ARS | Publication request: Detection of Sulfate Reducing Bacteria in Stored Swine Manure Using Quantitative, Real-Time Pcr Analysis

Submitted to: Microbial Ecology International Symposium
Publication Type: Abstract
Publication Acceptance Date: August 27, 2004
Publication Date: August 27, 2004
Citation: Cook, K.L., Whitehead, T.R., Cotta, M.A. 2004. Detection of sulfate reducing bacteria in stored swine manure using quantitative, real-time pcr analysis [abstract]. Microbial Ecology International Symposium. Paper No. 226.

Technical Abstract: Sulfate-reducing bacteria (SRB) are a diverse group of anaerobic microorganisms responsible for production of hydrogen sulfide. Hydrogen sulfide is one of the more odorous compounds produced as a consequence of incomplete digestion of stored swine manure. However, the concentrations of specific groups of SRB in underground pits, used for manure storage, have not been determined. To address this question, quantitative real-time PCR was used to target the dissimilatory sulfite reductase A (dsrA) gene of SRB. A TaqMan probe targeted the dsrA gene of all the cloned SRB. Primers specifically amplified the dsrA gene sequences of Desulfobulbus-like (Group 1) or Desulfovibrio-like (Group 2) SRB matching slurry clones or Desulfovibrio-like (Group 3) SRB from slurry enrichment cultures. Manure samples were taken at different times of the year, and the concentration of these SRB was evaluated. Results show that the concentration of Group 1 (1.0 X 10**4 to 2.0 X 10**8 dsrA copies/mL slurry), 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 varied over time and between swine houses on the same farm. 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 (2.0 X 10**7 to 6.8 X 10**9 16S rDNA copies/mL slurry). These results suggest that the concentration of SRB is highly variable and may be dependent on both physical and seasonal parameters.