Paratuberculosis (Johne's disease) has emerged as one of the most important diseases in cattle. The role of infected bull semen in the spread of infection remains unclear, as the correlation between the amount of excreted Mycobacterium avium subsp. paratuberculosis (semen and feces) and the infection load (blood and tissues) has not been defined. The aim of the present study was to study by culture, and a quantitative real-time polymerase chain reaction, the presence of bacteria in consecutive semen, blood, and fecal samples collected from one infected Piedmont breeding bull during a 380-day period. Five out of seven blood samples and all nine semen samples were positive in the real-time quantitative polymerase chain reaction with 101 to 102 and 102 to 104 copies of IS900/F57 per ml, respectively. In all, there were 9 fecal culture positive samples with too numerous to count colony forming units and positive real-time quantitative polymerase chain reactions ranging from 105 to 107 copies of IS900/F57. After the bull was euthanized, Mycobacterium avium subsp. paratuberculosis was cultured from various parts of the small and large intestines, liver tissue and lymph nodes and from the epididymis and vesicular glands. The results demonstrate a wide extraintestinal distribution of the bacterium and that breeding bulls should be considered a source of paratuberculosis infection due to their contact with other breeding bulls and a high number of heifers and cows through the natural mating process.

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of paratuberculosis (Johne's disease) in ruminants and is suggested to be one of the etiologic factors in Crohn's disease in humans. Contaminated milk might expose humans to that pathogen. The aim of the present study was to develop a novel real-time PCR assay providing the additional possibility to detect viable Mycobacterium avium subsp. paratuberculosis (MAP) based on the MAP-specific Mptb52.16 target. The design included an internal amplification control to identify false negative results.

Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease (JD) in ruminants and is shed into the milk of infected cows, which contributes to the controversial discussion about a possible link between MAP and Crohn's disease in humans. The aim of the study was to investigate the risk for the entry of MAP in the food chain via milk from dairy farms with subclinical JD. Therefore, the occurrence of MAP in the milk of a dairy herd with a low prevalence of JD was studied in single and bulk tank milk samples over a period of 23 mo and compared with MAP shedding into feces. Milk, fecal, and blood samples were taken from all cows older than 1.5 yr of age at the beginning and the end of the trial and analyzed for MAP or specific antibodies. In addition, 63 cows (33 MAP infected and 30 MAP noninfected) were selected for monthly sampling. Raw and pasteurized bulk tank milk samples were collected on a monthly basis. The milk samples were tested for MAP by real-time quantitative PCR (qPCR), and the fecal samples were tested for bacterial shedding by qPCR or solid culture. Based on the results of the herd investigations, the prevalence of cows shedding MAP was around 5%; no cases of clinical JD were observed during the study period. The results of the ELISA showed high variation, with 2.1 to 5.1% positive milk samples and 14.9 to 18.8% ELISA-positive blood samples. Monthly milk sampling revealed low levels of MAP shedding into the individual milk samples of both MAP-infected and noninfected cows, with only 13 cows shedding the bacterium into milk during the study period. Mycobacterium avium ssp. paratuberculosis was not detected by qPCR in any raw or pasteurized bulk tank milk sample throughout the study. A significant positive association could be found between MAP shedding into milk and feces. From the results of the present study, it can be concluded that MAP is only shed via milk in a small proportion of cows with subclinical JD for a limited period of time and is diluted below the detection level of qPCR within the bulk tank milk of these herds. These findings indicate that dairy herds subclinically infected with JD pose only a minor source for human MAP consumption with milk and milk products.

This study aimed to investigate the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) in small ruminants in Austria by testing 22,019 serum samples with ELISA for the presence of specific antibodies. Furthermore, detailed investigations in five MAP-infected goat herds were carried out by ELISA, qPCR and bacterial culture. The found animal-level apparent MAP seroprevalence was 2.0% for goats and 0.7% for sheep (calculated true prevalence 3.5% and 1.2%, respectively). Herd-level apparent MAP seroprevalence was 11.1% for goat herds and 8.9% for sheep flocks. Significant risk factors for seropositivity in goat herds were: herd size, animal trading, farmed as a dairy herd, Animal Health Service membership and cohabitation with farmed game. For sheep flocks, seroprevalence was significantly higher in flocks with animal trading and where cattle or goats were kept in the flock, respectively. The overall apparent within-herd MAP seroprevalence in the five goat farms investigated was 21.8% (11.7%–28.0%, calculated true seroprevalence 38.6%) and an overall rate of MAP shedding of 12.3% was detected (5.0%–24.7%). It was possible to identify MAP by culture using boot swab samples in each herd. The results indicated a moderate MAP infection rate in small ruminants in Austria.

Paratuberculosis (Johne's disease) in ruminants is caused by Mycobacterium avium subsp. paratuberculosis (MAP). Owing to the lack of accurate laboratory tests, diagnosis is challenging in subclinically infected cattle. To evaluate the long-term performance of serum ELISAs for the detection of paratuberculosis in a dairy herd with low MAP-prevalence, three investigations of all the cows and the consecutive testing of 33 cows suspected to be infected with MAP and 30 cows classified as MAP free were performed over a period of 22 months. Blood samples were tested by three commercial serum ELISAs, MAP shedding was detected by bacteriological culture and polymerase chain reaction (PCR). The ELISA results varied in a wide range in the herd investigations with 1.2% to 18.8% positive samples, the faecal samples were positive for MAP between 1.8% and 4.9% in the three herd investigations. Over the study period, ELISA-positive serum samples varied between 0.0% and 69.7% in MAP-suspicious and 0.0% and 17.6% in MAP-unsuspicious cows with a poor correlation between ELISAs and faecal shedding. The correlation coefficient of the optical density values of the three ELISAs varied between 0.348 and 0.61. Evidence of cow specific variations of residuals was found in all linear models. The linear mixed models showed relevant contribution of cow specific variation in explanation of the residual variances. They also showed significant effects of the explanatory ELISA, the group (MAP-suspicious or MAP-unsuspicious) and the time of sampling. It can be concluded that the choice of the laboratory test significantly influences the outcome of the testing for MAP and that none of the three ELISAs can be thoroughly recommended as single test for the early diagnosis of paratuberculosis in cattle. Test results should always be interpreted with caution to avoid erroneous decisions and the disappointment of those engaged in the abatement of paratuberculosis.