Tissue distribution and temperature-dependence of Anguilla anguilla L. EROD activity following exposure to model inducers and relationship with plasma cortisol, lactate and glucose levels

Anguilla anguilla L. ethoxyresorufin-O-deethylase (EROD) elevation by 2.7 microM beta-naphthoflavone (BNF) 3 days water exposure, or 4 mg/kg ip exposure, was studied in four different organs--liver, kidney, gills, and intestine. The results demonstrated a significant increase in liver EROD activity for the two previous conditions, whereas kidney EROD activity only increased during the intraperitoneal exposure. A. anguilla was also exposed during 8, 16, and 24 h to water contaminated with 2.7 microM BNF or benzo[a]pyrene (BaP) at 20 degrees C and 25 degrees C. Both compounds significantly increased liver EROD activity from 8 up to 24 h. There was no significant difference in liver EROD activity elevation by both compounds, either at 20 degrees C or 25 degrees C. Liver EROD activity was demonstrated to be one of the first warning systems concerning the presence of polycyclic aromatic hydrocarbons (PAHs) in water. A. anguilla 3 h exposure to diesel oil water-soluble fraction (DWSF) significantly increased plasma cortisol and significantly decreased plasma lactate. A prolonged exposure beyond 3 h, i.e. 4 h, 2, 3, 4, and 6 days to the previous conditions demonstrated a significant liver EROD activity elevation from Day 2 up to 6, and a significant increase in erythrocytic nuclear abnormalities (ENA) at Day 6.     

Induction of EROD activity in European eel (Anguilla anguilla) experimentally exposed to benzo[a]pyrene and beta-naphthoflavone

The induction of liver ethoxyresorufin O-deethylase (EROD) activity was investigated in the European eel, Anguilla anguilla, collected from a Mediterranean brackish environment and experimentally exposed to benzo[a]pyrene (B[a]P) and beta-naphthoflavone (BNF). Eels were injected intraperitoneally at increasing doses (0.1, 1, 10, and 50 mg/kg wet body weight) using corn oil as a carrier and sacrificed after 7 days. The main objectives of the present study are: (1). to assess of the sensitivity of EROD induction as a biomarker to polycyclic aromatic hydrocarbon (PAH) exposure; (2). to determine an EROD dose-response relationship of the contaminants used; and (3). to compare the efficiency of B[a]P and BNF as inducers of EROD activity. Results showed that both chemicals resulted in a dose-dependent EROD induction, but increases were not linear. EROD activity seemed to reach a plateau at the exposure of 10 mg/kg in both treatment groups; B[a]P was a more potent inducer than BNF was at the higher doses (10 and 50 mg/kg), while the opposite result was observed at the lower ones (0.1 and 1 mg/kg). The greatest induction occurred in eels treated with 10 mg/kg B[a]P, in which a 261-fold increase in EROD activity was observed. Results showed that EROD activity in A. anguilla is significantly induced by B[a]P and BNF exposure, responding to a wide range of concentrations of these contaminants. We infer that this tool may be suited as a diagnostic biomarker for biomonitoring PAHs pollution in Mediterranean brackish environments and further field research is suggested.     

Endocrine and metabolic changes in Anguilla anguilla L. following exposure to beta-naphthoflavone--a microsomal enzyme inducer

Anguilla anguilla L. were exposed during 24 and 48 h to 2.7 muM beta-naphthoflavone (BNF), a known microsomal enzyme inducer. The BNF effects on thyroid-stimulating hormone (TSH), free triiodothyronine (T3), free thyroxine (T4) and cortisol plasma levels were investigated. Alterations on plasma glucose and lactate levels were also measured as an indication of energy-mobilizing hormones alterations. BNF showed to be able to decrease significantly A. anguilla plasma T4 levels, whereas TSH, T3 and cortisol plasma remained constant. However, plasma glucose levels were significantly increased, demonstrating that intermediary metabolism has been affected. These results demonstrate that BNF a PAH-like compound alters the normal functioning of the hypothalamo-pituitary-thyroid (HPT) axis in A. anguilla.     

