The influence of application rate on the bacterial mutagenicity of soil amended with municipal sewage sludge

The mutagenic potential of two soils amended with a municipal sewage sludge at two application rates was monitored over a 2-year period using Salmonella/microsome mutagenicity assay. Samples were collected from undisturbed monolith lysimeters of Weswood sandy clay (Fluventic Ustochrept) and Padina sandy loam (Grossarenic Paleustalf) amended with dried sewage sludge at 50 and 100 Mg/ha. Soil samples were collected and sequentially extracted with methylene chloride and methanol. The residues from these extracts were tested for mutagenicity at five doses with and without metabolic activation in Salmonella strain TA98. In general, the mutagenic potential of the amended soils of both application rates for the first 8 weeks following sludge application increased and then slowly decreased. The maximum mutagenic response observed in the soil extracts was 222 revertants at a dose of 10 mg of residue. This response was induced by the methanol extract from the Weswood soil collected 56 days after the application of 50 Mg/ha sewage sludge as compared to the 100 Mg/ha application which induced 202 revertants/mg. The mutagenicity of all fractions extracted from the sludge-amended soil at both application rates collected 717 days after application were not appreciably different from extracts from the unamended soils. The data indicate that chemicals that were mutagenic in bacteria persist in the soil and that at the higher application rates, as much as 2 years may be required for the mutagenic potential of the soil to return to background levels.     

The aqueous sulfite-nitrite system: Investigations on the mutagenicity of some known products

Some known reaction products of the two commonly used food additives, sulfite and nitrite, were examined for mutagenicity using the Salmonella/mammalian-microsome test. Potassium nitrosodisulfonate, potassium aminetrisulfonate and potassium hydroxylaminemonosulfonate were not mutagenic over a dose range of 0.01 – 10 mg/plate in the strains his G 46, TA 100 and TA 98. Potassium hydroxylaminedisulfonate showed a weak mutagenic activity in his G 46 and TA 100 with microsomal activation. Hydroxylamine-O-sulfonic acid was only weakly mutagenic in the excision-repair proficient strain his G 46 in the presence of S9.     

长江、嘉陵江(重庆段)源水有机提取物的致突变活性及其季节变化

为了确定并比较重庆主城区段长江、嘉陵江源水有机提取物的致突变性及其季节变化规律，分别于春、夏、冬季采用GDX－120大孔树脂，对位于城区上游、城区中段、城区下游以长江、嘉陵江源水的5个水厂的进厂水进行了有机物的浓缩提取。提取物的致突变活性采用经典的Ames试验平板掺入法评估，测试菌株为TA98及TA100，同时做加与不加S9的比较。结果显示，嘉陵江及长江源水的有机提取物均有不同程度的致突变活性。嘉陵江源水明显大于长江源水，城区中段源水明显大于上游段及下游段源水。多数断面显示平水期致突变活较为显著并且移码型致突变性大于碱基置换型致密变性。研究结果提示，城市污染源已导致长江、嘉陵江源水具备致突变活性，控制两江沿岸的各种水污染源已成为当务之急。     

Residues and mutagenicity of captan applied to apple trees and potential human exposure

