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.

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.

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.

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.

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.

Origin of nuclear and chromosomal alterations derived from the action of an aneugenic agent--Trifluralin herbicide.

Trifluralin is a herbicide capable of interfering in mitotic cell division due to either microtubule depolymerization or alteration in the concentration of calcium ions within the cell. The aim of this study was to investigate the effects of trifluralin in Allium cepa meristematic cells, evaluating the induction mechanisms of the chromosomal and nuclear aberrations. In this study, A. cepa root tips were submitted for 24h treatment to several concentrations of this herbicide and 48 h recovery post-treatment. The results showed that some concentrations of trifluralin can lead to a mitotic index inhibition, besides inducing chromosomal and nuclear alterations throughout the mitotic cycle. Some of the alterations found seem to be resulting from the herbicide action in different phases and in more than one consecutive cell cycle.