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Abstract
Ammonia production in poultry houses has serious implications for flock health and
performance, nutrient value of poultry litter, and energy costs for running poultry
operations. In poultry litter, the conversion of organic N (uric acid and urea) to
NH(4)-N is a microbially mediated process. The urease enzyme is responsible for the
final step in the conversion of urea to NH(4)-N. Cloning and analysis of 168 urease
sequences from extracted genomic DNA from poultry litter samples revealed the presence
of a novel, dominant group of ureolytic microbes (representing 90% of the urease clone
library). Specific primers and a probe were designed to target this novel poultry
litter urease producer (PLUP) group, and a new quantitative real-time PCR assay was
developed. The assay allowed for the detection of 10(2) copies of target urease sequences
per PCR reaction (approximately 1 x 10(4) cells per gram of poultry litter), and the
reaction was linear over 8 orders of magnitude. Our PLUP group was present only in
poultry litter and was not present in environmental samples from diverse agricultural
settings. This novel PLUP group represented between 0.1 to 3.1% of the total microbial
populations (6.0 x 10(6) to 2.4 x 10(8) PLUP cells per gram of litter) from diverse
poultry litter types. The PLUP cell concentrations were directly correlated to the
total cell concentrations in the poultry litter and were found to be influenced by
the physical parameters of the litters (bedding material, moisture content, pH), as
well as the NH(4)-N content of the litters, based on principal component analysis.
Chemical parameters (organic N, total N, total C) were not found to be influential
in the concentrations of our PLUP group in the diverse poultry litters Future applications
of this assay could include determining the efficacy of current NH(4)-N-reducing litter
amendments or in designing more efficient treatment protocols.