FP091 ATIVIDADE DA GLUTATIONA S-TRANSFERASE EM MACR

R. Santos, R. Santos, B.M. Roswag, Universidade Federal do Rio Grande FURG

Introdução As regiões alagadas do extremo sul do país possuem alta biodiversidade

de macrófitas aquáticas, portanto podem abrigar espécies ainda pouco estudadas que tenham expressiva atividade de GSTs. A cidade do Rio Grande estando inserida na planície costeira da América Latina contém a flora indicada para o estudo que se pretende. Com um papel fundamental no metabolismo de xenobióticos, as glutationa S-transferases (GSTs) são enzimas de detoxificação e podem ser induzidas ou inibidas frente à exposição a certos contaminantes. Enzimas de diferentes espécies vegetais são utilizadas como biotransformadoras de vários poluentes, diminuindo as concentrações destes no meio ambiente. Dessa forma, pretende-se relacionar a atividade das GSTs contidas nas raízes com a capacidade detoxificadora da planta. Espera-se que estas enzimas, presentes em inúmeros organismos, estejam contidas nos tecidos dos vegetais estudados. As espécies vegetais estudadas foram escolhidas de acordo com sua abundância no ecossistema local e seu biótipo de ocorrência. As estruturas vegetais analisadas foram raiz, caule e folha com base em um estudo que concluiu que essas partes possuem atividade enzimática mensurável. É possível que o hábito ecológico da planta tenha influência na sua capacidade de biotransformação, sobretudo em plantas flutuantes, uma vez que suas raízes encontram-se em contato direto com a água. Portanto as plantas foram aqui agrupadas em quatro biótipos, ou seja, em flutuantes, emergentes, submersas e anfíbias Dessa forma, os objetivos fora: (1) Avaliar a atividade da GST em raízes de quinze espécies de plantas macrófitas. (2) Relacionar a atividade enzimática com o biótipo de ocorrência das macrófitas. (3) Comparar raiz, caule e folha dos vegetais entre si quanto a atividade enzimática das GSTs, para assim eleger a estrutura que mais possui atividade.

FP092 Avaliação do potencial genotóxico e mutagênico de águas de piscicultura

L.D. Cavalcanti, L. Francisco, B. Crispim, R.B. Zanon, J.C. Mauad, M. Russo, Universidade Federal da Grande Dourados; a.b. grisolia, Universidade Federal da Grande Dourados / Faculdade de Ciências Biológicas e Ambientais; L. Viana Finoto, Universidade Federal da Grande Dourados

A aquicultura é uma atividade em expansão no Brasil quando comparada às demais atividades agropecuárias, os pesque-pagues representam uma das principais formas de comercialização do pescado no Mato Grosso do Sul, destacando-se especialmente na produção de bagres como o pintado e seus híbridos, devido ao seu filé com excelente qualidade organoléptica, características favoráveis para pesca esportiva e aceitação do público consumidor. Assim sendo, é importante que se avalie nos peixes propriedades mutagênicas e genotóxicas ocasionadas por contaminantes ambientais presentes na água, a fim de gerar informações para planos de controle da qualidade da água e da carne neste setor. O objetivo deste estudo foi avaliar a genotoxicidade e mutagenicidade em pintado amazônico provenientes de pesqueiro na cidade de Dourados-MS. As coletas ocorreram em dezembro de 2014 e março de 2015 em um pesqueiro na cidade de Dourados. Foram coletados 5 exemplares de pintado amazônico (Pseudoplatystoma sp. x Leiarius marmoratus) em cada período amostral. Após a captura, os peixes foram anestesiados o sangue foi coletado por punção caudal, em seguida extensões sanguíneas foram realizadas a fim de realizar a contagem de alterações metanucleares (genotoxicidade) e micronúcleo (mutagenicidade). Como resultado obtivemos que não houve diferença significativa entre as épocas para o teste de genotoxicidade e mutagenicidade. Apesar da baixa frequência de alterações metanucleares e micronúcleo, pode observar que no mês de dezembro a frequência de micronúcleos foi maior que no mês de março, já para as alterações metanucleares, a frequência foi maior em março. Com base nisso, pode-se inferir que de acordo com o

ensaio, a água não apresenta potencial genotóxico e mutagênico, contudo, faz-se necessário a realização de novos estudos a fim de avaliar de forma mais abrangente a qualidade da água.

