Pesticides as a cause of honeybee (Apis mellifera) mortality and their persistence in honey
Abstract
Recently, there has been a widespread decline in honeybee (Apis mellifera) colonies globally, disrupting ecological balance and reducing the pollination capacity of many entomophilous plants. One of the primary causes of bee family deaths is the increasing use of pesticides, particularly insecticides, in agricultural practice. This study aimed to identify the causes of bee family mortality in various regions of Ukraine during 2021–2022 and to determine the breakdown potential of different pesticide groups in honey. Pesticide residues in biological samples were analyzed using liquid chromatography-mass spectrometry (UPLC-MS/MS) and gas chromatography-mass spectrometry (GC-MS/MS). In the bodies of dead bees, residues of various pesticides from different chemical groups, as well as their mixtures, were detected. In most cases, the cause of bee poisoning was mixtures of pyrethroids and neonicotinoids: thiamethoxam, clothianidin, and lambda-cyhalothrin; imidacloprid, lambda-cyhalothrin, and thiamethoxam; as well as clothianidin and lambda-cyhalothrin. The primary pesticides found in honey were neonicotinoids (58.8%), triazoles (29.6%), strobilurins (6.5%), and benzimidazoles (5.1%). Most pesticide levels did not exceed the maximum allowable levels in honey. The neonicotinoids detected in honey included thiacloprid, acetamiprid, imidacloprid, clothianidin, and thiamethoxam. The triazoles detected included tebuconazole, cyproconazole, flutriafol, and epoxiconazole; strobilurins included picoxystrobin, pyraclostrobin, and azoxystrobin; and benzimidazoles included carbendazim and thiophanate-methyl. Residues of neonicotinoids, triazoles, benzimidazoles, and strobilurins in honey did not undergo degradation over 12 months of storage at 4 °C. Storing honey at 20 °C after 12 months increased the degradation of thiacloprid by 21.2%, acetamiprid by 20.7%, and flutriafol by 36.3%. Between the 9th and 12th months at 20 °C, picoxystrobin concentration decreased by 24.5–38.0%, and carbendazim concentration decreased by 80.0% to a complete breakdown of residues in honey. The degradation of thiophanate-methyl in honey at 20 °C reached 28.0% by the 6th month, 46.0% by the 9th month, and 55.4% by the 12th month. This persistence of most pesticides in honey poses a significant risk of toxic effects on bee families as well as on human health.References
Ashraf, S. A., Mahmood, D., Elkhalifa, A. E. O., Siddiqui, A. J., Khan, M. I., Ashfaq, F., Patel, M., Snoussi, M., Kieliszek, M., & Adnan, M. (2023). Exposure to pesticide residues in honey and its potential cancer risk assessment. Food and Chemical Toxicology: an International Journal Published for the British Industrial Biological Research Association, 180, 114014.
Averill, A. L., Eitzer, B. D., & Drummond, F. A. (2024). Pesticide contamination in native North American crops. Part I – Development of a baseline and comparison of honey bee exposure to residues in lowbush blueberry and cranberry. Insects, 15(7), 489.
Bayer, E. V., Novozhitskaya, Y. N., Shevchenko, L. V., Mykhalska, V. M. (2017). Monitorynh zalyshkiv veterynarnykh preparativ u kharchovykh produktakh [Monitoring of residues of veterinary preparations in food products]. Ukrainian Journal of Ecology, 7(3), 251–257 (in Ukrainian).
Bayer, O. V., Yaremchuk, O. S., Yevtushenko, T. V., Shevchenko, L. V., Mykhalska, V. M., Dobrozhan, Y. V., Dovhopol, Y. V., & Varpikhovskyi, R. L. (2018). Rozrobka ta otsinka prydatnosti metodu vyznachennianitrofuraniv v mediza dopomohoiu ridynnoyi khromatohrafiyi vysokoho tysku – tandemnoyi mas-spektrometriyi (UPLC-RS-RS) [The development and validation of a rapid method for the determination of nitrofurans in honey using high pressure liquid chromatography tandem mass spectrometry (UPLC MS MS)]. Ukrainian Journal of Ecology, 8(1), 966–974 (in Ukrainian).
