Diclofenac and caffeine inhibit hepatic antioxidant enzymes in the freshwater fish Astyanax altiparanae (Teleostei: Characiformes)

Although concentrations of pharmaceutical compounds in aquatic ecosystems are low, they can cause toxic effects on organisms. The aim of this study was to evaluate the effects of diclofenac (DCF), a non-steroidal anti-inflammatory drug, and caffeine (CAF), a central nervous system stimulant, both alone or combined, in Astyanax altiparanae males under acute exposure (96 h), measuring neurotoxicity biomarkers, antioxidant response and damage at biochemical and cellular levels. DCF concentration in water, separated and combined, was 3.08 mg L−1 and that of CAF was 9.59 mg L−1. To assess neurotoxicity, brain and muscle acetylcholinesterase (AChE) activities were measured. To evaluate oxidative stress, the enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione S-transferase (GST), as well as lipoperoxidation (LPO), were analyzed in liver and gills. Activity of hepatic cyclooxygenase (COX) was also evaluated. Genotoxicity was assessed in blood using comet assay and micronucleus test, as well as nuclear abnormalities. DCF and CAF, alone or combined, had neither effect on AChE activity, nor in the activity of SOD, CAT, GPx and GST in gills. In liver, DCF inhibited SOD and GPx activity, CAF inhibited CAT activity, the mixture inhibited SOD and GST activity; although only fish exposed to CAF showed increased hepatic LPO. Under these experimental conditions, no effect on COX activity was observed, nor cytotoxic and genotoxic damage. The most pronounced effects were caused by the drugs separately, since both compounds altered the enzymes, but only CAF triggered LPO, showing more harmful effects.Fil: Muñoz Peñuela, Marcela. Universidade de Sao Paulo; BrasilFil: Lo Nostro, Fabiana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Dal'Olio Gomes, Aline. Universidade de Sao Paulo; BrasilFil: Tolussi, Carlos Eduardo. Universidade Anhembi Morumbi; BrasilFil: Branco, Giovana Souza. Universidade de Sao Paulo; BrasilFil: Pinheiro, João Paulo Silva. Universidade de Sao Paulo; BrasilFil: Godoi, Filipe Guilherme Andrade de. Universidade de Sao Paulo; BrasilFil: Moreira, Renata Guimarães. Universidade de Sao Paulo; Brasi

The role of ovarian steroids in reproductive plasticity in Hoplias malabaricus (Teleostei: Characiformes: Erythrinidae) in tropical reservoirs with different degrees of pollution

Chemicals in aquatic environments may lead to impairment of fish reproduction, but some species display plasticity levels to survive and reproduce in such localities. We investigated the reproductive cycle in a teleost species, Hoplias malabaricus, which inhabits a variety of environments with different degrees of pollution. Adult females were sampled at two locations in São Paulo State (Brazil), the reference Ponte Nova (PN) reservoir and the polluted Billings (BIL) reservoir. Metabolic and endocrine processes were analyzed throughout the annual cycle with special attention to ovarian histology, gonadosomatic index (GSI), fecundity, ovarian energetic substrates and plasma steroids analyses, in addition to physical and chemical analyses of the water. Pollutants were frequent in BIL but not in PN. The GSI, including the predominance of vitellogenic oocytes, was higher in spring and summer in both locations, but the oocyte recruitment dynamics was different. During winter, females from BIL presented vitellogenic oocytes and high levels of 11-ketotestosterone, which indicated precocity in the vitellogenic phase in relation to the females from PN. In animals from PN, high deposition of lipids occurred in the ovaries. However, plasma estradiol levels did not vary throughout the annual cycle. In animals from BIL, plasma estradiol levels peaked during the summer, but the ovarian lipid content remained unchanged throughout the year. The data suggest that the presence of pollutants cause various endocrine and metabolic responses and especially affects reproductive plasticity, which could explain why H. malabaricus survives under unfavorable environments.This work was supported by the São Paulo Research Foundation (FAPESP) (Research Grant: 12/50371-2 and a PhD scholarship: 2010/50555-0). The authors would also like to thank the fishermen (Evaldo Bizarrias and Milton Nunes de Santana) and the LAMEROA team for their help with sample collection and IB/USP for providing logistics and facilities for the collections

