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.

Veterinary Research Institute Brno Czech Republic

Zobrazit více v PubMed

Aly SS, et al. 2010. Correlation between Herrold egg yolk medium culture and real-time quantitative polymerase chain reaction results for Mycobacterium avium subspecies paratuberculosis in pooled fecal and environmental samples. J. Vet. Diagn. Invest. 22:677–683 PubMed

Berghaus RD, Farver TB, Anderson RJ, Jaravata CC, Gardner IA. 2006. Environmental sampling for detection of Mycobacterium avium ssp. paratuberculosis on large California dairies. J. Dairy Sci. 89:963–970 PubMed

Cook KL, Britt JS. 2007. Optimization of methods for detecting Mycobacterium avium subsp. paratuberculosis in environmental samples using quantitative, real-time PCR. J. Microbiol. Methods 69:154–160 PubMed

Cossu A, et al. 2011. MAP3738c and MptD are specific tags of Mycobacterium avium subsp. paratuberculosis infection in type I diabetes mellitus. Clin. Immunol. 141:49–57 PubMed

Cossu D, et al. 2011. Association of Mycobacterium avium subsp. paratuberculosis with multiple sclerosis in Sardinian patients. PLoS One 6:e18482 doi:10.1371/journal.pone.0018482 PubMed DOI PMC

Eamens GJ, et al. 2007. Evaluation of radiometric faecal culture and direct PCR on pooled faeces for detection of Mycobacterium avium subsp. paratuberculosis in cattle. Vet. Microbiol. 125:22–35 PubMed

Eisenberg SWF, et al. 2010. Detection of spatial and temporal spread of Mycobacterium avium subsp. paratuberculosis in the environment of a cattle farm through bio-aerosols. Vet. Microbiol. 143:284–292 PubMed

England LS, Lee H, Trevors JT. 1997. Persistence of Pseudomonas aureofaciens strains and DNA in soil. Soil Biol. Biochem. 29:1521–1527

Fritsch I, Luyven G, Kohler H, Lutz W, Mobius P. 2012. Suspicion of Mycobacterium avium subsp. paratuberculosis transmission between cattle and wild-living red deer (Cervus elaphus) by multitarget genotyping. Appl. Environ. Microbiol. 78:1132–1139 PubMed PMC

Grewal SK, Rajeev S, Sreevatsan S, Michel FC., Jr 2006. Persistence of Mycobacterium avium subsp. paratuberculosis and other zoonotic pathogens during simulated composting, manure packing, and liquid storage of dairy manure. Appl. Environ. Microbiol. 72:565–574 PubMed PMC

Gumber S, Taylor DL, Marsh IB, Whittington RJ. 2009. Growth pattern and partial proteome of Mycobacterium avium subsp. paratuberculosis during the stress response to hypoxia and nutrient starvation. Vet. Microbiol. 133:344–357 PubMed

Hasonova L, Pavlik I. 2006. Economic impact of paratuberculosis in dairy cattle herds: a review. Vet. Med. 51:193–211

Jorgensen JB. 1977. Survival of Mycobacterium paratuberculosis in slurry. Nord. Vet. Med. 29:267–270 PubMed

Kaevska M, et al. 2011. “Mycobacterium avium subsp. hominissuis” in neck lymph nodes of children and their environment examined by culture and triplex quantitative real-time PCR. J. Clin. Microbiol. 49:167–172 PubMed PMC

Kawaji S, Begg DJ, Plain KM, Whittington RJ. 2011. A longitudinal study to evaluate the diagnostic potential of a direct faecal quantitative PCR test for Johne's disease in sheep. Vet. Microbiol. 148:35–44 PubMed

Kopecna M, et al. 2008. Distribution and transmission of Mycobacterium avium subspecies paratuberculosis in farmed red deer (Cervus elaphus) studied by faecal culture, serology and IS900 RFLP examinations. Vet. Med. 53:510–523

