Baseline values of Micronuclei and Comet Assay in the lizard Tupinambis merianae (Teiidae, Squamata)

The Micronucleus test (MN) and Comet assay (CA) are currently the most widely used methods that allow the characterization of DNA damage induced by physical and chemical agents in wild species. The continuous expansion of the cultivated areas in Argentina, since the introduction of transgenic crops, mainly soy, in association with the increased use of pesticides, transformed deeply the natural environments where the lizard Tupinambis merianae occurs. Despite reptiles have shown to be excellent bioindicators of environmental contaminants, there is no record of genotoxicity studies in T. merianae. The aim of the present study was to adjust the MN test and CA protocols to be applied in erythrocytes of T. merianae, and determine the baseline values of DNA damage in this species. We used 20 adult lizards (10 males: 10 females) from Estación Zoológica Experimental “Granja La Esmeralda" (Santa Fe, Argentina). Peripheral blood samples were collected from all animals and the MN test and CA applied according to the protocols established for other reptilian species. We test critical parameters of CA protocol (cell density, unwinding and electrophoresis times) using increasing concentrations of H2O2 (10, 25 and 50 µM) as a known genotoxic agent to induce DNA damage. Based on this, we determined the most suitable conditions for the CA in this species: a cell density of 4 x 103 erythrocytes per slide, 10 min of unwinding and 15 min of electrophoresis at 0.90 V/cm approximately. The baseline frequency of micronuclei (BFMN= MN/1000 erythrocytescounted) determined for this species was 0.95 ± 0.27 and the basal damage index (BDI: calculated from 100 comet images classified in arbitrary units) = 103.85 ± 0.97. No differences were observed between sexes in the BFMN or BDI (p > 0.05), and no relation was found between baseline values and length or weight of the analyzed animals (p > 0.05). These results demonstrated the sensitivity of both biomarkers of genotoxicity to be applied in erythrocytes of this species, with baseline values comparable to those reported in other reptilian species. These results allow us to propose the tegu lizard for future in vivo studies to assess the genotoxicity of different agents, including those possibly affecting it in its natural geographic distribution.Fil: Schaumburg, Laura Gisela. 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: Poletta, Gisela Laura. 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: Siroski, Pablo Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); 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

Alterations in the Expression of Antioxidant Enzyme Genes in Response to Pesticide Exposure During Embryonic Development in the Native Reptile Species Caiman latirostris

The aim of this study was to quantify the expression levels of Catalase (cat) and copper, zinc Superoxide dismutase (Cu, Zn-sod) genes involved in the antioxidant response in Caiman latirostris (broad-snouted caiman) blood, after embryonic exposure to the formulations cypermethrin (CYP), chlorpyrifos (CPF), glyphosate (GLY), and their binary and ternary mixtures. Experimental groups were: negative control (NC-distilled water), vehicle control (VC-ethanol), GLY-2%, CYP- 0.12%, CPF- 0.8%, a ternary mixture of them (TM), and three binary mixtures. The applications were made on the nest material in contact with the eggs at the beginning of the incubation period. After hatching, RNA was isolated from blood and expression levels analyzed through qPCR. The results showed downregulation in the expression of sod and cat genes in the three binary mixtures studied, compared to the controls. In addition, we found a possible antagonistic effect between different pesticides in the TM on the expression of both genes.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. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Simoniello, M. F.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; ArgentinaFil: Poletta, Gisela Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

Effects of In Vivo Exposure to Roundup® on Immune System of Caiman latirostris.

The present study aimed to evaluate the effect of Roundup® (RU, glyphosate-based formulation) on some parameters of the immune system and growth of Caiman latirostris. Seventy-two caimans (20-day-old) from Proyecto Yacaré (Gob. Santa Fe/MUPCN) were used. Two groups were exposed for 2 months to different concentrations of RU (11 or 21 mg/L; taking into account the concentration recommended for its application in the field), while one group was maintained as control. The RU concentration was progressively decreased through the exposure period to simulate glyphosate degradation in water. Animals were measured and weighed at the beginning and end of the experiment, and blood samples taken after exposure to determine total and differential white blood cell (WBC) counts as well as total protein concentration (TPC), and for performing protein electrophoresis. The results showed that, compared against control hosts, there was a decrease in WBC counts, a higher percentage of heterophils, a higher TPC (with a low percentage of F2 protein fraction), and a negative effect on growth in the young caimans exposed to RU. These results demonstrate that in vivo exposure to RU induced alterations in the selected immune parameters, plasma proteins, and growth of caimans, thereby providing relevant information about the effects of this type of pesticide in this important species in the Argentinian wetlands.Fil: Latorre, María Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); 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: Larriera, Alejandro. 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; Argentina. 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 Ciencias Veterinarias; ArgentinaFil: Poletta, Gisela Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. 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; Argentin

