Genotoxic damage in free-living Algerian mouse (Mus spretus) after the Coto Doñana ecological disaster

The Do?ana National Park (Spain), one of the most important wildlife sites in the West of Europe, was affected (25 April 1998) by the spill of acidic waste rich in toxic metals (mainly zinc, lead, copper, etc.), arsenic and aromatic amines from the Aznalcollar mine accident. Micronuclei test with May Grunwald-Giemsa and with CREST-antikinetocore staining using DAPI as counter-staining were performed on peripheral blood erythrocytes from Algerian mice to evaluate genotoxic damage. Animals were collected in four locations each differently affected by the disaster. Higher frequencies of micronuclei and CREST-positive micronuclei were observed in the sites, which were reached by toxic sludge and contaminated water in comparison with those located within the park. The results obtained applying the two methods indicate that DAPI staining is more sensitive in detecting micronuclei. Genotoxic biomonitoring should be further carried out in the area to control the mutagenetic level in natural populations.     

Micronucleus assay and lymphocyte mitotic activity in risk assessment of occupational exposure to microwave radiation

The effects of radiofrequency electromagnetic radiation (RFR) on the cell kinetics and genome damages in peripheral blood lymphocytes were determined in lymphocytes of 12 subjects occupationally exposed to microwave radiation. Results showed an increase in frequency of micronuclei (MN) as well as disturbances in the distribution of cells over the first, second and third mitotic division in exposed subjects compared to controls. According to previous reports micronucleus assay can serve as a suitable indicator for the assessment of exposure to genotoxic agents (such as RFR) and the analysis of mitotic activity as an additional parameter for the efficient biomonitoring.     

Genotoxicity evaluation of the insecticide endosulfan in the wetland macrophyte Bidens laevis L

The frequency of micronuclei (MN) and chromosome aberrations in anaphase-telophase (CAAT) was determined in root tips of the wetland macrophyte Bidens laevis exposed to environmentally relevant concentrations of endosulfan (0.01, 0.02, 0.5 and 5microg/L) for 48h. MN frequency varied from 0 in negative controls and plants exposed to 0.01microg/L endosulfan to 0-3 in plants exposed to 5microg/L. Moreover, a significant concentration-dependent increase of CAAT was observed. The higher proportion of laggards and vagrand chromosomes observed at 5microg/L would indicate that endosulfan interacts with the spindle interrupting normal chromosome migration. Endosulfan resulted genotoxic to B. laevis, a species of potential value for bioassays and in situ monitoring of environmental contamination by pesticides.     

Genotoxicity of two novel pesticides for the earthworm, Eisenia fetida

In this paper, several studies were conducted to evaluate the genotoxicity of two pesticides, Imidacloprid and RH-5849, for earthworm (Eisenia fetida). Earthworms were exposed in different exposure systems to evaluate their acute toxicity and the genotoxicity of the two pesticides was evaluated by using the method of sperm deformity assessment, micronucleus test of root tip cells in Vicia faba, a mouse bone-marrow micronucleus test, and comet assay. LC(50) (interpolated concentration at which 50% mortality of test population occurs) for earthworms varied in different exposure systems. The results indicated that Imidacloprid was consistently more toxic than RH-5849 in all exposure systems. In this study, sperm deformity test was used to detect the potential adverse influences of pesticides on the reproduction of earthworms. The results demonstrated that significant induction of sperm deformity (p<0.01) and a dose-effect relationship displayed at Imidacloprid concentrations higher than 0.5 mg/kg dry soil. However, the sperm deformity frequency of groups exposed to RH-5849 did not show significant difference (p>0.05) from the control until the dose reached 100 mg/kg dry soil. The results of the V. faba micronucleus tests showed that micronuclei frequency of the exposed group did not show significant difference (p>0.05) from the control until the concentration of Imidacloprid and RH-5849 reached 100 mg/ml. The results of the mouse bone-marrow micronuclei test also indicate that two pesticides did not show significant effects (p>0.05) on the micronuclei frequency in mice bone-marrow cells until the dose reached 100 mg/kg for Imidacloprid and 300 mg/kg for RH-5849 (2/3 LD(50)). Although no genotoxicity was detected by using the micronucleus tests, the results of the comet assay showed that the two pesticides induce significant DNA damage (p<0.01) in earthworms and dose-effect relationships were displayed. The 'earthworm comet assay' is a rapid and sensitive way to screen chemicals or terrestrial environments for their DNA-damaging properties.     