Chiaia-Hernandez, A. C., Keller, A., Wächter, D., Steinlin, C., Camenzuli, L., Hollender, J., & Krauss, M. (2017). Long-term persistence of pesticides and TPS in archived agricultural soil samples and comparison with pesticide application. Environmental Science and Technology, 51(18), 10642–10651.
Chmiel, J. A., Daisley, B. A., Pitek, A. P., Thompson, G. J., & Reid, G. (2020). Understanding the effects of sublethal pesticide exposure on honey bees: A role for probiotics as mediators of environmental stress. Frontiers in Ecology and Evolution, 8, 22.
Christen, V. (2023). Different effects of pesticides on transcripts of the endocrine regulation and energy metabolism in honeybee foragers from different colonies. Scientific Reports, 13, 1985.
Crall, J. D., & Raine, N. E. (2023). How do neonicotinoids affect social bees? Linking proximate mechanisms to ecological impacts. Advances in Insect Physiology, 64, 191–253.
Dukhnytskyi, V. B., Horalskyi, L. P., Sokolyuk, V. M., Gutyj, B. V., Ishchenko, V. D., Ligomina, І. P., Kolesnik, N. L., & Dzhmil, V. I. (2024). Morphofunctional changes in the internal organs of laying hens affected by chronic thiamethoxam intoxication. Regulatory Mechanisms in Biosystems, 15(3), 578–586.
Faly, L. I., Brygadyrenko, V. V., Orzekauskaite, A., & Paulauskas, A. (2023). Sensitivity of non-target groups of invertebrates to cypermethrin. Biosystems Diversity, 31(3), 393–400.
Faly, L. I., Brygadyrenko, V. V., Orzekauskaite, A., & Paulauskas, A. (2023). Sensitivity of nontarget groups of invertebrates to cypermethrin. Biosystems Diversity, 31(3), 393–400.
Faly, L., & Brygadyrenko, V. (2024). Effects of pirimiphos-methyl on non-target invertebrates. Biology, 13(10), 823.
Froger, C., Jolivet, C., Budzinski, H., Pierdet, M., Caria, G., Saby, N. P. A., Arrouays, D., & Bispo, A. (2023). Pesticide residues in french soils: occurrence, risks, and persistence. Environmental Science & Technology, 57(20), 7818–7827.
Gandara, L., Jacoby, R., Laurent, F., Spatuzzi, M., Vlachopoulos, N., Borst, N. O., Ekmen, G., Potel, C. M., Garrido-Rodriguez, M., Böhmert, A. L., Misunou, N., Bartmanski, B. J., Li, X. C., Kutra, D., Hériché, J. K., Tischer, C., Zimmermann-Kogadeeva, M., Ingham, V. A., Savitski, M. M., Masson, J. B., Zimmermann, M., & Crocker, J. (2024). Pervasive sublethal effects of agrochemicals on insects at environmentally relevant concentrations. Science, 386(6720), 446–453.
Graham, K. K., Milbrath, M. O., Zhang, Y., Soehnlen, A., Baert, N., McArt, S., & Isaacs, R. (2021). Identities, concentrations, and sources of pesticide exposure in pollen collected by managed bees during blueberry pollination. Scientific Reports, 11(1), 16857.
Haas, J., Zaworra, M., Glaubitz, J., Hertlein, G., Kohler, M., Lagojda, A., Lueke, B., Maus, C., Almanza, M. T., Davies, T. G. E., Bass, C., & Nauen, R. (2021). A toxicogenomics approach reveals characteristics supporting the honey bee (Apis mellifera L.) safety profile of the butenolide insecticide flupyradifurone. Ecotoxicology and Environmental Safety, 217, 112247.
