- Z. Dimareli (1)
- K. Dohmann (1)
- S. Englund (1)
- G.F. Gerlach (1)
- A. Greig (1)
- J.F.T. Griffin (1)
- I. Heron (1)
- D. Herthnek (1)
- L. Juan de (1)
- M. Kopecna (2)
- C.G. Mackintosh (1)
- L. May (1)
- R. O'Brien (1)
- I. Pavlik (2)
- J.M. Sharp (1)
- K. Stevenson (1)
- V.C. Thibault (1)
- P.T.J. Willemsen (1)
- P. Willemsen (1)
- R.N. Zadoks (1)
Occurrence of Mycobacterium avium subspecies paratuberculosis across host species and European countries with evidence for transmission between wildlife and domestic ruminants
Stevenson, K. ; Alvarez, J. ; Bakker, D. ; Biet, F. ; Juan, L. de; Denham, S. ; Dimareli, Z. ; Dohmann, K. ; Gerlach, G.F. ; Heron, I. ; Kopecna, M. ; May, L. ; Pavlik, I. ; Sharp, J.M. ; Thibault, V.C. ; Willemsen, P. ; Zadoks, R.N. ; Greig, A. - \ 2009
BMC Microbiology 9 (2009). - ISSN 1471-2180 - 13 p.
fragment-length-polymorphism - genetic diversity - s-strain - molecular characterization - sequence polymorphisms - culture-media - infection - cattle - is900 - rabbits
Background: Mycobacterium avium subspecies paratuberculosis (Map) causes an infectious chronic enteritis (paratuberculosis or Johne's disease) principally of ruminants. The epidemiology of Map is poorly understood, particularly with respect to the role of wildlife reservoirs and the controversial issue of zoonotic potential (Crohn's disease). Genotypic discrimination of Map isolates is pivotal to descriptive epidemiology and resolving these issues. This study was undertaken to determine the genetic diversity of Map, enhance our understanding of the host range and distribution and assess the potential for interspecies transmission. Results: 164 Map isolates from seven European countries representing 19 different host species were genotyped by standardized IS900 - restriction fragment length polymorphism (IS900-RFLP), pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphisms (AFLP) and mycobacterial interspersed repeat unit-variable number tandem repeat (MIRU-VNTR) analyses. Six PstI and 17 BstEII IS900-RFLP, 31 multiplex [SnaBI-SpeI] PFGE profiles and 23 MIRU-VNTR profiles were detected. AFLP gave insufficient discrimination of isolates for meaningful genetic analysis. Point estimates for Simpson's index of diversity calculated for the individual typing techniques were in the range of 0.636 to 0.664 but a combination of all three methods increased the discriminating power to 0.879, sufficient for investigating transmission dynamics. Two predominant strain types were detected across Europe with all three typing techniques. Evidence for interspecies transmission between wildlife and domestic ruminants on the same property was demonstrated in four cases, between wildlife species on the same property in two cases and between different species of domestic livestock on one property. Conclusion: The results of this study showed that it is necessary to use multiple genotyping techniques targeting different sources of genetic variation to obtain the level of discrimination necessary to investigate transmission dynamics and trace the source of Map infections. Furthermore, the combination of genotyping techniques may depend on the geographical location of the population to be tested. Identical genotypes were obtained from Map isolated from different host species co-habiting on the same property strongly suggesting that interspecies transmission occurs. Interspecies transmission of Map between wildlife species and domestic livestock on the same property provides further evidence to support a role for wildlife reservoirs of infection
Sensitive detection of Myobacterium avium subsp paratuberculosis in bovine semen by real-time PCR
Herthnek, D. ; Englund, S. ; Willemsen, P.T.J. ; Bolske, G. - \ 2006
Journal of Applied Microbiology 100 (2006)5. - ISSN 1364-5072 - p. 1095 - 1102.
polymerase-chain-reaction - mycobacterium-paratuberculosis - crohns-disease - johnes-disease - diagnosis - organs - is900 - milk - dna - herpesvirus
Aims: To develop a fast and sensitive protocol for detection of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine semen and to make a critical evaluation of the analytical sensitivity. Methods and Results: Processed semen was spiked with known amounts of MAP. Semen from different bulls as well as semen of different dilutions was tested. The samples were treated with lysing agents and beadbeating and the DNA was extracted with phenol and chloroform. Real-time PCR with a fluorescent probe targeting the insertion element IS900 detected as few as 10 organisms per sample of 100 ¿l semen. PCR-inhibition was monitored by inclusion of an internal control. Pre-treatment with immunomagnetic separation was also evaluated, but was not shown to improve the overall sensitivity. Conclusions: Real-time PCR is a sensitive method for detection of MAP in bovine semen. Lysis by mechanical disruption followed by phenol and chloroform extraction efficiently isolated DNA and removed PCR-inhibitors. Significance and Impact of the Study: The high sensitivity of the applied method allows reliable testing of bovine semen used for artificial insemination to prevent the spread of Johne's disease, caused by MAP.
Immunological and molecular characterization of susceptibility in relationship to bacterial strain differences in Mycobacterium avium subsp paratuberculosis infection in the red deer (Cervus elaphus)
O'Brien, R. ; Mackintosh, C.G. ; Bakker, D. ; Kopecna, M. ; Pavlik, I. ; Griffin, J.F.T. - \ 2006
Infection and Immunity 74 (2006)6. - ISSN 0019-9567 - p. 3530 - 3537.
fragment-length-polymorphism - johnes-disease - protective efficacy - immune-responses - new-zealand - wild ruminants - farmed deer - tuberculosis - sheep - is900
Johne's disease (JD) infection, caused by Mycobacterium avium subsp. paratuberculosis, represents a major disease problem in farmed ruminants. Although JD has been well characterized in cattle and sheep, little is known of the infection dynamics or immunological response in deer. In this study, typing of M. avium subsp. paratuberculosis isolates from intestinal lymphatic tissues from 74 JD-infected animals showed that clinical isolates of M. avium subsp. paratuberculosis from New Zealand farmed red deer were exclusively of the bovine strain genotype. The susceptibility of deer to M. avium subsp. paratuberculosis was further investigated by experimental oral-route infection studies using defined isolates of virulent bovine and ovine M. avium subsp. paratuberculosis strains. Oral inoculation with high (109 CFU/animal) or medium (107 CFU/animal) doses of the bovine strain of M. avium subsp. paratuberculosis established 100% infection rates, compared to 69% infection following inoculation with a medium dose of the ovine strain. The high susceptibility of deer to the bovine strain of M. avium subsp. paratuberculosis was confirmed by a 50% infection rate following experimental inoculation with a low dose of bacteria (103 CFU/animal). This study is the first to report experimental M. avium subsp. paratuberculosis infection in red deer, and it outlines the strong infectivity of bovine-strain M. avium subsp. paratuberculosis isolates for cervines.