Kopecna M, et al. 2008. The wildlife hosts of Mycobacterium avium subsp. paratuberculosis in the Czech Republic during the years 2002–2007. Vet. Med. (Praha) 53:420–426

Kralik P, et al. 2011. Development of a predictive model for detection of Mycobacterium avium subsp. paratuberculosis in faeces by quantitative real time PCR. Vet. Microbiol. 149:133–138 PubMed

Kudykina YK, Shleeva MO, Artsabanov VY, Suzina NE, Kaprelyants AS. 2011. Generation of dormant forms by Mycobacterium smegmatis in the poststationary phase during gradual acidification of the medium. Microbiology 80:638–649 PubMed

Lamont EA, Bannantine JP, Armien A, Ariyakumar DS, Sreevatsan S. 2012. Identification and characterization of a spore-like morphotype in chronically starved Mycobacterium avium subsp. paratuberculosis cultures. PLoS One 7:e30648 doi:10.1371/journal.pone.0030648 PubMed DOI PMC

Lu Z, et al. 2008. The importance of culling in Johne's disease control. J. Theor. Biol. 254:135–146 PubMed

Man SM, Kaakoush NO, Mitchell HM. 2011. The role of bacteria and pattern-recognition receptors in Crohn's disease. Nat. Rev. Gastroenterol. Hepatol. 8:152–168 PubMed

Marsh P, Morris NZ, Wellington EMH. 1998. Quantitative molecular detection of Salmonella typhimurium in soil and demonstration of persistence of an active but non-culturable population. FEMS Microbiol. Ecol. 27:351–363

McKenna SL, Keefe GP, Tiwari A, VanLeeuwen J, Barkema HW. 2006. Johne's disease in Canada part II: disease impacts, risk factors, and control programs for dairy producers. Can. Vet. J. 47:1089–1099 PubMed PMC

Moravkova M, et al. 2008. Strategy for the detection and differentiation of Mycobacterium avium species in isolates and heavily infected tissues. Res. Vet. Sci. 85:257–264 PubMed

Mura M, et al. 2006. Replication and long-term persistence of bovine and human strains of Mycobacterium avium subsp. paratuberculosis within Acanthamoeba polyphaga. Appl. Environ. Microbiol. 72:854–859 PubMed PMC

Ogram AV, Mathot ML, Harsh JB, Boyle J, Pettigrew CA. 1994. Effects of DNA polymer length on its adsorption to soils. Appl. Environ. Microbiol. 60:393–396 PubMed PMC

Pickup RW, et al. 2006. Mycobacterium avium subsp. paratuberculosis in lake catchments, in river water abstracted for domestic use, and in effluent from domestic sewage treatment works: diverse opportunities for environmental cycling and human exposure. Appl. Environ. Microbiol. 72:4067–4077 PubMed PMC

Pillars RB, Grooms DL, Kaneene JB. 2009. 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. 50:1039–1046 PubMed PMC

Pribylova R, et al. 2011. Soil and plant contamination with Mycobacterium avium subsp. paratuberculosis after exposure to naturally contaminated mouflon feces. Curr. Microbiol. 62:1405–1410 PubMed

Raizman E. A., et al. 2011. Leaching of Mycobacterium avium subsp. paratuberculosis in soil under in vitro conditions. Vet. Med. Int. 2011:506239 doi:10.4061/2011/506239 PubMed DOI PMC

Raizman EA, et al. 2004. The distribution of Mycobacterium avium ssp. paratuberculosis in the environment surrounding Minnesota dairy farms. J. Dairy Sci. 87:2959–2966 PubMed

Romanowski G, Lorenz MG, Wackernagel W. 1991. Adsorption of plasmid DNA to mineral surfaces and protection against DNase I. Appl. Environ. Microbiol. 57:1057–1061 PubMed PMC

Romanowski G, Lorenz MG, Wackernagel W. 1993. Plasmid DNA in a groundwater aquifer microcosm-adsorption, DNase resistance and natural genetic-transformation of Bacillus subtilis. Mol. Ecol. 2:171–181 PubMed

