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罗红霉素对活性污泥中氨氧化微生物丰度和多样性的短期影响

[Short-term Effect of Roxithromycin on Abundance and Diversity of Ammonia-Oxidizing Microorganisms in Activated Sludge].

作者信息

Gao Jing-Feng, Sun Li-Xin, Fan Xiao-Yan, Pan Kai-Ling, Li Ding-Chang

机构信息

National Engineering Laboratory for Advanced Municipal Wastewater Treatment and Reuse Technology, College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124, China.

出版信息

Huan Jing Ke Xue. 2017 Jul 8;38(7):2961-2971. doi: 10.13227/j.hjkx.201612262.

Abstract

In this study, the short-term effect of roxithromycin(ROX) on the abundance and diversity of ammonia-oxidizing archaea(AOA) and ammonia-oxidizing bacteria(AOB) based on gene in activated sludge were investigated by high-throughput sequencing and quantitative real-time PCR(qPCR). High-throughput sequencing overcomes the drawbacks of low sequencing depth, significant randomness and great bias of traditional Sanger sequencing. This approach can provide enough sequencing depth to comprehensively investigate the sensitive and insensitive ammonia-oxidizing microorganisms under ROX selective pressure. Lab-scale reactors were operated under ten different ROX levels. The results indicated that the environmental(0.3-30 μg·L) and medium(300 μg·Land 3000 μg·L) levels of ROX did not affect ammonia oxidation, while the higher concentration(5000-12000 μg·L) of ROX showed a significant negative effect on ammonia oxidation. The environmental and medium levels of ROX stimulated the growth of AOA, however, the higher level of ROX decreased the abundance of AOA. In addition, different levels of ROX(except 0.3 μg·L) caused the decrease of the abundance of AOB, which suggested that AOA was less sensitive than AOB under ROX selective pressure. The results of high-throughput sequencing showed that ROX selective pressure caused the decrease of the numbers of OTUs for AOA and increase of that for AOB. The insensitive AOA, accounting for 57.70%-97.81% of the total sequences, were Nitrososphaera gargensis and Nitrosoarchaeum koreensis. The insensitive AOB were and , accounting for 0.76%-5.10% of the total sequences. These results also indicated that AOA was insensitive to ROX, but AOB was sensitive to ROX. RDA analyses showed that AOA . Nitrososphaera gargensis, . Nitrosoarchaeum koreensis and AOB were positively correlated with ROX concentrations.

摘要

在本研究中,通过高通量测序和定量实时聚合酶链反应(qPCR),研究了罗红霉素(ROX)对活性污泥中基于基因的氨氧化古菌(AOA)和氨氧化细菌(AOB)丰度及多样性的短期影响。高通量测序克服了传统桑格测序测序深度低、随机性大及偏差大的缺点。该方法能够提供足够的测序深度,以全面研究在ROX选择压力下敏感和不敏感的氨氧化微生物。实验室规模的反应器在十种不同的ROX水平下运行。结果表明,环境水平(0.3 - 30 μg·L)和中等水平(300 μg·L和3000 μg·L)的ROX不影响氨氧化,而较高浓度(5000 - 12000 μg·L)的ROX对氨氧化有显著负面影响。环境和中等水平的ROX刺激了AOA的生长,然而,较高水平的ROX降低了AOA的丰度。此外,不同水平的ROX(除0.3 μg·L外)导致AOB丰度下降,这表明在ROX选择压力下AOA比AOB不敏感。高通量测序结果表明,ROX选择压力导致AOA的操作分类单元(OTU)数量减少,而AOB的OTU数量增加。不敏感的AOA,占总序列的57.70% - 97.81%,为加尔各答亚硝化球菌(Nitrososphaera gargensis)和韩国亚硝化古菌(Nitrosoarchaeum koreensis)。不敏感的AOB为 和 ,占总序列的0.76% - 5.10%。这些结果还表明AOA对ROX不敏感,但AOB对ROX敏感。冗余分析(RDA)表明,AOA、加尔各答亚硝化球菌、韩国亚硝化古菌和AOB与ROX浓度呈正相关。

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