Implications of water hardness in ecotoxicological assessments for water quality regulatory purposes: a case study with the aquatic snail Biomphalaria glabrata (Say, 1818).

Water hardness is a property depending on the presence of alkaline earth metals, mainly calcium and magnesium. Among the strategies for water quality monitoring, ecotoxicological assays are performed to minimize impacts and classify water bodies. For these laboratory evaluations parameters are previously defined in the guidelines, including water hardness for both cultivation and testing medium. The present work was performed to evaluate the effects of different levels of water hardness on the survival and reproduction of the freshwater snail Biomphalaria glabrata and discuss the influence of natural water hardness on the results of ecotoxicological tests with these environmental samples. Comparing the groups it was possible to observe that those maintained in waters with least hardness had lower reproductive success, while the groups maintained in highest hardness showed better reproduction. These data show that waters with low hardness make the reproduction of the snail B. glabrata unfeasible, and this reveal a problem for ecotoxicity assays using natural water samples.

Effects of 17α-methyltestosterone on the reproduction of the freshwater snail Biomphalaria glabrata.

17-α-methyltestosterone (MT) is a synthetic hormone used in fish hatcheries to induce male monosex. Snails hold promise as possible test models to assess chemicals acting on the endocrine system. Biomphalaria glabrata is an aquatic gastropod mollusk (Pulmonata, Planorbidae) that can be easily maintained in aquaria, predisposing the species for use in ecotoxicological testing. This study evaluated the reproductive effects of MT on B. glabrata by examining histological changes and its reproductive performance. Ten snails per group were exposed for 4 weeks to different concentrations of MT (0.01, 0.1, and 1.0 mg/L). The total number of laid eggs, egg mass per group, size of type V oocytes, and production of spermatozoids were determined. Reproduction of B. glabrata was affected by MT. At the lowest concentration (0.01 mg/L), MT caused a statistically significant increase in the number of egg mass per snail compared with controls unexposed to MT. Histopathology analyses showed an increase in the sperm production at the higher MT concentrations of 0.1 and 1.0 mg/L. Chromatographic analyses of water samples showed that MT concentrations rapidly declined within a 96-h period. These results highlight the importance of giving more support to regulatory authorities, since MT is not registered for use on fish hatcheries in many countries around the world. Wastewater from fish farms discharged into aquatic ecosystems should be monitored for MT residues, since its presence could compromise the reproduction of other native snail species.

Implications of water hardness in ecotoxicological assessments for water quality regulatory purposes: a case study with the aquatic snail Biomphalaria glabrata (Say, 1818) / Implicações da dureza da água nas avaliações ecotoxicológicas com propósitos reguladores de qualidade da água: um estudo de caso com o caramujo aquático Biomphalaria glabrata

Water hardness is a property depending on the presence of alkaline earth metals, mainly calcium and magnesium. Among the strategies for water quality monitoring, ecotoxicological assays are performed to minimize impacts and classify water bodies. For these laboratory evaluations parameters are previously defined in the guidelines, including water hardness for both cultivation and testing medium. The present work was performed to evaluate the effects of different levels of water hardness on the survival and reproduction of the freshwater snail Biomphalaria glabrata and discuss the influence of natural water hardness on the results of ecotoxicological tests with these environmental samples. Comparing the groups it was possible to observe that those maintained in waters with least hardness had lower reproductive success, while the groups maintained in highest hardness showed better reproduction. These data show that waters with low hardness make the reproduction of the snail B. glabrata unfeasible, and this reveal a problem for ecotoxicity assays using natural water samples.

A dureza da água é uma propriedade dependente da presença de metais alcalino terrosos, principalmente cálcio e magnésio. Entre as estratégias para monitorar a qualidade da água ensaios ecotoxicológicos são realizados para minimizar impactos e classificar os corpos hídricos. Para essas avaliações em laboratório, parâmetros são previamente definidos nos protocolos, incluindo a dureza da água para cultivo e para a água de diluição. O presente trabalho foi realizado para avaliar os efeitos de diferentes níveis de dureza da água sobre a sobrevivência e a reprodução do caramujo de água doce Biomphalaria glabrata e discutir a influência da dureza de águas naturais nos resultados dos testes ecotoxicológicos com estas amostras ambientais. Comparando os grupos foi possível observar que aqueles mantidos em águas com menor dureza tiveram pior sucesso reprodutivo, enquanto os mantidos nas águas com maiores durezas tiveram melhor reprodução. Esses dados mostram que águas com baixas durezas tornam a reprodução do caramujo B. glabrata inviável, e esse fato revela-se como um problema para os ensaios ecotoxicológicos utilizando amostras de água naturais.

Genotoxic evaluation in Oreochromis niloticus (Fish: Characidae) of recombinant spore-crystal complexes Cry1Ia, Cry10Aa and Cry1Ba6 from Bacillus thuringiensis.