Genotoxic and biochemical responses in caged eel (Anguilla anguilla L.) after short-term exposure to harbour waters

European eel (Anguilla anguilla L.) were caged and exposed in situ for 8 and 48 h to the Aveiro offward fishing harbour water (HW) and to clean seawater under laboratory conditions (Control). Eel liver biotransformation (Phase I) was measured as ethoxyresorufin-O-deethylase (EROD) activity, cytochrome P450 (P450) and glutathione S-transferase (GST) activity (Phase II). Genotoxic responses were determined as blood, liver and kidney DNA strand breaks as well as erythrocytic nuclear abnormalities (ENAs). HW failed to significant increase liver EROD, GST activities and ENA frequency. Nevertheless, P450 content was significantly increased after 8 and 48 h exposure. Genotoxicity measured as DNA integrity decrease was found in blood after 8 and 48 h exposure to HW, whereas in liver and kidney, it was observed after 48 h exposure to HW. Blood, kidney and liver genotoxicity may be due to the presence of polycyclic aromatic hydrocarbons (PAHs) which are genotoxic compounds and the main HW organic contaminants.     

Anguilla anguilla L. plasma cortisol, lactate and glucose responses to abietic acid, dehydroabietic acid and retene

Anguilla anguilla L. were exposed to 0, 0.1, 0.3, 0.9 and 2.7 microM abietic (AA), dehydroabietic (DHAA) acids and retene (Re) during 8, 16, 24 and 72 h. The eels plasma cortisol, glucose and lactate were measured. A significant decrease in plasma cortisol was observed at 72 h exposure to 0.9 and 2.7 microM Re. DHAA (0.1 microM) significantly decreased plasma cortisol in eels after 8 and 24 h exposure. However, a significant plasma cortisol increase was found after 16 h, 2.7 microM AA exposure and after 24 h exposure to 0.1 microM and 2.7 microM AA. Furthermore, 72 h exposure to 0.9 microM AA also induced a plasma cortisol increase. A general rise in plasma glucose was detected after all exposure periods to Re. The plasma lactate also increased after 72 h exposure to 2.7 microM AA and after 8 h exposure to 0.1 microM DHAA.     

Biotransformation, stress and genotoxic effects of 17beta-estradiol in juvenile sea bass (Dicentrarchus labrax L.)

The effects of 17beta-estradiol (E(2)) on fish became a matter of concern, since significant levels of this hormone were detected in the aquatic environment released mainly by domestic sewage treatment plants. In this perspective, the current study was focused on E(2) effects upon biotransformation, stress and genotoxic responses of juvenile Dicentrarchus labrax L. (sea bass). Fish were exposed to E(2) during 10 days in two different ways: water diluted (200 ng/L or 2,000 ng/L) and i.p. injected (0.5 mg/kg or 5 mg/kg). A battery of biological responses was evaluated: liver ethoxyresorufin-O-deethylase (EROD) and alanine transaminase (ALT) activities, liver somatic index (LSI), plasma cortisol, glucose and lactate concentrations, as well as erythrocytic nuclear abnormalities (ENA). All the exposure conditions induced endocrine disruption, measured as plasma cortisol decrease, and genotoxicity, measured as ENA increase. Thus, no differences were detected either between different exposure routes or tested concentrations. Concerning liver EROD and ALT activities, as well as plasma glucose and lactate concentrations no differences were found between treated and control groups. LSI was the only parameter to respond differently in the two exposure routes, as only E(2) water diluted induced a significant increase in this hepatic indicator.     

Genotoxic effects of binary mixtures of xenoandrogens (tributyltin, triphenyltin) and a xenoestrogen (ethinylestradiol) in a partial life-cycle test with Zebrafish (Danio rerio)

A partial life-cycle test with the model fish Danio rerio was performed in order to evaluate the genotoxic potential of binary mixtures of xenoandrogenic (tributyltin--TBT; triphenyltin--TPT) and an estrogenic compound (ethinylestradiol--EE2). Five days post-fertilisation larvae were diet-exposed to environmental relevant concentrations of TBT and TPT (25 ng/g-100 ng/g), and water-exposed to ethinylestradiol (3.5 ng/L) for a four-month period; binary mixtures of TBT plus EE2 and TPT plus EE2 were run in parallel. The erythrocytic nuclear abnormalities (ENA) assay in circulating erythrocytes was used to evaluate genotoxicity in the end of the four-month exposure period. A significant increase (p<0.05, Kruskall-Wallis non-parametric ANOVA) in ENA frequency, in comparison with control animals, was observed in those animals exposed to TBT and TPT (the highest doses only), and to EE2 and the binary mixtures, although neither synergistic nor additive effects of the tested compounds were evident. Overall, the results clearly indicate that chronic exposure to low levels of TBT, TPT, EE2 and binary mixtures of TBT plus EE2 and TPT plus EE2 are genotoxic to zebrafish, which may suggest that wild fish populations may be under increased DNA damage in areas contaminated by these endocrine disrupting chemicals.     

