In silico evaluation of toxic effects of benalaxyl-m on bees

Abstract

This study evaluated the acute toxic effects of Benalaxyl-M on honey bees (Apis mellifera) using BeeToxAI, a machine learning-based QSAR (Quantitative Structure-Activity Relationship) tool, through in silico methods. The analysis results indicated that Benalaxyl-M is non-toxic in terms of acute oral and contact toxicity for honey bees (83-86% reliability). The Applicability Domain analysis confirmed the reliability of the predictions, while contribution maps identified functional groups contributing to toxicity. Although the findings suggest a low acute risk of Benalaxyl-M to honey bees, experimental validation is recommended. This study highlights the potential of AI-based tools in the ecotoxicological risk assessment of pesticides.

Keywords

• Benalaxyl-M
• honey bees
• ecotoxicology
• in silico modeling
• QSAR
• BeeToxAI
• pesticide risk assessment

Benalaxyl-M Maddesinin Arılar Üzerindeki Toksik Etkilerinin In Silico Değerlendirilmesi

Öz

Bu çalışmada, Benalaxyl-M maddesinin bal arıları (Apis mellifera) üzerindeki akut toksik etkileri, makine öğrenimi tabanlı bir QSAR (Kantitatif Yapı-Aktivite İlişkisi) aracı olan BeeToxAI kullanılarak in silico yöntemlerle değerlendirilmiştir. Analiz sonuçları, Benalaxyl-M'nin bal arıları için akut oral ve kontakt toksisite açısından toksik olmadığını (%83-86 güvenilirlik) göstermiştir. Uygulanabilirlik Alanı analizi, tahminlerin güvenilir olduğunu doğrularken, katkı haritaları moleküldeki toksisiteye katkı yapan fonksiyonel grupları belirlemede kullanılmıştır. Elde edilen bulgular, Benalaxyl-M'nin bal arıları üzerindeki düşük akut riskini ortaya koysa da, sonuçların deneysel çalışmalarla desteklenmesi önerilmektedir. Bu çalışma, pestisitlerin ekotoksikolojik risk değerlendirmelerinde yapay zeka tabanlı araçların potansiyelini vurgulamaktadır.

Anahtar Kelimeler

• Benalaxyl-M
• bal arıları
• ekotoksikoloji
• in silico modelleme
• QSAR
• BeeToxAI
• pestisit risk değerlendirmesi

Kaynakça

1. M. Tudi, H. D. Ruan, L. Wang, J. Lyu, R. Sadler, D. Connell, C. Chu, and D. T. Phung, "Agriculture development, pesticide application and its impact on the environment," Int. J. Environ. Res. Public Health, vol. 18, no. 3, p. 1112, Jan. 2021, doi: 10.3390/ijerph18031112.
2. W. Zhou, M. Li, and V. Achal, "A comprehensive review on environmental and human health impacts of chemical pesticide usage," Emerging Contaminants, vol. 11, no. 1, p. 100410, 2025, doi: 10.1016/j.emcon.2024.100410.
3. M. F. Ahmad et al., "Pesticides impacts on human health and the environment with their mechanisms of action and possible countermeasures," Heliyon, vol. 10, no. 7, p. e29128, 2024, doi: 10.1016/j.heliyon.2024.e29128.
4. M. Lechenet, G. Py, F. Dessaint, and D. Makowski, "Reducing pesticide use while preserving crop productivity and profitability on arable farms," Nature Plants, vol. 3, Art. no. 17008, 2017, doi: 10.1038/nplants.2017.8.
5. C. Shekhar et al., "A systematic review of pesticide exposure, associated risks, and long-term human health impacts," Toxicol. Rep., vol. 13, p. 101840, 2024, doi: 10.1016/j.toxrep.2024.101840.
6. E. Hakme, M. Poulsen, and A. Lassen, "A Comprehensive Review on Pesticide Residues in Human Urine," Journal of Agricultural and Food Chemistry, 2024. [Online]. Available: https://pubs.acs.org/doi/abs/10.1021/acs.jafc.4c02705.
7. R. M. González-Rodríguez, B. Cancho-Grande, and J. Simal-Gándara, "Decay of fungicide residues during vinification of white grapes harvested after the application of some new active substances against downy mildew," Food Chem., vol. 125, no. 2, pp. 549-560, Mar. 2011, doi: 10.1016/j.foodchem.2010.09.047.
8. European Food Safety Authority (EFSA) et al., "Review of the existing maximum residue levels for benalaxyl-M according to Article 12 of Regulation (EC) No 396/2005," EFSA J., vol. 17, no. 7, p. 5818, Jul. 2019, doi: 10.2903/j.efsa.2019.5818.

