Investigation of an acetate-fed denitrifying microbial community by stable isotope probing, full-cycle rRNA analysis, and fluorescent in situ hybridization-microautoradiography

Ginige, MP, Keller, J and Blackall, LL (2005) Investigation of an acetate-fed denitrifying microbial community by stable isotope probing, full-cycle rRNA analysis, and fluorescent in situ hybridization-microautoradiography. Applied And Environmental Microbiology, 71 12: 8683-8691. doi:10.1128/AEM.71.12.8683-8691.2005

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Author Ginige, MP
Keller, J
Blackall, LL
Title Investigation of an acetate-fed denitrifying microbial community by stable isotope probing, full-cycle rRNA analysis, and fluorescent in situ hybridization-microautoradiography
Journal name Applied And Environmental Microbiology   Check publisher's open access policy
ISSN 0099-2240
Publication date 2005
Sub-type Article (original research)
DOI 10.1128/AEM.71.12.8683-8691.2005
Open Access Status File (Publisher version)
Volume 71
Issue 12
Start page 8683
End page 8691
Total pages 9
Editor Alexander Steinbuechel
Place of publication Washington
Publisher American Society Microbiology
Collection year 2005
Language eng
Subject C1
270399 Microbiology not elsewhere classified
780199 Other
Abstract The acetate-utilizing microbial consortium in a full-scale activated sludge process was investigated without prior enrichment using stable isotope probing (SIP). [C-13]acetate was used in SIP to label the DNA of the denitrifiers. The [C-13]DNA fraction that was extracted was subjected to a full-cycle rRNA analysis. The dominant 16S rRNA gene phylotypes in the C-13 library were closely related to the bacterial families Comamonadaceae and Rhodocyclaceae in the class Betaproteobacteria. Seven oligonucleotide probes for use in fluorescent in situ hybridization (FISH) were designed to specifically target these clones. Application of these probes to the sludge of a continuously fed denitrifying sequencing batch reactor (CFDSBR) operated for 16 days revealed that there was a significant positive correlation between the CFDSBR denitrification rate and the relative abundance of all probe-targeted bacteria in the CFDSBR community. FISH-microautoradiography demonstrated that the DEN581 and DEN124 probe-targeted cells that dominated the CFDSBR were capable of taking Up [C-14] acetate under anoxic conditions. Initially, DEN444 and DEN1454 probe-targeted bacteria also dominated the CFDSBR biomass, but eventually DEN581 and DEN124 probe-targeted bacteria were the dominant bacterial groups. All probe-targeted bacteria assessed in this study were denitrifiers capable of utilizing acetate as a source of carbon. The rapid increase in the number of organisms positively correlated with the immediate increase in denitrification rates observed by plant operators when acetate is used as an external source of carbon to enhance denitrification. We suggest that the impact of bacteria on activated sludge subjected to intermittent acetate supplementation should be assessed prior to the widespread use of acetate in the waste-water industry to enhance denitrification.
Keyword Biotechnology & Applied Microbiology
Microbiology
Waste-water Treatment
Polyphosphate-accumulating Organisms
Biological Phosphorus Removal
Activated-sludge
External Carbon
Nitrogen Removal
Sp Nov.
Bacteria
Denitrification
Identification
Q-Index Code C1

 
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Created: Wed, 15 Aug 2007, 07:00:00 EST