Effect of soil pollution with metallic lead pellets on lead bioaccumulation and organ/body weight alterations in small mammals

Using small mammals as bioindicators, the bioavailable status and ecotoxicity of lead was investigated in an acidic sandy soil environment polluted with metallic lead pellets from shotgun ammunition. Average concentrations of lead in kidney, liver and bone tissue of wood mice (Apodemus sylvaticus), bank voles (Clethrionomys glareolus), and shrews (Sorex araneus) were strongly elevated, compared to tissue levels of conspecifics collected from an adjacent unpolluted area. All shrews and some bank voles collected from the shooting range exceeded the critical renal Pb concentration of 25 g/g dry weight, considered diagnostic of lead intoxication in mammals. The geometric mean renal Pb concentration in shrews was 270 g/g dry weight, with an upper range of more than 1,000 g/g. The population of shrews and bank voles from the shooting range also showed a significantly increased average relative kidney weight (kidneyto-body weight ratio), which is indicative of lead poisoning. The results suggest that metallic lead pellets deposited in an acidic sandy soil are transformed to a chemical form of lead, probably Pb²⁺, which is toxic to organisms and which can readily enter the terrestrial food chain.     

Heavy-Metal Concentrations in Small Mammals from a Diffusely Polluted Floodplain: Importance of Species- and Location-Specific Characteristics

The soil of several floodplain areas along large European rivers shows increased levels of heavy metals as a relict from past sedimentation of contaminants. These levels may pose risks of accumulation in food webs and toxicologic effects on flora and fauna. However, for floodplains, data on heavy-metal concentrations in vertebrates are scarce. Moreover, these environments are characterised by periodical flooding cycles influencing ecologic processes and patterns. To investigate whether the suggested differences in accumulation risks for insectivores and carnivores, omnivores, and herbivores are reflected in the actual heavy-metal concentrations in the species, we measured the current levels of Zn, Cu, Pb, and Cd in 199 specimens of 7 small mammal species (voles, mice, and shrews) and in their habitats in a diffusely polluted floodplain. The highest metal concentrations were found in the insectivorous and carnivorous shrew, Sorex araneus. Significant differences between the other shrew species, Crocidura russula, and the vole and mouse species was only found for Cd. The Cu concentration in Clethrionomys glareolus, however, was significantly higher than in several other vole and mouse species. To explain the metal concentrations found in the specimens, we related them to environmental variables at the trapping locations and to certain characteristics of the mammals. Variables taken into account were soil total and CaCl₂-extractable metal concentrations at the trapping locations; whether locations were flooded or nonflooded; the trapping season; and the life stage; sex; and fresh weight of the specimens. Correlations between body and soil concentrations and location or specimen characteristics were weak. Therefore; we assumed that exposure of small mammals to heavy-metal contamination in floodplains is significantly influenced by exposure time, which is age related, as well as by dispersal and changes in foraging and feeding patterns under influence of periodic flooding.     

Accumulation of Lead,Zinc, and Cadmium in a Wild Population of <Emphasis Type="Italic">Clethrionomys glareolus</Emphasis> from an Abandoned Lead Mine

Lead, zinc, and cadmium were determined in a range of tissues from wild populations of bank voles (Clethrionomys glareolus) trapped on an abandoned metalliferous mine site and a reference site. Estimated dietary intakes indicated that animals were exposed to elevated levels of all three metals at the mine site, and this was generally reflected in metal residues in body tissues. Lead concentrations were significantly higher in all tissues of animals from the mine compared to the reference site, while Cd was higher only in the kidney. There was evidence of age-accumulation (using total body weight as an index of age) of Cd in both the liver and kidney of mine site animals but no evidence of such accumulation of lead in bone. In contrast to Cd and Pb, Zn was lower in the tissues of mine site animals compared to the reference site. Based on critical tissue concentrations, the ecotoxicological risk to a wild population of bank voles (Clethrionomys glareolus), associated with total substrate levels of 1 μg g⁻¹ dry weight Cd and 700 μg g⁻¹ dry weight Zn at this mine site is negligible, but that associated with 4000 μg g⁻¹ dry weight Pb may be considered significant. Received: 28 January 2002/Accepted: 29 July 2002     

Aromatic DNA adducts in coke-oven workers, in relation to exposure, lifestyle and genetic polymorphism of metabolic enzymes

