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(Anammox, Anaerobic Ammonia Oxidation)(NH4+)(NO2-)(N2)0 ml/min10 ml/min100 ml/minPropidiummonoazide (PMA)-(PCR-DGGE)
(CO2)(NH4+)(NO2-)(N2)(Jettenet al., 2001) (Anaerobic AmmoniaOxidation, Anammox)NH4+ + NO2- N2 + 2H2O
Ethidium monoazide (EMA)Propidium monoazide(PMA)EMA PMAEMA PMA (Pan et al.,2007)PMA (Nockeret al., 2006Flekna et al., 2007Cawthorn and Witthuhn, 2008)Propidium monoazide (PMA)PCR-DGGENocker et al., 2006
1. 10000rpm 1 (pellet)1PBS 10000rpm 1 pellet2. Propidium Monoazide (PMA) PMA 20 % dimethyl sulfoxide20 mM2.5 L PMA500 L 5 min650W 2~4 min10000rpm 2 (Nocker et al., 2009)
3. DNA UltraCleanTM Soil DNA Isolation Kit (Mo Bio, USA)DNA DNA PCR PLA46f Amx368r amoA gene (1F vs2R)(Neef et al., 1998Schmid et al., 2003)(Rotthauwe et al., 1997)PCR 1.5%(Agarose gel)Safe ViewerTM Nucleic Acid Stain 30 4. DNA DGGE 6 % (wt/vol) acrylamide/bis30% ~ 55 %PCR Anammox (Dcode gene System, Bio-Rad)80 60 12 Safe ViewerTM Nucleic Acid Stain PCR-DGGE -DNA PCR-DGGE
5. NH4+-NNO2--NNO3--N Ion Chromatography, IC DioNEX-DX100AS-4ANaCO3 NaHCO3CS-15H2SO4
1. Propidium monoazide (PMA)(100 30 min 70 %30 min)PMA PLA46 AMX368 (1)agarose gel377 bp(1 )PMA PMA DNA PCR DGGE (2)PMA
2. UASB (56 )(115 )DGGE (1) 0ml/min 0ml/min 70 mg/L12 50 mg/L30 %180 mg/L150 mg/L20 %0.1 mg/L44 ml(3)
0ml/min 4 Candidatus Brocadia anammoxidansCandidatus Brocadiasp.Candidatus Kuenenia stuttgartiensisCOD 97 mg/L
(2) 10ml/min 10 ml/min 60mg/L0 mg/L100 %120 mg/L3 mg/L97 %30 mg/L38 ml(5)12 10ml/min 6 Candidatus Brocadia anammoxidansCandidatus Brocadiasp.Candidatus Kuenenia stuttgartiensis(10 ml/min)
(3) 100ml/min 100 ml/min 50 mg/L12 13 mg/L74 %130 mg/L45 mg/L 65 %30mg/L1 ml(7)ORP 200 mV2 mg/L120 pH 7 150 ORP 50 mV 169 agarose gel (8)
100ml/min 9 Candidatus Brocadia anammoxidans CandidatusBrocadia sp.Candidatus Kuenenia stuttgartiensis(Arrojo et al., 2008)ORP
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1. 0 ml/min COD 2. 10 ml/min 3. 100 ml/min ORP 4. 10 ml/min