Developmental toxicity, acute toxicity and mutagenicity testing in freshwater snails Biomphalaria glabrata (Mollusca: Gastropoda) exposed to chromium and water samples Lenita de Freitas Tallarico a,b,n , Sueli Ivone Borrely b , Natália Hamada b , Vanessa Siqueira Grazeffe a , Fernanda Pires Ohlweiler c , Kayo Okazaki b , Amanda Tosatte Granatelli a , Ivana Wuo Pereira d , Carlos Alberto de Bragança Pereira e , Eliana Nakano a a Laboratório de Parasitologia/Malacologia, Instituto Butantan, Avenida Vital Brasil,1500, CEP 05503-900, São Paulo, SP, Brazil b Instituto de Pesquisas Energéticas e Nucleares, IPEN-CNEN/SP, Av. Prof. Lineu Prestes, no 2242, Cidade Universitária, CEP 05508-000, São Paulo, SP, Brazil c Laboratório de Malacologia, Superintendência de Controle de Endemias, Rua Cardeal Arcoverde, no 2878, CEP 05408-003, São Paulo, SP, Brazil d Companhia de Saneamento Básico do Estado de São Paulo, R. Major Pinheiro Fróes, no 1.560, CEP 08680-000, Suzano, SP, Brazil e Departamento de Estatística, Instituto de Matemática e Estatística, Universidade de São Paulo,Rua do Matão, no 1010, CEP 05008-090, São Paulo, SP, Brazil article info Article history: Received 11 June 2014 Received in revised form 28 August 2014 Accepted 3 September 2014 Keywords: Biomphalaria glabrata Dominant lethal test Developmental toxicity Potassium dichromate Toxicity abstract 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, LC 50 and EC 50 values were determined; dominant lethal mutations induction was the endpoint for mutagenicity analysis. Reference toxicant potassium dichromate (K 2 Cr 2 O 7 ) 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 EC 50 value was obtained with embryos at veliger stage (5.76 mg/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. & 2014 Elsevier Inc. All rights reserved. 1. Introduction Acute and chronic toxicity tests have been mostly used in ecotoxicological protocols to evaluate the potential effects of environ- mental contaminants to natural populations (Cooney, 1995). However, with the need to assess the potential sublethal hazards to ecosystems of pollutants at low concentrations, environmental monitoring pro- grams have encouraged the analysis of effects on gametes, fertilization, reproduction and embryo-larval development (Llanos-Rivera et al., 2009). The effects on reproduction of many pollutants are unknown. Furthermore, among the myriad of chemicals reaching the environ- ment, some compounds classes can directly affect the reproductive potential through the induction of mutations in germ cells (Evenden and Depledge, 1997). Among aquatic organisms used in ecotoxicological studies, inverte- brates have been employed due to their importance in trophic chains and greater sensitivity response to chemical pollutants (Achiorno et al., 2010). Although mollusks are the second largest group in kingdom Animalia, they have not been considered in environmental risk assessment so far, mainly due to the lack of standardized protocols. In this sense, gastropods, the most abundant mollusks, have been successfully used as pollution indicators by different compound classes, such as, metals, pesticides an important group of emerging contaminants, called endocrine disruptors (Salice and Miller, 2003; Oliveira-Filho et al., 2005; Matthiessen, 2008; Ansaldo et al., 2009; Gagnaire et al., 2009; Giusti et al., 2013; Zounkova et al., 2014). Biomphalaria glabrata freshwater snails have been studied in many aspects because of their role as intermediate hosts of the trematode Schistosoma mansoni. Assays with B. glabrata (Say, 1818) Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/ecoenv Ecotoxicology and Environmental Safety http://dx.doi.org/10.1016/j.ecoenv.2014.09.005 0147-6513/& 2014 Elsevier Inc. All rights reserved. n Corresponding author at: Laboratório de Parasitologia/Malacologia, Instituto Butantan, Avenida Vital Brasil, 1500, CEP 05503-900, São Paulo, SP, Brazil. Fax: þ55 11 2627 9581. E-mail address: letallarico@gmail.com (L.d.F. Tallarico). Ecotoxicology and Environmental Safety 110 (2014) 208–215