Influence of temperature in growth and physiological responses in Surubem dy Paraíba Steindachneridion parahybae (Siluriformes: Pimelodidae) in captivity

As mudanças climáticas devido às ações antrópicas tendem a alterar diversos fatores ambientais, entre eles a temperatura. Um recente prognóstico sobre a elevação da temperatura menciona que em uma escala otimista a temperatura na Terra se elevará em 1.8 graus C até 2100. Esta alteração pode causar diversas modificações nos processos fisiológicos, justificando a realização de estudos para avaliar os efeitos da temperatura na fisiologia dos animais. A Bacia do Paraíba do Sul há muito tempo, vem sofrendo com ações antrópicas, culminando no risco de extinção de espécies endêmicas, sendo este o caso do surubim do Paraíba, Steindachneridion parahybae. Com a possível elevação da temperatura ambiental o risco de extinção da espécie pode se tornar ainda mais eminente, sendo assim, o objetivo do presente trabalho é avaliar se a variação da temperatura da água, dentro dos valores previstos na escala otimista, de 1,8 graus C influencia o crescimento e as respostas fisiológicos dos juvenis de surubim do Paraíba. Foi estabelecido um modelo experimental no qual dois tanques foram cobertos com uma estufa de plástico (grupo Estufa) e outros dois tanques permaneceram sem cobertura (grupo Sem Estufa), com um número inicial de 100 peixes em cada tanque. Nos meses de setembro de 2009 (início), novembro de 2009, janeiro e março de 2010 foram realizadas biometrias em todos os animais, obtendo-se os dados morfométricos e ponderais, além de calcular os seguintes parâmetros zootécnicos: índice hepatossomático (IHS), fator de condição (K), crescimento relativo (CR), taxa de crescimento específico diária (CE), coeficiente de variação (CV), eficiência alimentar (EA) e sobrevivência (S). Amostras dos tecidos musculares e hepáticos, além do plasma foram coletadas em 5 animais de cada tanque experimental nas biometrias, sendo analisada a concentração de lipídeos, proteínas e carboidratos, perfil percentual de ácidos graxos da fração neutra (triglicérides) e polar (fosfolipídios). No plasma foi quantificada ainda a concentração de cortisol e glicose. Os animais do grupo Estufa, no qual a temperatura da água foi mantida cerca de 2 graus C acima do grupo Sem Estufa, apresentaram um maior incremento de massa corpórea em relação aos animais do grupo Sem Estufa, sendo o mesmo padrão observado para CR, CE e EA. Mesmo com um maior crescimento a concentração de lipídeos e proteínas tanto no músculo quanto no fígado foi igual entre os dois grupos, o mesmo ocorrendo com o glicogênio hepático. Por outro lado, o glicogênio muscular foi mais elevado na última coleta nos animais mantidos em temperaturas mais baixas. Na fração polar, os animais de ambos os grupos apresentaram uma respostas de elevação da porcentagem de ácidos graxos saturados e uma queda de polinsaturados, provavelmente relacionado à adaptação homeoviscosa, devido ao gradual aumento da temperatura. Na fração neutra, observou-se que os animais mantidos nas temperaturas mais elevadas utilizaram uma porcentagem maior de ácidos graxos polinsaturados para a obtenção de energia, o que em longo prazo pode ser prejudicial nos processos reprodutivos. Estes resultados sugerem que a elevação da temperatura pode ter ocasionado uma maior atividade enzimática e/ou uma maior eficiência destas enzimas e consequentemente maior crescimento e eficiência alimentar nos animais do grupo Estufa. No entanto, os resultados não permitem estabelecer se o aumento