Biomarkers of genotoxicity, immunotoxicity and oxidative stress on Caiman latirostris (Broad-snouted caiman) hatchlings exposed to pesticide formulations and mixtures widely used in agriculture

Pesticide formulations are currently considered as one of the main factors responsible for environmental contamination worldwide. Consequently, several wild species can be affected by the over use of pesticides related to agriculture activity. The aim of this study was to evaluate genotoxicity, immunotoxicity and oxidative stress on Caiman latirostris hatchlings exposed in vivo to sub-lethal concentrations of three insecticide formulations: Endosulfan (END)-, Cypermethrin (CYP)-, and Chlorpyrifos (CPF)- based formulations, two Glyphosate (GLY)- based herbicide formulations and two different ternary complex mixtures of them. C. latirostris hatchlings, 20 days old were exposed in plastics containers with pesticide solutions (by voluntary immersion) during two months, with a progressive decrease in pesticide concentrations through time, in order to simulate their degradation in water under natural conditions. After exposure, blood samples were taken to all animals for the analysis of genotoxicity (Comet assay, Micronucleus and other nuclear abnormalities) immunotoxicity (Total and differential white blood cell counts) and oxidative stress (Lipoperoxidation and the antioxidant enzymes Catalase and Superoxide dismutase). Results indicated that the five formulations tested, as well as the complex mixtures of them, induced genotoxicity, alteration in white blood cell counts and oxidative stress at environmentally relevant concentrations. Mixtures showed different interactions depending of the biomarker analyzed. This study constitutes an integral evaluation of the effect of five pesticide formulations and two possible complex ternary mixtures widely used in soybean crops in Argentina, on different endpoints on a native reptile species, C. latirostris.Fil: López González, Evelyn Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Romito, María Laura. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Latorre, María Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Siroski, Pablo Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentin

26th Annual Computational Neuroscience Meeting (CNS*2017): Part 3 - Meeting Abstracts - Antwerp, Belgium. 15–20 July 2017

This work was produced as part of the activities of FAPESP Research,\ud Disseminations and Innovation Center for Neuromathematics (grant\ud 2013/07699-0, S. Paulo Research Foundation). NLK is supported by a\ud FAPESP postdoctoral fellowship (grant 2016/03855-5). ACR is partially\ud supported by a CNPq fellowship (grant 306251/2014-0)

Total and differential white blood cell counts in Caiman latirostris after in ovo and in vivo exposure to insecticides

Agricultural activities associated mainly with soybean crops affect the natural environment and wildlife by habitat destruction and the extensive use of agrochemicals. The aim of this study was to evaluate immunotoxic effects of the insecticides cypermethrin (CYP) and endosulfan (END) in Caiman latirostris analyzing total blood cell count (TWBC) and differential white blood cell count (DWBC) after in ovo and in vivo exposure. Eggs (in ovo) and hatchlings (in vivo) from nests harvested in natural habitats were artificially incubated and reared under controlled conditions in the Proyecto Yacaré (Gob.Santa Fe/MUPCN) facilities. Exposure of embryos was performed by topication on the eggshell during the first stage of development. The treatments were distilled water (negative control; NC), ethanol (vehicle control; VC), four groups treated with different concentrations of CYP and four groups with END. In vivo exposure was performed by immersion; treatments were NC, VC, two groups exposed to CYP and two to END. After embryonic exposure to the insecticides, no differences were found in TWBC or DWBC among the neonates exposed to pesticides versus controls. In the in vivo scenario, similar results were obtained for TWBC, but DWBC data showed differences between NC hatchlings and CYP-1 hosts for heterophil, lymphocyte and monocyte levels, and between NC and END-1 hosts for lymphocyte and monocyte levels. Research on the effects of pesticide exposure on this species is of special interest not only to assess the impact on caiman populations, but also to further characterize the species as a potential sentinel of ecosystem health.Fil: Latorre, María Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias; ArgentinaFil: Romito, María L.. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias; ArgentinaFil: Larriera, Alejandro. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias; ArgentinaFil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Siroski, Pablo Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias; Argentin

