title: The prevalence of disseminated Mycobacterium avium subsp. paratuberculosis infection in tissues of healthy ewes from a New Zealand farm with Johne's disease present.
abstract: OBJECTIVE
  To determine the prevalence of disseminated Mycobacterium avium subsp. paratuberculosis (Map) infection in healthy ewes in a flock with a history of clinical Johne's disease.
  
  
  METHODS
  Twenty-four healthy ewes, from a large sheep and cattle farm with a history of clinical Johne's disease in the ewe flock, were randomly selected, euthanased, blood sampled, and examined at necropsy. BACTEC™ radiometric culture for Map was performed on samples of faeces, ileum, mesenteric lymph node, biceps femoris muscle and mononuclear cells in peripheral blood. Serum antibody ELISA tests were performed. Histological sections and Ziehl Neelsen (ZN) stains of impression smears of ileum and mesenteric lymph node were examined for pathological lesions characteristic of Johne's disease and acid fast organisms (AFO). Indirect quantification of Map was performed, using BACTEC radiometric growth indices measuring the time taken for the production of (14)CO(2.)
  
  
  RESULTS
  No histological evidence of Johne's disease or AFO was found in the ileum and mesenteric lymph nodes. Twelve of the 24 ewes (50%) had Map cultured from the ileum (n=6) and/or mesenteric lymph nodes (n=8) while none had Map cultured from the faeces, biceps femoris muscle or blood mononuclear cells. One of the 12 Map culture positive ewes was serum ELISA positive. The culture growth rates in liquid medium suggest low numbers of Map were present in the tissues of the culture positive ewes.
  
  
  CONCLUSIONS
  Fifty per cent of clinically healthy ewes exposed to Map within a Johne's infected flock were Map culture positive in the ileum and/or mesenteric lymph node(s), while the ELISA was positive in 8% of those animals (n=1). There was no faecal shedding of Map and no Map was cultured from skeletal muscle or from blood mononuclear cells suggesting that systemic Map infection, defined as positive culture of Map from skeletal muscle and/or blood, may be uncommon in healthy mixed age ewes without clinical Johne's disease.
  
  
  CONCLUSIONS
  ELISA serology detected 1 of 12 ewes infected with Map whilst none were detected from faecal BACTEC radiometric culture, suggesting biosecurity measures used to control the spread of Map may be of limited use. Map was not cultured from blood mononuclear cells or skeletal muscle, indicating that meat from healthy ewes, from farms where Johne's disease is present, is an unlikely source of Map exposure for humans. Further research is warranted to establish the prevalence and dissemination of Map in tissues outside the alimentary tract of healthy ewes from farms throughout New Zealand where Map is present.
authors.affiliation: Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Palmerston North, New Zealand. s.l.smith@massey.ac.nz
authors.forename: S L
authors.initials: SL
authors.lastname: Smith
authors.forename: D M
authors.initials: DM
authors.lastname: West
authors.forename: P R
authors.initials: PR
authors.lastname: Wilson
authors.forename: G W
authors.initials: GW
authors.lastname: de Lisle
authors.forename: M G
authors.initials: MG
authors.lastname: Collett
authors.forename: C
authors.initials: C
authors.lastname: Heuer
authors.forename: J P
authors.initials: JP
authors.lastname: Chambers
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques; D005260:Female; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D009520:New Zealand / Q000453:epidemiology; D010283:Paratuberculosis / Q000453:epidemiology*; D015995:Prevalence; D012756:Sheep; D012757:Sheep Diseases / Q000453:epidemiology / Q000382:microbiology*