Prenatal risk factors for selected congenital anomalies development: A case-control pilot study in postpartum women from Argentina

Introduction: Congenital anomalies (CAs) are abnormalities in intrauterine development and are one of the main causes of burden disease, especially in low and middle-income countries. Many environmental and maternal risk factors could lead to these pathologies. The aim of this study was to identify different types of CAs, determine their frequency and identify their possible association with maternal sociodemographic and lifestyle risk factors among patients from Santa Fe Province, Argentina. Material and methods: An observational cases and control pilot study was conducted in 2018 and 2019 on 280 postpartum mothers of newborn babies with any CA (cases, n = 64) and without a CA (controls, n = 216) attending public hospitals. A face-to-face questionnaire was completed, and clinical histories were required to obtain information on maternal sociodemographic factors, gynecological events, health state, lifestyle habits and child diagnoses. Results: Polymalphormations and neural tube defects were the most frequent CA observed. Differences between cases and control groups, as well as between places of residence, were given by gestational age, type of delivery and newborn birth weight, all of which are preventable variables. A slight association between maternal diabetes and the occurrence of cases was found. Discussion: Maternal place of origin is a factor of inequity in terms of gynecology variables which describes a deeper background in sanitary reality from Santa Fe Province that would have a large impact on future adults born preterm. The results highlight the necessity of generating both sanitary tools for maternal-child health policies and environmental evaluations, which remains a permanent challenge of the Argentine public health system.Fil: Colussi, Carlina Leila. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; ArgentinaFil: Racigh, Nora. Gobierno de la Provincia de Santa Fe. Hospital de Niños Doctor Orlando Alassia.; ArgentinaFil: Poletta, Gisela Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Simoniello, Maria Fernanda. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentin

Basal frequency of micronuclei and hematological parameters in the Side-necked Turtle, Phrynops hilarii (Duméril & Bibron, 1835)

The present study aimed to evaluate basaleline frequency of micronuclei (MN) and reference values for hematological values parameters in adult Phrynops hilarii in order to propose this aquatic turtle, broadly distributed in our region, as a biological monitor for future studies of environmental pollution assessment. Thirty-two adult turtles from a semi-natural environment located at the Zoological Experimental Station (Santa Fe, Argentina) were used. Blood samples were taken and the following parameters were determined: basaleline frequency of MN (BFMN), total red blood cells (RBC) count, hematocrit (Ht), hemoglobin (Hb), total and differential white blood cells (WBC). The BFMN determined for the species was 3.56 ± 1.39, while hematological parameters showed the following reference values: 0.937 ± 0.12 x106 RBC/µl, 27062.50 ± 4565.43 WBC/mm3, hematocrit 18 ± 1.81% and Hb concentration 4.80 ± 0.45 g/dl. Differential WBC counts were: 76 ± 2.90% for lymphocytes, 20.12 ± 2.56% for heterophils, 1.5 ± 0.19% for monocytes, and 2.12 ± 0.61% for eosinophils, while no basophils were observed in any of the samples analyzed. No differences were observed between males and females in any of the variables analyzed. Data provided in this work could be useful as reference values for future studies of natural regions where P. hilarii occurs, employing this species as a sentinel organism for genotoxic and immunotoxic evaluation of environmental pollutants.Fil: Latorre, María Agustina. 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; 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. Centro Científico Tecnológico Conicet - Santa Fe; 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: Poletta, Gisela Laura. 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); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris

