Abstract
The yellow-legged hornet, Vespa velutina subs. nigrithorax Buysson, 1905, is an invasive species in Europe, posing substantial ecological and economic threats. Its biology and behavior, marked by rapid reproduction and aggressive predation, endanger native insects. This has negative impacts on agriculture, biodiversity, and human safety. Effective control is therefore essential to prevent and minimize its spread and ecological impact. This study assessed the acute oral toxicity of four commercial insecticide formulations containing acetamiprid, spinosad, cypermethrin, and a mixture of natural pyrethrins as active ingredients on V. v. nigrithorax workers. Our findings indicate that acetamiprid and spinosad are the most promising compounds, with acetamiprid inducing rapid mortality at low doses, and spinosad causing both lethal and sublethal effects, potentially disrupting hornet behavior and colony viability. Given that oral exposure generally resulted in higher toxicity than contact exposure, pest control strategies should incorporate multiple exposure pathways. Further investigations are necessary to confirm efficacy under field conditions and to evaluate potential ecological risks to non-target species. Overall, this study provides critical data to improve V. v. nigrithorax control and supports the development of more effective and environmentally sustainable control strategies against this invasive species.
Similar content being viewed by others
Modeling abundance and risk impact of Vespa velutina nigrithorax (Hymenoptera: Vespidae) in Korea: application of a species abundance model
Arising amitraz and pyrethroids resistance mutations in the ectoparasitic Varroa destructor mite in Canada
Genetic and morphological variation of Vespa velutina nigrithorax which is an invasive species in a mountainous area
Data availability
The datasets generated during and/or analyzed during the current study are all available as Supplementary Material.
References
Grosso-Silva, J. M. & Maia, M. Vespa velutina lepeletier, 1836 (Hymenoptera, Vespidae), new species for Portugal. Arquivos Entomolóxicos 6, 53–54 (2012).
Walter, J. et al. First czech record of the Asian hornet (Vespa velutina nigrithorax) and a climatic prediction of its spread in the czech republic. BioI. Rec. 13, 607–620 (2024).
Rortais, A. et al. A new enemy of honeybees in europe: the Asian Hornet, Vespa velutina. Settele, J. Ed, atlas of biodiversity risks— from Europe to the Globe, from stories to maps. Pensoft, Sofia. 181 (2010).
Beggs, J. R. et al. Ecological effects and management of invasive alien Vespidae. Biocontrol 56, 505–526 (2011).
Pérez-de-Heredia, I., Darrouzet, E., Goldarazena, A., Romón, P. & Iturrondobeitia, J. C. Differentiating between gynes and workers in the invasive hornet Vespa velutina (Hymenoptera, Vespidae) in Europe. JHR 60, 119–133 (2017).
Rome, Q. et al. Caste differentiation and seasonal changes in Vespa velutina (Hym.: Vespidae) colonies in its introduced range. J. Appl. Entomol. 139, 771–782 (2015).
Franklin, D. N. et al. Invasion dynamics of Asian hornet, Vespa velutina (Hymenoptera: Vespidae): A case study of a commune in south-west France. Appl. Entomol. Zool. 52, 221–229 (2017).
Ruiz-Cristi, I., Berville, L. & Darrouzet, E. Characterizing thermal tolerance in the invasive yellow-legged hornet (Vespa velutina nigrithorax): the first step toward a green control method. PLoS ONE 15, e0239742 (2020).
Sánchez, O. & Arias, A. All that glitters is not gold: the other insects that fall into the Asian yellow-legged hornet Vespa velutina ‘specific’ traps. Biology 10, 448 (2021).
Barandika, J. F. et al. Efficacy of protein baits with fipronil to control Vespa velutina nigrithorax (lepeletier, 1836) in apiaries. Animals 13, 2075 (2023).
Rojas-Nossa, S. V., Mato, S., Feijoo, P., Lagoa, A. & Garrido, J. Comparison of effectiveness and selectiveness of baited traps for the capture of the invasive hornet Vespa velutina. Animals 14, 129 (2024).
Jeong, H. et al. Nutritional value of the larvae of the alien invasive wasp Vespa velutina nigrithorax and amino acid composition of the larval saliva. Foods 9, 885 (2020).
Mokkapati, J. S., Wnęk, A., Laskowski, R. & Bednarska, A. Acute oral and contact toxicity of three plantprotection products to adult solitaryBees osmia bicornis. Pol. J. Environ. Stud. 30, 4105–4113 (2021).
Souto, P. M. et al. Acute contact toxicity of insecticides for the chemical control of the invasive yellow-legged hornet Vespa velutina nigrithorax (Hymenoptera: Vespidae). PLOS ONE vol. 20 (2025).
Marques, A. F., Moreira, T. & Casaca, J. D. Manual de Boas Práticas na destruição de ninhos de Vespa velutina. (2018).
European Commission. EU Pesticides Database. https://ec.europa.eu/food/plant/pesticides/eu-pesticides-database/ppp/screen/home (2025).
