Characterization of the genes involved in nitrogen cycling in wastewater treatment plants using DNA microarray and most probable number-PCR

To improve nitrogen removal performance of wastewater treatment plants (WWTPs), it is essential to understand the behavior of nitrogen cycling communities, which comprise various microorganisms. This study characterized the quantity and diversity of nitrogen cycling genes in various processes of municipal WWTPs by employing two molecular-based methods:most probable number-polymerase chain reaction (MPN-PCR) and DNA microarray. MPN-PCR analysis revealed that gene quantities were not statistically different among processes, suggesting that conventional activated sludge processes (CAS) are similar to nitrogen removal processes in their ability to retain an adequate population of nitrogen cycling microorganisms. Furthermore, most processes in the WWTPs that were researched shared a pattern:the nirS and the bacterial amoA genes were more abundant than the nirK and archaeal amoA genes, respectively. DNA microarray analysis revealed that several kinds of nitrification and denitrification genes were detected in both CAS and anaerobic-oxic processes (AO), whereas limited genes were detected in nitrogen removal processes. Results of this study suggest that CAS maintains a diverse community of nitrogen cycling microorganisms; moreover, the microbial communities in nitrogen removal processes may be specific.

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壬基酚、五氯酚及硝基苯对端足类河蜾蠃蜚(Corophium acherusicum)的毒性效应

为检测海洋中环境激素及芳烃类化合物对端足类生物的污染危害,实验选择端足类河蜾蠃蜚(Corophium acherusicum)为受试生物,研究了其在壬基酚、五氯酚、硝基苯三种有机污染物暴露下的96 h急性致死毒性效应和7 d慢性DNA损伤毒性效应。计算获得壬基酚、五氯酚和硝基苯对河蜾蠃蜚的96 h半致死浓度(LC50)分别为70、465、25 000μg·L-1,三种有机污染物对河蜾蠃蜚的毒性强弱顺序为壬基酚五氯酚硝基苯。运用碱解旋法检测壬基酚、五氯酚和硝基苯对河蜾蠃蜚DNA损伤的程度,计算得到7 d半效应浓度(EC50)分别为30、256、11 000μg·L-1。实验结果表明:三种有机污染物浓度的不断加大,引起河蜾蠃蜚DNA损伤程度的不断增加,呈显著的剂量-效应关系。     

Model reactions of chromium compounds with mammalian and bacterial cells†

Chromate uptake, reduction, cytotoxicity and mutagenicity were studied with human red blood cells, Chinese hamster ovary (CHO) cells and/or Salmonella typhimurium mutant cells. All cell types rapidly took up chromates whereas chromium(III) salts were excluded under the experimental conditions. Red blood cells reduced and accumulated chromium from chromate. At concentrations above 0.1 mM, chromate inactivated the red cell chromate carrier. Chromate above 0.01 mM inhibited CHO cell proliferation irrespective of the cations present. Chromate and two chromium(III) complexes were mutagenic with Salmonella mutants in the Ames’ assay. A model for chromate metabolism and genotoxicity is proposed.     

The cytotoxicity of nici2 for mammalian cells in culture†

The effects of NiCl₂ were studied in two human cell lines, HeLa and diploid embryonic fibroblasts as well as in V79 Chinese hamster cells and in L‐A mouse fibroblasts. NiCl₂ produces a dose‐dependent depression of proliferation, mitotic rate, and viability, accompanied by an increasing release of lactic dehydrogenase and stimulation of lactic acid production. The plating efficiency is reduced, as are DNA and protein synthesis and, to a lesser degree, RNA synthesis.

The cytotoxicity of NiCl₂ is comparable in degree to those of PbCl₂ and MnCl₂, but is weaker than those of HgCl₂ and CdCl₂. However, the different sensitivities of different cell lines must also be considered.

NiCl₂ effects are more severe in serum‐free medium than in medium containing serum or serum albumin indicating that serum constituents, notably albumin, bind the metal effectively and inhibit cellular uptake; this confirms earlier reports on the serum binding and slow uptake of NiCl₂.

Synchronized cells are most sensitive in the Gl and early S phases of the cell cycle. In the Painter test the depression of DNA synthesis persists following cessation of exposure to NiCI₂. These findings contribute an explanation for the known genotoxic effects of nickel.     

