Analysis of the genotoxicity of municipal solid waste incinerator ash.

Combined bottom and fly ash obtained from a Chicago, IL, municipal solid waste incinerator (MSWI) was extracted with organic solvents, water or acidified water. The mean amounts of organic material isolated from each extraction procedure were 688.2, 91.8 and 167.7 micrograms/g MSWI ash. These extracts were evaluated for toxicity and mutagenicity in Salmonella typhimurium strains TA98 and TA100. We developed and calibrated a micropreincubation assay to evaluate small concentrations of the organic extracts. No direct-acting mutagens were found, however the acid-treated aqueous extracts were toxic. Materials isolated with methylene chloride methanol were mutagenic after hepatic microsomal activation (S9). The mutagenic potencies of the organic extract normalized to a per gram ash basis was the induction of 103.46 revertants in TA98 and 247.5 revertants in TA100. The aqueous extracts were neither toxic nor mutagenic. However, the acid-treated aqueous extract was mutagenic to TA100. The organic material isolated from the acidic extract had an induced mutagenic potency of 44.2 revertants/mg extract. Normalizing these data indicate a mutagenic potency of 7.4 revertants/g MSWI ash leached.     

Removal of mutagens from drinking water by granular activated carbon. Evaluation using bacterial mutagenicity tests

The performance of a full-scale granular activated carbon (GAC) treatment system in removing mutagens from drinking water obtained from the Ohio River has been evaluated using two bacterial mutagenicity tests. The Salmonella/microsome assay (Ames Test) and a fluctuation assay were both performed using Salmonella typhimurium strains TA98 and TA100. Influent and effluent waters were collected at two GAC adsorbers, one filled with virgin GAC and one with nearly exhausted GAC. The samples were submitted to reverse osmosis (RO) pre-concentration, sequential liquid-liquid extractions and XAD-2 resin adsorption. The RO aqueous concentrations of both influents gave positive mutagenic responses with both strains in the fluctuation assay but no activity in the Ames test. The extracts and adsorbates showed mutagenic responses in the Ames test with both strains, the highest values being observed with TA100 in the absence of metabolic activation. The summation of mutagenic activity on the basis of net revertants per liter indicated that exhausted GAC removed a substantial fraction (more than 85%) of the mutagenic activity whereas virtually complete removal was observed with virgin GAC. These data suggest that short-term mutagenicity tests may be useful in evaluating the performance of GAC or other adsorbents used in the treatment of drinking water.     

Determination of mutagenic activity in salmonella of residual fulvic acids after ozonation

Residual organics of ozonated soil and water fulvic acids were tested for mutagenicity in the Salmonella: mammalian-microsome assay. A highly ozonated soil fulvic acid extract induced His reversion in Salmonella typhimurium strain TA100 only and did not require metabolic activation for the weak mutagenic response observed.     

Mutagenic and clastogenic effects of organic extracts from the Athenian drinking water

Organic extracts of chlorinated drinking water collected from the water distribution system of Athens, Greece, were tested in parallel for their ability to cause mutations in the Salmonella His+ reversion system and sister chromatid exchanges (SCE) and chromosomal aberrations in CHO cells. They were found to induce His+ revertants in frequencies similar to those detected previously with drinking water extracts from other areas around the world. We observe that organic samples from surface drinking water induce SCE and chromatid type abnormalities in CHO cell cultures while samples collected from ground drinking water did not. These findings confirm that chlorinated surface drinking water contains substances which are mutagenic and clastogenic and may be carcinogenic in humans.     

The Salmonella mutagenicity of urban airborne particulate matter (PM2.5) from eight sites of the Emilia-Romagna regional monitoring network (Italy)

ARPA Emilia Romagna created, in 1997, a regional network for the continuous monitoring of the mutagenicity of PM(2.5) by short-term mutagenicity bioassays to guarantee a constant surveillance on the entire regional territory. The continuous monitoring of the PM mutagenicity provides essential information for a better understanding of the impact of air pollution on the health of the population, and allows one to better judge the efficiency of national and local efforts for urban air quality improvement (use of "green" petrol and ecodiesel, days during which traffic is prohibited, etc.). This article presents the results relating to the Network's activity between September 2000 and December 2002, on PM(2.5) fraction. The organic extracts of PM(2.5) were tested for mutagenicity using Salmonella tester strains TA98 and TA100 with and without metabolic activation (S9). The data obtained on the genotoxicity of air particulate extracts have revealed a constant presence of mutagenic substances adsorbed on particulate matter-with a prevalence of direct-acting mutagens than of promutagens-in a typical seasonal trend featuring higher levels in autumn-winter and lower in warmer periods of the year. In this work the evolution of PM(2.5) mutagenicity was compared with the particles, carbon monoxide (CO) and nitrogen dioxide (NO(2)) concentrations (monthly average); these comparisons revealed a quite good level of agreement on a local basis.     

