Identification of cis- and trans-verbenol in human urine after occupational exposure to terpenes

Summary Urine from sawmill workers exposed to -pinene, -pinene and -3-carene was collected and hydrolyzed with -glucuronidase at pH 5.0 for 24h at 37°C. After hydrolysis the urine was cleaned on a SEP-PAK C₁₈ cartridge. The cartridge was eluted with n-heptane. The eluate was injected onto a gas chromatograph equipped with a 25-m (0.32-mm ID) SP-1000 capillary column. The major peak in the chromatogram was identified by GC-MS as trans-verbenol by electron impact at 70 eV. cis-Verbenol was also identified. These metabolites could not be detected in non-hydrolyzed urine from the exposed workers or in hydrolyzed urine from an unexposed individual. The recoveries of the verbenols from hydrolyzed urine were in the range of 85 to 94% and the metabolites were stable both in urine and in n-heptane after sample cleaning at –20°C for at least 12 weeks. We suggest that these metabolites are formed from -pinene by hydroxylation.     

The use of respiratory enzymes as biomarkers of petroleum hydrocarbon exposure in Mytilus edulis planulatus

The effect of exposure to petroleum hydrocarbons via the water column and through contaminated sediment upon changes in respiratory enzymes in the common mussel (Mytilus edulis planulatus) was investigated. Mussels were exposed to three concentrations of the water-accommodated fraction (WAF) of Bass Strait crude oil, for 24, 48, and 96 h. In a second study mussels were exposed to three concentrations of crude oil-contaminated sediment for 2 weeks and 1, 2, 4 and 6 months. Activities of citrate synthase (CS) and lactate dehydrogenase (LDH) were measured in the gills. In mussels exposed to WAF, a significant decrease in CS activity was observed over time (P<0.05), whereas treatment did not cause a significant change in CS activity (P>0.05); neither treatment nor time had an effect on LDH activity. Exposure to contaminated sediment did not have a significant effect on CS activity, however, time had a significant effect on CS activity (P<0.05). Both time and treatment had an effect on LDH activity (P<0.05). Results demonstrated that changes in gill CS and LDH are not sensitive biomarkers of petroleum hydrocarbon exposure in M. edulis planulatus.     

Comparative evaluation of biomarkers of occupational exposure to toluene

Objectives This study was initiated to make comparative evaluation of five proposed urinary markers of occupational exposure to toluene, i.e., benzyl alcohol, benzylmercapturic acid, o-cresol, hippuric acid and un-metabolized toluene. Methods In practice, six plants in Japan were surveyed, and 122 Japanese workers (mostly printers; all men) together with 12 occupationally nonexposed control subjects (to be called controls; all men) agreed to participate in the study. Surveys were conducted in the second half of working weeks. Time-weighted average exposure (about 8 h) to toluene and other solvents were monitored by diffusive sampling. End-of-shift urine samples were collected and analyzed for the five markers by the methods previously described; simultaneous determination of o-cresol was possible by the method originally developed for benzyl alcohol analysis. Results The toluene concentration in the six plants was such that the grand geometric mean (GM) for the 122 cases was 10.4 ppm with the maximum of 121 ppm. Other solvents coexposed included ethyl acetate (26 ppm as GM), methyl ethyl ketone (26 ppm), butyl acetate (1 ppm) and xylenes (1 ppm). By simple regression analysis, hippuric acid correlated most closely with toluene in air (r = 0.85 for non-corrected observed values) followed by un-metabolized toluene (r = 0.83) and o-cresol (r = 0.81). In a plant where toluene in air was low (i.e., 2 ppm as GM), however, un-metabolized toluene and benzylmercapturic acid in urine showed better correlation with air-borne toluene (r = 0.79 and 0.61, respectively) than hippuric acid (r = 0.12) or o-cresol (r = 0.17). Benzyl alcohol tended to increase only when toluene exposure was intense. Correction for creatinine concentration or specific gravity of urine did not improve the correlation in any case. Multiple regression analysis showed that solvents other than toluene did not affect the levels of o-cresol, hippuric acid or un-metabolized toluene. Levels of benzylmercapturic acid and un-metabolized toluene were below the limits of detection [limit of detections (LODs); 0.2 and 2 μg/l, respectively] in the urine from the control subjects. Conclusions In over-all evaluation, hippuric acid, followed by un-metabolized toluene and o-cresol, is the marker of choice for occupational toluene exposure. When toluene exposure level is low (e.g., 2 ppm), un-metabolized toluene and benzylmercapturic acid in urine may be better indicators. Detection of un-metabolized toluene or benzylmercapturic acid in urine at the levels in excess of the LODs may be taken as a positive evidence of toluene exposure, because their levels in urine from the controls are below the LODs. The value of benzyl alcohol as an exposure marker should be limited.     

