短程硝化ANAMMOX反应器处省略市污水过程中脱氮微生物的群落解析王惠

29120121JournalofGraduateUniversityofChineseAcademyofSciencesVol．29JanuaryNo．12012*2007BAD87B12KSCX2-YW-G-054[e-mail]1002-1175201201-0038-09-ANAMMOX*1223421110004921000223100029447500020114122011420WangHYiPZhangSJetal．Thefunctionalmicrobialcommunitycompositionanalysisinthelaboratory-scalestablepartialnitrifying-ANAMMOXmunicipalwastewaterreactorJ．JournalofGraduateUniversityofChineseAcademyofSciences201229138-46．-．80%．．Nitrosomonaseuropaea/NitrosococcusmobilisNitrobactervulgarisNitrospiradefluviiKueneniastuttgartiensis．Proteobacteria、ChloroflexiBacteroidetes/Chlorobianammox．Q89ANAMMOXNO－2．、、、1-2．、1∶1.3．-2CANONSHARON-ANAMMOX3．、4-5．．DO-、、．．1-ANAMMOX11.11UASB1．28L74cmDO≤0.2mg/L．20～29℃144L/dHRT4.6h75%SRT200d．170cm8cm8.5L．40cm、．30±0.2℃142L/dHRT1.4h．A/O/21．11pHCOD/mg/LTN/mg/LNH+4—N/mg/LNO－2—N/mg/LNO－3—N/mg/LTP/mg/L7.5～8.07.723.4～92.851.645.6～60.751.443.0～54.749.30～0.10.050.1～1.40.70.12～0.820.35．2120d．80%．－20℃．1.21CODN-1--pHpHWTWDOWTWMLSS．NO－2—N/NO－2—N+NO－3—N．2DNA-PCRFastDNADNATsushimaKindaichietal．2007．PCR2．PCR50μL5μLPCR4μL0.25mmol/LdNTPs1μL10μmol/L1μL10μmol/L0.25μL1.25UTaqDNA2μL9329DNA．95℃5min95℃1min1min72℃1.5min3572℃10min．1%PCR．2PCR5'-3'/℃16SrRNA27fAGAGTTTGATCCTGGCTCAG55～6061492rGGWTACCTTGTTACGACTT6AOBamoAamoA-1FGGGGTTTCTACTGGTGGT577-8amoA-2RCCCCTCKGSAAAGCCTTCTTC7-8Nitrobactersp．16SrRNAFGPS872CTAAAACTCAAAGGAATTGA517-9FGPS1269TTTTTTGAGATTTGCTAG7-9Nitrospirasp．16SrRNA341fCCTACGGGAGGCAGCAG537-9NTSPArCGTTATCCTGGGCAGTCCTT7-916SrRNAAmx368TTCGCAATGCCCGAAAGG5510-11Amx820AAAACCCCTCTACTTAGTGCCC10-113FermentasMBIFermentasPCR．pGEM-TEasyPromegaDH5Takara．50mg/L、X-GelIPTG37℃15h．M13F5'-GTAAAACGACGGCCAG-3'M13R5'-CAGGAAACAGCTATGAC-3'．M13PCRABI3730．Bellerophonchimeraehttp∥comp-bio．anu．edu．au/bellerophon/bellerophon．pl．MOTHUR97%OTU．OTUFunGeneFunctionalgenepipeline＆repositoryGenBankBLASThttp∥fungene．cme．msu．edu/index．sprhttp∥．ncbi．nlm．nih．gov/．MEGA4neighbor-joiningalgorithmJukes-CantorSchlossandHandelsman2005．22.12、、-120d．．50mg/L10mg/L0.05mg/L10mg/L91.2%96.3%．NH+4—N/NO－2—N1.0．．0.5mg/L0.9～11.0mg/L≤10mg/L2mg/L．80%．、、2．041-ANAMMOXDO≤0.2mg/LNOBAOB．Hanaki12DO0.5mg/L．/DO．15%DO≤0.2mg/LNOBAOB13．2.2AmoAAOB3．62≥97%9OTU．amoANitrosomonasNitrosococcusNitrosospira．97%OTU1～OTU5OTU7～OTU8Nitrosomonaseuropaea95%～99%．3%OTU9OTU6Nitrosococcusmobilis99%．≤0.2mg/LNitrosomonaseuropaea/Nitrosococcusmobiliscluster．3AOBNitrosomonaseuropaea．HelmerK14N．europaea．ParkHD15N．europaeaO20.12～0.24mg/L．GiesekeA16DO≤O21mg/LN．europaea/eutrophaN．mobilisN．europaea/eutropha．