Evaluation of modified BACTEC 12B radiometric medium and solid media for culture of Mycobacterium avium subsp. paratuberculosis from sheep

Definitive diagnosis of Johne's disease in ruminants depends on confirming the presence of the causative bacterium, Mycobacterium avium subsp, paratub erculosis, in tissues of the host. This is readily achieved in most ruminan t species by culture. However, culture of clinical specimens from sheep in many countries has been unrewarding. Such a culture from sheep was achieved recently in Australia by using a radiometric culture medium. The aims of t he present study were to evaluate the culture of M. avium subsp, paratuberc ulosis from sheep by using modified BACTEC 12B radiometric medium, to deter mine the sensitivity of culture in relation to histopathology, and to evalu ate a range of solid media. Culture of M. avium subsp. paratuberculosis fro m sheep with Johne's disease is a sensitive method of diagnosis: intestinal tissues from all 43 animals with multibacillary disease and all 22 animals with paucibacillary disease were culture positive, while 98% of feces from 53 animals with multibacillary disease and 48% of feces from 31 animals wi th paucibacillary disease were culture positive. Of sheep without histologi cal evidence of Johne's disease from infected flocks, intestinal tissue fro m 32% of 41 were culture positive, while feces from 17% of 41 were culture positive. Consequently, culture is recommended as the "gold standard" test for detection of ovine Johne's disease. Of the wide range of solid media th at were evaluated, only modified Middlebrook 7H10 and 7H11 agars, which wer e very similar in composition to modified BACTEC 12B medium, yielded growth of ovine strains of M. avium subsp, paratuberculosis, The sensitivity of d etection of M. avium subsp, paratuberculosis on solid media was slightly lo wer than that in modified BACTEC 12B radiometric medium. Both egg yolk and mycobactin J were essential additives for growth of ovine strains of M. avi um subsp, paratuberculosis in both liquid and solid media.