Phyto-genotoxicity assessment of different associations between sludges from Water and Sewage Treatment Plants, before and after the bioremediation process.

Water Treatment Plants (WTP) and Sewage Treatment Plants (STP) generate residues known as sludge (WS and SS, respectively). SS and WS present some positive characteristics for reuse in agriculture. The aim of the present study was to evaluate, using the Allium cepa test, the effectiveness of the bioremediation process in the detoxification of SS and WS sludges. In this study, the phytotoxic, cytotoxic, genotoxic, and mutagenic potentials of pure sludge samples (WS and SS) were evaluated, as well as the association of these two sludges with soil (S), before and after the bioremediation process. In the T0 period (before undergoing bioremediation), the SS, SS + S, and SS + WS samples totally inhibited the germination of A. cepa, proving the high phytotoxic potential of these samples. For the T1 period (after 6 months of bioremediation), phytotoxicity was observed for the SS, SS + S, SS + WS, and SS + WS + S samples, but there was not a complete inhibition of germination and radicles growth, allowing the evaluation of the other parameters (cytogenotoxic and mutagenic potential). No cytotoxicity was observed for any sample, both in T0 and T1. As for the genotoxicity parameter, a significant result was observed for the pure WS sample in T0 and for all samples in T1, when compared to NC. The genotoxic alteration most found in meristematic cells exposed to treatments was of binucleated cells. Mutagenic potential was also observed for samples of WS and WS + S in T0. From this study, we can conclude that, after six months of bioremediation, despite the SS phytotoxicity being reduced, all samples were genotoxic to the A. cepa organism test.

Toxicity of the sawdust used for phosphorus recovery in a eutrophic reservoir: experiments with Lactuca sativa and Allium cepa.

Eutrophication is one of the environmental problems arising from the increase of essential nutrient concentrations, mainly phosphorus and nitrogen. In contrast to excess phosphorus, the depletion of phosphate rock deposits used for the production of fertilizers compromises the food supply. Therefore, the development of technologies that propose the recovery of the phosphorus contained in eutrophic environments for its later use for agricultural fertilization purposes is very important to ensure global food security. This work aimed to evaluate the toxic potential of the sawdust (biosorbent previously used for phosphorus adsorption) in order to enable its application in agriculture. For this, toxicity experiments with Lactuca sativa (lettuce) and Allium cepa (onion) seeds were performed. The phytotoxic potential was assessed by means of the seed germination index and physiological parameters such as radicle and hypocotyl growth. Cytotoxicity, genotoxicity, and mutagenicity tests were also performed on onion seeds. From statistical tests, it was possible to affirm that the sawdust did not promote inhibition of seed germination and radicle and hypocotyl growth. No genotoxicity, cytotoxicity and, mutagenicity were observed, which allowed to state that the sawdust is not toxic to the onion species, which reinforces the possibility of application of the biosorbent for soil fertilization purposes. Therefore, the use of sawdust for phosphorus biosorption with the subsequent agricultural application is promising and quite important from a global food security point of view.

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.

DNA damages induced by both endotoxin and exotoxin produced by cyanobacteria.

Cyanobacteria are prokaryotes involved in the contamination of aquatic environments since they release toxins that are highly potent and dangerous for living organisms. Prokaryotes produce endo and exotoxins, among others. Exotoxins are highly toxic, while endotoxins have milder toxic effects. The present study evaluated the cytotoxicogenetic potency of both toxins studying them in different concentrations of cyanobacterial biomasses (1 µg/L, 1.5 µg/L, 2 µg/L), to assess the amount of exotoxin present in the cultured medium in which the cyanobacteria were grown. For this evaluation, we used an extract taken from the medium in a concentration of 10%. Our results showed that genotoxic and mutagenic changes in Allium cepa could be observed in all of the varying concentrations of biomass (endotoxin action) and also in the medium induced with exotoxin. Even at low concentrations, these toxins were highly effective at triggering changes in the DNA molecules of organisms exposed to them. This information is highly significant when considering environmental contamination caused by cyanobacteria blooms, since the results of this study show that these toxins may not only kill organisms when found in high concentrations, but also induce mutations when found in low concentrations. Since these mutations are expressed later on in the organisms, it is impossible to associate the observed effect with the event that induced the damage.

Comparison of the toxicogenetic potential of sewage sludges from different treatment processes focusing agricultural use.

