Advertisement

Ecotoxicology

, Volume 21, Issue 8, pp 2214–2221 | Cite as

Does transgenic Cry1Ac + CpTI cotton pollen affect hypopharyngeal gland development and midgut proteolytic enzyme activity in the honey bee Apis mellifera L. (Hymenoptera, Apidae)?

  • Peng Han
  • Chang-Ying NiuEmail author
  • Antonio Biondi
  • Nicolas Desneux
Article

Abstract

The transgenic Cry1Ac (Bt toxin) + CpTI (Cowpea Trypsin Inhibitor) cotton cultivar CCRI41 is increasingly used in China and potential side effects on the honey bee Apis mellifera L. have been documented recently. Two studies have assessed potential lethal and sublethal effects in young bees fed with CCRI41 cotton pollen but no effect was observed on learning capacities, although lower feeding activity in exposed honey bees was noted (antifeedant effect). The present study aimed at providing further insights into potential side effects of CCRI41 cotton on honey bees. Emerging honey bees were exposed to different pollen diets using no-choice feeding protocols (chronic exposure) in controlled laboratory conditions and we aimed at documenting potential mechanisms underneath the CCRI41 antifeedant effect previously reported. Activity of midgut proteolytic enzyme of young adult honey bees fed on CCRI41 cotton pollen were not significantly affected, i.e. previously observed antifeedant effect was not linked to disturbed activity of the proteolytic enzymes in bees’ midgut. Hypopharyngeal gland development was assessed by quantifying total extractable proteins from the glands. Results suggested that CCRI41 cotton pollen carries no risk to hypopharyngeal gland development of young adult honey bees. In the two bioassays, honey bees exposed to 1 % soybean trypsin inhibitor were used as positive controls for both midgut proteolytic enzymes and hypopharyngeal gland proteins quantification, and bees exposed to 48 ppb (part per billion) (i.e. 48 ng g−1) imidacloprid were used as controls for exposure to a sublethal concentration of toxic product. The results show that the previously reported antifeedant effect of CCRI41 cotton pollen on honey bees is not linked to effects on their midgut proteolytic enzymes or on the development of their hypopharyngeal glands. The results of the study are discussed in the framework of risk assessment of transgenic crops on honey bees.

Keywords

Hypopharyngeal gland Midgut enzymes Risk assessment Sublethal effects Imidacloprid Transgenic cotton pollen 

Notes

Acknowledgments

We are grateful to Xiaoxia Dong, Shifeng Yu and Haixia Yang for technical help for the dissection works on honey bees during the study, and to JB Yang for technical assistance for honey bee rearing. This work was supported by the National Natural Science Foundation of China (Grant No. 31071690). The authors declare that they have no conflict of interest.

Supplementary material

10646_2012_976_MOESM1_ESM.doc (30 kb)
Supplementary material 1 (DOC 29 kb)

