Identification of microcystins in waters used for daily life by people who live on Tai Lake during a serious cyanobacteria dominated bloom with risk analysis to human health

Tai Lake is the third largest freshwater lake in China with annual cyanobacteria blooms. Microcystins produced by these blooms have serious health risks for populations surrounding the lake, especially for people living on Tai Lake, because they usually drink raw lake water after a simple alum treatment. This study presents data on the detection and identification of microcystins in waters used for daily life by people living on Tai Lake, during the cyanobacterial blooming in July 2007. The health risks from drinking these microcystin-polluted waters were also calculated. The main microcystins detected by high-performance liquid chromatography-electrospray ionization mass spectrometry in the water samples collected from two parts of Tai Lake (Wuli Lake and Meiliang Bay) were MC-LR (4.33-12.27 microg/L), MC-RR (8.36-16.91 microg/L) and MC-YR (1.41-5.57 microg/L). Risk assessment showed that the drinking water simply treated by alum was not safe. The lowest calculated hazards ratios in all water samples was 6.4, which indicated that the risk of microcystins exposure from drinking water was over six times higher than the tolerable daily intake (TDI) recommended by The World Health Organization (WHO). Further studies should be conducted to elucidate the relationships between the epidemiology of people living on Tai Lake and microcystins exposure from drinking water.     

Variations in the Microcystin Content of Different Fish Species Collected from a Eutrophic Lake

Microcystins produced from cyanobacteria can accumulate in fish tissues. Liquid chromatography coupled with tandem quadrupole mass spectrometry (LC-MS/MS) is an attractive alternative to immunoassays for the determination of low concentrations of microcystins in tissues. Fish taken from Grand Lake St. Marys, a eutrophic lake in Ohio, USA, were analyzed for microcystin-LR in their fillets using LC-MS/MS. Of 129 fish tested for microcystins, only black crappie (Pomoxis nigromaculatus) and common carp (Cyprinus carpio) tested positive for microcystin-LR. Less than 10% of Pomoxis and 7% of Cyprinus samples contained measurable levels of microcystin-LR. Statistical analysis yielded a p-value of 0.07 between Pomoxis and the pooled results of the other four fish species. However, this comparison was complicated by the large difference in sample size between species. Further sampling in Grand Lake St. Marys for microcystin-LR would help determine if microcystin-LR exposure occurs through foodweb transfer.     

Toxicological profile of pollutants in surface water from an area in Taihu Lake, Yangtze Delta

The environment in urbanized areas is often contaminated with a complex mixture of toxic compounds originating from industries, agriculture and private households. Most of the contaminants end up in surface waters, such as lakes, rivers and finally the sea. Toxic contaminants may disturb the biological condition of aquatic ecosystems and be harmful for humans, if they are transported to human food or drinking water. A variety of biological tests have been introduced for monitoring the toxicological profile of aquatic ecosystems. In the present investigation, genotoxic, hormone disrupting and Ah-receptor activity was analyzed in water collected in January 1999 at Meiliang Bay, Taihu Lake, China, near to the city of Wuxi in the Shanghai area. Significant mutagenic activity could be detected in water extracts with the Salmonella/microsome assay and the arabinose resistance test. Frame shift mutations were the predominant mode of action. Ah-receptor active compounds were detected by using a luciferase reporter gene assay (CALUX-assay). The estimated toxic equivalent factor-values ranged between 134 and 232 pg TCCD-equivalents per liter lake water. The estrogen-like potential of Taihu water was estimated with two luciferase reporter gene assays using transgenic human cell lines expressing estrogen receptor alpha. Estradiol equivalents obtained with water extracts were in the range of 2.2-12.1 ng/l. We also analyzed the concentrations of 17-beta-estradiol and ethinylestradiol in the extracts using a high-pressure liquid chromatography-method. The values obtained correlated with the biologically determined estradiol equivalents, indicating that the estrogenic activity is mainly due to natural and synthetic hormones rather than xenoestrogens.     

