Survival of Mycobacterium avium subsp. paratuberculosis in biofilms on livestock watering trough materials.

Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of Johne's disease, a chronic enteric infection that affects ruminants. Despite the ubiquitous occurrence of Mycobacterium sp. in nature and the fact that Johne's disease has been reported worldwide, little research has been done to assess its survival in agricultural environments. The goal of this 365-day study was to evaluate the ability of Map to persist in mixed-community biofilms on materials commonly used to construct livestock watering troughs. Map was inoculated into 32l of trough water containing either concrete, plastic, galvanized or stainless steel trough materials. The concentration of Map was determined by using quantitative, real-time PCR to target the IS900 sequence in DNA extracts. High concentrations of Map were detected on all trough materials after 3 days (around 1 x 10(5)cells cm(-2)). Based on the best-fit slopes, the time required for a 99% reduction (t(99)) in biofilm-associated Map cells was 144 and 115 days for plastic and stainless steel trough materials, respectively. Map concentrations did not decrease on concrete and galvanized steel trough materials. These results suggest that Map survives well in biofilms present on livestock watering trough materials. To inhibit spread of this organism and exposure of susceptible animals to Map on infected farms, best management practices aimed at maintaining biofilm-free trough surfaces should be included in any Johne's control plan.