Toxic effects of waterborne polychlorinated biphenyls and sex differences in an endangered goodeid fish (Girardinichthys viviparus)

Polychlorinated biphenyls (PCBs) elicit toxic effects in different species. PCBs undergo biotransformation by enzymes associated with the mixed functional oxidase system, such as cytochrome P450 (cyt P450), this biotransformation being sex-dependent. No other metabolic pathways are known, however, in connection with this process. Thus, the present study aims to evaluate the effects of waterborne PCBs at sublethal nominal concentrations (0.92 mg PCBs/L) on the black-fin goodeid Girardinichthys viviparus, an endangered fish native to the Valley of Mexico, as well as any sex differences related to PCB biotransformation. Eight-month-old adult fish born in the laboratory were exposed for 1, 2, 4, 8 and 16 days to half the LC(0) (calculated concentration at which no deaths occurred after 96 h) determined by an acute toxicity test. Specimens were sacrificed following exposure and the liver was used to evaluate different biomarkers: cyt P450 concentration and alcohol dehydrogenase (ADH) and ethoxyresorufin O-deethylase (EROD) activities. Results show sexual differentiation with regard to all biomarkers in both the control group and PCB-treated fish, with higher values found in males. The induction rate of cyt P450 remained constant throughout the study in males. In females, induction peaked on day 4, coinciding with maximum EROD activity, and fell significantly thereafter. EROD was lower in PCB-treated males than in the control group, but was greater in magnitude. ADH was significantly induced in both sexes from day 2 to day 16 of exposure. The highest response as compared to the control group occurred on day 8 in females. A correlation was found between ADH activity and exposure to PCBs. Three possible action mechanisms, operating either individually or concurrently, are proposed.     

Anguilla anguilla L. antioxidants responses to in situ bleached kraft pulp mill effluent outlet exposure

This study assesses the antioxidant enzymes activities viz., catalase, glutathione peroxidase, glutathione S-transferase and nonenzymatic antioxidant molecule such as glutathione in Anguilla anguilla L. gill, kidney and liver in response to 8- and 48-h exposure to bleached kraft pulp mill effluent (BKPME). A. anguilla were caged and plunged at three different sites-50 (Site 1), 100 (Site 2) and 2000 m (Site 3) away from the closed BKPME outlet. A significant gill (8 and 48 h) and kidney (48 h) catalase activity decrease was observed at site 2 exposure whereas liver showed a significant increase in catalase activity after 8 and 48 h to site 1 exposure. Glutathione peroxidase (GPX) activity was significantly decreased in gill after 8-h exposure to site 1 and 48-h exposures to sites 1 and 2, respectively. Concerning gill, kidney and liver glutathione S-transferase (GST) activity, a significant gill GST activity decrease after 8 h at site 2 and 48 h at sites 1 and 2 was observed; in kidney, a significant decrease in its activity was observed after 48 h at sites 1 and 2, respectively, whereas in liver, the decrease was significant only at site 2 after 48-h exposure. The in situ BKPME exposure caused a significant total gill and kidney reduced glutathione (GSH) decrease after 8 h at site 2 exposure and after 48 h at site 1 and 2 exposures, respectively. However, a biphasic response was observed in liver, i.e. initial significant increase after 8 h at site 2 followed by a significant decrease after 48 h to the same site exposure. The enzymatic and nonenzymatic antioxidants pattern in gill and kidney, as observed in this study, was different than liver, demonstrating that the liver was more resistant to oxidative damage than gill and kidney. In addition, A. anguilla gill, kidney and liver antioxidants adaptation potentials may serve as a surrogate biomarker to BKPME exposure.     