A problem that has been dragging in recent decades is the final disposal of the waste produced in the wastewater treatment process. In addition to its high amount of organic matter and nutrients, this waste, known as sewage sludge (SS), may also contain toxic compounds that, when in the environment, can cause deleterious effects to organisms and lead to severe and irreversible consequences to human health. In order to understand the potential of inducing cellular and chromosomal instabilities, the species Allium cepa was employed to assess the presence of toxic agents in SS samples. Seeds of A. cepa were exposed to several dilutions of aqueous extract of SSs from 5 wastewater treatment plants (WWTPs), whose characteristics of treated sewage and the technologies employed differ among them. The results obtained showed that all the studied SSs induced significant genotoxic and mutagenic alterations, even in smaller dilutions tested. With these results, it was also possible to observe that SSs from WWTPs that present system of activated sludge and receive sewage of industrial origin induced a greater number of toxicogenetic alterations in the test organism. The high frequencies of chromosomal and nuclear aberrations observed, induced by contaminants present in the SS, represent worrying results because it proves a direct action of this agent on the genetic material of the exposed organism. Therefore, the agronomic application of SS in agriculture requires additional and more effective technologies in order to promote its complete decontamination and its safe disposal in the environment.

Genetic toxicity of water contaminated by microcystins collected during a cyanobacteria bloom.

Microcystin-LR (MCLR) is a toxin mainly produced by Microcystis aeruginosa, cyanobacteria most commonly found in eutrophic environments. Cyanobacteria blooms have affected Salto Grande reservoir (Americana, State of São Paulo/Brazil) for several decades, often observed during periods of drought. In this study, the genotoxic effects of MCLR (95% purity) and water samples contaminated by this toxin were evaluated during cyanobacteria bloom using assays with the test organism Allium cepa. The results showed genotoxic action for pure microcystin and cytotoxic, genotoxic and mutagenic action for water samples collected during flowering. Chromosomal aberration assays have shown that MCLR induces chromosomal breaks that persist in the daughter cells as MN. Therefore, it is possible to infer a clastogenic action for this toxin. The MCLR present in the environmental samples was shown to be more cytogenotoxic for the cells than the different concentrations tested in this study with the pure substance. This amplified toxic action can be related to a synergistic effect between the MCLR and other compounds present in the environmental samples. The genotoxicity studies with MCLR show inconsistent and inconclusive results, so this toxin needs to be better investigated in order to obtain further information about the action mode of it is on the biological system.

Ecotoxicological and microbiological assessment of sewage sludge associated with sugarcane bagasse.

Sewage sludge (SS) obtained after sewage treatment process may contain several toxic substances. Bioremediation can decrease the toxicity of the sludge, mainly when it is associated with stimulant agents, such as sugarcane bagasse (B). Samples of pure SS (SSP); SS+B; SS+Soil; and SS+B+Soil were bioremediated for 1, 3, and 6 months (T1, T2, and T3, respectively). After each period, the cytotoxic, genotoxic, and mutagenic potentials of the solid samples and their respective aqueous extracts (aqueous eluate and percolate water) were evaluated by the Allium cepa test. A microbiological analysis of the samples was also performed after each period tested. All solid samples of SS+B (in T1, T2, and T3) and the solid sample of SSP (treatment T3) showed a significant decrease of cell division (cytotoxic effects). The aqueous eluate extracts of SS+B (T1 and T3) and SSP (T2 and T3) induced cytotoxic effect. The solid sample of SS+B (T2 and T3) and aqueous extracts of SSP (T1) were genotoxic, indicating a harmful effect of SS on A. cepa, even after 6 months of bioremediation. There was an alternation in the microbial community both in diversity and in abundance, with the predominance of nonfermenting gram-negative bacilli. The tested bioremediation periods were not sufficient for the complete detoxification of SS, and the use of B did not seem to contribute to the degradation of the pollutants to inert compounds. These data emphasize that a specific relationship should exist between the sludge characteristic and the biostimulating agent used to promote a more efficient bioremediation. These results suggest the necessity to study longer periods of biodegradation and the use of other decomposing agents for greater safety and sustainability for the agricultural use of this residue.

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.

In vitro and in vivo investigation of the genotoxic potential of waters from rivers under the influence of a petroleum refinery (São Paulo State - Brazil).

