The use of an alimentary index to assess anthropogenic debris on green turtles (Chelonia mydas).

Studies on novel methodologies addressed to quantification of anthropogenic marine debris (AMD) types ingested by green turtles (Chelonia mydas) is not common in the scientific literature. To fill this gap, we used the alimentary index (IAi) to quantify and classify AMD ingested by green turtles stranded in the Potiguar Basin, northeastern Brazil. We classified 295 green turtles as juveniles (JUV-I (n = 190) and JUV-II (n = 58)) and adults (ADU (n = 47)). We collected gut contents during necropsy and analyzed them using the IAi. The IAi values were categorized as main (IAi ≥ 0.5), secondary (0.25 ≤ IAi < 0.5), and accessory (IAi < 0.25). The results revealed that transparent soft plastic was an accessory item for the three size groups and the second most frequent item (IAi = 0.23) for JUV-I. Our study brings insights into the application of the IAi to evaluate the amount and types of AMD ingested by green turtles.

The link between pharmaceuticals and cyanobacteria: a review regarding ecotoxicological, ecological, and sanitary aspects.

Cyanobacteria are important for ecosystem functioning, but eutrophication may affect the surrounding biome by losing ecosystem services and/or through affecting the cyanotoxins production that threatens ecological and human health. Pollution is an environmental issue that affects aquatic ecosystems worldwide, and the knowledge of the role of synthetic chemicals such as pharmaceuticals is still scarce. Therefore, studies coupling these two relevant issues are essential to better understand the ecological risks and the potential threats to public health. Thus, an overview of ecotoxicological tests performed in the literature exposing cyanobacteria to pharmaceuticals and the possible consequences regarding ecological and sanitary aspects was conducted. Moreover, a risk assessment was performed to enable a better understanding of pharmaceuticals affecting cyanobacteria ecology. Most of the studies found in the literature tested isolated pharmaceuticals in laboratory conditions, while others assessed mixture effects on in situ conditions. The endpoints most assessed were growth, photosynthesis, and antioxidant enzyme activity. The studies also point out that cyanobacteria may present resistance or sensitivity depending on the concentrations and the therapeutic class, which may cause a change in the ecosystem dynamics and/or sanitary implications due to cyanotoxin production. The risk assessment highlighted that antibiotics are among the most relevant substances due to the chemical diversity and higher levels found in the environment than other therapeutic classes. This review highlighted gaps regarding cyanotoxin release into aquatic environments due to the occurrence of pharmaceuticals and the need for more realistic experiments to better understand the potential consequences for human and environmental health.

Natural radioactivity in Brazil: a systematic review.

Natural radioactivity is a public health concern worldwide. Its deleterious effects are largely associated with emitting ionizing particles which generate innumerable toxicological consequences to human being. The present study aimed to describe the research state of the art on natural radioactivity in Brazil through a systematic review limited to articles published in the twenty-first century in the PubMed, SciELO, Lilacs, and Google Scholar databases. A total of 55 research articles were considered for this purpose. Based on the collected sample types, the radiation analysis in most of the scientific reports was performed on solid samples (soil/sediment/rocks), followed by water and air. In fact, most of the available information came from geological studies. A wide range of concentrations and a variety of radionuclides have been assessed, with radium being the most cited. Most of the studies described radiation levels above the international guidelines, and consider the Brazilian territory as a high natural background radiation region (HNBR). In comparison with other HNBR areas, the scientific information about the related risks to human health is still scarce. There is uncertainty about the real impact of natural radioactivity on human health, as there is a lack of scientific information for most of the country about this issue. The analysis and comparison of the available information highlights the potential risks linked to natural radioactivity and the need to incorporate suitable environmental management policies about this issue.

Toxicological aspects of trihalomethanes: a systematic review.

Chlorine is considered the most used chemical agent for water disinfection worldwide. However, water chlorination can lead to by-product generation which can be toxic to humans. The present study aimed to perform a systematic review on the toxicity of trihalomethanes (THMs) through bioindicators of cytotoxicity, genotoxicity, and mutagenicity. The results showed that studies on the effects of THMs on DNA are a current research concern for evaluating the toxicity of the pure compounds and real samples involving several types including water for recreational use, reused water, and drinking water. THMs deleterious effects have been assessed using several biosystems, where the Ames test along with experimental animal models were the most cited. A wide range of THM concentrations have been tested. Nevertheless, DNA damage was demonstrated, highlighting the potential human health risk. Among the studied THMs, chloroform presented a different action mechanism when compared with brominated THMs, with the former being cytotoxic while brominated THMs (bromodichloromethane, bromoform, and dibromochloromethane) were cytotoxic, genotoxic, and mutagenic. The described evidence in this research highlights the relevance of this topic as a human health issue. Nevertheless, research aimed to represent THMs current exposure conditions in a more accurate way would be needed to understand the real impact on human health.

