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Comparison of different sampling sites and techniques for the detection of Mycobacterium avium subsp. paratuberculosis in environmental fecal samples in paratuberculosis positive cattle herds
Khol, J.L.; Mattes, M.; Dünser, M...
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Objectives: Mycobacterium avium subsp. paratuberculosis (MAP) is causing paratuberculosis (Johne ́s disease, JD) in cattle and is known to survive for an extended period of time in the environment. The objective of this study was, to evaluate in which areas within a barn MAP can be detected in positive cattle farms and to compare different sampling sites. Thereby, potential areas for MAP transmission, as well as the most promising places for the detection of MAP in positive cattle premises, should be identified.
Material and Methods: During the study, 14 Austrian dairy and beef operations were tested for the presence of MAP twice in a six months interval. In each farm at least one confirmed clinical case of JD was detected within a year prior to the study and the herd was therefore considered to be MAP positive.
On every farm, 7-10 paired environmental fecal samples from specific sites were taken. Sampling locations depended on the type of operation and included the calving area, alleyways, equipment, milking parlor and manure storage sites. Fecal samples were tested for MAP both by bacteriological culture on Herrold ́s Egg Yolk Medium and real time PCR for IS900 (Adiagene, Saint Brieuc, France), at the Austrian Na- tional Reference Laboratory for paratuberculosis.
Results: All farms enrolled in the study had at least one positive environmental fecal sample, confirming the classification as MAP positive. Fecal samples collected from the slurry pit, the alleyways in the feeding area as well as the manure channels (tie stall barns) proved to be most likely MAP positive. Altogether, 42.3% of the samples from the slurry pit were positive by culture and 51.9% by PCR, samples from the alleyways from the feeding area showed 44.4% and 30.0% positive results, and in the manure channel 87.5% of the collected samples were MAP positive by culture an 50.0% by PCR, respectively. The sensitivity of the samples could be increased significantly by collecting two samples from each site and reached 100% at the herd level, when several sample sites were combined within a farm.
Conclusions: The results of the present study indicate, that manure storage sites, as well as the highly frequented alleyways in a barn seem to be the most promising sites for the detection of MAP by environmental fecal samples. Based on these results, the use of environmental fecal samples seems to be a useful tool to assess the MAP herd level in cattle.
Keywords: Paratuberculosis, Johne ́s disease, Mycobacterium avium subsp. paratuberculosis, environmental fecal sampling.
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Affiliation of the authors at the time of publication
University Clinic for Ruminants, University of Veterinary Medicine Vienna, Vienna, Austria;
Austrian Agency for Health and Food Safety, Linz, Austria.
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