Document Detail


Real-time PCR assay for the simultaneous quantification of nitrifying and denitrifying bacteria in activated sludge.
MedLine Citation:
PMID:  17256118     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In order to improve wastewater treatment processes, a need exists for tools that rapidly give detailed insight into the community structure of activated sludge, supplementary to chemical and physical data. In this study, the advantages of microarrays and quantitative polymerase chin reaction (PCR) methods were combined into a real-time PCR assay that allows the simultaneous quantification of phylogenetic and functional genes involved in nitrification and denitrification processes. Simultaneous quantification was possible along a 5-log dynamic range and with high linear correlation (R (2) > 0.98). The specificity of the assay was confirmed by cloning and sequencing analyses of PCR amplicons obtained from activated sludge. The real-time assay was validated on mixed liquid samples of different treatment plants, which varied in nitrogen removal rate. The abundance of ammonia oxidizers was in the order of magnitude of 10(6) down to 10(4) ml(-1), whereas nitrite oxidizers were less abundant (10(3)-10(1) order of magnitude). The results were in correspondence with the nitrite oxidation rate in the sludge types. As for the nirS, nirK, and nosZ gene copy numbers, their abundance was generally in the order of magnitude of 10(8)-10(5). When sludge samples were subjected to lab-scale perturbations, a decrease in nitrification rate was reflected within 18 h in the copy numbers of nitrifier genes (decrease with 1 to 5 log units), whereas denitrification genes remained rather unaffected. These results demonstrate that the method is a fast and accurate tool for the analysis of the (de)nitrifying community structure and size in both natural and engineered environmental samples.
Authors:
Joke Geets; Michaël de Cooman; Lieven Wittebolle; Kim Heylen; Bram Vanparys; Paul De Vos; Willy Verstraete; Nico Boon
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-01-26
Journal Detail:
Title:  Applied microbiology and biotechnology     Volume:  75     ISSN:  0175-7598     ISO Abbreviation:  Appl. Microbiol. Biotechnol.     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-04-26     Completed Date:  2007-07-19     Revised Date:  2012-05-28    
Medline Journal Info:
Nlm Unique ID:  8406612     Medline TA:  Appl Microbiol Biotechnol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  211-21     Citation Subset:  IM    
Affiliation:
Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Coupure Links 653, Ghent, Belgium.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/DQ857294;  DQ857295;  DQ857296;  DQ857297;  DQ857298;  DQ857299;  DQ857300;  DQ857301;  DQ857302;  DQ857303;  DQ857304;  DQ857305;  DQ857306;  DQ857307;  DQ857308;  DQ857309;  DQ857310;  DQ857311;  DQ857312;  DQ857313;  DQ857314;  DQ857315
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MeSH Terms
Descriptor/Qualifier:
Bacteria / genetics,  isolation & purification
DNA, Bacterial / analysis,  isolation & purification
Molecular Sequence Data
Nitrate Reductase / genetics
Nitrite Reductases / genetics
Nitrites / metabolism*
Oligonucleotide Array Sequence Analysis / methods
Organic Chemicals
Oxidoreductases / genetics
Polymerase Chain Reaction / methods*
RNA, Ribosomal, 16S / genetics
Sequence Analysis, DNA
Sewage / microbiology*
Waste Disposal, Fluid / methods*
Chemical
Reg. No./Substance:
0/DNA, Bacterial; 0/Nitrites; 0/Organic Chemicals; 0/RNA, Ribosomal, 16S; 0/Sewage; 163795-75-3/SYBR Green I; EC 1.-/Oxidoreductases; EC 1.7.-/Nitrite Reductases; EC 1.7.2.4/nitrous oxide reductase; EC 1.7.99.4/Nitrate Reductase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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