Contrasting retinoid and thyroid hormone status in differentially-contaminated northern fulmar colonies from the Canadian Arctic,Svalbard and the Faroe Islands

The northern fulmar (Fulmarus glacialis) has previously been shown to accumulate a wide range, and occasionally high concentrations of organochlorines (OCs) (e.g., PCBs, chlorobenzenes, DDT- and chlordane-related compounds, dioxins and furans). The present study aimed to investigate, using a meta-analysis approach, the variations in cytochrome P450 (CYP) 1A-like enzyme induction based on ethoxyresorufin O-deethylase activity (EROD) and selected physiological variables (retinoids and thyroid hormones) in northern fulmar breeding in three differentially OC-exposed populations: Nunavut (Canadian Arctic), Svalbard (Norwegian Arctic) and the Faroe Islands. Substantially higher (roughly two-fold) OC levels were uncovered in the liver of this long-lived fulmarine petrel breeding in the Faroe Islands relative to Svalbard and Nunavut. Liver levels of PCDDs, PCDFs and non-ortho PCBs in Faroe Islands fulmars were amongst the highest reported thus far in any seabirds from the northern regions. Positive correlations were depicted in combined fulmars (all three populations) between hepatic EROD activity and concentrations of OCs, in which strongest associations were found for dioxin-like compound (PCDFs and PCDDs) and TEQ concentrations. Moreover, moderate to strong positive correlations were found between liver OC concentrations and plasma total thyroxin (TT₄) levels and TT₄/total triiodothyronine (TT₃) level ratios, as well as strong negative correlations between the same suite of OCs and plasma TT₃ levels. Hepatic OC concentrations (PCBs, PCDDs, PCDFs, HCB, p,p'-DDE and oxychlordane) also were positively correlated with hepatic retinyl palmitate levels which, in turn, were associated with a significant decrease in plasma retinol levels and somewhat unchanged liver retinol levels. The present meta-analysis investigation on northern fulmar breeding in three geographically-distant sites illustrated that OC exposure (mainly PCBs and dioxins/furans) may be associated with modulation of the thyroid and retinoid homeostasis. However, the impact of confounding environmental factors (e.g., temperature and nutritional status) on current physiological variable variations could not be ruled out, and thus any cause-effect linkages between thyroid and retinoid system perturbation and OC exposure cannot be ascertained.     

The mutagenic activity of thirty polycyclic aromatic hydrocarbons (PAH) and oxides in urban airborne particulates

Data are presented on the mutagenicity of an organic extract of a composite sample of urban air particulates and of thirty PAH compounds in such samples, including four quinone derivatives, isolated quantitatively by thin-layer chromatography and identified by fluorescence or other spectral techniques. Mutagenic activity was determined by the Ames assay, using histidine auxotrophs of Salmonella typhimurium strains TA98, TA100, TA1537 and TA1538. All compounds were dissolved in dimethyl sulfoxide (DMSO) which was the least toxic of eight organic solvents tested.The mutagenic activity of a benzene extract of suspended particulates from the air of Hamilton, Ontario was significantly greater with strain TA98 than with strain TA100, suggesting the presence of more frame-shift acting mutagens. The mutagenic response of this extract was similar with and without S-9 activation.Mutagenic tests on the 30 PAH compounds indicated that only the benzo(a)pyrene quinones were direct acting mutagens. All of the chemical compounds were tested with and without S-9 activation. The following PAH showed unequivocal mutagenic activity, with S-9 activation: benz(a)anthracene, benzo-(a)pyrene, benzo(ghi)perylene, benzo(rst)pentaphene, benzophenanthrene, chrysene, 1, 2, 3, 4-dibenzanthracene, 2, 3, 6, 7-dibenzanthracene, and 3-methyl cholanthrene. The quinones of 1, 6-; 3, 6-; and 6, 12-benzo(a)pyrenes showed weak mutagenic activity but 3, 6-benzo(a)pyrene elicited also a photodynamic response.     

A bioassay approach to determine the dioxin-like activity in sediment extracts from the Danube River: ethoxyresorufin-O-deethylase induction in gill filaments and liver of three-spined sticklebacks (Gasterosteus aculeatus L.)

