br />
Altamirano, Leopoldo .............................................. 135<br />
Al-Taweel, Ayad A................................................... 220<br />
Aluja, Martin .............................................. 62, 114, 268<br />
Alvarado, Ricardo ..................................................... 99<br />
Amala, U. ................................................................. 250<br />
Amornsak, Weerawan ......................... 50, 165, 246, 304<br />
Andrianarimanana, Aina G, .................................... 310<br />
Ant, Thomas H. ......................................................... 66<br />
Appiah, Ernest F...................................................... 251<br />
Araujo, Elton L......................................... 166, 167, 222<br />
Argilés, Rafael .................................................. 226, 291<br />
Arief, M. Ardelia ..................................................... 172<br />
Arnolds, Luciano ..................................................... 221<br />
Arouri, Rabeh ................................................... 184, 276<br />
Arques, Eva ...................................................... 206, 207<br />
Arthofer, Wolfgang ............................................ 47, 164<br />
Artiaga, Trinidad....................................................... 84<br />
Asimakopoulou, Anastasia K................................... 183<br />
Asís, José D. ............................................................. 253<br />
Atiama, Toulassi ................................ 131, 148, 150, 151<br />
Augustinos, Antonios ........................................ 183, 197<br />
Avilla, Jesús ............................................................. 306<br />
Ayala, Ildefonso ................................................ 107, 266<br />
Azandémé, Ginette .................................................. 123<br />
Azeredo-Espin, Ana M. L. ....................................... 244<br />
Azevedo-Filho, Joaquim A. de .......................... 170, 212<br />
B<br />
Bachmann, Guillermo .................................86, 152, 252<br />
Bachrouch, Olfa ....................................................... 277<br />
Backeljau, Thierry .....................................................70<br />
Báez M., Raúl ........................................................... 224<br />
Baeza Gómez, Jenny M. ........................................... 256<br />
Bakri, Abdeljelil ............................................... 144, 316<br />
Balagawi, Solomon .....................................................55<br />
Balmès, Valérie ......................................................... 203<br />
Banos, C.................................................................... 111<br />
Bargues, Laura ......................................................... 160<br />
Barnes, Brian.................................................... 204, 221<br />
Barr, Norman ................................................... 187, 194<br />
Barrón, V. ................................................................. 263<br />
Bartolucci, Andrea .....................................................61<br />
Batllori, Lluís............................................................ 306<br />
Beitia, Francisco ................................ 253, 260, 277, 278<br />
Belhadj Omrane ....................................................... 261<br />
Beltrán, Alina ........................................................... 240<br />
Benavides, Pablo.........................................................53<br />
Benevides, Clicia M. J. ............................................. 232<br />
Benevides, Leandro J. .............................................. 232<br />
Bengochea, Paloma ................................................... 305<br />
Bento, Albino A. ............................................... 143, 282<br />
Bento, Jose Mauricio S ............................................. 229<br />
Beroiz, Beatriz .................................................. 185, 276<br />
Bertin, Sabrina ...........................................................65<br />
Bezerra, Gerane C. D. .............................................. 315<br />
Biosca, E G. .............................................................. 211<br />
Bjelis, Mario ............................................................. 239<br />
Böckmann, Elias ....................................................... 253<br />
Boff, Mari Ines Carissimi ................................. 279, 280<br />
Bokonon-Ganta, Aimé .............................................. 287<br />
Bonomi, Angelica...................................................... 196<br />
Borges, Carlos A. ........................................................97<br />
Borges, Mirtha ......................................................... 240<br />
Borges, Rafael.................................... 127, 279, 280, 296<br />
Bortoli, Ligia Caroline.............................................. 280<br />
Botta, Anna .............................................................. 124<br />
Botton, Marcos ................................................. 279, 280<br />
Bourtzis, Kostas ........................................................ 183<br />
Bouvet, Juan P.......................................................... 152<br />
Braga Sobrinho, Raimundo...................................... 222<br />
Bravo, Bigail ............................................................. 233<br />
Bravo, J. .....................................................................89<br />
Breman, Floris .......................................................... 177<br />
Bressa, María José.................................................... 254<br />
Brévault, Thierry...................................................... 131<br />
Brohmsubha, Suttipun ...............................................88<br />
Brown, Leslie ............................................................ 213<br />
Budia, Flor................................................................ 143<br />
Buldawoo, Indrani .................................................... 245<br />
327
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Bull, Richard M ....................................................... 119<br />
Büttner, Carmen ...................................................... 249<br />
C<br />
Cáceres, Carlos ............................................. 81, 87, 219<br />
Caixeta, Claudini V. D. ..................................... 201, 202<br />
