, Volume 24, Issue 1, pp 130–142 | Cite as

Lethal and sublethal effects of azadirachtin on the bumblebee Bombus terrestris (Hymenoptera: Apidae)

  • Wagner Faria Barbosa
  • Laurens De Meyer
  • Raul Narciso C. Guedes
  • Guy SmaggheEmail author


Azadirachtin is a biorational insecticide commonly reported as selective to a range of beneficial insects. Nonetheless, only few studies have been carried out with pollinators, usually emphasizing the honeybee Apis mellifera and neglecting other important pollinator species such as the bumblebee Bombus terrestris. Here, lethal and sublethal effects of azadirachtin were studied on B. terrestris via oral exposure in the laboratory to bring out the potential risks of the compound to this important pollinator. The compound was tested at different concentrations above and below the maximum concentration that is used in the field (32 mg L−1). As most important results, azadirachtin repelled bumblebee workers in a concentration-dependent manner. The median repellence concentration (RC50) was estimated as 504 mg L−1. Microcolonies chronically exposed to azadirachtin via treated sugar water during 11 weeks in the laboratory exhibited a high mortality ranging from 32 to 100 % with a range of concentrations between 3.2 and 320 mg L−1. Moreover, no reproduction was scored when concentrations were higher than 3.2 mg L−1. At 3.2 mg L−1, azadirachtin significantly inhibited the egg-laying and, consequently, the production of drones during 6 weeks. Ovarian length decreased with the increase of the azadirachtin concentration. When azadirachtin was tested under an experimental setup in the laboratory where bumblebees need to forage for food, the sublethal effects were stronger as the numbers of drones were reduced already with a concentration of 0.64 mg L−1. Besides, a negative correlation was found between the body mass of male offspring and azadirachtin concentration. In conclusion, our results as performed in the laboratory demonstrated that azadirachtin can affect B. terrestris with a range of sublethal effects. Taking into account that sublethal effects are as important as lethal effects for the development and survival of the colonies of B. terrestris, this study confirms the need to test compounds on their safety, especially when they have to perform complex tasks such as foraging. The latter agrees with the recent European Food Safety Authority guidelines to assess ‘potentially deleterious’ compounds for sublethal effects on behavior.

Graphical Abstract


Bumblebee Chronic oral exposure Insect growth regulator Neem Repellence effect Reproduction 