The quantitative real-time polymerase chain reaction (qPCR) has been one of the most promising approaches to perform rapid and accurate quantification of DNA in various biological systems. The aim of this study was to standardized the qPCR technique for the analysis of important genes involved in the main routes of antioxidant defense against reactive oxygen species (catalase: cat and superoxide dismutase: sod) and evaluate the stability of different reference genes in blood of Caiman latirostris hatchlings. The stability of the reference genes, β-actin, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and ribosomal protein L8 (rpl8) was determined using the comparative ΔCt, NormFinder, geNorm, BestKeeper and RefFinder. Then, cat and sod genes were normalized with each reference gene and their mRNA abundances were determined through the qPCR. Stability of genes was ranked through the different methods in the following order: β-actin, rpl8 and gapdh, under normal physiological conditions. The results reveal that cat and sod genes present a similar relative mRNA abundance with β-actin and rpl8. This is the first report of the analysis of antioxidant mRNA as potential biomarkers of oxidative stress in blood for all crocodilians species. Besides, we determined the stability of different reference genes that can be used for normalization of mRNA abundance patterns in blood of C. latirostris, without the need to sacrifice the animals.Fil: Odetti, Lucia Magdalena. 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; ArgentinaFil: Paravani, Enrique Valentin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; ArgentinaFil: Simoniello, Maria Fernanda. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; 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

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

Efectos de la radiación ultravioleta natural y artificial (UVA/ UVB) sobre la concentración plasmática de calcio y fósforo y el crecimiento en crías de Caiman latirostris

La radiación ultravioleta interviene en la síntesis de vitamina D3 , indispensable para metabolizar calcio y fósforo. Naturalmente, la cantidad y calidad de radiación recibida por los individuos depende de numerosos factores. En condiciones artificiales es posible manipular la radiación e inferir los efectos producidos por dicha exposición sobre el crecimiento y la concentración plasmática de calcio y fósforo en diferentes organismos. En este trabajo se utilizaron 96 crías de C. latirostris de 4 meses de edad, provenientes de nidos cosechados en la naturaleza. Los tratamientos fueron: oscuridad total, 8 y 16 h de radiación ultravioleta artificial (A-B), y fotoperíodo natural (FN), durante 90 días. Se evaluó crecimiento (peso, longitud total y hocico-cloaca) y concentración de calcio y fósforo en plasma. No se observó efecto de los tratamientos sobre las concentraciones de calcio pero si sobre las de fósforo, manifestándose un descenso significativo en el tratamiento de 16 h (p< 0,05). Podemos suponer que la exposición a la que se sometió a las crías no fue suficiente para producir un comportamiento generalizado del metabolismo fosfocálcico. Aquellos animales expuestos al FN presentaron el mayor crecimiento (p< 0,05). Estos resultados indicarían que la exposición natural seria la óptima para el crecimiento de crías de C. latirostris.Fil: Fernández, L.. 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. 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; ArgentinaFil: Imhof, Alba. 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; Argentin