Microalga Euglena as a bioindicator for testing genotoxic potentials of organic pollutants in Taihu Lake,China

The microalga Euglena was selected as a bioindicator for determining genotoxicity potencies of organic pollutants in Meiliang Bay of Taihu Lake, Jiangsu, China among seasons in 2008. Several methods, including the comet assay to determine breaks in DNA and quantification of antioxidant enzymes were applied to characterize genotoxic effects of organic extracts of water from Taihu Lake on the flagellated, microalga Euglena gracilis. Contents of photosynthetic pigments, including Chl a, Chl b and carotenoid pigments were inversely proportion to concentrations of organic extracts to which E. gracilis was exposed. Organic extracts of Taihu Lake water also affected activities of superoxide dismutase (SOD) and peroxidase (POD) of E. gracilis. There were no statistically significant differences in SOD activities among seasons except in June but significant differences in POD activities were observed among all seasons. The metrics of DNA fragmentation in the alkaline unwinding assay (Comet assay), olive tail moment (OTM) and tail moment (TM), used as measurement endpoints during the genotoxicity assay were both greater when E. gracilis was exposed to organic of water collected from Taihu Lake among four seasons. It is indicated that the comet assay was useful for determining effects of constituents of organic extracts of water on E. gracilis and this assay was effective as an early warning to organic pollutants.     

DNA damage caused by extracts of chlorinated drinking water in human derived liver cells (HepG2)

Dong (D) lake and the Yangtze (Y) river are the main water supplies of the city of Wuhan, PR China. In the present study, the genotoxic effect of chlorinated drinking water (CDW) processed from raw water of D lake and Y river was evaluated in human HepG2 cells using the Comet assay and the micronucleus test. For that, HepG2 cells were exposed to XAD extracts of CDW corresponding to 0.167, 1.67, 16.7 and 167 ml CDW/ml cell culture. All CDW extracts caused a significant and dose-dependent increase of DNA migration in HepG2 cells. The level of DNA damage varied depending on the sampling time (season) and sampling site. The lowest concentration which caused a significant increase of DNA migration was 1.67 ml CDW/ml culture for water samples collected in August. Water samples collected in March showed their lowest observable effect levels in 167 ml and 16.7 ml CDW/ml culture for Y river and D lake, respectively. Additionally, significant increases of micronuclei (MN) frequencies were found in HepG2 cells after CDW treatment. However, in the MN assay the CDW samples collected in March exhibited higher genotoxicity than the August samples. In conclusion, HepG2 cells provide a useful tool for the detection of genotoxic effects of environmental mixtures.     

An ultra-performance liquid chromatography–tandem mass spectrometry method for determination of microcystins occurrence in surface water in Zhejiang Province, China

A novel method had been developed for determination of microcystin in surface water by ultra-performance liquid chromatography combined with tandem mass spectrometry (UPLC–MS/MS). Water samples after being concentrated and cleaned with solid phase extraction (SPE) with Oasis HLB cartridge were separated on AcQuity UPLC™ BEH C18 column (1.7 μm, 1.0×50 mm) with a gradient elution and a mobile phase consisting of 0.1% formic acid in water and methanol, and then detected with an electrospray ionization tandem mass spectrometry in positive ion mode with multiple reaction monitoring. Compared with traditional liquid chromatography, it took much less time for UPLC to analyze microcystin. Additionally, the UPLC–MS/MS method produced satisfactory reliability, sensitivity, and accuracy. Recoveries of the four microcystins were 91.7–111% with relative standard deviation being 7.9–12%. The calibration curves for microcystins were linear with correlation coefficient being larger than 0.99. Limit of quantification of the four microcystins (microcystin-LR, RR, LW, and LF) after 1000-fold SPE pre-concentration, were 2.5, 6.0, 2.5, and 1.3 ng/L. The presented method had been successfully used to detect microcystin in surface water from the Qiantang River, the West Lake, and three drinking water reservoirs, which represented water bodies of river, lake, and drinking water reservoir, respectively. It was shown that microcystin pollution in drinking water reservoirs was the heaviest, and microcystin-LR and RR were the predominant kinds, with concentrations ranging 0.045–2.73 and 0.021–1.36 μg/L, respectively. The concentrations of microcystins in river and lake were all under detection limit.     

Genotoxicity of crude extracts of cyanobacteria from Taihu Lake on carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>)

Genotoxicity of crude cyanobacteria extracts (CBE) from blooms in Taihu Lake, China toward common carp (Cyprinus carpio) was measured. The primary extracellular product was determined by HPLC to be Microcystin-LR (MC-LR, L for leucine and R for arginine) with an average concentration of 2.4 × 10² μg MC g⁻¹ dry weight of cyanobacteria. Acute toxicity to carp, expressed as the 72-h LC_50, was 53 mg, dw cyanobacteria L⁻¹. Genotoxicity, as determined by the micronucleus (MN) and comet assays were both dose- and time-depended. Deformities of cellular organelles in liver and gill were observed by use of transmission electron microscopy (TEM). The results showed that MC-LR from cyanobacteria from Taihu Lake could induce genotoxic response and tissue-level morphological changes in common carp.     