The fungicide captan (cis-N-((trichloromethyl)thio) 4-cyclo-hexene-1,2-dicarboximide) was applied at the rate of 2.4 g/l to apple trees (c.v. Golden Delicious) individually or as part of a standard treatment program where it was applied eight times during the growing season together with several pesticides. Leaf samples (100 discs of 2.2 cm diameter) were collected from treated and control trees before treatment and at 0, 1, 3, 7, 14, 28, 56, 90 and 112 days after treatment. Fruit samples were taken at mid-season (56 days) and at harvest (112 days). The objective of this study was to determine the captan residue and mutagenicity of leaf and fruit extracts to ascertain the potential health hazard to agricultural workers in these orchards. Surface residues were extracted from leaves and fruits with methylene chloride. These extracts were subsequently analyzed for captan by gas-liquid chromatography (GLC) utilizing an electron-capture detector, and for mutagenicity with two strains (TA98 and TA100) of Salmonella typhimurium, with and without microsomal enzyme activation. Positive mutagenic effects were observed with strain TA100 at 0-14 days post spray, even with extracts from one leaf disc's surface (3.8 cm2) of the single treatment. Captan residues in these samples indicated a decline from 9.3 micrograms/cm2 at 0 days to 0.80 micrograms/cm2 at 14 days and a trace after 112 days. With the standard treatment, in which captan was incorporated eight times in the program starting at the 7-day interval, leaf extracts showed mutagenic activity at 7, 14, 28 and 90 days. Captan residues at these intervals were 11.4, 5.0, 4.1 and 3.4 micrograms/cm2, respectively. Fruit sample extracts of the standard spray were mutagenic to the tester strains TA100 and TA98 both at mid-season and at harvest. Residues of captan on fruits declined from 10.4 micrograms/cm2 at mid-season to 1.1 micrograms/cm2 at harvest. No mutagenic activity was detected with extracts from fruit samples from the single captan application.     

The Mutagenic Activity of Particulate Organic Matter Collected with a Dilution Sampler at Coal-Fired Power Plants

The Deep Creek Lake Study of 1983 provided an opportunity to obtain emission samples from coal-fired power plants with a dilution sampler for mutagenicity testing. Stack and ambient samples of particulate matter were collected with a dilution sampler at three coal-fired power plants in West Virginia. Samples were sequentially extracted with cyclohexane (CX), dichloromethane (DCM) and acetone (ACE) and tested for mutagenicity in the Ames Salmonella/microsome assay using TA98 (-S9). For the stack samples, the CX, DCM and ACE fractions constituted 1.0, 0.7 and 98.1 percent of the total extractable organic material (EOM), respectively, compared to 28.5, 7.4 and 64.1 percent for the ambient samples. In contrast, the mutagenic activity of the organic fractions was concentrated in the CX and DCM fractions.

The cyclohexane- and dichloromethane-soluble fractions of the stack samples from all locations exhibited mutagenicity when tested in the plate incorporation assay. No significant response was observed with the acetone fraction. When tested with Kado's modification of the preincubation assay, the acetone-soluble fraction did exhibit mutagenic activity comparable to that of the other fractions when expressed in units of revertants per milligram of particular matter. Chemical analyses of one of the acetone-soluble fractions indicated that half of the mass was sulfuric acid while the remainder consisted of C, H and O. More than 30 peaks were detected in the high pressure liquid chromatogram of this fraction.

Although little mutagenic activity was detected in the polar ACE fraction of the diluted stack emissions samples with this single bioassay, in view of the large mass of this fraction, further investigation of the chemical composition and genotoxic activity of this fraction would be prudent.     

Mutagenicity evaluation of industrial sludge from common effluent treatment plant

Sludge from common effluent treatment plant (CETP) receiving effluents from textile industries at Mandia Road, Pali, was analyzed to assess the level of mutagenicity. Mutagenicity assay using Salmonella typhimurium tester strains TA 98 and TA 100 gave positive results, thus suggesting presence of genotoxic contaminants in the samples investigated. Further, mutagenic activity of chemical sludge was found to be lesser than that of biological sludge. This result is very surprising and unexpected as it is indicating that some mutagenic compounds are either being formed or certain promutagenic compounds are being converted into stable mutagenic metabolites during the biological treatment of the wastewater effluents. There have been no previous reports giving similar or contrary results. Most of the previous studies have reported effects of single combined sludge.     

