处理水产养殖污水潜流湿地中的厌氧氨氧化菌群特征

厌氧氨氧化(anaerobic ammonium oxidation,ANAMMOX)是潜流湿地净化污水的重要功能,目前关于潜流湿地厌氧氨氧化特征还不清楚.为了解处理水产养殖污水潜流湿地中的厌氧氨氧化特点,采用16S rRNA基因克隆文库和实时荧光定量PCR技术以及多样性分析等方法,对处理水产养殖污水潜流湿地中ANAMMOX菌的结构、多样性和丰度进行了研究分析.结果表明,处理水产养殖污水潜流湿地中存在Candidatus brocadia和Candidatus kuenenia这2类已知的ANAMMOX菌和3类未知的ANAMMOX菌,其优势种群为Candidatus brocadia;在不同季节中,秋季潜流湿地中ANAMMOX菌的多样性最高(H',1.21),春季最低(H',0.64);在养殖周期内,潜流湿地中ANAMMOX菌的丰度(以鲜重计,下同)在8.0×104~9.4×106copies·g-1之间,总细菌丰度在7.3×109~9.1×1010copies·g-1之间;在潜流湿地不同层面以及不同季节中,ANAMMOX菌的丰度存在着明显的差异,总细菌丰度则差异不明显,ANAMMOX菌的丰度在不同层面上的差异没有随时间变化的规律性.根据潜流湿地中ANAMMOX菌的分布特征,改变养殖污水布水方式和调整湿地结构可以提高潜流湿地的反硝化效果.该研究为进一步优化潜流湿地结构,提高净化养殖污水效率提供了依据.

Community Characteristics of ANAMMOX Bacteria in Subsurface Flow Constructed Wetland(SSFCW) for Processing of Aquaculture Waster Water

Anaerobic ammonium oxidation (ANAMMOX) is one of the important functions in waste water treatment by subsurface flow constructed wetland(SSFCW), however, there are few studies on ANAMMOX in SSFCW environment at present. The community characteristics of ANAMMOX in the SSFCW of processing aquaculture waste water were explored in this study. In order to analyze the structure, diversity and abundance of ANAMMOX bacteria, several 16S rRNA clone libraries were constructed and real-time PCR targeting specific 16S rRNA genes together with diversity analysis was adopted. The obtained results showed that the SSFCW identified a total of three unknown clusters and two known clusters including Candidatus brocadia and Candidatus kuenenia. The dominant cluster was Candidatus brocadia. The highest diversity levels of ANAMMOX bacteria occurred in autumn(H', 1.21), while the lowest in spring (H', 0.64). The abundance of ANAMMOX bacteria in SSFCW environment ranged from 8.0×10⁴ to 9.4×10⁶ copies ·g^-1 of fresh weight and the copy number of total bacterial 16S rRNA genes ranged from 7.3×10⁹ to 9.1×10¹⁰ copies ·g^-1 of fresh weight during culture cycle. There were significant differences in the ANAMMOX bacteria abundances of different stratum and seasons in SSFCW environment, but the differences in total bacterial abundances were not obvious. In addition, the differences in ANAMMOX bacteria abundances in different stratum and seasons in SSFCW environment were irregular in different culture cycle. According to the distribution characteristics of ANAMMOX bacteria in the wetland, the denitrification effect of SSFCW could be improved by changing the supplying manners of aquaculture wastewater and adjusting the structure of wetland. The research results will provide reference for further optimizing the SSFCW and improving the efficiency of purification.