Evaluation of urinary biomarkers of exposure to benzene: correlation with blood benzene and influence of confounding factors

Purpose   trans,trans-Muconic acid (t,t-MA) is generally considered as a useful biomarker of exposure to benzene. However, because of its lack of specificity, concerns about its value at low level of exposure have recently been raised. The aim of this study was (a) to compare t,t-MA, S-phenylmercapturic acid (SPMA) and benzene (B-U) as urinary biomarkers of exposure to low levels of benzene in petrochemical workers and, (b) to evaluate the influence of sorbic acid (SA) and genetic polymorphisms of biotransformation enzymes on the excretion of these biomarkers. Method  A total of 110 workers (including 24 smokers; 2–10 cigarettes/day) accepted to take part in the study. To assess external exposure to benzene, air samples were collected during the whole working period by a passive sampling device attached close to the breathing zone of 98 workers. Benzene was measured in blood (B-B) samples taken at the end of the shift, and was considered as the reference marker of internal dose. Urine was collected at the end of the shift for the determination of B-U, SPMA, t,t-MA, SA and creatinine (cr). B-U and B-B were determined by head-space/GC–MS, SPMA and SA by LC-MS, t,t-MA by HPLC-UV. Results  Most (89%) personal measurements of airborne benzene were below the limit of detection (0.1 ppm); B-B ranged from <0.10 to 13.58 μg/l (median 0.405 μg/l). The median (range) concentrations of the urinary biomarkers were as follows: B-U 0.27 μg/l (<0.10–5.35), t,t-MA 0.060 mg/l (<0.02–0.92), SPMA 1.40 μg/l (0.20–14.70). Urinary SA concentrations ranged between <3 and 2,211 μg/l (median 28.00). Benzene concentration in blood and in urine as well as SPMA, but not t,t-MA, were significantly higher in smokers than in non-smokers. The best correlation between B-B and urinary biomarkers of exposure were obtained with benzene in urine (μg/l r = 0.514, P < 0.001; μg/g cr r = 0.478, P < 0.001) and SPMA (μg/l r = 0.495, P < 0.001; μg/g cr r = 0.426, P < 0.001) followed by t,t-MA (mg/l r = 0.363, P < 0.001; mg/g cr r = 0.300, P = 0.002). SA and t,t-MA were highly correlated (r = 0.618, P < 0.001; corrected for cr r = 0.637). Multiple linear regression showed that the variation of t,t-MA was mostly explained by SA concentration in urine (30% of the explained variance) and by B-B (12%). Variations of SPMA and B-U were explained for 18 and 29%, respectively, by B-B. About 30% of the variance of B-U and SPMA were explained by B-B and smoking status. Genetic polymorphisms for biotransformation enzymes (CYP2E1, EPHX1, GSTM1, GSTT1, GSTP1) did not significantly influence the urinary concentration of any of the three urinary biomarkers at this low level of exposure. Conclusion  At low levels of benzene exposure (<0.1 ppm), (1) t,t-MA is definitely not a reliable biomarker of benzene exposure because of the clear influence of SA originating from food, (2) SPMA and B-U reflect the internal dose with almost similar accuracies, (3) genetically based inter-individual variability in urinary excretion of biomarkers seems negligible. It remains to assess which biomarker is the best predictor of health effects.     