Objective: This study investigates the effect of multiple factors, including exposure to polycyclic aromatic hydrocarbons (PAHs), lifestyle, genetic polymorphism of cytochrome P450 (CYP)1A1, glutathione transferase (GST)M1, GSTP1, N-acetyltransferase (NAT)2 and gene p53, as well as any family history of cancer, on DNA adduct levels in coke-oven workers. Methods: Sixty-five coke-oven workers employed at the largest iron-steel factory in China were recruited for the study. Personal data were collected at the interview. DNA adduct levels in total white blood cells (WBCs) were detected using ³²P-postlabeling techniques. Genetic polymorphisms were analyzed by polymerase chain reaction (PCR) methods. Results: The subjects were divided into low and high exposure groups, according to personal exposure to PAHs. The mean adduct value was 1.57 (range 0.54 to 4.35) per 10⁸ nucleotides. A tendency for increased levels of DNA adducts in the high exposure group was observed, compared with the low exposure group (P = 0.07). In the low exposure group, DNA adducts were found to be positively associated with urinary cotinine (r = 0.44, P = 0.01). The rare allele homozygotes of CYP1A1 showed significantly higher DNA adduct levels than those of other CYP1A1 genotypes. Individuals with the NAT2 wild type had significantly increased DNA adduct levels than those with other NAT2 genotypes in the high exposure group. The p53 genetic polymorphism revealed a significantly positive effect on DNA adducts formation. There was a significantly higher adduct level in the subjects with a family history of cancer than those without, in the high exposure category. Conclusions: Effects of several variables, such as smoking, genetic polymorphism of 2 CYP1A1, NAT2, and gene p53, and a family history of cancer on DNA adduct levels were found, suggesting that these variables should be considered when evaluating the genotoxic effect of occupational exposure to PAHs using WBCs DNA adducts. Received: 15 February 1999 / Accepted: 15 June 1999     

Determinants of benzo[a]pyrene diol epoxide adducts to albumin in workers exposed to polycyclic aromatic hydrocarbons

The present study was undertaken among 260 subjects [133 controls and 127 persons exposed to polycyclic aromatic hydrocarbons (PAHs)] from two steel foundries and a graphite electrode producing plant in order to investigate the relationship between the benzo[a]-pyrene-diol epoxide adducts to albumin (BPDE-alb) and the intensity of exposure to PAHs. Blood samples were collected from each subject and BPDE-alb adduct determination was performed using a high-pressure liquid chromatography technique with fluorescence detection. Exposure to PAHs was assessed by measuring airborne concentration of 13 PAHs including benzo[a]pyrene (BaP) using personal air sampling and 1-hydroxypyrene excretion (1-HOP) in postshift urine. Significantly higher BPDE-alb adduct levels were observed in exposed workers compared to controls but wide interindividual variation was observed between subjects with the same level of exposure. BPDE-alb adduct level was weakly but significantly associated with the airborne concentrations of total PAHs (r = 0.35, P = 0.0001) and BaP (r = 0.30, P = 0.0001), and urinary 1-HOP excretion (r = 0.29, P = 0.0001). Alcohol and dietary habits, place of residence, and renal and hepatic status were not found to influence the concentration of BPDE-alb adducts significantly. However, for the same level of exposure to BaP, smokers had a higher probability of having an elevated BPDE-alb adduct level than nonsmokers. As long as BPDE-protein adduct levels have not been demonstrated to be better related to the risk of adverse genotoxic effects caused by PAHs in target organs than the 1-HOP level in urine, the latter remains the most practical biological parameter for assessing the risk of exposure to PAHs.     

Assessment of the bioavailability of PAHs in rats exposed to a polluted soil by natural routes: induction of EROD activity and DNA adducts and PAH burden in both liver and lung.

In order to assess bioavailability of polycyclic aromatic hydrocarbons (PAHs) present in soils, male laboratory rats were exposed to litters of control and polluted soils. After 88+/-2 h of exposure, several biomarkers were measured in both liver and lung. When rats were exposed to SIV soil, contaminated by a mixture of at least 13 PAHs, (1) only 2 or 3 PAH compounds were detected in liver and lung; (2) cytochrome P450-dependent monooxygenase activity, followed by 7-ethoxyresorufin O-deethylase (EROD) activity measurement, was highly induced in liver (13-fold-induction) and lung (up to 78-fold); and (3) DNA adducts were significantly increased. For what concerns soil artificially contaminated by only one PAH (phenanthrene or B[a]P), EROD activity was not or fully induced, respectively. These results demonstrate the occurrence of a high bioavailability of PAHs to mammals in natural conditions of exposure. First results concerning DNA adducts must be profound, but they already show that a short exposure of mammals to PAH-polluted soils can lead to potential genotoxic effects. EROD activity can be used as a sensitive biomarker in both liver and lung of rats maintained on litters of soils in the laboratory, and such a test can be used routinely to contribute to risk assessment.     

