ABSTRACT
BACKGROUND AND AIM: The commercial formulations of the herbicide atrazine (cATZ) are widely employed in Brazilian agriculture, and, as a consequence, ATZ has been found at levels above that established by law in the river basins in Brazil. Although the toxicity of ATZ in fish is well documented, there are few studies on the recovery capacity after cATZ exposure. This work aimed to evaluate, using several biomarkers, the toxic effects of long-term exposure to the sublethal (3.57 mg/L) and nonlethal realistic (3.00 µg/L) cATZ concentrations followed by a recovery assay, in fingerlings of a Brazilian teleost, the Piaractus mesopotamicus (pacu). MATERIALS AND METHODS: Pacu fingerlings were housed in glass tanks and divided into the following experimental groups (two tanks/group): Exposure control = EC, recovery control = RC, the sublethal groups exposed to 3.57 mg/L of cATZ, (sublethal exposure group = SLE and sublethal recovery group = SLR) and the nonlethal groups treated with 3.00 µg/L of cATZ (nonlethal exposure group = NLE and nonlethal recovery group = NLR). The exposure assay was semi-static with a duration of 30 days and the recovery assay (after cATZ withdrawal) lasted 14 days. Several biomarkers were evaluated in fingerlings from all groups: The swimming behavior, the body weight gain, the micronucleus formation and nuclear alterations in erythrocytes, and the hepatic and renal histopathology analyzed by qualitative and semi-quantitative morphological methods (using light and electron microscopy). RESULTS: No significant difference in weight gain was observed among the groups after the exposure and recovery assays. The sublethal exposure induced impaired swimming movements, significant histopathological alterations, including necrosis in the liver and kidney, and a significant increase in the frequency of micronuclei in erythrocytes. The nonlethal exposure induced only subtle histopathological changes in the liver and kidney. After recovery assay, no genotoxic alteration was noted in pacu exposed to sublethal concentration, while the cATZ-induced kidney damage was partially reversed but not the hepatic injury. CONCLUSION: cATZ exhibits long-term toxic effects on pacu, even at relatively low concentrations, affecting mainly the liver and the kidney, and the effects of sublethal concentration are only partially reversed after cATZ withdrawal.
ABSTRACT
AIM: The aim of this work was to evaluate the sensitivity of Pacu fingerlings (Piaractus mesopotamicus) by measuring the effects of median lethal concentration (LC50) of atrazine (ATZ - 28.58 mg/L) after acute exposure (up to 96 h). MATERIALS AND METHODS: The fish were exposed to the LC50 of ATZ for 96 h (28.58 mg/L) in a static system. During the experiment, the fingerlings were randomly distributed in four glass tanks (50 L) containing dechlorinated water. Four glass tanks were for the control group, and four were for the ATZ-exposed group (n=4 per glass tank), given a total number of 16 animals tested per group. The genotoxicity was evaluated by micronucleus (MN) test in erythrocytes from peripheral blood. Qualitative and semi-quantitative histopathological analyses, and also ultrastructural study, were applied in liver and kidney samples. Finally, the content of heat shock protein (Hsp70) in the liver was evaluated by the western blotting method. RESULTS: The morphological alterations in the liver, which was associated with increased expression of Hsp70, included nuclear and cytoplasmic vacuolization, cytoplasmic hyaline inclusions, and necrosis. The kidney presented edema and tubular cell degeneration with cytoplasmic hyaline inclusion. The semi-quantitative histopathological analyses indicated that the liver was more sensitive than kidney to ATZ-induced damage. Ultrastructural analysis showed that ATZ caused membrane alterations in several organelles and increased the number of lysosomes in hepatocytes and kidney proximal tubular cells. Nevertheless, no significant difference was observed in MN frequency in erythrocytes comparing treated and control groups. CONCLUSION: These results indicated that ATZ-induced damage to the kidney and liver function, ATZ at the concentration tested did not induce a significant difference in MN frequency in Pacu erythrocytes comparing treated and control groups, and also that Pacu fingerlings may be a good bioindicator for testing freshwater contamination.