Diosgenin induces genotoxic and mutagenic effects on HepG2 cells.

Yam roots and other plants from Dioscorea genus have cultural, nutritional and economic importance to tropical and subtropical regions and have a great amount of diosgenin in its composition. In the present study the cytotoxic, genotoxic and mutagenic potential of diosgenin on HepG2 cells was investigated. Cytotoxicity was assessed using MTT and clonogenic assay. Genotoxic and mutagenic effects were performed using single cell gel electrophoresis and cytokinesis-block micronucleus assay, respectively. A reduction on cell viability was observed due to diosgenin treatment at concentrations higher than 30 µM. A genotoxic effect was shown by comet assay and CBMN. Besides, an increase in micronucleus frequency along with a significant cytostatic effect were observed. Diosgenin elicited DNA damage on HepG2 cells which could not be efficiently repaired contributing to the mutagenic effect observed. Those results suggest that diosgenin deleterious effect could take place through genetic instability, fact that affects the normal cell cycle, leading to cell's death.

Genotoxicity of Turnera subulata and Spondias mombin × Spondias tuberosa Extracts from Brazilian Caatinga Biome.

Medicinal plants have been used in primary healthcare since the earliest days of humankind. Turnera subulata and Spondias mombin × Spondias tuberosa are widely used in the Brazilian Northeast to treat several diseases. The aim of this study was to evaluate the genotoxic effects of the leaf extracts of these species by the somatic mutation and recombination test in the somatic cells of Drosophila melanogaster wings. The experiments were performed using standard and high-bioactivation cross and three concentrations of the test substance [aqueous extract (AET and AES) at 5.0, 10.0, and 20.0 mg/mL and ethanolic extract (EET and EES) and ethyl acetate fraction (EAFT and EAFS) at 0.625, 1.25, and 2.5 mg/mL]. Results indicated that the extracts and fractions induced spontaneous frequencies of mutant spots in both D. melanogaster crosses. Nevertheless, the highest concentrations of the tested plant chemical agents were responsible for the statistically significant genotypic effect. T. subulata and S. mombin × S. tuberosa displayed genotoxic effect under the experimental conditions. The results from this study are crucial as they indicated the deleterious and side effects, considering the indiscriminate use of the extracts of these plants for disease treatment.

The use of bacterial bioremediation of metals in aquatic environments in the twenty-first century: a systematic review.

Metal pollution is a current environmental issue as a consequence of unregulated anthropic activiy. A wide range of bioremediation strategies have been successfully implemented to recover contaminated areas. Among them, bacterial bioremediation stands out as a promising tool to confront these types of concerns. This study aimed to compare and discuss worldwide scientific evolution of bacterial potential for metal bioremediation in aquatic ecosystems. The study consisted of a systematic review, elaborated through a conceptual hypothesis model, during the period from 2000 to 2016, using PubMed, MEDLINE, and SciELO databases as data resources. The countries with the largest number of reports included in this work were India and the USA. Industrial wastewater discharge was the main subject associated to metal contamination/pollution and where bacterial bioremediations have mostly been applied. Biosorption is the main bioremediation mechanism described. Bacterial adaptation to metal presence was discussed in all the selected studies, and chromium was the most researched bioremedied substrate. Gram-negative Pseudomonas aeruginosas and the Gram-positive Bacillus subtilis bacteria were microorganisms with the greatest applicability for metal bioremediation. Most reports involved the study of genes and/or proteins related to metal metabolism and/or resistence, and Chromobacterium violaceum was the most studied. The present work shows the relevance of metal bacterial bioremediation through the high number of studies aimed at understanding the microbiological mechanisms involved. Moreover, the developed processes applied in removal and/or reducing the resulting environmental metal contaminant/pollutant load have become a current and increasingly biotechnological issue for recovering impacted areas.

Analysis of genome instability biomarkers in children with non-syndromic orofacial clefts.

