56 Áreas Marinhas Protegidas do Litoral Paulista sob Ameaça do TBT
F.B. Machado, Unifesp Universidade Federal de São Paulo; G.T. Sousa, I.B. de Castro, Universidade Federal de São Paulo / Instituto do Mar
Áreas Marinhas Protegidas (AMP) são fundamentais para proteção e preservação dos ecossistemas costeiros e oceânicos tendo como finalidade proteger zonas estratégicas de ameaças ambientais incluindo a contaminação por substancias químicas perigosas. Entre os químicos perigosos que podem afetar a vida marinha, o Tributilestanho (TBT), utilizado por muitos anos como ingrediente de tintas anti-incrustantes, é conhecido como o mais perigoso produto já lançado deliberadamente no meio ambiente pelos seres humanos. Sua ação sobre organismos não-alvo, pode levar a alteração no sistema sexual de diversos gastrópodes marinhos, promovendo o surgimento de características masculinas em fêmeas (imposex). Essa alteração se converteu em um excelente biomarcador específico da contaminação marinha por TBT. Nesse sentido, diversos estudos têm mostrado a incidência de imposex em diversos regiões do mundo. Na América do Sul, a Rede Nacional de Estudos em Anti- incrustantes (RNEA) levantou dados referentes a presença de imposex, demonstrando que seus efeitos estão presentes, inclusive dentro de AMPs. Considerando o exposto, e o fato do litoral do Estado de São Paulo possuir diversas áreas de proteção ambiental marinhas (APAs), a ocorrência de imposex em Stramonita brasiliensis, foi avaliada no de duas APAs do litoral paulista. Para isso, foram selecionados 12 pontos, distribuídos no interior das Áreas de Proteção Ambiental Marinhas Central e do Litoral Norte Paulista, considerando as fontes potenciais de TBT para o ambiente, tais como marinas e atracadouros. Em cada ponto, foram coletados 30 indivíduos adultos os quais foram anestesiados e posteriormente avaliados com relação a presença de imposex com base nos índices: % imposex, FPLI, RPLI e VDSI. Todos os locais amostrados, com exceção da praia de Boiçucanga, no município de São Sebastiao, indicaram a ocorrência de imposex que variou de 11 a 100 %. De forma geral, os níveis de imposex foram mais elevados em pontos amostrados nas proximidades de marinas e portos pesqueiros e decresceram com o distanciamento dessas fontes potenciais. Essa observação denota que apesar do banimento global das tintas à base de TBT, esse composto continua a representar um grave problema ambiental para o litoral sul-
americano. Além disso, medidas devem ser tomadas para efetivamente proteger unidades de conservação dos impactos produzidos por substancias químicas perigosas.
57 Butyltin contamination in sediments from Pará state, Amazon Brazil
D.R. Ribeiro, L. Amado, Universidade Federal do Pará UFPA; I.B. de Castro, Universidade Federal de São Paulo / Instituto do Mar; G. Fillmann, Universidade Federal do Rio Grande FURG / Instituto de Oceanografia
TBT-based antifouling paints were banned by the International Maritime Organization in 2008 due at very low concentrations cause gastropod imposex, oyster malformation, and mussel larvae mortality. Given the relatively high logKow of TBT suggests a strong affinity this compound with sediments. The distribution of BT compounds in the sediments samples obtained at river outfalls, fishing ports, shipyards, and industrial zone docks of Pará Harbors (north Brazil) was investigated. Nineteen sediment samples were collected from various locations in 2016 and analyzed for monobutyltin (MBT), dibutyltin (DBT), and tributyltin (TBT) by gas chromatography coupled mass spectrometry (GC-MS). This study is the first report on BT contamination in harbors from Amazon. The Pearson Correlation showed no significant correlation of grain size with TBT, DBT and MBT accumulation. Silt and clay were prevalent in the sediment from Pará state. Higher levels of fine particles (silt) occurred in the saline areas, while clay occurred in sediment samples collected in freshwater. Very fine (clay) to silt were found only in the marine area. The percentage of sediment samples containing coarse fraction were low (< 10%). TBT was present in eight of the nineteen sediment samples collected from the Pará. In this areas TBT concentrations ranged from < LOQ (below limit of quantification) to 27.13 ng Sn/g (harbor). DBT was detected in seven of the stations and varied from 1.89 (P13, beach) to 12.25 ng Sn/g (P4, harbor). MBT was detected only in four sampled sites and ranged from < LOQ to 14.2 ng Sn/g (harbor). The high level of TBT was found in mangrove harbor, while DBT and MBT was found in freshwater harbor. Maximum BTs levels were found in the station freshwater harbor. The results showed that the ratio of TBT to BTs in sediment was generally low, suggests historical inputs organotins, because TBT degradation in aquatic systems may occur way debutylation of TBT to DBT and MBT.
