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Leukocyte Profiles in Odontophrynus carvalhoi (Amphibia: Odontophrynidae) Tadpoles Exposed to Organophosphate Chlorpyrifos Pesticides

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Abstract

We analyzed the toxic effects of the organophosphate insecticide chlorpyrifos (Klorpan® 480 EC formulation) on the blood leukocyte profiles of Odontophrynus carvalhoi tadpoles, a neotropical species endemic to Brazil, exposed to four different concentrations of that insecticide (10, 100, 200, and 400 μg L−1) for 192 h. The tadpoles were anesthetized and their blood was collected by cardiac puncture, with subsequent cell smears and staining with May–Grunwald–Giemsa and Wright. The numbers and types of leukocytes were determined after different periods of exposure (24 h, 48 h, 96 h, and 192 h) by analyzing 100 leukocytes from each tadpole (under 1000× magnification). The tadpoles exposed to different concentrations of chlorpyrifos exhibited reductions in their numbers of lymphocytes, monocytes, and basophils, but increases in the numbers of neutrophils and eosinophils. As such, exposure to chlorpyrifos altered cell type proportions, which could alter the capacity of those tadpoles to respond to environmental stresses, make them more susceptible to infection by diverse pathogens, and thus compromise their chances of survival. That scenario would be even more serious for species having restricted distributions, as population declines would be more severe and impacting. Therefore, the use of chlorpyrifos constitutes a serious risk for anuran amphibians that occur in or near agricultural fields by contributing to immunosuppression and threatening their survival.

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Acknowledgments

The authors thank the Ministério do Meio Ambiente do Brasil for permission to undertake this research project (ICMBio license N° 27305-1); the Comitê de Ética no Uso de Animais da Universidade (CEUA-UEFS, protocol N° 02/2012); the anonymous reviewers for their commentaries and suggestions;

Funding

This research was financed by the National Council of Scientific and Technological Development—CNPq Edital MCT/CNPq N° 59/2009 Ecological Research of Long Duration Program PELD Chapada Diamantina and Research Support Foundation of the Sao Paulo State—FAPESP Project SISBIOTA Anura Tadpoles of the Mata Atlântica, Amazônia, Pantanal, Cerrado, and Zonas de Transição Morphological Characterization, Special Distribution, and Pattern of Diversity (FAPESP proc. 2010/52321-7 e CNPq proc. 563075/2010-4).

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  1. Laboratório de Zoologia, Instituto Multidisciplinar em Saúde, Campus Anísio Teixeira, Universidade Federal da Bahia, Rua Hormindos Barros, 58, Quadra17, Lote 58, Bairro Candeias, Vitória da Conquista, BA, 45029-094, Brazil

    Márcio Borba da Silva, Ricardo Evangelista Fraga, Patrícia Belini Nishiyama, Inaiara Sousa Siqueira da Silva, Natália Layane Badaró Costa, Luana Alzira Alves de Oliveira & Mariane Amorim Rocha

  2. Museu de Zoologia, Divisão de Anfíbios e Répteis, Universidade Estadual de Feira de Santana, Av. Transnordestina s/n, Bairro Novo Horizonte, Feira de Santana, 44036-900, BA, Brazil

    Flora Acuña Juncá

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  1. Márcio Borba da Silva
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  2. Ricardo Evangelista Fraga
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  3. Patrícia Belini Nishiyama
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Márcio B. Silva, Ricardo E. Fraga, Patrícia Belini Nishiyama, and Flora A. Juncá conceived and designed the experiments. Márcio B. Silva and Flora A. Juncá collected the amphibian specie. Márcio B. Silva, Ricardo E. Fraga, Inaiara S. S. Silva, Natália L. B. Costa, Luana Alzira Alves de Oliveira, and Mariane Amorim Rocha contributed to the manipulation of the experiment, the collection of blood, and the analysis of the samples. Márcio B. Silva, Ricardo E. Fraga, Patrícia Belini Nishiyama, and Flora A. Juncá contributed writing and revising the paper. All authors reviewed the manuscript and approved the submission.

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Correspondence to Márcio Borba da Silva.

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All procedures performed in this study involving animals were in accordance with the ASIH (2004) criteria and ethical standards of the animal ethics committee of the Universidade Estadual de Feira de Santana (CEUA-UEFS, N° 02/2012).

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Silva, M.B., Fraga, R.E., Nishiyama, P.B. et al. Leukocyte Profiles in Odontophrynus carvalhoi (Amphibia: Odontophrynidae) Tadpoles Exposed to Organophosphate Chlorpyrifos Pesticides. Water Air Soil Pollut 231, 372 (2020). https://doi.org/10.1007/s11270-020-04726-4

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