Using Semipermeable Membrane Devices, Bioassays, and Chemical Analysis for Evaluation of Bioavailable Polycyclic Aromatic Hydrocarbons in Water

Meiliang Bay is a sublake of Taihu Lake and has been polluted by domestic and industrial effluents. As part of a comprehensive risk assessment project in this region, semipermeable membrane devices (SPMDs) were applied to evaluate the levels and potential toxic potency of polycyclic aromatic hydrocarbons (PAHs) in lakewater, in combination with chemical analysis and in vitro bioassay using H4IIE rat hepatoma cells. In addition, induction of hepatic ethoxyresorufin-O-deethylase (EROD) activity, in crucian carp (Carassius auratus), caged in the vicinity of SPMD sampling sites was also used as a biomarker of exposure to PAHs and related chemicals. The caged crucian carp accumulated similar PAH profile patterns (p < 0.001) but at lower levels compared with SPMDs on a wet-weight basis. Total concentrations of PAHs in crucian carp muscle tissues ranged from 35.6 to 69.1 ng/g after the 32-day exposure, whereas total PAHs in SPMDs ranged from 716.9 to 1007.8 ng/g. Dialysates from SPMDs exposed to Meiliang Bay water caused marked EROD activity in H4IIE cells. Toxic potency of dialysates expressed as bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ) ranged from 3.8 to 6.2 pg TCDD-EQ/g SPMD for 32-day exposure samples. A linear correlation of total amount of PAHs and in vitro TCDD-EQ of SPMD dialysates yielded an R ² of 0.82. Empirical evidence suggests that aryl hydrocarbon receptor-active PAHs can account for about 42–56% of the potency observed. Significant induction of liver EROD activity was also observed in crucian carp caged alongside SPMDs and there was a good correlation between the results of EROD assays in vivo and in vitro (R ² = 0.62, p = 0.02). It is, therefore, suggested that the SPMD technique in connection with chemical analysis and specific in vitro bioassays might be a valuable tool to assess the levels and effects of bioavailable hydrophobic pollutants in water.     

Prenatal exposure to polycyclic aromatic hydrocarbons, benzo[a]pyrene-DNA adducts, and genomic DNA methylation in cord blood

Background: Polycyclic aromatic hydrocarbons (PAHs) are carcinogenic environmental pollutants generated during incomplete combustion. After exposure and during metabolism, PAHs can form reactive epoxides that can covalently bind to DNA. These PAH–DNA adducts are established markers of cancer risk. PAH exposure has been associated with epigenetic alterations, including genomic cytosine methylation. Both global hypomethylation and hypermethylation of specific genes have been associated with cancer and other diseases in humans. Experimental evidence suggests that PAH–DNA adduct formation may preferentially target methylated genomic regions. Early embryonic development may be a particularly susceptible period for PAH exposure, resulting in both increased PAH–DNA adducts and altered DNA methylation.Objective: We explored whether prenatal exposure to PAHs is associated with genomic DNA methylation in cord blood and whether methylation levels are associated with the presence of detectable PAH–DNA adducts.Methods: In a longitudinal cohort study of nonsmoking women in New York City, we measured PAH exposure during pregnancy using personal air monitors, assessed PAH internal dose using prenatal urinary metabolites (in a subset), and quantified benzo[a]pyrene–DNA adducts and genomic DNA methylation in cord blood DNA among 164 participants.Results: Prenatal PAH exposure was associated with lower global methylation in umbilical cord white blood cells (p = 0.05), but global methylation levels were positively associated with the presence of detectable adducts in cord blood (p = 0.01).Conclusions: These observations suggest that PAH exposure was adequate to alter global methylation in our study population. Additional epidemiologic studies that can measure site-specific cytosine methylation and adduct formation will improve our ability to understand this complex molecular pathway in vivo.     

