Evaluation of the potential teratogenic and toxic effect of the herbicide 2,4-D (DMA® 806) in bullfrog embryos and tadpoles (Lithobates catesbeianus).

The objective of this work was to evaluate the potential teratogenic and toxic effect of the herbicide 2,4-D (DMA® 806) on bullfrog embryos and tadpoles (Lithobates catesbeianus). We used the FETAX (Frog Embryo Teratogenesis Assay Xenopus) assay for embryos, and for tadpoles, we used acute (96 h) and chronic (49 days) toxicity tests and evaluated aspects of healthiness, hematology, and histopathology. The LC50-144h (Median Lethal Concentration), EC50-144h (Median Effective Concentration), MCIG (Minimum Concentration to Inhibit Growth) and TI (Teratogenic Index) for embryos were 792 mg/L, 593 mg/L, 150 mg/L of 2,4-D (DMA) and 1.34, respectively. For tadpoles, the LC50-96h was 700 mg/L of 2,4-D (DMA) and chronic test indicated an inflammatory process and erythrocytosis (with possible polycythemia), with consequent reduction of the spleen. This demonstrates physiological stress probably due to dehydration, which can be proven by the gill tufts widening intercellular space and gill tuft fusions. We also found injuries to the kidneys and skin of the animals even in the lowest concentration tested. Our results indicated that this pesticide is minimally teratogenic and has a low toxicity on L. catesbeianus embryos and tadpoles, but it can inhibit embryo growth in concentrations lower than those tested in this study. We hypothesized that the herbicide 2,4-D (DMA® 806) may be a respiratory allergen for L. catesbeianus tadpoles and recommend precautionary measures for prolonged exposure of aquatic organisms to this pesticide.

Acute toxic and genotoxic effects of formalin in Danio rerio (zebrafish).

Formalin is a readily soluble chemical used as a sanitizing agent in the home and hospital. Formaldehyde solutions are routinely used in aquaculture for the prophylaxis and treatment of parasites and fungi, but the adverse effects of their application need to be further investigated. Danio rerio or zebrafish has characteristics favorable to its handling and breeding, and it is highly sensitive to various chemicals, being an ideal experimental model for this type of investigation. Thus, the objective of this study was to verify the toxic and genotoxic effects of formalin and to determine the lethal concentrations of this chemical to support its safe use in disinfection processes. Acute and chronic tests were performed using methods in accordance with international protocols. The genotoxic effect of formalin was evaluated with the micronucleus test using blood samples, which were collected at 96 and 192 h of exposure. The LC50-96h of formalin in D. rerio was 45.73 mg L-1, demonstrating its high resistance compared to other species. Regarding the genotoxic effect, the sublethal concentrations of formalin showed a positive correlation with micronuclei according to the increase in its concentration independent of the time of exposure. The incidence of micronuclei increased with concentration, and the addition of 1 mg L-1 formalin corresponded to an increase of 2.9% in the average number of micronuclei. In other words, formalin at even sublethal concentrations caused genotoxic effects in peripheral blood erythrocytes of D. rerio. Therefore, we recommend further studies and other tests involving this chemical for its use at environmentally safe concentrations.