FP093 Binary mixture effects by monoaromatic hydrocarbons on the tropical microcrustacean Mysidopsis juniae

M. Nascimento, Federal University of Sergipe; S. Loureiro, Universidade de Aveiro / Biology; M. Rubens dos Reis Souza, M. da Rosa Alexandre, Federal University of Sergipe / Chemistry; J. Nilin, Federal University of Ceará / Ecology

The monoaromatic hydrocarbons benzene, toluene and xylenes (BTX) are volatile compounds present in petroleum and its derivatives. Among the most produced chemicals worldwide, they are used in the chemical industry as raw material in the manufacture of plastics, rubbers, dyes, resins, pesticides and solvents. Contamination of aquatic environments by these substances may result in to adverse toxic effects on organisms of different biological levels. The present study aimed to evaluate the toxicity of BTX isolated and in binary mixtures using the acute toxicity test (96h) with the marine microcrustacean Mysidopsis juniae. To achieve that, organisms were exposed to BTX concentrations in order to identify the effects on survival, as well as to study the modes of action of the substances in mixture with the application of the theoretical models of Addition Concentration (AC) and Independent Action (AI). As a result of the experiments with isolated compounds it was possible to notice that xylene was the most toxic compound (LC50 16,10 ±2,4mg.L-1) followed by toluene

(LC50 38 ± 5,3mg.L-1) and benzene (LC50 78,03 ±2,9mg.L-1) respectively. When in

binary mixture deviations of the reference models for CA were observed in all experiments performed (dose ratio and antagonism). In the exposure of xylene and benzene the deviation that fit was the antagonism, this effect predicts that the mixture causes a less severe effect than the isolated substances. Where as the mixture of xylene and toluene was better explained by the deviation dose ratio where the toxicity was caused mainly by xylene. In the experiment combining toluene and benzene, the dose ratio pattern was observed as well, and the benzene was the main responsible for the toxicity. We concluded that BTX were toxic to mysids, both isolated and in binary mixtures, evidencing the importance of studies in this subject that can serve as support for evaluations and monitoring of the marine environments, considering the imminent risks of contamination in the environment and the wide scale of use of these compounds.

FP094 Bioaccumulation kinetics of copper in Ruditapes philippinarum exposed to increasing, continuous and pulsed exposure: implications for growth

l. santana, Laboratório de Avaliação de Contaminantes Orgânicos (LACOr) / LABOMAR-UFC; J. Blasco, Inst. Ciencias Marinas de Andalucia / ECOLOGY AND COASTAL MANAGEMENT; D.M. Abessa, Universidade Estadual Paulista - UNESP/CLP / Marine Biology and Coastal Management; O. Campana, CSIC Spanish National Research Council ICMAN

The effects of metal bioaccumulation on aquatic organisms depend on factors as magnitude, duration and frequency of exposure. The type of exposure affects the toxicokinetic processes in the organisms. In this study, 30-day toxicity tests were carried out on juveniles of the bivalve Ruditapes philippinarum exposed to increasing, continuous and pulsed exposure with copper-spiked estuarine sediments followed by 10-day recovery period. The aim of the study was to assess the interaction between the kinetics of subcellular copper partitioning and the growth response. Pre-exposure test concentrations after sediment spiking were 12.1 (low), 19.2 (medium) and 24.7 (high) mg Cu < 63µm g-1 _{OC, representing environmentally relevant concentrations for}

relatively low contaminated sediments. Copper concentration in unspiked sediment (2.1 mg Cu < 63µm g-1 _{OC) was adequate to prevent chronic effects to benthic}