Genotoxicity study with special reference to DNA damage by comet assay in fission yeast, Schizosaccharomyces pombe exposed to drinking water.

The objective of this study was to investigate genotoxicity, especially DNA damage, in drinking water samples collected from tap by using fission yeast Schizosaccharomyces pombe as a model organism. Generally raw water potabolization is done by treatment with polymeric coagulant, alum, chlorine, etc. In the comet test, highly significant (P<0.001) effects of DNA damage were detected in treated water (tap water) when compared to negative control (raw water) as well as laboratory control (distilled water) samples for both 1 h and 2 h exposure. In the water treatment plant, raw water treatment is done by the process of prechlorination, alum and polymeric coagulant (CatflocT) dosing, postchlorination, filtration and final discharge for consumption. In conclusion it can be stated from the results that chlorinated disinfectant, alum and polymeric coagulant (CatflocT) mixture used in drinking water has a potent cumulative genotoxic effect in the eukaryotic cells and may pose potential genotoxic risk for human health following long-term consumption.     

Spatiotemporal Dynamics of Microcystin Variants and Relationships with Environmental Parameters in Lake Taihu,China

Excessive anthropogenically-caused nutrient loading from both external and internal sources has promoted the growth of cyanobacteria in Lake Taihu from 2005 to 2014, suggesting increased production and release of cyanotoxins. In order to explain the spatial distribution and temporal variation of microcystins (MCs), the intracellular concentrations of MCs (MC-LR, -RR and -YR, L, R and Y are abbreviations of leucine, arginine and tyrosine) were monitored monthly from July 2013 to June 2014. Three MC variants are present simultaneously in Lake Taihu; the MC-LR and -RR variants were dominant (accounting for 40% and 39% of the total), followed by MC-YR (21%). However, MC-YR accounted for a higher proportion in colder months, especially in March. The highest concentrations of intracellular MCs were found in July and October when cyanobacteria cell density also reached the maximum. The average concentrations of MC-LR, -RR and -YR in July were 4.69, 4.23 and 2.01 μg/L, respectively. In terms of the entire lake, toxin concentrations in northern parts were significantly higher than the eastern part in summer, when MC concentrations were several times higher than the guideline value by WHO throughout much of Lake Taihu. Results from correlation and redundancy analysis (RDA) showed that total MCs, including all variants, were strongly and positively correlated with cyanobacteria cell density, water temperature, total phosphorus (TP) and pH, whereas each variant had different correlation coefficients with each of the considered environmental variables. MC-RR showed a stronger relationship with temperature, in contrast to MC-YR and -LR. Dissolved inorganic carbon (DIC) showed a negative relationship with each variant, suggesting that rising DIC concentrations may inhibit cyanobacterial growth and thereby reduce MC production in the future.     

Assessment of estrogenic contamination and biological effects in Lake Taihu

Lake Taihu is the third largest freshwater lake in China and is contaminated with xenoestrogens associated with high population density, intensive livestock and aquatic breeding activities. A field study in Lake Taihu was conducted using the goldfish (Carassius auratus) as an indicator organism. Several biological markers were selected to assess the extent of estrogenic contamination. Changes in serum vitellogenin (VTG), and gill 7-Ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST) and reduced glutathione (GSH) were measured in caged juvenile goldfish for 28 days in seven locations in northern Lake Taihu. Bioassay showed VTG increased 0.64–2.42 folds over time in goldfish collected from five stations and GSH decreased in samples from all seven stations after 7 days of exposure. EROD levels increased continually in fish collected at all the seven stations and the highest concentrations occurred at day 21. GST activity increased significantly at 7 days. The concentration of the target estrogens estrone (E₁), 17β-estradiol (E₂), ethinylestradiol (EE₂), octylphenol (OP), diethylstilbestrol (DES), nonylphenol (NP) and bisphenol A (BPA) were determined in lake water at the sampling stations. Each individual estrogen concentration measured was multiplied by its relative potency to gain the estrdiol equivalent (EEQ). There was an obvious correlation between the concentration of VTG and the total EEQ for all seven locations (P < 0.001). The biomarker VTG, EROD, GST and GSH assays and chemical analysis might be used to illustrate the potential risk in Lake Taihu.     