Direção-Geral de Saúde of Portugal. List of authorized biocides. https://www.dgs.pt/saude-de-a-a-z1.aspx#saude-de-a-a-z/biocidas (2023).
Ulloa, K. A., Curkovic, S. T. & Araya, C. J. Toxicidad Oral de Seis insecticidas en larvas de Vespula (F) germanica en Laboratorio. Agricu. TÃcopyr 66, 133–140 (2006).
Carneiro, L. S. et al. Acute oral exposure to imidacloprid induces apoptosis and autophagy in the midgut of honey bee Apis mellifera workers. Sci. Total Environ. 815, 152847 (2022).
University of Hertfordshire. PPDB – Pesticide Properties DataBase. http://sitem.herts.ac.uk/aeru/ppdb/.
University of Hertfordshire. BPDB – Bio-Pesticides DataBase http://sitem.herts.ac.uk/aeru/bpdb/index.htm.
OECD. Test No. 213: Honeybees, Acute Oral Toxicity Test. (OECD,). https://doi.org/10.1787/9789264070165-en (1998).
OECD. Test No. 247: Bumblebee, Acute Oral Toxicity Test. (OECD). https://doi.org/10.1787/9789264284128-en (2017).
R Core Team. R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria https://www.R-project.org/ (2024).
Wickham, H. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag New York https://ggplot2.tidyverse.org (2016).
Tomizawa, M. & Casida, J. E. Neonicotinoid insecticide toxicology: mechanisms of selective action. Annu. Rev. Pharmacol. Toxicol. 45, 247–268 (2005).
Reid, R. J. et al. Assessing the acute toxicity of insecticides to the buff-tailed bumblebee (bombus terrestris audax). Pestic. Biochem. Physiol. 166, 104562 (2020).
Baines, D., Wilton, E., Pawluk, A., De Gorter, M. & Chomistek, N. Neonicotinoids act like endocrine disrupting chemicals in newly-emerged bees and winter bees. Sci. Rep. 7, 10979 (2017).
Bacci, L., Convertini, S. & Rossaro, B. A review of sulfoxaflor, a derivative of biological acting substances as a class of insecticides with a broad range of action against many insect pests. J Entomol Acarol Res. 50, (2018).
Besard, L., Mommaerts, V., Abdu-Alla, G. & Smagghe, G. Lethal and sublethal side-effect assessment supports a more benign profile of spinetoram compared with spinosad in the bumblebee Bombus terrestris. Pest Manag. Sci. 67, 541–547 (2011).
Breslin, W. J., Marty, M. S., Vedula, U., Liberacki, A. B. & Yano, B. L. Developmental toxicity of spinosad administered by gavage to CD® rats and New Zealand white rabbits. Food Chem. Toxicol. 38, 1103–1112 (2000).
Rabea, E. I., Nasr, H. M. & Badawy, M. E. I. Toxic effect and biochemical study of chlorfluazuron, oxymatrine, and spinosad on honey bees (apis mellifera). Arch. Environ. Contam Toxicol. 58, 722–732 (2010).
Biondi, A. et al. The non-target impact of spinosyns on beneficial arthropods. Pest Manag Sci 68, 1523–1536 (2012).
Soonwera, M., Moungthipmalai, T., Takawirapat, W. & Sittichok, S. Ovicidal and repellent activities of several plant essential oils against periplaneta americana L. and enhanced activities from their combined formulation. Sci. Rep. 12(1), 12070 (2022).
Passara, H. et al. Anise and fennel essential oils and their combination as natural and safe housefly repellents. Insects 16, 23 (2024).
Delabie, J., Bos, C., Fonta, C. & Masson, C. Toxic and repellent effects of cypermethrin on the honeybee: laboratory, glasshouse and field experiments. Pestic. Sci. 16, 409–415 (1985).
Thompson, H. M. Assessing the exposure and toxicity of pesticides to bumblebees ( Bombus sp.). Apidologie 32, 305–321 (2001).
Copping, L. G. & Duke, S. O. Natural products that have been used commercially as crop protection agents. Pest Manag. Sci. 63, 524–554 (2007).
Oliver, C. J., Softley, S., Williamson, S. M., Stevenson, P. C. & Wright, G. A. Pyrethroids and nectar toxins have subtle effects on the motor function, grooming and wing fanning behaviour of honeybees (apis mellifera). PLoS ONE 10, e0133733 (2015).
Hopwood, J. et al. How Neonicotinoids Can Kill Bees: The Science Behind the Role These Insecticides Play in Harming Bees. (The Xerces Society for Invertebrate Conservation, Portland, OR, 2016).
EFSA et al. Modification of the existing maximum residue levels for acetamiprid in various crops. EFS2 19, (2021).
Hall, H., Bencsik, M., Capela, N., Sousa, J. P. & De Graaf, D. C. Remote and automated detection of Asian hornets (Vespa velutina nigrithorax) at an apiary, using spectral features of their hovering flight sounds. Comput. Electron Agric. 235, 110307 (2025).
