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BACKGROUND: Coal and coal ash present inorganic elements associated with negative impacts on environment and human health. The objective of this study was to compare the toxicity of coal and coal ash from a power plant, assess their inorganic components, and investigate the biological impacts and potential mechanisms through in vitro and in vivo testing. METHODS: Particle-Induced X-ray Emission method was used to quantify inorganic elements and the toxicity was evaluated in Caenorhabditis elegans and Daphnia magna in acute and chronic procedures. The genotoxic potential was assessed using alkaline and FPG-modified Comet assay in HepG2 cells and mutagenicity was evaluated using Salmonella/microsome assay in TA97a, TA100, and TA102 strains. RESULTS: Inorganic elements such as aluminum (Al) and chromium (Cr) were detected at higher concentrations in coal ash compared to coal. These elements were found to be associated with increased toxicity of coal ash in both Caenorhabditis elegans and Daphnia magna. Coal and coal ash did not induce gene mutations, but showed genotoxic effects in HepG2 cells, which were increased using the FPG enzyme, indicating DNA oxidative damage. CONCLUSIONS: The combined findings from bioassays using C. elegans and D. magna support the higher toxicity of coal ash, which can be attributed to its elevated levels of inorganic elements. The genotoxicity observed in HepG2 cells confirms these results. This study highlights the need for continuous monitoring in areas affected by environmental degradation caused by coal power plants. Additionally, the analysis reveals significantly higher concentrations of various inorganic elements in coal ash compared to coal, providing insight into the specific elemental composition contributing to its increased toxicity.

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Exposure of tobacco workers handling dried tobacco leaves has been linked to an increased risk of toxicity and respiratory illness due to the presence of nicotine and other chemicals. This study aimed to evaluate the DNA damage caused by the exposure of tobacco growers during the dry leaf classification process and the relation to cellular mechanisms. A total of 86 individuals participated in the study, divided into a group exposed to dry tobacco (n = 44) and a control group (n = 42). Genotoxicity was evaluated using the alkaline comet assay and lymphocyte micronucleus (MN) assay (CBMN-Cyt), and measurement of telomere length. The levels of oxidative and nitrosative stress were evaluated through the formation of thiobarbituric acid reactive species, and nitric oxide levels, respectively. The inorganic elements were measured in the samples using particle-induced X-ray emission method. The combination of variables was demonstrated through principal component analysis and the interactions were expanded through systems biology. Comet assay, MN, death cells, thiobarbituric acid reactive species, and nitrosative stress showed a significant increase for all exposed groups in relation to the control. Telomere length showed a significant decrease for exposed women and total exposed group in relation to men and control groups, respectively. Bromine (Br) and rubidium (Rb) in the exposed group presented higher levels than control groups. Correlations between nitrate and apoptosis; Br and MN and necrosis; and Rb and telomeres; besides age and DNA damage and death cells were observed. The systems biology analysis demonstrated that tobacco elements can increase the nuclear translocation of NFKB dimers inducing HDAC2 expression, which, associated with BRCA1 protein, can potentially repress transcription of genes that promote DNA repair. Dry tobacco workers exposed to dry leaves and their different agents showed DNA damage by different mechanisms, including redox imbalance.

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BACKGROUND: The elemental profile of the Marselan wine cycle was studied with a single ion-beam technique, namely particle-induced X-ray emission (PIXE). Samples were collected from all of the major stages involved in the wine cycle, including soil, vine plant, grape, and different stages of wine processing. RESULTS: The results show the influence of field practices on the elemental composition of leaves, branches, grapes, and soil. Mechanisms of element precipitation and elimination from the wine were identified through the changes in the elemental concentrations measured during the post-harvesting processes. The concentrations of some elements, such as Al, Si, K, and Ni, did not vary for grapes collected at different maturation stages. On the other hand, changes in the concentrations of Si, Cl, Ca, Cu, and K were observed right after the maceration and pressing processes in young wine. Finally, the reduction in the Cu concentration during the last stages of the wine cycle is related to its precipitation in the storage tanks. CONCLUSIONS: It was found that field practices and winemaking processes played distinct roles in the elemental composition of grapes and wine from different preparation stages. Moreover, PIXE proved to be a suitable analytical tool for the analysis of the whole wine cycle because it handles solid and liquid samples in a clean and straightforward manner. © 2019 Society of Chemical Industry.