Hisamoto, S., Ikegami, M., Goka, K., & Sakamoto, Y. (2024). The impact of landscape structure on pesticide exposure to honey bees. Nature Communications, 15(1), 8999.
Hrynko, I., Kaczyński, P., & Łozowicka, B. (2021). A global study of pesticides in bees: QuEChERS as a sample preparation methodology for their analysis – Critical review and perspective. Science of the Total Environment, 792, 148385.
Jeschke, P. (2024). Recent developments in fluorine-containing pesticides. Pest Management Science, 80, 3065–3087.
Kadlikova, K., Vaclavikova, M., Halesova, T., Kamler, M., Markovic, M., & Erban, T. (2021). The investigation of honey bee pesticide poisoning incidents in Czechia. Chemosphere, 263, 128056.
Khalifa, S. A. M., Elshafiey, E. H., Shetaia, A. A., El-Wahed, A. A. A., Algethami, A. F., Musharraf, S. G., AlAjmi, M. F., Zhao, C., Masry, S. H. D., Abdel-Daim, M. M., Halabi, M. F., Kai, G., Al Naggar, Y., Bishr, M., Diab, M. A. M., & El-Seedi, H. R. (2021). Overview of bee pollination and its economic value for crop production. Insects, 12(8), 688.
Knuth, D., Gai, L., Silva, V., Harkes, P., Hofman, J., Šudoma, M., Bílková, Z., Alaoui, A., Mandrioli, D., Pasković, I., Pasković, M. P., Baldi, I., Bureau, M., Alcon, F., Contreras, J., Glavan, M., Abrantes, N., Campos, I., Norgaard, T., Lwanga, E. H., Scheepers, P. T. J., Ritsema, C. J., & Geissen, V. (2024). Pesticide residues in organic and conventional agricultural soils across Europe: Measured and predicted concentrations. Environmental Science and Technology, 58(15), 6744–6752.
Kozak, V. M., Romanenko, E. R., & Brygadyrenko, V. V. (2020). Influence of herbicides, insecticides and fungicides on food consumption and body weight of Rossiulus kessleri (Diplopoda, Julidae). Biosystems Diversity, 28(3), 272–280.
Lamas, M., Rodrigues, F., Amaral, M. H., Delerue-Matos, C., & Fernandes, V. C. (2023). Contaminant cocktails of high concern in honey: Challenges, QuEChERS extraction and levels. Separations, 10(2), 142.
Leska, A., Nowak, A., Nowak, I., & Górczyńska, A. (2021). Effects of Insecticides and microbiological contaminants on Apis mellifera health. Molecules, 26(16), 5080.
Main, A. R., Hladik, M. L., Webb, E. B., Goyne, K. W., & Mengel, D. (2020). Beyond neonicotinoids – wild pollinators are exposed to a range of pesticides while foraging in agroecosystems. Science of the Total Environment, 742, 140436.
Mishra, C. S. K., Samal, S., & Samal, R. R. (2022). Evaluating earthworms as candidates for remediating pesticide contaminated agricultural soil: A review. Frontiers in Environmental Science, 10, 1924480.
Omelchun, Y. A., Kobish, A. I., Klochkova, N. P., & Shevchenko, L. V. (2022). Validation of the multiresidue method analysis for pesticides in bee honey by UPLC-MS/MS using the method of samples preparation QuEChERS. Methods and Objects of Chemical Analysis, 17(3), 141–152.
Omelchun, Y., Shevchenko, L., Voynalovich, M., Savchenko, O., Hryshchenco, N., Tkach, G., Androshchuk, O., Drachuk, O., Kozii, M., Rzhevskyi, H., & Slyva, Y. (2023). Effects of pesticides on bee populations and safety of bee honey in Ukraine. Potravinarstvo, 17, 801–815.
