Detection and average content levels of carcinogenic and mutagenic compounds from the particulates on diesel and gasoline engine mufflers

Carcinogenic and mutagenic compounds, which were extracted from the particulates that adhered to inner surfaces of diesel and gasoline engine mufflers, were quantified by the series method of Soxhlet extraction, liquid-liquid partition, thin-layer chromatography, and spectrofluorometry. Mutagenic activity of their neutral and acidic fractions was tested in the improved Ames assay by the preincubation method with Salmonella typhimurium TA98 in the presence and absence of metabolic activation system (S-9 mix). The average content levels (μg/g tar) of polycyclic aromatic hydrocarbons from gasoline engine cars were greater than those from diesel engine vehicles. However, the levels of nitro derivatives of PAHs and polycyclic quinones from the diesel engines were greater than from the gasoline engines. Mutagenic activity of the diesel acidic fraction was the highest among the diesel and gasoline fractions, and was significantly higher in the absence of the S-9 mix. Furthermore, the relative value (R_{c = 0}) of infrared absorption of carbonyl stretching vibration to that of methylene asymmetric stretching vibration of the diesel acidic fraction was the highest among the diesel and gasoline fractions. These results strongly suggest that highly direct-acting mutagens in the acidic fraction are at higher levels in diesel emission particulates than those from gasoline, and that these mutagens are carboxylic acid, aldehyde, and alcohol derivatives of PAHs and NPAHs.     

DNA-binding studies with diesel exhaust particle extract

This study examines whether chemical components from diesel exhaust particulates react with DNA to form covalently bound adducts. Experiments in this report describe the in vitro reaction of purified DNA with a dichloromethane extract of diesel exhaust particulates in the absence or presence of enzyme activation by rat liver microsomes. The reactivity of the particle extract was compared to that of benzo[a]pyrene metabolites using low temperature fluorescence techniques which detect small quantities of polycyclic aromatic compounds bound to DNA. Incubation of DNA with the particle extract in the presence of microsomal enzymes produced no detectable fluorescent adducts in contrast to model experiments using benzo[a]pyrene. However, addition of the particle extract to incubation mixtures containing benzo[a]pyrene markedly decreased formation of benzo[a]pyrene-DNA adducts because the particle extract inhibits microsomal enzymes which activate benzo[a]pyrene and other polycyclic aromatic hydrocarbons. In the absence of microsomal enzymes, fluorescent material was detected in DNA exposed to high concentrations of the particle extract, but probably not as a result of covalent binding because the mutagenic activity of the particle extract remained unchanged during prolonged incubation with DNA. This stability is in contrast to the rapid decrease in mutagenic activity of benzo[a]pyrene-4,5-oxide during incubation with DNA. Thus, direct mutation of bacteria by the particle extract may require activation by bacterial enzymes as is known to occur with nitroaromatic compounds.     

Identification and quantitative confirmation of dinitropyrenes and 3-nitrobenzanthrone as major mutagens in contaminated sediments

Polar fractions of a sediment extract of the industrial area of Bitterfeld, Germany, have been subjected for effect-directed identification of mutagens using the Ames fluctuation assay with TA98. Mutagenicity could be well recovered in several secondary and tertiary fractions. Dinitropyrenes and 3-nitrobenzanthrone could be confirmed to contribute great shares of the observed mutagenicity. In addition, a multitude of polar polycyclic aromatic compounds has been tentatively identified in mutagenic fractions including nitro-PAHs, azaarenes, ketones, quinones, hydroxy-compounds, lactones and carboxylic acids although their contribution to mutagenicity could not be quantified due to a lack of standards. Diagnostic Salmonella strains YG1024 and YG1041 were applied to confirm the contribution of nitro-aromatic compounds. We suggest the inclusion of dinitropyrenes and 3-nitrobenzanthrone into sediment monitoring in order to minimize the mutagenic risk to aquatic organisms and to human health.     