temperatura seria benéfico em todas as fases de desenvolvimento para o surubim do Paraíba.The climatic changes due to anthropogenic actions tend to alter several environment factors, among them, temperature. A recent prognostic about the temperature increase mentions that in an optimistic level the earth temperature will raise 1.8 degress C until 2100. This change may cause many modifications on the physiological processes, justifying the studies to assess the effects of the temperature in animals physiology. The Paraíba do Sul Basin, for a long time, has been suffering with the anthropogenic actions, culminating in the extinction risk of the endemic species. This is the case of surubim do Paraíba, Steindachneridion parahybae. With the possible rise in the environmental temperature, the risk of species extinction can be even more eminent, therefore, the aim of this study is to evaluate if the water temperature variation, within the values predicted in the optimistic level, i.e., 1.8 degrees C influences in the growth and in the physiological processes of surubim do Paraíba juveniles. It was established an experimental design in which two ponds were covered with a plastic greenhouse (Greenhouse group) and another two ponds were kept without the plastic coverage (No Greenhouse group), with an initial population of 100 animals in each tank. In September/2009 (beginning), November/2009, January/2010 and March/2010 the biometrical parameters were registered in all animals in order to obtain the morphometric and weight data, and in addition, to calculate the following parameters: hepatossomatic index (HSI), condition factor (K), specific growth rate (SGR), daily specific growth rate (DSGR), coefficient of variation (CV), feed efficiency (FE) and survival (S). The muscle and liver samples, and also the plasma were collected from 5 fish from each experimental tank in each sampling, and the tissue concentration of carbohydrates, proteins and lipids, fatty acids percentage profile in the neutral (triglycerides) and polar (phospholipids) lipids fractions were measured. The cortisol and glucose concentration were also quantified in the plasma. The Greenhouse group animals, where the water temperature was kept around 2 degrees C higher than the No Greenhouse group, showed an increased weight gain in relation to the animals of the No Greenhouse group, with the same result also found for SGR, DSGR and FE. Even with an increased growth, the proteins and lipids concentration both, in muscle and in the liver were unchanged between both groups, and the same occurred with the liver glycogen. However, the muscle glycogen was higher in the last sampling in the animals vi that were kept in the lower water temperature. In the polar fraction, both groups presented an increase in the percentage of saturate fatty acids and a decrease in the polyunsaturated percentage, probably related with a homeoviscous adaptation, due to the gradual increase in temperature. In the neutral fraction, the animals kept in higher water temperature utilized a higher percentage of polyunsaturated fatty acids as energy source, that, in long-term may be prejudicial for reproduction. These results suggest that the increasing temperature could result in a higher enzymatic activity and/or a higher efficiency of these enzymes and consequently, increased growth and feed efficiency in Greenhouse group animals. However, the results do not allow establishing whether the increasing temperature would be beneficial at all stages of development in the surubim do Paraíba