Biochemical properties of beryllium potentially relevant to its carcinogenicity†

The carcinogenicity of beryllium to several animal species is well established and evidence exists which strongly suggests that this is the case in human exposure. In this review several biochemical properties of the metallocarcinogen are considered including, the causation of cell transformation, and infidelity of DNA synthesis, inhibition of cell division and enzyme induction, and interference with regulatory mechanisms controlling gene expression. These effects are discussed in relation to beryllium chemistry, cellular accumulation mechanisms and distribution to subcellular organdies and molecular targets. It is suggested that the ultimate location and interactions of the metal ion in cell nuclei and its selective inhibition of certain protein phosphorylation reactions in particular are the biochemical effects potentially most relevant to induction of beryllium carcinogenesis.     

Uptake and genotoxicity of micromolar concentrations of cobalt chloride in mammalian cells

Literature data concerning the genotoxicity of cobalt salts have been conflicting. To establish appropriate incubation conditions, we conducted a series of uptake studies, before genotoxicity was determined by DNA strand break induction in HeLa cells and mutagenicity in V79 Chinese hamster cells. Co(II) is taken up by HeLa cells in a concentration‐dependent manner and is accumulated inside the cell. The uptake is preceded by a fast association step to the outer membrane, with no saturation up to 24 h. DNA strand breaks as determined by nucleoid sedimentation are induced at concentrations as low as 50μMCoCl₂. The induction is time‐dependent, showing the highest number of breaks after 4h incubation with no further increase up to 24h. CoCl₂ is mutagenic at the HPRT‐locus, enhancing the spontaneous mutation frequency 4.2‐fold at 100μ?. Besides direct interactions with DNA, the mutagenicity of CoCl₂ could also be due to a decrease in the Fidelity of DNA polymerisation.     

Biochemical speciation in chromium genotoxicity

The biochemical speciation of chromium compounds in mammalian cells is discussed with respect to uptake, metabolism, DNA binding and damaging. Whereas soluble hexavalent chromium is taken up rapidly and accumulated intracellularly after its reduction, compounds of trivalent chromium penetrate biomembranes about three orders of magnitude slower. Cr(VI) after its uptake is metabolised by electron donating compounds via Cr(V) to Cr(III) compounds. Chromium from various Cr(III) compounds, but not chromate, binds to chromatin in isolated cell nuclei. The DNA‐protein crosslinks and DNA strand breaks observed in rat liver and kidney after chromate administration are also found in vitro, when Cr(III) compounds (but not chromate) interacts with isolated nuclei. In the Chinese Hamster cell HGPRT mutation assay, three out of four tested Cr(III) complexes were found to be mutagenic. In a direct DNA strand break assay with supercoiled bacteriophage PM 2 DNA, neither chromate nor the four Cr(III) compounds tested caused nicks. However, the combined action of chromate plus glutathione as well as the isolated complex of pentavalent chromium, Na₄Cr(glutathione)₄, did cause DNA breaks. Reactive oxygen species are inferred to be the ultimate DNA nicking agents in this assay. In conclusion there appear to be two mechanisms of chromate genotoxicity; one with direct DNA damage caused by Cr(V) species and one via DNA‐protein crosslinks formed with Cr(III), the final reduction state of chromate.     

彗星实验检测渤海主要入海河流遗传毒性

利用彗星实验检测渤海区主要入海河流遗传毒性.以虾虎鱼为受试生物,暂养在河口水样中,染毒48h,取外周血细胞,运用彗星实验检测外周血细胞内DNA损伤程度,以尾相(TM)作为DNA损伤程度指标,并据此评估入海河流邻近海域遗传毒性风险.实验结果表明,入海河流中的特征污染物可导致虾虎鱼外周血细胞的DNA损伤,且损伤程度可以通过彗星实验定量分析,同时该试验方法操作简便、快速、灵敏度高,能够反映出多种污染因子的综合致毒能力.因此,通过彗星实验建立实验室检测入海河流遗传毒性方法具有可行行和创新性.     

威海荣成鹿角菜野生种群遗传多样性及资源保护策略

利用随机扩增多态性DNA（RAPD）方法对威海荣成地区鹿角菜（Silvetiasiliquosa）野生种群共30个个体的遗传多样性水平及遗传结构进行研究。结果表明,利用21条随机引物共检测到112个多态性位点,多态性位点百分率为93．75％,Nei基因多样性指数为0．3515±0．1352,Shannon多样性指数为0．5046±0．1126。通过聚类分析而划为一类的3个亚种群间的遗传分化指数为0．1349～0．2189,基因流为1．7793。研究结果表明威海荣成地区的鹿角菜种群遗传多样性较高,种群内存在较为明显的遗传分化,种群遗传资源不受遗传漂变的影响。针对当地鹿角菜遗传资源的现状,应加强现存种群的就地保护,恢复种群规模;进行取样养殖保护时则需避免因近交而造成种群资源退化。