Mutagenicity of ambient air pollutants collected near aluminum industries

Mutagenicity has been tested in air samples collected in the summer and in the winter near four Norwegian aluminum plants. The samples were separated into a particulate and a volatile fraction and tested for mutagenicity by a quantitative reversion assay which showed that the suspended particles were clearly mutagenic. The volatile part of the air pollutants were cytotoxic to the bacteria and showed only marginal mutagenicity. The particulate fractions were tested more extensively in the Ames Salmonella mutagenicity test, in two laboratories, using the strains TA 98 and TA 100 with and without enzymatic activation (S9). The mutagenicity was relatively high compared to particulate fractions from other areas with industry and dense traffic. The highest mutagenicity was found in TA 100 with enzymatic activation and the lowest in TA 100 without S9. The mutagenicity was influenced by wind speed and direction during sampling. The mutagenic activity was also determined in the nitroreductase deficient strains TA 98NR and TA 98/1.8DNP. A larger reduction in the activity was found compared to samples from other areas, thus indicating a difference in the sample composition.     

Genotoxicity of methyl parathion in short-term bacterial test systems

Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA-damage. Methyl parathion was mutagenic to S. typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes. In DNA repair tests, methyl parathion was effective in inducing damage to the S. typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms. Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion.     

Genotoxicity of methyl parathion in short‐term bacterial test systems

Abstract

Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA‐damage. Methyl parathion was mutagenic to S. typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes. In DNA repair tests, methyl parathion was effective in inducing damage to the S. typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms. Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion.     

Correlations between mutagenic activity of organic extracts of airborne particulate matter, NOx and sulphur dioxide in southern Germany: results of a two-year study

Goals, Scope and Background Among other substances, sulphur dioxide (SO2), nitric oxide (NO) and nitrogen dioxide (NO2) are parameters which are routinely measured to describe basic air quality. Organic extracts of airborne particulate matter contain mutagenic chemical compounds of different origins. The aim of the study was to find correlations between routine monitoring data and mutagenic activity of organic extracts of simultaneously drawn samples.Methods Specimens were collected over a period of two years at 8 sampling sites in south-west Germany. Simultaneously, concentrations of NO, NO2, and SO2 were measured on-line within the framework of the official air monitoring network of Baden-Württemberg, Germany. Dust samples were collected for biotesting using high volume air samplers equipped with glass fibre filters. After sampling was completed, filters were extracted and samples were prepared for biological testing. Mutagenic activity was tested by means of the plate incorporation assay (Ames test) using S. typhimurium TA98 and TA100 tester strains. During the first year of the study, all tests have been performed with and without metabolic activation. Additionally, a series of tests has been performed in parallel with TA98 and TA98NR.Results and Discussion Comparison of Ames test data obtained with and without metabolic activation indicates no statistically significant difference between both methods. Therefore, during the second year of the study, all tests have been performed without metabolic activation. Average yearly activities at the sampling sites were between 1 und 27 Revertants per m³ (Rev/m³). High activities were preferably found at congested sites (Karlsruhe, up to 95 Rev/m³). However, peak values of over 100 Rev/m³ were found in other places where pollution by traffic is significantly lower. The reason for these high level values is not evident. Tests performed using TA98NR tester strain indicate a significant share (average 31%) of compounds requiring activation by nitroreductase for mutagenic activity. Average mutagenic activity can be correlated to routine monitoring parameters. Comparison of averaged data for particular sampling sites indicates significant correlation between nitric oxide and mutagenic activity in TA98 (r²=0.90), while correlation between nitrogen dioxide (0.84) or sulphur dioxide (0.52) and mutagenic activity is weaker. For TA100, correlations are generally weaker than for TA98. Comparison of data for mutagenic activity and routine monitoring data of distant sites being sampled simultaneously shows parallel behaviour.Conclusions Results from this study show that mutagenic activity can be compared to seasonal and local variations of gaseous indicator air pollutants. Tester strain TA98 generally shows the best correlations. Although pollution by particle-bound mutagenic substances is significantly higher during the cold season than during summer on average, mutagenic activity of airborne dust is not a continuous effect. During winter, peak levels as well as low pollution periods can occur. Even during winter time mutagenic activity can reach very low levels typical for summertime. Comparison of results for distant sampling sites where samples have been collected simultaneously indicate that “classical” indicators of air pollution and bacterial mutagenicity of organic extracts from airborne particulate matter are influenced by connected effects. Seasonal trend of mutagenic activity, in particular, is similar to the concentrations of nitrogen oxide. NO is a strong indicator for vehicle exhaust gases. It is concluded that the average mutagenic activity at particular sites can be estimated using NO concentrations as an indicator.     