Effects of triclosan on zebrafish early-life stages and adults

Background, aim and scope  

The biocide triclosan (TCS) is commonly used in personal care, acrylic, plastic, and textiles products. TCS has been detected in surface water in several countries, and its ecological impact is largely unknown. In this work, the toxicity of TCS in zebrafish (Danio rerio), embryos and adults was studied. Several lethal and sub-lethal endpoints were analysed in organisms exposed to TCS such as mortality, embryo development and behaviour, hatching, micronuclei and biochemical markers (cholinesterase (ChE), glutathione S-transferase (GST) and lactate dehydrogenase (LDH)).     

Cytogenetic investigation of women exposed to different levels of dioxins in Chapaevsk town

Three groups of women (aged 20-40 years) exposed to different levels of dioxins were studied in Chapaevsk town: 15 women working at the chemical fertilizer plant where occupational exposure to dioxins is possible; 16 women without dioxins occupational exposure, but living as far as 1-3 km from the plant; 14 women without dioxins occupational exposure and living as far as 5-8 km from the plant. No personal correlation related to dioxins exposure was found by chromosome aberrations (CA) in peripheral blood lymphocytes, micronuclei (MN) and nuclear anomalies in buccal mucosa cells. There were no significant differences between the groups in CA and MN. Karyopyknosis and karyorrhexis were significantly increased in the highest exposed group.     

Cell cycle stage specific application of municipal landfill leachates to assess the genotoxicity in root meristem cells of barley (Hordeum vulgare)

Municipal solid wastes (MSW) are unavoidable sources of environmental pollution. Improper disposal of municipal waste results in the leaching of toxic metals and organic chemicals, which can contaminate the surface and ground water leading to serious health hazard. In this study, the toxic effects of the leachate prepared from municipal solid waste samples were examined in root meristem cells of barley (Hordeum vulgare L.) at various stages of cell cycle, i.e., G₁, S, and G₂. Seeds of barley were exposed to 2.5, 5, and 10 % of leachates in soil and aqueous media in 48 h at different cell cycle stages. The physicochemical data of the present study revealed that municipal solid waste leachate contains high amount of heavy metals, which significantly affected growth and physiological activities of barley. Significant inhibition in hypocotyl length, germination, and mitotic index were observed at all concentration of leachate treatment. Induction of chromosomal aberrations (CA’s) and micronuclei (MN) formation were also observed with different concentrations of leachate treatment at 7, 17, and 27 h of presoaking durations, which falls in G₁, S, and G₂ phase of the cell cycle, respectively. Also, exposure of leachate at S phase of the cell cycle had significant effects in barley through chromosomal aberration and micronuclei formation.     

Aluminum binding to ion exchange sites in acid-sensitive versus acid-tolerant cladocerans

Two acid-sensitive cladocerans, Daphnia galeata mendotae and D. retrocurva, and one acid-tolerant one, Bosmina longirostris, were exposed for 24 h to pH 5.0 and 200 microg liter(-1) total Al. The entire procedure was replicated on three dates in summer 1989. Mortality rates were determined, and the extent of Al binding to ion exchange sites determined using hematoxylin staining. Both daphnids consistently experienced near 100% mortalities, while mortalities for B. longirostris were always near zero. The daphnids showed marked Al binding at the maxillary glands, the site of ion exchange, while B. longirostris showed no noticeable Al binding.     