58 Toxicity of common contaminants on aquatic invertebrates in a port area
G.d. Eufrasio Pauly, São Paulo State University - UNESP Campus Experimental do Litoral Paulista; A.F. Cruz, Universidade Estadual Paulista - UNESP/CLP; G. Daniel, UNESP Campus do Litoral Paulista; B.G. Campos, UNESP Campus do Litoral Paulista / Environmental Chemistry; G.L. Altafim, UNESP Campus do Litoral Paulista; D.M. Abessa, Universidade Estadual Paulista - UNESP/CLP / Marine Biology and Coastal Management
Port activities are among the main sources of aquatic contamination, including the active substances of antifouling paints. These booster biocides can be detected in regions close to the marinas, ports and areas where there is intense circulation of vessels. In addition, urban areas and industrial activities are often associated with port activities resulting in the release of several contaminants in the environment, including the ammonia, which has been found in the composition of these discharges. The objective of this study is to evaluate the toxicity of two contaminants, the biocide Irgarol and ammonia, on aquatic invertebrates, namely: embryonic development of the sea urchin Echinometra lucunter, reproduction of the copepod Nitocra sp., and mortality of the nauplii of Artemia salina. E. lucunter embryos were exposed to the concentrations 0.001; 0.01; 0.1; 1.0; and 10.0 mg/L of ammonia and 0.1; 1.0; 10.0; 100.0; and 1000.0 µg/L of Irgarol. Ovigerous females of Nitocra sp. and nauplii of A.
salina were exposed to the concentrations 0.01; 0.1; 1.0; 10.0; and 100.0 mg/L of
ammonia and 1.0; 10.0; 100.0; 1000.0; and 10000.0 µg/L of Irgarol. The assays with
E. lucunter showed LC50-42h of 0.03 mg/L, LOEC of 0.01 mg/L and NOEC of 0.001
mg/L of total ammonia; and no toxicity was observed to Irgarol at the concentrations tested. For the test with Nitocra sp., the observed EC50-7d was 27.89 mg/L, LOEC of
100.0 mg/L, and NOEC of 10.0 mg/L of total ammonia. As for the A. salina nauplii, the value found for the LC50-48h was 368.47 mg/L, LOEC of 1000.0 mg/L, NOEC of
100.0 mg/L of total ammonia; the data relative to the exposure of the nauplii and
Nitocra sp. to Irgarol are being analyzed and will be presented. The nauplii of A. salina
were more resistant to ammonia when compared to the copepods, which had intermediate sensitivity, and sea urchin embryos, which presented higher sensitivity. These results demonstrate that different species may respond differently to different contaminants, requiring a battery of assays with various organisms to assess the ecological risks of chemicals and effluents.