Investigating the sources of the mutagenic activity found in a river using the Salmonella assay and different water extraction procedures

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.     

QSAR modeling for predicting mutagenic toxicity of diverse chemicals for regulatory purposes

The safety assessment process of chemicals requires information on their mutagenic potential. The experimental determination of mutagenicity of a large number of chemicals is tedious and time and cost intensive, thus compelling for alternative methods. We have established local and global QSAR models for discriminating low and high mutagenic compounds and predicting their mutagenic activity in a quantitative manner in Salmonella typhimurium (TA) bacterial strains (TA98 and TA100). The decision treeboost (DTB)-based classification QSAR models discriminated among two categories with accuracies of >96% and the regression QSAR models precisely predicted the mutagenic activity of diverse chemicals yielding high correlations (R ²) between the experimental and model-predicted values in the respective training (>0.96) and test (>0.94) sets. The test set root mean squared error (RMSE) and mean absolute error (MAE) values emphasized the usefulness of the developed models for predicting new compounds. Relevant structural features of diverse chemicals that were responsible and influence the mutagenic activity were identified. The applicability domains of the developed models were defined. The developed models can be used as tools for screening new chemicals for their mutagenicity assessment for regulatory purpose.

     

Photochemical mutagen formation from aerobically treated night-soil effluent and the contributive substances

Effluents from a night-soil treatment plant, where night-soil was aerobically treated by an activated sludge process, were irradiated with a UV lamp excluding short wavelengths less than 300 nm as a model of exposure to sunlight and the mutagenicities of the ethylacetate extracts from the irradiated effluents were assayed using Salmonella typhimurium TA98. The extracts exhibited mutagenicity toward S. typhimurium TA98 in the absence of rat liver S9 fraction only when the effluents were fortified with nitrite ion (more than 6 ppm) by over aeration or by artificial addition of nitrite, indicating that a limiting factor for mutagen formation is nitrite ion concentration. Nine organic-N-containing compounds as models of the organic components in the effluent were also irradiated and direct-acting potent mutagens were found to be produced from such compounds having indole moiety as indole, oxindole, tryptophan and tryptamine.     

Seasonal variations and trends in concentrations of filter-collected polycyclic aromatic hydrocarbons (PAH) and mutagenic activity in the San Francisco Bay area

Air monitoring in the San Francisco Bay Area was carried out to measure outdoor community air concentrations of polycyclic aromatic hydrocarbons (PAH) and mutagenic activity (mutagenicity) in particulate organic matter (POM). Monitoring began in 1979 and is currently conducted at six stations. PAH and mutagenicity tests were performed on organic extracts prepared from high volume (hi-vol) filters composited every four months, by meterological season. PAH were determined by high pressure liquid chromatography (HPLC) with fluorescence and ultraviolet detection. Mutagenicity was measured in the Ames Salmonella bioassay using strain TA98 with and without metabolic activation. The nine-year mean concentration of benzo(a)pyrene (BaP) was 0.4 ng/m3. The mutagenicity of this amount of BaP accounted for only about 0.2% of the observed mutagenicity in POM and other measured PAH accounted for even less. Concentrations of PAH and mutagenicity were three to nine times higher during the winter than during other seasons. Year-to-year wintertime trends in several PAH were also seen. Early in the 1980s, winter concentrations of BaP and benzo (g,h,i)perylene increased. However since the mid-1980's, their concentrations have fallen. The decrease in PAH concentrations may be the result of an increasing proportion of vehicles with relatively low organic emissions. In contrast to PAH, mutagenicity did not show significantly year-to-year time trends.     