Studies on the ability of water hyacinth (Eichhornia crassipes) to bioconcentrate and biomonitor aquatic mercury

Water hyacinth (Eichhornia crassipes, Mart solms) plants were employed to assess bioconcentration and genotoxicity of aquatic mercury. Plants were exposed to water contaminated with mercuric chloride (MC) or phenyl mercuric acetate (PMA) at 0.001 to 1.0 mg litre(-1), or mercury contaminated effluent from a chloralkali plant for various periods of 4 t0 96 h. Root samples taken after 4, 8, 12, 24, 48, 72 and 96 h of exposure were analysed for bioconcentration of mercury spectrophotometrically, and the root meristems were fixed in aceto-ethanol for cytological analysis to determine the frequencies of cells with micronuclei (MNC). Ethyl methane sulfonate and tap water served as positive and negative controls, respectively. The results indicated that bioconcentration of mercury in root tissue was both time- and concentration-dependent, providing evidence that water hyacinth is a good absorbant of aquatic mercury. The frequency of root meristematic cells with MNC followed a concentration-response. The findings indicate the potential of water hyacinth plants for in situ monitoring and for mitigation of aquatic mercury pollution.     

Assessment of the relationship between total suspended particles and the response of two biological indicators transplanted to an urban area in central Argentina

Samples of the vascular plant Tradescantia pallida and the lichen Usnea amblyoclada were exposed from October 2004 to April 2005 in three sites with different local sources of air pollution in Córdoba city, Argentina. Simultaneous determinations of the ambient levels of total suspended particles were made for each site. Young inflorescenses of T. pallida were collected in November, February and April and the frequency of micronuclei was determined on early tetrads of pollen mother cells. Physiological parameters and the elemental composition of lichen thalli were measured from samples exposed and replaced every month. Significant differences among sampling sites were observed in the frequency of micronuclei measured in T. pallida as well as in many physiological parameters and elements accumulated in lichen thalli. The mass of particulate material as well as the concentration of Ca, Mn, Cu, Zn and Sr was significantly different in different sampling sites, too. These results suggest that in situ biomonitoring using both higher plants and lichens may be of use to characterize air pollution in areas devoid of instrumental monitoring techniques or where it is necessary to explore the distribution of air contaminants at a microscale.     

In situ monitoring of urban air in Southern Italy with the Tradescantia micronucleus bioassay and semipermeable membrane devices (SPMDs)

This study was designed to assess toxic and genotoxic compounds in the urban air of Caserta, South Italy using cuttings from the plant Tradescantia #4430. In situ monitoring of gaseous pollutans was made at 17 sampling points in two seasons of the year. Genotoxicity was evaluated by recording the micronuclei in meiotic pollen mother cells (Trad-MCN assay). In addition, the passive sampler semipermeable membrane devices (SPMDs) were deployed at the sampling points with a significant increase in micronuclei frequency. SPMDs concentrated priority organic pollutants were identified by high performance liquid chromatography and gas chromatography, while toxicity and mutagenesis were assessed on the bacterium Vibrio fischeri using the Microtox and Mutatox systems respectively. Significant toxic and mutagenic effects were observed at different points on the town grid and SPMDs effectively concentrated trace contaminants. The relationship between what was present in the air sampled by SPMDs and the micronuclei frequency was also explored.     

Association between micronuclei frequency in pollen mother cells of Tradescantia and mortality due to cancer and cardiovascular diseases: A preliminary study in Sao José dos Campos, Brazil

The present study was designed to explore the correlation between the frequency of micronuclei in Trad-MN, measured across 28 biomonitoring stations during the period comprised between 11 of May and 2 of October, 2006, and adjusted mortality rates due to cardiovascular, respiratory diseases and cancer in Sao José dos Campos, Brazil, an area with different sources of air pollution. For controlling purposes, mortality rate due to gastrointestinal diseases (an event less prone to be affected by air pollution) was also considered in the analysis. Spatial distribution of micronuclei frequency was determined using average interpolation. The association between health estimators and micronuclei frequency was determined by measures of Pearson's correlation. Higher frequencies of micronuclei were detected in areas with high traffic and close to a petrochemical pole. Significant associations were detected between micronuclei frequency and adjusted mortality rate due to cardiovascular diseases (r = 0.841, p = 0.036) and cancer (r = 0.890, p = 0.018). The association between mortality due to chronic obstructive pulmonary diseases was positive but did not reach statistical significance (r = 0.640, p = 0.172), probably because of the small number of events. Gastrointestinal mortality did not exhibit significant association with micronuclei frequency. Because the small number of observations and the nature of an ecological study, the present findings must be considered with caution and considered as preliminary. Further studies, performed in different conditions of contamination and climate should be done before considering Trad-MN in the evaluation of human health risk imposed by air pollutants.     