As ferramentas moleculares aplicadas na conservação dos crocodilianos

Nos últimos tempos, estudos de genética cresceram rapidamente graças ao avanço da tecnologia e das ferramentas moleculares, e os resultados obtidos nas análises genéticas se tornaram importantes ferramentas para a conservação e manejo, permitindo aprofundamento sobre a estrutura populacional, padrões de dispersão, fluxo gênico e hibridização interespecífica, dentre outros parâmetros. Os crocodilianos são um dos grupos de vertebrados mais fascinantes. Além de suas características intrínsecas (i.e., longevidade), representam um grupo bem-sucedido de organismos que sofreram poucas alterações ao longo de milhões de anos, sendo estes animais importante fonte de informação para quem pesquisa os avanços do ponto de vista molecular. A capacidade de resistir ao ataque de microrganismos poderia ser um dos motivos de seu sucesso e longevidade e, por isso, a descoberta dos componentes moleculares envolvidos permitiria um conhecimento aprofundado desses fatos. Neste aspecto, o entendimento dos processos evolutivos frequentemente requer a análise de marcadores polimórficos. Além disso, o impacto biológico resultante da interação com componentes sensíveis à contaminação em diferentes espécies de jacarés pode se manifestar nos níveis genético, imunológico e bioquímico, exercendo um efeito marcante de longo prazo nas populações. Marcadores moleculares, imunológicos e bioquímicos constituem uma bateria de ferramentas complementares altamente explicativas na avaliação dos efeitos nos crocodilianos causados pelo uso massivo dos pesticidas, permitindo obter mais informações sobre o significado biológico específico e o impacto real das alterações produzidas por defensivos agrícolas, gerando ferramentas preditivas para a proteção ambiental dessas e de outras espécies silvestres de nossa região. Especificamente, em toda a sua área de distribuição, um grande número de espécimes de C. latirostris foram caçados ilegalmente, razão pela qual foi necessário estabelecer diferentes estratégias para evitar sua extinção. Em conclusão, os recursos genéticos têm permitido abordar o estudo dos crocodilianos do ponto de vista populacional, ecológico, ecossistêmico e produtivo.Lately, genetics studies advanced quickly using molecular tools and the technology. Besides their intrinsic characteristics like longevity, habitats, and some other interesting aspects, crocodilians are one of the most fascinating groups within vertebrates. They represent a successful group of organisms that have suffered little alteration over millions of years, thus these animals are an important source of information for those researching advances from a molecular point of view. The ability to resist the attack of microorganisms could be one of the reasons for their success and longevity so the discovery of the molecular components involved would allow a thorough understanding of these facts. In addition, the information acquired from genetic analyses is important for conservation and management and allow to depth on population structure, patterns of dispersal and gene flow, interspecific hybridization, among other parameters, as to understand evolutionary processes require the analysis of polymorphic markers. Also, the biological impact resulting from the interaction with contamination-sensitive components in caiman species can be manifested at the genetic, immunological, and biochemical levels, exerting a marked long-term effect on populations. Genetic, immunological and biochemical molecular markers constitute a battery of complementary tools that are highly explanatory in the evaluation of the effects caused by mass use pesticides on crocodilians and will allow us to obtain more information regarding the specific biological significance and real impact of the alterations produced by pesticides on these species, providing predictive tools for the environmental protection of these and other wild species in our region.Fil: 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: Amavet, Patricia Susana. 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; 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 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; ArgentinaFil: Bassetti, Luís Antonio Bochetti. Universidade de Sao Paulo; Brasi

A comprehensive approach using multiple biomarkers to detect damage induced by pesticides in broad-snouted caiman (Caiman latirostris) under ex-situ conditions

Caiman latirostris is one of the two species of the order Crocodylia that inhabit Argentina and is considered a species of vital ecological and economic importance in the north-east of Argentina. In this region, pesticides are the most common contaminants in natural environments and wild caiman populations are subject to this contamination constantly. The aim of this study was to evaluate the effects the main pesticides used in the region: glyphosate (GLY), cypermethrin (CYP) and chlorpyrifos (CPF) -based formulations, as well as the mixture of them, on C. latirostris juveniles under semi-controlled condition of exposure (ex-situ) during 75 days. One hundred yearling caimans (10-month-old) were equally distributed into five experimental groups (20 animals per group): a negative control (NC -tap water), GLY 2% (Roundup® Full II formulation -RU), CYP 0.12% (Atanor® formulation), CPF 0.8% (Lorsban® formulation), and a mixture of the three pesticides (Mx3: GLY 2% + CYP 0.12% + CPF 0.8%). We applied early warning biomarkers to detect damage induced by these chemicals in peripheral blood: activity of the antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD), analysis of lipid peroxidation (LPO) by the thiobarbituric acid reactive substances (TBARS), DNA damage and specific base oxidation through the standard and modified comet assay (CA), chromosome damage by micronucleus (MN) test and other nuclear abnormalities (NAs), hematological and growth parameters. Results showed a statistically significant increase in MN and NAs frequency, DNA damage, with an important contribution of base oxidation for all exposed groups compared to the NC. Total white blood cells count (TWBCC), and growth parameters showed effects mainly at the Mx3. The principal component analysis (PCA) demonstrated more sensitivity for biomarkers associated to genetic damage, including base oxidation to DNA than LPO, antioxidant enzyme modulation, immunotoxicity or growth parameters, to detect pesticides effects, applied under conditions similar to that found in natural environments.Fil: 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. 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: Odetti, Lucia Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Latorre, María Agustina. 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: Ávila, Olga B.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Contini, Liliana Ester. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; 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: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentin