油田硫酸盐还原菌快速定量检测方法

现有油田废水中硫酸盐还原菌检测周期长、检测费用较高,本研究应用聚合酶链式反应(PCR)技术与倍比稀释法(MPN)相结合的DSR-MPN-PCR法,对硫酸盐还原菌进行快速定量检测.从废水中制备了直接用于PCR扩增的菌液,保证了定量准确性;建立以硫酸盐还原菌亚硫酸盐还原酶基因(Dsr)为靶位点的通用探针DSR1F和DSR5R的反应体系和扩增条件.结果表明,该方法检测灵敏度明显比液体稀释培养法高2个数量级,真实地表征了废水中实际的SRB菌数量,整个操作过程需要3～4　h,检测结果非常稳定,降低了检测费用,可以在生产中应用.     

Evaluation of mixture effects in a crude extract of compost using the CALUX bioassay and HPLC fractionation

Potential synergistic interactions between polycyclic aromatic hydrocarbons in a household sewage sludge compost extract were investigated using the Dioxin-Responsive Chemical-Activated Luciferase gene eXpression (DR-CALUX) assay and reverse-phase high-performance liquid chromatography (RP-HPLC) fractionation. The biological activity of the crude extract was measured in vitro using the CALUX assay. The CALUX activity of the extract was as potent as 360-pg CALUX-TEQ (2,3,7,8-TCDD equivalent value) per g sample, this was 70 times above the WHO-TEQ value which was derived from chemical analyses of dioxins/furans and dioxin-like PCBs of the mixture. The CALUX activity pattern of the crude extract and the retention times of 26 polycyclic aromatic compounds (PACs), as determined by RP-HPLC on an octadecylsilica column, suggested that the dioxin-like compounds with the log K(OW) (n-octanol/water partition coefficient) values corresponding to 6.0-7.0 contributed highly to the whole activity. The CALUX activity of the crude extract was three times the sum of the CALUX activities of the RP-HPLC separated fractions. Mixture effects were assessed by co-exposure of each HPLC fraction and 2,3,7,8-TCDD to the cells. The four concentration levels of added 2,3,7,8-TCDD corresponded to the TEQ value in the original compost sample. The experimental CALUX activity was higher than the predicted CALUX activity for some fractions. It was demonstrated that some compounds in the compost sample interacted synergistically with 2,3,7,8-TCDD in terms of dioxin-like activity. This finding points out the necessity for detailed investigation of synergistic effects in environmental samples.     

彗星试验的改良及其在工业废水监测中的应用

为了探讨彗星试验在工业废水监测中的应用,本文将常规的细胞暴露方式改良为浸泡染毒,既提高了实验的敏感性,又使得该试验直接用于监测工业废水的遗传毒性成为可能．作者用改良彗星试验直接检测了几种工业废水所引起的V79细胞DNA损伤．结果表明,所测的各种工业废水均含有DNA损伤剂,能够诱发培养的V79细胞DNA链断裂,其中以制革厂废水的DNA损伤作用最强．本研究显示出改良彗星试验在综合评价各种废水遗传毒性方面的广阔应用前景．     

长江南京段水体中有机污染物的遗传毒性研究

运用人外周血淋巴细胞彗星试验和蚕豆根尖细胞微核试验对长江南京段水体中有机污染物的遗传毒性进行了研究. 结果表明:长江南京段水体中的有机污染物对人外周血淋巴细胞和蚕豆根尖细胞均产生了不同程度的损伤,存在明显的遗传毒性,有机物是导致水体遗传毒性的主要因素. 试验结果与水体的有机污染状况基本一致. 彗星试验结果及趋势与微核试验相吻合,但前者更为敏感. 彗星试验和微核试验的结合使用在水环境的遗传毒性监测方面具有较大的应用价值.      

Function of arsATorf7orf8 of Bacillus sp. CDB3 in arsenic resistance

Bacillus sp. CDB3 isolated from an arsenic contaminated cattledip site possesses an uncommon arsenic resistance (ars) operon bearing eight genes in the order of arsRYCDATorf7orf8. We investigated the functions of arsA, arsT, orf7 and orf8 in arsenic resistance using a plasmid-based gene knockout approach in the ars genedeficient Escherichia coli strain AW3110. The CDB3 arsA genewas shown to play a significant role in resistance, suggesting that the encoded ArsA may couplewith the arsenite transporter, forming an ArsAY complex that can enhance arsenite extrusion efficiency. Thedisruption of either arsT or orf7was not observed to affect arsenic resistance in the heterologous E. coli host, but their involvement in arsenic resistance can not be excluded. The orf8 gene is predicted to encode a putativedual-specificity protein phosphatasewhich also shares certain homology to arsenate reductases. The function loss of orf8 resulted in a remarkabledecrease in resistance to arsenate, though not arsenite. To examine if this effectwasdue to the reduction of arsenate by orf8, the arsC genewithin the 8-gene operonwasdisrupted. The resulting abolishment of arsenate resistance suggests that the involvement of orf8 in arsenic resistance is not via reductase activity.     

太湖不同湖区蓝藻细胞裂解速率的空间差异

2009年在太湖蓝藻水华形成初期(五月)、盛发期(九月)和衰亡期(十月和十一月),运用基于颗粒态酯酶,溶解性酯酶以及酯酶衰变常数测定的酯酶活性方法对不同湖区(藻型和草型湖区)蓝藻的细胞裂解速率进行了计算,在测定颗粒态酯酶、溶解性酯酶活性时,同步分析了太湖优势种群中蓝藻叶绿素a的含量.统计分析结果表明,叶绿素a的浓度与颗粒态酯酶、溶解性酯酶活性有很好的相关性,说明以酯酶活性为指标来计算太湖蓝藻细胞裂解速率是可行的.对不同湖区的细胞裂解速率进行比较,可见湖心和西太湖在蓝藻水华形成初期细胞裂解速率分别为0.072,0.048d-1.水华盛发期以及水华衰亡期,湖心和西太湖的细胞裂解速率分别为0.074～0.770d-1,0.014～0.110d-1.太湖湖心磷酸盐浓度比西太湖低,所以蓝藻生长速率慢,导致细胞裂解速率比西太湖高.但是,在梅梁湾和贡湖,衰亡末期磷酸盐浓度比其它月份高,细胞裂解速率也高.4个采样点在衰亡末期的细胞裂解速率比水华形成初期,暴发期和衰亡初期要高,可能的原因是气温和水体温度下降导致蓝藻生长速度减慢.本研究结果表明,太湖蓝藻细胞裂解速率有明显的空间差异,其具体的影响因素很多,营养盐只是其中一个.     

太湖梅梁湾有机污染物对纤细裸藻（Euglena gracilis）的遗传毒性效应

采用GC-MS方法分析了太湖梅梁湾水样的主要有机污染物成分和含量,共检测出15种目标化合物,表明该区域水体已经受到有机物污染;纤细裸藻(Euglena gracilis)在低浓度浓缩水样处理下(1倍组)生长无显著影响,叶绿素a、b含量和类胡萝卜素含量比对照组显著增加,而高浓度下(5倍和10倍组)生长明显受到抑制,叶绿素a和类胡萝卜素含量呈下降趋势;抗氧化酶系统中SOD和POD活性呈上升趋势,显示有机污染物胁迫可诱导抗氧化酶活性;彗星试验中,Olive尾矩和尾动量增加,纤细裸藻细胞DNA损伤程度随着太湖水样污染物浓度增加而加重,呈现出明显的剂量-效应关系,提示太湖梅梁湾水样具有潜在致突变性.结果表明,慧星试验和SOD活性试验结合使用在水环境中的遗传毒性监测方面具有较大的应用价值,适合作为水体有机污染监测的生物标志物.     

Assessment of source water contamination by estrogenic disrupting compounds in China

Detection of estrogenic disrupting compounds (EDCs) in drinking waters around China has led to rising concerns about health risks associated with these compounds. There is, however, a paucity of studies on the occurrence and identification of the main compounds responsible for this pollution in the source waters. To fill this void, we screened estrogenic activities of 23 source water samples from six main river systems in China, using a recombinant two-hybrid yeast assay. All sample extracts induced significant estrogenic activity, with E2 equivalents (EEQ) of raw water ranging from 0.08 to 2.40 ng/L. Additionally, 16 samples were selected for chemical analysis by gas chromatography-mass spectrometry. The EDCs of most concern, including estrone (E1), 17βup-estradiol (E2), 17αup-ethinylestradiol (EE2), estriol (E3), diethylstilbestrol (DES), estradiol valerate (EV), 4-t-octylphenol (4-t-OP), 4-nonylphenols (4-NP) and bisphenol A (BPA), were determined at concentrations of up to 2.98, 1.07, 2.67, 4.37, 2.52, 1.96, 89.52, 280.19 and 710.65 ng/L, respectively. Causality analysis, involving comparison of EEQ values from yeast assay and chemical analysis identified E2, EE2 and 4-NP as the main responsible compounds, accounting for the whole estrogenic activities (39.74% to 96.68%). The proposed approach using both chemical analysis and yeast assay could be used for the identification and evaluation of EDCs in source waters of China.     

四膜虫彗星实验在环境水质检测中的应用

介绍四膜虫作为生物材料在彗星实验水质检测中的应用,探讨了将单细胞原生生物应用于彗星实验在环境水质检测、遗传毒性检测等方面的应用优势和发展前景。研究结果表明:过氧化氢浓度与细胞损伤之间呈现良好的剂量-效应关系,东湖水和长江水未造成四膜虫DNA的损伤,印染厂污水对细胞有严重的损伤作用,可能会影响正常的生态环境。     

In vitro toxic effects of heavy metals on fungal growth and phosphate-solubilising abilities of isolates obtained from Phragmites australis rhizosphere

In this research, we evaluated the toxic effect of metal ions on mycelial growth and phosphate-solubilising activity of soil-borne micromycetes isolated from the Phragmites australis rhizosphere using Pikovskaya-agar plates supplemented with four metal concentrations. The diameter growth rate (DGR) decreased as the metal concentration rise for all tested fungi. Trichoderma atroviride had the fastest growth rate (1.48?cm²?day^?1) and was the least susceptible to Al³⁺, Cd²⁺, Cr³⁺, Cu²⁺ and Pb²⁺ with a median effective concentration (MEC₅₀) of 12.19, 0.48, 4.51, 11.44 and 50.05?mM, respectively. Aspergillus japonicus was the most tolerant to Co²⁺, Ni²⁺ and Zn²⁺, with MEC₅₀ values of 3.36, 1.095 and 2.34?mM, respectively. Penicillium italicum was the most tolerant to Cr6⁺ (MEC₅₀?=?0.677?mM). The ability to solubilise phosphate remained, despite the decrease in the DGR, and P. italicum and Penicillium dipodomyicola had the highest Phosphate Solubilisation Indexes (PSIs) at 1.97 and 2.12, respectively. In particular, P. italicum recorded the highest PSI of all the studied isolates at 0.62?mM Cr³⁺ (PSI?=?4.74). A. japonicus and T. atroviride were the most tolerant isolates to all tested metals, which suggests that these isolates are promising candidates for further study with regard to mycoremediation and biofertilisation of metal-polluted soils.