References

  1. Abbott WS (1925) A method for computing the effectiveness of an insecticide. J Econ Entomol 18:265–267Google Scholar
  2. Arno J, Gabarra R (2011) Side effects of selected insecticides on the Tuta absoluta (Lepidoptera: Gelechiidae) predators Macrolophus pygmaeus and Nesidiocoris tenuis (Hemiptera: Miridae). J Pest Sci 84:513–520CrossRefGoogle Scholar
  3. Babendreier D, Kalberer NM, 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. Apidologie 36:585–594CrossRefGoogle Scholar
  4. Babendreier D, Joller D, Romeis J, Bigler F, Widmer F (2007) Bacterial community structures in honeybee intestines and their response to two insecticidal proteins. FEMS Microbiol Ecol 59:600–610CrossRefGoogle Scholar
  5. Bakhsh A, Rao AQ, Shahid AA, Husnain T, Riazuddin S (2010) Camv 35S is a developmental promoter being temporal and spatial in expression pattern of insecticidal genes (Cry1ac & Cry2a) in cotton. Aust J Basic Appl Sci 4:37–44Google Scholar
  6. Biondi A, Desneux N, Siscaro G, Zappalà L (2012) Using organiccertified rather than synthetic pesticides may not be safer for biological control agents: selectivity and side effects of 14 pesticides on the predator Orius laevigatus. Chemosphere 87:803–812CrossRefGoogle Scholar
  7. Bøhn T, Traavik T, Primicerio R (2010) Demographic responses of Daphnia magna fed transgenic Bt-maize. Ecotoxicology 19:419–430CrossRefGoogle Scholar
  8. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  9. Brittain C, Bommarco R, Vighi M, Barmaz S, Settele J, Potts SG (2010) The impact of an insecticide on insect flower visitation and pollination in an agricultural landscape. Agr For Entomol 12:259–266Google Scholar
  10. Carrière Y, Ellers-Kirk C, Cattaneo MG, Yafuso CM, Antilla L, Huang CY, Rahman M, Orr BJ, Marsh SE (2009) Landscape effects of transgenic cotton on non-target ants and beetles. Basic Appl Ecol 10:597–606CrossRefGoogle Scholar
  11. Chen LZ, Cui JJ, Ma WH, Niu CY, Lei CL (2011) Pollen from Cry1Ac/CpTI-transgenic cotton does not affect the pollinating beetle Haptoncus luteolus. J Pest Sci 84:9–14CrossRefGoogle Scholar
  12. Clive J (2009) Global status of commercialized biotech/GM crops: 2009, The first fourteen years, 1996 to 2009. ISAAA brief no 41. ISAAA, Ithaca, NYGoogle Scholar
  13. Cui JJ (2003) Effects and mechanisms of the transgenic Cry1Ac plus CpTI (cowpea trypsin inhibitor) cotton on insect communities. Dissertation, Chinese Academy of Agricultural SciencesGoogle Scholar
  14. Dai PL, Zhou W, Zhang J, Cui HJ, Wang Q, Jiang WY, Sun JH, Wu YY, Zhou T (2012a) Field assessment of Bt cry1Ah corn pollen on the survival, development and behavior of Apis mellifera ligustica. Ecotoxicol Environ Safe 79:232–237CrossRefGoogle Scholar
  15. Dai PL, Zhou W, Zhang J, Jiang WY, Wang Q, Cui HJ, Sun JH, Wu YY, Zhou T (2012b) The effects of Bt Cry1Ah toxin on worker honeybees (Apis mellifera ligustica and Apis cerana cerana). Apidologie 43:384–391CrossRefGoogle Scholar
  16. Decourtye A, Mader E, Desneux N (2010) Landscape scale enhancement of floral resources for honey bees in agro-ecosystems. Apidologie 41:264–277CrossRefGoogle Scholar
  17. Decourtye A, Pham-Delègue MH (2002) The proboscis extension response: assessing the sublethal effects of pesticides on the honey bee. In: Devillers J, Pham-Delègue MH (eds) Honey Bees: Estimating the Environmental Impact of Chemicals. Taylor & Francis, London/New York, pp 67–81Google Scholar
  18. Desneux N, Bernal JS (2010) Genetically modified crops deserve greater ecotoxicological scrutiny. Ecotoxicology 19:1642–1644CrossRefGoogle Scholar
  19. Desneux N, Decourtye A, Delpuech JM (2007) The sublethal effects of pesticides on beneficial arthropods. Annu Rev Entomol 52:81–106CrossRefGoogle Scholar