Oncogene proteins as biomarkers in the molecular epidemiology of occupational carcinogenesis

Summary The use of oncogene proteins as biomarkers offers a new approach to the molecular epidemiologic evaluation of occupational carcinogenesis. The ras oncogene-encoded p21 protein represents a prototype for this type of study, since it is known to be activated by common occupational carcinogens, is frequently found in human tumors of occupational concern, and, at least in certain instances, appears to be expressed relatively early in the disease process, allowing the possibility of early detection and intervention. Herein, we review our experience with the use of immunologic detection of p21 in cohorts with cancer or at risk for the development of cancer due to their occupational exposures. The results suggest that p21 (particularly when used with other oncoproteins and biomarkers such as PAH-DNA adducts) will indeed be a useful addition to the growing armamentarium of molecular epidemiologic biomarkers in the study of occupational carcinogenic mechanisms and in the detection and prevention of occupational cancers.This paper was delivered at the presentation of the Robert R. J. Hilker Award in Occupational Medicine, Chicago, Illinois, March 30, 1990     

Laboratory investigation into the development of resistance of Daphnia magna to the herbicide molinate

Daphnia magna (F0 generation) was exposed to different sublethal molinate concentrations (0, 3.77, 4.71, 6.28, 9.42, and 18.85 mg/L) during 21 days. Chronic toxicity tests, using the same herbicide concentrations, were also carried out during 21 days using neonates of F1 first brood (F1-1st) and F1 third brood (F1-3rd) offspring generations from the parentals (F0) preexposed to the herbicide. Finally, offspring (from F1-1st and F1-3rd broods) were transferred to herbicide-free medium during a 21-day recovery period. The alga Nannochloris oculata (5 x 10(5) cells/mL) was used as food in all the experiments. The effect of molinate on survival, reproduction, and growth was monitored for the selected daphnid generations. The parameters used to evaluate herbicide effect on reproduction were mean total young per female, mean brood size, time to first reproduction, mean number broods per female, and intrinsic rate of natural increase (r). Survival and growth (body length) were also determined after 21 days of exposure to molinate. Reproduction was significantly reduced when molinate concentration was increased in the medium. This effect was higher in the parental (F0) daphnids than in the F1-1st and F1-3rd offspring. In the recovery study, reproduction was still reduced in F1 generation daphnids (1st and 3rd), but only in those animals from parentals exposed to the highest molinate concentrations. The decreased with increasing concentrations of molinate in daphnids from the parental generation (F0). Significant differences were also found in daphnids from the F1 generation (exposure). The growth of the exposed organisms (F0 and F1) decreased, although the greatest decrease was found in the parental animals (F0) (25%) exposed to 9.42 mg/L molinate. F1 daphnids (1st and 3rd broods) from the recovery period did not show any significant difference in their growth after 21 days of study. Finally, survival was not affected after exposure to the selected molinate concentrations except in those daphnids from the F0 generation in which survival decreased 51% and 78% at the highest herbicide concentrations tested (9.42 and 18.85 mg/L). Our results suggest that the offspring daphnids seem to be adaptated to the herbicide molinate, showing more longevity and reproduction than their parentals.     

Sensitivity of offspring to chronic 3,4-dichloroaniline exposure varies with maternal exposure

Many anthropogenic pollutants are present in aquatic systems for extended periods of time. Populations in the field may be exposed to toxicants for several generations, which may affect their sensitivity to toxicants. Ceriodaphnia cf. dubia mass cultures were maintained for four generations in various concentrations of 3,4-dichoroaniline (0, 2.5, 5, 10, and 15 microg/L) and were reestablished every generation using fourth-brood neonates. Each generation, chronic toxicity tests were initiated using fourth-brood neonates from each mass culture treatment. Significantly (P < 0.05) reduced sensitivity to 3,4-dichloroaniline compared to control animals was shown by F1 offspring from mothers exposed to 15 microg/L, F2 offspring from mothers exposed to 5, 10, and 15 microg/L, F3 offspring from mothers exposed to 10 and 15 microg/L, and F4 offspring from mothers exposed to all 3,4-dichloroaniline treatments (2.5, 5, 10, and 15 microg/L). Possible explanations for the development of tolerance, and the possible implications of tolerance, are discussed.     

