罗红霉素短期冲击对活性污泥中氨氧化微生物丰度和多样性的影响  

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

作　　者：高景峰[1] 孙丽欣[1] 樊晓燕[1] 潘凯玲[1] 李定昌 

机构地区：[1]北京工业大学环境与能源工程学院城镇污水深度处理与资源化利用技术国家工程实验室,北京100124

出　　处：《环境科学》2017年第7期2961-2971,共11页Environmental Science

基　　金：国家自然科学基金项目(51378027;51578015);北京市自然科学基金项目(8162010)

摘　　要：本研究基于amo A基因,结合实时荧光定量PCR(quantitative real-time PCR,q PCR)和高通量测序技术研究罗红霉素(roxithromycin,ROX)短期冲击对活性污泥中氨氧化古菌(ammonia-oxidizing archaea,AOA)和氨氧化细菌(ammonia-oxidizing bacteria,AOB)丰度和多样性的影响.本研究共设置10种ROX浓度,不同浓度的ROX对氨氧化作用的影响差异明显,环境浓度(0.3~30μg·L^(-1))与中等浓度(300μg·L^(-1)和3000μg·L^(-1))的ROX并未对氨氧化作用产生影响;较高浓度(5 000~12 000μg·L^(-1))的ROX对氨氧化作用产生明显的抑制作用.环境浓度和中等浓度的ROX刺激了AOA增长,而较高浓度的ROX导致AOA的丰度下降.此外,除了环境中的痕量浓度(0.3μg·L^(-1)),其余浓度的ROX均导致AOB丰度下降,且下降趋势比AOA显著,说明AOA对ROX的耐受性高于AOB.高通量测序结果表明,在ROX的选择压下,AOA的OTUs多样性减少,AOB的OTUs多样性增加;但3个样品中最主要的AOA Candidatus Nitrososphaera gargensis的相对丰度随ROX浓度增加而增多,最主要的AOB Nitrosomonas eutropha的相对丰度随ROX浓度增加而减少,这同样说明了AOA对ROX的耐受性高于AOB.冗余分析结果表明:AOA Ca.Nitrososphaera gargensis、Candidatus Nitrosoarchaeum koreensis和AOB Nitrosomonas oligotropha、Nitrosomonas watsonii、Nitrosomonas halophilus均与ROX浓度呈正相关.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 amoA 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-1） and medium（300 μg·L-1and 3000 μg·L-1） levels of ROX did not affect ammonia oxidation, while the higher concentration（5000-12000 μg·L-1） 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-1） 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 Candidatus Nitrososphaera gargensis and Candidatus Nitrosoarchaeum koreensis. The insensitive AOB were Nitrosomonas oligotropha, Nitrosospira multiformis, Nitrosomonas watsonii and Nitrosomonas halophilus, 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 Ca. Nitrososphaera gargensis, Ca. Nitrosoarchaeum koreensis and AOB N. oligotropha, N. watsonii, N. halophilus were positively correlat

关 键 词：罗红霉素 活性污泥 氨氧化古菌 氨氧化细菌 高通量测序 

分 类 号：X172[环境科学与工程—环境科学] X703.1