Genotoxic potential of selected cytostatic drugs in human and zebrafish cells

Due to their increasing use, the residues of anti-neoplastic drugs have become emerging pollutants in aquatic environments. Most of them directly or indirectly interfere with the cell’s genome, which classifies them into a group of particularly dangerous compounds. The aim of the present study was to conduct a comparative in vitro toxicological characterisation of three commonly used cytostatics with different mechanisms of action (5-fluorouracil [5-FU], cisplatin [CDDP] and etoposide [ET]) towards zebrafish liver (ZFL) cell line, human hepatoma (HepG2) cells and human peripheral blood lymphocytes (HPBLs). Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and acridine orange/ethidium bromide staining. All three drugs induced time- and dose-dependent decreases in cell viability. The sensitivity of ZFL and HepG2 cells towards the cytotoxicity of 5-FU was comparable (half maximal inhibitory concentration (IC₅₀) 5.3 to 10.4 μg/mL). ZFL cells were more sensitive towards ET- (IC₅₀ 0.4 μg/mL) and HepG2 towards CDDP- (IC₅₀ 1.4 μg/mL) induced cytotoxicity. Genotoxicity was determined by comet assay and cytokinesis block micronucleus (CBMN) assay. ZFL cells were the most sensitive, and HPBLs were the least sensitive. In ZFL cells, induction of DNA strand breaks was a more sensitive genotoxicity endpoint than micronuclei (MNi) induction; the lowest effective concentration (LOEC) for DNA strand break induction was 0.001 μg/mL for ET, 0.01 μg/mL for 5-FU and 0.1 μg/mL for CDDP. In HepG2 cells, MNi induction was a more sensitive genotoxicity endpoint. The LOEC values were 0.01 μg/mL for ET, 0.1 μg/mL for 5-FU and 1 μg/mL for CDDP. The higher sensitivity of ZFL cells to cytostatic drugs raises the question of the impact of such compounds in aquatic ecosystem. Since little is known on the effect of such drugs on aquatic organisms, our results demonstrate that ZFL cells provide a relevant and sensitive tool to screen genotoxic potential of environmental pollutant in the frame of hazard assessment.     

Effect-based assessment of passive air samples from four countries in Eastern Europe

Although passive sampling has been previously used for the monitoring of volatile and semi-volatile contaminants in air, there are limited data on the use of this technique coupled with bioassays based on specific biological responses. Biological responses including those mediated by the aryl hydrocarbon (AhR) receptor as well as (anti-)estrogenicity and (anti-)androgenicity of samples from four Eastern European countries (Lithuania, Slovakia, Romania, and Serbia) were determined. To address the potential differences of specific toxic potencies of pollutant mixtures in ambient air in Eastern Europe, each country was characterized by a single more remote location that served to determine regional background conditions and one location in more urbanized and industrialized locations, which were defined as “impacted” areas. Besides samples from Lithuania, a significant gradient in concentrations of AhR-mediated potency from background and impacted localities was observed. Greatest potencies were measured in samples from impacted locations in Romania and Slovakia. Concentrations of polycyclic aromatic hydrocarbons (PAHs) that were quantified accounted for 3–33 % of the 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents determined by use of the bioassay. No significant estrogenic potency was detected but anti-estrogenic effects were produced by air from two background locations (Lithuania, Slovakia) and three impacted locations (Lithuania, Romania, and Serbia). Anti-androgenic potency was observed in all samples. The greatest anti-estrogenic potency was observed at the background location in Slovakia. Anti-estrogenic and anti-androgenic potencies of studied air samples were probably associated with compounds that are not routinely monitored. The study documents suitability of passive air sampling for the assessment of specific toxic potencies of ambient air pollutants.     

Impact and ecosystem service of forest and sacred grove as saviour of water quantity and quality in Garhwal Himalaya,India

The present study was conducted in environs of the sacred grove of Garhwal Himalaya, India, with a view to assess the impacts of sacred groves and forests on the quality and quantity of water and also to assess the effect of seasonality on perennial stream quality. Water samples were collected from three randomly selected stream spots of both the sacred grove dominated by deodar (Cedrus deodara) and the non-sacred patch dominated by oak (Quercus leucotrichophora). Water samples from both patches were within the World Health Organization (WHO) standard limits. Based on an already established water quality index, water quality of both patches was safe for domestic and irrigation purposes but needs treatment for drinking purposes. Results of the present study also showed a very prominent impact of forest type as well as management condition on water quality and quantity. The water discharge from an oak forest shows more consistency than the discharge from a deodar forest. Due to the presence of the sacred grove, the area has become the source of good quality water supply during lean season for the surrounding villages. Water quality and quantity differed along with the change in season. The sacred grove and the existing forest leave a great impression on local dwellers, as due to its presence, local dwellers never run out of water supply during the dry season. As a result, the villagers sincerely want to protect the area for the sake of their own well-being.     

