RESUMO
Urban particulate matter (UPM) contributes to lung cancer incidence. Here, we have studied the mutagenic activity and DNA adduct-forming ability of fractionated UPM extractable organic matter (EOM). UPM was collected with a high-volume sampler in June 2004 at two sites, one at street level adjacent to a roadway and the other inside a park within the urban area of the city of São Paulo, Brazil. UPM was extracted using dichloromethane, and the resulting EOM was separated by HPLC to obtain PAH, nitro-PAH, and oxy-PAH fractions which were tested for mutagenicity with the Salmonella strains TA98 and YG1041 with and without S9 metabolic activation. The PAH fraction from both sites showed negligible mutagenic activity in both strains. The highest mutagenic activity was found for the nitro-PAH fraction using YG1041 without metabolic activation; however, results were comparable for both sites. The nitro-PAH and oxy-PAH fractions were incubated with calf thymus DNA under reductive conditions appropriate for the activation of nitro aromatic compounds, then DNA adduct patterns and levels were determined with thin-layer chromatography (TLC) 32P-postlabeling method using two enrichment procedures-nuclease P1 digestion and butanol extraction. Reductively activated fractions from both sites produced diagonal radioactive zones (DRZ) of putative aromatic DNA adducts on thin layer plates with both enrichment procedures. No such DRZ were observed in control experiments using fractions from unexposed filters or from incubations without activating system. Total adduct levels produced by the nitro-PAH fractions were similar for both sites ranging from 30 to 45 adducts per 10(8) normal nucleotides. In contrast, the DNA binding of reductively activated oxy-PAH fractions was three times higher and the adduct pattern consisted of multiple discrete spots along the diagonal line on the thin layer plates. However, DNA adduct levels were not significantly different between the sampling sites. Both samples presented the same levels of mutagenic activity. The response in the Salmonella assay was typical of nitroaromatics. Although, more mutagenic activity was related to the nitro-PAH fraction in the Salmonella assay, the oxy-PAH fractions showed the highest DNA adduct levels. More studies are needed to elucidate the nature of the genotoxicants occurring in São Paulo atmospheric samples.
Assuntos
Adutos de DNA/metabolismo , Nitrocompostos/toxicidade , Compostos de Oxigênio/toxicidade , Material Particulado/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Atmosfera/química , Brasil , Cidades , Testes de Mutagenicidade , Hidrocarbonetos Policíclicos Aromáticos/química , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genéticaRESUMO
The water produced by the Cristais River Drinking Water Treatment Plant (CR-DWTP) repeatedly produced mutagenic responses that could not be explained by the presence of disinfection byproducts (DBPs) generated by the reaction of humic acids and chlorine. In order to determine the possible role of chlorinated dye products in this mutagenic activity, solutions of a black dye commercial product (BDCP) composed of C.I. Disperse Blue 373, C.I. Disperse Orange 37, C.I. Disperse Violet 93, and chemically reduced BDCP (R-BDCP) were chlorinated in a manner similar to that used by the CR-DWTP. The resulting solutions were extracted with XAD-4 along with one drinking water sample collected from the CR-DWTP. All extracts showed mutagenic activity in the Salmonella/microsome assay. Dye components of the BDCP as well as its reduced chlorinated (CI-R-BDCP) derivative were detected in the drinking water sample by analysis with a high performance liquid chromatography/diode array detector (HPLC/DAD). The mutagenicity results of these products suggest that they are, at least in part, accounting for the mutagenic activity detected in the drinking water samples from the Cristais River. The data obtained in this study have environmental and health implications because the chlorination of the BDCP and the R-BDCP leads to the formation of mutagenic compounds (CI-BDCP and CI-R-BDCP), which are potentially important disinfection byproducts that can contaminate the drinking water as well as the environment.
Assuntos
Cloro/química , Corantes/farmacologia , Purificação da Água , Cloro/farmacologia , Cromatografia , Cromatografia Líquida de Alta Pressão , Meio Ambiente , Corantes de Alimentos , Modelos Químicos , Mutagênese , Testes de Mutagenicidade , Rios , Salmonella/metabolismo , Fatores de Tempo , Poluentes da Água , Abastecimento de ÁguaRESUMO
Mutation spectra analysis can provide important information about the types of genotoxic compounds that can be present in environmental samples. In this study, we used the TA7000 base-specific Salmonella typhimurium tester strains to characterize water samples from two drinking water treatment plants (DWTPs) in São Paulo, Brazil. Because of the small sample sizes of these environmental samples, the use of the microsuspension protocol was necessary. Acidic extracts of drinking water samples from the two DWTPs gave similar responses in the TA7000 strains and caused primarily CG to AT transversions. It is likely that halogenated disinfection by-products, generated during the chlorination of water, are causing the response seen with the TA7000 strains.
