Identification of a Brevibacterium marker gene specific to poultry litter and development of a quantitative PCR assay |
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Authors: | JL Weidhaas TW Macbeth RL Olsen MJ Sadowsky D Norat VJ Harwood |
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Institution: | 1. North Wind, Inc., Idaho Falls, ID, USA;2. CDM, Denver, CO, USA;3. Department of Soil, Water, and Climate and BioTechnology Institute, University of Minnesota, St. Paul, MN, USA;4. Department of Biology, University of South Florida, Tampa, FL, USA |
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Abstract: | Aim: To identify a DNA sequence specific to a bacterium found in poultry litter that was indicative of faecal contamination by poultry sources. Methods and Results: Faecally contaminated poultry litter and soils were used as source material for the development of a quantitative polymerase chain reaction (qPCR) method targeting the 16S rRNA gene of a Brevibacterium sp. The identified sequence had 98% nucleotide identity to the 16S rRNA gene of Brevibacterium avium. The qPCR method was tested on 17 soiled litter samples; 40 chicken faecal samples; and 116 nontarget faecal samples from cattle, swine, ducks, geese, and human sewage collected across the United States. The 571‐bp product was detected in 76% of poultry‐associated samples, but not in 93% of faecal samples from other sources. Marker concentrations were 107–109 gene copies per gram in soiled litter, up to 105 gene copies per gram in spread‐site soils, and 107 gene copies per litre in field run‐off water. Results were corroborated by a blinded study conducted by a second laboratory. Conclusion: The poultry‐specific PCR product is a useful marker gene for assessing the impact of faecal contamination as a result of land‐applied poultry litter. Significance and Impact of the Study: This study describes the first quantitative, sensitive and specific microbial source tracking method for the detection of poultry litter contamination. |
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Keywords: | Brevibacterium faecal contamination microbial source tracking poultry qPCR waste disposal water quality |
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