Acute toxicity of pesticides to the tropical freshwater shrimp Caridina laevis

To determine the potential risk of pesticides frequently used in Indonesia, a new toxicity test was developed using the indigenous freshwater shrimp Caridina laevis, which is representative of tropical ecosystems. The test species could easily be maintained in the laboratory. Acute toxicity tests with different insecticides, fungicides, and herbicides assessing adult survival after 24 and 96 h of static exposure showed low control mortality (<10%) and good reproducibility for diazinon. Juvenile C. laevis appeared approximately two to three times more sensitive to diazinon than adult ones. Compared to other species of freshwater crustacean commonly used in standard aquatic toxicity tests, C. laevis showed similar sensitivity for diazinon and lambda cyhalothrin, much greater sensitivity for endosulfan and paraquat and much lower sensitivity for carbofuran. It may be concluded that the acute toxicity test using C. laevis may be a suitable alternative for determining the potential risk of chemicals under tropical conditions.     

Use of the multispecies freshwater biomonitor to assess behavioral changes of Corophium volutator (Pallas, 1766) (Crustacea, Amphipoda) in response to toxicant exposure in sediment

Automated sediment toxicity testing and biomonitoring has grown rapidly. This study tested the suitability of the marine amphipod Corophium volutator (Pallas, 1766) for sediment biomonitoring using the Multispecies Freshwater Biomonitor (MFB). Two experiments were undertaken to (1) characterize individual behaviors of C. volutator using the MFB and (2) examine behavioral changes in response to sediment spiked with the pesticide Bioban. Four behaviors were visually identified (walking, swimming, grooming and falling) and characterized in the MFB as different patterns of locomotor activity (0-2 Hz range). Ventilation was not visually observed but was detected by the MFB (2-8 Hz). No clear diel activity patterns were detected. The MFB detected an overall increase in C. volutator locomotor activity after Bioban addition to the sediments (56, 100, 121 mg kg(-1)). C. volutator was more active (both locomotion and ventilation) in the water column than the spiked sediment. C. volutator appears a sensitive and appropriate species for behavioral sediment toxicity assessment and biomonitoring.     

Toluene itself as the best urinary marker of toluene exposure

Head-space gas chromatography (GC) and high-performance liquid chromatography (HPLC) (with fluorescence detectors) methods were developed for toluene (TOL-U) and o-cresol (CR-U) in urine, respectively. In order to identify the most sensitive urinary indicator of occupational exposure to toluene vapor (TOL-A) among TOL-U, CR-U, and hippuric acid in urine (HA-U), the two methods together with an HPLC (with untraviolet detectors) method for determination of HA-U were applied in the analysis of end-of-shift urine samples from 115 solvent-exposed workers (exposed to toluene at 4 ppm as geometric mean). Regression analysis showed that TOL-U correlated with TOL-A with a significantly higher correlation coefficient than did HA-U or CR-U. With regard to the TOL-A concentrations at which the exposed subjects could be separated from the nonexposed by the analyte, TOL-U achieved separation at < 10 ppm TOL-A, whereas both HA-U and CR-U did so only when TOL-A was 30 ppm or even higher. The ratio of the analyte concentrations at 50 ppm TOL-A to those at 0 ppm TOL-A was also highest for TOL-U. Overall, the results suggest that TOL-U is a better marker of exposure to toluene vapor than HA-U or CR-U.     