Mutagenic activity of diesel emission particulate extracts and isolation of the mutagenic fractions

Mutagenic activity in diesel emission particulate extracts was detected by the Salmonella typhimurium/microsome assay. Direct-acting mutagens as well as promutagens requiring metabolic activation were detected. The extracts were fractionated into acidic, basic, and neutral fractions, and the neutral fraction was chromatographed into seven subfractions. Differences in the mutagenic potency of these fractions and subfractions were determined by the Salmonella assay. Fractions containing as yet unidentified compounds, but not polynuclear aromatic hydrocarbons, were found to make a major contribution to mutagenic activity of the extracts.     

Genotoxicity of syrian hamster lung cells treated in vivo with diesel exhaust particulates

Polycyclic aromatic hydrocarbons extracted and concentrated from diesel exhaust particulates have been shown to be mutagenic and carcinogenic, but attempts to induce pulmonary tumors through chronic inhalation of diesel exhaust by experimental animals have failed. We have attempted to resolve this incongruity by measuring chromosomal damage in lung tissue of chronically exposed hamsters, using the highly sensitive test for genotoxic chemical agents, sister chromatid exchange (SCE) analysis. To determine the degree of responsiveness of the test system to both diesel exhaust particulates and benzo(a)pyrene (BaP), these agents were instilled intratracheally into anesthetized hamsters as suspensions in 0.25 ml volumes of Hank's balanced salt solution (HBSS). Lung tissues from these animals were subsequently cultured in vitro and chromosomes from the resulting cell divisions were scored for exchanges of chromatin between sister chromatids. Control animals, treated weekly with 0.25 ml of BSS for 10 weeks, showed an average value of 12 SCE's per cell, while animals treated weekly with 200 ng BaP over a 10-week period showed an average of 17 SCE's per cell. HBSS, given as a single treatment, also produced an average of 12 SCE's per cell in control animals, but animals treated with a single instillation of 12.5 μg BaP showed an average SCE value of 19. These data confirmed that the procarcinogen BaP can be metabolically activated by lung cells in vivo and also demonstrated the efficacy of using this technical approach to study the effect of chemical mutagens that enter the lungs. Diesel exhaust particulates, administered in a range from 0 to 20 mg per hamster over a 24 h exposure period, produced a linear SCE dose-response ranging from 12 to 26 SCE's per metaphase. This curve suggested that a concentration of 3 mg of diesel particulates per hamster would not produce a statistically significant increase in SCE's above control values. One group of 8 hamsters, chronically exposed to diesel exhaust particulates for 3 months showed an average of 12 SCE's per cell. This was equivalent to a set of 5 control animals which also showed an average of 12 SCE's per cell. Although the scope of this study was limited, the data demonstrated that diesel exhaust particulates can induce genotoxic damage but a 3-month exposure to 6 mg/m³ of diesel exhaust particulates was insufficient to produce measurable mutagenic changes in lung cells. This negative response is consistent with the results from other studies in which similar exposures failed to produce pulmonary tumors.     

Mutagenicity, genotoxicity and carcinogenic risk assessment of indoor dust from three major cities around the Pearl River Delta

The mutagenicity and genotoxicity of workplace dust including commercial office, secondary school, shopping mall, hospital, electronics factory and manufacturing plant in Hong Kong and settled house dust from Hong Kong, Shenzhen and Guangzhou were measured. Results indicated that indoor dust contained both frameshift and base pair substitution mutagens. Dust from manufacturing plant showed highest mutagenic potency on TA98±S9 and TA100±S9 activation, whereas, electronics factory showed highest genotoxicity with and without S9 activation. TA100 (-S9) mutagenic potency was significantly correlated with genotoxicity expressed as SOSIP (-S9) of workplace dust (r(2)=0.37, p<0.01). The total PAHs concentration of settled house dust from PRD ranged from 1.63 to 29.2μg/g. Linear regression analyses indicated that the PAHs likely accounted for about 45% of the TA98 with S9 mutagenic activity of workplace dust. TA98 (-S9) mutagenicity (r(2)=0.27, p<0.05) and SOSIP (-S9) of house dust (r(2)=0.41, p<0.01) were both significantly correlated with the number of inhabitants in the house. To achieve a more accurate cancer risk assessment, the oral bioaccessibility of B(a)A, Chry, B(b+k)F, B(a)P, D(ah)A and I(cd)P in different dust ranging from 1.3% to 17% was taken into account. Risk assessments indicated that about 26% of house dust samples resulted in unacceptable cancer risk (>1×10(-6)) for preschool children.     

Evaluation of the mutagenic potential of some compounds using Salmonella typhimurium

The mutagenicity of sodium nitrite, potassium chromate, O-tolidine, and smoke condensate from tobacco was shown on Salmonella typhimurium TA 1535. Sodium nitrite was definitely mutagenic on TA 1535 without S-9 mix. Potassium chromate without S-9 mix was also mutagenic on TA 1535. However, addition of S-9 mix resulted in a complete loss of potassium chromate mutagenicity. O-tolidine with and without S-9 mix elicited a very weak and variable mutagenic effect for strain TA 1535. Smoke condensate from pipe tobacco without S-9 mix elicited a weak mutagenic effect, whereas with S-9 mix it showed no mutagenicity on TA 1535.     

Diesel particulate extracts in bacterial test systems

The Ames bacterial mutagenicity test system was used to evaluate parameters which may affect the mutagenic activity of diesel particulate extracts. The optimal extraction conditions, extractability of mutagens by simulated biological fluids and the effect of collection method were investigated. The role of solvent was examined by extracting diesel particles with methanol, acetone, cyclohexane, ethyl acetate, n-hexane, dichloromethane, benzene and a benzene-ethanol mixture. Of these, the dichloromethane extract exhibited the highest activity in the Ames test, although methanol yielded the largest extractable mass. Diesel particles were also extracted by dimethyl sulfoxide (DMSO) and four other simulated biological fluids for 48 h at 25, 37, and 45°C to study the effects of temperature. The mutagenic activity of the DMSO extract began to decline at temperatures higher than 37°C after 8 h of incubation. Fetal calf serum was the only simulated biological fluid which eluted mutagenic activity from the particles. No activity was detected in the 0.5% bovine serum albumin, simulated lung surfactant and saline extracts. Diesel particles collected by electrostatic precipitation (ESP) and filtration were studied. The mutagenic activities of both extracts were comparable when expressed as revertants per mg of particle. After the extracts were separated into nine fractions by a solvent partitioning scheme, the majority of the activity was found in the neutral-nonpolar II, neutral polar, strong acid and weak acid fractions. The acid salt fraction from the ESP sample was inactive. These results demonstrate that differences in the extraction conditions can result in differences in the mutagenic activity of diesel particulate extract. Since the mutagens in the extracts are not readily extractable by simulated biological fluids, the question of bioavailability of mutagens in diesel particles must be considered in the final assessment of their potential effects in biological systems and organisms.     

Deposition and biological availability of diesel particles and their associated mutagenic chemicals

To estimate the human health risk of inhaled diesel particles, it is necessary to know their deposition and retention in the respiratory tract and the rate of dissociation of mutagenic compounds associated with the particles. The deposition of a chain aggregate aerosol of ⁶⁷Ga₂O₃ with size and shape characteristics similar to diesel exhaust particles has been evaluated using Beagle dogs. Approximately one-third of the inhaled activity is deposited in the respiratory tract with most of the particles deposited in the lung. The mutagenic activity present in dichloromethane, dog serum, dog lung lavage fluid, saline, dipalmitoyl lecithin (DPL) and albumin following incubation of these fluids with diesel exhaust particles was determined in the Ames Salmonella system. As observed by other investigators, large quantities of mutagenic activity were removed by dichloromethane. A very small amount of mutagenic activity was removed by the serum and lavage fluid over a 3-day incubation period. No activity was detected following elution with the other solvents. The finding that minimal mutagenic activity could be demonstrated in the biological media following incubation with diesel exhaust particles may be due to a lack of removal of mutagens from the particles or an inactivation of removed mutagens by protein binding or other processes.     

Microbial mutagenicity and toxicity of newly synthesized heterocyclic N-oxides

Newly synthesized heterocyclic N-oxides were tested for their mutagenicity using the Ames Test. DX1 was potentially mutagenic in Salmonella typhimurium TA 100 and TA 98 with and without the S-9 mixture. WO-25 and WO-20, which are structurally related to DX1 did not show genetic change in the strains used. The antibiotic activity of these chemicals was also tested using gram negative and gram positive bacteria. The mutagenic chemical DX1 had more killing effect in gram positive bacteria than WO-25 and WO-20.     

Evaluation of the water genotoxicity from Santos Estuary (Brazil) in relation to the sediment contamination and effluent discharges

The genotoxic activity of water samples collected in 9 different sites within the area of the Santos estuary was preliminary evaluated, and related to previous data on the genotoxicity of sediments and the contents of PAHs in both water and sediment samples. The liquid discharge of a steel mill (coke plant), known to be mutagenic, was chemically analyzed to determine its PAH content. For the water evaluation we employed the Salmonella/microsome assay with the strains TA98 and TA100 with and without S9 mix in the plate incorporation method. The water was filtered with an AP20 membrane before being extracted with XAD4 at natural and acidic pH. The industrial effluent was filtered in 0.45 microm membranes before being extracted with the liquid/liquid method. Both membranes containing the particulate material were extracted using ultrasonication. PAHs were found associated with the suspended particles present in the industrial effluent in accordance with mutagenicity data previously reported. In relation to the estuarine waters, sites 1 and 5 presented low levels of mutagenic activity only in the filtered water (liquid fraction) extracts. At site 3, both the filtered water and particulate solids presented also low mutagenicity. Results show that the mutagenic activity observed in water could not be directly related to the genotoxic activity and PAHs contents of the bottom sediments.     

Soil mutagenicity as a strategy to evaluate environmental and health risks in a contaminated area

Soil can be a storage place and source of pollutants for interfacial environments. This study looked at a site contaminated with wood preservatives as a source of mutagens, defined routes and extent of the dispersion of these contaminants by particle remobilization and atmospheric deposition, considering an evaluation of risk to human health by quantifying mutagenic risk. Soil sampling sites were chosen at gradually increasing distances (150, 500 and 1700 m) from SI (industrial area pool) and indoor dust (pool in an area at risk at 385 m and at 1700 m). Mutagenesis was evaluated in the Salmonella/microsome assay, TA98, TA97a and TA100 strains with and without S9 mix, YGs strains 1041, 1042 and 1024 for nitrocompounds. Acid extracts were analyzed to define the effects of metals and organics for polycyclic aromatic hydrocarbons (PAHs) and nitroderivates, besides concentrations of these compounds and pentachlorophenol (PCP). Risk to human health was obtained from the relation between the quantified potential of mutagenic risk and estimated soil ingestion for children according to USEPA. Metal concentrations showed a gradient of responses with As, Cr and Cu (total metal) or Cr and Cu (fraction available) higher for SI. However, mutagenic effects of the mixtures did not show this grading. Site SR1700, without a response, was characterized as a reference. In organic extracts, the mutagenesis responses showed the mobility of these compounds from the source. In the surrounding area, a smaller pattern similar to SI was observed at SR150, and at the other sites elevated values of direct mutagenesis at SR500 and diminished effects at SR1700. Tests with YG strains indicated that nitrated compounds have a significant effect on the direct mutagenesis found, except SR500. The investigation of indoor dust in the surrounding area enabled confirmation of the particle resuspension route and atmospheric deposition, showing responses in mutagenicity biomarkers, PAH concentrations and PCP dosage similar to SI. The range of values obtained, considering the soil masses needed to induce mutagenicity was 0.02 to 0.33 g, indicating a high risk associated with human populations exposed, since these values found surpass the standard estimate of 200 mg/day of rate of soil ingestion for children according to USEPA. The study showed that it is essential to evaluate the extent of contamination from the soil to delimit remedial measures and avoid damage to the ecological balance and to human health.     

Composition, sources, and potential toxicological significance of PAHs in the surface sediments of the Meiliang Bay, Taihu Lake, China

Twenty-five surface sediment samples were collected from Meiliang Bay, Taihu Lake, China, in 2003. The concentrations of 16 polycyclic aromatic hydrocarbons (PAHs), identified as priority pollutants by the USEPA, were determined by gas chromatography equipped with a mass spectrometry detector (GC-MS). Total concentrations of the PAHs ranged from 1207 to 4754 ng/g dry weight. Sediment samples with the highest PAH concentrations were from the northern site of the bay, which is in proximity to the incoming PAH source; the PAH levels in the southern part were relatively low. The observed PAH levels were higher than those in river sediments in China but were lower than those found in sediments of urban areas and harbors. According to the observed molecular indices, PAHs originated largely from the high-temperature pyrolytic process, whereas the petrogenic process was more commonly responsible for PAH contamination in harbors. A good correlation existed between the benzo[a]pyrene level and the total PAH concentration (r=0.97), making benzo[a]pyrene a potential molecular marker for PAH pollution. According to the numerical effect-based sediment quality guideline (SQGs) of the United States, the levels of PAHs at most studied sites in Meiliang Bay, except some sites in the northern part of the bay, should not exert adverse biological effects. In the northern part of the bay, the PAH levels at sites 21 and 22 exceed the effects range low (ERL) and could thus cause acute biological impairments, in comparison with the sediment quality guidelines. The total PAH levels were expressed as the B[a]P toxicity equivalents (TEQs(carc)) and compared to the contaminated sediments from Guba Pechenga, Barents Sea, Russia.     

Mutagenic properties of PM2.5 urban pollution in the Northern Italy: The nitro-compounds contribution

PM2.5 is the breathable fraction of the particulate matter and some adverse health effects, such as respiratory functionality, cardiological diseases and cancer, can be in some measure attributable to this risk factor exposure. Some of the most carcinogen compounds transported by PM2.5 are nitro-compounds. In this study, a strengthened in vitro bioassay — able to predict the mutagenic/carcinogenic activity of the environmental mixtures — was conducted on PM2.5 organic extracts to define the nitro-compounds burden. PM2.5 air pollution was daily monitored, during 2006, in three cities located in the Northern part of Italy (Torino, Pavia and Verona) and the mutagenic properties of the PM2.5 organic extracts were assessed with the Ames test. The bacterial used in this study were three Salmonella typhimurium strains: TA98, nitroreductase-less mutant TA98NR and YG1021 carrying a nitroreductase-producing plasmid. The annual PM2.5 mean level measured in Torino was 46.5 (± 31.6) μg/m³, in Pavia 34.8 (± 25.1) μg/m³, and in Verona 37.3 (± 27.8) μg/m³, while the mutagenicity expressed as TA98 net reverants/m³ was 28.0 (± 22.1), 28.3 (± 24.9), and 34.2 (± 30.9) respectively. Monthly pool bioassays, conducted with the three different strains, showed a greater mutagenic response of the YG1021 in each city. The relationship among the mutagenic answers for YG1021:TA98:TA98NR was about 6:3:1 (p < 0.001). Over nitroreductase activity enhanced the response of 2.2, 2.0 and 1.7 times for Torino, Pavia, and Verona (ANOVA Torino p < 0.05) respectively. Without nitroreductase activity the genotoxicity was limited. These biological findings are able to describe a relevant role played by the nitro compounds in the mutagenic properties of the urban PM2.5 in the Padana plain; moreover the bacterial nitroreductase plays a predominant role in DNA interaction primarily for Torino PM2.5 extracts.     