Callejas, Carmen ................................................72, 192<br />
Campos, José M. ............................................... 149, 160<br />
Campos, José Miguel ............................................... 297<br />
Campos, Mercedes ................................................... 143<br />
Canal, Nelson A. ...................................................... 178<br />
Cancino, Jorge ......................................................... 113<br />
Carabajal Paladino, Leonela ................ 48, 86, 252, 254<br />
Carbonell, Emilio ...................................... 231, 260, 278<br />
Cárdenas-Lozano, Jesús .......................................... 241<br />
Carey, James ............................................................ 125<br />
Carey, James R. ........................................ 135, 136, 137<br />
Casagrande, Enzo .................................................... 129<br />
Castañeda, Maria del R. .......................................... 178<br />
Castañera, Pedro 106, 107, 109, 184, 185, 226, 231, 265,<br />
266, 276, 312, 324<br />
Castro, Rosemary M. ..........................................90, 229<br />
Catalán-Ruescas, Diana ......................................77, 205<br />
Cervera, Amelia ................................................ 184, 185<br />
Cervera, Magdalena ................................................ 312<br />
Chandra Mouli, Munikote R. .................................. 243<br />
Chang, Chiou L. ....................................................... 233<br />
Chatterjee, Hirak ..................................................... 281<br />
Chen, Chia-Pang ...................................................... 121<br />
Chen, Chien-Chung ................................................. 121<br />
Chen, Jia-hua ........................................................... 320<br />
Chen, Yen-Hou .......................................................... 59<br />
Cheng, Edward Y..................................................... 242<br />
Chermiti, Brahim .............................................. 260, 277<br />
Chiang, Ming-Yao............................................. 101, 242<br />
Chinajariyawong, Anuchit ......................................... 56<br />
Chinvinijkul, Suksom ..........................88, 223, 230, 235<br />
Chiroleu, Frédéric.................................................... 131<br />
Choolun, Tejanand .................................................. 245<br />
Cicero Jarado, Lizzette ............................................ 268<br />
Cladera, Jorge L. ...... 48, 52, 86, 152, 236, 252, 254, 255<br />
Clarke, Anthony R. .......................... 55, 60, 91, 165, 209<br />
Cobo, Ana ................................................................ 305<br />
Cobos, Guillermo ..................................................... 305<br />
Cobos-Suárez, José M. ........................................77, 205<br />
Coelho, Mello A ....................................................... 229<br />
Coelho, Valentim ..................................................... 282<br />
Coltell, Néstor .......................................................... 283<br />
Conte, Claudia .................................................. 252, 255<br />
Costa, Karen .............................................................. 90<br />
Costa, Maria de Lourdes Z ........................................ 90<br />
Cotés, Belén.............................................................. 143<br />
Couso-Ferrer, Francisco ................................... 184, 185<br />
D<br />
Dabiré A Rémy ........................................................ 162<br />
Daneel, John-Henry ................................................. 128<br />
Dantas, Luis ............................................................. 208<br />
Davies, Andrew P. .................................................... 114<br />
de Alfonso, Ignacio................................................... 120<br />
De León-Crisóstomo, Ángel H. ........................... 85, 174<br />
de los Santos, Martín .......................................... 99, 241<br />
De Meyer, Marc .......... 42, 69, 70, 96, 155, 177, 194, 300<br />
De Siqueira Dutra, Vivian ......................................... 180<br />
de Souza-Filho, Miguel F. .......... 157, 158, 163, 170, 212<br />
De Villiers, Marelize .................................................. 76<br />
Deguine, Jean-Philippe . 49, 131, 148, 150, 151, 186, 308<br />
Delatte, Hélène ............................... 49, 70, 131, 150, 186<br />
Deus, Ezequiel G. .............................................. 168, 169<br />
Devescovi, Francisco ............................. 48, 86, 152, 252<br />
Diamantidis, Alexandros .................................. 136, 137<br />
Dias, Vanessa S. ....................................................... 232<br />
Díaz-Fleischer, Francisco ......................................... 228<br />
Dimopoulos, George................................................. 323<br />
Do Nascimento, Ruth R. .......................................... 173<br />
Domínguez, Julio .......................................... 84, 89, 224<br />
Dominiak, Bernard C. ............................................. 225<br />
Douraguia, Elisabeth ................................ 148, 150, 151<br />
Drew, Rai ................................................................... 56<br />
Drosopoulou, Eleni .................................................. 197<br />
Du Fretay, Gäel........................................................ 273<br />
Duyck, Pierre-François ............................................ 131<br />
E<br />
Egan, Mackenzie A. ................................................... 54<br />
Ekesi, Sunday......................... 42, 96, 112, 251, 262, 287<br />
El Messoussi, Said .................................................... 260<br />
El Meziane, Abdellatif ............................................. 258<br />
El-Heneidy, Ahmed H. ............................................. 115<br />
Encalada Mena, Lisandro A. ................................... 256<br />
Enkerlin, Walther ...................................................... 98<br />
Epsky, Nancy D......................................... 284, 295, 302<br />
Erbout, Nathalie ................................................. 69, 155<br />
Escudero-Colomar, Adriana ............................. 273, 306<br />
Espinosa, Eduardo ................................................... 283<br />
Espinoza, Hernán R. ................................................ 284<br />
F<br />
Falchetto, Marco ...................................................... 196<br />
Farfán, Fernando ....................................................... 53<br />
Farqad A. Sadq ........................................................ 294<br />
Farrar, Nasser.......................................................... 153<br />
Fay, Harry A. C. ...................................................... 122<br />
Feder, Jefferey. L ....................................................... 62
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
Fernandes, Thiago ................................................... 244<br />
Fernández Salinas, Maria L. ................................... 255<br />
Fernández, Milagros ......................................... 206, 207<br />
Fernández, Paz .......................................................... 72<br />
Fibla, José M............................................. 149, 160, 297<br />
Fischer, K ................................................................ 253<br />
Flores, S. .................................................................... 89<br />
Flores-Breceda, Salvador ................................. 257, 269<br />
Flores-García, Hector ................................................ 85<br />
Franck, Antoine ................................................ 131, 308<br />
François, Thomas .................................................... 151<br />
Frías, Daniel .............................................................. 61<br />
G<br />
Gabrieli, Paolo .................................................. 196, 323<br />
García Ramírez, María de Jesús ............................. 256<br />
García, Juliana ......................................... 147, 154, 227<br />
Garcia, Maria J. De M. ..................................... 170, 212<br />
Garcia-Marí, Ferran......................................... 160, 275<br />
García-Martínez, Víctor ............................. 85, 174, 188<br />
Garrido-Jurado, I. ................................................... 263<br />
Garziera, Luiza ................................................... 90, 229<br />
Gasperi, Giuliano................................... 65, 67, 196, 323<br />
Gava, Carlos .............................................................. 90<br />
Gayubo, Severiano F. .............................................. 253<br />
Geib, Scott ................................................................. 68<br />
Germain, Jean-François .......................................... 203<br />
Geurts, Katrien ........................................................ 155<br />
Gilchrist, Stuart ......................................................... 71<br />
Gilles, Benoît............................................................ 151<br />
Gil-Ortiz, Ricardo ................................................... 285<br />
Giraldo-Jaramillo, Marisol........................................ 53<br />
Girolami, Vincenzo .................................................. 195<br />
Gnanvossou, Desiré .......................................... 123, 287<br />
Gobet, Luis .............................................................. 210<br />
Godoy, Maria J. S. ................................................... 169<br />
Goergen, Georg ......................................................... 96<br />
Golestaneh, S.R........................................................ 153<br />
Gollahalli, Gundu R. V. ........................................... 243<br />
Gomes Silva, Janisete ................................................ 180<br />
Gomez, Luis E. ......................................... 127, 296, 298<br />
Gómez-Escobar, Enoc ............................................. 257<br />
Gomulski, Ludvik ....................................... 67, 196, 323<br />
Gonzales, Luis B. ....................................................... 75<br />
González, Manuel .................................................... 143<br />
Gonzalez, Ricardo ..................................................... 82<br />
González-Cabrera, Joel ........................................... 106<br />
González-Núñez, Manuel ......................................... 305<br />
Gourdon, Aude ........................................................ 203<br />
Griebeler, Eva M. .................................................... 187<br />
Griffin, Robert L. ...................................................... 75<br />
Guglielmino, Carmela R. ......................................... 196<br />
Guillén, Larissa ......................................................... 62<br />
Guimarães, J.A ........................................................ 222<br />
Guitián-Castrillón, José M. ................................ 77, 205<br />
Gupta, Divender ....................................................... 286<br />
Gurr, Geoff M. ................................................. 111, 264<br />
Gutierrez, J. Manuel ..................................................99<br />
Gutiérrez-Ruelas, José M. ........................................ 241<br />
Guzmán-Díaz, Miguel Á. .......................................... 257<br />
H<br />
Hai-Tung Feng ......................................................... 288<br />
Hamacek, Edward ......................................................55<br />
Hammouti, Nasera ................................................... 187<br />
Handler, Alfred M. ................................ 65, 67, 189, 191<br />
Hanna, Rachid ..................................... 96, 112, 123, 287<br />
Haq, Ihsan ul ...................................................... 87, 219<br />
Harris, A.R. .............................................................. 111<br />
Hattingh, Vaughan .....................................................76<br />
Heath, Robert R. ....................................... 284, 295, 302<br />
Hemdan, Heitham .................................................... 190<br />
Hendrichs, Jorge ................................... 83, 87, 144, 245<br />
Hennessey, Michael K. ...............................................75<br />
Hernán Camacho, V. ................................................ 156<br />
Hernández, Doris...................................................... 240<br />
Hernández, Emilio .............................................. 82, 233<br />
Hernández, M. Refugio ................................ 84, 89, 224<br />
Hernandez, Sofia ...................................................... 192<br />
Hernández-Crespo, Pedro ......................... 184, 185, 276<br />
Hernández-Ortiz, Vicente ..........................................61<br />
Ho, Kun-Yaw............................................................ 121<br />
Hoffmeister, Thomas S. ............................................ 262<br />
Hoskovec, Michal ..................................................... 173<br />
Hsu, Ju-Chun ................................................... 121, 288<br />
Huang, Yu-Bin ................................................. 101, 242<br />
Hunasaghatta, Praveen M. ....................................... 243<br />
Hurtrel, Béatrice ...................................................... 131<br />
I<br />
Ibañez-Palacios, Jorge ........................................ 85, 188<br />
Imoulan, Abdessamad .............................................. 258<br />
Ingram, Donald ........................................................ 135<br />
Islam, Amirul .............................................................91<br />
J<br />
Jacas, Josep A. .................................................. 226, 291<br />
Jackson, Kevin ...........................................................55<br />
Jacob, Arthur ........................................................... 250<br />
Jacquard, Cathy .................................. 49, 131, 150, 186<br />
Jamroenma, Kriengkai............................................. 171<br />
Jang, Eric ........................................ 41, 68, 95, 289, 309<br />
Jerez, Carlos ............................................................. 143<br />
Jessup, Andrew J. ..................... 60, 83, 91, 111, 219, 264<br />
329
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Jiang, Joe-Air ........................................................... 121<br />
Jiang, L. ................................................................... 225<br />
Jiji, T. ................................................................ 250, 290<br />
Joachim-Bravo, Iara S. ............................................ 232<br />
Johnson, Jerome ...................................................... 221<br />
Jordaens, Kurt ......................................................... 177<br />
Juan-Blasco, María ........................................... 226, 291<br />
Jumroenma, Kriengkrai ............................................. 259<br />
Júnior, Cyro R. C..................................................... 173<br />
Justo, Adrian D. ....................................................... 214<br />
K<br />
Kahrarian, Morteza ................................................. 161<br />
Kakani, Evdoxia ...................................................... 197<br />
Kalinová, Blanka ..................................................... 173<br />
Kao, Ching-Hua ................................................ 101, 242<br />
Katepongpun, Wichit ............................................... 234<br />
Katsoyannos, Byron ................................................. 125<br />
Kendra, Paul E.......................................... 284, 295, 302<br />
Khamis, Fathiya ......................................................... 42<br />
Khayrattee, Fazilla B. .............................................. 245<br />
Khlawi, Samira ........................................................ 292<br />
Khriman, Ashot ....................................................... 289<br />
Kittelmann, Sebastian ................................................ 65<br />
Kotze, Charl ............................................................. 128<br />
Koukidou, Martha ..................................................... 66<br />
Kouloussis, Nikos ..................................................... 125<br />
Kriticos, Darren J ...................................................... 76<br />
L<br />
Lacasa, Alfredo ................................................. 206, 207<br />
Lanzavecchia, Silvia ................................................. 254<br />
Lanzavecchia, Silvia B. ............................................ 255<br />
Laskar, Nripendra ................................................... 281<br />
Layme, Javier M. ....................................................... 75<br />
Leal, Rubén ................................................................ 99<br />
Lens, Luc.............................................................69, 155<br />
Lereculeur, Anabel .................................................. 131<br />
Li, Bai-Ni.................................................................. 215<br />
Liedo, Pablo ................................. 83, 113, 135, 144, 269<br />
Liendo, M. Clara ........................................... 48, 86, 152<br />
Lin, Tzu-Shiang ....................................................... 121<br />
Lindhout, K. ............................................................. 111<br />
Liquido, Nicanor J. .................................................... 75<br />
Liu, Kevin J.............................................................. 121<br />
Liu, Lu-Yan ............................................................... 59<br />
Lomeli Flores, J. Refugio ........................................... 98<br />
Lopes de Castro, Jairo ............................................. 163<br />
Lopes, D.J.H............................................................. 208<br />
Lopes, Fabiana ......................................................... 229<br />
López, Karim ........................................................... 227<br />
López, María M. ...................................................... 211<br />
López-Martínez, Victor ........................................... 256<br />
Lu, Kuang-Hui ........................................................... 59<br />
Lucas, Alfonso .................................................. 206, 207<br />
Lysandrou, Michael .......................................... 127, 298<br />
M<br />
M’saad Guerfali, Meriem ........................................ 190<br />
Macedo, N. ............................................................... 208<br />
Machota Jr., Ruben .......................................... 279, 280<br />
Maerere, Amon Petro .............................................. 300<br />
Mafra-Neto, Agenor........................... 127, 279, 280, 296<br />
Malacrida, Anna ........................ 42, 65, 67, 96, 196, 323<br />
Malavasi, Aldo ......................................................... 244<br />
Mangan, Robert L. .................................................... 78<br />
Mankin, Richard W. .................................................. 54<br />
Manrakhan, Aruna ........................................... 128, 213<br />
Mararuai, Amanda .................................................. 209<br />
Marco-Noales, Ester ................................................ 211<br />
Marcotegui, Aránzazu ............................................. 305<br />
Marín, Cándido ....................................................... 124<br />
Martí, Salvador ........................................................ 129<br />
Martínez, Marta ...................................................... 226<br />
Martínez-Ferrer, María T. ....................... 149, 160, 297<br />
Martinez-Sañudo, Isabel .......................................... 195<br />
Martínez-Utrillas, Miguel Ángel .............................. 226<br />
Martins, David S. ..................................................... 260<br />
Marusic, Ivana ......................................................... 239<br />
Marzal, Carmen....................................................... 160<br />
Mastrangelo, Thiago ......................................... 244, 315<br />
Matallanas, Beatriz .................................................. 192<br />
Matebe, Yolanda ...................................................... 213<br />
Mathiopoulos, Kostas ....................................... 190, 197<br />
Matioli, André L. ..................................................... 170<br />
Mau, Ronald .............................................................. 95<br />
Mavragani-Tsipidou, Penelope ................................ 197<br />
Mavrotas, Costas ...................................... 127, 130, 298<br />
Mazih, Ahmed.......................................................... 187<br />
Mazzon, Luca........................................................... 195<br />
Mbaye, Ndiaye ......................................................... 299<br />
McInnis, Donald ........................................................ 90<br />
Meats, Alan ................................................................ 71<br />
Medeiros, Aida ......................................................... 293<br />
Medina Hernández, Rosa E. .................................... 256<br />
Medina, Pilar ........................................................... 143<br />
Mediouni-Ben Jemâa, Jouda ............................ 261, 277<br />
Mejia, Carlos Gonzalo ............................................... 53<br />
Merayo, Eduardo .............................................. 210, 214<br />
Merkel, Katharina ................................................... 262<br />
Messoudi, Soumaya ................................................. 261<br />
Mexia, A. ........................................................... 208, 282<br />
Meza, J. Salvador........................................ 85, 189, 228<br />
Midgarden, David .................................................... 284<br />
Milla, Fabián.............................................................. 86<br />
Milla, Fabián H. ................................... 48, 152, 236, 252
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
Mirak, Tohid N .......................................................... 47<br />
Mohamed, Samira A. ............................................... 112<br />
Mohammadi, M. ...................................................... 153<br />
Mohammed Ali, Hussain ......................................... 292<br />
Mohammed Z. Khalaf ............................................. 294<br />
Mohammed, Samira A............................................. 251<br />
Mongabure, Alejandro P. .......................................... 97<br />
Montes, Sônia M. N. M. ..................... 157, 158, 170, 212<br />
Montgomery, Wayne S. .................................... 295, 302<br />
Montoya, Diego .......................................................... 53<br />
Montoya, Pablo .................................... 82, 113, 257, 269<br />
Monzó, César ........................................................... 109<br />
Moraiti, Cleopatra A. .............................................. 159<br />
Morales-Valles, Pedro ............................................... 61<br />
Moreira, M.A.B ....................................................... 222<br />
Morelli, Renata ................................................... 90, 229<br />
Morgan, Sian A.......................................................... 66<br />
Moya, Pilar ....................................................... 107, 266<br />
Moya-Sanz, Pilar ..................................................... 285<br />
Mulkay, Tania ......................................................... 240<br />
Mumford, John M. ................................... 102, 208, 290<br />
Muniyappa, Nandeesh ............................................. 243<br />
Mwatawala, Maulid .................................... 42, 155, 300<br />
N<br />
Nascimento, Antônio S............................................. 232<br />
Naseema, A .............................................................. 290<br />
Naseema, T .............................................................. 250<br />
Navack, Kelly I. ....................................................... 166<br />
Navarro, Ismael ....................................................... 120<br />
Navarro-Campos, Cristina ...................................... 160<br />
Navarro-Fuertes, Ismael .......................................... 285<br />
Navarro-Llopis, Vicente .................... 126, 274, 285, 301<br />
Nave, Anabela .......................................................... 143<br />
Nicol. H. I. ................................................................ 225<br />
Nikpay, Amin........................................................... 161<br />
Niogret, Jerome ....................................................... 302<br />
Nirmala, Xavier ....................................................... 191<br />
Nisha, V.G ............................................................... 290<br />
Noikeaing, Sirilux .................................................... 303<br />
Nolazco, Norma ................................................ 179, 227<br />
Novelo-Rincón, Luisa F. .......................................... 269<br />
Nyamukondiwa, Casper ............................................ 51<br />
O<br />
Ochando, María Dolores .................................... 72, 192<br />
Ogaugwu, Christian E. ............................................ 193<br />
Olaniyi, Mercy ......................................................... 318<br />
Oliveira, G. B. .......................................................... 232<br />
Oliveira, Luísa ......................................................... 293<br />
Orankanok, Somchai ................................ 234, 235, 304<br />
Orankanok, Watchreeporn . 88, 223, 230, 234, 235, 246,<br />
303, 304<br />
Ordax, Mónica ......................................................... 211<br />
Oropeza, Azucena .............................................. 83, 269<br />
Orozco-Dávila, Dina ............................................. 84, 89<br />
Ortego, Félix .............................................. 184, 185, 276<br />
Ortiz-Urquiza, Almudena ........................................ 105<br />
Osorio, Armando ...................................................... 178<br />
Ouagoussounon, Issa ........................................ 110, 319<br />
Ouédraogo, S. Nafiba ............................................... 162<br />
Oufdou, Khalid ......................................................... 260<br />
P<br />
Palència, Jaume ........................................................ 273<br />
Papadopoulos, Nikos T. .............. 125, 136, 137, 159, 183<br />
Papanastasiou, Stella ................................................ 137<br />
Papeschi, Alba G. ..................................................... 254<br />
Paranhos, Beatriz J .....................................90, 229, 232<br />
Paranhos, Lucas G .....................................................90<br />
Paredes, Daniel ......................................................... 143<br />
Paroonagian, Doris ........................................... 127, 298<br />
Parreño, María A. ......................................................48<br />
Pascual, Felipe .......................................................... 143<br />
Pascual, Susana ................................................ 143, 305<br />
Paula, Laís Ívina Silva de ......................................... 163<br />
Paumier, Adrian ....................................................... 240<br />
Payet, Christine ..........................................................49<br />
Peña, Leandro .......................................................... 312<br />
Peñarrubia-María, Esther ................................ 306, 308<br />
Peralta, Patricia A. ............................................. 86, 236<br />
Pereira, André F. .............................................. 201, 202<br />
Pereira, José A. ................................................. 143, 282<br />
Pereira, Júlia D. B. ........................................... 168, 169<br />
Perera, Nathalie ................................................ 185, 276<br />
Pérez-Hinarejos, Marta ............................................ 253<br />
Pérez-Lachaud, Gabriela ......................................... 113<br />
Pérez-Panadés, Jordi ................................................ 231<br />
Pérez-Staples, Diana ................................................. 228<br />
Permalloo, Shradhanand .......................................... 245<br />
Pimentel, R. .............................................................. 208<br />
Pinkaew, Supaap .........................................88, 223, 230<br />
Piquer-Salcedo, Jaime E. .......................................... 211<br />
Piyarath, Sudsa-nguan ............................................. 303<br />
Plá, Ignacio ............................................................... 226<br />
Planes, Laura............................................................ 291<br />
Plodkornburee, Wipada ................................... 171, 259<br />
Popovic, Luka ........................................................... 239<br />
Porcel, Mario ............................................................ 143<br />
Pratt, C.F. ................................................................. 111<br />
Primo, Jaime ...................... 107, 126, 266, 274, 285, 301<br />
Pujade-Villar, Juli .................................................... 124<br />
Putruele, Graciela .................................................... 307<br />
331
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Q<br />
Quesada-Moraga, Enrique ............................... 105, 263<br />
Quilici, Serge ..........49, 131, 148, 150, 151, 186, 306, 308<br />
Quintanilla, Luis ............................................... 147, 154<br />
Quinto, Vicente ................................................. 206, 207<br />
Quiroga, Diego ......................................................... 100<br />
R<br />
Rabelo, Lílian R. S. ........................................... 201, 202<br />
Raga, Adalton ............................ 157, 158, 163, 170, 212<br />
Raharimalala, Soanjanahary ................................... 310<br />
Raies, Aly ................................................................. 190<br />
Ramadan, Mohsen M. .............................................. 115<br />
Rambhunjun, Mahen ............................................... 245<br />
Ramírez, Wilda ........................................................ 100<br />
Ramsey, Amanda .............................................. 289, 309<br />
Raoelijaona, Adrien R. ............................................ 310<br />
Raoelijaona, Jeannette C. Y. .................................... 310<br />
Rasamizafy, Lerry A. ............................................... 310<br />
Rasgado, Milton ......................................................... 82<br />
Rasool, Bilal ........................................................47, 164<br />
Ratovonomenjanahary, Zelin T. .............................. 310<br />
Rattanapun, Wigunda.........................................50, 165<br />
Rei, Fernando .......................................................... 143<br />
Rendon, José Raúl ..................................................... 53<br />
Restrepo-Ortiz, Claudia X. ...................................... 124<br />
Reuveny, Haim......................................................... 311<br />
Reyes, Jesus.............................................................. 144<br />
Reynolds, Olivia L............................................. 111, 264<br />
Rial, Esteban J. .......................................................... 97<br />
Richards, Stephen ...................................................... 67<br />
Riegler, Markus ..................................................47, 164<br />
Rios, Anderli D. F. ............................................ 201, 202<br />