Evaluation of occupational exposure to benzene by urinalysis

Urinary phenol determinations have traditionally been used to monitor high levels of occupational benzene exposure. However, urinary phenol cannot be used to monitor low-level exposures. New biological indexes for exposure to low levels of benzene are thus needed. The aim of this study was to investigate the relations between exposure to benzene (Abenzene, ppm), as measured by personal air sampling, and the excretion of benzene (U-benzene, ng/l),trans,trans-muconic acid (MA, mg/g creatinine), andS-phenylmercapturic acid (PMA, g/g creatinine) in urine. The subjects of the study were 145 workers exposed to benzene in a chemical plant. The geometric mean exposure level was 0.1 ppm (geometric standard deviation = 4.16). After logarithmic transformation of the data the following linear regressions were found: log (U-benzene, ng/l) = 0.681 log (A-benzene ppm) + 4.018; log (MA, mg/g creatinine) = 0.429 log (A-benzen ppm) – 0.304; and log (PMA, g/g creatinine) = 0.712 log (A-benzene ppm) + 1.664. The correlation coefficients were, respectively, 0.66, 0.58, and 0.74. On the basis of the equations it was possible to establish tentative biological limit values corresponding to the respective occupational exposure limit values. In conclusion, the concentrations of benzene, mercapturic acid, and muconic acid in urine proved to be good parameters for monitoring low benzene exposure at the workplace.     

Effect of the exposure to Mn-doped ZnS nanoparticles on biomarkers in the freshwater western mosquitofish Gambusia affinis

Synthesized Mn-doped ZnS nanoparticles, with 10% of Mn dopant, were used to investigate their environmental toxicity. Mn-doped ZnS quantum dots (QDs) stabilized by 3-mercaptopropionic acid (MPA) were synthesized in a basic aqueous solution using the nucleation doping. The optical properties and structure of the obtained Mn (10%):ZnS QDs have been characterized by X-ray diffraction, UV–vis, photoluminescence spectroscopies and transmission electron microscopy. The brain, gills and liver stained sections from Gambusia affinis were dissected. Antioxidant enzyme activities (acetylcholinesterase and catalase), as well as malondialdehyde and H₂O₂ levels, were determined after exposure (94 h) to 14 and 28 mg/L of nanoparticles. The obtained nearly monodisperse Mn(10%):ZnS@MPA QDs have an average diameter of ca. 2.8 nm and a zinc-blende crystal structure. Mn-doped ZnS acts differently on the activities of the biomarkers in a dose-dependent manner. The recorded alterations varied between organs. Such findings provide information on the biological target of nanoparticles and their behaviour within the environment.     

Micronucleus test in freshwater fish species: an evaluation of its sensitivity for application in field surveys

Brown trout, Salmo trutta, European eel, Anguilla anguilla, and European minnow, Phoxinus phoxinus, three fish species inhabiting European freshwater ecosystems, were evaluated for their use as in situ pollution biomarkers using the micronucleus test in renal erythrocytes. Experimental exposure (by immersion) to different concentrations of cyclophosphamide, colchicine, and cadmium showed that brown trout are more sensitive to the three compounds than minnows and eels. In situ surveys of wild freshwater ecosystems with different levels of pollution showed that minnows and eels living in polluted sites do not present higher micronuclei averages than those caught in clean rivers systems, whereas micronuclei are induced in brown trout inhabiting polluted sites. Our results demonstrated the suitability of brown trout for in situ biomonitoring of freshwater ecosystems as well as for laboratory tests using the micronucleus test.     

The method of choice for the determination of 2,5-hexanedione as an indicator of occupational exposure to n-hexane

Summary To identify the method of choice for analysis of urine for 2,5-hexanedione (2,5-HD) as an indicator of occupational exposure to n-hexane, the end-of-shift urine samples of 36 n-hexane exposed male workers and 30 non-exposed male workers were analyzed for 2,5-HD under three conditions of hydrolysis, i.e. enzymic hydrolysis at pH 4.8, acid hydrolysis at pH 0.5, and without hydrolysis. The 2,5-HD concentrations thus determined were examined for correlation with 8-h, time-weighted average exposure concentrations of n-hexane measured by diffusive sampling. The regression analysis showed that the 2,5-HD concentrations without any hydrolysis correlated best with the intensity of exposure to n-hexane. No 2,5-HD was detected in the urine of the non-exposed subjects under the analytical conditions with no hydrolysis. Thus, the analysis without hydrolysis was considered to be the method of choice from the viewpoint of simplicity in analytical procedures, sensitive separation of the exposed from the non-exposed, and quantitative increase in the amount of 2,5-HD after n-hexane exposure.A part of this work was presented at the 63rd Annual Meeting of Japan Association of Industrial Health, held in Kumamoto, Japan, on 3rd–6th April, 1990     