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Tobacco farming has been proving to induce poor health outcomes in agricultural workers, genomic instability being the triggering one. This study evaluated influence of PON1 (paraoxonase 1), SOD2 (superoxide dismutase), OGG1 (8-oxoguanine glycosylase), XRCC1 (X-ray repair cross-complementing protein 1), and XRCC4 (X-ray repair cross-complementing protein 4) genes polymorphisms on DNA damage in 121 subjects occupationally exposed to pesticides mixtures and nicotine at tobacco fields and 121 non-exposed individuals. Inorganic elements (Cl, P, S and Zn) and cotinine levels were found increased in farmers, confirming exposure. Results show higher frequencies of buccal micronucleus (MN), nuclear buds (NBUD), binucleated cells (BN) and damage index (comet assay), reduced telomere length (TL), and increased parameters of oxidative stress in farmers compared to non-exposed individuals. PON1 Gln/Gln genotype was associated with increased MN frequency. SOD2 Val/Val showed association with increased frequency of MN and NBUD and decreased antioxidant activity. The XRCC1 Arg/Arg showed protective effect for MN, BN and TL, which was also positively influenced by OGG1 -/Cys. MN was decreased in XRCC4 -/Ile farmers. These genotypes also showed a risk for antioxidant activity. Our study proposes that PON1 and SOD2 variants play a role in xenobiotic-metabolizing system in farmers, while base excision repair (BER) pathway could be the repair mechanism involved in genomic instability suffered by tobacco farmers.

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Tobacco farming is an important economic income in Brazil, although it has been challenged as regard the occupational exposure to both pesticides and nicotine endured by farmers. Chronic occupational exposure to complex mixtures can lead to health hazardous. We examined genomic instability and epigenetic changes in tobacco farmers occupationally exposed to pesticide mixtures and nicotine at tobacco fields. DNA damage was assessed by alkaline comet assay in blood cells. Genomic DNA was isolated, and telomere length was measured using quantitative polymerase chain reaction assay. We measured 5-methyl-2'-deoxycytidine, a marker of global DNA methylation, and p16 promoter methylation. The oxidative profile was evaluated by trolox equivalent antioxidant capacity and lipid peroxidation (thiobarbituric acid reactive substances) in serum. Exposure parameters, plasma cotinine and inorganic element levels, were also measured. DNA damage was significantly elevated for farmers in relation to unexposed group (P < 0.001; Mann-Whitney test) and positively associated with years of exposure. Inverse relationship between DNA damage and total equivalent antioxidant activity was demonstrated for exposed and unexposed groups. Exposed group showed significantly shorter telomeres (P < 0.001; unpaired t-test) and DNA hypomethylation (P < 0.001; unpaired t-test), as well as p16 hypermethylation (P = 0.003; Mann-Whitney test). Lipid peroxidation was increased for exposed group in relation to unexposed one (P = 0.02; Mann-Whitney test) and presented a positive correlation with global DNA methylation (P = 0.0264). Farmers have increased plasma cotinine levels (P < 0.001) and inorganic elements (phosphorus, sulphur and chlorine) in relation to unexposed group. Elevated oxidative stress levels due to chronic occupational pesticide mixtures and nicotine exposure in tobacco farmers were associated with higher DNA damage, shorter telomeres and altered DNA methylation. Telomere-accelerated attrition due to exposure may be potential intermediate step before a disease state.

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Epidemiological studies indicate that living in proximity to coal mines is correlated with numerous diseases including cancer, and that exposure to PM10 and PM2.5 components could be associated with this phenomenon. However, the understanding of the mechanisms by which PM exerts its adverse effects is still incomplete and comes mainly from studies in occupationally exposed populations. The aims of this study were to: (1) evaluate DNA damage in lymphocytes assessing the cytokinesis-block micronucleus cytome assay (CBMN-cyt) parameters; (2) identify aneugenic or clastogenic effects in lymphocytes of exposed populations using CREST immunostaining for micronuclei; (3) evaluate multi-elemental composition of atmospheric particulate matter; and (4) verify relation between the DNA damage and PM2.5 and PM10 levels around the mining area. Analysis revealed a significant increase in micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells of individuals with residential proximity to open-pit coal mines compared to residents from non-mining areas. Correlation analysis demonstrated a highly significant association between PM2.5 levels, MNBN frequencies and CREST+ micronuclei induction in exposed residents. These results suggest that PM2.5 fraction generated in coal mining activities may induce whole chromosome loss (aneuploidy) preferentially, although there are also chromosome breaks. Analysis of the chemical composition of PM2.5 by PIXE demonstrated that Si, S, K and Cr concentrations varied significantly between coal mining and reference areas. Enrichment factor values (EF) showed that S, Cr and Cu were highly enriched in the coal mining areas. Compared to reference area, mining regions had also higher concentrations of extractable organic matter (EOM) related to nonpolar and polar compounds. Our results demonstrate that PM2.5 fraction represents the most important health risk for residents living near open-pit mines, underscoring the need for incorporation of ambient air standards based on PM2.5 measures in coal mining areas.