Effect of Maintaining Rainbow Trout in Creosote Microcosms on Lens Optical Properties and Liver 7-Ethoxyresorufin-O-Deethylase (EROD) Activity

Previously, exposure of fish to polycyclic aromatic hydrocarbons (PAHs) in both field and laboratory settings has been associated with eye damage, but this has only been expressed qualitatively. In this study, an automated scanning laser system has been employed to quantitatively evaluate changes in lens optical quality in rainbow trout (Oncorhynchus mykiss) following their in vivo exposure to creosote, which is a complex mixture with many PAHs. Rainbow trout were placed in 12,000-L outdoor microcosms dosed with 0, 3, or 10 μl/L liquid creosote for a 28-day period. Collected fish were examined for changes in focal length variability (FLV), lens size, and weight. These measurements were compared with induction of hepatic ethoxyresorufin-O-deethylase (EROD) activity and hepatic and water concentrations of priority pollutant PAHs. The optical quality of rainbow trout lenses was significantly reduced following creosote exposure, as indicated by increased FLV. Lens damage was significantly related to hepatic EROD activity, and both effects rose with creosote dose. Analytical measurements of microcosm water indicated elevated concentrations of PAHs in creosote-dosed ponds, including compounds capable of inducing rainbow trout EROD activity in vitro. Hepatic concentrations of PAHs were low and not related to creosote dose, likely due to rapid metabolism and elimination. This study demonstrates for the first time employment of a highly sensitive and quantitative technique to measure lens damage in fish exposed to contaminants in situ. The relationship between this effect and hepatic CYP1A activity may suggest a mechanistic linkage, which could lead to the use of EROD activity as an indicator of toxic effect rather than just chemical exposure. Received: 17 May 1999/Accepted: 24 September 1999     

Aromatic DNA adducts in white blood cells of coke workers

Summary White blood cell DNA adducts were measured in coke workers, local controls and countryside controls using the ³²P-postlabelling technique. The method detected aromatic adducts including those formed by polycyclic aromatic hydrocarbons (PAHs). Coke workers are heavily exposed to PAHs particularly when working at the batteries. A difference in adduct levels was noted between the coke workers at the battery as compared to other jobs. The adduct levels in the non-battery were higher than those in the countryside controls.     

Polycyclic Aromatic Hydrocarbon Metabolites and 7-Ethoxyresorufin O-Deethylase Activity in Caged European Eels

This study investigated the contribution of two biomarkers, bile polycyclic aromatic hydrocarbon (PAH) metabolites and 7-ethoxyresorufin O-deethylase (EROD), activity in the assessment of PAH contaminated sites. European eels (Anguilla anguilla) were caged in a freshwater stream upstream and downstream from local industrial effluent outlets. Bile PAH metabolites were recorded as fluorescent aromatic compounds by synchronous fluorescence spectroscopy and as a marker for total PAH metabolism: 1-hydroxypyrene (1-OH Pyr) was isolated by high-pressure liquid chromatography and quantified. After 14 and 28 days of caging, EROD activity, bile fluorescence (synchronous fluorometric measurement), and 1-OH Pyr concentrations in bile were higher at the downstream site than at the upstream site. This increase was similar after 2 and 4 weeks of caging. During a reversibility study, EROD activity, bile fluorescence, and 1-OH Pyr concentrations decreased, and this trend was similar for the three markers. These results suggest that PAHs could be the main factor responsible for EROD induction in eels caged at the downstream site.     

Induction of EROD Activity and Genotoxic Effects by Polycyclic Aromatic Hydrocarbons and Resin Acids on the Juvenile Eel (Anguilla anguillaL.)

The purpose of this research is the study of total ethoxyresorufinO-deethylase (EROD) activity and erythrocytic nuclear abnormalities (NA) induction by polycyclic aromatic hydrocarbons (PAHs) and resin acids (RAs) on the juvenile eelAnguilla anguillaL. Juvenile eels were exposed, during 3 days, to the same molar concentration range (0, 0.1, 0.3, 0.9, and 2.7 μM) of β-naphthoflavone (BNF) and benzo[a]pyrene (BP)—PAHs—and abietic acid (AA) and dehydroabietic acid (DHAA)—RAs—and one alkylating agent, such as cyclophosphamide (CP). Results revealed that both PAHs are able to significantly increase total EROD activity at 0.9 and 2.7 μm, and both RAs at 0.3 μM, whereas NA frequency was significantly increased by BP at 0.9 μM and DHAA at 2.7 μM. BNF was found to be the most potent EROD inducer, and BP exhibited the highest mutagenic potential. An additional experiment was carried out in order to study the BNF total EROD and NA induction over a period of 3, 6, and 9 days. An increased length of exposure, beyond 3 days, does not proportionally increase total EROD activity. However, 6 and 9 days BNF exposure significantly increases NA, reaching the (NA) levels previously determined for 3 days 0.9 μM BP exposure.     

