title: Identification of a Brevibacterium marker gene specific to poultry litter and development of a quantitative PCR assay.
abstract: OBJECTIVE
  To identify a DNA sequence specific to a bacterium found in poultry litter that was indicative of faecal contamination by poultry sources.
  
  
  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 10(7) -10(9) gene copies per gram in soiled litter, up to 10(5) gene copies per gram in spread-site soils, and 10(7) gene copies per litre in field run-off water. Results were corroborated by a blinded study conducted by a second laboratory.
  
  
  CONCLUSIONS
  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.
  
  
  CONCLUSIONS
  This study describes the first quantitative, sensitive and specific microbial source tracking method for the detection of poultry litter contamination.
authors.affiliation: North Wind, Inc., Idaho Falls, ID, USA.
authors.forename: J L
authors.initials: JL
authors.lastname: Weidhaas
authors.forename: T W
authors.initials: TW
authors.lastname: Macbeth
authors.forename: R L
authors.initials: RL
authors.lastname: Olsen
authors.forename: M J
authors.initials: MJ
authors.lastname: Sadowsky
authors.forename: D
authors.initials: D
authors.lastname: Norat
authors.forename: V J
authors.initials: VJ
authors.lastname: Harwood
mesh_terms: D000818:Animals; D001951:Brevibacterium / Q000145:classification / Q000235:genetics*; D002417:Cattle / Q000382:microbiology; D002645:Chickens / Q000382:microbiology*; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods; D005243:Feces / Q000382:microbiology*; D005798:Genes, Bacterial; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D011200:Poultry / Q000382:microbiology; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012722:Sewage / Q000382:microbiology; D012987:Soil / Q000032:analysis; D012988:Soil Microbiology; D013552:Swine / Q000382:microbiology; D014481:United States
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012722:Sewage; D012987:Soil