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We assessed elemental composition of the liver in mice subjected to one-time or chronic consumption of the juice of vegetables cultivated in a vegetable garden built over deposits of coal waste. Lactuca sativa L. (lettuce), Beta vulgaris L. (beet), Brassica oleracea L. var. italica (broccoli) and Brassica oleracea L. var. acephala (kale) were collected from the coal-mining area and from a certified organic farm (control). Elemental composition was analyzed by particle-induced X-ray emission (PIXE) method. Concentrations of Mg, S, and Ca of mice subjected to one-time consumption of broccoli and concentrations of these same elements plus Si of mice receiving kale were higher in the coal-mining area. Concentrations of P, K, and Cu were increase after chronic consumption of lettuce from the coal-mining area, whereas the levels of Si, P, K, Fe, and Zn were higher in the group consuming kale from the coal-mining area. Our data suggests that people consuming vegetables grown over coal wastes may ingest significant amounts of chemical elements that pose a risk to health, since these plants contain both essential and toxic metals in a wide range of concentrations, which can do more harm than good.

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ABSTRACT We assessed elemental composition of the liver in mice subjected to one-time or chronic consumption of the juice of vegetables cultivated in a vegetable garden built over deposits of coal waste. Lactuca sativa L. (lettuce), Beta vulgaris L. (beet), Brassica oleracea L. var. italica (broccoli) and Brassica oleracea L. var. acephala (kale) were collected from the coal-mining area and from a certified organic farm (control). Elemental composition was analyzed by particle-induced X-ray emission (PIXE) method. Concentrations of Mg, S, and Ca of mice subjected to one-time consumption of broccoli and concentrations of these same elements plus Si of mice receiving kale were higher in the coal-mining area. Concentrations of P, K, and Cu were increase after chronic consumption of lettuce from the coal-mining area, whereas the levels of Si, P, K, Fe, and Zn were higher in the group consuming kale from the coal-mining area. Our data suggests that people consuming vegetables grown over coal wastes may ingest significant amounts of chemical elements that pose a risk to health, since these plants contain both essential and toxic metals in a wide range of concentrations, which can do more harm than good.

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Pollution of aquatic ecosystems is associated with the discharge of mostly industrial and urban effluents, which may cause loss of biodiversity and damage to public health. This study aims to evaluate the toxicity and mutagenicity of water samples collected in the Corrente River, a major waterway in the river basin district of Pedro II, Piauí (Brazil). This river is exposed to intense anthropogenic influence from urban, automotive mechanical and family farm waste, and it is used as the main source of water supply by the population. Water samples were collected during the rainy and dry seasons, at four sites in the Corrente River, and evaluated by physicochemical, microbiological and inorganic elements analyses. The samples were evaluated for mutagenicity using the Allium cepa test (toxicity, chromosomal aberration and micronucleus tests) and fish (Tilapia rendalli and Hoplias malabaricus). The physicochemical, microbiological and inorganic results show a large contribution to the pollution loads at collection points in the town of Pedro II, demonstrating the influence of urban pollution. The Al, Si, Ti, Cr, Ni and Cu contents were determined by PIXE. These same Corrente River water samples demonstrated mutagenic effect for A. cepa and fish, as well as toxicity in the A. cepa test. The observations of mutagenic effect may suggest that the complex mixture of agents is comprised of both clastogenic and aneugenic agents. This study also showed the need for constant monitoring in places with environmental degradation caused by urban sewage discharges.