Use of four short-term tests to evaluate the mutagenicity of municipal water

Some ways in which four short-term tests may be used to evaluate the mutagenicity of drinking water were explored by testing raw and treated water from Lake Bloomington, which serves the town of Bloomington, Illinois (population, 44,000). The water was collected from February 1976 to October 1977 and was concentrated by evaporation or by use of XAD-2 resin. The water was tested for the ability to induce reverse mutation in a prokaryote, Salmonella typhimurium; forward mutation in a mold, Neurospora crassa; mitotic gene conversion in a yeast, Saccharomyces cerevisiae; and reverse mutation in maize, Zea mays. Because of the large number of water samples (54) and the limited amounts of the samples, it was not possible to test all samples in all four tests by all the protocols. Thus, the sensitivities of the four tests to potential mutagens in the water samples could not be rigorously compared. However, the results do show that lake and tap water samples collected during 1976 were toxic but not mutagenic in N. crassa and neither toxic nor genotoxic in S. cerevisiae; lake water collected during 1977 was mutagenic in one line of Z. mays and slightly mutagenic in S. typhimurium strain TA1536 in the presence of rat liver S9. The results suggest that tests that detect a variety of genetic end points should be used when testing complex mixtures such as drinking water. The advantages and disadvantages of the tests and protocols are discussed in terms of their applicability to the study of the mutagenicity of drinking water.     

Microcystin concentrations and genetic diversity of Microcystis in the lower Great Lakes

The resurgence of Microcystis blooms in the lower Great Lakes region is of great concern to public and ecosystem health due to the potential for these colonial cyanobacteria to produce hepatotoxic microcystins. A survey of Microcystis cell densities and microcystin concentrations during August 2004 showed particularly high concentrations of both cells and toxin in the nearshore regions of Saginaw Bay (Lake Huron) and western Lake Erie, often exceeding the World Health Organization's recommended drinking water limit of 1 microg L(-1). The dominant congener of microcystin in both basins was microcystin-LR (MC-LR), whereas the second most abundant congeners, accounting for up to 20-25% of the total microcystin concentrations, were MC-LA in Saginaw Bay and MC-RR in western Lake Erie. Multiplex PCR assays of Microcystis colonies isolated from these two regions showed that a much greater percentage of the Microcystis colonies from Saginaw Bay carried the mcyB gene necessary for microcystin production, in comparison with those from western Lake Erie. The mcyB genotypes sequenced separated into two distinct phylogenetic clusters, with Microcystis originating from Lake Erie predominantly in one branch and from Saginaw Bay present in both branches. These results indicate that the genetic composition of the bloom could impact the concentrations and congeners of microcystin produced and that the cell count methods currently being used to gauge public health threats posed by Microcystis blooms may not sufficiently assess actual bloom toxicity.     

First Report on Microcystin-LR Occurrence in Water Reservoirs of Eastern Cuba,and Environmental Trigger Factors

The factors related to cyanotoxin occurrence and its social impact, with comprehension and risk perception being the most important issues, are not yet completely understood in the Cuban context. The objectives of this research were to determine the risk extension and microcystin-LR levels, and to identify the environmental factors that trigger the toxic cyanobacteria growth and microcystin-LR occurrence in 24 water reservoirs in eastern Cuba. Samplings were performed in the early morning hours, with in situ determination and physicochemical analysis carried out in the laboratory. Microcystin-LR were determined in water and within the cells (intracellular toxins) using UPLC–MS analysis after solid phase extraction. The reservoirs studied were found to be affected by eutrophication, with high levels of TN:TP ratio and phytoplankton cell concentrations, high water temperatures and low transparency, which cause collateral effect such as cyanobacterial bloom and microcystin-LR occurrence. In Hatillo, Chalóns, Parada, Mícara, Baraguá, Cautillo, La Yaya, Guisa and Jaibo reservoirs, concentrations of MC-LR higher than the WHO limits for drinking water (1 µg·L⁻¹), were detected.     

