Immunomagnetic recovery of Mycobacterium bovis from naturally infected environmental samples

• Published in: Letters in applied microbiology Vol. 43; no. 4; pp. 364 - 369
• Main Authors: Sweeney, F.P, Courtenay, O, Ul-Hassan, A, Hibberd, V, Reilly, L.A, Wellington, E.M.H
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01.10.2006; Blackwell Publishing Ltd; Blackwell Science
• Subjects: Animals; Bacteriological Techniques - methods; badger; Biological and medical sciences; bovine tuberculosis; Cattle; Colony Count, Microbial - methods; environment; Feces - microbiology; Fundamental and applied biological sciences. Psychology; immunomagnetic capture; Immunomagnetic Separation - methods; Meles meles; Microbiology; Mycobacterium bovis; Mycobacterium bovis - genetics; Mycobacterium bovis - growth & development; Mycobacterium bovis - isolation & purification; Polymerase Chain Reaction; Soil Microbiology; Immunomagnetic method; Applied microbiology; Mycobacterial infection; Recovery; Bacteria; Ungulata; Fissipedia; Carnivora; bovine tuberculosis; Bovine; Mycobacterium bovis; Sample; badger; Infection; Vertebrata; Mammalia; Tuberculosis; Meles meles; Mycobacteriales; immunomagnetic capture; Bacteriosis; Mycobacteriaceae; Environment; Actinomycetes; Artiodactyla
• ISSN: 0266-8254; 1472-765X; 1365-2673
• DOI: 10.1111/j.1472-765X.2006.01983.x

To adapt an immunomagnetic capture (IMC) technique to concentrate and cultivate Mycobacterium bovis from environmental samples including soil, faeces and urine. 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. The IMC techniques can be applied to isolate viable wild type Myco. bovis from naturally contaminated environmental samples. 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.