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In this study, products from all steps of anaerobic digestion at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle were examined and quantified for the presence of the causal agent of paratuberculosis, Mycobacterium avium subsp. paratuberculosis, using culture and quantitative real-time PCR (qPCR). Viable M. avium subsp. paratuberculosis cells were detected using culture in fermentors for up to 2 months; the presence of M. avium subsp. paratuberculosis DNA (10 1 cells/g) was demonstrated in all anaerobic fermentors and digestate 16 months after initiation of work at a biogas plant, using IS900 qPCR. F57 qPCR was able to detect M. avium subsp. paratuberculosis DNA (10 2 cells/g) at up to 12 months. According to these results, a fermentation process that extended beyond 2 months removed all viable M. avium subsp. paratuberculosis cells and therefore rendered its product M. avium subsp. paratuberculosis free. However, M. avium subsp. paratuberculosis DNA was found during all the examined periods (more than 1 year), which could be explained by either residual DNA being released from dead cells or by the presence of viable cells whose amount was under the limit of cultivability. As the latter hypothesis cannot be excluded, the safety of the final products of digestion used for fertilization or animal bedding cannot be defined, and further investigation is necessary to confirm or refute this risk.

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ABSTRACT:The objective of this study was to determine the wildlife hosts of Mycobacterium avium subsp. paratuberculosis (MAP) in the Czech Republic. A total of 8 796 wildlife animals were examined by culture of faecal or tissue samples during the years [2002][2003][2004][2005][2006][2007]. MAP was isolated from 12 (0.5%) out of 2 296 red deer (Cervus elaphus), two (0.2%) out of 835 roe deer (Capreolus capreolus), 78 (5.7%) out of 1 381 fallow deer (Dama dama), 28 (3.2%)out of 866 mouflons (Ovis musimon), four (2.5%) out of 162 chamois (Rupicapra rupicapra) and from one (0.1%) out of 805 wild boar (Sus scrofa). MAP was not cultured from 82 badgers (Meles meles), 55 martens (Martes foina), one pine marten (Martes martes), 25 brown hares (Lepus europaeus), five rabbits (Oryctolagus cuniculus), nine European polecats (Mustela putorius), two steppe polecats (Mustela eversmannii), two American minks (Mustela vison), four raccoon dogs (Nyctereutes procyonoides) and four Eurasian otters (Lutra lutra). MAP was isolated from three (2.0%) out of 149 small terrestrial mammals: one (5.9%) out of 17 brown rats (Rattus norvegicus), one (1.7%) out of 59 common voles (Microtus arvalis) and one (2.6%) out of 39 lesser white-toothed shrews (Crocidura suaveolens). Culture examinations of 34 house mice (Mus musculus) and 2 113 pigeons (Columba livia f. domestica) were negative. All 123 in vitro growing MAP isolates from wild ruminants were of IS900 RFLP type B-C1. One mouflon infected with a MAP strain which did not grow on the tested media was after IS1311-PRA-PCR assessed as being infected with a "sheep" strain. The RFLP type of the MAP isolate from the wild boar was of the RFLP type A-C10. Although the detection of MAP in wildlife in the Czech Republic was not very high, their role as a potential risk factor for cattle should be considered.

Background: The placenta is an important site for iron metabolism in humans. It transfers iron from the mother to the fetus. One of the major iron transport proteins is transferrin, which is a blood plasma protein crucial for iron uptake. Its localization and expression may be one of the markers to distinguish placental dysfunction.

ABSTRACTThe aim of this study was to monitor the persistence ofMycobacterium aviumsubsp.paratuberculosisin 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 forM. aviumsubsp.paratuberculosisby 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 meanM. aviumsubsp.paratuberculosiscell number of 3.09 × 103were positive forM. aviumsubsp.paratuberculosisbefore destocking compared to 43% with an average meanM. aviumsubsp.paratuberculosiscell number of 5.86 × 102after 24 months.M. aviumsubsp.paratuberculosis-positive samples were detected in the cattle barn as well as in the calf barn and surrounding areas.M. aviumsubsp.paratuberculosiswas detected from different matrices: floor and instrument scrapings, sediment, or scraping from watering troughs, waste pits, and cobwebs.M. aviumsubsp.paratuberculosisDNA 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 ofM. aviumsubsp.paratuberculosiscells were comparable.