Concentrations of Selected Heavy Metals in Bones and Femoral Bone Structure of Bank (<Emphasis Type="Italic">Myodes glareolus</Emphasis>) and Common (<Emphasis Type="Italic">Microtus arvalis</Emphasis>) Voles from Different Polluted Biotopes in Slovakia

Concentrations of selected heavy metals in the femora and femoral bone structure of bank (Myodes glareolus) and common (Microtus arvalis) voles from different polluted biotopes in Slovakia (Kolíňany and Nováky sites) were investigated. Length, weight, and histological structure of vole bones were also analyzed. We observed higher concentrations of lead (Pb), iron (Fe), copper (Cu), and zinc (Zn) in the bones of both species from the Kolíňany site. Significant differences were observed in the concentration of Fe in bank and common voles (p < 0.05) and in the concentration of Zn (p < 0.05) in common voles. The animals from Nováky had higher concentrations of cadmium (Cd) and nickel (Ni) in their bones; however, the differences were not significant. The measured values for bone length and weight were higher in both species from Nováky (p < 0.05). We did not identify differences in qualitative histological characteristics of the femora between the voles (M. glareolus and M. arvalis separately) between the two biotopes. In addition, no statistically significant differences for any the measured variables of primary osteons’ vascular canals were observed. Correlation analysis in M. glareolus showed a strong positive relation between Cd and Ni (r = 0.52), Pb and bone weight (r = 0.53), Fe and bone weight (r = 0.52), and Fe and perimeter size of primary osteons’ vascular canals (r = 0.55). In common voles, a strong positive relation was found between Fe and Cu (r = 0.60) and between Fe and perimeter size of vascular canals of primary osteons (r = 0.55). Our results indicate that accumulation of some heavy metals is slightly increased in the femora of both species at Kolíňany.     

Ecotoxicity of Sediments Contaminated by the Oil Spill Associated with the Tanker “Prestige” Using Juveniles of the Fish Sparus aurata

In November 2002, the oil spill from the tanker Prestige in the Galician Coast caused an ecological catastrophe in Spain. The adverse effects associated with the contaminants bound to sediments were tested using juveniles of the fish Sparus aurata (seabream). The approach evaluates sediment quality by using an integrated assessment including chemical and ecotoxicological data. Sediment samples were physicochemically characterized, and the concentration of contaminants (polycyclic aromatic hydrocarbons—(PAHs) and metals) was measured. Different biomarkers of exposure (metallothioneins and ethoxyresorufin O-deethylase activity (EROD)) and biomarkers of effect (histopathology) were analyzed along the time. A multivariate analysis approach was used to correlate concentration of contaminants and sublethal effects measured in individuals of fish. Results show that increasing concentrations of PAHs in sediments were related to increased EROD activities and histopathological lesions. This is the first evidence showing adverse effects associated with petroleum contamination of PAHs in sediments after this spill, and it demonstrates the utility of the sublethal toxicity tests for monitoring the impact of petroleum spills.     

Evaluation of the Interactive Effects of Lead,Zinc and Benzo(k)fluoranthene on Crucian Carp,Carassius carassius,Using a Multiple Biomarker Approach

The aim of this study was to evaluate the interactive effects of the metals lead (Pb) and zinc (Zn) and the polycyclic aromatic hydrocarbon benzo(k)fluoranthene (BkF) on crucian carp (Carassius carassius). To this end, various biomarkers in the fish [7-ethoxyresorufin-O-deethylase (EROD); superoxide dismutase (SOD); malondialdehyde (MDA) and metallothioneins (MT)] were measured after 96 h of exposure. When compared to either treatment with Pb and Zn mixtures or BkF-only treatment, the treatment with BkF combined with Pb and Zn resulted in lower responses of EROD and MT synthesis activity, and higher SOD and MDA increases. These results indicated that the co-exposure of metals and polycyclic aromatic hydrocarbons may cause more severe oxidative stress on fish. However, the effect of these interactions on EROD and MT may lead to an underestimation of the actual exposure in the field and thereby should be carefully considered.     