Increased urinary 8-hydroxy-2��-deoxyguanosine levels in workers exposed to di-(2-ethylhexyl) phthalate in a waste plastic recycling site in China

Background

Di-(2-ethylhexyl) phthalate (DEHP) is a common plasticizer used in industrial and diverse consumer products. Animal studies indicate DEHP caused developmental, reproductive, and hepatic toxicities. However, human studies of the potential effects of DEHP are limited.

Methods

The exposed site with a history of over 20 years of waste plastic recycling was located in Hunan Province, China. The reference site without known DEHP pollution source was about 50 km far away from the exposed site. In this study, 181 workers working in plastic waste recycling and 160 gender?Cage matched farmers were recruited. DEHP concentrations in water and cultivated soil samples, serum thyroid-stimulating hormone, malondialdehyde (MDA), superoxide dismutase (SOD), urinary 8-hydroxy-2??-deoxyguanosine (8-OHdG), and micronuclei frequency in human capillary blood lymphocytes were analyzed.

Results

Mean levels of DEHP were greater in environment at the recycling site than at reference site (industry wastewater for the exposed: 42.43 ??g/l; well water: 14.20 vs. 0.79 ??g/l, pond water: 135.68 vs. 0.37 ??g/l, cultivated soil: 13.07 vs. 0.81 mg/kg, p?p?p?Conclusions The occupational DEHP exposure might contribute to oxidative deoxyribonucleic acid damage in the male workers.     

Genotoxicity and histological alterations in grey mullet Mugil liza exposed to petroleum water-soluble fraction (PWSF)

Petroleum hydrocarbons are considered one of the main organic chemicals found in water bodies. In the present study, the median lethal concentration (LC₅₀) was estimated for mullet Mugil liza after acute exposure to petroleum water-soluble fraction (PWSF). Furthermore, histopathological studies and micronuclei frequency were also performed in order to observe deleterious effects of medium-term exposition to PWSF. Mullets (25?±?2.3 g) were exposed to chronic concentrations (1.7, 3.5 and 7 % of PWSF), plus the control group, for 14 and 7 days of clearance time. Throughout the experimental period (1, 4, 14 and 21 days), blood samples were collected for analysis of micronucleus (MN) and liver and gills for histopathological study. For these procedures, seven fish were sampled per concentration tested. The LC₅₀-96 h was estimated at 37.5 % of the PWSF. The time required for MN induction was 96 h of exposure. The time of clearance was sufficient to achieve a MN frequency similar to that of the control group. Histopathological studies showed severe changes in the gill and liver tissues. The most relevant histopathology in the gills was telangiectasia. Hepatic histopathology such as cholestasis, dilated sinusoids and inflammatory infiltrates were commonly described. The MN test and histological study effectively detected damages caused by medium-term exposition to the PWSF, and despite the toxicity, a few days without exposure can minimize PWSF genotoxicity in juveniles of M. liza.     