Effects of five rice herbicides on the growth of two threatened aquatic ferns

The effects of five rice herbicides bensulfuron methyl, mefenacet, quinoclamine, simetryn, and thiobencarb on the growth of two threatened aquatic ferns Azolla japonica and Salvinia natans were tested using 12-day exposure experiments at 0.1-100 nM which are expected to be present in drainages and rivers in Japan. As a reference species, Lenma minor was also used to examine the toxicity of bensulfuron methyl. Bensulfuron methyl had the most pronounced effect on the relative growth rate (RGR) of A. japonica, S. natans, and L. minor with an EC50 of 5.0, 0.54, and 10 nM, respectively. The other herbicides reduced the RGR of the aquatic ferns only at the highest concentration (100 nM) or not at all. S. natans showed the highest susceptibility to bensulfuron methyl among the three species, and the EC50 for this species was comparable to or below the maximum concentration (0.49-5.6 nM) that had been previously detected in 7 of 11 rivers in Japan. These results suggest that bensulfuron methyl runoff in drainages and rivers in Japan is expected to have adverse effects on the growth of threatened aquatic ferns in some cases, and that no or small effects occur for the other four herbicides tested.     

Using native riparian barriers to reduce Giardia in agricultural runoff to freshwater ecosystems

Waterway degradation in agricultural settings is caused by direct and diffuse sources of pollution. Waterway fencing focuses on reducing direct faecal contamination, but the extent to which it reduces overland surface runoff of pathogens is unknown. This study evaluated the potential of four riparian treatments to reduce Giardia in saturation excess surface runoff entering the waterway. Treatment 1 comprised exotic pasture grass and weeds that regenerated from bare soil between the fence and the waterway in the absence of cattle grazing and was compared with three others comprising monocultural plantings of New Zealand native grassland plants. Runoff experiments involving Giardia were performed after planting, both prior to and following the summer growing season. Giardia was not detected from any plot prior to cyst addition. In spring the native ‘C. secta’, ‘A. lessoniana’ and ‘C. richardii’ treatments showed significantly greater reductions in Giardia in runoff than the ‘exotic grasses’ treatment, while in autumn the ‘C. richardii’ treatment reduced Giardia more than the ‘exotic grasses/weeds’. A reduction in public health risk should follow from riparian vegetation, whether exotic or native, but with an added benefit in the case of the native tussock grass C. richardii, due to the associated lower runoff rate.     

Comparative evaluation of urinalysis and blood analysis as means of detecting exposure to organic solvents at low concentrations

Summary One hundred and forty-three workers exposed to one or more of toluene, xylene, ethylbenzene, styrene, n-hexane, and methanol at sub-occupational exposure limits were examined for the time-weighted average intensity of exposure by diffusive sampling, and for biological exposure indicators by means of analysis of shift-end blood for the solvent and analysis of shift-end urine for the corresponding metabolite(s). Urinalysis was also performed in 20 nonexposed control men to establish the background level. Both solvent concentrations in blood and metabolite concentrations in urine correlated significantly with solvent concentrations in air. Comparison of blood analysis and urinalysis as regards sensitivity in identifying low solvent exposure showed that blood analysis is generally superior to urinalysis. It was also noted that estimation of exposure intensity on an individual basis is scarcely possible even with blood analysis. Solvent concentration in whole blood was the same as that in serum in the case of the aromatics, except for styrene. It was higher in blood than in serum in the case of n-hexane, and lower in the cases of styrene and methanol.     

Low level occupational exposure to styrene: its effects on DNA damage and DNA repair

