Submitted to: Microbial Ecology International Symposium
Publication Type: Abstract Only
Publication Acceptance Date: August 27, 2004
Publication Date: August 27, 2004
Citation: Whitehead, T.R., Cook, K.L., Cotta, M.A. 2004. Identification of sulfate-reducing bacteria from stored swine manure by pure culture isolation and direct dsra pcr [abstract]. Microbial Ecology International Symposium. Paper No. 228. Technical Abstract: Hydrogen sulfide is a product of the metabolism of sulfate-reducing bacteria (SRB). Hydrogen sulfide is also a component of odor from storage of swine manure from large-scale swine facilities. This odor is a result of microbiological activity during storage, which can be a nuisance to local populace and may contribute to health problems for swine facility workers and animals. However, little is known about the population of SRB in stored swine manure. Swine feces and stored swine manure from a local facility was used as a potential source of SRB. Serial dilutions from the feces and manure using Postgate's medium was used to enrich for SRB, then pure cultures were isolated by streaking on Postgate-agar medium. Total DNA was isolated from the fecal suspension, swine manure slurry, enrichment cultures, and isolated pure cultures for use in direct PCR of the dissimilitory sulfite reductase A (dsrA) gene for phylogenetic analyses, as well as 16S rDNA sequence analyses of pure cultures. Pure culture isolates from feces and manure were presumptively identified as Desulfovibrio species by both 16S rDNA and dsrA sequence analyses. However, direct PCR and sequencing of the dsrA gene from total DNA isolated from the manure slurry also indicated the presence of Desulfobulbus-like SRB as well as Desulfovibiro-like SRB. These results suggest that molecular methods could be developed to monitor different populations of SRB in manure following alterations in swine diet as well as variations in manure handling operations.