Developmental toxicity, acute toxicity and mutagenicity testing in freshwater snails Biomphalaria glabrata (Mollusca: Gastropoda) exposed to chromium and water samples.

A protocol combining acute toxicity, developmental toxicity and mutagenicity analysis in freshwater snail Biomphalaria glabrata for application in ecotoxicological studies is described. For acute toxicity testing, LC50 and EC50 values were determined; dominant lethal mutations induction was the endpoint for mutagenicity analysis. Reference toxicant potassium dichromate (K2Cr2O7) was used to characterize B. glabrata sensitivity for toxicity and cyclophosphamide to mutagenicity testing purposes. Compared to other relevant freshwater species, B. glabrata showed high sensitivity: the lowest EC50 value was obtained with embryos at veliger stage (5.76mg/L). To assess the model applicability for environmental studies, influent and effluent water samples from a wastewater treatment plant were evaluated. Gastropod sensitivity was assessed in comparison to the standardized bioassay with Daphnia similis exposed to the same water samples. Sampling sites identified as toxic to daphnids were also detected by snails, showing a qualitatively similar sensitivity suggesting that B. glabrata is a suitable test species for freshwater monitoring. Holding procedures and protocols implemented for toxicity and developmental bioassays showed to be in compliance with international standards for intra-laboratory precision. Thereby, we are proposing this system for application in ecotoxicological studies.

Biomphalaria embryotoxicity test (BET): 60 years of research crossing boundaries for developing standard protocols.

Snail's embryotoxicity test is a suitable approach for toxicity assay of traditional and emerging pollutants, environmental risk assessment, as well as screening and development of new molluscicides. Among the snail species, Biomphalaria spp. has been indicated as a promising model system for developing standardized test protocols for assessing the chemical toxicity using early developmental stages. Thus, the current study aimed to review the data available in the scientific literature concerning the experimental approach, type of chemicals and the response of multiple biomarkers (survival, hatching rate, development delays, morphological and behavior changes) in snail embryos applied in toxicity tests. Revised data showed that the use of Biomphalaria embryos to assess chemical toxicity began in 1962. Snail's embryotoxicity test was applied mainly for analyzing the toxicity and development of new molluscicides, while its use in ecotoxicological studies is emerging. Biomphalaria glabrata was the main species analyzed. Embryos exposed to chemicals showed bioaccumulation, mortality, hatching inhibition, development delays, and morphological malformations, which were classified into four categories (hydropic, shell, cephalic and unspecified malformations). Besides, research gaps and recommendations for future research are indicated. Overall, the results showed that the Biomphalaria embryotoxicity test (BET) is a suitable tool for toxicity and health risk assessment.

Establishment of the comet assay in the freshwater snail Biomphalaria glabrata (Say, 1818).

The single cell gel electrophoresis or the comet assay was established in the freshwater snail Biomphalaria glabrata. For detecting DNA damage in circulating hemocytes, adult snails were irradiated with single doses of 2.5, 5, 10 and 20 Gy of (60)Co gamma radiation. Genotoxic effect of ionizing radiation was detected at all doses as a dose-related increase in DNA migration. Comet assay in B. glabrata demonstrated to be a simple, fast and reliable tool in the evaluation of genotoxic effects of environmental mutagens.