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Background. Industrial processing of the pecan nut Carya illinoinensis K. Koch generated a large amount of shells, which have been used to prepare nutritional supplements and medicinal products; however, the safe use of shells requires assessment. This study evaluated the toxic, genotoxic, and mutagenic effects of pecan shell aqueous extract (PSAE) and the possible contribution of phenolic compounds, ellagic and gallic acids, and inorganic elements present in PSAE to induce toxicity. Results. Levels of inorganic elements like K, P, Cl, and Rb quantified using the Particle-Induced X-Ray Emission method were higher in PSAE than in pecan shells, while Mg and Mn levels were higher in shells. Mice showed neurobehavioral toxicity when given high PSAE doses (200-2,000 mg kg(-1)). The LD50 was 1,166.3 mg kg(-1). However, PSAE (50-200 mg·kg(-1)) and the phenolic compounds (10-100 mg·kg(-1)) did not induce DNA damage or mutagenicity evaluated using the comet assay and micronucleus test. Treatment with ellagic acid (10-100 mg·kg(-1)) decreased triglyceride and glucose levels, while treatments with PSAE and gallic acid had no effect. Conclusion. Pecan shell toxicity might be associated with high concentrations of inorganic elements such as Mn, Al, Cu, and Fe acting on the central nervous system, besides phytochemical components, suggesting that the definition of the safe dose should take into account the consumption of micronutrients.

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Agricultural workers involved in the harvest of tobacco crops are regularly exposed to large quantities of pesticides. In order to determine how this exposure to pesticides induces genetic alterations in these workers, blood samples were obtained from 77 exposed individuals, as well as from 60 unexposed subjects. DNA damage was analyzed by the Comet assay and by the micronucleus (MN) test. The antioxidant profile was evaluated by activity of superoxide dismutase (SOD), and the polymorphism of gene PON1 was used as a susceptibility biomarker. The content of inorganic elements in the blood samples was determined by PIXE analysis. Our results demonstrated that the damage frequency, damage index, the MN frequency, and the SOD activity were significantly elevated in the exposed relative to the unexposed group. A modulation of the MN results for the PON1 gene was observed in the exposed group. The concentrations of inorganic elements in the exposed group were higher compared to those of the unexposed group. In this study, we observed that genetic damage, and change in oxidative balance were induced by the exposure of workers to complex mixtures of pesticides in the presence of inorganic compounds, whereby an influence of the genotype was evident.

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The entire process of power generation, extraction, processing and use of coal strongly impact water resources, soil, air quality and biota leads to changes in the fauna and flora. Pollutants generated by coal burning have been contaminating plants that grow in area impacted by airborne pollution with high metal contents. Baccharis trimera is popularly consumed as tea, and is widely developed in Candiota (Brazil), one of the most important coal burning regions of the Brazil. This study aims to investigate the phytochemical profile, in vivo genotoxic and mutagenic potential of extracts of B. trimera collected from an exposed region to pollutants generated by coal burning (Candiota City) and other unexposed region (Bagé City), using the Comet assay and micronucleus test in mice and the Salmonella/microsome short-term assay. The HPLC analyses indicated higher levels of flavonoids and phenolic acids for B. trimera aqueous extract from Bagé and absence of polycyclic aromatic hydrocarbons for both extracts. The presence of toxic elements such as cobalt, nickel and manganese was statistically superior in the extract from Candiota. For the Comet assay and micronucleus test, the mice were treated with Candiota and Bagé B. trimera aqueous extracts (500-2000 mg/kg). Significant genotoxicity was observed at higher doses treated with B. trimera aqueous extract from Candiota in liver and peripheral blood cells. Micronuclei were not observed but the results of the Salmonella/microsome short-term assay showed a significant increase in TA98 revertants for B. trimera aqueous extract from Candiota. The extract of B. trimera from Candiota bioacumulated higher levels of trace elements which were associated with the genotoxic effects detected in liver and peripheral blood cells.

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SCOPE: Postmenopausal women are often affected by a group of metabolic disorders related to oxidative stress. Alternative treatments that can improve the quality of life of these women have been the subject of recent studies. The objective of this study was to evaluate the response to oxidative stress in the brains of rats following ovariectomy, and to determine enzymatic and nonenzymatic antioxidant responses when the animals received 3 months of dietary supplementation. METHODS AND RESULTS: Ovariectomy produced changes in antioxidant profiles characterized by reductions in glutathione S-transferase activity, H2 O2 consumption, superoxide dismutase activity, and vitamin C levels and increases in protein carbonylation. Docosahexaenoic fatty acid (DHA) supplementation restored these parameters to normal values and increased values of other antioxidants (glutathione peroxidase and total glutathione). However, DHA supplementation also increased protein carbonylation and lipid peroxidation. Eicosapentaenoic acid supplementation produced no changes in antioxidants, but decreased lipid peroxidation. Lipoic acid supplementation increased consumption of H2 O2 and decreased protein carbonylation and lipid peroxidation. CONCLUSION: These results suggest that the antioxidant response to omega-3 varies in different tissues, and in this study DHA treatment had a prooxidant effect in the brain. Lipoic acid treatment, on the other hand, had a protective effect, reducing markers of oxidative damage.