Simplified protocol for evaluating the genotoxic risk of hospital wastewater

To evaluate the genotoxicity of hospital wastewater, we drew up a simplified protocol based on two well-known tests: the SOS chromotest and the Ames fluctuation test. Three concentrations of wastewater samples were directly tested without extraction or concentration. By fixing three significance levels in genotoxicity response for each test, we could classify the samples in five categories ranging from nongenotoxic to highly genotoxic. This simplified classification thus constitutes an invaluable help in making the nonscientist public aware of the genotoxic risk of hospital wastewater and can be applied in a screening approach to chemicals in the environment.     

Health risk of semi-volatile organic pollutants in Wujin river inflow into Taihu Lake

Semi-volatile organic compounds (SVOCs), including polycyclic aromatic hydrocarbons (PAHs), phthalates and pesticides in water of Wujin river inflow into Taihu Lake, were detected for assessment of risk to human health. SVOCs were tested with Method 525.2 established by the US Environmental Protection Agency (EPA) and health risk assessment (HRA) was conducted by the hazard quotient (HQ) approach from US HRA for screening stage. The results for the liquid–solid extraction of water sample measured by gas chromatography/mass spectrometry (GC/MS) showed that three of the tested twenty-three semi-volatile organic contaminants posed potential health risk and the concentrations of 2,4-dinitrotoluene, di-n-butyl phthalate, chrysene and benzo(a)anthracene was 0.736, 15.201, 0.307, and 0.334 μg/l, respectively. SVOCs in the Wujin river water might induce risk to environmental health of Taihu Lake as a source of drinking water.     

Assessing the toxicity of ingested Taihu Lake water on mice via hepatic histopathology and matrix metalloproteinase expression

Water from Taihu Lake (China) is used as a drinking source. The frequency of contamination in Taihu Lake has increased over the past decade and the bloom-forming cyanobacteria are the dominant species during eutrophication. Cyanobacteria can produce various harmful secondary substances including microcystins capable of endangering human health and ecological safety. This study investigated toxicity of ingested Taihu Lake water on mice via hepatic histopathology and matrix metalloproteinase (MMP) expression. Water was sampled from four Taihu Lake locations, Meiliang Bay 2 group (M2), Meiliang Bay 1 group (M1), Lake Center (H) and Xukou Bay (X), along a gradient of decreasing degree of eutrophication. The experimental design consists of five groups of male mice (Mus musculus, ICR): one control and four groups ingesting water from the four sampling sites for 90 days. Compared to control, M1 and M2 mice showed hepatic histopathological changes including swollen, vacuolar degeneration or inflammatory. Immunohistochemical staining demonstrated a higher expression of MMP-2 proteins in M2 group and a lower expression of MMP-9 in M1. Enzyme-linked immunosorbent assay indicated that MMP-9 concentration was significantly reduced from 0.55 to 0.28 ng/g liver weight in M2 (p < 0.05). Real time PCR revealed a down-regulation of MMP-9 mRNA by 2.2 fold in M1 and an up-regulation of MMP-2 mRNA by 1.73 fold in H. Using this mouse model as a gauge of water toxicity, our results revealed that potential health risks induced by Taihu Lake water might arise from the use of this source water by local resident.     

Preliminary evaluation of gene expression profiles in liver of mice exposed to Taihu Lake drinking water for 90 days

Differential gene expression profiling was performed via DNA microarray in the liver tissue of Mus musculus mice after exposure to drinking water of Taihu Lake for 90 days. A total of 75 differentially expressed candidate genes (DEGs) were identified (1.5-fold, p ≤ 0.05), among which the expression of 29 genes was up-regulated and that of 46 genes was down-regulated. Most DEGs were involved in biological process based on gene ontology mapping analysis. The drinking water of Taihu Lake significantly influenced the expression of genes related to cell proliferation, apoptosis, amino acid metabolism, development and immune responses. Long-term exposure to the Taihu drinking water may result in increased carcinogenic risk.     