The present study aimed to evaluate the effects of styrene exposure at levels below the recommended standards of the Threshold Limit Value (TLV-TWA(8)) of 20 ppm (ACGIH, 2004) in reinforced-fiberglass plastics workers. Study subjects comprised 50 exposed workers and 40 control subjects. The exposed workers were stratified by styrene exposure levels, i.e. group I (<10 ppm, <42.20 mg/m(3)), group II (10-20 ppm, 42.20-84.40 mg/m(3)), and group III (>20 ppm, >84.40 mg/m(3)). The mean styrene exposure levels of exposed workers were significantly higher than those of the control workers. Biomarkers of exposure to styrene, including blood styrene and the urinary metabolites, mandelic acid (MA) and phenylglyoxylic acid (PGA), were significantly increased with increasing levels of styrene exposure, but were not detected in the control group. DNA damage, such as DNA strand breaks, 8-hydroxydeoxyguanosine (8-OHdG), and DNA repair capacity, were used as biomarkers of early biological effects. DNA strand breaks and 8-OHdG/10(5)dG levels in peripheral leukocytes of exposed groups were significantly higher compared to the control group (P<0.05). In addition, DNA repair capacity, determined by the cytogenetic challenge assay, was lower in all exposed groups when compared to the control group (P<0.05). The expression of CYP2E1, which is involved in styrene metabolism, in all styrene exposed groups, was higher than that of the control group at a statistically significant level (P<0.05). Levels of expression of the DNA repair genes hOGG1 and XRCC1 were significantly higher in all exposed groups than in the control group (P<0.05). In addition to styrene contamination in ambient air, a trace amount of benzene was also found but, the correlation between benzene exposure and DNA damage or DNA repair capacity was not statistically significant. The results obtained from this study indicate an increase in genotoxic effects and thus health risk from occupational styrene exposure, even at levels below the recommended TLV-TWA(8) of 20 ppm.     

Biological monitoring of exposure to solvents using the chemical itself in urine: application to toluene

Objective Biomonitoring of solvents using the unchanged substance in urine as exposure indicator is still relatively scarce due to some discrepancies between the results reported in the literature. Based on the assessment of toluene exposure, the aim of this work was to evaluate the effects of some steps likely to bias the results and to measure urinary toluene both in volunteers experimentally exposed and in workers of rotogravure factories. Methods Static headspace was used for toluene analysis. o-Cresol was also measured for comparison. Urine collection, storage and conservation conditions were studied to evaluate possible loss or contamination of toluene in controlled situations applied to six volunteers in an exposure chamber according to four scenarios with exposure at stable levels from 10 to 50 ppm. Kinetics of elimination of toluene were determined over 24 h. A field study was then carried out in a total of 29 workers from two rotogravure printing facilities. Results Potential contamination during urine collection in the field is confirmed to be a real problem but technical precautions for sampling, storage and analysis can be easily followed to control the situation. In the volunteers at rest, urinary toluene showed a rapid increase after 2 h with a steady level after about 3 h. At 47.1 ppm the mean cumulated excretion was about 0.005% of the amount of the toluene ventilated. Correlation between the toluene levels in air and in end of exposure urinary sample was excellent (r = 0.965). In the field study, the median personal exposure to toluene was 32 ppm (range 3.6–148). According to the correlations between environmental and biological monitoring data, the post-shift urinary toluene (r = 0.921) and o-cresol (r = 0.873) concentrations were, respectively, 75.6 μg/l and 0.76 mg/g creatinine for 50 ppm toluene personal exposure. The corresponding urinary toluene concentration before the next shift was 11 μg/l (r = 0.883). Conclusion Urinary toluene was shown once more time a very interesting surrogate to o-cresol and could be recommended as a biomarker of choice for solvent exposure.     

Tolerance of the lichen Xanthoria parietina (L.) Th. Fr. to metal stress

Comparisons were made between the content of Al, Cd, Cu, Fe, Mn, Pb, Sb and Zn in thalli of the lichen Xanthoria parietina and selected physiological processes. Assimilation pigments, activity of photosystem II, parietin, reactive substances produced using thiobarbituric acid (TBA) and CO₂ gas exchange were measured in order to assess tolerance to atmospheric sources of pollution. As expected, lichen thalli accumulated high amounts of the measured elements in relation to distance from pollution sources in Košice, Slovak Republic (US Steel factory and vehicular traffic in the city center). However, except for TBA reactive substances production and to some extent CO₂ gas exchange at the most polluted station, none of the tested physiological parameters showed a clear correlation between accumulation of elements and physiological damage. This reflected a high degree of pollution tolerance in this lichen and corresponded with its high abundance in Košice.