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During coal combustion, hazardous elements are discharged that impair environmental quality. Plant cover is the first available surface for the atmospheric pollutants in terrestrial ecosystems. The aim of this study was to evaluate genotoxicity in the aqueous extract of the native plant, Baccharis trimera, exposed to coal and emissions from a thermal power plant (coal-fired power plant in Candiota, Brazil), correlating seasonality, wind tunnel predominance, and presence of inorganic elements. The presence of inorganic elements in the aerial parts of B. trimera was analyzed by particle-induced X-ray emission (PIXE) spectrometry, and genotoxicity was evaluated by ex vivo comet assay. The genotoxic effects of aqueous extracts of B. trimera from four sites located in the area around power plant were analyzed by comet assay in peripheral human lymphocytes. Winter samples showed greater levels of metals than summer samples. Genotoxicity was detected in B. trimera extracts collected from the region exposed to extraction and burning coal. Extracts from the site impacted by the dominant wind induced more damage to DNA than those from other sites. Based on our data, we can suggest that in winter the inorganic elements from extraction and burning of coal and carried through the wind tunnel were responsible for the genotoxicity observed in aqueous extract of B. trimera.

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Coal is the largest fossil fuel source used for the generation of energy. However, coal extraction and its use constitute important pollution factors; thus, risk characterization and estimation are extremely important for the safety of coal workers and the environment. Candiota is located to the southeast of the state of Rio Grande do Sul and has the largest coal reserves in Brazil, and the largest thermal power complex in the state. In the open-cast mines, the coal miners are constantly exposed to coal dust. The human buccal micronucleus cytome (BMCyt) assay has been used widely to investigate biomarkers for DNA damage, cell death, and basal cell frequency in buccal cells. The aim of this study was to assess whether prolonged exposure to coal dust could lead to an increase in genomic instability, cell death, and frequency of basal cells using the BMCyt assay. In the analysis of epithelial cells, the exposed group (n = 41) presented with a significantly higher frequency of basal cells, micronuclei in basal and differentiated cells, and binucleated cells compared to the non-exposed group (n = 29). The exposed group showed a significantly lower frequency of condensed chromatin cells than the non-exposed group. However, we found no correlation between DNA damage and metal concentration in the blood of mine workers. DNA damage observed in the mine workers may be a consequence of oxidative damage resulting from exposure to coal residue mixtures. In addition, our findings confirm that the BMCyt assay can be used to identify occupational risk.

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Tobacco farming presents several hazards to those who cultivate and harvest the plant. The genotoxic and mutagenic effects in tobacco farmers were investigated. In order to verify the relationship between genetic susceptibility and biomarkers GSTT1, GSTM1, GSTP1, CYP2A6, PON, OGG1, RAD51, XRCC1, and XRCC4 genes polymorphism were evaluated. Oxidative stress markers and trace elements content were determined. Peripheral blood cells samples were collected from 111 agricultural workers during pesticides application and leaf harvest, and 56 non-exposed subjects. Results show that farmers are exposed to mixture of substances with genotoxic and cytotoxic potential. Only GSTM1 null and CYP2A6*9 showed significant associations with cytokinesis-blocked micronuclei assay results. In pesticide application an increase in trace elements content was observed. The results indicated that exposure to pesticides and nicotine can influence antioxidant enzymes activity. Our study drives the attention once more to the need for occupational training on safe work environment for farm workers.

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Fruits are rich in minerals, which are essential for a wide variety of metabolic and physiologic processes in the human body. The use of frozen fruits has greatly spread in the last years not only in the preparation of juices, but also as raw material for yogurts, candies, cookies, cakes, ice creams, and children's food. However, up to now there is no data about the mineral profile of frozen fruits. This is the first database to quantify the levels of minerals in 23 samples of frozen fruits, including the most used around the world and some native fruits from the Amazon rainforest in Brazil. Considering the Dietary Reference Intakes, 100g of frozen fruits can provide 0.2 to 2.8% of macro and 2.5 to 100% of microminerals for adults (31-50 years old). Although geographical differences should be considered, these data can help to plan diets and to develop population interventions aiming to prevent chronic diseases.

