Another casualty of the SARS-CoV-2 pandemic-the environmental impact.

Cemetery leachate generated by the process of cadaveric decomposition is a significant contaminant of several matrices in the cemetery environment (soil, groundwater, and surface water). The biogenic amines cadaverine and putrescine stand out among the cemetery leachate contaminants, since they are potentially carcinogenic compounds. This review article presents a discussion of possible environmental impacts caused by the increase in deaths resulting from COVID-19 as its central theme. The study also aims to demonstrate the importance of considering, in this context, some climatic factors that can alter both the time of bodily decomposition and the longevity of the virus in the environment. Additionally, some evidence for the transmission of the virus to health professionals and family members after the patient's death and environmental contamination after the burial of the bodies will also be presented. Several sources were consulted, such as scientific electronic databases (NCBI), publications by government agencies (e.g., ARPEN, Brazil) and internationally recognized health and environmental agencies (e.g., WHO, OurWorldInData.org), as well as information published on reliable websites available for free (e.g., CNN) and scientific journals related to the topic. The data from this study sounds the alarm on the fact that an increase in the number of deaths from the complications of COVID-19 has generated serious environmental problems, resulting from Cemetery leachate.

PBAT biodegradable mulch films: Study of ecotoxicological impacts using Allium cepa, Lactuca sativa and HepG2/C3A cell culture.

Biodegradable mulch films are an alternative to polyethylene films used in agriculture for weed control, improving crop productivity. This change could minimize the residue production and costs related to the final disposal. Nevertheless, the environmental safety of these biodegradable products is scarcely investigated. In this work, samples of poly(butylene adipate-co-terephthalate)-PBAT mulch films, with and without UV stabilizer additives, were prepared. Aqueous extracts of soil samples, where mulch films were disposed, were investigated using bioassays with Lactuca sativa, Allium cepa, and cell culture HepG2/C3A. As PBAT is expected to suffer photodegradation and biodegradation, soil samples mixed with films before and after these processes were evaluated. Soil aqueous extracts promoted root grown (mainly hypocotyl) of L. sativa, probably due to presence of nutrients. So, to evaluate toxicity potential, in this case it was necessary to use aqueous extract prepared with soil instead of ultrapure water as the control. After doing this analysis it was observed that no adverse impacts due to PBAT films occurred. No chromosomal abnormalities were observed in A. cepa bioassay for any of tested samples. The absence of genotoxic potential was confirmed by comet assay and micronucleus test using human hepatocarcinoma cell line HepG2/C3A. These results showed that the soil did not induce damage to the tested organisms, before and after degradation of PBAT films.

Evaluation of the potential agricultural use of biostimulated sewage sludge using mammalian cell culture assays.

Among the bioremediation processes, biostimulation is an effective methodology for the decontamination of organic waste by the addition of agents that stimulate the indigenous microbiota development. Rice hull is a biostimulating agent that promotes the aeration of edaphic systems and stimulates the aerobiotic activity of soil microorganisms. The present study aimed to evaluate the efficacy of the bioremediation and biostimulation processes in reducing the toxicity of sewage sludge (SS) and to evaluate its possible application in agriculture using cytotoxic and genotoxic assays in human hepatoma cells (HepG2). SS of domestic origin was tested as both the pure product (PSS) and mixed with soil (S) and with a stimulating agent, such as rice hull (RH), in different proportions (SS + S and SS + S + RH); we also examined different remediation periods (3 months - T1 and 6 months - T2). For the PSS sample, a significant induction of micronucleus (MN) in T2 was observed with nuclear buds in all of the periods assessed, and we observed the presence of more than one alteration per cell (MN and nuclear bud) in T1 and T2. The PSS sample caused genotoxic effects in the HepG2 cells even after being bioremediated. For the samples containing soil and/or rice hull, no toxic effects were observed in the test system used. Therefore, the addition of SS to agricultural soils should be conducted with caution, and it is important that the SS undergoes a remediation process, such as bioremediation and biostimulation treatments.

Fruit extract of the medicinal plant Crataegus oxyacantha exerts genotoxic and mutagenic effects in cultured cells.

