Induction of unscheduled DNA synthesis and micronucleus formation in Syrian hamster embryo fibroblasts treated with cysteine S-conjugates of chlorinated hydrocarbons

S-(chloroethyl)-cysteine (CEC) and S-(1,2-dichlorovinyl)cysteine (DCVO) have been proposed as intermediates in the metabolic transformation of the carcinogens 1,2-dichloroethane and 1,1,2-trichloroethylene. We have tested the ability of CEC and DCVC to induce DNA repair and genotoxic effects at the chromosomal level by comparative assessment of unscheduled DNA synthesis induction and micronucleus formation in Syrian hamster embryo fibroblasts. CEC induced a potent and dose-dependent response in both assays, whereas DCVC treatment resulted in a comparatively weak induction of DNA repair and failed to raise micronucleus formation above control rates. Inhibition of cysteine conjugate \gB-lyase diminished the effect of DCVC, but had no influence on the genotoxicity of CEC either in the unscheduled DNA synthesis or micronucleus assay.Abbreviations AOAA aminooxyacetic acid - CEC S-(chloroethyl)-cysteine; \gB-lyase, cysteine conjugate -lyase - DCE 1,2-dichloroethane - DCVC S(1,2-dichlorovinyl)-cysteine - GSH glutathione - HU hydroxyurea - IBR IBR-modified Dulbecco's Eagle's reinforced medium - MN2 micronuclei/2,000 cells - 4-NQO 4-nitroquinoline-1-oxide - SHE Syrian hamster embryo fibroblasts; ³H-Thd, ³H-thymidine - TCE 1,1,2-trichloroethylene - UDS unscheduled DNA synthesis     

一株脱卤单胞菌属有机卤呼吸细菌的分离纯化与基础特征

【目的】分离鉴定新型有机卤呼吸细菌,拓展有机氯污染物降解菌种资源。【方法】在限制性培养基中,基于特定脱卤微生物的能量代谢特点以及对抗生素的抗性特征,应用绝迹稀释法从脱氯富集培养物中分离纯化新型有机卤呼吸细菌。通过添加酵母提取物和聚合酶链反应-限制性片段长度多态性等方法鉴定菌株纯度。通过考察细胞形态、16S rRNA系统发育树以及对有机氯化物的利用能力来描述此新型菌株的基本特征。【结果】菌株GP是一株来自脱卤单胞菌属(Dehalo genimonas)的新型有机卤呼吸细菌,可耐受1.0 g/L的氨苄青霉素和0.1 g/L的万古霉素,其细胞平面形态为直径0.4–0.8μm的不规则圆形。在乙酸作碳源、氢气作电子供体的条件下,菌株GP可利用1,1-二氯乙烯和一氯乙烯作为电子受体进行有机卤呼吸,脱氯终产物为无毒害的乙烯。16SrRNA系统发育分析表明,菌株GP与Dehalo genimonas formicexedens菌株NSZ-14有高达99.5%的同源性,但两菌株对有机氯化物底物的利用范围不同。【结论】从脱氯富集培养物中分离纯化出一株新型有机卤呼吸细菌,为深入研究脱卤单胞菌这一重要脱卤微生物的遗传学信息和生理生化性质提供了新材料。     

Inhibitory effect of chlorinated guaiacols on methanogenic activity of anaerobic digester sludge

Of four chlorinated guaiacols, tetrachloroguaiacol at 62 M inhibited acetate methanogenesis, the strongest decreasing activity by 50%. 4,5,6-Trichloroguaiacol, 4,5-dichloroguaiacol, and 4-chloroguaiacol showed 50% inhibition at 0.13, 0.32, and 1.50 mM, respectively. Degradation test results of volatile fatty acids (acetic, propionic, and butyric acid) by anaerobic digester sludge (stored 5 weeks) indicated that syntrophic butyrate degraders of this sludge were more sensitive to tetrachloroguaiacol than acetoclastic methanogens and syntrophic propionate degraders.     

