Mutagenic assessment of Lithobates catesbeianus tadpoles exposed to the 2,4-D herbicide in a simulated realistic scenario.

The aim of the current study is to assess possible erythrocyte mutagenic effects on Lithobates catesbeianus tadpoles exposed to water contaminated with 2,4-D. In order to do so, tadpoles were exposed to a predictive and environmentally relevant herbicide concentration (1.97 mg/L), which is likely to be found in lentic environments formed by superficial water runoffs in pasture areas where the herbicide was applied. The micronucleus test, as well as tests for other nuclear abnormalities, was conducted after 3, 5, and 9 days of exposure (d.e.). Changes in the biomass and mouth-cloaca length or interference in the larval development of the animals (in the three evaluated times) were not recorded. However, tadpoles exposed to 2,4-D showed the highest total number of nuclear abnormalities, as well as the highest frequency of binucleated erythrocytes and kidney-shaped nuclei (shortly after 3 d.e.). The micronucleus frequency was also higher in animals exposed to 2,4-D (in the 3rd, 5th, and 9th d.e.), as well as the frequency of binucleated cells (3rd, 5th, and 9th d.e.) presenting notched (9th d.e.) and blebbled (9th d.e.) nuclei in comparison to those of the control, after 5 and 9 days of exposure. Therefore, the current study is a pioneer in showing that 2,4-D has a mutagenic effect on L. catesbeianus tadpoles, even at low concentrations (environmentally relevant) and for a short period of time, a fact that may lead to direct losses in anuran populations living in areas adjacent to those subjected to 2,4-D herbicide application.