Anthropic influence of aquatic environment in the physiological processes associated with oocyte maturation in teleost fish

A ecotoxicologia emergiu como uma necessidade de avaliar e mitigar os impactos antrópicos no ambiente natural e nas comunidades viventes no local, utilizando uma gama de experimentos em campo e em laboratório. Dentre as diversas áreas estudadas a reprodução é um dos processos que podem ter seu desempenho diminuído devido à ação antrópica, sendo assim importante seu estudo na ecotoxicologia. O objetivo deste trabalho foi avaliar a influência do ambiente impactado nos processos fisiológicos associados à maturação oocitária em fêmeas de Astyanax fasciatus. Fêmeas de A. fasciatus foram coletadas entre 2012 e 2014 e machos (como indicadores da presença de compostos estrogênicos) entre 2013 a 2014, com duas coletas a cada estação do ano em dois reservatórios, Ponte Nova (referência) e Billings (impactado). Amostras de água e sedimento foram também coletadas nos mesmos pontos para análise dos parâmetros físicos e químicos. Estes resultados foram comparados com a resolução CONAMA/357 e calculado o coeficiente de risco. Para ambos os sexos foram analisadas a expressão gênica da vitelogenina e do receptor de estradiol, a concentração plasmática de vitelogenina e estradiol. Foram também realizadas as análises de fecundidade relativa e absoluta, histologia das gônadas, assim como o cálculo dos índices gonadossomático e hepatossomático, apenas nas fêmeas. O coeficiente de risco mostrou que o reservatório Billings apresenta um alto risco para os metais e dos compostos orgânicos. Foram observados valores elevados na concentração plasmática de estradiol e expressão de vitelogenina em machos deste reservatório, na primavera de 2013, assim como foram observados valores mais elevados de fecundidade relativa e absoluta nas fêmeas, no inverno e primavera deste mesmo ano, em comparação com os animais do reservatório referência (Ponte Nova). Esses resultados sugerem que no inverno e primavera de 2013, os poluentes como os aqui analisados e outros aqui não detectados, podem ter interferido na reprodução em A. fasciatus do reservatório Billings. No entanto, é necessário destacar que essas alterações podem, em dado momento, prejudicar o processo de maturação oocitária, sendo então importante monitorar e realizar mais estudos de análise de risco ambiental no reservatórioThe ecotoxicology has arisen as a need to evaluate and mitigate the anthropic impacts in wild environmental and in the local communities, using several bioassays and wild experiments. Among the several studied areas, the reproduction is a process that can have its performance reduced due to the anthropic impacts, showing the importance of studying these processes in ecotoxicologydy. The aim of this study was to evaluate the influence of the impaired environmental in the physiological processes associated with oocyte maturation of Astyanax fasciatus females. Astyanax fasciatus females were sampled between 2012 and 2014 and males (as indicators of the presence of estrogenic compounds) from 2013 to 2014, with two samplings at each season, in two reservoirs, Ponte Nova (reference) and Billings (impacted). Sediments and water were sampled in the same sites to evaluate the chemical and physical parameters. The results of the analysis were compared with the CONAMA/357 guidelines and the hazard risk was calculated. The gene expression of vitellogenin and estradiol receptor, vitellogenin and estradiol plasma level were performed in males and females. The relative and absolute fecundity, gonad histology, gonadosomatic and hepatosomatic index were evaluated only in females. The hazard risk at Billings reservoir was high for organic compounds and metals. Estradiol levels and vitellogenin gene expression were higher in males sampled in Billings in the spring/2013, as well as the relative and absolute fecundity, that were higher in the winter and spring of the same year, when compared with the animals from the reference site (Ponte Nova). These outcomes suggest that during winter and spring of 2013 the pollutants, as analyzed in this study, as well as others that were not detected, may have interfered in the reproductive process of A. fasciatus at Billings reservoir. However, it is necessary to observe that these alterations might, at a given time, impair the oocyte maturation process, being important to monitor and accomplish additional studies of the environmental risk assessment at Billings reservoi

The effects of stocking density in physiological parameters and growth of the endangered teleost species piabanha, Brycon insignis (Steindachner, 1877)

This study investigated the effects of stocking density on the growth and fatty acid (FA) of Brycon insignis metabolism. Fingerlings (360) were distributed into eight ponds at two stocking densities (105 and 210 g/m(3)). The analysis of growth showed that the condition factor (K) and the coefficient of variation (CV) for body mass were not affected by stocking density. However, final body mass and length, specific growth rate (SGR), and weight gain (WG) were higher in the low stocking density group, which also presented a higher feed efficiency (FE) and survival (S). By contrast, muscle protein levels were higher in the high stocking density group. The plasma and muscle lipid content were not affected by stocking density, but fish reared at lower stocking density presented higher lipid concentration in the liver, with no differences in hepatosomatic index values. Even with the differences observed in metabolic and growth parameters, plasma cortisol was not affected by stocking density. The FA profile in the muscle and liver neutral fraction were not affected by stocking density, but the FA in the polar fractions differed between the two stocking densities. In the liver, total polyunsaturated fatty acids (PUFA) and PUFA n - 3 increased in higher stocking density, mainly due to an increase in docosahexaenoic acid (DHA). In addition, PUFA n - 6 were also increased in the higher stocking density group, mainly due to an increase in arachidonic acid (AA) and docosadienoic acid (22:2n - 6). In the muscle polar fraction, the saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) decreased in the animals from the higher stocking density group, and this reduction was compensated by an increase in PUFA n - 3 and PUFA n - 6, mainly the FA with 20-22 carbons (20:4n - 6: 22:4:n - 6; 22:5n - 6, 22:5n - 3, and 22:6n - 3). A different profile was observed for the C18 PUFAs, mainly 18:2n - 6 and 18:4n - 6, which were higher in the lower density stocking group. The data suggest that when living in high stocking density, B. insignis differentially utilizes the hepatic lipids as energy source and remodels the membrane fatty acids, with higher amounts of DHA in the polar muscle fraction compensated for by a decrease in MUFA. The zootechnical and physiological indices reveal that the lower stocking density group achieve overall better performance. (C) 2010 Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP[2005/51302-0