Crataegus oxyacantha, a plant of the Rosaceae family also known "English hawthorn, haw, maybush, or whitethorn," has long been used for medicinal purposes such as digestive disorders, hyperlipidemia, dyspnea, inducing diuresis, and preventing kidney stones. However, the predominant use of this plant has been to treat cardiovascular disorders. Due to a lack of studies on the genotoxicity of C. oxyacantha, this investigation was undertaken to determine whether its fruit extract exerts cytotoxic, genotoxic, or clastogenic/aneugenic effects in leukocytes and HepG2 (liver hepatocellular carcinoma) cultured human cells, or mutagenic effects in TA100 and TA98 strains of Salmonella typhimurium bacterium. Genotoxicity analysis showed that the extract produced no marked genotoxic effects at concentrations of 2.5 or 5 µg/ml in either cell type; however, at concentrations of 10 µg/ml or higher significant DNA damage was detected. The micronucleus test also demonstrated that concentrations of 10 µg/ml or higher produced clastogenic/aneugenic responses. In the Ames test, the extract induced mutagenic effects in TA98 strain of S. typhimurium with metabolic activation at all tested concentrations (2.5 to 500 µg/ml). Data indicate that, under certain experimental conditions, the fruit extract of C. oxyacantha exerts genotoxic and clastogenic/aneugenic effects in cultured human cells, and with metabolism mutagenicity occurs in bacteria cells.

Detoxification of sewage sludge by natural attenuation and implications for its use as a fertilizer on agricultural soils.

Sewage Sludges (SS) from wastewater treatment systems constitute a potential alternative to agricultural fertilizers. However, their use is limited by the presence of toxic substances that may represent significant hazards for the environment and for human health. To test the potential of natural processes to attenuate their putative toxic activities, actual SS samples from domestic sewage were buried in holes in a pollution-free environment for different periods of time, up to one year. Aqueous and organic extracts were obtained after each period of natural attenuation, and their respective toxicity was tested for estrogenic and dioxin-like activity by yeast-based bioassays (ER-RYA and AhR-RYA, respectively) and for general toxicity and teratogenicity in zebrafish embryos. Dioxin-like activity was also tested in zebrafish embryos by monitoring the induction of the marker gene cyp1a. Whereas the results showed essentially no estrogenic activity, both dioxin-like activity and embryotoxicity were observed in the initial samples, decreasing significantly after six months of attenuation. Chemical analysis of toxic SS samples showed the presence of low levels of dioxins and furans, and relatively high levels of m- and p-cresol, at concentrations that only partially justify the observed biological effects. Our data indicates the presence of largely uncharacterized hydrophilic compounds with high biological activity in SS, constituting a potential risk of groundwater pollution upon their disposal into the environment. It also shows that this potential impact may be significantly mitigated by attenuation protocols, as the one presented here.

Attesting the efficiency of monitored natural attenuation in the detoxification of sewage sludge by means of genotoxic and mutagenic bioassays.

A viable alternative to the use of sewage sludge (SS) would be using it as a reconditioner of agricultural soils, due to its high content of organic matter and nutrients. However, this solution may contaminate the soil, since SS may contain toxic substances. Monitored natural attenuation is a process that can be used in the decontamination of SS before its disposal into the environment. The effectiveness of the natural attenuation of a domestic SS was evaluated over 12 months by assays of Salmonella/microsome and micronucleus (MN) in human hepatoma cells (HepG2). Mutagenic activity was observed for the Salmonella strain TA 100, with S9, for the extracts from periods 0-6 months of natural attenuation. Genotoxic effects were observed in HepG2 cells, for 0 and 2 months, in almost all tested concentrations. Comparing obtained data by MN test to chemical analyses, it is possible to observe a coincidence between the induction of MN and the quantity of the m- and p-cresol, since these compounds were present in the initial SS and after 2 months of natural attenuation, decreasing their concentrations in samples from 6 to 12 months. The positive results obtained with Salmonella/microsome (from 6 months) suggest a combined action of other substances in SS. These results indicated that this SS, in the earlier periods tested, is potentially genotoxic and mutagenic and that its disposal can lead to severe environmental problems. Thus, the use of the studied SS as reconditioner requires pre-processing for over than 6 months of natural attenuation.