  20. Desneux N, Ramirez-Romero R, Bokonon-Ganta AH, Bernal JS (2010) Attraction of the parasitoid Cotesia marginiventris to host frass is affected by transgenic maize. Ecotoxicology 19:1183–1192CrossRefGoogle Scholar
  21. Fluri P, Lüscher M, Wille H, Gerig L (1982) Changes in weight of the pharyngeal gland and haemolymph titres of juvenile hormone, protein and vitellogenin in worker honeybees. J Insect Physiol 28:61–68CrossRefGoogle Scholar
  22. Gassmann AJ, Carriere Y, Tabashnik BE (2009) Fitness costs of insect resistance to Bacillus thuringiensis. Annu Rev Entomol 54:147–163CrossRefGoogle Scholar
  23. Giurfa M (2003) Cognitive neuroethology: dissecting non-elemental learning in a honeybee brain. Curr Opin Neurobiol 13:726–735CrossRefGoogle Scholar
  24. Han P, Niu CY, Lei CL, Cui JJ, Desneux N (2010a) Quantification of toxins in a Cry1Ac + CpTI cotton cultivar and its potential effects on the honey bee Apis mellifera L. Ecotoxicology 19:1452–1459CrossRefGoogle Scholar
  25. Han P, Niu CY, Lei CL, Cui JJ, Desneux N (2010b) Use of an innovative T-tube maze assay and the proboscis extension response assay to assess sublethal effects of GM products and pesticides on learning capacity of the honey bee Apis mellifera L. Ecotoxicology 19:1612–1619CrossRefGoogle Scholar
  26. Haydak MH (1970) Honey bee nutrition. Annu Rev Entomol 15:143–156CrossRefGoogle Scholar
  27. He YX, Zhao J, Zheng Y, Zhan Z, Desneux N, Wu KM (2012) Lethal effect of imidacloprid on the coccinellid predator Serangium japonicum and sublethal effects on predator voracity and on functional response to the whitefly Bemisia tabaci. Ecotoxicology 21:1291–1300CrossRefGoogle Scholar
  28. Hendriksma HP, Härtel S, Steffan-Dewenter I (2011) Testing pollen of single and stacked insect-resistant Bt-maize on in vitro reared honey bee larvae. PLoS One 6:e28174. doi: 10.1371/journal.pone.0028174 CrossRefGoogle Scholar
  29. Hrassnigg N, Crailsheim K (1998) Adaptation of hypopharyngeal gland development to the brood status of honeybee (Apis mellifera L.) colonies. J Insect Physiol 44:929–939CrossRefGoogle Scholar
  30. Hyams DG (1993) CurveExpert Software Version 1.38: A curve fitting system for windows, MississippiGoogle Scholar
  31. Knecht D, Kaatz HH (1990) Patterns of larval food production by hypopharyngeal glands in adult worker honey bees. Apidologie 21:457–468CrossRefGoogle Scholar
  32. Laskowski M, Kato L (1980) Protein inhibitors of proteinases. Annu Rev Biochem 49:593–626CrossRefGoogle Scholar
  33. Li GP, Feng HQ, McNeil JN, Liu B, Chen PY, Qiu F (2011) Impacts of transgenic Bt cotton on a non-target pest, Apolygus lucorum (Meyer-Dür) (Hemiptera: miridae), in northern China. Crop Prot 30:1573–1578CrossRefGoogle Scholar
  34. Liu B, Shu C, Xue K, Zhou KX, Li XG, Liu DD, Zheng YP, Xu CR (2009) The oral toxicity of the transgenic Bt + CpTI cotton pollen to honey bees (Apis mellifera). Ecotox Environ Safe 72:1163–1169CrossRefGoogle Scholar
  35. Lu YH, Wu KM, Jiang YY, Guo YY, Desneux N (2012) Widespread adoption of Bt cotton and insecticide decrease promotes biocontrol services. Nature 487:362–365Google Scholar
  36. Malone LA, Pham-Delègue MH (2001) Effects of transgene products on honey bees (Apis mellifera) and bumblebees (Bombus sp.). Apidologie 32:287–304CrossRefGoogle Scholar
  37. Malone LA, Todd JH, Burgess EPJ, Christeller JT (2004) Development of hypopharyngeal glands in adult honey bees fed with a Bt toxin, a biotin-binding protein and a protease inhibitor. Apidologie 35:655–664CrossRefGoogle Scholar
  38. Michener CD (1974) The social behavior of the bees: a comparative study. Harvard University Press, CambridgeGoogle Scholar
  39. Moritz B, Crailsheim K (1987) Physiology of protein digestion in the midgut of the honeybee (Apis mellifera L.). J Insect Physiol 33:923–931CrossRefGoogle Scholar