59 Imposex in Thaisella coronata: a study case in Amazon
D.R. Ribeiro, L. Amado, Universidade Federal do Pará UFPA; I.B. de Castro, Universidade Federal de São Paulo / Instituto do Mar; G. Fillmann, Universidade Federal do Rio Grande FURG / Instituto de Oceanografia
Imposex (female masculinization) is induced especially in gastropods exposed to tributyltin (TBT). The purpose of this study was to evaluate the occurrence of imposex in Thaisella coronata collected in port and beach areas from the coast of Pará state (north Brazil – Amazon region). In addition, butyltin levels (TBT, DBT, MBT) in tissue and sediment samples from the same studied areas were also measured. BT levels were determined after derivatization by Grignard reaction using gas chromatography coupled mass spectrometry (GC-MS). Imposex status was determined by RPLI, VDSI and % imposex. The PERMANOVA and T test do not showed differences between port and beach areas considering the butyltin levels in sediments and biota. TBT in sediment was found in two of the three port areas (1.52 ng.g-1 and 2.79 ng.g-1). DBT concentrations ranged from -1 and was found in both
regions, port and beach. This finding indicate the TBT degradation in sediment samples probable induced by solar radiation or microbial activities. In the biota samples TBT concentrations ranged from -¹, DBT ranged from < LOQ to 94.76 ng.g- 1 and MBT ranged from -1. The imposex incidence was observed only in a port area
(100%, RPLI = 1.77). Linear Regression analysis did not show relation of TBT accumulation with imposex indexes. On the other hand, DBTaccumulation strongly relationship with FPLStand and RPLIStand (R² = 0.919, p = 0.0025; R² = 0.868, p =
0.0068, respectively) . This data confirm that imposex is a valid tool to monitor bioavailable BTs, by fact of DBT is a degradation product of TBT. This study reported for the first time the imposex occurrence in Thaisella coronata and suggested this organism as a suitable tool to monitor TBT contamination. Despite statistical analysis do not showed differences between port and beach, the results showed that TBT was generally the most abundant in port sediment. Among six sampled sites, five presented imposex incidence, attributed to ship/boat traffic (probably using TBT-based antifouling in paints) in association to local hydrodynamics conditions.
60 Occurance of Imposex in Strombus pugilis and Melongena melongena gastropods and their relation to contamination sites in the Colombian Caribbean
R. Fernandez-Maestre, University of Cartagena; L.I. Sierra-Marquez, University of Cartagena / Facultad de Ciencias Farmacéuticas; B. Johnson-Restrepo, University of Cartagena / Environmental Chemistry Research Group
Organotin compounds (OTC) are widely used in antifouling paints for boats and surfaces exposed to seawater such as buoys, aquaculture nets, marine ducts and offshore constructions. These paints are used to avoid an undesirable growing of marine organisms (fouling). OTC are also endocrine disruptors that leach from those paints and affect aquatic life causing imposex in gastropods, the growth of a pseudopenis that eventually blocks the female reproductive apparatus, that ends in infertility. Colombian Caribbean have identified several sites contaminated with OTC. The aim of study was to Identifies gastropods (n=173) and relation to imposex from Cartagena city and its surroundings (Cartagena Bay, Baru Island, and Tesca marsh) and the Gulf of Morrosquillo (Caimanera marsh, Covenas beach, and Tolu port). We did not find imposex in Melongena melongena (Linnaeus, 1758) from Tesca marsh (n=26) and Strombus pugilis (Linnaeus, 1758) from Baru Island (n=20) and Covenas beach (n=26) probably due to the small boat traffic in those areas. S. pugilis specimens from Cartagena Bay (n=39) indicated imposex; females showed polifurcated penises up to 15 mm with a standard relative penis size index (RPSI) of 24 and 81% of affected females (%I). M. melongena from Caimanera marsh (n=40) and Tolu port (n=22) showed less imposex than those from Cartagena Bay, RPSI 8.4 and 3.2, and 39%I and 7%I, respectively; However, M. melongena does not seem a good indicator of imposex because a higher RPSI was expected from this port. We found correlation of imposex in gastropods with boat traffic and expected contamination in the Colombian Caribbean probably due to organotin compounds leaching from hull paints. Imposex is an important index to monitor the contamination with endocrine disruptors such as organotin compounds used as antifouling agents in paints for boat hulls.