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Fruits are rich in minerals, which are essential for a wide variety of metabolic and physiologic processes in the human body. The use of frozen fruits has greatly spread in the last years not only in the preparation of juices, but also as raw material for yogurts, candies, cookies, cakes, ice creams, and children's food. However, up to now there is no data about the mineral profile of frozen fruits. This is the first database to quantify the levels of minerals in 23 samples of frozen fruits, including the most used around the world and some native fruits from the Amazon rainforest in Brazil. Considering the Dietary Reference Intakes, 100g of frozen fruits can provide 0.2 to 2.8 percent of macro and 2.5 to 100 percent of microminerals for adults (31-50 years old). Although geographical differences should be considered, these data can help to plan diets and to develop population interventions aiming to prevent chronic diseases.

As frutas são ricas em minerais, sendo estes essenciais para uma grande variedade de processos metabólicos e fisiológicos no corpo humano. A utilização de frutas congeladas tem se ampliado nos últimos anos, não só na preparação de sucos, mas também como matéria-prima para iogurtes, doces, biscoitos, bolos, sorvetes e alimentos infantis. No entanto, até o momento não há dados sobre o perfil mineral de frutas congeladas. Este trabalho é o primeiro banco de dados para quantificar os níveis de minerais em 23 amostras de frutas congeladas, bastante consumidas em todo o mundo e de algumas frutas nativas da floresta amazônica, Brasil. Considerando-se as Referências de Ingestão Diárias, 100g de frutas congeladas podem fornecer 0,2-2,8 por cento de macro e de 2,5 a 100 por cento dos mi-crominerais para adultos (31-50 anos). Embora as diferenças geográficas devam ser consideradas, estes dados ajudam para o plano de dietas e desenvolvimento de intervenções junto à população co o objetivo de prevenir doenças crônicas.

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There are many studies related to the antioxidant activity of grape products; however, they concern only purple and white grape varieties. Up to now, there are no reports of studies on the Goethe rose grape variety, either on its antioxidant activity or on its phenolic and mineral quantification. Thus, the purpose of this study was to evaluate in vitro and in vivo antioxidant activity, as well as to quantify total phenolic compounds, ascorbic acid, and mineral content, in a Goethe rose grape juice. The results obtained showed that the Goethe rose grape juice is a great polyphenol source, which contains catechin, epicatechin, and procyanidins (B(1), B(2), B(3), and B(4)). Of all metals analyzed, potassium, calcium, magnesium, and iron showed the highest values. We found that this rose grape juice shows an important antioxidant activity in in vitro (2,2-diphenyl-1-picrylhydrazyl radical scavenging activity) and in vivo (using the Saccharomyces cerevisiae yeast cells) assays. The antioxidant activity could be explained by the significant phenolic content and ascorbic acid levels found in the juice. The results showed that rose grape juice is an excellent antioxidant source, which could contribute to the prevention of many diseases related to oxidative stress, such as atherosclerosis and Parkinson's disease.

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Industrial effluents, agricultural runoff, and municipal wastewaters contain unknown substances and complex mixtures that are released into the environment and can lead to contamination of surface and subsurface waters. In the present report, we have used the alkaline Comet assay and the micronucleus (MN) test to detect the genotoxicity due to multiple sources of pollution in the peripheral blood of two native estuarine fish (mullet and sea catfish) and evaluated possible interactive genotoxic effects from multiple contaminants and the seasonal variation of the genotoxicity. Mullet and sea catfish were captured in the Tramandai and Mampituba Rivers in the southern Brazilian state of Rio Grande do Sul. Reference animals were obtained from the Armazem lagoon. Fish captured in the two estuaries during the four seasons over a period of 2 years had increased levels of DNA damage and MN frequencies relative to the reference fish. In general, the alkaline Comet assay was more sensitive to the genotoxicity of the river contaminants than the MN test. The Comet assay demonstrated significant differences in fish captured at different seasons and at the two river sites, while the MN test showed significant differences only for the annual average for mullet from both sites and fish from the control site. The increases in DNA damage appear to be related to the increase in the number of people in the towns close to the study areas during the warm spring and summer seasons. Although no specific cause-effect relationships were established, comparison of the chemical contaminants and physical variations in the rivers with the genotoxicity data indicate that there may be some association between hydrocarbons, metals, pH, and water temperature and the level of damaged cells observed in mullet and sea catfish from the Tramandai and Mampituba estuaries.

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