The non-syndromic cleft lip and/or palate (NSCL/P) is a common birth defect caused by a combination of genetic and environmental factors. The possible role of genome instability on NSCL/P patient needs more investigation, since DNA metabolism is an essential cellular function to keep cells with normal genotypes and gene expression patterns according to tissue specificities, which is critical during embryo development because it requires sensitive regulation of cell proliferation, apoptosis and differentiation. Thus, genome stability is ultimately essential to maintain a healthy life. The aim of this study was to assess the frequency of genome instability biomarkers and their relationship with NSCL/P. Cytokinesis-block micronucleus assay was performed to estimate the biomarkers frequency and gene expression was analyzed by the transcriptogram in order to further explore the role of genome instability and other biological processes in this birth defect. The NSCL/P patients had higher baseline frequency of micronucleus, nuclear buds and nucleoplasmic bridges (P < 0.001) than the control group. Moreover, new nuclear morphologies (fused, circular and horseshoe) was detected in the patients' cells analyzed, possibly indicating that chronic folic acid deficiency is interfering in their genome instability. Children with clefts had 2.3 times more risk to have high micronuclei frequency (P = 0.043) according to binary logistic regression. The high genomic instability in children with oral clefts suggests that misrepaired double strand breaks in DNA that create micronuclei representing a significant factor in NSCL/P development. This study was published in 52nd EUROTOX Abstract Book.

Mutagenic potential assessment associated with human exposure to natural radioactivity.

Lucrécia city, known to harbor a high cancer rate, is located in a semiarid region characterized by the presence of mineral reservoirs, facing a high exposure to metal and natural radioactivity. The present study aimed to assess the environmental scenario at a semiarid region located in Northeastern Brazil. Metal concentration, alpha and beta radiation, and cyanobacteria content in tap water along with indoor radon and gamma emitters (U, K and Th) concentrations were measured. In addition, mutagenic and nuclear instability effects were assessed using buccal micronucleus cytome assay. The study included five samplings corresponding to a period between 2007 and 2009. Drinking water from Lucrécia city presented levels of Mn, Ni and Cr along with cyanobacteria in concentrations one to four times higher than regulatory guidelines considered. Furthermore, high levels of all the tested radionuclides were found. A high percentage of the houses included in this study presented indoor radon concentrations over 100 Bq m-3. The mean annual effective dose from Lucrécia houses was six times higher than observed in a control region. The levels of exposure in most of the Lucrécia houses were classified as middle to high. A significant mutagenic effect, represented as an increase of micronuclei (MN) frequency and nuclear abnormalities as nuclear buds (NB), binucleated cells (BN), and pyknotic cells (PYC) were found. The results obtained highlight the role of high background radioactivity on the observed mutagenic effect and could help to explain the exacerbated cancer rate reported in this locality.

Ecotoxicological water assessment of an estuarine river from the Brazilian Northeast, potentially affected by industrial wastewater discharge.

Water pollution generated by industrial effluents discharge is a threat to the maintenance of aquatic ecosystems and human development. The Jundiai River estuarine, located in Northeast Brazil, receives an industrial pretreated effluent load from the city of Macaíba/RN/Brazil. The present study aimed to assess the water quality of this water reservoir through i) physicochemical characterization, ii) quantification of metal concentration and iii) by an ecotoxicological assessment carried out using Mysidopsis juniae and Pomacea lineata. The study was performed throughout the period comprising May to September 2014. Physicochemical variables such as chloride, total solids and electrical conductivity presented values in the waste discharge point, significantly different with those located out of the waste releasing point. Apart from that, metal concentration showed variable behavior throughout the monitored period. Levels of Al, Fe, Cu, Cd, Cr, Ni, Pb and Ag were over the considered guidelines. Both natural and anthropogenic sources seem to be involved in the resulting environmental scenario. A reduction in the fecundity rate (using Mysidopsis juniae) along with an increase in mortality rate (in both species) was observed ratifying the presence of toxic substances in this water reservoir. Moreover, a correlation analysis stated an association of the aforementioned toxicological effects with the delivery of industrial waste products. The ecotoxicological assessment performed highlighted the presence of toxic substance/s in water from the Jundiai River. Especially as a consequence of industrial activity, a fact that might threaten the bioma and, therefore, the human health of the population settled in the studied region.

Micronucleus study of the quality and mutagenicity of surface water from a semi-arid region.

The present study evaluated the mutagenic potential of surface water from the Lucrecia dam. The Tradescantia-micronucleus (Trad-MCN) test and CBMN assay in human peripheral blood lymphocytes were applied, corresponding to an in vivo and in vitro system, respectively. Heavy metals and some physicochemical properties were also measured. Water samples were collected in November 2009 (dry season) and May 2010 (rainy season) at three different points. Results of both assays for raw water showed positive responses for the points analyzed when compared to the negative control. The CBMN assay showed that diluted water was still able to induce a significant increase in micronucleus frequency. For both assays, the highest mean MN was observed in the dry season. Chemical analyses detected an increase in heavy metal levels at the sampling points and in the different seasons. These findings indicate the presence of genotoxins, such as heavy metals, in the water, which may be affecting the entire ecosystem, as well as human health. More prolonged monitoring is recommended in order to better characterize this public water supply.