The chemical and biogenotoxic characterization of organic xenobiotics in aquatic sediment materials 1. The application and comparison of chemically non-specific and biogenotoxic methods

The aim of this work was to evaluate the Ames assay and mixed function oxidase (MFO)-Induct Test used in parallel with chemical group tests (ECD fingerprint and PAH estimation) for the characterization of the organic pollution of water sediment materials. Sediment materials were collected from “clean” and relatively heavily polluted locations in the Middle Adriatic Sea, and from some locations in continental Croatia polluted with wastewaters from different enterprises. Characterization of the organic extracts of the sediment materials investigated was performed chemically using UV spectrofluorometry for the determination polyaromatic hydrocarbons (PAH) and gas chromatography for the determination of volatile EC detector sensitive materials. Genotoxic analysis of the extracts was performed using the MFO-Induct Test and mutagenicity testing using the Standard Plate Incorporation Test as described by Maron and Ames with Salmonella typhimurium TA 98. Measurement of the BaPMO enzyme activity in the livers of carp treated i.p. with total extracts of the sediment investigated confirmed that the methanol extracts generally contained more inducing matter than the petroleum ether extracts. Ames assay showed that for all the samples following the elimination of the sulfur, there was an increase in the number of revertants in comparison to the control number, which indicates that the samples contained mutagenic substances. The larger doses of extracts generally demonstrated cytotoxicity, as evidenced by a reduced number of spontaneous revertants in the Salmonella/Microsome Test. Investigation of the correlation of the chemical parameters with the biological parameter showed that the induction of BaPMO exhibited a statistically significant correlation with the level of the ECD fingerprint of the petroleum ether sediment extract.     

Formation of mutagens by photolysis of amino acids in neutral aqueous solution containing nitrite or nitrate ion

Nine amino acids, i.e., alanine, threonine, cysteine, glutamic acid, arginine, proline, tryptophan, phenylalanine and tyrosine, were irradiated with UV light in water containing nitrite or nitrate ion under neutral conditions. The mutagenicities of the ether extracts and the residual water layers of the reaction mixtures were assayed with and without S-9 mix using $\text{Salmonella typhimurium}$ TA98 and TA100. Three aromatic amino acids, tryptophan, phenylalanine and tyrosine, were found to give direct-acting and frameshift mutagens by irradiation in aqueous nitrite solution. Among them, the ether extract of tryptophan exhibited the strongest mutagenicity toward TA98. In the case of irradiation in aqueous nitrate solution, only the ether extract of tryptophan exhibited weak mutagenicity toward TA98 without S-9 mix. The effects of nitrite concentration, irradiation time and pH on mutagen formation from tryptophan and some characteristics of the produced mutagens were examined.     

The efficiency of two standardized procedures for extracting mutagenic chemicals from soils

A major technical limitation to monitoring the fate of mutagenic compounds in soil is the lack of an established extraction procedure. This study was conducted to evaluate the efficiency of the blender extraction procedure for extracting benzo(a)pyrene (BAP) or 2-nitrofluorene (2NF) from a Weswood silt loam or a Bastrop clay loam soil. Extracted residues were evaluated using the Salmonella/microsome mutagenicity assay and high performance liquid chromatography (HPLC) to quantify recovery of the two chemicals from soil. In addition, a limited study using only the mutagenicity assay was conducted to compare the efficiency of the Blender and Soxhlet extraction procedures for recovering organic mutagens from both soils amended with a wood preserving bottom sediment waste.Extracted residues were dissolved in dimethylsulfoxide to concentrations that could be detected in the bioassay. Over all treatment levels and for both soils, the extraction efficiency of the blender procedure for BAP was greater than 80% for 12 of 16 treatments as measured using both bioassay and HPLC analysis; while, for 2NF the extraction efficiency was greater than 90% for all 15 treatments as measured by HPLC, and greater than 80% for twelve of 15 treatments as measured by bioassay. These results indicate that blender extraction provided efficient recovery of the pure compounds tested. When the efficiency of the blender and Soxhlet procedure for extracting the wood-preserving bottom sediment and waste amended soil were compared, significantly greater levels of mutagenic activity were detected in the fractions extracted using the blender extraction than were detected using the Soxhlet extraction.     