Influence of GSTM1 and GSTT1 genotypes and confounding factors on the frequency of sister chromatid exchange and micronucleus among road construction workers

In the present study, we have investigated the influence of polymorphism of GSTM1 and GSTT1 genes and confounding factors such as age, sex, exposure duration and consumption habits on cytogenetic biomarkers. Frequency of sister chromatid exchanges (SCEs), high frequency cell (HFC) and cytokinesis blocked micronuclei (CBMN) were evaluated in peripheral blood lymphocytes of 115 occupationally exposed road construction workers and 105 unexposed individuals. The distribution of null and positive genotypes of glutathione-S transferase gene was evaluated by multiplex PCR among control and exposed subjects. An increased frequency of CBMN (7.03 ± 2.08); SCE (6.95 ± 1.76) and HFC (6.28 ± 1.69) were found in exposed subjects when compared to referent (CBMN - 3.35 ± 1.10; SCE - 4.13 ± 1.30 and HFC - 3.98 ± 1.56). These results were found statistically significant at p < 0.05. When the effect of confounding factors on the frequency of studied biomarkers was evaluated, a strong positive interaction was found. The individuals having GSTM1 and GSTT1 null genotypes had higher frequency of CBMN, SCE and HFC. The association between GSTM1 and GSTT1 genotypes and studied biomarkers was found statistically significant at p < 0.05. Our findings suggest that individuals having null type of GST are more susceptible to cytogenetic damage by occupational exposure regardless of confounding factors. There is a significant effect of polymorphism of these genes on cytogenetic biomarkers which are considered as early effects of genotoxic carcinogens.     

Volatile organohalogen compounds in human urine: the effect of environmental exposure

The paper presents the results of determination of volatile organohalogen compounds (VOX) in urine samples from subjects exposed to these compounds in their workplaces and through consumption of chlorinated tap water. The analytes were isolated and preconcentrated from the complex urine samples using the thin layer headspace (TLHS) technique with autogenous generation of the liquid sorbent. Final gas chromatographic determination was carried out by direct aqueous injection with electron capture detection (DAI-ECD). The results indicate that only a small fraction (<4%) of the VOX input is excreted with urine in the non-metabolized form. A positive correlation was found between the occupational levels of VOX in the workplace and their levels in urine. VOX levels in the urine of subjects not exposed to them in the workplace were significantly lower. Their presence in the organisms was most probably related to consumption of tap water produced by chlorination of surface waters.     

An attempt to assess the relevance of flood events—biomarker response of rainbow trout exposed to resuspended natural sediments in an annular flume

There is a consensus within the scientific community that sediments act as a long-term sink for a variety of organic and inorganic pollutants, which, however, can re-enter the water column upon resuspension of deposited material under certain hydraulic conditions such as flood events. Within the implementation of the European Water Framework Directive, it is important to understand the potential short- and long-term impact of suspended particulate matter (SPM)-associated contaminants on aquatic organisms as well as the related uptake mechanisms for a sound risk assessment. To elucidate the effects of sediment-bound organic pollutants, such as polycyclic aromatic hydrocarbons (PAHs), rainbow trout (Oncorhynchus mykiss) were exposed to three resuspended natural sediments with different contamination levels. Physicochemical parameters including dissolved oxygen concentration, pH and temperature, total PAH concentration in sediments and SPM as well as different biomarkers of exposure in fish (7-ethoxyresorufin O-deethylase activity, biliary PAH metabolites, micronuclei, and lipid peroxidation) were measured following seven days of exposure within an annular flume, a device to assess erosion and deposition processes of cohesive sediment. Concentrations of PAHs in SPM remained constant and represented the different contamination levels in the un-suspended sediments. Significant differences in bile metabolite concentrations as well as in 7-ethoxyresorufin O-deethylase induction compared to control experiments (untreated animals and animals that were exposed in the annular flume without sediment) were observed for all exposure scenarios. The ratio between 1-hydroxypyrene in bile from fish exposed to the three different contamination levels was 1.0:3.6:10.7 and correlated well with (1) the ratio of pyrene concentrations in corresponding sediments which was 1.0:3.1:12.7 and (2) with the ratio of particle-bound pyrene in SPM which was 1.0:2.7:11.7. In contrast, hepatic lipid peroxidation and micronuclei formation represented the different contamination levels less conclusive. The results of this study clearly demonstrate that firmly bound PAH from aged sediments can become bioaccessible upon resuspension under flood-like conditions and are readily absorbed by aquatic organisms such as rainbow trout. Associated short-term effects were clearly documented and possible adverse long-term impacts due to genotoxicity are likely to follow.     

