RESUMO

Endocrine disruptors, such as estrogen, are chemical substances with the potential to alter the hormonal balance of organisms. Their origin can be natural or artificial, and they can act at very low doses. The estrogen 17α-ethinylestradiol (EE2) is used worldwide as an oral contraceptive and is a potential contaminant in aquatic ecosystems. It is well documented that these environmental pollutants can act directly or indirectly on the reproductive system, impairing development and fertility. However, little is known about the alteration of the cell oxidative status induced by EE2. The main objective of this study was to evaluate the effect on the gill cells of adult zebrafish exposed in vivo to EE2, analyzing cell histology, DNA damage and the expression levels of genes encoding the main enzymes involved in oxidative stress pathways. The histological study showed that EE2 produces moderate to high damage to the gill tissue, an increase in gill cell DNA damage and the mRNA levels of the genes corresponding to the manganese superoxide dismutase (Mn-sod) and catalase (cat) after exposure to 5 ng/L EE2. The results indicate that EE2 causes tissue alterations, DNA damage and oxidative stress. EE2 produced important alterations in the gills, a fundamental organ for the survival of fish. There is a clear need for further research on the ecological consequences of EDCs on non-target organisms.

Assuntos

RESUMO

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.

Assuntos

RESUMO

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.

RESUMO

Superoxide dismutase (SOD) is an antioxidant enzyme that acts as a component of first-line defense system against reactive oxygen species (ROS). Copper/Zinc superoxide dismutase (Cu/Zn-SOD) is one of the isoforms of SOD enzyme and is sensitive to the exposure of different environmental factors, in different species and tissues. Caiman latirostris is one of the two crocodilian species living in Argentina and no information is available on the molecular and biochemical characteristics of the Cu/Zn-sod gene in this species. In the present work, we reported the presence of the Cu/Zn-sod gene in C. latirostris, the nucleotide and amino acid sequences, the modelled protein structure, evolutionary distance among species and tissue specific expression patterns. Cu/Zn-sod gene was 620 bp open reading frame in length and encoded 178 amino acids. The nucleotide sequences of C. latirostris shared high similarity with the Cu/Zn-sod genes of other crocodilian species, so it showed to be highly conserved. PCR analysis showed that Cu/Zn-sod gene was expressed in all the tissues examined (liver, gonads, spleen, heart, and whole blood), suggesting a constitutively expressed gene in these tissues. This study allows further investigation into the structure-activity relationship and the mechanism of action of Cu/Zn-SOD, besides exploring the functional breadth and possible alteration factors, including xenobiotics.

Assuntos

RESUMO

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.

Assuntos

RESUMO

The surface used for agricultural production in Argentina significantly increased in recent years, mainly due to the expansion of soybean crops. As a result, the use of agrochemicals increased too. Many natural populations of Caiman latirostris (broad-snouted caiman) are affected by habitat fragmentation and the constant exposure to pesticides. This exposure could produce Reactive Oxygen Species. The negative imbalance between ROS generation and the capacity of the biological systems to eliminate the reactive intermediaries or avoid the damage is called Oxidative Stress. The aim of this study was to evaluate oxidative damage and antioxidant defense in C. latirostris hatchlings after in ovo exposure to widely used pesticide formulations. Embryos were exposed by topical exposure on the eggshell, from the beginning of incubation period, to sub-lethal concentrations of two glyphosate formulations: PanzerGold® (PANZ) and Roundup® Full II (RU): 500, 750, 1000 µg/egg; to the endosulfan (END) formulation Galgofan® and the cypermethrin (CYP) formulation Atanor®: 1, 10, 100, and 1000 µg/ egg. Blood samples were taken to all animals immediately after hatching for the application and comparison of the following oxidative stress biomarkers between the exposed groups and their respective controls: lipoperoxidation through thiobarbituric acid reactive substances (TBARS), DNA base oxidation through the modified comet assay, and the activities of Catalase (CAT) and Superoxide dismutase (SOD) in erythrocytes. Our results showed lipoperoxidation in caiman exposed to END (10, 100, 1000 µg/egg), CYP (1, 10, 1000 µg/egg), RU (500, 1000 µg/egg) and PANZ (500, 1000 µg/egg), DNA base oxidation in those exposed to END (10, 100, 1000 µg/egg), CYP (1, 10 µg/egg) and PANZ (500, 750 µg/egg) as well as alteration in the activity of SOD in END 1 µg/egg and CYP (10, 1000 µg/egg). This study demonstrated the incidence of oxidative stress in animals exposed to pesticide formulations widely used in agricultural activity associated mainly with soybean crops, and add further information to that previously reported about pesticide effects in this native reptile species.

Assuntos