Rivera, Pedro ........................................................... 233<br />
Robinson, Alan....................................................87, 144<br />
Rodrigo, M.Jesús ..................................................... 312<br />
Rodríguez Leyva, Esteban ......................................... 98<br />
Rodríguez, Ana ........................................................ 312<br />
Rodriguez, Jorge L. ................................................. 240<br />
Romero, Moises.......................................................... 82<br />
Ronchi-Teles, Beatriz ................................................ 180<br />
Ros, J. Pedro ...................................... 143, 278, 313, 314<br />
Rosillo, Mario .......................................................... 210<br />
Rouland-Lefèvre Corinne ........................................ 162<br />
Roura, Lídia ............................................................. 273<br />
Rousse, Pascal .......................................................... 131<br />
Rovelo M., Rosa E. ................................................... 224<br />
Ruano, Francisca ..................................................... 143<br />
Ruiz, Cynthia ........................................................... 100<br />
Ruiz, J. Manuel ........................................................ 314<br />
Ruiz, Joaquín ........................................................... 314<br />
Ruiz, Lía................................................................... 113<br />
Ruíz, Manuel ............................................................ 143<br />
Ruiz-Arce, Raul ....................................................... 194<br />
Rull, Juan................................................................... 62<br />
Rwegasira, Gration .................................................. 300<br />
S<br />
Sabater Alepuz, Juan B. .......................................... 283<br />
Sabater-Muñoz, Beatriz ..... 109, 211, 226, 260, 265, 291,<br />
324<br />
Salcedo Baca, Diznarda ............................................. 98<br />
Salin, Daniel ............................................................. 203<br />
Salvador, Miguel ...................................................... 233<br />
San Andrés, Victoria ........... 107, 231, 265, 266, 312, 324<br />
Sánchez-Ramos, Ismael .................................... 143, 305<br />
Sanchis, Juan .................................................... 126, 301<br />
Santiago-Álvarez, Cándido ............................... 105, 263<br />
Santiago-Martínez, Guillermo ................................. 241<br />
Santos, Mirian S. ..................................................... 166<br />
Santos, Deise C. C. ................................................... 232<br />
Santos, Tatiana R. O. ............................................... 232<br />
Sarika, Mohan ......................................................... 290<br />
Savio, Claudia .......................................................... 195<br />
Scherer, Steven .......................................................... 67<br />
Schetelig, Marc F. ................................. 65, 67, 191, 193<br />
Schmitt, Chloé................................................... 306, 308<br />
Schuler, Hannes .................................................. 47, 164<br />
Schütze, Mark K. ................................................. 60, 91<br />
Scolari, Francesca ....................................... 65, 196, 323<br />
Segade, Gonzalo ....................................................... 307<br />
Segura, Diego F. ..................... 48, 86, 152, 236, 252, 255<br />
Segura, M. Dolores .................................................... 72<br />
Seitz, Alfred ............................................................. 187<br />
Selivon, Denise ........................................................... 61<br />
Sellini, Lotfi ............................................................. 190<br />
Seris, Elena ............................................... 143, 305, 313<br />
Sermann, Helga ....................................................... 249<br />
Setyawati, W. ........................................................... 172<br />
Shanmugam, Vijaysegaran ...................................... 243<br />
Sheela, M.S. ............................................................. 250<br />
Shimada, Takehiko .................................................. 312<br />
Siciliano, Paolo .................................................. 196, 323<br />
Siderhurst, Matthew ................................................ 289<br />
Silva Neto, Alberto M. ............................................. 232<br />
Silva, Janisete G. ............................................... 166, 167<br />
Silva, Márcio A. ....................................................... 315<br />
Silva, Mariangela V. L. ............................................ 232<br />
Silva, N. M. O. .......................................................... 167<br />
Silva, Ricardo A. ............................................... 168, 169<br />
Simiand, Christophe ................................. 131, 150, 186<br />
Sinzogan, Antonio ............................................. 110, 319<br />
Sittipitak, Wanlapa .................................................. 303<br />
Sivinski, John .................................................... 114, 268<br />
Skouri, Wafa ............................................................ 260<br />
Smaili, Charif........................................................... 316<br />
Solís E., Eduardo A. ................................................. 224
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
Sookar, Preeaduth ................................................... 245<br />
Souleymane, Lessueur ............................................. 299<br />
Srikachar, Sunyanee ......................................... 171, 259<br />
Stauffer, Christian .............................................. 47, 164<br />
Steck, Gary J. ........................................................... 180<br />
Stoltman, Lyndsie 1 ................................................... 296<br />
Stones, Wilna ........................................................... 213<br />
Stuhl, Charles .......................................................... 268<br />
Sudhi-Aromna, Sarute ............................................... 259<br />
Suja, G. .................................................................... 290<br />