  40. Patel NG, Haydak MH, Gochnauer TA (1960) Electrophoretic components of the proteins in honeybee larval food. Nature 186:633–634CrossRefGoogle Scholar
  41. Ramirez-Romero R, Josette C, Pham-Delègue MH (2005) Effects of Cry1Ab protoxin, deltamethrin and imidacloprid on the foraging activity and the learning performances of the honeybee Apis mellifera, a comparative approach. Apidologie 36:601–611CrossRefGoogle Scholar
  42. Ramirez-Romero R, Desneux N, Chaufaux J, Kaiser L (2008a) Bt-maize effects on biological parameters of the non-target aphid Sitobion avenae (Homoptera: Aphididae) and Cry1Ab toxin detection. Pestic Biochem Physiol 91:110–115CrossRefGoogle Scholar
  43. Ramirez-Romero R, Desneux N, Decourtye A, Chaffiol A, Pham-Delègue MH (2008b) Does Cry1Ab protein affect learning performance of the honey bee Apis mellifera L. (Hymenoptera, Apidae)? Ecotoxicol Environ Safe 70:327–333CrossRefGoogle Scholar
  44. Robinson GE (1992) Regulation of division of labor in insect societies. Annu Rev Entomol 37:637–665CrossRefGoogle Scholar
  45. Romeis J, Bartsch D, Bigler F et al (2008) Assessment of risk of insect-resistant transgenic crops to non-target arthropods. Nat Biotechnol 26:203–208CrossRefGoogle Scholar
  46. Rui YK, Wang BM, Li ZH, Duan LS, Tian XL, Zhai ZX, He ZP (2004) Development of an enzyme immunoassay for the determination of the cowpea trypsin inhibitor (CpTI) in transgenic crop. Sci Agric Sin 37:1575–1579Google Scholar
  47. Sagili RR, Pankiw T (2007) Effects of protein-constrained brood food on honey bee (Apis mellifera L.) pollen foraging and colony growth. Behav Ecol Sociobiol 61:1471–1478CrossRefGoogle Scholar
  48. Sagili RR, Pankiw T, Zhu-Salzman K (2005) Effects of soybean trypsin inhibitor on hypopharyngeal gland protein content, total midgut protease activity and survival of the honey bee (Apis mellifera L.). J Insect Physiol 51:953–995CrossRefGoogle Scholar
  49. SAS Institute (1999) SAS/Stat user’s guide, release 8 edn. SAS Institute, Cary, NCGoogle Scholar
  50. Siebert MW, Patterson TG, Gilles GJ, Nolting SP, Braxton LB, Leonard BR, Van Duyn JW, Lassiter RB (2009) Quantification of Cry1Ac and Cry1F Bacillus thuringiensis insecticidal proteins in selected transgenic cotton plant tissue types. J Econ Entomol 102:1301–1308CrossRefGoogle Scholar
  51. Srinivasan MV (2010) Honey bees as a model for vision, perception, and cognition. Annu Rev Entomol 55:267–284CrossRefGoogle Scholar
  52. Stara J, Ourednickova J, Kocourek F (2011) Laboratory evaluation of the side effects of insecticides on Aphidius colemani (Hymenoptera: Aphidiidae), Aphidoletes aphidimyza (Diptera: Cecidomyiidae), and Neoseiulus cucumeris (Acari: Phytoseidae). J Pest Sci 84:25–31CrossRefGoogle Scholar
  53. Wratten SD, Gillespie M, Decourtye A, Mader E, Desneux N (2012) Pollinator habitat enhancement: benefits to other ecosystem services. Agr Ecosyst Environ doi: 10.1016/j.agee.2012.06.020
  54. Wu KM, Lu YH, Feng HQ, Jiang YY, Zhao JZ (2008) Suppression of cotton bollworm in multiple crops in China in areas with Bt toxin-containing cotton. Science 321:1676–1678CrossRefGoogle Scholar
  55. Yu HL, Li YH, Wu KM (2011) Risk assessment and ecological effects of transgenic Bacillus thuringiensis crops on non-target organisms. J Integr Plant Biol 53:520–538CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Peng Han
    • 1
    • 2
  • Chang-Ying Niu
    • 1
    Email author
  • Antonio Biondi
    • 2
    • 3
  • Nicolas Desneux
    • 2
  1. 1.Hubei Key Laboratory of Utilization of Insect Resources and Sustainable Control of Pests, College of Plant Science & TechnologyHuazhong Agricultural UniversityWuhanChina
  2. 2.French National Institute for Agricultural Research (INRA)Sophia-AntipolisFrance
  3. 3.Department of Agri-food and Environmental Systems ManagementUniversity of CataniaCataniaItaly

Personalised recommendations