The role of microbial reductive dechlorination of TCE at a phytoremediation site

In April 1996, a phytoremediation field demonstration site at the Naval Air Station, Fort Worth, Texas, was developed to remediate shallow oxic ground water (< 3.7 m deep) contaminated with chlorinated ethenes. Microbial populations were sampled in February and June 1998. The populations under the newly planted cottonwood trees had not yet matured to an anaerobic community that could dechlorinate trichloroethene (TCE) to cis-1,2-dichloroethene (DCE); however, the microbial population under a mature (approximately 22-year-old) cottonwood tree about 30 m southwest of the plantings had a mature anaerobic population capable of dechlorinating TCE to DCE, and DCE to vinyl chloride (VC). Oxygen-free sediment incubations with contaminated groundwater also demonstrated that resident microorganisms were capable of the dechlorination of TCE to DCE. This suggests that a sufficient amount of organic material is present for microbial dechlorination in aquifer microniches where dissolved O2 concentrations are low. Phenol, benzoic acid, acetic acid, and a cyclic hydrocarbon, compounds consistent with the degradation of root exudates and complex aromatic compounds, were identified by gas chromatography/mass spectrometry (GC/MS) in sediment samples under the mature cottonwood tree. Elsewhere at the site, transpiration and degradation by the cottonwood trees appears to be responsible for loss of chlorinated ethenes.     

Ethene as an auxiliary substrate for the cooxidation of cis-1,2-dichloroethene and vinyl chloride

Cultures able to dechlorinate cis-1,2-dichloroethene (cDCE) were selected with ethene (3–20%, v/v) as the sole source of carbon and energy. One mixed culture (K20) could degrade cDCE (400 μmol l^–1) or vinyl chloride (100 μmol l^–1) in the presence of ethene (≤ 80 μmol l^–1 and ≤ 210 μmol l^–1, respectively). This culture consists of at least five bacterial strains. All five strains were able to degrade cDCE cometabolically in pure culture. The mixed culture K20 was highly tolerant against cDCE (up to 6 mmol l^–1 in the liquid phase). Degradation of cDCE (200 μmol l^–1) was not affected by the presence of trichloroethene (100 μmol l^–1) or tetrachloroethene (100 μmol l^–1). Transformation yields (T_y, defined as unit mass of chloroethene degraded per unit mass of ethene consumed) of the mixed culture K20 were relatively high (0.51 and 0.61 for cDCE and vinyl chloride, respectively). The yield for cDCE with ethene as auxiliary substrate was ninefold higher than any values reported with methane or methane/formate as auxiliary substrate. The viability of the cells of the mixed culture K20 (0.3 mg of cells ml^–1) was unaffected by the transformation of ≤ 200 μmol l^–1 cDCE in 300 min. Received: 9 March 1999 / Accepted: 21 July 1999     

Histological assessment of selected tissues in tigerfish (Hydrocynus vittatus Castelnau, 1861) from the Sanyati Basin,Lake Kariba,Zimbabwe: a DDT-sprayed area

A pilot investigation was conducted in 2014 in the Sanyati Basin of Lake Kariba to ascertain whether long-term DDT spraying in the Kariba catchment had a negative effect on fish health. The aim was to assess the health of tigerfish, Hydrocynus vittatus, by means of histological analysis and to analyse water, sediment and tissue samples for bioaccumulated levels of chlorinated pesticides, including DDT. Eighteen tigerfish were collected by seine netting along the north-eastern shoreline of the Sanyati Basin in April 2014 and samples of their gill, liver, kidney, muscle and brain tissue were processed for histology and assessed using light microscopy. No detectable levels of DDT and/or its metabolites were found in the water or sediment samples and only a low concentration of p,p′-DDE in fish tissues. No major histological alterations were observed in the fish tissues. Consequently, there seems to be no risk of DDT exposure following the consumption of tigerfish from the Sanyati Basin of Lake Kariba. These results were unexpected, given the historical use of DDT within the current study area. It is recommended that these findings be compared with those from other regions of Lake Kariba.     

Field Note: Phytoremediation of Chlorinated Ethenes in Seepline Sediments: Tree Selection

Phytoremediation of chlorinated ethene (CE)-contaminated water was investigated at the Savannah River Site in Aiken, SC, USA. Perchloroethylene (PCE) and trichloroethylene (TCE) are present where CE-contaminated groundwater currently outcrops in seepline soils. Results of constructed and planted test cells, filled with soil from a noncontaminated seepline area and supplied with CE-contaminated groundwater (48 ppb) in the field for one season are presented. These test cells were planted with loblolly pines, hybrid poplars, coyote willow, and sweet gum. Cis-dichloroethylene (cDCE), a byproduct from rhizosphere microbial activity, was detected in the soils as well as some tree tissues. All trees tested were found to uptake both PCE and TCE (5–50 pbb/gm dry wt).