In recent years concern about the chemical composition of wastewater generated by the oil refining industry has increased, even after its treatment. These wastewaters contain substances that can harm both the entire aquatic ecosystem and the health of any exposed organisms. The aim of this study was to evaluate the genotoxic and mutagenic potentials of the effluent generated by the largest Brazilian petroleum refinery, the effectiveness of the treatments used by the refinery, and whether its effluent can compromise the water quality of the river where it is discarded. Chromosomal aberration and micronucleus assays were performed in Allium cepa and micronucleus test in mammalian cell culture (CHO-K1). The samples were collected in three sites at the refinery: one site on the Jaguari River and two sites on the Atibaia Rivers (upstream and downstream of the discharged effluent), under three different climatic conditions. Tests with A. cepa showed increased frequencies of chromosomal aberrations and micronuclei in meristematic cells for the effluent after physico-chemical treatment, but the samples after treatment biological and stabilization pond presented none of these abnormalities. It was observed that the induced damage in the meristematic cells was not observed in the F1 cells of A. cepa roots. The micronucleus test performed with mammalian cell culture also indicated that the effluent, after physico-chemical treatment, induced a significant increase in micronucleus frequencies. Plant and hamster cells exposed to the other samples collected inside the refinery and in the Jaguari and Atibaia Rivers did not present evidence of genotoxicity and mutagenicity in the tests performed. This study showed that the effluent treated carried out by the refinery (biological treatment followed by a stabilization pond) proved to be efficient for the removal of the toxic load still present after the physico-chemical treatment, since no change in the quality of the Atibaia River was observed. However, because this is an industry with a high production of effluent with toxic potential, its effluents must be constantly monitored, so that there is no compromise of the water quality of the receiving river.

Toxicological effects of the waste of the sugarcane industry, used as agricultural fertilizer, on the test system Allium cepa.

Sugarcane is cultivated in tropical countries for sugar and ethanol production. In Brazil, this culture is among the most profitable with a production of 658.7 million tons/harvest. Sugarcane filter cake (SCFC) is a waste rich in organic matter and micronutrients, but also contains toxic metals. As it has been used as fertilizer and there is not enough knowledge about its environmental impacts, this work assessed the genotoxicogenetic effects of raw SCFC and associations with soil and sugarcane bagasse (SCB), by Allium cepa tests. Six associations of SCFC + soil and five associations of SCFC + soil + SCB were tested at three moments of degradation: initial (T0), 3 and 6 months (T1 and T2). Genotoxicogenetic assays were performed with solid substrates of these associations and with their respective aqueous extracts. Chemical analysis showed a decrease in metals, total organic carbon and nitrogen after 6 months of biodegradation, complying with Brazilian laws. In general, the combination of SCFC + soil + SCB was better than using only SCFC. T0 solubilized samples of different associations composed by highest quantities of SCFC inhibited the MI and induced CA without presenting mutagenicity (except for 75%-SCFC + soil + SCB). T1 samples showed more cytotoxicity than T0 samples, and also presented genotoxic and mutagenic effects. Solid substrate and solubilized associations of SCFC + soil + SCB of T2 samples had no toxicity. These results suggest 6 months of biodegradation and the SCB adding as effective to reduce toxicogenetic effects induced by SCFC. Also, small proportions of SCFC interfered less on the A. cepa test-system when compared with those containing high quantities of residue.

Assessment of the cytotoxic, genotoxic and mutagenic effects of the commercial black dye in Allium cepa cells before and after bacterial biodegradation treatment.

The present study evaluated the cytotoxic, genotoxic and mutagenic actions of different concentrations (50 and 200 µg/L) of BDCP (Black Dye Commercial Product) used by textile industries, before and after bacterial biodegradation, by the conventional staining cytogenetic technique and NOR-banding in Allium cepa cells. Differences in the chromosomal and nuclear aberrations and alterations in the number of nucleoli were observed in cells exposed to BDCP with and without the microbial treatment. The significant frequencies of chromosome and nuclear aberrations noted in the tests with bacterially biodegraded BDCP indicate that the metabolites generated by degradation are more genotoxic than the chemical itself. Losses of genetic material characterize a type of alteration that was mainly associated with the action of the original BDCP, whereas chromosome stickiness, nuclear buds and binucleated cells were the aberrations that were preferentially induced by BDCP metabolites after biodegradation. The significant frequencies of cell death observed in the tests with biodegraded BDCP also show the cytotoxic effects of the BDCP metabolites. The reduction in the total frequency of altered cells after the recovery treatments showed that the test organism A. cepa has the ability to recover from damage induced by BDCP and its metabolites after the exposure conditions are normalized.

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 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.

Genotoxic potential of the latex from cotton-leaf physicnut (Jatropha gossypiifolia L.).