Sediment samples from the upper Danube River in Germany have previously been characterized as ecotoxicologically hazardous and contaminants in these sediments may contribute to the observed decline of fish populations in this river section. For the investigation of sediment toxicity there is a need for development, standardization and implementation of in vivo test systems using vertebrates. Therefore, the main objective of this study was to apply and evaluate a recently established fish gill EROD assay as a biomarker in sediment toxicity assessment by using extracts of well characterised sediment samples from the upper Danube River. This to our knowledge is the first application of this novel assay to sediment extracts. Sediments from four different sites along the upper Danube River were Soxhlet-extracted with acetone and dissolved in DMSO. Three-spined sticklebacks (Gasterosteus aculeatus L.) were exposed for 48 h to various concentrations of the extracts, to the positive control beta-naphthoflavone or to the solvent. Measurements of EROD activity in gill filaments and liver microsomes followed the exposure. Concentration-dependent induction of EROD in both gill and liver was found for all sediment extracts. The highest EROD-inducing potency was determined for extracts of sediments from the sites "Opfinger See" and "Sigmaringen" and the EROD activities in gill and liver correlated well. The results from the gill and liver assays were in accordance with in vitro results of previous investigations. The EROD activities measured in the present study corresponded with the concentrations of PAHs, PCBs and PCDD/Fs in the sediment samples derived in a previous study. The sticklebacks in this study were in the reproductive phase and a stronger EROD induction was obtained in the females than in the males. Implementation of the EROD assay in testing of sediment extracts gave highly reliable results which make this assay an ecotoxicologically relevant method for assessment of contamination with Ah receptor agonists in sediments.     

A review of airborne polycyclic aromatic hydrocarbons (PAHs) and their human health effects

Polycyclic aromatic hydrocarbons (PAHs) are a large group of organic compounds comprised of two or more fused benzene rings arranged in various configurations. PAHs are widespread environmental contaminants formed as a result of incomplete combustion of organic materials such as fossil fuels. The occurrence of PAHs in ambient air is an increasing concern because of their carcinogenicity and mutagenicity. Although emissions and allowable concentrations of PAHs in air are now regulated, the health risk posed by PAH exposure suggests a continuing need for their control through air quality management. In light of the environmental significance of PAH exposure, this review offers an overview of PAH properties, fates, transformations, human exposure, and health effects (acute and chronic) associated with their emission to the atmosphere. Biomarkers of PAH exposure and their significance are also discussed.     

Anguilla anguilla L. Genotoxic responses after in situ exposure to freshwater wetland (Pateira de Fermentelos, Portugal)

Pateira de Fermentelos is a Cértima River enlargement, close to its river mouth (by the Agueda River), where the introduction of agricultural chemicals such as fertilisers and pesticides, domestic sewage, as well as heavy metals from electroplating industries, results in increased water pollution. The present research work concerns a 48 h in situ exposure of caged eels (Anguilla anguilla L.) at the Pateira de Fermentelos. Five exposure sites were selected, i.e., site A, site B, site C, site D and site E in order to study its water genotoxicity potential, measured in gill, blood, liver and kidney as DNA strand breaks. Eels were also exposed at a reference site by the Cértima River spring. Bottom water samples were collected for further physical-chemical analysis. Site A exposure, significantly decreased gill, blood and liver DNA integrity. Gill and liver DNA integrity was also significantly decreased at site B. At site C only blood DNA integrity was significantly decreased. The present field in situ study demonstrated that the three exposure sites close to the Pateira initial part, such as A, B and C are polluted by pro and/or genotoxic compounds. The genotoxic effects induced in A. anguilla L. suggest a different contamination of the exposure sites A, B and C, in genotoxic chemicals. Thus, according to its genotoxic potential the exposure sites A, B and C, may be ordered as follows site A>site B>site C. No genotoxic effects on A. anguilla L. were observed at site E as DNA strand breaks increase.     

Spatiotemporal analysis and human exposure assessment on polycyclic aromatic hydrocarbons in indoor air,settled house dust,and diet: A review

