title: Pooled faecal culture for the detection of Mycobacterium avium subsp paratuberculosis in goats.
abstract: OBJECTIVE
  To evaluate pooled faecal culture for herd diagnosis of caprine Johne's disease and relate these findings to faecal shedding rates of Mycobacterium avium subsp paratuberculosis (Map).
  
  
  METHODS
  Radiometric broth culture was applied to several pooling dilutions, and shedding rates were estimated from a regression equation based on bacterial growth rates and known processing losses during radiometric culture.
  
  
  METHODS
  Sixteen faecal samples from goats naturally infected with sheep (n = 3) or cattle (n = 13) strains of Map, were diluted in normal goat faeces from 1 in 5 to 1 in 50. Cultures were confirmed by IS900 polymerase chain reaction and restriction endonuclease analysis, and mycobactin dependency. The numbers of viable Map in the culture inocula were determined by endpoint titration (most probable number) of nine samples and related to a cumulative growth index.
  
  
  RESULTS
  A pooling dilution of 1 in 25 with an incubation period of 10 weeks detected 13 of 16 culture positive goats, all shedding > or = 2 x 10(4) Map per gram of faeces. Two samples containing very low numbers of Map (< 2 x 10(3)/g) were only culture positive from undiluted faeces. Thirteen of 16 goats were considered to be shedding low to moderate concentrations of Map (< 2 x 10(5)/g faeces).
  
  
  CONCLUSIONS
  These data support a pooling dilution of 1 in 25 for application of pooled faecal culture as a diagnostic tool in caprine Johne's disease control. A test based on this dilution would reduce laboratory costs of whole herd testing in goats by approximately 40% relative to serology and 75 to 90% relative to individual faecal culture.
authors.affiliation: New South Wales Department of Primary Industries, Elizabeth Macarthur Agricultural Institute, Camden, NSW 2570. graeme.eamens@dpi.nsw.gov.au
authors.forename: G J
authors.initials: GJ
authors.lastname: Eamens
authors.forename: D M
authors.initials: DM
authors.lastname: Walker
authors.forename: N S
authors.initials: NS
authors.lastname: Porter
authors.forename: S A
authors.initials: SA
authors.lastname: Fell
mesh_terms: D000818:Animals; D015169:Colony Count, Microbial / Q000662:veterinary; D005243:Feces / Q000382:microbiology*; D015511:Goat Diseases / Q000175:diagnosis*; D006041:Goats; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D010080:Oxazoles / Q000032:analysis; D010283:Paratuberculosis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D011874:Radiometry / Q000379:methods / Q000662:veterinary; D015183:Restriction Mapping / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity
chemical_list: D010080:Oxazoles; C018608:mycobactins