Changes in toxicity and genotoxicity of industrial sewage sludge samples containing nitro- and amino-aromatic compounds following treatment in bioreactors with different oxygen regimes

Goals, Scope and Background

From 2005, deposition of organic waste will be banned in Sweden. Likewise, in Germany and Austria, similar bans are being planned, and further countries will probably follow. Thus, there is a need to develop new methods and to refine established techniques for sludge management in the whole of the European Union. For this end, there is also an urgent need for appropriate ecotoxicological approaches to elucidate and assess the hazard potential of sewage sludge. Therefore, the present study was designed to assess the capacity of various established sludge treatment methods using different oxygen regimes to degrade recalcitrant nitro-substituted organic compounds and reduce their toxicity. Sewage sludge samples from a wastewater treatment plant in Sweden (Cambrex Karlskoga AB, industrial area Björkborn) receiving wastewater from industries manufacturing pharmaceutical substances, chemical intermediates and explosives were processed with different sludge treatment methods. Among other treatment methods, bioreactors (for anaerobic and aerobic sludge treatment) were used. In the present investigation, a battery ofin vitro bioassays was employed to compare the cytotoxic and genotoxic potentials of different fractions of sludge samples in order to elucidate whether the treatments were suitable to reduce the toxicity of the sludge.

Methods

In order to investigate the cytotoxicity of the extracts of treated and untreated sludge samples, the acute cytotoxicity test with the permanent cell line RTL-W1 was used. Genotoxicity was tested by means of the comet assay (single cell gel electro-phoresis) with RTL-W1cells, and mutagenicity was assessed with the Ames test using the Salmonella typhimurium strains TA98, TA98NR and TA100. Sludge toxicity was tested in different fractions of organic extracts produced by acetone and hexane extractions. The subsequent clean-up procedure (silica gel chro-matography and elution with hexane and dichloromethane) resulted in two fractions, a lipophilic hexane-fraction and a semi-lipophilic dichloromethane-fraction. For the genotoxicity and mutagenicity tests, these fractions were reunited at equal ratios.

Results and Discussion

The acute cytotoxicity test with RTL-Wl cells revealed a high cytotoxic potential for the semi-li-pophilic DM-fractions of all sludge samples with NR₅₀ values (= effective concentration for 50% cell death in the neutral red test) from 8.9 up to 20 mg sludge d.w./ml medium. A low cytotoxic potential for the hexane fractions of the untreated sludge samples (NR₅₀ 400 to < 400 mg sludge d.w./ml medium) was observed, whereas the hexane fractions of the treated sludge samples showed elevated cytotoxicity increasing further with treatment in the bioreactors. The comet assay indicated that three out of eight of the reunited fractions had a significant genotoxic potential. Whereas the genotoxic potential of one sample treated anaerobically was very high with an induction factor of 11.6, a similar sample (taken from the same anaerobic reactor four months later) and one untreated sample showed lower potentials. The samples treated in another anaerobic bioreactor as well as the samples treated aerobically showed no genotoxic potential. Results indicate that aerobic treatment was basically adequate for reducing the genotoxicity of the sludge, whereas anaerobic treatment was only partly useful for reduction of genotoxicity. The Ames test revealed a very high mutagenic potential for the reunited fractions of the untreated sludge samples with strain TA98 (maximum induction factors (IF_max up to 45) and a relatively high potential for one of the samples treated aerobically (S2, IF_max = 18 (TA98, S9-)), thus documenting the suitability of both anaerobic and aerobic treatments to reduce the mutagenicity of the samples, however, with the aerobic treatment being less effective. Conclusions. Overall, none of the microbiological treatments for wastewater sludge in bioreactors was found to be ideal for general toxicity reduction of the sludge samples. Whereas cytotoxicity of the sludge increased or levelled off in most cases following either treatment, genotoxicity both increased or decreased after anaerobic treatment, depending on the specific sample. However, mutagenicity could generally be reduced by anaerobic treatment and, to a lesser degree, by aerobic treatment. Recommendationsand Perspectives. The complex modification of the diverse damage potentials of sludge sample extracts by use of anin vitro biotest battery following treatment for toxicity reduction in bioreactors showed that considerations of different toxicological endpoints is essential for an adequate hazard assessment. Whereas in the case of cytotoxicity reduction, the reactors proved ineffective, mutagenicity could be reduced significantly at least in some cases in this case study.

