title: Immunomagnetic recovery of Mycobacterium bovis from naturally infected environmental samples.
abstract: OBJECTIVE
  To adapt an immunomagnetic capture (IMC) technique to concentrate and cultivate Mycobacterium bovis from environmental samples including soil, faeces and urine.
  
  
  RESULTS
  Cells of Myco. bovis BCG and wild-type Myco. bovis were successfully isolated and cultured from seeded and naturally infected materials respectively. The IMC cell recovery estimated by colony forming units (CFUs) counts ranged from 0.10% to 0.16% for spiked media, and 0.15-0.36% for naturally infected soil and faeces. Recovery estimated by cell counts calculated using semi-quantitative PCR ranged from 80.3% to 88.6% for spiked and 84.1-88.2% for naturally infected material. The differences in the recovery rates estimated by CFUs compared with pixel intensity is likely to be due to clustering of cells on culture plates, thereby underestimating the true cell count.
  
  
  CONCLUSIONS
  The IMC techniques can be applied to isolate viable wild type Myco. bovis from naturally contaminated environmental samples.
  
  
  CONCLUSIONS
  Cultivation of Myco. bovis from environmental samples using traditional methods is extremely problematic. Here, we demonstrate a novel development of IMC techniques that will greatly facilitate the study of the organism in situ in order to assess its epidemiological importance in bovine tuberculosis persistence.
authors.affiliation: Department of Biological Sciences, University of Warwick, Coventry, UK. f.p.sweeney@warwick.ac.uk
authors.forename: F P
authors.initials: FP
authors.lastname: Sweeney
authors.forename: O
authors.initials: O
authors.lastname: Courtenay
authors.forename: A
authors.initials: A
authors.lastname: Ul-Hassan
authors.forename: V
authors.initials: V
authors.lastname: Hibberd
authors.forename: L A
authors.initials: LA
authors.lastname: Reilly
authors.forename: E M H
authors.initials: EM
authors.lastname: Wellington
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D002417:Cattle; D015169:Colony Count, Microbial / Q000379:methods; D005243:Feces / Q000382:microbiology*; D018189:Immunomagnetic Separation / Q000379:methods; D009163:Mycobacterium bovis / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012988:Soil Microbiology