The objective of the present study was to evaluate the suitability of environmental sampling to screen Czech dairy herds to detect Mycobacterium avium ssp. paratuberculosis (MAP) and to find the most convenient location for the MAP detection in the lactating cow area. Environmental samples (ES, n = 72) from milking parlour holding pens (n = 19), milking alleyways (n = 19) and free-stall alleyways (n = 34) from 19 herds were simultaneously tested to detect MAP by a quantitative PCR (qPCR) and bacterial culture. Eight and thirteen samples from the milking parlour holding pens, twelve and eleven samples from the milking alleyways and eleven and eighteen samples from the free-stall alleyways were qPCR and culture positive, respectively. A 4.6 times higher probability of being culture positive than qPCR positive was detected for the assessable MAP detection results from the free-stall alleyways [P = 0.008 6, odds ration (OR) = 4.572 8)] and no association was found between the results from the milking parlour holding pens (P = 0.191 4) and the milking alleyways (P > 0.999 9) and the diagnostic method used. The percentage of qPCR-positive samples in the tested locations was detected for the milking alleyways (63.2%), free-stall alleyways and milking parlour holding pens. The herd infectious status was in agreement with 16 (84.2%), 14 (73.7%) and 12 (63.2%) qPCR results from the milking alleyways, free-stall alleyways (32.4%) and milking parlour holding pens (42.1%), respectively. No statistically significant differences were detected for these results (P = 0.396 1). MAP was detected by the qPCR and bacterial culture in all three locations where the ES were collected. We suggest an environmental sampling followed by MAP detection by qPCR as an easy-to-perform time-saving protocol for MAP screening in Czech dairy herds. Although the milking alleyways seem to be the most convenient location for the environmental sampling, this assumption was not statistically supported.

• Klíčová slova
• Johne’s disease, faecal culture, lactating cow, quantitative PCR, sampling location,
• Publikační typ
• časopisecké články MeSH

The aim of this study was to monitor the persistence of Mycobacterium avium subsp. paratuberculosis in environmental samples taken from a Holstein farm with a long history of clinical paratuberculosis. A herd of 606 head was eradicated, and mechanical cleaning and disinfection with chloramine B with ammonium (4%) was carried out on the farm; in the surrounding areas (on the field and field midden) lime was applied. Environmental samples were collected before and over a period of 24 months after destocking. Only one sample out of 48 (2%) examined on the farm (originating from a waste pit and collected before destocking) was positive for M. avium subsp. paratuberculosis by cultivation on solid medium (Herrold's egg yolk medium). The results using real-time quantitative PCR (qPCR) showed that a total of 81% of environmental samples with an average mean M. avium subsp. paratuberculosis cell number of 3.09 × 10(3) were positive for M. avium subsp. paratuberculosis before destocking compared to 43% with an average mean M. avium subsp. paratuberculosis cell number of 5.86 × 10(2) after 24 months. M. avium subsp. paratuberculosis-positive samples were detected in the cattle barn as well as in the calf barn and surrounding areas. M. avium subsp. paratuberculosis was detected from different matrices: floor and instrument scrapings, sediment, or scraping from watering troughs, waste pits, and cobwebs. M. avium subsp. paratuberculosis DNA was also detected in soil and plants collected on the field midden and the field 24 months after destocking. Although the proportion of positive samples decreased from 64% to 23% over time, the numbers of M. avium subsp. paratuberculosis cells were comparable.

• MeSH
• bakteriologické techniky metody   MeSH
• bydlení zvířat MeSH
• DNA bakterií genetika   MeSH
• hospodářská zvířata MeSH
• kontrola infekce metody   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• mikrobiologie životního prostředí * MeSH
• Mycobacterium avium subsp. paratuberculosis genetika   růst a vývoj   izolace a purifikace   MeSH
• nemoci skotu diagnóza   mikrobiologie   MeSH
• paratuberkulóza diagnóza   mikrobiologie   MeSH
• rostliny mikrobiologie   MeSH
• skot MeSH
• transpozibilní elementy DNA MeSH
• veterinární lékařství metody   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• transpozibilní elementy DNA MeSH

The aim of this study was to demonstrate the persistence of Mycobacterium avium subsp. paratuberculosis (MAP) in soil and colonization of different plant parts after deliberate exposure to mouflon feces naturally contaminated with different amounts of MAP. Samples of aerial parts of plants, their roots, and the soil below the roots were collected after 15 weeks and examined using IS900 real-time quantitative PCR (qPCR) and cultivation. Although the presence of viable MAP cells was not demonstrated, almost all samples were found to be positive using qPCR. MAP IS900 was not only found in the upper green parts, but also in the roots and soil samples (from 1.00 × 10(0) to 6.43 × 10(3)). The level of soil and plant contamination was influenced mainly by moisture, clay content, and the depth from which the samples were collected, rather than by the initial concentration of MAP in the feces at the beginning of the experiment.

• MeSH
• feces mikrobiologie   MeSH
• Mycobacterium avium subsp. paratuberculosis genetika   izolace a purifikace   MeSH
• nemoci ovcí mikrobiologie   MeSH
• ovce domácí * MeSH
• paratuberkulóza mikrobiologie   MeSH
• půda chemie   MeSH
• půdní mikrobiologie * MeSH
• rostliny mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• půda MeSH