Evaluation of the genotoxicity/mutagenicity and antigenotoxicity/antimutagenicity induced by propolis and Baccharis dracunculifolia, by in vitro study with HTC cells.

The ethanolic extract of propolis, especially the Brazilian green type, is widely and mainly used for therapeutic purposes despite the lack of knowledge about its effects and its cellular mode of action. This type of propolis, derived from Baccharis dracunculifolia (alecrim-do-campo), has been extensively commercialized and the consumers use it to enhance health. This work aimed to assess the genotoxic/mutagenic and antigenotoxic/antimutagenic potentials of the ethanolic extracts of Brazilian green propolis and of B. dracunculifolia, on mammalian cells. It was not observed genotoxic and mutagenic effects by both extracts. After evaluate the exposure of the cells to each extract with a recognized mutagen, simultaneously, the results showed a significant reduction on DNA damage. The experiment carried out with a pre-incubation period was more effective than without incubation test, showing that the tested extracts were able to inactivate the mutagen before it could react with the DNA.

Induction of mitotic and chromosomal abnormalities on Allium cepa cells by pesticides imidacloprid and sulfentrazone and the mixture of them.

To evaluate the cytotoxic and genotoxic effects of low concentrations of pesticides in non-target organisms, seeds of Allium cepa were exposed for 24 h to the imidacloprid insecticide, sulfentrazone herbicide and to the mixture of them, followed by recovery periods of 48 and 72 h. Imidacloprid results indicated an indirect genotoxic effect by inducing different types of chromosome aberration (CA), mainly bridges and chromosomal adherences. Cells with micronucleus (MN) were not significant in the analyzed meristems. Moreover, the 72-h recovery tests indicated that the two lower concentrations of the insecticide (0.036 and 0.36 g L(-1)) had their genotoxic effects minimized after discontinuation of treatment, differently to the observed for the field concentration (3.6 g L(-1)). Sulfentrazone herbicide at field concentration (6 g L(-1)) caused cytotoxic effects by inducing nuclear fragmentation and inhibition of cell division. The other concentrations (0.06, 0.6 and 1.2 g L(-1)) indicated genotoxic effects for this herbicide. The concentration of 0.06 g L(-1) induced persistent effects that could be visualized both by the induction of CA in the recovery times as by the presence of MN in meristematic and F1 cells. The induction of MN by this lowest concentration was associated with the great amount of breakage, losses and chromosomal bridges. The mixture of pesticides induced genotoxic and cytotoxic effects, by reducing the MI of the cells. The chromosomal damage induced by the mixture of pesticides was not persistent to the cells, since such damage was minimized 72 h after the interruption of the exposure.

Analysis of the genotoxic potential of low concentrations of Malathion on the Allium cepa cells and rat hepatoma tissue culture.

Based on the concentration of Malathion used in the field, we evaluated the genotoxic potential of low concentrations of this insecticide on meristematic and F1 cells of Allium cepa and on rat hepatoma tissue culture (HTC cells). In the A. cepa, chromosomal aberrations (CAs), micronuclei (MN), and mitotic index (MI) were evaluated by exposing the cells at 1.5, 0.75, 0.37, and 0.18mg/mL of Malathion for 24 and 48hr of exposure and 48hr of recovery time. The results showed that all concentrations were genotoxic to A. cepa cells. However, the analysis of the MI has showed non-relevant effects. Chromosomal bridges were the CA more frequently induced, indicating the clastogenic action of Malathion. After the recovery period, the higher concentrations continued to induce genotoxic effects, unlike the observed for the lowest concentrations tested. In HTC cells, the genotoxicity of Malathion was evaluated by the MN test and the comet assay by exposing the cells at 0.09, 0.009, and 0.0009mg/5mL culture medium, for 24hr of exposure. In the comet assay, all the concentrations induced genotoxicity in the HTC cells. In the MN test, no significant induction of MN was observed. The genotoxicity induced by the low concentrations of Malathion presented in this work highlights the importance of studying the effects of low concentrations of this pesticide and demonstrates the efficiency of these two test systems for the detection of genetic damage promoted by Malathion.

Cytotoxic and genotoxic effects of two hair dyes used in the formulation of black color.