Genotoxicity and oxidative stress in Caiman latirostris hatchlings exposed to pesticide formulations and their mixtures during incubation period

Agricultural expansion and the consequent use of pesticides lead to the loss and fragmentation of natural habitats of several wild species. Then, many species are inevitably exposed to a wide amount of pesticide formulations. Glyphosate (GLY)-based formulations are the most used herbicide, whereas two of the most employed insecticides are chlorpyrifos (CPF) and cypermethrin (CYP). The aim of this study was to evaluate genotoxicity, oxidative damage, and the modulation of antioxidants defenses in peripheral blood of Caiman latirostris after embryonic exposure to pesticide formulations and their mixtures. Pesticides concentrations employed were equivalent to those recommended in agricultural practices for application in soybean crops and a half of them: GLY: 2% and 1%; CYP: 0.12% and 0.06%; CPF: 0.8% and 0.4%. Two similar experiments (E1 and E2) were carried out in consecutive years, where C. latirostris eggs were exposed to pesticide formulations separately and in different mixtures through application on the incubation material. After hatching, blood samples were taken and genotoxicity and oxidative stress was evaluated through the micronucleus (MN) test, the modified comet assay, the lipid peroxidation (LPO) levels and the activities of catalase (CAT) and superoxide dismutase (SOD) antioxidant enzymes. The results indicated the presence of DNA damage, oxidation of purines and pyrimidines, and increased frequency of micronucleus (FMN) in the case of GLY, CYP, and CPF formulations exposure, as well as in all the mixtures tested, with respect to the control groups. Specifically, the results observed for the mixtures would indicate independent action or antagonism of the components for DNA damage and base oxidation (purines and pyrimidines) and a possible potentiation interaction for the FMN in two binary mixtures. However, there were not differences regarding lipid peroxidation, the activity of antioxidant enzymes and growth parameters. This study proved that the use of pesticide formulations at concentrations used in the field generate deleterious genetic effects on this species, then, exposure to them could threaten its survival and health status.Fil: Odetti, Lucia Magdalena. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: López González, Evelyn Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; ArgentinaFil: Romito, María Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; ArgentinaFil: Simoniello, Maria Fernanda. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentin

Biomarkers of Environmental Contamination in Reptile Species: The Effect of Pesticide Formulations on Broadsnouted Caiman Caiman latirostris (Crocodilia, Alligatoridae)

Pesticide formulations are currently considered as one of the main factors responsible for environmental contamination worldwide. Consequently, several wild species can be affected by the overuse of pesticides and fertilizers in agricultural activities. In recent years, we have been evaluating the effects of different pesticide formulations (glyphosate-, endosulfan-, cypermethrin- and chlorpyrifos-based formulations) and mixtures widely used in Argentina and other neighboring countries on broad snouted caiman (Caiman latirostris) as a sentinel reptile species for pesticide contamination. During this period, several instances of evaluation have been conducted in embryos and hatchlings, under controlled and semi-natural conditions. We used biomarkers of genotoxicity (micronucleus, other nuclear abnormalities, and the Comet assay) and oxidative stress (oxidation of lipids and DNA, and activity of the antioxidant enzymes catalase and superoxide dismutase), previously adapted by our group to be applied on blood, in order to avoid damage to the animals, as well as growth. All pesticides tested as well as the complex mixtures induced genotoxicity and oxidative stress through several biomarkers of different endpoints, in independent studies, but under these conditions, no synergistic effects were observed for the mixtures. The final aim is to continue with the integral in situ evaluation of environmentally exposed wild populations.Fil: Poletta, Gisela Laura. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: López González, Evelyn Cecilia. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Burella, Pamela Mariana. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romito, María Laura. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaFil: Siroski, Pablo Ariel. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; ArgentinaFil: Mudry, Marta Dolores. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentin