|Cook, Kimberly - Kim|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 6/7/2004
Publication Date: 6/7/2004
Citation: Cook, K.L., Cotta, M.A., Whitehead, T.R. 2004. Variability in the concentration of sulfate-reducing bacteria in swine manure and feces as determined using a real-time PCR assay [abstract]. Reproduction Nutrition Development. 44(1):S6.
Technical Abstract: Real-time quantitative PCR was used to target sulfate-reducing bacteria (SRB) in order to evaluate their concentration in stored swine manure and swine feces. Real-time quantitative PCR permits rapid, sensitive detection of target organisms using fluorescently labeled probes. As the PCR product is produced, fluorescence signal from the probe increases in proportion to the starting concentration of the target sequence. In this case, SRB were targeted using a common probe and three different sets of primers designed to specifically amplify the dsrA gene sequences of: Desulfobulbus-like (Group 1) or Desulfovibrio-like (Group 2) SRB matching slurry clones or Desulfovibrio-like (Group 3) SRB matching slurry enrichment culture clones and isolates. Results indicate that Group 1 SRB were more common in pit slurry (concentrations ranging from 1.0 X 10**4 to 2.0 X 10**8 dsrA copies/mL slurry). Concentrations of Group 2 (4.5 X 10**3 to 1.0 X 10**6 dsrA copies/mL slurry) and Group 3 (<1 X 10**3 to 4.3 X 10**6 dsrA copies/mL slurry) SRB were lower. Group 3 SRB were greater than 65% of the total population in the enrichment cultures. However, the SRB population was less than one percent of the total slurry population as determined by 16S rDNA analysis. Results suggest that this real-time PCR assay is sensitive and specific for detection of SRB in stored swine manure.