Factors influencing the isolation of Mycobacterium avium subsp paratuberculosis from bovine fecal samples

A modified procedure was used for culture of Mycobacterium paratuberculosis (Mptb) from bovine feces. Bovine fecal samples were decontaminated with Na OH, exposed to a mixture of oxalic acid and malachite green, incubated in a mixture of neomycin and amphotericin B. Decontaminated specimens were inoc ulated onto modified Lowenstein-Jensen medium. Specimens processed by high- speed centrifugation showed growth earlier than specimens prepared by low s peed centrifugation. However, the overall number of positive cultures at 16 weeks was not different for the 2 methods. When infected dairy herds were sampled 4 times at 6-month intervals and culture-positive cows were culled, the prevalence of infected cattle declined over time. After selective cull ing, the cattle left in the herds shed low numbers of Mptb, which explains why it took longer for cultures to become positive. No heifers younger than 11 months were culture positive, but heifers 13-14 months of age were more frequently culture positive than were heifers of any other age. The 16-wee k culture period is needed with this method to detect cattle shedding low n umbers of Mptb. High-speed centrifugation of samples does not increase the efficiency of identification of animals shedding Mptb.