Determination of halogenated organic compounds and mutagenicity testing of spent bleach liquors.

The content of organohalogenated compounds in spent bleach liquor from different bleaching stages in a sulphate and a sulphite plant has been determined by a combination of glass capillary gas chromatography, gas-liquid chromatography/mass spectrometry and neutron activation analysis. Several compounds which have not been reported before have been identified including halogenated derivatives of dimethyl-propylnaphthalenes and alkylated catechols. The unconcentrated effluents, non-polar and total extracts were tested for mutagenic activity with Ames' Salmonella test. Spent bleach liquors from most bleaching stages as well as the total effluents contained mutagenic compounds. Addition of liver microsomes for metabolic activation reduced the mutagenic activity in all stages except for the first chlorination stage in the sulphate plant. Two isomers of chloro-, bromo-, and dichloro-p-cymene previously determined in effluents from bleaching plants were synthesized from the parent molecule. Both bromo- and dichloro-p-cymene exhibited weak mutagenic activity in the Salmonella test system. Liver microsomes reduced the effect slightly. The chlorinated cymenes were found to account for up to 18% of the total organically-bound chlorine in the non-polar extracts.     

Mutagenic activity in humic water and alum flocculated humic water treated with alternative disinfectants

Mutagenic activity in Salmonella typhimurium strains TA 100 , TA 98 and TA 97 has been determined for humic water and alum flocculated humic water, treated with the alternative disinfectants chlorine, ozone, chlorine dioxide, ozone/chlorine and chlorine/chlorine dioxide.The most pronounced activity was found for chlorine treated water tested on strain TA 100 without metabolic activation (S9 mix). Ozone treatment prior to chlorination did not alter the activity, while treatment with chlorine in combination with chlorine dioxide reduced the activity to a level somewhat over the background. No mutagenic response was detected in waters treated with ozone or chlorine dioxide alone. In presence of S9 mix all water extracts studied were non-mutagenic.     

Mutagenic activity in organic concentrate from nishitakase river water in Kyoto City,and its fractions separated by using liquid-liquid fractionation and thin layer chromatography

XAD adsorption followed by elution with ethyl ether was used for recovering mutagenic substances from Nishitakase River water in Kyoto City. XAD extract recovered was separated by acid-base-neutral fractionation and tested for mutagenicity by using TA 1538 as a tester strain. The XAD extract showed strong mutagenic activity in the presence of S-9 mix, indicating the presence of frameshift type promutagens in the river water. Of the fractions separated, the neutral and basic fraction exhibited much more pronounced mutagenic activity and their contribution accounted for 98% of the mutagenic activity of the XAD extract. In order to separate mutagenic components, these active fractions were submitted to thin layer chromatography (TLC). In total 14 TLC subfractions, which had the ability to induce more than 100 net revertants/100μg of the neutral fraction or 10μg of the basic fraction, were isolated after 2 consecutive thin layer chromatographies. In this paper, the importance of varying S-9 concentration in the S-9 mix is also discussed to determine the presence or absence of synergical/antagonistic effect of chemical constituents of complex mixtures like the XAD extract.     

Mutagenic potential of laboratory chlorinated river water

Sample water was taken from a river and chlorinated at a concentration of 5 mg 1(-1) chlorine for 20 h in the laboratory in order to examine the effect of chlorination on the mutagenic potential of organic extracts that were recovered using XAD-2 resin and diethyl ether. It was found that the XAD extracts recovered from the river water showed per se strong mutagenic activity, in the presence of S-9 mix, towards Salmonella typhimurium TA 1538. When the mutagenic activity of the XAD extracts was related to the initial water volume, chlorination increased both the direct-acting and the S-9 dependent mutagenic activity. This suggests that chlorination produced de novo direct-acting and S-9 dependent mutagens and/or increased pre-existing mutagenic activities. In addition, in this study XAD extracts recovered from chlorinated and unchlorinated river water were divided into a neutral, basic and acidic fraction respectively. When the mutagenic activity of each fraction was related to the initial water volume, the neutral fraction showed the most marked increase in mutagenic activity with, and without, the S-9 mix in the chlorinated river water.     

Mutagenic assessment of Prestige fuel oil spilled on the shore and submitted to field trials of bioremediation

The mutagenicy of the slightly weathered fuel oil from the Prestige oil spill and the effects of different bioremediation products (nutrients and/or microorganisms and biodiesel) on the potential mutagenic activity of this heavy fuel oil spilled on the shore were evaluated for a period of 1 year using the Ames Salmonella assay with strains TA98, TA100, TA1535 and TA1537 in the absence and presence of exogenous metabolic activation (S9 fraction from rat liver). The in situ bioremediation experiment was performed using tiles located in the supra-littoral and intertidal zones of a beach seriously affected by the fuel oil spill. The results obtained showed the mutagenic activity of the slightly weathered fuel oil extracts at the beginning of the experiment in strain TA98 that persisted for more than 150 days in both the untreated control and treated tiles independently of the zone of the beach considered. However, after 360 days neither the control nor the treated tiles in the intertidal zone showed mutagenic activity and a weak positive response in strain TA98 was detected for the control fuel oil extracts from supra-littoral tiles. The application of biodiesel to accelerate the biodegradation of this type of fuel oil may constitute a further genotoxic hazard to the environment, since the mutagenic response achieved from the biodiesel-fuel oil mixture in the first samplings (days 0 and 30) was more potent than that obtained from the control tiles. The mutagenic activity was detected along the study with S. typhimurium TA98 in both the presence and absence of S9 microsomal fraction, but the addition of S9 fraction in the assay always increased the number of revertants induced. In general, these findings suggest that the bioremediation strategies used were not effective in eliminating the genotoxic hazard associated with this heavy fuel oil attached to rocky substrate since they did not achieve a decrease in the mutagenic response with respect to the untreated control tiles. These data also confirm that genotoxicity assays should be used to evaluate the effectiveness of bioremediation efforts associated with oil spills for a better risk assessment.     

Carcinogenic and genotoxic activity of extracts from contaminated sediments in western Lake Ontario

In order to investigate the putative association between chemical contamination in western Lake Ontario and high prevalences of fish tumors, sediments from Hamilton Harbour and Oakville Creek in Lake Ontario and reference sites in non-polluted areas of Ontario, Canada were collected and extracted for organic contaminants. Sediment extracts from Hamilton Harbour had the highest concentrations of polychlorinated biphenyls and organochlorine insecticides (ppb) and contained very high concentrations of polynuclear aromatic hydrocarbons (ppm); although the levels of these compounds varied widely with sampling location in the harbor. A sediment extract from Hamilton Harbour was mutagenic in the Ames bacterial assay, both with and without microsomal activation. High levels of aromatic DNA adducts were induced in cultured mouse C3H1OT1/2 cells after in vitro exposure to Hamilton Harbour sediment extract. In two separate carcinogenicity experiments involving a sac fry microinjection assay with rainbow trout (Oncorhynchus mykiss), Hamilton Harbour sediment extract induced hepatocellular carcinomas in fish. No hepatic neoplasms were observed in fish that had been treated with sediment extract from Oakville Creek, or with extract from a reference sediment. The significance of these results is discussed in relation to the distribution of neoplasms in feral fish within western Lake Ontario.     

A comparison of carbon black with soot

Carbon black is frequently confused with soot. In order to clarify the distinction between these materials, we have examined 6 carbon blacks and 18 samples of soot. Carbon black is composed of turbostratic colloidal aggregates which we call aciniform carbon (AC). Chimney soots from domestic wood or coal fires contain very little AC, while in diesel soots the solid particulates are essentially all AC. All soots examined contain much more extractable organic material and/or ash than does carbon black. Ames Salmonella assays were carried out on solvent extracts. The results were calculated as net revertants per mg of the original soot or carbon black. On this basis, the majority of the soot extracts assayed were at a much higher level of mutagenicity than any of the carbon black extracts. For several reasons, even greater differences in mutagen availability would be expected if the unextracted materials were compared in vivo.     

A comparison between different high volume sampling systems for collecting ambient airborne particles for mutagenicity testing and for analysis of organic compounds

