Detection of Ammonia-Oxidizing Beta-Proteobacteria in Swine Waste Treatment Systems.

Abstract

In order to obtain supporting evidence for biological denitrification in anaerobic lagoons degenerate β-Proteobacterial AOB primers were used to create and sequence clone libraries to detect the presence of ammonia-oxidizing bacteria at three field sites. Although there were PCR products from almost all samples, the clone libraries that were created show that not all PCR reactions produce only PCR products from ammonia-oxidizing bacteria. However, these primers did verify the presence of ammonia-oxidizers at one site, although their presence was not verified at the other sites. The presence of ammonia-oxidizers at the Battelle site implies that aerobic ammonia-oxidation is occurring. Clones were created and sequenced that were significantly different from other known sequences and tended to form very closely related phylogenetic groups. These phylogenetic groups were not isolated to one field site, and often more than one site had representatives in a closely related group. Future research in this field includes the design of new primer sets based on the sequences of the nitrifying bacteria clones reported in this research, creation of enrichment cultures, and use of new primers for fluorescent in situ hybridization.