Jatropha gossypiifolia L. (Euphorbiaceae), popularly known as cotton-leaf physicnut, is a milky shrub notable for its medicinal properties. The present study aimed to evaluate the toxic, cytotoxic and genotoxic effects of the latex of J. gossypiifolia, using Allium cepa L. as test system. Seeds of A. cepa were exposed to five concentrations of the latex (1.25; 2.5; 5; 10 and 20 mL/L) in order to evaluate parameters of toxicity (evaluation of root growth), cytotoxicity (mitotic index frequency) and genotoxicity (frequency of chromosome alterations). The latex showed a significant decrease in root mean growth value as well as mitotic index for the tested concentrations, except for 1.25 mL/L, when compared to results from the negative control. The 1.25, 2.5 and 5 mL/L concentrations induced significant chromo-some adherences, C-metaphases and/or chromosome bridges, as genotoxic effects. The significant frequency of chromosome bridges also indicated mutagenic potential for chromosomes of J. gossypiifolia as discussed in the paper. Considering that the latex is used in popular therapies, and that the test system A. cepa presents good correlation with tests carried out in mammals, it can be pointed out that its use for medicinal purposes may be harmful to human health especially if ingested.

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.

Could contaminant induced mutations lead to a genetic diversity overestimation?

Contaminant driven genetic erosion reported through the inspection of selectable traits can be underestimated using neutral markers. This divergence was previously reported in the aquatic system of an abandoned pyrite mine. The most sensitive genotypes of the microcrustacean cladoceran Daphnia longispina were found to be lacking in the impacted reservoir near the entrance of the metal rich acid mine drainage (AMD). Since that divergence could be, at least partially, accounted for by mutagenicity and genotoxicity of the AMD, the present study aimed at providing such a characterization. The Allium cepa chromosomal aberration assay, using root meristematic cells, was carried out, by exposing seeds to 100, 10, 1, and 0.1 % of the local AMD. Chromosomal aberrations, cell division phases and cell death were quantified after the AMD exposure and after 24 and 48 h recovery periods. The AMD revealed to be mutagenic and genotoxic, even after diluting it to 1 and 0.1 %. Dilutions within this range were previously found to be below the lethality threshold and to elicit sublethal effects on reproduction of locally collected D. longispina clonal lineages Significant mutagenic effects (micronuclei and chromosomal breaks) were also found at 0.1 % AMD, supporting that exposure may induce permanent genetic alterations. Recovery tests showed that AMD genotoxic effects persisted after the exposure.

Cellular damages in the Allium cepa test system, caused by BTEX mixture prior and after biodegradation process.

Petroleum and derivatives have been considered one of the main environmental contaminants. Among petroleum derivatives, the volatile organic compounds benzene, toluene, ethylbenzene and xylene (BTEX) represent a major concern due to their toxicity and easy accumulation in groundwater. Biodegradation methods seem to be suitable tools for the clean-up of BTEX contaminants from groundwater. Genotoxic and mutagenic potential of BTEX prior and after biodegradation process was evaluated through analyses of chromosomal aberrations and MN test in meristematic and F(1) root cells using the Allium cepa test system. Seeds of A. cepa were germinated into five concentrations of BTEX, non-biodegraded and biodegraded, in ultra-pure water (negative control), in MMS 4×10(-4)M (positive control) and in culture medium used in the biodegradation (blank biodegradation control). Results showed a significant frequency of both chromosomal and nuclear aberrations. The micronucleus (MN) frequency in meristematic cells was significant for most of tested samples. However, MN was not present in significant levels in the F(1) cells, suggesting that there was no permanent damage for the meristematic cell. The BTEX effects were significantly reduced in the biodegraded samples when compared to the respective non-biodegraded concentrations. Therefore, in this study, the biodegradation process showed to be a reliable and effective alternative to treat BTEX-contaminated waters. Based on our results and available data, the BTEX toxicity could also be related to a synergistic effect of its compounds.

Genotoxicity and mutagenicity of water samples from the Monjolinho River (Brazil) after receiving untreated effluents.

Cytotoxicity, genotoxicity and mutagenicity assays, using the Allium cepa test-system, were carried out in order to evaluate the effects of domestic and industrial effluents in the Monjolinho River in different seasons of the year. In the summer and intermediate seasons, chromosome aberration, micronuclei, cell death and inhibition of the mitotic index were observed in water samples collected at different sites. In the winter, either chromosome or cellular alterations were not observed. Through chemical analysis, we infer that the excessive metals such as Pb, Ni and Cu were mainly responsible for the effects observed in A. cepa cells. Limnologic analysis like electrical conductivity, dissolved oxygen and the presence of nitrogen and phosphated compounds showed that the river's contamination is due to organic matter discharge along its course. Moreover we note that this river had a higher self-depurative capacity at the end of its course, before its confluence with the Jacaré-Guaçu River.