Evaluation of bendiocarb cytotoxicity in mammalian and insect cell cultures

There is an increasing need for rapid and easily interpreted in vitro assays to screen for possible cytotoxicity of pesticides. The objective of this study was to investigate the effect of the carbamate insecticide bendiocarb on mammalian and insect cell cultures. The cytotoxicity of this insecticide was evaluated by cell proliferation and cellular damage was assessed by evaluation of the cytopathic effect and lactate dehydrogenase (LDH) leakage. Cells of insect origin (Sf21) were the most sensitive to bendiocarb with significant (P < 0.01) suppression of their proliferative activity ranging from 10(-1)-10(-5) M. However, significant suppression of proliferative activity was also recorded in rat liver cells (WBF344; 10(-1)-10(-3) M; P < 0.01-0.05) and rabbit kidney cells (RK13; 10(-1) M; P < 0.01). In contrast with the proliferation activity of cells, a cytopathic effect based on cellular damage and LDH leakage into the medium was observed only at the highest concentration (10(-1) M) in RK 13 and WBF344 cells, but not in the Sf21 insect cell line. Our results indicate that bendiocarb exposure caused a cell-type dependent decrease in cell proliferation; however, cell damage and LDH leakage into the medium were not present or were strongly limited, dependent on the cell phenotype. Cell proliferation was shown as a sensitive indicator for evaluation of the cytotoxic effect of bendiocarb in vitro; on the other hand, microscopic signs of cellular damage and LDH leakage were insufficient in vitro markers.     

Source identification of personal exposure to fine particulate matter using organic tracers

Personal exposure to fine particulate matter (PM2.5) is due to both indoor and outdoor sources. Contributions of sources to personal exposure can be quite different from those observed at ambient sampling locations. The primary goal of this study was to investigate the effectiveness of using trace organic speciation data to help identify sources influencing PM2.5 exposure concentrations. Sixty-four 24-h PM2.5 samples were obtained on seven different subjects in and around Boulder, CO. The exposure samples were analyzed for PM2.5 mass, elemental and organic carbon, organic tracer compounds, water-soluble metals, ammonia, and nitrate. This study is the first to measure a broad distribution of organic tracer compounds in PM2.5 personal samples. PM2.5 mass exposure concentrations averaged 8.4 μg m^?3. Organic carbon was the dominant constituent of the PM2.5 mass. Forty-four organic species and 19 water-soluble metals were quantifiable in more than half of the samples. Fifty-four organic species and 16 water-soluble metals had measurement signal-to-noise ratios larger than two after blank subtraction.The dataset was analyzed by Principal Component Analysis (PCA) to determine the factors that account for the greatest variance. Eight significant factors were identified; each factor was matched to its likely source based primarily on the marker species that loaded the factor. The results were consistent with the expectation that multiple marker species for the same source loaded the same factor. Meat cooking was an important source of variability. The factor that represents meat cooking was highly correlated with organic carbon concentrations (r = 0.84). The correlation between ambient PM2.5 and PM2.5 exposure was relatively weak (r = 0.15). Time participants spent performing various activities was generally not well correlated with PCA factor scores, likely because activity duration does not measure emissions intensity. The PCA results demonstrate that organic tracers can aid in identifying factors that influence personal exposures to PM2.5.     

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.     

Chloride concentration affects soil microbial community

We studied the effect of increased inorganic chloride concentration on forest soil microflora in a laboratory experiment. Microbial DNA extracted from experimental soil samples was amplified with PCR using primer pairs specifically amplifying bacterial, eukaryotic and fungal DNA fragments. The resulting amplified DNA was further used for terminal restriction fragment length polymorphism (TRFLP) analysis. Our work revealed that chloride concentration affects the indigenous microbial community in experimental soil. This was documented on an unidentified microorganism whose DNA was detectable in soil high in chloride but was not found in soil with low chloride concentration. The presence of the organism responsive to increased chloride concentration was associated with the highest observed value of chlorination of humic acid, suggesting possible role of this organism in soil chlorine turnover. High chloride concentration in the soil tended to decrease the rate of degradation of trichloroacetic acid. The problems connected with measurement of chlorination rates in soil are discussed.     

Uranium and neptunium retention mechanisms in Gallionella ferruginea/ferrihydrite systems for remediation purposes

The ubiquitous β-Proteobacterium Gallionella ferruginea is known as stalk-forming, microaerophilic iron(II) oxidizer, which rapidly produces iron oxyhydroxide precipitates. Uranium and neptunium sorption on the resulting intermixes of G. ferruginea cells, stalks, extracellular exudates, and precipitated iron oxyhydroxides (BIOS) was compared to sorption to abiotically formed iron oxides and oxyhydroxides. The results show a high sorption capacity of BIOS towards radionuclides at circumneutral pH values with an apparent bulk distribution coefficient (K_d) of 1.23 × 10⁴ L kg^?1 for uranium and 3.07 × 10⁵ L kg^?1 for neptunium. The spectroscopic approach by X-ray absorption spectroscopy (XAS) and ATR FT-IR spectroscopy, which was applied on BIOS samples, showed the formation of inner-sphere complexes. The structural data obtained at the uranium L_III-edge and the neptunium L_III-edge indicate the formation of bidentate edge-sharing surface complexes, which are known as the main sorption species on abiotic ferrihydrite. Since the rate of iron precipitation in G. ferruginea-dominated systems is 60 times faster than in abiotic systems, more ferrihydrite will be available for immobilization processes of heavy metals and radionuclides in contaminated environments and even in the far-field of high-level nuclear waste repositories.