invertebrates in marine environments. Bivalves were acclimated 7 days under controlled conditions. Tests were conducted exposing them to the sediments in a flow- through system during 40 days. Four treatments were set up: (1) control, kept in unspiked sediment; (2) increasing exposure, increasing copper concentrations during 10 days each; (3) continuous exposure, kept at the highest concentration for 30 days; and (4) pulsed exposure, alternating the highest concentration and the control every 10 days. Organisms were sampled at different times (T0 = initial time, T1 = 10 days, T2 = 20 days, T3 = 30 days and T4 = 40 days) and average wet weight was recorded to calculate growth rate. Tissue samples were treated to determine the net copper accumulation rates for total bioaccumulated copper, biologically detoxified (enzymes, metal-rich granules and metallothioneins-like proteins fractions) and biologically available (mitochondria, lysosomes, microsomes and heat-sensitive proteins fractions) metal pools. Results showed that the bivalves’ growth rate was inversely correlated to the bioaccumulation rate and that sublethal copper concentrations stimulated the detoxification mechanisms of the organism regardless the type of exposure. However, a large stimulatory effect on growth was observed during the recovery, associated to

significant negative accumulation rate values and dependent on the type of previous exposure. This suggested that, on individual and short-term basis, pulsed exposures have a more adverse effect compared to increased or continuous exposure scenarios.

FP095 Biochemical Effects of Water-Soluble Fraction of Oil (WSF) on the Amphipod Hyalella curvispina

J. del Brio, CITAAC, UNCo-CONICET, LIBIQUIMA, FAIN. / Departamento de Química; C. Montagna, CITAAC, UNCo-CONICET, LIBIQUIMA, FAIN, FACIAS / Quimica; A. Venturino, Universidad Nacional del Comahue / Quim. Biol. - Inst Biotecnología

The Province of Neuquén is located in the northern region of the Argentinean Patagonia, where the exploitation of energy resources is the most relevant economic activity. The oil and gas fields from this area produce 48% and 23% of Argentina’s total production; respectively. Therefore, this zone is exposed to oil pollution derived from drilling, production and transport of crude oil. During the last years, studies from the area have demonstrated the presence of hydrocarbons in some water courses and sediments. In addition, low concentrations of polyaromatic hydrocarbons (PAHs) such as naphthalene and pyrene were detected. Oil contamination causes a serious problem to aquatic life. Previous investigations of crude oil contamination have revealed that their toxicity is mainly due to the water-soluble fraction (WSF), rather than to the dispersed droplets of the fraction. Thus, the aim of this work was to evaluate the time- dependent effects of a sublethal dilution of WSF on the activity of detoxifying enzymes in adults of H. curvispina. Crude oil obtained from Chachahuen field, North Patagonia region and supplied by YPF Company was used to prepare the WSF. Amphipods were sampled from Los Barreales lake (LB) and Durán stream (DS), nearby the urban area of Neuquén City where the presence of hydrocarbons was previously determined. The enzymatic activity of glutathione-S-transferase (GST) and catalase (CAT) and reduced glutathione content (GSH), were assessed in both control and exposed (1:1600 dilution for 6, 24 and 48 hours) amphipods. Both LB and DS showed a significant increase (more than 100%) on GSH content after 6 hours of exposure compared to control groups. GST activity was significantly decreased after 24 and 48 hours of exposure in LB amphipods, while the ones from DS showed an increase on GST activity after 24 hours of exposure compared to control group. There was an increase of about 30% of CAT activity in LB organisms after 24 hours of exposure, while those from DS decreased CAT activity along whole exposure. In conclusion, control adults from the DS exhibited lower GSH content and GST activity than control adults from the LB, evidencing a differential response between the two populations in relation to the quality of the sampling site. Short-term exposure to WSF also produce oxidative stress in both populations in view of the changes observed in the components of antioxidant system.

FP096 Biochemical responses induced by coexposure of the nanomaterial titanium dioxide (rutile and anatase) and copper in Limnoperna fortunei.

S.M. Nunes, FURG Universidade Federal do Rio Grande / Programa de Pós Graduação em Ciências Fisiológicas (Instituto de Ciências Biológicas - ICB); M. Josende, FURG Universidade Federal do Rio Grande / Programa de Pós Graduação em Ciências Fisiológicas Instituto de Ciências Biológicas ICB; C. Ruas, M. Gelesky, FURG Universidade Federal do Rio Grande; J.M. Monserrat, Universidade Federal do Rio Grande -FURG / Programa de Pós Graduação em Ciências Fisiológicas Instituto de Ciências Biológicas ICB; J. Ventura-Lima, FURG Universidade Federal do Rio Grande / Programa de Pós Graduação em Ciências Fisiológicas Instituto de Ciências Biológicas ICB