Bioinsecticides from Bacillus thuringiensis (Bt) are widely used around the world in biological control against larval stages of many insect species. Bt has been considered a biopesticide that is highly specific to different orders of insects, non-polluting and harmless to humans and other vertebrates, thus becoming a viable alternative for combating agricultural pests and insect vectors of diseases. The family of Bt δ-endotoxins are crystal-protein inclusions showing toxicity to insects' midgut, causing cell lysis leading to starvation, septicemia and death. The aim of this study is to evaluate the genotoxic potential of recombinant Bt spore-crystals expressing Cry1Ia, Cry10Aa and Cry1Ba6 on peripheral erythrocyte cells of Oreochromis niloticus, through comet assay, micronucleus (MN) test and nuclear abnormalities (NA) analysis. Fish (n = 10/group) were exposed for 96 h at 10(7) spores 30 l(-1), 10(8) spores 30 l(-1) or 10(9) spores 30 l(-1) of Bt spore-crystals. Cry1Ia showed a significant increase in comet cells at levels 1 and 2, but not at levels 3 and 4, so it was not mutagenic nor did it induce MN or NA. These three spore-crystals showed some fish toxicity at only the highest exposure level, which normally does not occur in the field.

Absence of tumor promoting activity of Euphorbia milii latex on the mouse back skin.

Euphorbia milii (Euphorbiaceae) is a decorative plant used in gardens and living fences. In China, it has also been employed in herbal remedies for hepatitis and abdominal edema. Since E. milii latex--lyophilized or in natura--proved to be a potent plant molluscicide, its toxicity to non-target organisms has been comprehensively studied. Concerns on a possible tumor promoting activity have discouraged its use as a locally-available alternative molluscicide in schistosomiasis control programs. Two in vitro assays (inhibition of metabolic cooperation in V79 cells and Epstein-Barr virus induction in Raji cells) had suggested that E. milii latex contained tumor-promoting substances. This study was undertaken to verify whether the latex acts as a tumor promoter in vivo as well. A single dose of the initiating agent DMBA (400 nmol) was applied on the back skin of male and female DBA/2 mice. Testing for tumor promoting activity began 10 days after initiation. Tetradecanoyl phorbol acetate (TPA) (5 nmol, positive control), lyophilized latex (20, 60 and 200 microg per mouse) or acetone (vehicle control) were applied on mouse back skin twice a week for 20 weeks. In TPA-treated mice, papillomas were firstly noted during the 11th week, and by the 17th week all animals exhibited skin tumors. No tumors developed in mice treated with the solvent alone and in those exposed to latex. Findings from the present study therefore indicated that E. milii crude latex does not act as a tumor promoting agent on the mouse back skin assay.

Toxicity of Euphorbia milii latex and niclosamide to snails and nontarget aquatic species.

The toxicity of Euphorbia milii molluscicidal latex and niclosamide (NCL) to target snails (Biomphalaria glabrata and Biomphalaria tenagophila) and nontarget aquatic organisms is evaluated. Planorbidae snails were killed by very low concentrations of lyophilized latex (48-h LC(50), mg/L: B. glabrata, 0.12; B. tenagophila, 0.09; Helisoma duryi, 0.10). Latex was less toxic (48-h LC(50) or EC(50), mg/L) to oligochaeta (Tubifex tubifex, 0.31), planktonic crustacea (Daphnia similis, 0.38; C. dubia, 1.07; Artemia sp., 0.93), and fishes (Danio rerio, 0.96; Poecilia reticulata, 1. 39), and considerably less toxic to Ampullariidae snails (Pomacea sp. , 10.55) and frog tadpoles (Rana catesbeiana, 7.50). Latex (up to 100 mg/L) was not toxic to bacteria (P. putida and V. fischeri), algae (Selenastrum capricornutum and Chlorella vulgaris), and mosquito larvae (Anopheles albitarsis, Aedes aegypti, Aedes fluviatilis). NCL was very toxic (48-h LC(50) or EC(50), mg/L) to Planorbidae snails (B. glabrata, 0.15, B. tenagophila, 0.13; H. duryi, 0.10), T. tubifex (0.11), crustacea (D. similis, 0.19; Ceriodaphnia dubia, 0.47; Artemia sp. 0.18), fishes (D. rerio, 0.25; P. reticulata, 0.29), R. catesbeiana (0.16), and Pomacea sp. (0.76). NCL was toxic to bacteria, algae (96-h IC(50), mg/L: S. capricornutum, 0.34; C. vulgaris, 1.23) and slightly toxic to mosquito larvae. In conclusion, E. milii latex, as compared with the reference molluscicide niclosamide, presents a higher degree of selectivity toward snails which are intermediate hosts of Schistosoma trematodes.

The influence of environmental factors on the molluscicidal activity of Euphorbia milii latex.

The present study was undertaken to investigate the influence of biotic (snail size and presence of food during the test) and abiotic factors (temperature, water hardness and concentration of organic materials) on the molluscicidal activity of Euphorbia milii latex. Bioassays were conducted with B. glabrata (10 snails per concentration) and snail lethality was evaluated after 24 hr and 48 hr of exposure to lyophilized latex solutions. Neither the degree of water hardness, nor the presence of food during the test affected latex-induced snail mortality. The snail size had a minor influence on E. milii-induced snail lethality. Newly-hatched (shell diameter < or = 1 mm) as well as young (3-8 mm) snails were slightly less susceptible than older (10-25 mm) mollusks. On the other hand, the molluscicidal effect of E. milii latex was modified by environmental factors such as temperature (i.e., LC50 and LC90 values were halved for every 10 degrees C rise in temperature) and concentration of organic materials in the water (i.e. the higher the concentration of organic matter, the higher the LC50 value). The efficacy of E. milii latex as a molluscicide can be modified by factors such as water temperature and concentration of organic materials, and to a lesser extent by snail size.