This review summarizes the published literature on the presence of polycyclic aromatic hydrocarbons (PAH) in indoor air, settled house dust, and food, and highlights geographical and temporal trends in indoor PAH contamination. In both indoor air and dust, ΣPAH concentrations in North America have decreased over the past 30 years with a halving time of 6.7 ± 1.9 years in indoor air and 5.0 ± 2.3 years in indoor dust. In contrast, indoor PAH concentrations in Asia have remained steady. Concentrations of ΣPAH in indoor air are significantly (p < 0.01) higher in Asia than North America. In studies recording both vapor and particulate phases, the global average concentration in indoor air of ΣPAH excluding naphthalene is between 7 and 14,300 ng/m³. Over a similar period, the average ΣPAH concentration in house dust ranges between 127 to 115,817 ng/g. Indoor/outdoor ratios of atmospheric concentrations of ΣPAH have declined globally with a half-life of 6.3 ± 2.3 years. While indoor/outdoor ratios for benzo[a]pyrene toxicity equivalents (BaP_eq) declined in North America with a half-life of 12.2 ± 3.2 years, no significant decline was observed when data from all regions were considered. Comparison of the global database, revealed that I/O ratios for ΣPAH (average = 4.3 ± 1.3), exceeded significantly those of BaP_eq (average = 1.7 ± 0.4) in the same samples. The significant decline in global I/O ratios suggests that indoor sources of PAH have been controlled more effectively than outdoor sources. Moreover, the significantly higher I/O ratios for ΣPAH compared to BaP_eq, imply that indoor sources of PAH emit proportionally more of the less carcinogenic PAH than outdoor sources. Dietary exposure to PAH ranges from 137 to 55,000 ng/day. Definitive spatiotemporal trends in dietary exposure were precluded due to relatively small number of relevant studies. However, although reported in only one study, PAH concentrations in Chinese diets exceeded those in diet from other parts of the world, a pattern consistent with the spatial trends observed for concentrations of PAH in indoor air. Evaluation of human exposure to ΣPAH via inhalation, dust and diet ingestion, suggests that while intake via diet and inhalation exceeds that via dust ingestion; all three pathways contribute and merit continued assessment.     

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.     

Seasonal assessment of environmental tobacco smoke and respirable suspended particle exposures for nonsmokers in Bremen using personal monitoring

The study was designed to determine seasonal differences in personal exposures to respirable suspended particles (RSP) and environmental tobacco smoke (ETS) for nonsmokers in Bremen, Germany. The subjects were office workers, either living and working in smoking locations or living and working in nonsmoking locations. One hundred and twenty four randomly selected nonsmoking subjects collected air samples close to their breathing zone by wearing personal monitors for 24 h or, in some cases, for 7-day periods during the winter of 1999. The investigation was repeated in the summer with 126 subjects, comprised of as many of the studied winter population (89 subjects) as possible. Saliva cotinine analyses were undertaken to verify the nonsmoking status of the subjects. Subjects wore one personal monitor while at work and one while away from the workplace on weekdays, and a third monitor at the weekend. Collected air samples were analysed for RSP, nicotine, 3-ethenylpyridine (3-EP) and ETS particles. The latter were estimated using ultraviolet absorbance (UVPM), fluorescence (FPM) and solanesol (SolPM) measurements. ETS exposure was consistently higher in the winter than in the summer, this pattern being particularly evident for subjects both living and working with smokers. The highest median 24-h time weighted average (TWA) concentrations of ETS particles (SolPM, 25 microg m(-3)) and nicotine (1.3 microg m(-3)) were recorded for subjects performing weekday monitoring during the winter. These were significantly higher than equivalent levels of ETS particles (SolPM, 2.4 microg m(-3)) and nicotine (0.26 microg m(-3)) determined during the summer. There were no appreciable differences between winter and summer percent workplace contributions to median TWA ETS particle and nicotine weekday concentrations, the workplace in Bremen, in general, contributing between 35% and 61% of reported median concentrations. Workers, on average, spent one-third of their time at work during a weekday, indicating that concentrations were either comparable or higher in the workplace than in the home and other locations outside the workplace. Median 24-h weekend ETS particle and nicotine concentrations for smoking locations were not significantly different from equivalent weekday levels during the winter, but were significantly lower during the summer. Based upon median 24-h TWA SolPM and nicotine concentrations for the winter, extrapolated to 1 year's ETS exposure, those subjects both living and working in smoking locations (the most highly exposed group) would potentially inhale 13 cigarette equivalents/year (CEs/y). However, based on a similar extrapolation of summer measurements, the same group of subjects would potentially inhale between 1.3 and 1.9 CEs/y. The most highly exposed subjects in this study, based upon 90th percentile concentrations for those both living and working in smoking locations during the winter, would potentially inhale up to 67 CEs/y in the winter and up to 22 CEs/y in the summer. This clearly demonstrates that seasonal effects should be taken into account in the design and interpretation of ETS exposure studies. Air sampling over a 7-day period was shown to be technically feasible, and subsequent RSP, ETS particle and nicotine levels determined by 7-day monitoring were not found to be significantly different from equivalent levels determined by 24-h monitoring. However, the longer sampling period resulted in the collection of an increased quantity of analytes, which improved the limits of quantitation (LOQ) and allowed a more accurate determination of low level ETS exposure. This was reflected by a reduced percentage of data falling below the LOQ for 7-day monitoring compared with 24-h monitoring. The use of a liquid chromatographic method with tandem mass spectrometric detection for saliva cotinine measurement afforded a greatly improved LOQ and greater accuracy at low concentrations compared with the radioimmunoassay (RIA) method used in previous studies by these authors. In this study, 17 subjects out of 180 tested (9.4%) were found to have saliva cotinine levels exceeding the selected threshold of 25 ng ml(-1) used to discriminate between smokers and nonsmokers.     