9. J. G. Zaller and C. A. Brühl, "Editorial: Non-target effects of pesticides on organisms inhabiting agroecosystems," Front. Environ. Sci., vol. 7, Art. no. 75, 2019, doi: 10.3389/fenvs.2019.00075. [ J. Grassl et al., "Synergistic effects of pathogen and pesticide exposure on honey bee (Apis mellifera) survival and immunity," J. Invertebr. Pathol., vol. 159, pp. 78-86, 2018, doi: 10.1016/j.jip.2018.10.005.
10. F. Sanchez-Bayo and K. Goka, "Pesticide residues and bees- A risk assessment," PLoS ONE, 2014. https://doi.org/10.1371/journal.pone.0094482.
11. M. Schott et al., "Temporal dynamics of whole body residues of the neonicotinoid insecticide imidacloprid in live or dead honeybees," Sci. Rep., vol. 7, p. 6288, 2017, doi: 10.1038/s41598-017-06259-z.
12. V. V. Kleandrova et al., "In silico assessment of the acute toxicity of chemicals: Recent advances and new model for multitasking prediction of toxic effect," Mini Rev. Med. Chem., vol. 15, no. 8, pp. 677-686, 2015, doi: 10.2174/1389557515666150219143604.
13. R. M. González-Rodríguez, B. Cancho-Grande, and J. Simal-Gándara, "Multiresidue determination of 11 new fungicides in grapes and wines by liquid-liquid extraction/clean-up and programmable temperature vaporization injection with analyte protectants/gas chromatography/ion trap mass spectrometry," J. Chromatogr. A, vol. 1216, no. 32, pp. 6033-6042, Aug. 2009, doi: 10.1016/j.chroma.2009.06.046.
14. PubChem, “Benalaxyl-M,” National Center for Biotechnology Information. [Online]. Available: https://pubchem.ncbi.nlm.nih.gov/compound/176648. [Accessed: May 2, 2025].
15. J. T. Moreira-Filho et al., "BeeToxAI: An artificial intelligence-based web app to assess acute toxicity of chemicals to honey bees," Artif. Intell. Life Sci., vol. 1, Dec. 2021, doi: 10.1016/j.ailsci.2021.100013.
16. F. Wang et al., "Graph attention convolutional neural network model for chemical poisoning of honey bees' prediction," Sci. Bull., vol. 65, no. 14, pp. 1184-1191, 2020, doi: 10.1016/j.scib.2020.04.006.
17. S. Riniker and G. A. Landrum, "Similarity maps-a visualization strategy for molecular fingerprints and machine-learning methods," J. Cheminf., vol. 5, pp. 1-7, 2013, doi: 10.1186/1758-2946-5-43.
18. BeeToxAI. [Online]. Available: http://beetoxai.labmol.com.br. Accessed: Apr. 10, 2025.
19. S. A. Mansour, "Pesticide exposure-Egyptian scene," Toxicology, vol. 198, no. 1-3, pp. 91-115, 2004, https://doi.org/10.1016/j.tox.2004.01.036
20. Tarım İlacı (Pestisit) Kullanımı, Ankara, Turkey: Tarım ve Orman Bakanlığı, 2023. [Online]. Available: https://cevreselgostergeler.csb.gov.tr/tarim-ilaci-pestisit-kullanimi-i-85834. Accessed: Apr. 25, 2025.
21. N. Vural, Toksikoloji, Ankara, Turkey: Ankara Üniversitesi Eczacılık Fakültesi Yayınları, 2005, p. 659.
22. S. Ali et al., "Environmental and Health Effects of Pesticide Residues," in Sustainable Agriculture Reviews 48, I. Inamuddin, M. I. Ahamed, and E. Lichtfouse, Eds. Cham, Switzerland: Springer, 2021, ch. 8. [Online]. Available: https://doi.org/10.1007/978-3-030-54719-6_8.
23. M. A. Beketov, B. J. Kefford, R. B. Schäfer, and M. Liess, "Pesticides reduce regional biodiversity of stream invertebrates," Proc. Natl. Acad. Sci. U.S.A., vol. 110, no. 27, pp. 11039–11043, 2013, doi: 10.1073/pnas.1305618110.
24. G. L. Baron et al., "Pesticide reduces bumblebee colony initiation and increases probability of population extinction," Nat. Ecol. Evol., vol. 1, pp. 1308-1316, 2017, doi: 10.1038/s41559-017-0260-1.
25. T. Haarmann, M. Spivak, D. Weaver, B. Weaver, and T. Glenn, “Effects of fluvalinate and coumaphos on queen honey bees (Hymenoptera: Apidae) in two commercial queen rearing operations,” Journal of Economic Entomology, vol. 95, no. 1, pp. 28–35, Feb. 2002., https://doi.org/10.1603/0022-0493-95.1.28
26. S. Tosi, J. C. Nieh, F. Sgolastra, R. Cabbri, and P. Medrzycki, “Neonicotinoid pesticides and nutritional stress synergistically reduce survival in honey bees,” Proc. R. Soc. B, vol. 284, no. 1869, p. 20171711, 2017. http://doi.org/10.1098/rspb.2017.1711
27. V. L. Imperatriz-Fonseca et al., The Assessment Report of the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services on Pollinators, Pollination and Food Production. Bonn, Germany: Secretariat of the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services, 2016.
28. Cost Estimates of Studies Required for Pesticide Registration, Washington, DC, USA: U.S. Environmental Protection Agency, May 13, 2024. [Online]. Available: https://www.epa.gov/pesticide-registration/cost-estimates-studies-required-pesticide-registration. Accessed: Apr. 25, 2025.
29. Ecological Effects Test Guidelines: Honey Bee Toxicity of Residues on Foliage (OCSPP 850.3030), Washington, DC, USA: U.S. Environmental Protection Agency, Jan. 2012. [Online]. Available: https://www.regulations.gov/document/EPA-HQ-OPPT-2009-0154-0017. Accessed: Apr. 25, 2025.
30. Xu P, Liu D, Diao J, Lu D, Zhou Z. Enantioselective acute toxicity and bioaccumulation of benalaxyl in earthworm (Eisenia fetida). J. Agric. Food Chem. 2009;57(18):8545-8549.
31. Huang L, Lu D, Diao J, Zhou Z. Enantioselective toxic effects and biodegradation of benalaxyl in Scenedesmus obliquus. Chemosphere. 2012;87(1):7-11.