Diesel soot: Mutation measurements in bacterial and human cells

Both hot pipe and dilution chamber samples of the exhaust from a diesel (Oldsmobile 350) engine have been collected, extracted with methylene chloride and those extracts have been tested for mutagenicity in forward mutation assays in human lymphoblasts and S. typhimurium. In the absence of a metabolic activation system, the extract was significantly mutagenic to the bacteria in the range of 0 to 30 μg/ml, but introduced no mutations in human cells at concentrations up to 200 μg/ml under the same conditions of assay medium. However, when assayed in the presence of a postmitochondrial supernatant derived from rat liver, the the soot extracts were significantly mutagenic to both bacteria and human cells in the range of 50–100 μg/ml. Fractionation of the soot extract on the basis of polarity by sequential elution from a silicic acid column permitted concentration of the mutagenic activity in the alkane/toluene eluate, as determined by bacterial assays. Preliminary characterization of this fraction and preliminary studies of pure compounds leads us to suspect the alkyl substituted phenanthrenes as representing at least a significant fraction of the mutagenic activity of this alkane/toluene eluate.     

Carcinogenic potential, levels and sources of polycyclic aromatic hydrocarbon mixtures in indoor and outdoor environments and their implications for air quality standards

Both the World Health Organization and the UK Expert Panel on Air Quality Standards (EPAQS) have considered benzo(a)pyrene (BaP) as a marker of the carcinogenic potency of the polycyclic aromatic hydrocarbons (PAH) mixture, when recommending their respective guidelines for PAHs in outdoor air. The aim of this research is to compare the concentrations and relative abundance of individual PAH and their contribution to the overall carcinogenic potential of the PAH mixture in indoor and outdoor environments to assess the suitability of the UK air quality standard derived for outdoor air for use as a guideline for indoor environments. Samples were collected onto filters using active sampling in different indoor and outdoor microenvironments. The ratio of individual compounds to BaP, the BaP equivalent concentrations and the percentage contribution of each individual compound to the total carcinogenic potential of the PAH mixture were calculated. Mean concentrations were generally lower indoors (BaP=0.10 ng/m(3)) than outdoors (BaP=0.19 ng/m(3)), with the exception of indoor environments with wood burners (BaP=2.4 ng/m(3)) or ETS (BaP=0.6 ng/m(3)). The ratio of individual PAHs to BaP showed no significant differences between indoors (e.g. DahA/BaP=0.27) and outdoors (DahA/BaP=0.31). The relative contribution of BaP to the PAH overall carcinogenic potency is similar indoors (49%), outdoors (54%) and in the smelter environment (48%) used by EPAQS to derive the UK Air Quality Standard for ambient air. These results suggest the suitability of BaP as a marker for the carcinogenic potential of the PAH mixture irrespective of the environment. Despite small differences in PAH mixture composition indoors and outdoors, the level of protection afforded by the present EPAQS standard is likely to be similar whether it is applied to indoor or outdoor air.     

河口滨岸悬浮颗粒物中多环芳烃分布与风险评价

通过收集长江口滨岸13个典型采样点上覆水中的悬浮颗粒物,分析了悬浮颗粒中多环芳烃(PAHs)的含量水平,探讨了PAHs的来源,并进行了生态风险评价。研究结果显示,EPA14种优控PAHs的总量在600~12 308 ng/g 之间,平均值为5 373 ng/g,其组成主要以3环和4环PAH为主。受附近陆源输入的影响,顾路采样点的PAHs含量最高,此外,临近城市排污、滨岸工业开发区及河道排污口的采样点PAHs含量也较高,如石洞口、金山、白茆、浏河口等。结合PAHs不同环数的相对丰度与同分异构体荧蒽/芘、芘/苯并［a］蒽比值,初步推断出人类油污染及矿物燃料的不完全燃烧是悬浮颗粒物中PAHs的主要来源。此外,参照有关环境质量标准,发现悬浮颗粒物中萘、菲、芴、蒽/苯并［b］荧蒽和苯并［k］荧蒽等化合物已经产生不同程度的生物影响效应。     