The titanium dioxide nanomaterial (nTiO2) has been produced and incorporated into

various consumer products and commercial applications, due their catalytic, photocatalytic and ultraviolet (UV) light absorption properties. The wide applications of nTiO2 can lead to the release of this nanomaterial into the environment endangering

the organisms. In addition, nTiO2 has been used in environmental remediation because

of its ability to degrade or adsorb metals from water, and can coexist with metals such as Cu, for example. The nTiO2 naturally exists in the crystalline forms rutile and

anatase, which have different properties and can cause different effects in the organisms. This way, both nTiO2 and Cu can occur together in the environment, and

there is a need to study the potential combined effects of them. For this reason, this study aimed to evaluate if the coexposure to different crystalline forms of nTiO2 (rutile

and anatase) (1 mg/L) can influence in the toxicity and bioaccumulation of Cu (56 µg/L) in the golden mussel Limnoperna fortunei after 120 h of exposure. The following parameters were analyzed in the gills, digestive gland and adductor muscle: (1) quantification of Cu, (2) determination of the activities of glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) enzymes, and (3) assessment of the levels of lipid peroxidation (LPO). The results showed that coexpositions increased Cu accumulation in the three tissues independently of crystalline form. In the gills,

rutile and coexposure rutile + Cu increased GST activity and decreased SOD activity. In the digestive gland, anatase increased GST activity and decreased SOD activity. In the adductor muscle, rutile and coexposure rutile + Cu decreased the activity of GST and SOD. All treatments inhibited CAT activity in the gills and digestive gland, while in the adductor muscle the CAT activity was inhibited by rutile and coexposure rutile + Cu. Only rutile caused lipid damage in the gills. These results suggest that both crystalline forms exhibited toxicity and that coexposure of nTiO2 with Cu may be

harmful to L. fortunei, thus more attention in use and release of nTiO2 into

environment are needs to avoid effects in aquatic biota.

FP097 Biomarker responses in liver of zebrafish (Danio rerio) after acute exposure to sublethal concentrations of triclosan in water.

E.A. Sager, GECAP-Universidad Nacional de Lujan; P. Scarcia, INEDES UNLuCONICET / Departamento de Ciencias Básicas; D. Marino, Centro de Investigaciones del Medio Ambiente UNLP CONICET / Centro de Investigaciones del Medio Ambiente; A. Rossi, Instituto de Limnología INALI-CONICET; F.R. de la Torre, National University of Lujan / Basic Sciences

Triclosan (TC) is a widely used antibacterial/antifungal compound found in gels, toothpaste, deodorants, etc. The responses of hepatic biomarkers were evaluated in zebrafish (Danio rerio) after acute exposure to two sublethal concentrations of TC. Fish (391 ± 58 mg) were exposed to nominal concentrations of TC in water: 160 µg/L (TC1) and 340 µg/L (TC2) using a stock solution of TC dissolved in methanol, with a continuous flow system. Control fish remained in unchlorinated water (Ctrl) and water with the maximum concentration of methanol used (CtrlSv). Water samples were taken for determining TC exposure concentration by HPLC-MS. After 48 hours, fish were anesthetized, measured, weighed, sacrificed, and the liver was excised. Pooled samples were made up from 4 individuals, and 8 samples per treatment were processed (n=128). Enzymatic and non-enzymatic biomarkers were evaluated: activities of glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT), total antioxidant capacity against peroxyl radicals (ACAP) and lipid peroxidation levels as thiobarbituric acid reactive substance (TBARS). Statistical differences (p?0.05) between groups were determined using analysis of variance followed by Tukey`s test. No fish mortality was observed during assay period. There were no significant differences between Ctrl and CtrlSv in the different biomarkers tested. In addition no significant difference in SOD activity and ACAP level were observed. Acute exposure to TC promoted significant increase in TBARS levels by 20% in TC1 and TC2 with respect to Ctrl. However, an inhibition in CAT activity was detected in zebrafish exposed to TC1 (-24%) and TC2 (-34%) while GST activity decreased by 24% in both concentration relative to Ctrl. These results suggest that acute exposure of zebrafish to TC promoted an inhibitory response in liver at both, antioxidant activity (CAT) and biotransformation (GST). Furthermore, exposure to TC showed that lipid peroxidation in liver was promoted. Finally, both tested TC concentrations lead to similar biomarker responses after acute exposure.

