PMID- 17215016
OWN - NLM
STAT- MEDLINE
DCOM- 20070420
LR  - 20191210
IS  - 0043-1354 (Print)
IS  - 0043-1354 (Linking)
VI  - 41
IP  - 4
DP  - 2007 Feb
TI  - Quantification of anaerobic ammonium-oxidizing bacteria in enrichment cultures by 
      real-time PCR.
PG  - 785-94
AB  - The anaerobic ammonium-oxidizing (ANAMMOX) bacteria were enriched from a rotating 
      disk reactor (RDR) biofilm in semi-batch cultures. Based on fluorescence in situ 
      hybridization (FISH) analysis, this enrichment led to a relative population size 
      of 36% ANAMMOX bacteria. Phylogenetic analysis revealed that all the detected 
      clones were related to the previously reported ANAMMOX bacteria, Candidatus 
      Brocadia anammoxidans (AF375994), with 92% sequence similarity. Furthermore, we 
      successfully developed a real-time polymerase chain reaction (PCR) assay to 
      quantify populations of ANAMMOX bacteria in the enrichment cultures. For this 
      real-time PCR assay, PCR primer sets targeting 16S ribosomal RNA genes of ANAMMOX 
      bacteria were designed and used. The quantification range of this assay was 6 
      orders of magnitude, from 8.9x10(1) to 8.9x10(6) copies per PCR, corresponding to 
      the detection limit of 3.6x10(3) target copies mL(-1). A significant correlation 
      was found between the increase in copy numbers of 16S rRNA gene of ANAMMOX 
      bacteria and the increase in nitrogen removal rates in the enrichment cultures. 
      Quantifying ANAMMOX bacterial populations in the enrichment culture made it 
      possible to estimate the doubling time of the enriched ANAMMOX bacteria to be 3.6 
      to 5.4 days. The real-time PCR assay gave comparable population sizes in the 
      enrichment cultures with the FISH results. These results suggest that the 
      real-time PCR assay developed in this study is useful and reliable for 
      quantifying the populations of ANAMMOX bacteria in environmental and engineering 
      samples.
FAU - Tsushima, Ikuo
AU  - Tsushima I
AD  - Department of Urban and Environmental Engineering, Graduate School of 
      Engineering, Hokkaido University, North-13, West-8, Sapporo 060-8628, Japan.
FAU - Kindaichi, Tomonori
AU  - Kindaichi T
FAU - Okabe, Satoshi
AU  - Okabe S
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070109
PL  - England
TA  - Water Res
JT  - Water research
JID - 0105072
RN  - 0 (Nitrites)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - 7664-41-7 (Ammonia)
SB  - IM
MH  - Ammonia/*metabolism
MH  - Bacteria, Anaerobic/genetics/*isolation & purification/*metabolism
MH  - Biofilms
MH  - Biomass
MH  - Bioreactors
MH  - In Situ Hybridization, Fluorescence
MH  - Nitrites/metabolism
MH  - Oxidation-Reduction
MH  - Phylogeny
MH  - *Polymerase Chain Reaction
MH  - RNA, Ribosomal, 16S/metabolism
MH  - Time Factors
EDAT- 2007/01/12 09:00
MHDA- 2007/04/21 09:00
CRDT- 2007/01/12 09:00
PHST- 2006/07/10 00:00 [received]
PHST- 2006/10/21 00:00 [revised]
PHST- 2006/11/08 00:00 [accepted]
PHST- 2007/01/12 09:00 [pubmed]
PHST- 2007/04/21 09:00 [medline]
PHST- 2007/01/12 09:00 [entrez]
AID - S0043-1354(06)00631-2 [pii]
AID - 10.1016/j.watres.2006.11.024 [doi]
PST - ppublish
SO  - Water Res. 2007 Feb;41(4):785-94. doi: 10.1016/j.watres.2006.11.024. Epub 2007 
      Jan 9.