Use of native species Crenicichla menezesi (Ariidae) as a model for in situ evaluation of genotoxicity in surface water.

The aim of this work was to analyze the genotoxicity in surface water from the Pitimbú River (Natal, Brazil) using a neotropical species Crenicichla menezesi as bioindicator for in situ evaluation. The genotoxicity was analyzed using the micronucleus (MN) and comet assays in erythrocytes. The animals were collected from five sites of the river. A significant increase of MN frequency and comet class L1 to L4 were observed only in specimens collected from three sampling sites (SS3, SS4 and SS5) located into urban areas. These same sites also presented high metals concentration. Correlation of DNA damage and concentrations of Cu (Spearman r=0.8080), Cr (Spearman r=0.9772), and Ni (Spearman r=0.9909) were observed. These data indicate the presence of genotoxic agents. Moreover the C. menezesi species showed to be a good indicator for prompt analyses since this species presents sedentary and carnivore habits.

Genotoxic analysis in aquatic environment under influence of cyanobacteria, metal and radioactivity.

The micronucleus (MN) and nuclear abnormality (NA) tests were employed to evaluate the genotoxic potential of the Lucrecia dam (RN, Brazil) located in a semi-arid region and influenced by crop irrigation and irregular rainfall. The analyses of these water samples demonstrated the presence of several cyanobacteria as well as metals and radioactivity. The Nile Tilapia (Oreochromis niloticus) was used for micronucleus and nuclear abnormality assays and analysis of metal concentrations in gills and liver samples. Genotoxic assays in source water fish showed a significant increase in the frequency of nuclear damage (MN and NA) in relation to the negative control group. In conclusion, the results obtained with water surface and biological samples from the Lucrecia dam suggest that this important water resource contains a number of chemical and microbiological pollutants with genotoxic potential and that these substances may be compromising the species inhabiting this ecosystem. Particular care must be taken to prevent further degradation of this water supply. Capsule: A battery of assay was successfully applied to assess the water quality of Lucrecia dam from a Northeastern region of Brazil.

The evaluation of maté (Ilex paraguariensis) genetic toxicity in human lymphocytes by the cytokinesis-block in the micronucleus assay.

The present study evaluates the clastogenic and/or aneugenic potential of maté (Ilex paraguariensis) - previously tested for the presence of 48 organophosphorous pesticides - in the culture of human lymphocytes in the absence of exogenous metabolic activation. Peripheral blood was obtained once from three healthy female donors for lymphocyte cell cultures. The cultures were treated with maté infusion (filtered in sterilized sartorius filter with a 0.22 mm pore membrane), distilled water (negative control), and 6 microg/ml bleomycin (positive control). For each experimental person, 3000 binucleated cells (BN) from two independent cultures (1000 cells from replicate cultures) were scored for the presence of micronuclei (MN). No statistical differences between maté infusion concentrations were observed: 1400 microg/ml (0.001+/-0.002), 700 microg/ml (0.0006+/-0.0015), 350 microg/ml (0.002+/-0.002), 175 microg/ml (0.002+/-0.003) and negative control (0.001+/-0.001). The present findings show that there is no clastogenic or/and aneugenic basis underlying maté action in the CBMN assay.

Mutagenic and recombinagenic activity of airborne particulates, PM10 and TSP, organic extracts in the Drosophila wing-spot test.

The genotoxicity associated with air pollution in the city of Canoas, Rio Grande do Sul (Brazil), was assessed in November (spring) and January (summer). We applied the somatic mutation and recombination test (SMART) in Drosophila melanogaster in its standard version with normal bioactivation (ST) and in its variant with increased cytochrome P450-dependent biotransformation capacity (HB). The data indicated the genotoxicity of TSP and PM10 collected in November, in both ST and HB crosses. The genotoxic activity of the PM10 material in the spring sample was exclusively associated with the induction of mitotic recombination, whereas the TSP genetic toxicity was due to both recombinational as well as point and/or chromosomal mutation events. Considering PM10 collected in January, a positive response--100% (17.10 m3/ml) concentration--was observed in the HB cross, which was not detected in the ST cross.