N．mobilisN．europaea/eutrophaN．mobilisN．europaea/eutropha．NitrobacterNitrospira．NitrobacterNitrospira16SNOB4．49Nitrobacter≥97%OTU91429OTU44Nitrospira≥97%OTU1OTU．50%Nitrobacter16SNitrobactervulgaris≥96%50%Nitrobacter16SNitrobacterBradyrhizobiumNitrobacterNitrobacterNitrobactervulgaris．Nitrospira16SrRNANitrospiradefluvii≥98%．NitrobactervulgarisNitrospiradefluvii．4NOBNitrobacterNitrospiraNitrospiraNitrobacter．Nitrospira、、、817-20．Nitrobacter21-22．NitrobacterNitrobactervulgariNOB．Nitrobactervulgari23．Nitrobactervulgari．NitrospiradefluviiNitrospirasublineageI、182024．NitrospiradefluviiNXR、20．Nitrospiradefluvii．AOBNOB．AhnJH8．AOBNOB．2.3anammox16SrRNA．34anammox≥97%4OTU5．65%anammox16SrRNAKueneniastuttgartiensis．OTU2OTU315%CandidateJetteniaasiaticaCandidateBrocadiafulgida98%96%．KueneniastuttgartiensisANAMMOX．25-27．241-ANAMMOXBrocadiafulgidaAnammoxoglobuspropionicus．Scalinduaspp．27-30．Kueneniastuttgartiensis．2000SchmidM31CandidatusKueneniastuttgartiensis．CandidatusKueneniastuttgartiensisCandidatusKueneniastuttgartiensis32．5ANAMMOXanammox143OTU6．86OTU41.2%14.7%Chloroflexi16.8%Bacteroidetes/Chlorobi7.7%Planctomycetales．25.9%β-8.4%γ-4.9%σ-2%α-．B110β-Acidovoraxsp．≥97%33．、Chloroflexi、Bacteroidetes/ChlorobianammoxAcidovoraxsp．β-、、34-35anammox3637Bacteroidetes．anammox．．31≤0.2mg/L120d-80%．2PCR-AOB、NOB、anammoxNitrosomonaseuropaea/NitrosococcusmobilisAOBNitrobactervulgarisNitrospiradefluviiNOBKueneniastuttgartiensis．．3PCR-、ChloroflexiBacteroidetes/Chlorobianammox3429anammox．6ANAMMOX1JettenMSMStrousMVandePas-SchooneKTetal．TheanaerobicoxidationofammoniumJ．FEMSMicrobiologyReviews1998225421-437．2VanDongenUJettenMSVanLoosdrechtMC．TheSHARON-anammoxprocessfortreatmentofammoniumrichwastewaterJ．WaterSciTechnol2001441153-160．3SunSPNacherCPIMerkeyBetal．EffectivebiologicalnitrogenremovaltreatmentprocessesfordomesticwastewaterswithlowC/NratiosAreviewJ．EnvironmentalEngineeringScience2010272111-126．4LamsamALaohaprapanonSAnnachhatreAP．CombinedactivatedsludgewithpartialnitrificationAS/PNandanammoxprocessesfortreatmentofseafoodprocessingwastewaterJ．JEnvironSciHealthAToxHazardSubstEnvironEng200843101198-1208．5YamamotoTTakakiKKoyamaTetal．Novelpartialnitritationtreatmentforanaerobicdigestionliquorofswinewastewaterusingswim-bedtechnologyJ．JournalofBioscienceandBioengineering20061026497-503．6BrofftJEMcArthurJVShimketsLJ．RecoveryofnovelbacterialdiversityfromaforestedwetlandimpactedbyrejectcoalJ．EnvironmentalMicrobiology2002411764-769．441-ANAMMOX7GeetsJDeCoomanMWittebolleLetal．Real-timePCRa