     

Possible effects of persistent organochlorinated pollutants cocktail on thyroid hormone levels and pituitary-thyroid interrelations

In polluted district of Michalovce in East Slovakia (POLL) and two districts with background pollution (BCGR) 2046 adults (834 males and 1212 females aged 20-75 years) were examined. Serum levels of thyrotropin (TSH), free thyroxine (FT4), total triiodothyronine (TT3) and antithyroperoxidase antibodies (TPOab) were estimated by electrochemiluminiscent assay and also these of 15 polychlorinated biphenyl congeners (PCBs), p,p'-DDE, p,p'-DDT, hexachlorobenzene (HCB) and hexachlorocyclohexane were measured by high resolution gas chromatography/mass spectrometry. In addition, also dioxins, furans, coplanar- and mono-ortho-PCBs as well as selected hydroxylated and methylsulphonated PCBs and DDE metabolites were measured by appropriate methods based on gas chromatography/mass spectrometry principle. In POLL significantly higher levels of all organochlorines were found than these in BCGR. When pooled values from both areas were stratified in terms of PCBs level and treated as continuous variables, positive association of PCBs with FT4 and TT3 was found, the latter two being also mutually associated. However, within the category of PCBs level <530 ng/glipid (n=232) the association between PCBs and both the FT4 (p<0.09) and TT3 (p<0.03) was negative and any association of these was not found within the category of PCBs level of 531-1000 ng/g (n=691). In contrast, in the category of 531-2000 ng/g (n=1307) positive association appeared between PCBs and FT4 (p<0.001) as well as TT3 (p<0.05). Highly significant association of PCBs with FT4 (p<0.001) was further found in the categories with PCBs level of 1001-101414 ng/g (n=1307) and 2001-101414 (n=1123), while significant association with TT3 was observed only in the category of 531-2000 ng/g. Such findings suggest possible threshold level in positive effect of PCBs on FT4 and TT3 level which seems to be individual and located somewhere around the PCBs level of 1000 ng/g. However, highly significant negative association of both FT4 and TT3 with TSH was found in each of above indicated PCBs categories. Considerable difference in FT4 and TT3 level between large groups of subjects with the same range of PCBs level was also found suggesting different individual susceptibility to the effects of organochlorines. Among a total of 26 cases from POLL with very low TSH level (<0.5 mU l(-1)) 13 cases showed very high level of PCBs, FT4 and TT3, thus supporting a hypothesis on a novel sporadic form of high PCBs related peripheral subclinical hyperthyroidism possibly resulting from the long-term disruption of equilibrium between bound and free thyroxine in plasma by high PCBs level followed by a striking inhibition of TSH release from the pituitary.     

Brominated flame retardants and related chlorinated persistent organic pollutants in fish from river Elbe and its main tributary Vltava

Brominated flame retardants (BFRs) are widely used industrial chemicals, residues of which can be nowadays found in all environmental compartments. The widespread presence of BFRs in various environmental compartments and food chain is a consequence of both their broad application area and physico-chemical properties, such as resistance to degradation and high lipophilicity. Alike in the case of other halogenated persistent organic pollutants (POPs), fish can be used as a bioindicator of aquatic environment pollution. In presented study, conducted in the year 2005, altogether 80 samples representing the most abundant fresh water fish species, viz. chub (Leuciscus cephalus), bream (Abramis brama), and perch (Perca fluviatilis) collected in 11 sampling sites located at Elbe and Vltava (Moldau) rivers were examined for levels of major BFRs. Without any exception, BFRs were detected in all fish samples. BDE 47 was the dominating congener in all fish species. This fact was not surprising, since it used to be the main component in various kinds of technical mixtures. With regard to relatively high levels of BDE 47 in fish tissue, as compared to other BFRs, and considering strong correlation with the total PBDEs content, simplified laboratory examination and, consequently, increased samples throughput can be obtained when only this congener is monitored. The potential of comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GCxGC-TOFMS), to provide more comprehensive information on the bioaccumulating chemicals occurring in fish samples, has been demonstrated in this study.     

Distribution pattern of PCBs,HCB and PeCB using passive air and soil sampling in Estonia

Background, aim, and scope  

Passive air sampling survey of the Central and Eastern Europe was initiated in 2006. This paper presents data on toxic organic compounds such as polychlorinated biphenyls (PCB 28, 52, 101, 118, 153, 138, and 180), hexachlorobenzene (HCB), pentachlorobenzene (PeCB), hexachlorocyclohexane compounds (α-HCH, β-HCH,γ-HCH, δ-HCH), and dichloro-diphenyl-trichloroethane (DDT) compounds (p,p′DDE, p,p′DDD, p,p′DDT, o,p′DDE, o,p′DDD, and o,p′DDT) determined in ambient air and soil samples collected at Estonian monitoring stations.     

Screening of short- and medium-chain chlorinated paraffins in selected riverine sediments and sludge from the Czech Republic

Wide distribution of chlorinated paraffins in the environment has already been demonstrated in several studies; however, information about their levels in the Central Europe is still very limited. First study focused on the SCCP contamination of the Czech aquatic environment have been performed recently, and its results motivated the authors to analyze sediments from a wide set of the Czech rivers in order to obtain more detailed information. Thirty-six sediment samples from eleven rivers and five drainage vents neighboring the chemical factories were analyzed; special attention was paid to the industrial areas. For the first time in the Czech Republic, medium-chain in addition to short-chain chlorinated paraffins were analyzed using GC-ECNI-MS. Chlorinated paraffins were detected in sediment samples on the concentration levels up to 347 ngg(-1) for short-chain chlorinated paraffins, and 5575 ngg(-1) for medium-chain chlorinated paraffins. Average chlorination degree of SCCPs was 65%.