Ostiguy, N., Drummond, F. A., Aronstein, K., Eitzer, B., Ellis, J. D., Spivak, M., & Sheppard, W. S. (2019). Honey bee exposure to pesticides: A four-year nationwide study. Insects, 10(1), 13.
Pashte, V. V., & Patil, C. S. (2018). Toxicity and poisoning symptoms of selected insecticides to honey bees (Apis mellifera mellifera L.). Archives of Biological Sciences, 70(1), 5–12.
Pathak, V. M., Verma, V. K., Rawat, B. S., Kaur, B., Babu, N., Sharma, A., Dewali, S., Yadav, M., Kumari, R., Singh, S., Mohapatra, A., Pandey, V., Rana, N., & Cunill, J. M. (2022). Current status of pesticide effects on environment, human health and it's eco-friendly management as bioremediation: A comprehensive review. Frontiers in Microbiology, 13, 962619.
Prado, A., Pioz, M., Vidau, C., Requier, F., Jury, M., Crauser, D., Brunet, J. L., Le Conte, Y., & Alaux, C. (2019). Exposure to pollen-bound pesticide mixtures induces longer-lived but less efficient honey bees. Science of the Total Environment, 650(1), 1250–1260.
Raine, N. E., & Rundlöf, M. (2024). Pesticide exposure and effects on non-Apis bees. Annual Review of Entomology, 69, 551–576.
Ramirez-Cando, L. J., Guzmán-Vallejos, M. S., Aguayo, L. G., Vera-Erazo, F. D., & Ballaz, S. J. (2023). Neurocytotoxicity of imidacloprid- and acetamiprid-based comercial insecticides over the differentiation of SH-SY5Y neuroblastoma cells. Heliyon, 9(5), e15840.
Rondeau, S., & Raine, N. E. (2024). Single and combined exposure to ‘bee safe’ pesticides alter behaviour and offspring production in a ground-nesting solitary bee (Xenoglossa pruinosa). Proceedings of the Royal Society B, 291, 0232939.
Schuhmann, A., Schmid, A. P., Manzer, S., Schulte, J., & Scheiner, R. (2022). Interaction of insecticides and fungicides in bees. Frontiers in Insect Science, 1, 808335.
Shi, M., Guo, Y., Wu, Y. Y., Dai, P. L., Dai, S. J., Diao, Q. Y., & Gao, J. (2023). Acute and chronic effects of sublethal neonicotinoid thiacloprid to Asian honey bee (Apis cerana cerana). Pesticide Biochemistry and Physiology, 194, 105483.
Wang, Y. Z., Cao, C. Q., & Wang, D. (2022). Physiological responses of the firefly Pyrocoelia analis (Coleoptera: Lampyridae) to an environmental residue from chemical pesticide imidacloprid. Frontiers in Physiology, 13, 879216.
Yang, Y., Wu, Y., Long, H., Ma, X., Shariati, K., Webb, J., Guo, L., Pan, Y., Ma, M., Deng, C., Cao P., & Chen, J. (2023). Global honeybee health decline factors and potential conservation techniques. Food Security, 15, 855–875.
Zattara, E. E., & Aizen, M. A. (2021). Worldwide occurrence records suggest a global decline in bee species richness. One Earth, 4(1), 114–123.
Zhao, H., Li, G., Cui, X., Wang, H., Liu, Z., Yang, Y., & Xu, B. (2022). Review on effects of some insecticides on honey bee health. Pesticide Biochemistry and Physiology, 188, 105219.
Zioga, E., White, B., & Stout, J. C. (2023). Honey bees and bumble bees may be exposed to pesticides differently when foraging on agricultural areas. Science of the Total Environment, 896, 166214.
Zuščíková, L., Bažány, D., Greifová, H., Knížatová, N., Kováčik, A., Lukáč, N., & Jambor, T. (2023). Screening of toxic effects of neonicotinoid insecticides with a focus on acetamiprid: A review. Toxics, 11(7), 598.