Human dietary exposure to polycyclic aromatic hydrocarbons: Results of the second French Total Diet Study

In the frame of the second French Total Diet Study (TDS), the 15 + 1 EU priority polycyclic aromatics hydrocarbons (PAHs) were analyzed in 725 foodstuffs habitually consumed by the French population, using gas chromatography coupled to tandem mass spectrometry, after pressurized liquid extraction and purification on PS-DVB stationary phase. The highest PAH concentrations recovered in foodstuffs corresponded to the following contributors: chrysene (25.7%), benzo[b]fluoranthene (15.0%) and benz[a]anthracene (9.0%) whereas the lowest concentrations were those of dibenz[a,h]anthracene, 5 methylchrysene and dibenzo[a,h]pyrene (below 2.0%). By food groups, the current highest levels of total PAH were detected in mollusks and crustaceans, followed by the different oil based products. To estimate French population's exposure, contamination data were combined with national individual food consumption data. Mean daily exposure to the sum of benzo[a]pyrene, benz[a]anthracene, chrysene and benzo[b]fluoranthene (PAH4) was estimated to be 1.48 ng/kg bw/day in adults and 2.26 ng/kg bw/day in children. The main contributors to PAH exposure for adults are fats, bread and dried bread products followed by crustaceans and mollusks. The margin of exposure (MOE) approach indicates that exposure to PAHs through food is not a major health problem for French consumers.     

Salmonella/microsome mutagenicity assays of exhaust from diesel and gasoline powered motor vehicles

Motor vehicle exhaust from prechamber injection diesel and gasoline powered passenger cars, sampled during US FTP 1973 test cycles and comprising both particulate matter and compounds condensable at ambient temperature, has been assayed for mutagenicity in the Salmonella/microsome test. Mutagenic components were to a large extent active in the absence of the mammalian microsomal preparation. The mutagenicity of both particulate matter and condensate from diesel exhaust and condensate from gasoline exhaust was decreased in the presence of the microsomal preparation whereas the mutagenicity of particulate matter from gasoline exhaust was enhanced by microsomal activation. A comparison between the investigated diesel and gasoline exhaust samples shows that the mutagenic effect in the Salmonella test of the diesel exhaust is more than ten times higher than that of the gasoline exhaust. Fractionation with respect to polarity indicates that the mutagenic components mainly are distributed in neutral aliphatic, aromatic, and oxygenated fractions. Tests for mutagenic monofunctional nitroarenes by an anaerobic assay indicate that such compounds at most are marginally present in the exhaust samples as compared with their presence in airborne particulate matter collected in an urban environment.     

Habitat function of agricultural soils as affected by heavy metals and polycyclic aromatic hydrocarbons contamination

Ecotoxic activity of soils polluted with polycyclic aromatic hydrocarbon (PAH) and heavy metals (HM) was evaluated in pot and laboratory experiments. Plants and soil microorganisms were chosen as test organisms and six different soil materials were used in the study. The applied levels of HM and PAH were aimed to reflect environmental conditions in the "worst case" situation. Zn(2+), Pb(2+) and Cd(2+) were introduced to the soils as an aqueous solution of the mixture of salts at the concentrations corresponding to 1000, 500 and 3 mg kg(-1), respectively. Mixture of four PAH compounds (flourene, anthracene, pyrene and chrysene) as a CH(2)Cl(2) solution was applied at levels of 10-100 mg summation operator 4PAH kg(-1). Population and activity of soil microflora was evaluated as measured of total bacteria counts, intensity of respiration and enzyme activity (dehydrogenases and phosphatases). Effect on plants was evaluated on the base of the growth (plant at an early stage of their development) and yield (mature plant) measurements. The results indicate that combined effect of PAH and heavy metals on soil microorganisms activity and on some plants at an early stage of their development can be stronger than in soils amended with HM or PAH separately. Reaction of tested organisms was related to soil properties, PAH concentration, time and plant species. Mature plants (maize) were insensitive to the applied levels of both group of contaminants.