Polycyclic aromatic hydrocarbon exposure in Steller's eiders (Polysticta stelleri) and Harlequin ducks (Histronicus histronicus) in the eastern Aleutian Islands, Alaska, USA

Seaducks may be affected by harmful levels of polycyclic aromatic hydrocarbons (PAHs) at seaports near the Arctic. As an indicator of exposure to PAHs, we measured hepatic enzyme 7-ethoxyresorufin-O-deethylase activity (EROD) to determine cytochrome P4501A induction in Steller's eiders (Polysticta stelleri) and Harlequin ducks (Histronicus histronicus) from Unalaska, Popof, and Unga Islands (AK, USA) in 2002 and 2003. We measured PAHs and organic contaminants in seaduck prey samples and polychlorinated biphenyl congeners in seaduck blood plasma to determine any relationship to EROD. Using Akaike's information criterion, species and site differences best explained EROD patterns: Activity was higher in Harlequin ducks than in Steller's eiders and higher at industrial than at nonindustrial sites. Site-specific concentrations of PAHs in blue mussels ([Mytilus trossilus] seaduck prey; PAH concentrations higher at Dutch Harbor, Unalaska, than at other sites) also was important in defining EROD patterns. Organochlorine compounds rarely were detected in prey samples. No relationship was found between polychlorinated biphenyl congeners in avian blood and EROD, which further supported inferences derived from Akaike's information criterion. Congeners were highest in seaducks from a nonindustrial or reference site, contrary to PAH patterns. To assist in interpreting the field study, 15 captive Steller's eiders were dosed with a PAH known to induce cytochrome P4501A. Dosed, captive Steller's eiders had definitive induction, but results indicated that wild Steller's eiders were exposed to PAHs or other inducing compounds at levels greater than those used in laboratory studies. Concentrations of PAHs in blue mussels at or near Dutch Harbor (approximately 1,180-5,980 ng/g) approached those found at highly contaminated sites (approximately 4,100-7,500 ng/g).     

Genotoxic risk assessment in white blood cells of occupationally exposed workers before and after alteration of the polycyclic aromatic hydrocarbon (PAH) profile in the production material: comparison with PAH air and urinary metabolite levels

Objective: Workers in various industries can be exposed to polycyclic aromatic hydrocarbons (PAHs). The relationship between biomarkers of genotoxic risk, PAH compounds in air (ambient monitoring) and PAH metabolites in urine (internal exposure) were studied in 17 workers exposed to PAHs in a fireproof-material producing plant before and 3 months after the PAH profile was altered in the binding pitch. Methods: Two biomarkers of exposure, specific DNA adducts of (±)-r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE) and non-specific DNA adduct of 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodGuo) were determined in white blood cells (WBCs). In addition, DNA strand breaks were analysed in lymphocytes by single-cell gel electrophoresis in a genotoxic risk assessment. Sixteen PAH compounds in air were determined by personal air sampling, and hydroxylated metabolites of phenanthrene, pyrene and naphthalene were determined in urine. Results: After substitution of the binding pitch the concentrations of benzo[a]pyrene in air decreased (P<0.01). No changes could be observed for pyrene, while levels of phenanthrene (P=0.0013) and naphthalene (P=0.0346) in air increased. Consequently, median DNA adduct rates of anti-BPDE decreased after alteration of the production material (from 0.9 to <0.5 adducts/10⁸ nucleotides). No changes in the excretion of 1-hydroxypyrene in urine could be determined, whereas increased levels of 1-, 2+9-, 3- and 4-hydroxyphenanthrene (P<0.0001) and 1-naphthol and 2-naphthol (P=0.0072) were found in urine. In addition, a statistically significant increase in DNA strand break frequencies (P<0.01) and elevated 8-oxodGuo adduct levels (P=0.7819, not statistically significant) were found in the WBCs of exposed workers 3 months after the PAH profile in the binding pitch had been altered. Conclusion: The results presented here show that the increased concentration of naphthalene and/or phenanthrene in the air at the work place could induce the formation of DNA strand breaks and alkali-labile sites in WBCs of exposed workers.     