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.     

The effect of iron on the biological activities of erionite and mordenite

Epidemiological data has demonstrated that environmental and/or occupational exposure to mineral particulates may result in the development of pulmonary fibrosis, bronchogenic carcinoma and malignant mesothelioma many years following exposure. It has been suggested that the genotoxic effects of fibrous particulates, such as asbestos, is due in part to the generation of reactive oxygen species (ROS) from iron associated with the particulates. However, the molecular mechanisms by which mineral particulates induce ROS that results in genotoxic damage remains unclear. The naturally occurring zeolites, erionite and mordenite share several physiochemical properties but they elicit very different biological responses, with erionite, a fibrous particulate, being highly toxic, and mordenite, a nonfibrous particulate, being relatively benign. We are using these natural zeolites as a model system to determine what physicochemical properties of these zeolites are responsible for their biological response(s) and to evaluate the parameters that influence these responses. The purpose of the present study was to determine the mutagenic potential of erionite and mordenite and to determine whether this mutagenic potential was modulated by iron. The results of this study using the Chinese hamster ovary cell line AS52 demonstrated that erionite was more cytotoxic than mordenite. However, the cytotoxicity of both zeolites was increased in the presence of physiological concentrations of ferrous chloride. Ferrous ions (5-20 microM) significantly (p<0.001) increased the cytotoxicity of mordenite, but only at the highest concentration (16 microg/cm(2)) of mordenite tested. Conversely, only the highest concentration (20 microM) of ferrous ion significantly (p<0.001) increased the cytotoxicity of erionite, but this enhanced cytotoxicity occurred over a wider concentration range (6-16 microg/cm(2)) of erionite. Mordenite was not mutagenic at any of the concentrations tested, and the mutagenic potential of mordenite was not enhanced by the addition of ferrous ion. Conversely, erionite was mutagenic in a dose-response manner at concentrations greater than 6 microg/cm(2) and the mutagenic potential of erionite was significantly enhanced by the addition of ferrous ions. These results suggest that while the cytotoxicity of mordenite and erionite may be related to the ability of these fibers to transport iron into a cell, the different coordination state of iron associated with the two fiber surfaces is critical for inducing genotoxic damage.     

Body burden of contaminants and biological effects in mussels: an integrated approach

An investigation of contaminants and biological effects in mussels from Halifax Harbour, Nova Scotia, Canada, focused on a 6 km section in the central most industrialized core of the harbour, where a site was previously identified as highly contaminated. The aim of the study was to compare the body burden of mussels in terms of polycyclic aromatic hydrocarbons (PAH), dichlorophenyltrichloroethane (DDT family), coprostanol and elements, relative to biological parameters such as condition indices, sex ratio, survival time in air, and to biochemical indicators of mussels' health analysed in gills, digestive gland and gonad tissues. These markers are total sugar and lipid content of gonads, mitochondrial electron transport activity in digestive gland and gonad tissues, lipid peroxidation in gill, digestive gland and gonad tissues, and heme oxidase activity in the digestive gland. At the north western end of the area, near a major sewage effluent, shorter survival time, higher oxidative stress and metabolism, gonad electron transport activity, levels of coporostanol, PAH, p,p'-dichlorophenyldichlo-roethylene (p,p'-DDE), Ag, Cu, Fe and P were observed. At the opposite south eastern end, longer survival time, higher lipid content, lowest condition indices and concentrations of coprostanol, PAH and p,p'-DDE, but higher concentrations of Sn and Cd, were detected. On-going improvements to sewage treatment in Halifax Harbour, including construction of sewage treatment plants that will discharge into deeper parts of the central harbour, should improve inter-tidal mussels' health in our study area.