61 Effects of tributyltin on the eyes, swimming, feeding and growth of newborn guppies Poecilia vivipara
D.V. Paulo, C.F. Mariz Jr, R.M. Barata, UFPE Universidade Federal de Pernambuco / Department of Zoology; P. Carvalho, UFPE - Universidade Federal de Pernambuco / Zoología
Tributyltin (TBT) is an antifouling biocide highly toxic to non-target aquatic organisms. Newborns of Poecilia vivipara obtained from breeding stock kept in the laboratory were exposed for 96 h to tributyltin (TBT) in treatments 0 (control), 0 (solvent control - dimethylsulfoxide); 0.01; 1.0; 5.0; 7.5 and 10 µg L-1 of TBT. Ten
fish with six days of life, 7 mg of weight and 8,9 mm of total length were used at the beginning of exposures. After exposure, we evaluated the swimming speed of the individual fish by tracking videos, swimming resistance by standardized counter- current swimming, quantitative measures of the ability to capture Artemia nauplii, growth in weight and length, and histology of the eyes and retina. Macroscopic
analysis of the eyes showed a darkening of the iris region after exposure to 5.0; 7.5 and 10.0 µg TBT L-1. Histopathological analyzes revealed a pigment compaction in
the pigment epithelium layer of the retina and in the iris, with degeneration of retinal epithelial cells. In addition, changes in the photoreceptor layer were also observed in fish exposed to 5.0; 7.5 and 10.0 µg TBT L-1. Significant reductions relative to controls
of 85% in the mean swimming speed, 60% in the swiming resistance index, and of 33.6% in the daily catch of Artemia nauplii throughout the exposure were detected at 7.5 µg TBT L-1. In agreement with the reduction in prey catch, a reduction in the
growth of fish exposed to 7.5 µg L-1 of 56.3% in biomass and 64.3% in standard length
was detected. The histopathological changes detected in the retina and iris at 5.0; 7.5 and 10.0 µg TBT L-1 may have reduced the fish visual exploration and prey detection
capacity, which together with the detected reduction in swimming speed, might have led to a deficiency in prey capture. This was corroborated by the reduction in prey capture observed, culminating in the growth reduction of P. vivipara newborns. These changes favor the possibility of higher predation risk of the exposed fish, and reduce the chances of these fish recruiting to the adult population.
62 Does Sediment Organic Carbon Influence the Toxicity of the Biocide Chlorotalonil?
M.A. Ferraz, Centro de Estudos do Mar da Universidade Federal do Paraná; T. Santin, V. Calabresi, Instituto do Mar da Universidade Federal de São Paulo; R.B. Choueri, Unifesp Universidade Federal de São Paulo / Departamento de Ciências do Mar; I.B. de Castro, Universidade Federal de São Paulo / Instituto do Mar; F. Gallucci, Federal University of Sao Paulo / Marine Sciences
Chemical factors such as sediment redox potential, salinity, pH and organic carbon content interfere in the stability of apolar organic compounds, making them more or less available to the biota. Contaminants with low solubility in particular, as the case of the antifouling biocide Chlorotalonil, exhibit a sortive affinity to the organic carbon present in sediments. As such, is expected that biocide levels available in the interstitial water are inversely related to organic carbon amounts in sediment. In this study, a microcosm experiment using a natural meiobenthic community was performed to investigate the influence of sediment organic carbon content in the toxicity of Chlorotalonil. Meiofauna communities were exposed for 7 days to three environmental relevant concentration of Chlorotalonil (0.01 µg.g-1, 0.1 µg.g-1 and 0.2
µg.g-1) in sediments with different organic carbon content (0.1% and 0.4%) and were
compared to non-contaminated treatments. In sediments presenting low levels of organic carbon, the densities of total meiofauna, nematodes and ostracodes were significantly lower in treatments contaminated with moderate and high concentrations comparing to the controls. Multivariate structure of nematode assemblages was also significantly different in contaminated treatments. On the other hand, sediments with lower organic carbon amounts showed no differences in meiofauna univariate and multivariate descriptors between contaminated and control treatments. The differential response of the community to contamination by Chlorotalonil in the two different sediments clearly demonstrates the influence of organic carbon on toxicity. It further suggests a higher bioavailability of the contaminant for meiofauna in sediments slightly enriched. Since Chlorotalonil has a tendency to partition onto organic matter, it is possible that contamination of meiofauna was mainly through the ingestion of contaminated particles, rather than through the ingestion and/or absorption of interstitial water, as it has been previously suggested.