Genotoxicity of three mouthwash products, Cepacol, Periogard, and Plax, in the Drosophila wing-spot test.

Antiseptic mouthwashes used in biofilm control are widely available in the marketplace, despite inconsistent data concerning their genetic and cellular toxicity. In the present study, we investigated the genotoxic potential of three antiseptics currently used for odontologic treatment, Cepacol (containing cetylpyridinium chloride), Periogard (chlorhexidine digluconate), and Plax (triclosan). Genotoxicity was evaluated using the Somatic Mutation and Recombination Test (SMART) in Drosophila melanogaster, employing flies having normal bioactivation (the standard cross) and flies with increased cytochrome P450-dependent biotransformation capacity (the high bioactivation cross). Periogard and Plax produced negative responses in both types of flies; however, Cepacol (75 and 100%) produced positive responses in both the standard and high bioactivation assays, with the genotoxic responses mainly due to the induction of mitotic recombination. Assays performed with ethanol and cetylpirydinium chloride, two major ingredients of Cepacol, indicated that the genotoxicity of the mouthwash is likely to be due to ethanol.

Vanillin as a modulator agent in SMART test: inhibition in the steps that precede N-methyl-N-nitrosourea-, N-ethyl-N-nitrosourea-, ethylmethanesulphonate- and bleomycin-genotoxicity.

Vanillin (VA), the world's major flavoring compound used in food industry and confectionery products - that has antimutagenic and anticarcinogenic activity against a variety of mutagenic/carcinogenic agents - was tested for the interval between the formation of premutational lesion and it is finalization as a DNA lesion. The overall findings using co-treatment protocols in SMART test suggest that VA can lead to a significant protection against the general genotoxicity of ethylmethanesulphonate (EMS), N-ethyl-N-nitrosourea (ENU), N-methyl-N-nitrosourea (MNU) and bleomycin sulphate (BLEO). Considering MNU, ENU and EMS the desmutagenic activity observed could result from VA-stimulation of detoxification, via induction of glutathione S-transferase. However, the protector effect related to BLEO could be attributed to its powerful scavenger ability, which has the potential to prevent oxidative damage induced by BLEO.

Genetic toxicity in surface water from Guaíba Hydrographic Region under the influence of industrial, urban and agricultural sewage in the Drosophila wing-spot test.

Mutagenic and recombinagenic activity of surface waters in the Guaíba Hydrographic Region (RS, Brazil) was investigated using the SMART in Drosophila melanogaster. Two positive results in Caí River (September 2000 and August 2001) and in Taquari River (August 2001 and February 2002)--linked to direct recombinagenic toxicants were observed. In Jacuí samples, an indirect mutagenic and recombinagenic action was detected in a September 2000 collection and a direct recombinational activity was observed in February 2002. Also in February 2002--samples from Dilúvio Brook and Guaíba Lake (GPC) were able to induce wing spots by mitotic recombinagenesis. The former sampling site showed toxicants to have a direct action, and the latter an increment in mitotic recombination that depended on metabolic action. The SMART wing test shows that all positive responses were mainly related to homologous mitotic recombination.

Drosophila wing-spot test for genotoxic assessment of pollutants in water samples from urban and industrial origin.

The Caí River (Rio Grande do Sul, Brazil) is an important watercourse that receives large amounts of industrial and untreated municipal discharges in its lower course. We employed the SMART in Drosophila melanogaster to evaluate the genotoxicity of surface waters collected from Caí sites receiving direct sewage discharge: from Montenegro (Km 52) and from São Sebastião do Caí (Km 78 and 80), and from two sites under the industrial influence (Km 13.6 and 18.6). The genotoxic analysis included three collections: March, June and September 1999, which were tested at crude sample and at 50 and 25% concentrations. Considering the industrial samples from Km 18.6 and 13.6, collected in March, June and September 1999, they were characterized as not having genetic toxicity. The urban samples collected in March--Km 52, 78 and 80--showed a significant increment in the frequencies of total spots. In Km 52 and 78 the genotoxic effect was associated to both mutational and recombinational events, although for Km 80 the increases observed were mainly related to the occurrence of homologous recombination. Moreover, the Km 80 crude sample from June and all the concentrations analyzed for Km 52 in September were also able to induce mitotic recombination. These effects were only observed in the ST cross, demonstrating the genotoxins present in the urban discharges act by direct interaction with the DNA of the somatic cells. The SMART in D. melanogaster was shown to be highly sensitive to detect genotoxic agents present in the aquatic environment, and must be better exploited for monitoring areas under anthropogenic discharges.