Samples of urban air were collected simultaneously using different sampling systems, including electrostatic precipitation (ESP) and high volume filtration (HVF) on various filters for particle sampling and absorption on activated carbon and organic polymers for sampling of volatiles. Acetone extracts of the samples were analyzed for polycyclic aromatic hydrocarbons (PAH) and tested for mutagenicity with the Ames Salmonella/microsome assay. The results show that the concentrations of PAH found in the various particle-samples were in good agreement, whereas the mutagenic activity of these samples showed large variations. The highest mutagenic activity was found in the samples collected by ESP and on the teflon-coated glassfibre filters, whereas samples collected by high volume filtration with size-fractionation showed the lowest mutagenic activity. We do not know whether the higher activity in samples from the teflon-coated filters compared to those from ordinary glassfibre filters represent filter artifacts or if it represents a more pronounced degradation of mutagenic compounds on the non-coated glassfibre filters. Extracts from filter blanks seemed to interfere with the expression of the mutagenic activity of the positive controls, benzo[a]pyrene and nitropyrene. When sampling volatile compounds, two organic polymers, polyurethane (PUR) and XAD-2, were found suitable for collecting PAH, whereas no PAH could be detected in extracts from the activated carbon. The XAD-2 adsorbent was the most effective for sampling bicyclic PAH. None of the adsorbents yielded extracts well suited for mutagenicity testing, since blank extracts were toxic to the test bacteria. Some extracts of the PUR blanks were weakly mutagenic as well. More emphasis should be placed upon developing more efficient and unreactive adsorbents and on the adaptation of such adsorbents in samplers suited for routine use.     

Improved retention of imidacloprid (Confidor) in soils by adding vermicompost from spent grape marc

Batch sorption experiments of the insecticide imidacloprid by ten widely different Spanish soils were carried out. The sorption was studied for the active ingredient and its registered formulation Confidor. The temperature effect was studied at 15 degrees C and 25 degrees C. The addition of a vermicompost from spent grape marc (natural and ground), containing 344 g kg(-1) organic carbon, on the sorption of imidacloprid by two selected soils, a sandy loam and a silty clay loam, having organic carbon content of 3.6 g kg(-1) and 9.3 g kg(-1), respectively, was evaluated. Prior to the addition of this vermicompost, desorption isotherms with both selected soils, were also performed. The apparent hysteresis index (AHI) parameter was used to quantify sorption-desorption hysteresis. Sorption coefficients, K(d) and K(f), for the active ingredient and Confidor(R) in the different soils were similar. Sorption decreased with increasing temperature, this fact has special interest in greenhouse systems. A significant correlation (R(2)=0.965; P<0.01) between K(f) values and the organic carbon (OC) content was found, but some soils showed higher sorption coefficients than that expected from their OC values. The normalized sorption coefficients with the soil organic carbon content (K(oc)) were dispersed and low, implying that other characteristics of soils could contribute to the retention capacity as well. The spent grape marc vermicompost was an effective sorbent of this insecticide (K(f)=149). The sorption of imidacloprid increased significantly in soils amended with this vermicompost. The most pronounced effect was found in the sandy loam soil with low OC content, where the addition of 5% and 10% of vermicompost increased K(f) values by 8- and 15-fold, respectively. Soil desorption of imidacloprid was slower for the soil with the higher OC and clay content.     

Mutagenicity assay of emission extracts from wood stoves: comparison with other emission parameters

The emission from wood stoves of several types of air pollutants has been measured under standardized burning conditions with emphasis on the amount of organic compounds and determination of the mutagenic activity with the Salmonella/microsome assay. The study corroborates earlier findings that conventional wood stoves can be a significant source of hydrocarbon and tar compounds in the ambient air. The emission of mutagenic compounds comprise both compounds requiring mammalian activation and compounds which are active in the test without exogenous activation. The mutagenicity tests show that nitroaromatic compounds are present in wood stove emissions, although the emission of nitrogen oxides is low. A wood stove constructed using the downdraft principle emitted much less hydrocarbons and tar, less mutagenic components and slightly less carbon monoxide than conventional wood stoves.     

The non-steroidal anti-inflammatory drug diclofenac is readily biodegradable in agricultural soils

Diclofenac, 2-[2-[(2,6-dichlorophenyl)amino]phenyl]acetic acid, is an important non-steroidal anti-inflammatory drug widely used for human and animals to reduce inflammation and pain. Diclofenac could potentially reach agricultural lands through the application of municipal biosolids or wastewater, and in the absence of any environmental fate data, we evaluated its persistence in agricultural soils incubated in the laboratory. ¹⁴C-Diclofenac was rapidly mineralized without a lag when added to soils varying widely in texture (sandy loam, loam, clay loam). Over a range of temperature and moisture conditions extractable ¹⁴C-diclofenac residues decreased with half lives < 5 days. No extractable transformation products were detectable by HPLC. Diclofenac mineralization in the loam soil was abolished by heat sterilization. Addition of biosolids to sterile or non-sterile soil did not accelerate the dissipation of diclofenac. These findings indicate that diclofenac is readily biodegradable in agricultural soils.     

