AIMS: To adapt an immunomagnetic capture (IMC) technique to concentrate and cultivate Mycobacterium bovis from environmental samples including soil, faeces and urine. METHODS AND 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. SIGNIFICANCE AND IMPACT OF STUDY: 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.
AIMS: To adapt an immunomagnetic capture (IMC) technique to concentrate and cultivate Mycobacterium bovis from environmental samples including soil, faeces and urine. METHODS AND 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. SIGNIFICANCE AND IMPACT OF STUDY: 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 bovinetuberculosis persistence.
Authors: Linda D Stewart; James McNair; Lyanne McCallan; Suzan Thompson; Leonid A Kulakov; Irene R Grant Journal: J Clin Microbiol Date: 2012-02-08 Impact factor: 5.948
Authors: F P Sweeney; O Courtenay; V Hibberd; R G Hewinson; L A Reilly; W H Gaze; E M H Wellington Journal: Appl Environ Microbiol Date: 2007-09-28 Impact factor: 4.792
Authors: Joanne L Hardstaff; Mark T Bulling; Glenn Marion; Michael R Hutchings; Piran C L White Journal: BMC Vet Res Date: 2012-06-27 Impact factor: 2.741
Authors: Alessandra Pontiroli; Emma Rachel Travis; Francis Patrick Sweeney; David Porter; William Hugo Gaze; Sam Mason; Victoria Hibberd; Jennifer Holden; Orin Courtenay; Elizabeth Margaret Helen Wellington Journal: PLoS One Date: 2011-03-23 Impact factor: 3.240
Authors: Emma R Travis; William H Gaze; Alessandra Pontiroli; Francis P Sweeney; David Porter; Sam Mason; Matthew J C Keeling; Rebecca M Jones; Jason Sawyer; Alicia Aranaz; Elena Castellanos Rizaldos; Jennifer Cork; Richard J Delahay; Gavin J Wilson; R Glyn Hewinson; Orin Courtenay; Elizabeth M H Wellington Journal: PLoS One Date: 2011-11-14 Impact factor: 3.240