Contamination by metals and pharmaceuticals in northern Taihu Lake (China) and its relation to integrated biomarker response in fish

Taihu Lake is the largest shallow freshwater lake in eastern China and is suffering not only from an increasingly serious threat of eutrophication but also potential ecological risk due to the input of emerging contaminants. Active biomonitoring was conducted in Taihu Lake using transplanted goldfish (Carassius auratus) to determine the contamination by pharmaceuticals and metals and to assess the potential ecological risk. A suite of biomarkers including acetylcholinesterase, ethoxyresorufin O-deethylase, glutathione S-transferase, glutathione peroxidase and superoxide dismutase activities in fish after 7, 14, 21 and 28 days of exposure in situ, as well as pharmaceuticals and metals in water, were determined during the field exposure period. The results indicate that pharmaceuticals exist mainly in Zhushan Bay and Meiliang Bay, while metals are present mainly in Gong Bay. An integrated biomarker response (IBR) was calculated and used to evaluate the ecological risk of the polluted area of Taihu Lake. It was found that Zhushan Bay might present higher risk to fish, followed by Meiliang Bay. IBR values were in good agreement with copper and caffeine concentrations.     

Changes in concentrations of microcystins in rainbow trout, freshwater mussels, and cyanobacteria in Lakes Rotoiti and Rotoehu

Microcystin concentrations in cyanobacteria and their accumulation in rainbow trout (Oncorhynchus mykiss) and freshwater mussels (Hyridella menziesi) in Lakes Rotoiti and Rotoehu (New Zealand) were investigated. Hatchery rainbow trout were added to an enclosure in Lake Rotoiti where concentrations of microcystins in the phytoplankton and cyanobacterial cell concentrations could be closely monitored. Rainbow trout that were free to roam in the entire area of each lake were also included in the study. Freshwater mussels were suspended subsurface in cages in the enclosure. Phytoplankton samples, rainbow trout liver and muscle tissue, and the tissues of mussels were analyzed for microcystins using the ADDA-ELISA method, and selected samples were analyzed using LC-MS. A maximum concentration of microcystins in the phytoplankton samples of 760 microg L(-1) was recorded in Te Weta Bay, Lake Rotoiti, in March 2004. ELISA results confirmed microcystin immunoreactivity in rainbow trout liver and muscle tissues and in freshwater mussels. The microcystin congeners LR, YR, RR, AR, FR, LA, and WR were detected by LC-MS in caged freshwater mussels in Lake Rotoiti but were not detected in either muscle or liver tissue of rainbow trout. The daily tolerable intake limit of microcystins for human consumption recommended by the World Health Organisation is 0.04 microg kg(-1) day(-1). Modeling was carried out for the human intake of microcystin compounds from rainbow trout muscle tissue, and the potential health risks were estimated, assuming the ADDA-ELISA was determining compounds of toxicity equivalent to microcystin-LR.     

Microcystins (cyanobacteria hepatotoxins) bioaccumulation in fish and crustaceans from Sepetiba Bay (Brasil, RJ).

Blooms of cyanobacteria in water bodies cause serious environmental problems and the occurrence of toxic strains are also related with the human health. Aquatic animals could bioaccumulate microcystins (cyanobacteria hepatotoxins) and so, beyond water, the ingestion of contaminated food represents a human health risk. Recently, WHO recommended a maximum concentration of microcystins (MCYSTs) in drinking water and established the tolerable daily intake (TDI) for consumption of cyanobacteria products contends MCYSTs (0.04 microg(-1) kg(-1) day(-1)). Sepetiba Bay is located in the municipal districts of Rio de Janeiro, Mangaratiba and Itaguai; being an important place of fishing activity. Due to the industrial development in the area, this bay is submitted to different environmental impacts, increasing the organic and industrial pollution. A strain of the nanoplanktonic cyanobacteria Synechocystis aquatilis f. aquatilis that produce MCYSTs was already isolated. In this study, we verified MCYSTs presence in muscle tissue of fish and crustaceans, which were harvested monthly in Sepetiba Bay during 11 months, in order to evaluate the potential risk of their ingestion. MCYSTs were analyzed by immunoassay techniques using the ELISA Microcystin Plate Kit (ENVIROLOGIX INC) and the concentration were expressed as microcystin-LR equivalent. The analyses of seston samples, water, muscle tissues showed the presence of this cyanotoxin in all samples and it was verified that 19% of the animals' samples were above the limit recommended by WHO for human consumption. The maximum value found was of 103.3 microg kg(-1) (TDI 0.52 microg kg(-1) day(-1)) and the minimum, was 0.25 microg kg(-1) in crabs muscle tissue (TDI of 0.001 microg kg(-1) day(-1)). Such data demonstrate that, although in low concentrations, there is already a contamination of fish and crustaceans from Sepetiba Bay. We highlight that the recommended limit refers to healthy adult.