Assuntos
Cloro/toxicidade , Mutação/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacos , Abastecimento de Água/análise , Animais , Brasil , Cloro/química , Relação Dose-Resposta a Droga , Genótipo , Hidrocarbonetos Clorados/toxicidade , Microssomos Hepáticos/química , Testes de Mutagenicidade , Mutação/genética , Mutação Puntual/efeitos dos fármacos , Mutação Puntual/genética , Ratos , Ratos Sprague-Dawley , Salmonella typhimurium/genéticaRESUMO
To verify whether dyes emitted within the discharge of a dye processing plant were contributing to the mutagenicity repeatedly found in the Cristais River, Sao Paulo, Brazil, we chemically characterized the following mutagenic samples: the treated industrial effluent, raw and treated water, and the sludge produced by a Drinking Water Treatment Plant (DWTP) located approximately 6 km from the industrial discharge. Considering that 20% of the dyes used for coloring activities might be lost to wastewaters and knowing that several dyes have mutagenic activity, we decided to analyze the samples for the presence of dyes. Thin layer chromatographic analysis indicated the presence of three prevalent dyes in all samples, except for the drinking water. This combination of dyes corresponded to a commercial product used by the industry, and it tested positive in the Salmonella assay. The structures of the dye components were determined using proton magnetic resonance and mass spectrometric (MS) methods, and the dyes were tested for mutagenicity. The blue component was identified as the C.I. Disperse Blue 373, the violet as C.I. Disperse Violet 93, and the orange as C.I. Disperse Orange 37. The dyes showed mutagenic responses of 6300, 4600, and 280 revertants/microg for YG1041 with S9 respectively. A bioassay-directed fractionation/chemical analysis showed that the C.I. Disperse Blue 373 contributed 55% of the mutagenic activity of the DWTP sludge. We showed that these dyes contributed to the mutagenic activity found in the Cristais River environmental samples analyzed and are indirectly affecting the quality of the related drinking water. Therefore, we believe that this type of discharge should be more thoroughly characterized chemically and toxicologically. Additionally, human and ecological risks associated with the release of dye processing plant effluents should be more fully investigated, especially where the resultant water is taken for human consumption.
Assuntos
Compostos Azo/toxicidade , Corantes/toxicidade , Mutagênicos/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Compostos Azo/química , Bioensaio , Brasil , Cromatografia em Camada Fina , Corantes/química , Água Doce/análise , Técnicas In Vitro , Resíduos Industriais/efeitos adversos , Resíduos Industriais/análise , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Microssomos Hepáticos/metabolismo , Testes de Mutagenicidade , Mutagênicos/química , Ratos , Salmonella/efeitos dos fármacos , Indústria Têxtil , Poluentes Químicos da Água/análise , Abastecimento de Água/análiseRESUMO
Textile dyes can enter the water ecosystem through wastewater discharges potentially exposing humans through the consumption of water and food. The commercial disperse dye product CI Disperse Blue 291 containing the aminoazobenzene 2-[(2-bromo-4,6-dinitrophenyl)azo]-5-(diethylamino)-4-methoxyacetanilide (CAS registry no. 56548-64-2) was tested for mutagenic activity in the Salmonella assay. We used strains with different levels of nitroreductase and O-acetyltransferase (i.e., TA98DNP6, YG1024, and YG1041) that are relevant enzymes in the activation of nitrocompounds by the intestinal microflora. The commercial product tested also was mutagenic for TA1537, TA1538, TA98 and TA100. Presence of the pKM101 plasmid and the addition of S9 enhanced the mutagenic response. Specialized strains showed that both nitroreductase and O-acetyltransferase are important in activation of the product. The highest potency obtained was 240 revertants per microgram for YG1041 in the presence of S9. Besides being able to cause frameshift mutations (hisd3052), the dye was able to cause all types of base pair substitution with a preference for TA to AT; CG to TA and CG to AT changes. With these results clearly showing that the bacterial nitroreductase and O-acetyltransferase metabolites of this compound are mutagenic, there is a need to test this dye using in vivo systems to verify possible adverse effects of this product in mammalian tissues.
Assuntos
Acetanilidas/toxicidade , Compostos Azo/toxicidade , Corantes/toxicidade , Testes de Mutagenicidade/métodos , Poluentes Químicos da Água/toxicidade , Acetiltransferases/metabolismo , Relação Dose-Resposta a Droga , Mutação da Fase de Leitura , Água Doce , Humanos , Nitrorredutases/metabolismo , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genéticaRESUMO
In the routine São Paulo state (Brazil) surface water quality-monitoring program, which includes the Salmonella microsome mutagenicity assay as one of its parameters, a river where water is taken and treated for drinking water purposes has repeatedly shown mutagenic activity. A textile dyeing facility employing azo-type dyes was the only identifiable source of mutagenic compounds. We extracted the river and drinking water samples with XAD4 at neutral and acidic pH and with blue rayon, which selectively adsorbs polycyclic compounds. We tested the industrial effluent, raw, and treated water and sediment samples with YG1041 and YG1042 and compared the results with the TA98 and TA100 strains. The elevated mutagenicity detected with YG-strains suggested that nitroaromatics and/or aromatic amines were causing the mutagenicity detected in the samples analyzed. Positive responses for the blue rayon extracts indicated that mutagenic polycyclic compounds were present in the water samples analyzed. The mutagen or mixture of mutagens present in the effluent and water samples cause mainly frameshift mutations and are positive with and without metabolic activation. The Salmonella assay combined with different extraction procedures proved to be very useful in the identification of the origin of the pollution and in the identification of the classes of chemical compounds causing the mutagenic activity in the river analyzed.