We thank the CAPES Foundation (Brazilian Ministry of Education) for the scholarship and financial support to W.F. Barbosa. This project was also support by the Research Council of Ghent University (BOF-UGent) and the Flemish government agency for Innovation by Science and Technology (IWT, Belgium).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Allison PD (1998) Survival analysis using SAS: a practical guide. SAS Institute Inc., CaryGoogle Scholar
  2. Alves JE (2010) Toxicidade do nim (Azadirachta indica A. Juss.: Meliaceae) para Apis mellifera e sua importância apícola na Caatinga e Mata Litorânea cearense. Thesis, Universidade Federal do CearáGoogle Scholar
  3. Amin MR, Bussière LF, Goulson D (2012) Effects of male age and size on mating success in the bumblebee Bombus terrestris. Insect Behav 25:362–374CrossRefGoogle Scholar
  4. 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
  5. Barnby MA, Klocke JA (1990) Effects of azadirachtin on levels of ecdysteroids and prothoracicotropic hormone-like activity in Heliothis virescens (Fabr.) larvae. J Insect Physiol 36(125):131Google Scholar
  6. Blacquiere T, Smagghe G, Van Gestel CAM, Mommaerts V (2012) Neonicotinoids in bees: a review on concentrations, side-effects and risk-assessment. Ecotoxicology 21:973–992CrossRefGoogle Scholar
  7. Blaney WM, Simmonds MSJ, Ley WV, Anderson JC, Toogood PL (1990) Antifeedant effects of azadirachtin and structurally related compounds on lepidopterous larvae. Entomol Exp Appl 55:149–160CrossRefGoogle Scholar
  8. Bloch G, Borst DW, Huang Z-Y, Robinson GE, Cnaani J, Hefetz A (2000a) Juvenile hormone titers, juvenile hormone biosynthesis, ovarian development and social environment in Bombus terrestris. J Insect Physiol 46:47–57CrossRefGoogle Scholar
  9. Bloch G, Hefetz A, Hartfelder K (2000b) Ecdysteroid titer, ovary status, and dominance in adult worker and queen bumble bees (Bombus terrestris). J Insect Physiol 46:1033–1040CrossRefGoogle Scholar
  10. Boeke SJ, Boersma MG, Alink GM, van Loona JJA, van Huis A, Dicke M, Rietjens IMCM (2004) Safety evaluation of neem (Azadirachta indica) derived pesticides. J Ethnopharmacol 94:25–41CrossRefGoogle Scholar
  11. Brittain C, Potts SG (2011) The potential impacts of insecticides on the life-history traits of bees and the consequences for pollination. Basic Appl Ecol 12:321–331CrossRefGoogle Scholar
  12. Brown MJF, Paxton RJ (2009) The conservation of bees: a global perspective. Apidologie 40:410–416CrossRefGoogle Scholar
  13. Bryden J, Gill RJ, Mitton RAA, Raine NE, Jansen VAA (2013) Chronic sublethal stress causes bee colony failure. Ecol Lett 16:1463–1469CrossRefGoogle Scholar
  14. Cantrell CL, Dayan FE, Duke SO (2012) Natural products as sources for new pesticides. J Natural Prod 75:1231–1242CrossRefGoogle Scholar
  15. Cassida JE, Quistad GB (2004) Why insecticides are more toxic to insects than people: the unique toxicology of insects. J Pestic Sci 29:81–86CrossRefGoogle Scholar
  16. Chen Y, Ruberson JR (2008) Starvation effects on larval development of beet armyworm (Lepidoptera: Noctuidae). J Entomol Sci 43:247–253Google Scholar
  17. Cooper J, Dobson H (2007) The benefits of pesticides to mankind and the environment. Crop Prot 26:1337–1348CrossRefGoogle Scholar
  18. Decourtye A, Armengaud C, Renou M, Devillers J, Cluzeau S, Gauthier M, Pham-Delegue MH (2004) Imidacloprid impairs memory and brain metabolism in the honeybee (Apis mellifera L.). Pestic Biochem Physiol 78:83–92CrossRefGoogle Scholar
  19. Dorn A, Rademacher JM, Sehn E (1986) Effects of azadirachtin on the moulting cycle, endocrine system and ovaries in last-instar larvae of the milkweed bug, Oncopeltus fasciatus. J Insect Physiol 32:231–238CrossRefGoogle Scholar
  20. European Food Safety Authority (2012) Scientific opinion on the science behind the development of a risk assessment of plant protection products on bees (Apis mellifera, Bombus spp. and solitary bees). EFSA J 10:2668Google Scholar
  21. Freitas BM, Imperatriz-Fonseca VL, Medina LM, Kleinert ADP, Galetto L, Nates-Parra G, Quezada-Euan JJG (2009) Diversity, threats and conservation of native bees in the Neotropics. Apidologie 40:332–346CrossRefGoogle Scholar
  22. Gallai N, Salles JM, Settele J, Vaissiere BE (2009) Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Ecol Econ 68:810–821CrossRefGoogle Scholar
  23. Gels JA, Held DW, Potter DA (2002) Hazards of insecticides to the bumble bees Bombus impatiens (Hymenoptera: Apidae) foraging on flowering white clover in turf. J Econ Entomol 95:722–728CrossRefGoogle Scholar