63 Efeitos do clorotalonil sobre o sistema imune de mexilhões Perna perna
A.d. Guerreiro, R.C. Rola, Universidade Federal do Rio Grande FURG / Instituto de Ciências Biológicas ICB; M.T. Rovani, Universidade Federal de Pelotas / Faculdade de Veterinária; S.R. Costa, FURG Universidade Federal do Rio Grande / Instituto de Ciências Biológicas ICB; J. Sandrini, Universidade Federal do Rio Grande - FURG / Instituto de Ciências Biológicas - ICB
As tintas anti-incrutantes têm sido amplamente utilizadas para fins de proteção de embarcações contra incrustações marinhas. Muitos biocidas que compõem essas tintas surgiram como alternativas ao uso do TBT, um composto que exerce exacerbada toxicidade para diversos organismos não alvo, tais como ostras Crassostrea gigas e mexilhões Perna perna. O biocida clorotalonil, por exemplo, começou a ser utilizado na composição de tintas anti-incrustantes depois do banimento do TBT e seus efeitos sobre a fauna marinha ainda são parcialmente desconhecidos. Nesse sentido, o presente trabalho teve como objetivo avaliar os efeitos do clorotalonil sobre parâmetros do sistema imunológico de mexilhões P. perna. Para isto, animais foram expostos a 0 (controle, com 0,016 % do solvente dimetil sulfóxido), 0,1 µg/L e 10 µg/L de clorotalonil por 96 h. Após 24 h e 96 h, a hemolinfa foi retirada e os seguintes parâmetros imunológicos foram analisados nos hemócitos: número total de hemócitos, geração de espécies reativas de oxigênio, viabilidade celular (MTT e vermelho neutro), atividade fagocítica, adesão celular e dano de DNA. Tanto após 24 h quanto
96 h, pode-se observar que a viabilidade celular foi diminuída, enquanto que a adesão celular foi aumentada. Além disto, após 96 h de exposição ao contaminante, um aumento na atividade fagocítica também pode ser observado em ambas concentrações testadas. O número de células presentes na hemolinfa, o nível de espécies reativas de oxigênio e o dano de DNA, entretanto, não foram afetados. Sendo assim, os resultados encontrados no presente trabalho sugerem que o clorotalonil é um biocida que afeta o sistema imune de organismos não alvo, o que pode, inclusive, afetar a capacidade destes animais em responder a outros estressores ambientais de maneira eficiente.
64 Exposure to chlorothalonil causes change in biomarkers of contamination in the estuarine polychaete Laeonereis acuta
J. da Silva Barreto, F.d. Tarouco, Universidade Federal do Rio Grande FURG; F.A. de Godoi, Universidade de Sao Paulo; M. Geihs, Universidade Federal do Rio Grande FURG; J. Sandrini, Universidade Federal do Rio Grande - FURG / Instituto de Ciências Biológicas - ICB; C. da Rosa, Universidade Federal do Rio Grande FURG / Instituto de Ciências Biológicas
Chlorothalonil is a broad spectrum fungicide applied in agricultural activities with the purpose to fight against foliar and crop diseases, in addition to currently being widely used as an active biocide in antifouling paints. Thus, the aim of this study was to evaluate the effects of chlorothalonil exposure on biochemical biomarkers of oxidative metabolism as well as cholinesterases in the estuarine polychaete Laeonereis acuta. For this, the activity of the enzymes catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), glutamate cysteine ligase (GCL), acetylcholinesterase (AChE) and propionylcholinesterase (PChE) were evaluated as well as the levels of reduced glutathione (GSH), antioxidant capacity and levels of lipid peroxidation. After acclimatization the animals were exposed for 24 and 96 h at the following nominal concentrations of chlorothalonil: 0.1, 10.0 and 100.0 µg/L, originated from stock solutions prepared on DMSO. No difference was observed in the levels of reactive oxygen species in animals exposed to chlorothalonil. However, there was a reduction of the antioxidant capacity against peroxyl radicals of the animals exposed to the highest concentration after 96 h concomitantly with an increase in the levels of lipid peroxidation. Considering the components of the antioxidant defense system, no differences were observed in the activity of CAT and SOD enzymes at both experimental times. However, the activity of the GCL enzyme was increased after 24 h of exposure to the highest concentration, as well as an increase in GST activity after 96 h in this same treatment. GSH levels were increased in the animals exposed to the lowest concentration, and reduced in the highest concentration after 96 h. Regarding the intermediate metabolism, a reduction was observed in lactate levels at 10µg/L after 24 h, while the higher concentration induced an increase in lactate levels after 96 h. There were no differences in glycogen levels at any of the exposure times. AChE activity was inhibited in the animals exposed to the highest concentration after 96 h. These results demonstrate that chlorothalonil was toxic to L.
acuta, altering its metabolism and causing a situation of oxidative stress in high
concentrations after 96 h of exposure. In addition, it has been shown its capacity to alter the activity of enzymes of the cholinergic system, which are widely used in biomarkers studies.