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.

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.

Biomphalaria spp. 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.

Multiple adverse effects of textile effluents and reactive Red 239 dye to aquatic organisms

Textile dyeing consumes high volumes of water, generating proportional number of colored effluents which contain several hazardous chemical. These contaminants can implicate in significant changes in aquatic environmental, including several adverse effects to organisms in different trophic levels. The present study was developed to assess the ecotoxicological effects of textile effluent samples and reactive Red 239 dye (used in cotton dyeing) to aquatic organisms Vibrio fischeri bacteria, Daphnia similis crustacean, and Biomphalaria glabrata snail (adults and embryos). Chronic assays with lethal and sublethal effects for Daphnia similis were included and performed only for textile effluents samples. The mutagenicity was also evaluated with Salmonella/microsome assay (TA98, TA100, and YG1041 strains). V. fischeri bacteria was the most sensitive to reactive Red 239 dye (EC50 = 10.14 mg L−1) followed by mollusk embryos at all stages (EC50 = 116.41 to 124.14 mg L−1), D. similis (EC50= 389.42 mg L−1), and less sensitive to adult snails (LC50= 517.19 mg L−1). The textile effluent was toxic for all exposed organisms [E(L)C50 < 15%] and B. glabrata embryos showed different responses in the early stages of blastulae and gastrulae (EC50 = 7.60 and 7.08%) compared to advanced development stages trochophore and veliger (EC50 = 21.56 and 29.32%). Developmental and sublethal effects in B. glabrata embryos and D. similis were evidenced. In the chronic assay with effluent, the EC10/NOEC = 3% was obtained. Mutagenic effects were not detected for dye aqueous solutions neither for effluents samples. These data confirmed the importance of evaluating the effects in aquatic organisms from different trophic levels and reinforce the need for environmental aquatic protection.

The prospective use of Brazilian marine macroalgae in Schistosomiasis control

Schistosomiasis is a parasitic disease that affects more than 250 million people. The treatment is limited to praziquantel and the control of the intermediate host with the highly toxic molluscicidal niclosamide. Marine algae are a poorly explored and promising alternative that can provide lead compounds, and the use of multivariate analysis could contribute to quicker discovery. As part of our search for new natural compounds with which to control schistosomiasis, we screened 45 crude extracts obtained from 37 Brazilian seaweed species for their molluscicidal activity against Biomphalaria glabrata embryos and schistosomicidal activities against Schistosoma mansoni. Two sets of extracts were taxonomically grouped for metabolomic analysis. The extracts were analyzed by GC–MS, and the data were subjected to Pattern Hunter and Pearson correlation tests. Overall, 22 species (60%) showed activity in at least one of the two models. Multivariate analysis pointed towards 3 hits against B. glabrata veliger embryos in the Laurencia/Laurenciella set, 5 hits against B. glabrata blastula embryos, and 31 against S. mansoni in the Ochrophyta set. Preliminary annotations suggested some compounds such as triquinane alcohols, prenylated guaianes, dichotomanes, and xenianes. Despite the putative identification, this work presents potential candidates and can guide future isolation and identification

Bioassays with freshwater snails Biomphalaria sp.: from control of hosts in public health to alternative tools in ecotoxicology

Ecotoxicology is the science responsible for the study of the adverse effects of chemicals on ecosystems considering biotic and abiotic components. Several invertebrate groups have long been used to evaluate the aquatic toxicity of chemical compounds. Among these organisms, the microcrustaceans are the most recommended in Brazilian and international protocols (e.g. Daphnia sp. and Ceriodaphnia sp.). Until the beginning of the 1990s, the use of mollusks with ecotoxicological purposes was non-existent, except for the species tested as target of molluscicides in public health studies. Since the second half of this same decade the tests with mollusks have begun to be disseminated in several countries, valuing endemic species and especially the scarcity of test species in benthic habitats. In the early 2000s, with the disclosure of the harmful effects of pollutants with endocrine disrupting properties, gastropods have begun to be used not to evaluate lethal effects, but rather to observe physiological effects such as reproduction and embryonic development. Since then, assays with these approaches, especially with freshwater snails Lymnaea stagnalis and Biomphalaria sp., have been considered to be innovative and highly sensitive, often more than those achieved with traditional groups of test organisms in ecotoxicology (such as microcrustaceans and fishes)

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.

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.