     

Chemical and biological profiles of sediments as indicators of sources of contamination in Hamilton Harbour. Part II: bioassay-directed fractionation using the Ames Salmonella/microsome assay

Bottom sediment and suspended sediment samples from Hamilton Harbour (western Lake Ontario) and from a major tributary were profiled using a bioassay-directed fractionation approach. Sample extracts were fractionated using an alumina/Sephadex gel clean-up procedure to afford non-polar aromatic fractions which were characterized using chemical analyses and the Ames/microsome bacterial assay in Salmonella typhimurium strains YG1025 with the addition of oxidative metabolism (S9), and YG1024 without S9. Non-polar aromatic fractions of selected samples were separated by normal phase HPLC into 1-min fractions which were subjected to bioassay analyses. The bioassays using strain YG1025+S9, a TA100-type strain, were performed to assess genotoxicity arising from the presence of polycyclic aromatic hydrocarbons (PAH). Fractions which exhibited mutagenic activity contained PAH with molecular masses of 252, 276 and 278 amu; these fractions contained over 80% of the genotoxicity attributable to PAH. Individual compounds identified using Gas Chromatography-Mass Spectrometry analyses in these active fractions included benzo[a]pyrene, indeno[cd]pyrene and dibenz[a,h]anthracene. The YG1025+S9 mutagenic activity profiles were similar for all samples. Mutagenic activity profiles generated using strain YG1024-S9, a TA98-type strain sensitive to compounds characteristic of mobile source emissions, were very different. The mutagenic activities in strain YG1024-S9 were greatest for harbour-suspended sediment samples collected from sites impacted by a major tributary. Suspended sediments collected near areas known to contain high levels of coal tar-contamination in the bottom sediments contained higher levels of genotoxic PAH than suspended sediments collected from other areas of the harbour.     

Application of semipermeable membrane device (SPMD) to assess air genotoxicity in an occupational environment

Semipermeable membrane device (SPMD) is a passive sampler that sequesters lipophilic contaminants, mimicking the bioconcentration in the fatty tissue of organisms. This study was designed to assess the use of SPMD and biological tests (Comet assay and Ames test) for air monitoring. For this purpose an occupational environment with expected polycyclic aromatic hydrocarbons (PAHs) contamination (coke plant) was selected for a case study. The SPMDs were deployed in five occupational contaminated sites and in a control site. The SPMD dialysates were chemically analysed and examined for in vitro DNA-damaging activity in human cells (Jurkat) by Comet assay and for mutagenicity with the Ames test (TA98 strain, w/o S9). Total suspended particulates were also collected and analysed (GC–MS). No biological effect of SPMD extract was revealed in the control site. On the other hand, air samples collected with SPMDs within the coke plant showed variable degrees of genotoxic and mutagenic activity. The highest effects were associated with the highest PAH level recovered in the SPMDs extracts and in particulate samples.Results obtained support the sensitivity of biological tests associated to SPMD sampling for evaluating the health risk of potentially contaminated work environments highlighting the usefulness of SPMDs for environmental air quality monitoring.