Romanowski G, Lorenz MG, Wackernagel W. 1993. Use of polymerase chain-reaction and electroporation of Escherichia coli to monitor the persistence of extracellular plasmid DNA introduced into natural soils. Appl. Environ. Microbiol. 59:3438–3446 PubMed PMC

Salgado M, et al. 2011. Fate of Mycobacterium avium subsp. paratuberculosis after application of contaminated dairy cattle manure to agricultural soils. Appl. Environ. Microbiol. 77:2122–2129 PubMed PMC

Shleeva MO, et al. 2002. Formation and resuscitation of “non-culturable” cells of Rhodococcus rhodochrous and Mycobacterium tuberculosis in prolonged stationary phase. Microbiology 148:1581–1591 PubMed

Singh AV, Singh SV, Singh PK, Sohal JS, Singh MK. 2011. High prevalence of Mycobacterium avium subspecies paratuberculosis (“Indian bison type”) in animal attendants suffering from gastrointestinal complaints who work with goat herds endemic for Johne's disease in India. Int. J. Infect. Dis. 15:e677–e683 PubMed

Slana I, Kralik P, Kralova A, Pavlik I. 2008. On-farm spread of Mycobacterium avium subsp. paratuberculosis in raw milk studied by IS900 and F57 competitive real time quantitative PCR and culture examination. Int. J. Food Microbiol. 128:250–257 PubMed

Slana I, Pribylova R, Kralova A, Pavlik I. 2011. Persistence of Mycobacterium avium subsp. paratuberculosis at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle. Appl. Environ. Microbiol. 77:3115–3119 PubMed PMC

Smith RL, et al. 2011. Environmental contamination with Mycobacterium avium subsp. paratuberculosis in endemically infected dairy herds. Prev. Vet. Med. 102:1–9 PubMed

Tuci A, et al. 2011. Fecal Detection of Mycobacterium avium paratuberculosis using the IS900 DNA sequence in Crohn's disease and ulcerative colitis patients and healthy subjects. Dig. Dis. Sci. 56:2957–2962 PubMed

van Roermund HJW, Bakker D, Willernsen PTJ, de Jong MCM. 2007. Horizontal transmission of Mycobacterium avium subsp. paratuberculosis in cattle in an experimental setting: calves can transmit the infection to other calves. Vet. Microbiol. 122:270–279 PubMed

Whittington RJ, Marsh IB, Reddacliff LA. 2005. Survival of Mycobacterium avium subsp. paratuberculosis in dam water and sediment. Appl. Environ. Microbiol. 71:5304–5308 PubMed PMC

Whittington RJ, et al. 2003. Isolation of Mycobacterium avium subsp. paratuberculosis from environmental samples collected from farms before and after destocking sheep with paratuberculosis. Aust. Vet. J. 81:559–563 PubMed

Whittington RJ, Marshall DJ, Nicholls PJ, Marsh AB, Reddacliff LA. 2004. Survival and dormancy of Mycobacterium avium subsp. paratuberculosis in the environment. Appl. Environ. Microbiol. 70:2989–3004 PubMed PMC

Wu CW, et al. 2009. A novel cell wall lipopeptide is important for biofilm formation and pathogenicity of Mycobacterium avium subspecies paratuberculosis. Microb. Pathog. 46:222–230 PubMed PMC

Young JS, Gormley E, Wellington EMH. 2005. Molecular detection of Mycobacterium bovis and Mycobacterium bovis BCG (Pasteur) in soil. Appl. Environ. Microbiol. 71:1946–1952 PubMed PMC

Nontuberculous Mycobacteria Prevalence in Aerosol and Spiders' Webs in Karst Caves: Low Risk for Speleotherapy

An intra-laboratory cultural and real-time PCR method comparison and evaluation for the detection of subclinical paratuberculosis in dairy herds

Spread of Mycobacterium avium subsp. paratuberculosis through soil and grass on a mouflon (Ovis aries) pasture