The importance of some environmental factors on the evaluation of cement stabilized arid zone soils

The effects of desiccation on the compressive strength of both soaked and unsoaked cement stabilized arid zone soils (sandy loam, silty loam and clay/clay loam), were investigated. The pre-drying of the stabilized clay/clay loam soils resulted in total strength loss on soaking, whereas the sandy loam soils still possess significant compressive strength which are however much lower than those obtained in the standard soaking test (i.e. without pre-drying). At higher cement contents (>10%), the calcareous silty loam soils showed slight increases in strength when pre-dried before soaking. Although the addition of 8% cement to most soils is capable of achieving the minimum compressive strength (1,720 KN/m²), and the maximum permissible strength reduction (20%) on soaking, possible desiccation of the stabilized soils under arid conditions could lead to unacceptable strength reductions when subjected to subsequent increase in moisture content. The results of this study highlight the importance of the use of various criteria, especially those simulating environmental conditions, in the evaluation of cement stabilized arid zone soils.     

Evaluation of the genotoxic and teratogenic potential of a municipal sludge and sludge-amended soil using the amphibian Xenopus laevis and the tobacco: Nicotiana tabacum L. var. xanthi Dulieu

The toxic, genotoxic and teratogenic potential of a municipal sewage sludge was assessed using the micronucleus assay on the larvae of the amphibian Xenopus laevis and with the tobacco somatic mutation test using the yellow-green xanthi Dulieu mutant a(1)(+)/a(1) a(2)(+)/a(2). The teratogenic potential was assessed by means of the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Various doses of the pasty sludge added to a crop soil were tested using the three bioassays. The test systems were performed either directly with sludge or sludge-amended soil samples (plant model) or with aqueous extracts (aquatic animal model). Using the tobacco model, we found no mutagenic impact of the soil amended with the sludge, perhaps because the clay-like nature of the soil, with its high adsorption capacity, may have prevented the contaminants from reaching the target. All leachates of amended soils produced a significant size reduction in Xenopus embryos. Depending on the soil/sludge ratio, some leachates were found to be genotoxic but were never teratogenic. This battery of in vivo test systems enabled us to estimate the global long-term effects under agricultural conditions with various genetic endpoints on ecologically relevant organisms characteristic of the aquatic and terrestrial compartments.     

Toxicity and mutagenicity assessment of a boron mine drainage water

This study was aimed to assess toxicity of boron mine drainage (BMD) water in K?rka borough of Turkey, which is one of the most important boron deposit sites in the world. Three complementary tests were done to assess the toxicity of BMD water. Root growth inhibition tests showed that EC₅₀ value of BMD water sample was approximately 3.85% of concentration. Furthermore, mutagenicity was analysed utilising Ames Salmonella Mutagenicity Test and Muta‐ChromoPlate kit system (Environmental Biodetection Products Inc., Ontario, Canada). Salmonella typhimurium TA 98 strain for all concentrations and Salmonella typhimurium TA100 strain for the highest concentrations of BMD water in the absence of metabolic activation significantly stimulated mutagenic activity and increased the numbers of revertant colonies for several concentrations compared to spontaneous revertant colonies. RAPD‐PCR result and the polymorphism percentage proved that the effect of boron contamination was seen on DNA level.     

Chemical characteristics and mutagenic activity of PM10 in Torino, a northern Italian city

Epidemiological studies of particulate matter (PM) have associated PM mass, as well as certain individual components of PM such as secondary particulate with adverse human health effects. For example genotoxic effects attributed to PM may relate to the content of organic compounds but also to the oxidative DNA damage generated by transition metals like iron. However the exact physiochemical mechanism by which PM produces adverse effects is not clear. The aims of this study were to evaluate (1) concentrations of PM10, (2) amounts of bioavailable iron associated with PM10, (3) amounts of secondary particulate expressed as SO4(=) and NO3(-) and (4) the mutagenic activities of PM10 organic extracts. Sampling was carried out in a meteochemical station located in Torino, a northern Italian city with high levels of PM10. The mean PM10 concentration in the considered period was 46.1+/-28.8 microg/m3, the iron mean concentration was 0.83 microg/m3 (+/-0.65 microg/m3) and the bioavailable Fe was 5.7% (+/-4.4%). The data showed that secondary particulate matter (as sum of sulfates and nitrates) constituted about 47% of PM10 total mass. Both iron and secondary species concentrations were positively associated with PM10 levels. Seasonal variations of PM10 concentration, iron level and secondary species amount were significant. Samples were tested for mutagenicity with Salmonella typhimurium strains TA98 and TA100, with and without metabolic activation and a positive response was observed especially for TA98. There were positive statistical associations between mutagenicity and PM10, bioavailable iron, sulfates and nitrates concentrations. Therefore, these results showed the usefulness of this biological approach for monitoring PM10.