FP098 Biomarkers in native fish present in the biobio river, central Chile, convergence effects of complex chemical mixtures

M. Quiroz Jara, Universidad de concepcion / Biomarcadores; M. Saavedra, Universidad de concepcion / Aquatic Systems, Faculty of Environmental Sciences; C. Melo, Universidad de concepcion / Biochemistry; J. Gavilán, Universidad de concepcion / Celular Biology, Faculty of Biological Science; R. Barra, Universidad de concepcion / Facultad de Ciencias Ambientales Eula

The Biobío River is the third most important basin in Chile with a 24.369km2 and supplies drinking water to about 970,000 inhabitants. Numerous companies use the water resource for their industrial processes, and as a consequence the elimination of effluents on their main channel, generating zones of punctual contamination (pulp and paper mill, oil refinery, among others), which due to the improvements in their technologies, have shown an increase in their productivity in recent years. Further, there are areas of diffuse pollution from the use of agricultural and forestry land, which together have affected the biota and water quality mainly in its lower third. To understand How the adjacent development alters the water quality and impacts the physiological state of native fish, in this work we identified sublethal responses evaluated using biomarkers strategy in native fish (EROD activity determined as pmol resorufin/mg prot./min), physiological indices (condition factor and hepatosomatic index), and environmental parameters (pH, temperature, conductivity, total dissolved solids). The obtained results exhibit an increase of the EROD activity evaluated in livers of the Percilia irwini species (n=106), presenting an induction gradient along the main course, with maximum values of 492±257 and 335±34 in females and males respectively, in the lower third of the river. These responses coincide to an increase in the environmental parameter values towards this zone and show a convergence effect of a complex mixture of chemical contaminants with respect to less impacted areas.

Consequently, the results of this work indicate a gradient of observable biological effects in the lower third of the Biobío River potentially attributable to the convergence of point and diffuse chemical contaminants in the study area.Mauricio Quiroz-Jara is scholarship CONICYT-Chile for PhD studies. Financial support for this study was provided by CRHIAM/Conicyt/Fondap 15130015.

FP099 Bisfenol A não altera atividade colinérgica de Oreochromis niloticus

J.d. Dias, L.M. Moraes, K.C. Cruz, A.O. Fernandes da Silva, Universidade Estadual do Norte do Paraná / CCHE

A tilápia-do-nilo (Oreochromis niloticus) é a espécie mais cultivada em piscicultura de todo o mundo O bisfenol A (BPA) é um monômero de plástico policarbonato que está presente em uma série de produtos de consumo e é constantemente liberado no ambiente, contaminando, assim, águas e solos. Estudos utilizando BPA resultaram em alterações enzimáticas em modelos de roedores, mas quase nada se conhece os efeitos do BPA sobre os peixes. A acetilcolinesterase (AChE) é uma enzima chave que encerra a neurotransmissão entre muitos tipos de células excitáveis. O objetivo deste trabalho foi analisar os efeitos agudos do Cd sobre a atividade da AChE muscular de tilápias-do-nilo. Os peixes foram aclimatados durante 7 dias. Foram separados em duplicata, os grupos controle (CTR, n=10) que receberam injeção de intraperitoneal (IP) de veículo apenas e o grupo experimental (EXP, n= 10) no qual inoculou-se via IP, 150 mg/kg de BPA. Os peixes foram mantidos em observação durante 30 dias alimentados diariamente com ração. Após o tratamenjto, os animais foram anestesiados e o músculo dorsal foi excisado, homogeneizado em tampão fosfato de potássio (0,1 M) para a determinação da atividade da AChE (nmol DTNB. min-1_.mg

de proteína-1_{) por espectrofotometria (415 nm), a concentração de proteínas totais}

foram quantificadas e o resultado expresso em médias (M) e erro padrão (SE). Realizou-se análise de variância paramétrica (ANOVA, P < 0,05). Não houve mortalidde de peixes ao longo do período experimental e os resultados demonstraram