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.
Alinovi CA, Ward MP, Lin TL, Moore GE, Wu CC. Real-time PCR, compared to liquid and solid culture media and ELISA, for detection of Mycobacterium avium ssp. paratuberculosis. Vet Microbiol. 2009 Apr 14;136(1-2):177-9. https://doi.org/10.1016/j.vetmic.2008.10.012
Berghaus RD, Farver TB, Anderson RJ, Jaravata CC, Gardner IA. Environmental sampling for detection of Mycobacterium avium ssp. paratuberculosis on large California dairies. J Dairy Sci. 2006 Mar;89(3):963-70. https://doi.org/10.3168/jds.S0022-0302(06)72161-0
Chui LW, King R, Lu P, Manninen K, Sim J. Evaluation of four DNA extraction methods for the detection of Mycobacterium avium subsp. paratuberculosis by polymerase chain reaction. Diagn Microbiol Infect Dis. 2004 Jan;48(1):39-45. https://doi.org/10.1016/j.diagmicrobio.2003.08.007
Corbett CS, Naqvi SA, Bauman CA, De Buck J, Orsel K, Uehlinger F, Kelton DF, Barkema HW. Prevalence of Mycobacterium avium ssp. paratuberculosis infections in Canadian dairy herds. J Dairy Sci. 2018;101(12):11218-28. https://doi.org/10.3168/jds.2018-14854
Donat K, Kube J, Dressel J, Einax E, Pfeffer M, Failing K. Detection of Mycobacterium avium subspecies paratuberculosis in environmental samples by faecal culture and real-time PCR in relation to apparent within-herd prevalence as determined by individual faecal culture. Epidemiol Infect. 2015 Apr;143(5):975-85. https://doi.org/10.1017/S0950268814002465
Fleischer P, Kovarcik K, Slosarkova S. Paratuberkuloza – Pripravovany certifikacni program pro chovy dojeneho skotu [Paratuberculosis – An upcoming certification programme for dairy cattle herds]. Náš chov. 2018 Dec;78(12):55-8. Czech.
Garcia AB, Shalloo L. Invited review: The economic impact and control of paratuberculosis in cattle. J Dairy Sci. 2015 Aug;98(8):5019-39. https://doi.org/10.3168/jds.2014-9241
Kovarcik K, Kralova A. Paratuberkuloza – Aktualni nakazova situace v CR dle vysetreni bazenovych vzorku mleka [Paratuberculosis – The current epidemiological situation in the Czech Republic according to the examination of bulk tank milk samples]. Náš chov. 2018 Nov;78(11):33-5. Czech.
Kralik K, Slana I, Kralova A, Babak V, Whitlock RH, Pavlik I. Development of a predictive model for detection of Mycobacterium avium subsp. paratuberculosisin faeces by quntitative real time PCR. Vet Microbiol. 2011 Apr 21;149(1-2):133-8. https://doi.org/10.1016/j.vetmic.2010.10.009
Lombard JE, Wagner BA, Smith RL, McCluskey BJ, Harris BN, Payeur JB, Garry FB, Salman MD. Evaluation of environmental sampling and culture to determine Mycobacterium avium subspecies paratuberculosis distribution and herd infection status on US dairy operations. J Dairy Sci. 2006 Nov;89(11):4163-71. https://doi.org/10.3168/jds.S0022-0302(06)72461-4
OIE – The World Organisation for Animal Health. Manual of diagnostic tests and vaccines for terrestrial animals [Internet]. 2018 [accessed Aug 12, 2020). Available at: https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.01.15_PARATB.pdf.
Pillars RB, Grooms DL, Kaneene JB. Longitudinal study of the distribution of Mycobacterium avium subsp. paratuberculosis in the environment of dairy herds in the Michigan Johne’s disease control demonstration herd project. Can Vet J. 2009 Oct;50(10):1039-46.
Raizman EA, Wells SJ, Godden SM, Bey RF, Oakes MJ, Bentley DC, Olsen KE. The distribution of Mycobacterium avium ssp. paratuberculosis in the environment surrounding Minnesota dairy farms. J Dairy Sci. 2004 Sep;87(9):2959-66. https://doi.org/10.3168/jds.S0022-0302(04)73427-X
Rathnaiah G, Zinniel DK, Bannantine JP, Stabel JR, Grohn YT, Collins MT, Barletta RG. Pathogenesis, molecular genetics, and genomics of Mycobacterium avium subsp. paratuberculosis, the etiologic agent of Johne’s disease. Front Vet Sci. 2017 Nov 6;4: . https://doi.org/10.3389/fvets.2017.00187
Smith RL, Schuken YH, Pradhan AK, Smith JM, Whitlock RH, Van Kessel JS, Wolfgang DR, Grohn YT. Environmental contamination with Mycobacterium avium subsp. paratuberculosis in endemically infected dairy herds. Prev Vet Med. 2011 Oct 1;102(1): . https://doi.org/10.1016/j.prevetmed.2011.06.009
Schonenbrucher H, Abdulmawjood A, Failing K, Bulte M. New triplex real-time PCR assay for detection of Mycobacterium avium subsp. paratuberculosis in bovine faeces. Appl Environ Microbiol. 2008 May;74(9):2751-8. https://doi.org/10.1128/AEM.02534-07
Wolf R, Barkema HW, De Buck J, Orsel K. Sampling location, herd size, and season influence Mycobacterium avium ssp. paratuberculosis environmental cultures results. J Dairy Sci. 2015 Jan;98(1):275-87. https://doi.org/10.3168/jds.2014-8676