人工湿地高效好氧反硝化菌的分离鉴定及反硝化特性研究_王莹

农业环境科学学报2010,29(6):1193-1198JournalofAgro-EnvironmentScience摘要：从增氧型复合垂直流人工湿地中采集样品，利用间歇曝气法富集好氧反硝化菌，并进行分离纯化，共得到10株好氧反硝化菌。其中编号为B13的菌株在初始硝态氮含量为277.23mg·L-1、碳氮比为5的条件下，24h的硝态氮去除率达92.80%，亚硝态氮积累只有12.57mg·L-1，脱氮速率达到20.58mg·L-1·h-1。16SrDNA序列分析表明，该菌与Pseudomonasstutzeri同源性达100%。选用四因素三水平L9（34）正交试验表设计实验，通过测定对硝态氮去除能力和亚硝态氮的积累量，研究碳源、碳氮比（C/N）、pH以及溶解氧含量（DO）4种不同因素对B13号菌株好氧反硝化性能的影响。结果表明，该菌株对硝态氮的去除率最大可达99.88%，几乎没有亚硝态氮积累。对硝态氮去除率影响最大的因素为碳氮比，其次为pH，溶解氧含量和碳源。对应的最优条件是碳源为葡萄糖，碳氮比为10，pH为9，溶解氧含量为1.84～3.57mg·L-1。关键词：人工湿地；好氧反硝化细菌；反硝化活性；系统发育分析中图分类号：X172文献标志码：A文章编号：1672-2043(2010)06-1193-06人工湿地高效好氧反硝化菌的分离鉴定及反硝化特性研究王莹1，2，周巧红1，梁威1，徐栋1，蔡林林1，2，陶敏1，2，吴振斌1（1.中国科学院水生生物研究所淡水生态与生物技术国家重点实验室，武汉430072；2.中国科学院研究生院，北京100039）IsolationandIdentificationofaHigh-efficiencyAerobicDenitrifierandItsDenitrifyingCharacteristicinConstructedWetlandWANGYing1,2,ZHOUQiao-hong1,LIANGWei1,XUDong1,CAILin-lin1,2,TAOMin1,2,WUZhen-bin1（1.StateKeyLaboratoryofFreshwaterEcologyandBiotechnology,InstituteofHydrobiology,ChineseAcademyofSciences,Wuhan430072,China;2.GraduateUniversity,ChineseAcademyofSciences,Beijing100049,China）Abstract：TheidentificationofaAerobicdenitrifieranditspracticabilityforaerobicwastewatertreatmentinIntegratedVertical-flowCon－structedWetland（IVCW）werestudiedinthispaper.SampleswerecollectedfromanaeratedIVCW.Afterenrichmentbyintermittentaera－tion,isolationandpurificationinturn,10strainsofaerobicdenitrifierswereobtainedandtheirdenitrifyingcapabilitywerestudiedinbatchcultureunderaerobiccondition.ResultsshowedthatthemostefficientstrainwasB13.Withinitialnitrateconcentrationof277.23mg·L-1,C/N（carbon/nitrogen）ratioof5andanincubationtimeof24h,nitrateremovalratereached92.80%andnitriteaccumulationwasinalowcon－centrationof12.57mg·L-1.Themaximumremovalrateofnitratewas20.58mg·L-1·h-1.Phylogeneticanalysisbasedonpartial16srDNAshowedthatstainB13had100%sequencesimilaritywithPseudomonasstutzeri.AnorthogonalexperimentbyfourfactorswiththreelevelsL9（34）wasdesignedtoexplorethebestaerobicdenitrificationconditionforstrainB13.Thefourfactorswer