海洋硝化菌群的富集驯化和筛选的初步研究

291Vol.29,No.120082MARINEFISHERIESRESEARCHFeb.,2008()(IMJQ10000043)*Email:[e-mail],Tel:[phone]:20061101;:20070430:(1972),,(),,Email:[e-mail],Tel:[phone]王光玉1,2马放2*魏琦峰1徐仲1(1(),264209)(2,150090)以循环养殖废水为介质,利用不断提高氨氮浓度的方法富集驯化海洋硝化菌群经过70d的富集驯化,海洋异养细菌数目变化不大,氨氧化细菌和亚硝酸氧化细菌分别增加了43倍和29倍驯化液中氨氧化能力提高55倍,造成了亚硝酸盐和硝酸盐的积累利用选择性培养基从驯化液中分离到7株硝化菌,通过菌落形态和菌体形态观察对分离的菌株进行了初步的鉴定通过硝化能力测试,筛选出硝化能力强的3株细菌水产养殖富集氨氮硝化菌鉴定X172A10007075(2008)01007605Studyonenrichment,domesticationandscreeningofmarinenitrobacteriaWANGGuangyu1,2MAFang2*WEIQifeng1XUZhong1(1TheOceanCollegeofHarbinInstituteofTechnology,Weihai264209)(2SchoolofMunicipalandEnvironmentalEngineeringofHIT,150090)ABSTRACTThenitrobacteriainwastewaterfromrecirculationaquaculturesystemswereenrichedbycontinuouslyincreasingtheconcentrationofammonia.After70denrichment,nosignificantchangeinthetotalnumberofmarineheterotrophicbacteriawasfound,whilethenumberofnitrosomonasandnitrobacteriaincreased43and29times,respectively.Theabilityofnitrationwasenhanced55times,resultingintheaccumulationofnitriteandnitrate.Sevenspeciesofnitrosomonasornitrobacteriawereobtainedbyselectivemedium,andprimaryidentificationwasdonebythesurfacecharacterofthecoloniesandthebacteria.Threespeciesofthebacteriawereselectedfortheirhighnitrificationability.KEYWORDSAquacultureEnrichmentAmmoniaNitrobacteriaIdentification,,,,;,1:,,,,,,(1999;Sungkookimetal.2000;JaekoanSeoetal.2001),,,,,,,,(2002;2000)11.1160cm40cm30cm,30L,252,,DO6ml,1mol/LNaOHpH80~86,,10mg/L,,70d(2002;2004)1.2,pHCODDO9,,1.3(1999)GB173784,COD,,,,,(HANNAHI98128)pH,,(HANNAHI9142)DOGB173787,2216E,()(2002),MPN2514d,,2520d,,;(1000),,10%1.4,,,2510d,,77291.5NH+4NNO2N50mg/L,18ml10ml,3,25,100r/min5d,,,22.1,COD,123773,,pH,,,,pH809~845,84,pH81,(2004)(1),101128mg/L62031mg/L,55,,,,,,1679mg/L,0321mg/L(2),,,(SunkooKimetal.2000;Jaekoanseoetal.2001)(2003;2005)2.2781:3Fig.3Microorganismprofileduringtheexperiment,,(1996),3,,4329,,2.3,,14,31Table1ColonialfeaturesofnitrifyingmediumNumberSpecies(cm)Size(cm)ColorTransparencyProtuberanceVergeQuantity1-1#3cm481-2#2~3221-3#1~2361-4#0.5302-1#2~3362-2#1.5~2.5362-3#0.56024,,,2(1988;Vanginkeletal.1983)2Table2MorphologicalfeaturesofnitrobacteriaNumberG+orG-(m)SizeShapeArraymodePrimaryidentification11#G-0.4~0.612#G-(0.40.6)~(0.91.2)13#G-0.5~(1.01.2)14#G-0.8~1.0,21#G-1.522#G-(0.40.5)~(1.01.2)23#G-0.5~0.82.51ml1ml50ml,7929NO-2N,NO-2N,1,1ml2,45,14#,11#12#,3,21#,(1999)4Fig.4Abilityofammoniumoxidizingbacteria5Fig.5Nitriteoxidizingabilityofnitrobacteria,,.2002..:,,.2002..,23(12):46~48,.1999..,29(1):21~24,,.2004..,23(3):540~544,,.2003..,4(11):44~47,,.1996.,15(3):18~20,,.2000..,19(5):225~227,,.2002..,14(6):348~350,,,.2005..,14(1):26~29.1998.,,,.1999..,27(3):351~354J.G..1988..:JaeKoanSeo,IlHyongJung,MiRyungKim,ByongJinKim,SooWanNam,andSungKooKim.2001.NitrificationperformanceofnitrifiersimmobilizedinPVAforamarinerecirculatingaquariumsystem.AquaculturalEngineering,4:181~194SungKooKim,InsooKong,ByungHunLee,LimseokKang,MinGyuLee,andKuenHackSuh.2000.RemovalofammoniumNfromarecirculationaquaculturalsystemusinganimmobilizednitrifier.AquaculturalEngineering,21:139~150Vanginkel,C.G.,Tramper,J.,andLuyben,K.C.A.M.1983.CharacterizationofNitrosomonaseuropaeaimmobilizedincalciumalginate.EnzymeandMicrobialTechnology,(5):297~30380