Farruggia, F. T. et al. A retrospective analysis of honey bee (Apis mellifera) pesticide toxicity data. PLoS ONE 17, e0265962 (2022).
Siddiqui, J. A. et al. Insights into insecticide-resistance mechanisms in invasive species: challenges and control strategies. Front. Physiol. 13, 1112278 (2023).
Darrouzet, E. Le Frelon Asiatique, Un Redoutable Prédateur – Le Connaître Pour Mieux Le Combattre. (Syndicat national d’apiculture, 2019).
Lioy, S. et al. Effectiveness and selectiveness of traps and baits for catching the invasive hornet Vespa velutina. Insects 11, 706 (2020).
Lee, C.-G. & Yu, S.-H. Exterminator for the nests of Vespa velutina nigrithorax using an unmanned aerial vehicle. Drones 7, 281 (2023).
Salgado, V. L. Studies on the mode of action of spinosad: insect symptoms and physiological correlates. Pestic. Biochem. Physiol. 60, 91–102 (1998).
Sparks, T. C. & Nauen, R. IRAC: mode of action classification and insecticide resistance management. Pestic. Biochem. Physiol. 121, 122–128 (2015).
Watson, G. B. Actions of insecticidal spinosyns on γ-aminobutyric acid responses from small-diameter cockroach neurons. Pestic. Biochem. Physiol. 71, 20–28 (2001).
Acknowledgements
We thank Marco Portocarrero and Bárbara Rodrigues from Associação Nativa, Joana Lopes and all the staff from Proteção Civil de Coimbra, and Henrique Silva from Câmara Municipal da Lousã for providing nest coordinates and for their invaluable support during fieldwork. We also thank Malaika Muschler, Giovanni Cilia, Leonhard Bürger, and Kimberley Lea Ring for their invaluable support during fieldwork. We also extend our gratitude to the two anonymous reviewers for their valuable contributions in reviewing the original manuscript of this work.
Funding
The study was carried out under the project “CONTROLVESPA—Development of strategies for the CONTROL of VESPA velutina invasion (PTDC/CTA-AMB/2123/2020)”, financed by FCT – Fundação para a Ciência e a Tecnologia, I.P. (DOI identifier https://doi.org/10.54499/PTDC/CTA-AMB/2123/2020). This study was also supported by the strategic plan of the Centre for Functional Ecology – Science for People and the Planet (CFE) (UIDB/04004/2020; https://doi.org/10.54499/UIDB/04004/2020) and Associate Laboratory TERRA (LA/P/0092/2020; https://doi.org/10.54499/LA/P/0092/2020).
Author information
Authors and Affiliations
Contributions
Conceptualization: Paula Souto, Artur Sarmento, Nuno Capela, Henrique M.V.S. Azevedo-Pereira; Data curation: Paula Souto, Artur Sarmento; Formal analysis: Paula Souto, Sara Leston; Funding acquisition: Henrique M.V.S. Azevedo-Pereira, José Paulo Sousa; Investigation: Paula Souto, Artur Sarmento, Nuno Capela, Soraia Santos, Aline Ronsani, Ana Luísa Tomaz, Sara Leston, Fernando Ramos; Methodology: Paula Souto, Artur Sarmento, Henrique M.V.S. Azevedo-Pereira, Nuno Capela, Soraia Santos, José Paulo Sousa; Project administration: José Paulo Sousa, Paula Souto; Resources: José Paulo Sousa, Fernando Ramos; Supervision: José Paulo Sousa, Paula Souto, Nuno Capela; Validation: Paula Souto, Artur Sarmento, Nuno Capela, José Paulo Sousa; Visualization: Paula Souto, Artur Sarmento, Nuno Capela; Writing – original draft: Paula Souto, Soraia Santos; Writing – review & editing: Paula Souto, Artur Sarmento, Soraia Santos, Aline Ronsani, Ana Luísa Tomaz, Nuno Capela, Henrique M.V.S. Azevedo-Pereira, Sara Leston, José Paulo Sousa. All authors read and approved the final version.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethics approval
Individuals of V. v. nigrithorax were collected from both private properties and public spaces. For collections on private properties, consent from the owners was obtained. In public areas, permission for collection was given by local authorities responsible for treating nests. Moreover, no special authorization is necessary to keep this species in laboratory conditions or to conduct insect animal testing within the EU. The fieldwork did not involve any endangered or protected species.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Supplementary Information 1
Supplementary Information 2
Supplementary Information 3
Supplementary Information 4
Supplementary Information 5
Supplementary Information 6
Supplementary Information 7
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Reprints and permissions
About this article
Cite this article
Malaquias Souto, P., Santos, S.S., Sarmento, A. et al. Assessing the oral toxicity of acetamiprid, spinosad, cypermethrin, and pyrethrins in the invasive hornet Vespa velutina nigrithorax.
Sci Rep (2025). https://doi.org/10.1038/s41598-025-31988-x
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-025-31988-x
Keywords
- Biological invasions
- Chemical control
- Ecotoxicology
- Hymenoptera
- Insecticide
- Vespidae
Source: Ecology - nature.com