According to the International Agency for Research on Cancer (IARC), some hair dyes are considered mutagenic and carcinogenic in in vitro assays and exposed human populations. Epidemiological studies indicate that hairdressers occupationally exposed to hair dyes have a higher risk of developing bladder cancer. In Brazil, 26% of the adults use hair dye. In this study, we investigated the toxic effects of two hair dyes, Basic Red 51 (BR51) and Basic Brown 17 (BB17), which are temporary dyes of the azo group (R-N=N-R'), used in the composition of the black hair dye. To this end, MTT and trypan blue assays (cytotoxicity), comet and micronucleus assay (genotoxicity) were applied, with HepG2 cells. For cytotoxic assessment, dyes were tested in serial dilutions, being the highest concentrations those used in the commercial formula for hair dyes. For genotoxic assessment concentrations were selected according to cell viability. Results showed that both dyes induced significant cytotoxic and genotoxic effects in the cells, in concentrations much lower than those used in the commercial formula. Genotoxic effects could be related to the azo structure present in the composition of the dyes, which is known as mutagenic and carcinogenic. These results point to the hazard of the hair dye exposure to human health.

Toxicogenetic effects of low concentrations of the pesticides imidacloprid and sulfentrazone individually and in combination in in vitro tests with HepG2 cells and Salmonella typhimurium.

The insecticide imidacloprid and the herbicide sulfentrazone are two different classes of pesticides that are used for pest control in sugarcane agriculture. To evaluate the genotoxic potential of low concentrations of these two pesticides alone and in mixture, the comet assay and the micronucleus (MN) test employing fluorescence in situ hybridization (FISH) with a centromeric probe were applied in human hepatoma cell lines (HepG2), in a 24-h assay. Mutagenicity was assessed by Salmonella/microsome assay with TA98 and TA100 strains in the absence and presence of an exogenous metabolizing system (S9). The results showed significant inductions of MN in HepG2 cells by both pesticides, for all the tested concentrations. As evidenced in the comet assay, only the imidacloprid presented significant responses. When the two pesticides were associated, a significant induction of damage was observed in the HepG2 cells by the comet assay, but not by the MN test. Moreover, the MN induced by the mixtures of the pesticides appeared at lower levels than those induced by sulfentrazone and imidacloprid when tested alone. According to the FISH results, the damage induced by imidacloprid in the HepG2 cells resulted from a clastogenic action of this insecticide (76.6% of the MN did not present a centromeric signal). For the herbicide sulfentrazone and for the mixture of the pesticides, a similar frequency of MN with and without the presence of the centromeric signal (herbicide: 52.45% of the MN without centromeric signal and 47.54% of the MN with centromeric signal; mixture: 48.71% of the MN without centromeric signal and 51.42% of the MN with centromeric signal) was verified. Based on these results, it was concluded that each one of the pesticides evaluated interacts with the DNA of HepG2 cells and causes irreparable alterations in the cells. However, the combination of the pesticides showed an antagonistic effect on the cells and the damage induced was milder and not persistent in HepG2 cells. The results obtained by the Ames test did not point out significant results.

Cytotoxic and genotoxic effects of ethanolic extract of Euphorbia hyssopifolia L. on HepG2 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia hyssopifolia L. is a weed with recognized antimicrobial potential employed in Indian, Asian and Latin-American popular medicine. However, little is known with regard to its toxic potential. The present study aimed to investigate the cytotoxic and genotoxic effects of ethanolic extract of E. hyssopifolia in HepG2 cell culture. MATERIALS AND METHODS: Phytochemical screening of ethanolic extract was carried out to determine the presence of active secondary plant metabolites. Six concentrations (0.00001, 0.0001, 0.001, 0.01, 0.1 and 1.0mg/mL) of ethanolic extract were tested by the MTT assay to verify cytotoxicity. Then, genotoxic evaluations (alkaline comet assay and cytokinesis-block micronucleus assay - CBMN) were carried out in HepG2 cells with extract concentrations of 0.01, 0.1 and 1.0mg/mL. RESULTS: Mono and sesquiterpenes, triterpenes and steroids, and flavonoids were the main classes found in the phytochemical screening. Extract concentrations used in the MTT assay showed no cytotoxic activity. On the other hand, genotoxic activity was verified at 0.1 and 1.0mg/mL in the alkaline comet assay. Additionally, the 1.0mg/mL concentration induced severe cell damage leading to death in the CBMN assay, indicating a cytotoxic effect for this concentration in the latter method. CONCLUSION: The use of E. hyssopifolia extract for medicinal purposes should be avoided, because concentrations above 0.01mg/mL may pose risk to human health due to cytotoxic and/or genotoxic effects.