Using historical ecology to reassess the conservation status of coniferous forests in Central Europe

Forests cover approximately one‐third of Central Europe. Oak (Quercus) and European beech (Fagus sylvatica) are considered the natural dominants at low and middle elevations, respectively. Many coniferous forests (especially of Picea abies) occur primarily at midelevations, but these are thought to have resulted from forestry plantations planted over the past 200 years. Nature conservation and forestry policy seek to promote broadleaved trees over conifers. However, there are discrepancies between conservation guidelines (included in Natura 2000) and historical and palaeoecological data with regard to the distribution of conifers. Our aim was to bring new evidence to the debate on the conservation of conifers versus broadleaved trees at midelevations in Central Europe. We created a vegetation and land‐cover model based on pollen data for a highland area of 11,300 km² in the Czech Republic and assessed tree species composition in the forests before the onset of modern forestry based on 18th‐century archival sources. Conifers dominated the study region throughout the entire Holocene (approximately 40–60% of the area). Broadleaved trees were present in a much smaller area than envisaged by current ideas of natural vegetation. Rather than casting doubt on the principles of Central European nature conservation in general, our results highlight the necessity of detailed regional investigations and the importance of historical data in challenging established notions on the natural distribution of tree species.     

Composition and effects of inhalable size fractions of atmospheric aerosols in the polluted atmosphere: Part I. PAHs,PCBs and OCPs and the matrix chemical composition

Atmospheric particulate matter (PM) abundance, mass size distribution (MSD) and chemical composition are parameters relevant for human health effects. The MSD and phase state of semivolatile organic pollutants were determined at various polluted sites in addition to the PM composition and MSD. The distribution pattern of pollutants varied from side to side in correspondence to main particle sources and PM composition. Levels of particle-associated polycyclic aromatic hydrocarbons (PAHs) were 1–30 ng m^?3 (corresponding to 15–35 % of the total, i.e., gas and particulate phase concentrations), of polychlorinated biphenyls (PCBs) were 2–11 pg m^?3 (4–26 % of the total) and of DDT compounds were 2–12 pg m^?3 (4–23 % of the total). The PM associated amounts of other organochlorine pesticides were too low for quantification. The organics were preferentially found associated with particles <0.45 μm of aerodynamic equivalent diameter. The mass fractions associated with sub-micrometer particles (PM_0.95) were 73–90 %, 34–71 % and 36–81 % for PAHs, PCBs and DDT compounds, respectively. The finest particles fraction had the highest aerosol surface concentration (6.3–29.7)_×10^?6 cm^?1 (44–70 % of the surface concentration of all size fractions). The data set was used to test gas-particle partitioning models for semivolatile organics for the first time in terms of the organics' MSD and size-dependent PM composition. The results of this study prove that at the various sites particles with diverse size, matrix composition, amount of contaminants and toxicological effects occur. Legislative regulation based on gravimetric determination of PM mass can clearly be insufficient for assessment.     

In situ cadmium reclamation by freshwater bivalve Lamellidens marginalis from an industrial pollutant-fed river canal

The biofilter potential of the freshwater bivalve, Lamellidens marginalis was examined in cage experiments conducted in a river canal (Ichhapore, 24-Parganas, West Bengal, India) receiving industrial effluents from steel and metal factories as well as from an ordinance factory. Cadmium is one of the major contaminants in this river canal. Lamellidens collected from pollution free natural ponds, were sorted into three size groups (large: 59+/-3.2 g, 10+/-2.3 cm; medium: 30+/-2 g, 6+/-1.7 cm and small: 13+/-1.5 g, 4+/-1.2 cm) were held in cages at three different sites along a cadmium concentration gradient. Concentrations of cadmium were measured from water, sediment and different tissues of Lamellidens at weekly intervals using atomic absorption spectrophotometric methods. Cadmium uptake by Lamellidens in all media were highly concentration dependent in both summer and winter months. For all three size groups, cadmium uptake was maximum in the gills at the beginning of experiment, and liver at the later phase. Cadmium uptake was maximum in the small bivalves and minimum in the large bivalves groups. Cadmium uptake was 11-67% higher during summer than during the monsoon season for all tissues and size groups. Estimation of concentration factor revealed that tissues were saturated with cadmium during the 13-14th week after Lamellidens introduction during summer, but remained unsaturated during the monsoon season. It is concluded that Lamellidens might be considered as an efficient biofilter for reclamation of aquatic environment having sub-lethal concentrations of cadmium.     

