高含硫气田采出水生化处理的微生物构效关系

Microbial Community and Activity Relationship in Biological Treatment of Production Water from High Sour Gas Field

为了探究高盐、高重铬酸盐法化学需氧量(COD_(cr))、高氨氮采出水生化处理的关键功能菌群及代谢机理,在生产规模缺氧/好氧池-膜生物反应池(anoxic/oxic pond-membrane bioreactor,A/O-MBR),使用Illumina Miseq高通量测序和实时荧光定量多聚核苷酸链式反应(real-time quantitative polymerase chain reaction,qPCR)方法,检测了细菌、氨氧化细菌(ammonia oxidizing bacteria,AOB)和氨氧化古菌(ammonia oxidizing archaea,AOA)的组成和含量。A/O-MBR出水分析结果显示COD_(cr)平均211 mg/L,去除率72.7%;氨氮分为低/高去除率(LS/HS)两个阶段,去除率分别为5.7%和70.8%。氨氮全部氧化为亚硝酸根;但亚硝酸根没有进一步转化为硝酸根,而是部分通过反硝化转化为氮气。细菌组成揭示关键功能菌群:涉及CODcr去除的主要有林杆菌(Limnobacter)、陶厄氏菌(Thauera)、伯克氏菌(Burkholderia)、水微菌(Aquamicrobium)、藤黄单胞菌(Luteimonas)、砂单胞菌(Arenimonas),涉及反硝化的主要有斯塔普氏菌(Stappia)、生丝微菌(Hyphomicrobium)和噬甲基菌(Methylophaga)。AOB优势菌是亚硝化单胞菌(Nitrosomonas),占比高达96.6%。AOA主要有亚硝化球菌(Nitrososphaera)、亚硝化短小杆菌(Nitrosopumilus)和Nitrosocosmicus,均率属于奇古菌门(Thaumarchaeota)。LS和HS样品细菌含量为2.83×10^(9)~4.01×10^(9)拷贝/g污泥,细菌丰富是高效生化处理的基础;AOB含量分别为7.27×10^(7)~8.47×10^(7)拷贝/g污泥和6.79×10^(8)~17.2×10^(8)拷贝/g污泥,差异显著,是氨氮去除率差异的决定因素;AOA含量为4.06×10^(4)~8.88×10^(4)拷贝/g污泥,均极低,揭示其不是氨氮去除的关键菌群。这些发现可为提高含硫气田高含盐采出水处理效率和稳定性奠定基础。

To reveal the key functional microbes and metabolic mechanisms in biological treatment of hypersaline production water with high levels of chemical oxygen demand determined by dichromate method(COD_(cr))and ammonia.The illumina Miseq and real-time quantitative polymerase chain reaction(qPCR)were employed to detect the community and quantity of bacteria,ammonia oxidizing bacteria(AOB)and ammonia oxidizing archaea(AOA)within a full-scale A/O-MBR(anoxic/oxic pond-membrane bioreactor).Results showed that the average COD_(cr) of the effluent is 211 mg/L,and the removal rate is 72.7%.Ammonia demonstrated a two-stage pattern of low/high removal rate(LS/HS),corresponding to 5.7%and 70.8%,respectively.Ammonia is completely transformed into nitrite.Nitrite is not further oxidized into nitrate,but partly reduced to nitrogen gas by denitrifying bacteria.Bacterial community reveals that the key bacteria involved in COD_(cr) removal are Limnobacter,Thauera,Burkholderia,Aquamicrobium,Luteimonas and Arenimonas,and the key denitrifying bacteria are Stappia,Hyphomicrobium and Methylophaga.The dominant one in AOB is Nitrosomonas,accounting for more than 96.6%.AOA is primarily composed with Nitrososphaera,Nitrosopumilus and Nitrosocosmicus,all affiliated to Thaumarchaeota.The bacteria content is 2.83×10^(9)~4.01×10^(9) copies/g sludge for both LS and HS samples,which result in the high efficiency of biological treatment.The AOB content is 7.27×10^(7)~8.47×10^(7) copies/g sludge,6.79×10^(8)~17.2×10^(8) copies/g sludge,respectively for LS and HS samples,and this an order of magnitude difference led to LS/HS two-stage pattern.The AOA copies/g sludge 4.06×10^(4)~8.88×10^(4) copies/g sludge,and this less quantity revealed that those contributions to ammonia removal could be neglected.These results could be used for improving the efficiency and stability for biological treatment of hypersaline water from sour gas fields.