Assessment of cytotoxicity and AhR-mediated toxicity in tropical fresh water sediments under the influence of an oil refinery.

Oil refinery effluents contain many chemicals at variable concentrations. Therefore, it is difficult to predict potential effects on the environment. The Atibaia River (SP, Brazil), which serves as a source of water supply for many municipalities, receives the effluents of one of the biggest oil refinery of this country. The aim of this study was to identify the (eco)toxicity of fresh water sediments under the influence of this oil refinery through neutral red (cytotoxicity) and ethoxyresorufin-O-deethylase (EROD) assays (AhR-mediated toxicity) in RTL-W1 cells (derived from fish liver). Once the refinery captures the waters of Jaguarí River for the development of its activities and discharges its effluents after treatment into the Atibaia River, which then flows into Piracicaba River, sediments from both river systems were also investigated. The samples showed a high cytotoxic potential, even when compared to well-known pollution sites. However, the cytotoxicity of samples collected downstream the effluent was not higher than that of sediments collected upstream, which suggested that the refinery discharges are not the main source of pollution in those areas. No EROD activity could be recorded, which could be confirmed by chemical analyses of polycyclic aromatic hydrocarbons (PAHs) that revealed a high concentration of phenanthrene, anthracene, fluoranthene, and pyrene, which are not EROD inducers in RTL-W1 cells. In contrast, high concentrations of PAHs were found upstream the refinery effluent, corroborating cytotoxicity results from the neutral red assay. A decrease of PAHs was recorded from upstream to downstream the refinery effluent, probably due to dilution of compounds following water discharges. On the other hand, these discharges apparently contribute specifically to the amount of anthracene in the river, since an increase of anthracene concentrations could be recorded downstream the effluent. Since the extrapolation of results from acute toxicity to specific toxic effects with different modes of action is a complex task, complementary bioassays covering additional specific effects should be applied in future studies for better understanding of the overall ecotoxicity of those environments.

Genotoxicity evaluation of environmental pollutants using analysis of nucleolar alterations.

Nucleolar alterations resulting from the action of either chemical or physical agents can serve as important genotoxicity biomarkers. In this study, the efficiency of AgNOR banding technique to identify the presence of nucleoli in micronucleus and assess nucleolar alterations in aberrant cells of Allium cepa was evaluated. Seeds of this plant were exposed to both water samples from a river that receives untreated urban effluent and to the trifluralin herbicide (0.84 mg/L concentration), both analyzed in two different seasons (summer and winter seasons). Samples induced significant frequencies of chromosomal and nuclear aberrations and micronuclei, as observed in cells submitted to conventional chromosomal staining. The herbicide caused a significant increase in the number of nucleoli and micronuclei, interpreted as due to the elimination of excessive nucleolar material resulting from polyploidization. The use of the AgNOR technique enabled the identification of both the presence of the nucleolus in some micronuclei and the nucleolar organizer region (NOR) behavior of aberrant cells. The NOR-banding technique showed to be an efficient tool for studying the genotoxic effects caused by a xenobiotics and a complex environmental sample.

Evaluation of the genotoxicity of waters impacted by domestic and industrial effluents of a highly industrialized region of São Paulo State, Brazil, by the comet assay in HTC cells.

