[五氯酚对好氧颗粒污泥和活性污泥中氨氧化细菌的实时PCR定量分析]。

Guang-wei Li, He Liu, Feng Zhang, G. Du, Jian Chen
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引用次数: 3

摘要

利用特异引物直接从环境样品中扩增氨氧化菌(AOB) 16S核糖体DNA V2区。将纯化的PCR产物克隆到t载体上,经测序和Real-time PCR鉴定为AOB的16S rDNA片段。将重组质粒作为实时荧光定量PCR的标准分子样品进行AOB定量。采用Real-time PCR技术对受PCP影响的好氧颗粒污泥和活性污泥样品中AOB的数量进行了监测。结果表明,反应器中无PCP的好氧颗粒污泥和活性污泥中AOB的数量分别为4.28 × 10(7) 5.44 × 10(6)个细胞/g干燥污泥和2.51 × 10(9) +/- 8.61 × 10(8)个细胞/g干燥污泥。随着PCP浓度的增加(从0mg/L增加到50mg/L),两种污泥中AOB的数量无明显变化(P > 0.05)。AOB数量与氨氮去除率无显著相关(P > 0.05)。PCP对两种污泥中AOB的主要作用是抑制其代谢活性。
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[Real time PCR quantification of ammonia-oxidizing bacteria in aerobic granular sludge and activated sludge influenced by pentachlorophenol].
The V2 region of the 16S ribosomal DNA of the ammonia-oxidizing bacteria (AOB) was amplified directly from the environmental sample by using the specific PCR primers. The purified PCR product was cloned into T-vector and was identified as 16S rDNA fragment of AOB by sequencing and Real-time PCR method. Then, the recombined plasmid was used as standard molecule sample in Real-time PCR for AOB quantification. The numbers of the AOB were monitored in samples of both aerobic granular sludge and activated sludge influenced by PCP by using Real-time PCR. The results showed that the numbers of AOB in aerobic granular sludge and activated sludge were 4.28 x 10(7) 5.44 x 10(6) cells/g dried sludge and 2.51 x 10(9) +/- 8.61 x 10(8) cells/g dried sludge without PCP in the reactors, respectively. With the increase of PCP concentration (from 0mg/L to 50mg/L), the numbers of AOB in both types of sludge had no obvious change( P > 0.05) . The numbers of AOB had no obvious correlation with ammonia removal ( P > 0.05) . The main effect of PCP on AOB in both types of sludge was to inhibit their metabolic activity.
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