Register      Login
Microbiology Australia Microbiology Australia Society
Microbiology Australia, bringing Microbiologists together
RESEARCH ARTICLE

Relative abundance of Mycobacterium in ovine Johne’s disease

Andy O Leu A , Paul Pavli B C , David M Gordon A , Jeff Cave D , Jacek M Gowzdz E , Nick Linden D , Grant Rawlin E , Gwen E Allison A C and Claire L O’Brien A B C F
+ Author Affiliations
- Author Affiliations

A Research School of Biology, Australian National University, Acton, ACT 0200, Australia

B Gastroenterology and Hepatology Unit, Canberra Hospital, Garran, ACT 2605, Australia

C Medical School, Australian National University, Acton, ACT 0200, Australia

D Department of Environment and Primary Industries, Wodonga, Vic. 3690, Australia

E AgriBio, Bundoora, Vic. 3083, Australia

F Corresponding author. Tel: +61 2 6244 4023, Email: claire.obrien@anu.edu.au

Microbiology Australia 36(1) 32-36 https://doi.org/10.1071/MA15010
Published: 6 March 2015

Abstract

No study has determined what proportion of the total microbiota comprises the genus Mycobacterium in ovine Johne's disease (OJD) tissues. We aimed to assess the relative abundance of Mycobacterium in the ileocaecal lymph node, involved and uninvolved ileal mucosa from sheep with and without OJD, using three extraction methods. Eight sheep, four with and four without OJD, were recruited. Pyrosequencing of the 16Sr RNA gene amplicons for all samples revealed that Mycobacterium represented between 0-92% (average 38%) of the total microbiota of samples from sheep with OJD, and 0-85% (average 13%) of sheep without OJD. Only sheep with OJD had samples that were positive for the IS900 (MAP) element. Mycobacterial strains other than MAP may provide competitive exclusion of MAP and should be further investigated.


References

[1]  Salem, M. et al. (2013) Mycobacterium avium subspecies paratuberculosis: an insidious problem for the ruminant industry. Trop. Anim. Health Prod. 45, 351–366.
Mycobacterium avium subspecies paratuberculosis: an insidious problem for the ruminant industry.Crossref | GoogleScholarGoogle Scholar | 23054804PubMed |

[2]  Cheung, M.K. et al. (2013) Sputum microbiota in tuberculosis as revealed by 16S rRNA pyrosequencing. PLoS ONE 8, e54574.
Sputum microbiota in tuberculosis as revealed by 16S rRNA pyrosequencing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXit1ChsLg%3D&md5=c24867699af12fecab1bae29f719b120CAS | 23365674PubMed |

[3]  O’Brien, C.L. et al. (2014) Detection of bacterial DNA in lymph nodes of Crohn’s disease patients using high throughput sequencing. Gut 63, 1596–1606.
Detection of bacterial DNA in lymph nodes of Crohn’s disease patients using high throughput sequencing.Crossref | GoogleScholarGoogle Scholar | 24429583PubMed |

[4]  Park, K.T. et al. (2014) Development of a novel DNA extraction method for identification and quantification of Mycobacterium avium subsp. paratuberculosis from tissue samples by real-time PCR. J. Microbiol. Methods 99, 58–65.
Development of a novel DNA extraction method for identification and quantification of Mycobacterium avium subsp. paratuberculosis from tissue samples by real-time PCR.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXks1Wgtrs%3D&md5=7b1ece82390397de1004aa8abd2bafafCAS | 24534783PubMed |

[5]  Logar, K. et al. (2012) Evaluation of combined high-efficiency DNA extraction and real-time PCR for detection of Mycobacterium avium subsp. paratuberculosis in subclinically infected dairy cattle: comparison with faecal culture, milk real-time PCR and milk ELISA. BMC Vet. Res. 8, 49.
Evaluation of combined high-efficiency DNA extraction and real-time PCR for detection of Mycobacterium avium subsp. paratuberculosis in subclinically infected dairy cattle: comparison with faecal culture, milk real-time PCR and milk ELISA.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhsVaktLnI&md5=746caea40c02dbbf19c742214ea008efCAS | 22551054PubMed |

[6]  Schloss, P.D. et al. (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl. Environ. Microbiol. 75, 7537–7541.
Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXis1yltw%3D%3D&md5=2bd3a329be90d06651a79c07a2341d58CAS | 19801464PubMed |

[7]  Gwozdz, J.M. et al. (2000) Vaccination against paratuberculosis of lambs already infected experimentally with Mycobacterium avium subspecies paratuberculosis. Aust. Vet. J. 78, 560–566.
Vaccination against paratuberculosis of lambs already infected experimentally with Mycobacterium avium subspecies paratuberculosis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3cvmt1Smtg%3D%3D&md5=c549ce8a8a7c6aaf7b3119603567f7e9CAS | 10979513PubMed |

[8]  Whittington, R.J. et al. (2013) Development and validation of a liquid medium (M7H9C) for routine culture of Mycobacterium avium subsp. paratuberculosis to replace modified Bactec 12B medium. J. Clin. Microbiol. 51, 3993–4000.
Development and validation of a liquid medium (M7H9C) for routine culture of Mycobacterium avium subsp. paratuberculosis to replace modified Bactec 12B medium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXjtlWrsbs%3D&md5=1bf4eff20334a79f64159b42f83efbabCAS | 24048541PubMed |

[9]  Sting, R. et al. (2014) Detection of Mycobacterium avium subsp. paratuberculosis in faeces using different procedures of pre-treatment for real-time PCR in comparison to culture. Vet. J. 199, 138–142.
Detection of Mycobacterium avium subsp. paratuberculosis in faeces using different procedures of pre-treatment for real-time PCR in comparison to culture.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhvVKisbvP&md5=b6af46cbb4d2b2b4861831a39c009e7fCAS | 24280588PubMed |

[10]  Pryde, S.E. et al. (2002) The microbiology of butyrate formation in the human colon. FEMS Microbiol. Lett. 217, 133–139.
The microbiology of butyrate formation in the human colon.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XptlCqs7g%3D&md5=22bb89ffcdf2b3809b3cb830f1d386eaCAS | 12480096PubMed |