The problems that most affect the quality of the waters of rivers and lakes are associated with the discharges performed in these environments, mainly industrial and domestic effluents inappropriately treated or untreated. The comet assay is a sensitive tool and is recommended for studies of environmental biomonitoring, which aim to determine the genotoxicity potential of water pollutants. This study aimed to assess the genotoxic potential of the Ribeirão Tatu waters, region of Limeira, São Paulo (SP), by the comet assay with mammalian cells (hepatoma tissue culture (HTC)). Water samples were collected along the Ribeirão Tatu at three distinct periods: November 2008, February 2009 and August 2009, and five collection sites were established: P1, source of the stream; P2, site located downstream the urban perimeter of the municipality of Cordeirópolis and after receiving the pollution load of this city; P3, collection site located upstream the urban perimeter of the city of Limeira; P4, urban area of Limeira; and P5, rural area of Limeira, downstream the discharges of the city sewage. The results showed that for the November 2008 collection, there was no water sample-induced genotoxicity; for the February 2009 collection, the sites P1 and P2 were statistically significant in relation to the negative control (NC), and for the August 2009 collection, the site P5 was statistically significant. These results could be explained by the content of different metals during the different seasons that are under the influence of domestic, industrial and agricultural effluents and also due to the seasonality, since the water samples collected in the period of heavy rain (February 2009) presented a higher genotoxicity possibly due to the entrainment of contaminants into the bed of the stream promoted by the outflow of rainwaters. The comet assay showed to be a useful and sensitive tool in the evaluation of hydric resources impacted by pollutants of diverse origins, and a constant monitoring should be done in order to verify the influence of different factors (season, amount of contaminants) in the water quality.

Principales mecnnismos de reparación de daños en la molécula de adn / Main repair echaanisms for damages in the dna molecule

Las células cuentan con mecanismos complejos que vigilan la integridad del ADN, activando mecanismos de reparación cuando hay deficiencias o errores durante la replicación. Una consecuencia potencial de los daños son las alteraciones permanentes en la estructura del ADN que pueden generar mutaciones, transformación carcinogénica y muerte celular. Estos son atribuidos a diferentes agentes endógenos como los radicales libres de oxígeno (RLO) provenientes de la respiración, los cuales son considerados el centro de la carcinogénesis y el envejecimiento por daño genómico; agentes exógenos como la luz ultravioleta que inducen dímeros de pirimidina y la radiación ionizante que produce una gran variedad de daños sobre las bases, muchos de ellos por efecto indirecto. También se encuentran las genotoxinas presentes en los alimentos, humo de tabaco y agentes quimioterapéuticos, con grandes cualidades para alterar la estructura de la molécula ADN e interferir con su expresión. De esta manera, cerca de 10(5) lesiones espontáneas por día son inducidas en nuestros genes, en donde los mecanismos de reparación detectan daños y perturbaciones durante el crecimiento y división celular. Esto es posible gracias a las funciones específicas de reconocimiento, corrección o eliminación de daños que asegura la integridad del genoma. En este artículo se presentan los principales mecanismos de reparación del ADN, su relación y activación de acuerdo al tipo de daño.

Cells have complex mechanisms that monitor DNA integrity that activate repair mechanisms when there are deficiencies or errors during replication. A potential result of the damage is a permanent alteration in DNA structure that can generate mutations, carcinogenic transformation and cell death. These are attributed to different endogenous agents such as oxygen free radicals (OFR) from respiration, which are considered the center of carcinogenesis and aging process due to genomic damage; exogenous agents, such as ultraviolet light, induce pyrimidine dimers and ionizing radiation that produce a variety of damage on the bases, many by indirect effect. Genotoxins present in food, tobacco and chemotherapeutic agents are also found with high potential in altering the DNA molecule structure and interfering with its expression. Thus, around 10(5) spontaneous lesions are induced per day in our genes, where the repair mechanisms can detect damages and disturbances during cell growth and division. This is possible thanks to the specific recognition, correction or elimination of damage functions, ensuring the integrity of the genome. In this article the main mechanisms of DNA repair, as well as their relationship and activation according to the type of damage, are presented.

Anti-genotoxicity and anti-mutagenicity of Apis mellifera venom.