Sunandita................................................................. 286<br />
Sundaralingam, S. ................................................... 225<br />
Susanto, A. ............................................................... 172<br />
Sutherland, Ian ........................................................ 204<br />
Sved, John.................................................................. 71<br />
Syamsudin, Tati Suryati .......................................... 172<br />
T<br />
Tadeo, Eduardo ......................................................... 62<br />
Tarshis Moreno, Aleena ............................................ 78<br />
Tavares, Laura ........................................................ 293<br />
Teal, Peter..................................................... 86, 87, 268<br />
Tejeda, Marco T. ..................................................... 228<br />
Telavanich, Thitipong .............................................. 246<br />
Terblanche, Jonh S. ................................................... 51<br />
Terrazas González, Gerardo H.................................. 98<br />
Tescari, Enzo ........................................................... 298<br />
Thaochan, Narit......................................................... 56<br />
Thomas, John .......................................................... 318<br />
Thumdee, Patama .................................................... 234<br />
Toledo, Jorge ...................................................... 83, 269<br />
Tormos, José ............................................................ 253<br />
Torné, Maria .................................................... 130, 298<br />
Torrent, J. ................................................................ 263<br />
Torres, Laura .......................................................... 143<br />
Tortosa, A. ............................................................... 278<br />
Trelluyer, Mickael ................................................... 308<br />
Tsai, Wei-Huang ........................................................ 59<br />
Tsoumani, Konstantina ........................................... 197<br />
U<br />
Uggla, Madeleine ..................................................... 317<br />
Umeh,Vincent .......................................................... 318<br />
Urbaneja, Alberto .............................. 109, 226, 231, 291<br />
Utgés, M. Eugenia ...................................................... 86<br />
V<br />
Vacas, Sandra ........................................... 126, 274, 301<br />
Van Mele, Paul ................................................. 110, 319<br />
Vangsilabutr, Wipada ................................................ 259<br />
Vaníčková, Lucie ...................................................... 173<br />
Vargas, Roger ...................................... 95, 127, 289, 296<br />
Vayssières, Jean-François ................... 96, 110, 162, 319<br />
Vázquez López, Margarito ....................................... 256<br />
Velapatiño, Jorge F. ...................................................75<br />
Veloso, Valquíria da R. S. ................................ 201, 202<br />
Vendramim, José D. ................................................. 315<br />
Venter, Jan Hendrik ................................................. 213<br />
Ventura, L.B. ............................................................ 208<br />
Vera, M. Teresa ....................................... 48, 52, 86, 152<br />
Verghese, Abraham .......................................... 102, 290<br />
Vermeulen, Nathan .................................................. 221<br />
Vidal-Quist, J. Cristian ............................................ 106<br />
Vilajeliu, Marià ........................................................ 306<br />
Villalba, Daniel Nestor ............................................. 214<br />
Villaseñor, Antonio ............................................... 82, 99<br />
Viñuela, Elisa............................................................ 143<br />
Virgilio, Massimiliano ............................. 42, 69, 70, 177<br />
Virginio, Jair F. ........................................................ 232<br />
Viscarret, Mariana M. ...................................... 252, 255<br />
W<br />
Wadjinny, Jamila ..................................................... 316<br />
Walder, Julio M. M. ........................................... 90, 232<br />
Walder, Julio M.M. .................................................. 244<br />
Wang, Bo .................................................................. 320<br />
Watechakul, Narumon ............................................. 235<br />
Wattier, Christopher ................................................ 131<br />
Willink, Eduardo ........................................................52<br />
Wimmer, Ernst A. .............................................. 65, 193<br />
Wong, Lyle .................................................................95<br />
Wornoayporn, Viwat .......................................... 91, 219<br />
X<br />
Xi, Zhiyong ............................................................... 323<br />
Y<br />
Yang, En-Cheng ....................................................... 121<br />
Yokoyama, Victoria Y. ............................................. 108<br />
Z<br />
Zacarías, Lorenzo ..................................................... 312<br />
Zavala, J. Luis ............................................................99<br />
Zeni, Diego................................................................ 296<br />
Zepeda-Cisneros, Cristina S. ............... 85, 174, 188, 189<br />
Zhang, Run-Jie ......................................................... 215<br />
Zorman, M. .............................................................. 208<br />
Zou, Sige ................................................................... 135<br />
333
8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
Zucchi, Roberto A. ............................................ 168, 169
NOTES<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
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NOTES<br />
8 th International Symposium on Fruit Flies of Economic Importance. Valencia (Spain) 26 th September to 1 st October<br />
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8 th International Symposium on Fruit Flies of Economic Importance. Valencia 2010.<br />
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