Sister chromatid exchange and proliferative rate index in the longitudinal risk assessment of occupational exposure to pesticides

At present, there are more than 1,000 chemicals classified as pesticides and many reports have shown that some of them have genotoxic properties. In the present longitudinal study, possible genetic damage on a population of workers occupationally exposed to a mixture of pesticides by using sister chromatid exchange (SCE) analysis has been evaluated. As an additional cytogenetic parameter, the proportion of lymphocytes that undergo one, two or three cell divisions as well as proliferative rate index have been determined. This study was performed on the exposed group of workers employed in pesticide production, simultaneously exposed to a complex mixture of pesticides (atrazine, alachlor, cyanazine, 2,4-dichlorophenoxyacetic acid, and malathion). The blood samples of the exposed subjects were collected in three different periods: before the beginning of the new pesticide production period, after 8 months of everyday work in the pesticide production, and 8 months after the removal of subjects out of the production. In all three samplings, the mean value of SCE and number of cells with high sister chromatid exchange frequency (HFC) in the exposed group was significantly higher in the comparison with the control group. There were no differences in the proliferative rate index (PRI) between the control and exposed group, regardless of the sampling period. In both groups examined, the majority of lymphocytes were found in the second cell division, following cultivation. These results suggest that the increase in the number of SCE found in the exposed subjects is not the result of either cytotoxic or epigenetic action of pesticide mixture, but chronic occupational exposure to mixture of pesticides.     

Assessment of environmental tobacco smoke and respirable suspended particle exposures for nonsmokers in Basel by personal monitoring

One hundred and ninety-six randomly selected nonsmoking subjects collected air samples close to their breathing zone by wearing personal monitors for 24 h. The study was centred in Basel, Switzerland, and comprised housewives in one group, primarily for assessing exposures in the home, and office workers in a second group to assess the contribution of the workplace to overall exposure. Samples collected were analysed for respirable suspended particles (RSP), nicotine, 3-ethenylpyridine and environmental tobacco smoke (ETS) particles by using ultraviolet absorbance, fluorescence and solanesol measurements. Saliva cotinine analyses were also undertaken to confirm the nonsmoking status of the subjects. Based upon median 24 h time weighted average concentrations, office workers who live and work with smokers were exposed to 39 μg m^-3 RSP, 6.6 μg m^-3 ETS particles and 0.90 μg m^-3 nicotine. Housewives living with smokers were exposed to median concentrations of 34 μg m^-3 RSP, 1.4 μg m^-3 ETS particles and 0.60 μg m^-3 nicotine. Workplaces where smoking occurred were estimated, on average, to contribute between 34 and 46% to annual exposure of ETS particles and nicotine. Based upon 90th percentile values the most highly exposed housewives, those living with smokers, would potentially inhale 18 cigarette equivalents per year whilst the most highly exposed office workers, both living and working with smokers, might inhale 61 cigarette equivalents. The rate at which subjects misreported their nonsmoking status varied between 9.7 and 12.2%.     

Assessing the genotoxicity of urban air pollutants using two in situ plant bioassays

Genotoxicity of urban air has been analysed almost exclusively in airborne particulates. We monitored the genotoxic effects of airborne pollutants in the urban air of Perugia (Central Italy). Two plant bioindicators with different genetic endpoints were used: micronuclei in meiotic pollen mother cells using Tradescantia-micronucleus bioassay (Trad-MCN) and DNA damage in nuclei of Nicotiana tabacum leaves using comet assay (Nicotiana-comet). Buds of Tradescantia clone # 4430 and young N. tabacum cv. Xanthi plants were exposed for 24 h at three sites with different pollution levels. One control site (indoor control) was also used. The two bioassays showed different sensitivities toward urban pollutants: Trad-MCN assay was the most sensitive, but DNA damage in N. tabacum showed a better correlation with the pollutant concentrations. In situ biomonitoring of airborne genotoxins using higher plants combined with chemical analysis is thus recommended for characterizing genotoxicity of urban air.