  24. Gill RJ, Ramos-Rodriguez O, Raine NE (2012) Combined pesticide exposure severely affects individual- and colony-level traits in bees. Nature 491:105–119 U119CrossRefGoogle Scholar
  25. Goulson D, Lye GC, Darvill B (2008) Decline and conservation of bumble bees. Annu Rev Entomol 53:191–208CrossRefGoogle Scholar
  26. Isman MB (2006) Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world. Annu Rev Entomol 51:45–66CrossRefGoogle Scholar
  27. Jeschke P, Nauen R (2008) Neonicotinoids—from zero to hero in insecticide chemistry. Pest Manag Sci 64:1084–1098CrossRefGoogle Scholar
  28. Johnson RM, Dahlgren L, Siegfried BD, Ellis MD (2013) Acaricide, fungicide and drug interactions in honey bees (Apis mellifera). PLoS ONE 8:e54092CrossRefGoogle Scholar
  29. Klein AM, Vaissiere BE, Cane JH, Steffan-Dewenter I, Cunningham SA, Kremen C, Tscharntke T (2007) Importance of pollinators in changing landscapes for world crops. Proc R Soc B 274:303–313CrossRefGoogle Scholar
  30. Kovacova J, Hrbek V, Kloutvorova J, Kocourek V, Drabova L, Hajslova J (2013) Assessment of pesticide residues in strawberries grown under various treatment regimes. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 30:2123–2135CrossRefGoogle Scholar
  31. Kumar P, Poehling H-M (2006) Persistence of soil and foliar azadirachtin treatments to control sweetpotato whitefly Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) on tomatoes under controlled (laboratory) and field (netted greenhouse) conditions in the humid tropics. J Pest Sci 79:189–199CrossRefGoogle Scholar
  32. Lamberth C, Jeanmart S, Luksch T, Plant A (2013) Current challenges and trends in the discovery of agrochemicals. Science 341:742–746CrossRefGoogle Scholar
  33. Lowery DT, Bellerose S, Smirle MJ, Vincent C, Pilon JG (1996) Effect of neem on the growth and development of the obliquebanded leafroller, Choristoneura rosaceana. Entomol Exp Appl 79:203–209CrossRefGoogle Scholar
  34. Lucantoni L, Giusti F, Cristofaro M, Pasqualini L, Esposito F, Lupetti P, Habluetzel A (2006) Effects of a neem extract on blood feeding, oviposition and oocyte ultrastructure in Anopheles stephensi Liston (Diptera: Culicidae). Tissue Cell 38:361–371CrossRefGoogle Scholar
  35. Melathopoulos AP, Winston ML, Whittington R, Smith T, Lindberg C, Mukai A, Moore M (2000) Comparative laboratory toxicity of neem pesticides to honey bees (Hymenoptera: Apidae), their mite parasites Varroa jacobsoni (Acari: Varroidae) and Acarapis woodi (Acari: Tarsonemidae), and brood pathogens Paenibacillus larvae and Ascophaera apis. J Econ Entomol 93:199–209CrossRefGoogle Scholar
  36. Metcalf RL (1980) Changing role of insecticides in crop protection. Annu Rev Entomol 25:219–255CrossRefGoogle Scholar
  37. Michener CD (1974) The social behaviour of the bees. Harvard University Press, CambridgeGoogle Scholar
  38. Mommaerts V, Reynders S, Boulet J, Besard L, Sterk G, Smagghe G (2010) Risk assessment for side-effects of neonicotinoids against bumblebees with and without impairing foraging behavior. Ecotoxicology 19:207–215CrossRefGoogle Scholar
  39. Mordue (Luntz) AJ, Nisbet AJ (2000) Azadirachtin from the neem tree Azadirachta indica: its action against insects. An Soc Entomol Brasil 29:615–632Google Scholar
  40. Mordue (Luntz) AJ, Simmonds MSJ, Ley SV, Blaney WM, Mordue W, Nasiruddin M, Nisbet AJ (1998) Actions of azadirachtin, a plant allelochemical, against insects. Pest Sci 54:277–284Google Scholar
  41. Morgan ED (2009) Azadirachtin, a scientific gold mine. Bioorgan Med Chem 17:4096–4105CrossRefGoogle Scholar
  42. Munyiri FN, Asano W, Shintani Y, Ishikawa Y (2003) Threshold weight for starvation-triggered metamorphosis in the yellow-spotted longicorn beetle, Psacothea hilaris (Coleoptera: Cerambycidae). Appl Entomol Zool 38:509–515CrossRefGoogle Scholar
  43. Nathan SS, Choi MY, Paik CH, Seo HY, Kim JD, Kang SM (2007) The toxic effects of neem extract and azadirachtin on the brown planthopper, Nilaparvata lugens (Stal) (BPH) (Homoptera: Delphacidae). Chemosphere 67:80–88CrossRefGoogle Scholar
  44. Naumann K, Isman MB (1996) Toxicity of a neem (Azadirachta indica A. Juss) insecticide to larval honey bees. Am Bee J 136:518–520Google Scholar
  45. Salehzadeh A, Jabbar A, Jennens L, Ley SV, Annadurai RS, Adams R, Strang RH (2002) The effects of phytochemical pesticides on the growth of cultured invertebrate and vertebrate cells. Pest Manag Sci 58:268–276CrossRefGoogle Scholar
