Molecular Epidemiology of Mycobacterium avium subsp.paratuberculosis: IS900 Restriction Fragment Length Polymorphism and IS1311 Polymorphism Analyses of Isolates from Animals and a Human in Australia

Abstract

The distribution and prevalence of strains of Mycobacterium avium subsp. paratuberculosis were determined among sheep, cattle, and other species with Johne's disease in Australia. A total of 328 isolates were evaluated from numerous farms in New South Wales, Victoria, Tasmania, and South Australia, Australia. Restriction fragment length polymorphism (RFLP) analysis of genomic DNA usingBstEII and an IS900 probe and IS1311 polymorphism analysis using PCR and restriction endonuclease analysis (PCR-REA) was used to classify isolates as cattle (C) or sheep (S) strains. IS1311 PCR-REA provided similar information to IS900 RFLP analysis but was more useful than RFLP analysis where DNA was degraded or scarce. Twelve IS900 RFLP types were found. Johne's disease in sheep was always due to S strains, while cattle were infected only with C strains. RFLP type S1 was the dominant strain in sheep in New South Wales (97% of isolates) and was the only strain found in sheep from Victoria. Seven RFLP types were present in cattle. RFLP types C3 and C1 were most common (collectively, 85% of isolates), but C1 was not found in New South Wales and C3 was present in dairy cattle but not in beef cattle in Victoria. These differences may be explained by restricted livestock trading patterns between different segments of the cattle industry. Up to five RFLP types were present in some geographic regions in Victoria, while up to three RFLP types were found among cattle on some farms. Individual cattle usually were infected with only one RFLP type, but one animal was infected with both C5 and CU4. Two isolates from goats were C type as were three from alpacas, one from a rhinoceros, and two from a human with Crohn's disease. The prevalences of specific RFLP types in Australia differ from those reported in Europe and elsewhere. Given the existence of geographical and farm enterprise differences in IS900 RFLP type, this technique may be applied selectively to trace the spread of Johne's disease, at least in the cattle industries. As these observations reflect past exposure of livestock to M. avium subsp.paratuberculosis, the monitoring of strains present in animals in Australia is continuing.

How does a Mycobacterium change its spots? Applying molecular tools to track diverse strains of Mycobacterium avium subspecies paratuberculosis

“…Some pathogenic differences between these lineages have also been noted (Motiwala et al 2006;Gollnick et al 2007;Borrmann et al 2011). Whittington et al (2000Whittington et al ( , 2001 and Marsh et al (1999) developed the IS1311 PCR amplification-restriction enzyme analysis and used it successfully to distinguish C strains from S strains on a genotype level. This technique could also be used to distinguish other subspecies of Myco.…”

Use of Molecular Epidemiology in Veterinary Practice

“…This finding means that sheep do not pose a risk of MAP infection for cattle. An Australian study also differentiated ovine MAP from bovine MAP and put MAP from goats, alpacas, a rhinoceros, and two humans into the same group as bovine MAP [108]. Later studies, however, showed that contact of calves with paratuberculous sheep can result in presence of ovine MAP in cattle.…”

Specificity of absorbed ELISA and agar gel immuno‐diffusion tests for paratuberculosis in goats with observations about use of these tests in infected goats

“…The genotype of the infecting strain of M a paratuberculosis was determined for each case. This was based on the cultural characteristics of isolates of M a paratuberculosis, 9 restriction fragment length polymorphism analysis and/or IS1311 polymorphism analysis, 10,11 or based on epidemiological criteria where the case was detected by tracing back from a property where strain typing had already been conducted. Twelve sera were derived from goats infected with cattle strains of M a paratuberculosis on five farms while seven were derived from goats infected with sheep strains of M a paratuberculosis on three farms.…”