城市污水处理系统的菌群分析和除氮基因工程出发菌的选择马展

:1005-376X(2004)02-0083-02【　　】马展,张德纯(重庆医科大学检验系临床微生物教研室,重庆　400016)　　　:,。:,,—,。:,、、,23%、16%、16%12%。,、。:,,。:;:Q78;X703　　　　:AMicrofloraanalyseinactivesludgeofsewagetreatmentandcloneandexpressnapgeneclusterinPseudomonasMAZhan,ZHANGDe-chun(DepartmentofClinicalMicrobiologyChongqingUniversityofMedicalScience,Chongqing410016,China)Abstract:Objective:Toexplicatethecompositionofmicroflorainactivesludgeofsewagetreatment.Toob-tainproperstrainsfittingforgenesmanipulatingtoenhancetheabilityofdenitrification.Methods:Firstofall,wecountedthepopulationofbacteriaintheactivesludge.Usingtheirbiochemicalandgrowthpropertiesidentifythosestrainswehadisolated.Results:Wenoticedthatdominativebacteriaintheactivesludgeofdenitrifyingre-actorwerePseudomonas(23%),Acinetobacter(12%),Moraxella(16%)andEnterobacter(16%).Chosingtwoofthembasedontheirfeaturesaboutdenitrationandresistancetoantibiotics.Conclusion:ThosetwobacteriawereidentifiedasPseudomonasstutzeriandPseudomonasmendocina.Keywords:Denitration;Pseudomonas　　,、、,[1]。,,,。,,,,。1　1.1　培养基和试剂1.1.1　模拟混合污水琼脂培养基　70mg,170mg,110mg,NH4Cl180mg,3.2g,Na2CO3110mg,80mg,K2HPO430mg,1000ml。,pH7.4,15g,115℃25min。:5dBOD5:190～280mg/L,:90～100mg/L,5.0～8.0mg/L。1.1.2　、[2],KIA、MHA、。1.1.3　1.2　主要仪器　(UV-265),(Rosi1000),(Millipore),(KubotaKR/702)。:2003-11-12:(1974-),,,,1.3　方法　1.3.1　标本的获取、预处理和接种　A2/0-SBR25ml。15g,20min。0.9%,1050.1ml90mm,28℃48h,1,。1.3.2　细菌鉴定　《》[3]《》[4]。1.3.3　假单胞各菌株脱氮能力测定　3mlTSB28℃300r/min18h。,3,A600≈0.6。10%,2/3。28℃180r/min8h,10000r/min10,GB11894-89[5]。1　(μg/ml)12341530601207.515306025501002001530601201225501001.3.4　候选菌的药物敏感分析　、、、。(1)1MHA。(2)TSB,300r/min36h,83　20044162　ChineseJournalofMicroecology,April2004,Vol.16No.2DOI:10.13381/j.cnki.cjm.2004.02.009MHA,28℃48h。(3),。2　2.1　污泥标本的菌群构成　203(96+107)。1,22,22.9%;15,15.6%;15,15.6%;12,12.5%;9,9.4%;2,2.1%;3,3.1%;9,9.4%;3,3.1%;2,2.1%;4;4.2%。22,14,6,1,1。2.2　候选菌的脱氮活性　8hP1、P7、P11、P12,9.2mg/L、16.7mg/L、4.4mg/L、57.1mg/L。2.3　候选菌的详细生长鉴定结果2　O/F　DNAP10+-+-+++-+1+P70+-+--++--1-+P110++-+--+--1+--P120++-+-++-+1-+2.4　候选菌在5种抗生素平板上生长的最低浓度3　(μg/ml)　P112060　20060100P71560　503025P113030　2001525P1212060　200301003　,。。,,,、、[1]。[6],,、、,、,;[7-9]。、、、[1],,、、[10]。,、、,,,、、、、、,[6,11]。,,、。,,。,[11],,;,[12,13]。,。,,。P7P11,,,[14],。,,。。:[1].[M].:,1987.[2].[M].:,1999.[3],.[M].:,2001.[4]R.E.,N.E..[M].8::,1984.[5].:[P].GB11894-89.[6],.[M].:,1998.[7]ZumftWG.Cellbiologyandmolecularbasisofdenitrification[J].Mi-crobiolMolBiolRev,1997,61(4):533-616.[8]EllingtonMJ,BhakooKK,SawersG.Hierarchyofcarbonsourcese-lectioninParacoccuspantotrophus:strictcorrelationbetweenreductionstateofthecarbonsubstrateandaerobicexpressionofthenapoperon[J].JBacteriol,2002,184(17):4767-4774.[9]RonchelMC,RamosC,JensenLB.Constructionandbehaviorofbio-logicallycontainedbacteriaforenvironmentalapplicationsinbioreme-diation[J].ApplEnvironMicrobiol,1995,61(8):2990-2994.[10].[M].:,2002.[11]KubaT.OccurrencedenitrifyingphosphorusremovingbacteriainmodifiedUCT-typewastewatertreatmentplants[J].WatRes,1997,31(4):777-786.[12],.[J]..2003,15(3):186-187.[13]SakaguchiK.VectorsforgenecloninginPseudomonasandtheirap-plications[J].CurrTopMicrobiolImmunol,1982,96:31-45.[14]MarxJL.Assessingtherisksofmicrobialrelease[J].Science,1987,237(4821):1413-1417.84　20044162　ChineseJournalofMicroecology,April2004,Vol.16No.2