contribution to session IV

Test methodolog

Aupinel, Pierrick

Fortini, Dominique

Michaud, Bruno

English

JKI Open Journal Systems (Julius Kühn-Institut)

Hazards of pesticides to bees – 10th International Symposium of the ICP-Bee Protection Group 102 Julius-Kühn-Archiv 423, 2009 Acknowledgements The authors would like to thank: Markus Barth (Biochem agrar, Germany), Stephan Schmitzer (IBACON, Germany), Silvio Scazzari (IBACON, Germany), Jens Pistorius (JKI, Germany, formerly eurofins-GAB, Germany), Hans Ulrich Rexer (eurofins-GAB, Germany), Marie-Pierre Chauzat (AFSSA, France) who ran the preliminary experiments but for different reasons could not run the ring test. References 1 Commission Directive 96/12/EC, Amendment to Council Directive 91/414/EEC. Official Journal of the European Communities No. L 65: 20 – 37 (1996). 2 Oomen PA, De Ruijter A and Van der Steen J, Method for honeybee brood feeding tests with insect growth – regulating insecticides. Bull OEPP / EPPO Bull 22: 613-616 (1992). 3 Schur A, Tornier I, Brasse D, Mühlen W, Von der Ohe W, Wallner K and Wehling M, Honey bee brood ring-test in 2002: method fort the assessment of side effects of plant protection products on the honey bee brood under semi-field conditions. Bull Insectol 56(1): 91-96 (2003). 4 OECD (2007): Guidance document on the honeybee (Apis mellifera L.) brood test under semi-field conditions. OECD Environment, Health and Safety Publications, Series on Testing and Assessment No. 75, ENV/JM/MONO (2007) 22 5 Aupinel P, Fortini D, Dufour H, Tasei JN, Michaud B, Odoux JF and Pham Delègue MH, Improvement of artificial feeding in a standard in vitro method for rearing Apis mellifera larvae. Bull Insectol 58 (2): 107-111 (2005). 6 Aupinel P, Fortini D, Dufour H, Michaud B, Marolleau F, Tasei JN and Odoux JF, Toxicity of dimethoate and fenoxycarb to honey bee brood (Apis mellifera), using a new in vitro standardized feeding method. Pest Manag Sci 63: 1090-1094 (2007). 7 Aupinel P, Medrzycki P, Fortini D, Dufour H, Michaud B, Tasei JN and Odoux JF, A new larval in vitro rearing method to test effects of pesticides on honey bee brood. Redia XC: 87-90 (2007). 8 Gough HJ, McIndoe EC and Lewis GB, The use of dimethoate as a reference compound in laboratory acute toxicity tests on honey bees (Apis mellifera L.) 1981-1982. J. Apic. Res. 33(2): 119-125 (1994).  Comparison of two methods to assess effects of insecticides on hypopharyngeal gland development of honey bee Dominique Fortini, Bruno Michaud, Pierrick Aupinel Unité expérimentale d’entomologie, INRA, Le Magneraud, BP 52, 17700 Surgères, France Abstract Hypopharyngeal glands (HPG) are the main organs responsible of royal jelly secretion. The size of the HPG is age2, 4 and food protein5, 7 dependent, and correlated to the amount of secretion2, and the weight of the head5. Their development can be assessed with a microscope by measuring the acini diameter after dissection.. This very useful method1, 3, 5, 6 has some inconveniences: it requires dexterity to extract the gland, and the diameter of the acini is difficult to measure because of its pear shape. In order to assess the HPG development, total protein of the gland can be measured with the Bradford method7, 8, 9, but this also requires to extract it from the head. The development of the HPG may be also affected by substances known for their insecticide effects like soybean tripsin inhibitor 8, 9. The objective of this work is to compare two methods for assessing the effects of insecticides on HGP development. The first one consists in measuring the acini diameter, and the second one in measuring the total protein of the head. The measurements are made on bee nurses intoxicated during 10 days with sublethal doses of dimethoate. Hazards of pesticides to bees – 10th International Symposium of the ICP-Bee Protection Group Julius-Kühn-Archiv 423, 2009 103 References 1 Babendreier D., Kalberer N. K., Romeis J. , Fluri P., Mulligan E., Bigler F. (2005). Influence of Bt-transgenic pollen, Bt-toxin and protease inhibitor (SBTI) ingestion on development of the hypopharyngeal glands in honeybees. Apidology 36: 584-594. 2 Deseyn J., Billen J. (2005) Age-dependent morphology and ultrastructure of the hypopharyngeal gland of Apis mellifera workers (Hymenoptera, Apidae). Apidology 36: 49-57. 3 Gupta P.R., Chandel R. S. (1994) Effects of diflubenzuron and penfluron on workers of Apis cerana and Apis mellifera L. Apidology 26: 3-10. 4 Huang Z. Y., Otis G. W. (1989) Factors determining hypopharyngeal gland activity of worker honey bees (Apis mellifera L.). Insectes sociaux 36(4): 264-276. 5 Hrassnigg N., Crailsheim K. (1998) Adaptation of hypopharyngeal gland development to the brood status of honeybee (Apis mellifera L.) colonies. Journal of insect Physiology 44: 929-939. 6 Malone L. A., Todd H. J., Burgess E. P. J., Chriteller J. T. (2004) Development of hypopharyngeal glands in adult honey bees fed with Bt toxin, a biotin-binding protein and protease inhibitor. Apidology, 35: 655-664. 7 Pernal S. F., Currie R. W. (2000) Pollen quality of fresh and 1-year-old single pollen diets for worker honey bees (Apis mellifera L.). Apidology 31: 387-409. 8 Sagili R. R., Pankiw T., Zhu-salsman K. (2005) Effects of soybean inhibitor on hypopharyngeal gland protein content, total midgut protease activity and survival of the honey bee (Apis mellifera L.). Journal of insect Physiology 51: 953-957. 9 Sagili R. R., Pankiw T. (2007) effects of protein-constrained brood food on honey bee (Apis mellifera L.) pollen foraging and colony growth. Behavioral Ecology and Sociobiology 61:1471-1478.  V. Honey bee poisoning incidents and monitoring schemes Review of honeybee pesticide poisoning incidents in Europe – evaluation of the hazard quotient approach for risk assessment Helen M Thompson1*, David Thorbahn2 1 CSL, Sand Hutton, York, UK YO41 1LZ 2 Biologische Bundesanstalt für Land- und Forstwirtschaft, Untersuchungsstelle für Bienenvergiftungen, Messeweg 11-12 , 38104 Braunschweig, Germany *email: h.thompson@csl.gov.uk; Tel: 44 1904 462515; fax 44 1904 462515 Abstract Background: Honeybee risk assessment is required in Europe for all pesticides where bees may be exposed. This is well established for sprayed products where the hazard quotient (HQ), calculated by dividing the application rate of the sprayed product active ingredient by the LD50, is less than 50 the product is considered safe to bees (unless it is an IGR).  In the UK, Germany and the Netherlands post-registration monitoring schemes on the poisoning of honeybee by pesticides collate data on honeybee incidents.  Results: The incident schemes have been invaluable in identifying agronomic practices resulting in honeybee mortality and changes have been made to labelling to address such issues.  The decrease in the numbers of incidents reported supports the assertion that such schemes have positively contributed to the regulatory process and also provide confidence in the risk assessment approaches. Conclusion: This review of incidents in Europe over the last 25 years suggests that the HQ approach to risk assessment for honeybees offers an appropriate level of protection. Keywords: honeybee, pesticide, hazard quotient, risk assessment 

Comparison of two methods to assess effects of insecticides on hypopharyngeal gland development of honey bee

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Comparison of two methods to assess effects of insecticides on hypopharyngeal gland development of honey bee

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