  46. Sayah F (2002) Ultrastructural changes in the corpus allatum after azadirachtin and 20-hydroxyecdysone treatment in adult females of Labidura riparia (Dermaptera). Tissue Cell 34:53–62CrossRefGoogle Scholar
  47. Sayah F, Fayet C, Idaomar M, Karlinsky A (1996) Effects of azadirachtin on vitellogenesis of Labidura riparia (Insecta: Dermaptera). Tissue Cell 28:741–749CrossRefGoogle Scholar
  48. Sayah F, Idaomar M, Soranzo L, Karlinsky A (1998) Endocrine and neuroendocrine effects of azadirachtin in adult females of the earwig Labidura riparia. Tissue Cell 30:86–94CrossRefGoogle Scholar
  49. Schluns H, Schluns EA, van Praagh J, Moritz RFA (2003) Sperm numbers in drone honeybees (Apis mellifera) depend on body size. Apidologie 34:577–584CrossRefGoogle Scholar
  50. Simmonds MSJ, Blaney WM, Ley SV, Anderson JC, Banteli R, Denholm AA, Green PCW, Grossman RB, Gutteridge C, Jennens L, Smith SC, Toogood PL, Wood A (1995) Behavioral and neurophysiological responses of Spodoptera littoralis to azadirachtin and a range of synthetic analogs. Entomol Exp Appl 77:69–80CrossRefGoogle Scholar
  51. Smagghe G, Deknopper J, Meeus I, Mommaerts V (2013) Dietary chlorantraniliprole suppresses reproduction in worker bumblebees. Pest Manag Sci 69:787–791CrossRefGoogle Scholar
  52. Thoeming G, Poehling HM (2006) Soil application of different neem products to control Ceratothripoides claratris (Thysanoptera: Thripidae) on tomatoes grown under protected cultivation in the humid tropics (Thailand). Int J Pest Manage 52:239–248CrossRefGoogle Scholar
  53. Thompson DG, Kreutzweiser DP, Staznik B, Chartrand D, Capell S (2002) Fate and persistence of azadirachtin a following applications to mesocosms in a small forest lake. Bull Environ Contam Toxicol 69:250–256CrossRefGoogle Scholar
  54. Thompson HM, Wilkins S, Battersby AH, Waite RJ, Wilkinson D (2005) The effects of four insect growth-regulating (IGR) insecticides on honeybee (Apis mellifera L.) colony development, queen rearing and drone sperm production. Ecotoxicology 14:757–769CrossRefGoogle Scholar
  55. Timmins WA, Reynolds SF (1992) Azadirachtin inhibits secretion of trypsin in midgut of Manduca sexta caterpillars: reduced growth due to impaired protein digestion. Entomol Exp Appl 63:47–54CrossRefGoogle Scholar
  56. Trumm P, Dorn A (2000) Effects of azadirachtin on the regulation of midgut peristalsis by the stomatogastric nervous system in Locusta migratoria. Phytoparasitica 28:7–26CrossRefGoogle Scholar
  57. van Doom A, Heringa J (1986) The ontogeny of a dominance hierarchy in colonies of the bumble bee Bombus terrestris (Hymenoptera: Apidae). Insect Soc 33:3–25CrossRefGoogle Scholar
  58. van Honk CGJ, Roseler PF, Hoogeveen JC (1981) Factors influencing the egg laying of workers in a captive colony Bombus terrestris. Behav Ecol Sociobiol 9:9–14CrossRefGoogle Scholar
  59. vanEngelsdorp D, Meixner MD (2010) A historical review of managed bee populations in Europe and United States and the factors that may affect them. J Invertebr Pathol 103:S80–S95CrossRefGoogle Scholar
  60. Velthuis HHW, van Doorn A (2006) A century of advances in bumblebee domestication and the economic and environmental aspects of its commercialization for pollination. Apidologie 37:421–451CrossRefGoogle Scholar
  61. Villaverde JJ, Sevilla-Moran B, Sandin-Espana P, Lopez-Goti C, Alonso-Prados JL (2014) Biopesticides in the framework of the European Pesticide Regulation (EC) No. 1107/2009. Pest Manag Sci 70:2–5CrossRefGoogle Scholar
  62. Wu JY, Anelli CM, Sheppard WS (2011) Sub-lethal effects of pesticide residues in brood comb on worker honey bee (Apis mellifera) development and longevity. PLoS ONE 6:e14720CrossRefGoogle Scholar
  63. Yang EC, Chuang YC, Chen YL, Chang LH (2008) Abnormal foraging behavior induced by sublethal dosage of imidacloprid in the honey bee (Hymenoptera: Apidae). J Econ Entomol 101:1743–1748CrossRefGoogle Scholar
  64. Zanuncio JC, Molina-Rugama AJ, Santos GP, Ramalho FD (2002) Effect of body weight on fecundity and longevity of the stinkbug predator Podisus rostralis. Pesqui Agropecu Bras 37:1225–1230CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Wagner Faria Barbosa
    • 1
    • 2
  • Laurens De Meyer
    • 1
  • Raul Narciso C. Guedes
    • 2
  • Guy Smagghe
    • 1
    Email author
  1. 1.Department of Crop Protection, Faculty of Bioscience EngineeringGhent UniversityGhentBelgium
  2. 2.Departamento de EntomologiaUniversidade Federal de ViçosaViçosaBrazil

Personalised recommendations