Use of stable-isotope probing, full-cycle rRNA analysis, and fluorescence in situ hybridization-microautoradiography to study a methanol-fed denitrifying microbial community

Ginige, MP, Hugenholtz, P, Daims, H, Wagner, M, Keller, J and Blackall, LL (2004) Use of stable-isotope probing, full-cycle rRNA analysis, and fluorescence in situ hybridization-microautoradiography to study a methanol-fed denitrifying microbial community. Applied And Environmental Microbiology, 70 1: 588-596. doi:10.1128/AEM.70.1.588-596.2004

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ72779_OA.pdf Full text (open access) application/pdf 1.34MB 0

Author Ginige, MP
Hugenholtz, P
Daims, H
Wagner, M
Keller, J
Blackall, LL
Title Use of stable-isotope probing, full-cycle rRNA analysis, and fluorescence in situ hybridization-microautoradiography to study a methanol-fed denitrifying microbial community
Journal name Applied And Environmental Microbiology   Check publisher's open access policy
ISSN 0099-2240
Publication date 2004
Sub-type Article (original research)
DOI 10.1128/AEM.70.1.588-596.2004
Open Access Status File (Publisher version)
Volume 70
Issue 1
Start page 588
End page 596
Total pages 9
Editor Judy D. Wall
L. Nicholas Ornston
Place of publication United States
Publisher Amercian Society for Microbiology
Collection year 2004
Language eng
Subject C1
270399 Microbiology not elsewhere classified
779999 Other
Abstract A denitrifying microbial consortium was enriched in an anoxically operated, methanol-fed sequencing batch reactor (SBR) fed with a mineral salts medium containing methanol as the sole carbon source and nitrate as the electron acceptor. The SBR was inoculated with sludge from a biological nutrient removal activated sludge plant exhibiting good denitrification. The SBR denitrification rate improved from less than 0.02 mg of NO3-.N mg of mixed-liquor volatile suspended solids (MLVSS)(-1) h(-1) to a steady-state value of 0.06 mg of NO3-.N mg of MLVSS-1 h(-1) over a 7-month operational period. At this time, the enriched microbial community was subjected to stable-isotope probing (SIP) with [C-13] methanol to biomark the DNA of the denitrifiers. The extracted [C-13]DNA and [C-12]DNA from the SIP experiment were separately subjected to full-cycle rRNA analysis. The dominant 16S rRNA gene phylotype (group A clones) in the [C-13]DNA clone library was closely related to those of the obligate methylotrophs Methylobacillus and Methylophilus in the order Methylophilales of the Betaproteobacteria (96 to 97% sequence identities), while the most abundant clone groups in the [C-12]DNA clone library mostly belonged to the family Saprospiraceae in the Bacteroidetes phylum. Oligonucleotide probes for use in fluorescence in situ hybridization (FISH) were designed to specifically target the group A clones and Methylophilales (probes DEN67 and MET1216, respectively) and the Saprospiraceae clones (probe SAP553). Application of these probes to the SBR biomass over the enrichment period demonstrated a strong correlation between the level of SBR denitrification and relative abundance of DEN67-targeted bacteria in the SBR community. By contrast, there was no correlation between the denitrification rate and the relative abundances of the well-known denitrifying genera Hyphomicrobium and Paracoccus or the Saprospiraceae clones visualized by FISH in the SBR biomass. FISH combined with microautoradiography independently confirmed that the DEN67-targeted cells were the dominant bacterial group capable of anoxic [C-14] methanol uptake in the enriched biomass. The well-known denitrification lag period in the methanol-fed SBR was shown to coincide with a lag phase in growth of the DEN67-targeted denitrifying population. We conclude that Methylophilales bacteria are the dominant denitrifiers in our SBR system and likely are important denitrifiers in full-scale methanol-fed denitrifying sludges.
Keyword Biotechnology & Applied Microbiology
Microbiology
Waste-water Treatment
Polyphosphate-accumulating Organisms
Targeted Oligonucleotide Probes
Activated-sludge
Nitrogen Removal
External Carbon
Identification
Denitrification
Bacteria
Ecology
Q-Index Code C1

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 143 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 145 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 15 Aug 2007, 04:19:54 EST