Multivariate cluster analysis as a versatile tool for the quality assessment of short chain chloroparaffin quantification in environmental samples

The compositions of commercially available solutions of short chain chlorinated paraffins (SCCP) and technical mixtures were determined by high resolution gas chromatography (HRGC) electron capture negative ionisation (ECNI) combined with low resolution mass spectrometry (LRMS). Differences and similarities in the relative distribution of congeners and homologues were investigated by cluster analysis. Samples could be separated into two groups/clusters with similar molecular mass, chlorination degree and congener pattern. With the exception of one sample, the use of CH4-ECNI-MS led to an overestimation of the chlorine content. Moreover, the influence of different SCCP mixtures on the quantification was studied. The results showed that small differences in the chlorine content could sometimes cause substantial systematic errors of up to 119% despite similar homologue and congener patterns. Errors of quantification corresponded to the deviations between response factors of sample and standard. As a consequence SCCPs should be quantified with a standard matching the overall response factor of the sample.     

PCDDs, PCDFs and dioxin-like PCBs in food of animal origin (Slovakia)

The levels of 17 toxic polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and 12 non-ortho and mono-ortho polychlorinated biphenyls (PCBs) in 73 foodstuff samples of animal origin collected from shops and small farms in selected districts of Slovak Republic are presented in this paper. The concentrations expressed as WHO-TEQ in analysed samples ranged for PCDDs/PCDFs from 0.25 pg/g fat in pork to 75 pg/g fat in cod liver. The TEQ concentrations of non-ortho PCBs were between 0.007 and 181 pg/g fat and mono-ortho PCBs between 0.0083 and 66.5 pg/g fat. The mean concentrations in freshwater fish and imported species of marine fish were 0.089 pg TEQ/g fresh weight for PCDDs/PCDFs, 0.17 pg TEQ/g fresh weight for non-ortho PCBs and 0.034 pg TEQ/g fresh weight for mono-ortho PCBs. The mean total concentration of PCDDs/PCDFs and dioxin-like PCBs in samples of infant milk formula imported from EU countries was 0.98 pg TEQ/g fat.     

A worldwide survey of polychlorinated dibenzo-p-dioxins, dibenzofurans, and related contaminants in butter

The main source of human exposure to persistent organic pollutants (POPs) is, in general, food. In this study, 64 butter samples from 37 countries were analyzed to assess the global contamination of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), biphenyls (PCBs), hexachlorobenzene (HCB), and 2,2-bis (4-chlorophenyl)-1,1,1-trichloroethane (DDT) together with its major metabolites. The objectives of the study were to assess the presence of major organohalogen contaminants in butter, to trace geographical differences, and to determine toxic equivalents (TEQs) of PCDDs/Fs and dioxin-like PCBs in butter. The highest PCDD/F concentrations were found in butter from Korea with an average of 1.4 pg TEQ g(-1) lipid weight (l.w.). from PCDD/F and an additional contribution from the non- and mono-ortho-PCBs of 0.55 pg TEQ g(-1) l.w. Belgian butter showed average levels of 0.53 and 1.2 pg TEQ g(-1) l.w. for PCDDs/Fs and PCBs, respectively, but one sample of Belgium butter had a total TEQ level as high as 4.0 pg TEQ g(-1) l.w. Three out of five butter samples from Portugal showed similarly high PCDD/F TEQ levels. The sigmaPCB levels in European butter appeared to be somewhat higher than in the samples from the rest of the world. The average contribution of CB-153 to the total PCB concentration was 22% (SD 6.4, coefficient of variation 29%). Generally, the PCBs contributed around 60% of the total TEQ value, with CB-126 contributing approximately half of this value. This shows the important TEQ contribution from dioxinlike PCBs to the total TEQs. The highest HCB levels were found in butter samples from Russia, Ukraine, Belgium, and Slovenia. Low levels of HCB in butter were generally found in the Southern Hemisphere. Butter samples from countries from Eastern Europe had elevated sigmaDDT concentrations, with a particularly high concentration in Ukraine butter, followed by some Russian samples, Brazil, and the U.S.