The search for substances able to inhibit and/or diminish the effects of genotoxic and mutagenic substances has been the target of several investigations performed in recent times. Hymenoptera venoms constitute a considerable source of substances with pharmacological potential. The present study aimed to evaluate the cytotoxic, genotoxic and anti-genotoxic, mutagenic and anti-mutagenic potentials of Apis mellifera venom in HepG2 cells. In this evaluation, the MTT test was applied to determine the most appropriate concentrations for the genotoxicity and mutagenicity tests. It was verified that the concentrations of 0.1, 0.05 and 0.01µg/mL were not cytotoxic, hence these concentrations were used in the experiments. For the evaluation of the genotoxic and mutagenic potential of the bee venom the comet assay and the micronucleus test were applied, respectively. The concentrations mentioned above presented both genotoxic and mutagenic potential for HepG2 cells and it was necessary to test lower concentrations of the venom (10pg/mL, 1pg/mL and 0.1pg/mL) for the anti-genotoxicity and anti-mutagenicity tests, which were performed subjecting the cells to the action of MMS (methyl methanesulfonate) in order to verify the ability of the venom to inhibit or diminish the action of this compound, which has a recognized action on the genetic material. Pre-, post-treatment and simultaneous treatment with and without incubation with the venom were performed. It was observed that the lowest three concentrations tested did not present any anti-genotoxic and anti-mutagenic activity on the cells. The use of bee venom for pharmacological purposes in treatments such as cancer must be done with extreme caution, since it was observed that even at very low concentrations the venom can induce genotoxicity and mutagenicity in human cells, as was verified for the HepG2 cells.

Application of micronucleus test and comet assay to evaluate BTEX biodegradation.

The BTEX (benzene, toluene, ethylbenzene and xylene) mixture is an environmental pollutant that has a high potential to contaminate water resources, especially groundwater. The bioremediation process by microorganisms has often been used as a tool for removing BTEX from contaminated sites. The application of biological assays is useful in evaluating the efficiency of bioremediation processes, besides identifying the toxicity of the original contaminants. It also allows identifying the effects of possible metabolites formed during the biodegradation process on test organisms. In this study, we evaluated the genotoxic and mutagenic potential of five different BTEX concentrations in rat hepatoma tissue culture (HTC) cells, using comet and micronucleus assays, before and after biodegradation. A mutagenic effect was observed for the highest concentration tested and for its respective non-biodegraded concentration. Genotoxicity was significant for all non-biodegraded concentrations and not significant for the biodegraded ones. According to our results, we can state that BTEX is mutagenic at concentrations close to its water solubility, and genotoxic even at lower concentrations, differing from some described results reported for the mixture components, when tested individually. Our results suggest a synergistic effect for the mixture and that the biodegradation process is a safe and efficient methodology to be applied at BTEX-contaminated sites.

An overview of biodiesel soil pollution: data based on cytotoxicity and genotoxicity assessments.

Biodiesel production has received considerable attention in the recent past as a nonpolluting fuel. However, this assertion has been based on its biodegradability and reduction in exhaust emissions. Assessments of water and soil biodiesel pollution are still limited. Spill simulation with biodiesel and their diesel blends in soils were carried out, aiming at analyzing their cytotoxic and genotoxic potentials. While the cytotoxicity observed may be related to diesel contaminants, the genotoxic and mutagenic effects can be ascribed to biodiesel pollutants. Thus, taking into account that our data stressed harmful effects on organisms exposed to biodiesel-polluted soils, the designation of this biofuel as an environmental-friendly fuel should be carefully reviewed to assure environmental quality.

Cytotoxicity of water-soluble fraction from biodiesel and its diesel blends to human cell lines.

The designation of biodiesel as a green fuel has increased its commercialization and use, making its fate in the environment a matter of concern. Fuel spills constitute a major source of aquatic pollution and, like diesel spills, biodiesel can produce adverse effects on aquatic environments, animals and humans. The present study assessed cytotoxic effects of water systems contaminated with neat biodiesel and its diesel blends by means of different procedures on human T cell leukemia (Jurkat) and human hepatocellular carcinoma (HepG2) cells [detection of changes in mitochondrial membrane potential (ΔΨ(m)) using tetramethylrhodamine ethyl ester (TMRE), apoptosis recognition by Annexin V and impedance real-time cell analyzer (xCELLigence™ system)]. The data obtained showed concordance across the different bioassays, with cytotoxic effects observed as a dose-dependent response only for waters contaminated with pure diesel (D100) and B5 blend, which is characterized by a mixture of 95% diesel and 5% biodiesel. The data can also lead us to hypothesize that diesel accounts for the harmful effects observed, and that biodiesel does not worsen the impacts caused by diesel pollution.