Genotoxic and Hematological Effects in Children Exposed to a Chemical Mixture in a Petrochemical Area in Mexico

Children living in Coatzacoalcos, Veracruz, and in nearby surrounding areas are exposed to a mixture of pollutants from different sources. Previous studies in the area have reported genotoxic and haematotoxic compounds, such as lead (Pb), benzene, toluene, and polycyclic aromatic hydrocarbons (PAHs), in environmental and biological samples. The final toxic effects of these compounds are unknown because the toxic behaviour of each compound is modified when in a complex mixture. This is the first study on the exposure and effect of chemical mixtures on children who live near a petrochemical area. The aim of this study was to evaluate genotoxicity and haematological effects in children environmentally exposed to such mixtures and to determine whether the final effect was modified by the composition of the mixture composition. Biomarkers of exposure to Pb, benzene, toluene, and PAHs were quantified in urine and blood samples of 102 children. DNA damage was evaluated using comet assay, and haematological parameters were determined. Our results show that Pb and toluene did not surpass the exposure guidelines; the exposure was similar in all three localities (Allenede, Mundo Nuevo, and López Mateos). In contrast, exposure to PAHs was observed at three levels of exposure: low, medium, and high. The most severe effects of these mixtures were strictly related to coexposure to high levels of PAHs.     

DNA adducts, benzo(a)pyrene monooxygenase activity, and lysosomal membrane stability in Mytilus galloprovincialis from different areas in Taranto coastal waters (Italy)

The aim of this study was to investigate the impact of environmental pollution at different stations along the Taranto coastline (Ionian Sea, Puglia, Italy) using several biomarkers of exposure and the effect on mussels, Mytilus galloprovincialis, collected in October 2001 and October 2002. Five sampling sites were compared with a "cleaner" reference site in the Aeronautics Area. In this study we also investigated the differences between adduct levels in gills and digestive gland. This Taranto area is the most significant industrial settlement on the Ionian Sea known to be contaminated by polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls, heavy metals, etc. Exposure to PAHs was evaluated by measuring DNA adduct levels and benzo(a)pyrene monooxygenase activity (B(a)PMO); DNA adducts were analyzed by 32P-postlabeling with nuclease P1 enhancement in both gills and digestive glands to evaluate differences between DNA adduct levels in the two tissues. B(a)PMO was assayed in the microsomal fraction of the digestive glands as a result of the high expression of P450-metabolizing enzymes in this tissue. Lysosomal membrane stability, a potential biomarker of anthropogenic stress, was also evaluated in the digestive glands of mussels, by measuring the latent activity of beta-N-acetylhexosaminidase. Induction of DNA adducts was evident in both tissues, although the results revealed large tissue differences in DNA adduct formation. In fact, gills showed higher DNA adduct levels than did digestive gland. No significant differences were found in DNA adduct levels over time, with both tissues providing similar results in both years. DNA adduct levels were correlated with B(a)PMO activity in digestive gland in both years (r = 0.60 in 2001; r = 0.73 in 2002). Increases were observed in B(a)PMO activity and DNA adduct levels at different stations; no statistical difference was observed in B(a)PMO activity over the two monitoring campaigns. The membrane labilization period in mussels from some stations was decreased in both years. No statistical differences were established in the membrane labilization times from 2001 to 2002. Our results suggest the existence of different sources and amounts of environmental contaminants at the stations investigated. The formation of DNA adducts confirms the existence of activation pathways in mussels and shows the importance of DNA adduct analysis in the gill tissue in addition to the more commonly used digestive gland; these results confirm the utility of lysosomal membrane stability as a biomarker of general stress. Overall, the integrated use of biomarkers of exposure and the effects of environmental contaminants on living marine organisms may help to better interpret the impact of pollutants in a marine coastal environment.     

The Dose–Response Relationships for EROD and GST Induced by Polyaromatic Hydrocarbons in <Emphasis Type="Italic">Carassius auratus</Emphasis>

Freshwater fish Carassius auratus were chosen as experimental animals, the hepatic biochemical responses to medium-term exposure of five PAHs were measured as ethoxyresorufin O-deethylase (EROD) activity (phase I) and glutathione S-transferase (GST) activity (phase II) to assess sub-lethal effects. The fold increases of EROD and GST activity were calculated and both increased in the order Fluoranthene < Fluorene < Benzo(b)fluoranthene < Benzo(g,h,i)perylene < Indeno(1,2,3-cd)-pyrene. The clear dose–response relationships were found for liver EROD and GST activity induced by PAHs. The enzyme EROD and GST in Carassius auratus were confirmed as useful biomarkers of exposure to both PAH and PAH-like compounds.