Experimental infection with Mycobacterium avium subspecies paratuberculosis resulting in decreased body weight in Holstein-Friesian calves.

Fifty calves inoculated at either 2 weeks or at 3, 6, 9, or 12 months of age with either a low or high dose of Mycobacterium avium subspecies paratuberculosis (MAP) were on average 32 and 39 kg lower in body weight, respectively, compared to negative controls at 17 months of age.

Chute du poids corporel chez des veaux Holstein-Friesian suite à l'infection expérimentale avecMycobacterium aviumsous-espèceparatuberculosis. Cinquante veaux qui ont été inoculés à l'âge de 2 semaines ou à l'âge de 3, 6, 9 ou 12 mois avec soit une dose faible ou élevée de Mycobacterium avium sous-espèce paratuberculosis (MAP) présentaient en moyenne un poids corporel inférieur de 32 kg et de 39 kg respectivement, comparativement aux témoins négatifs à l'âge de 17 mois.(Traduit par Isabelle Vallières).

Longitudinal evaluation of diagnostics in experimentally infected young calves during subclinical and clinical paratuberculosis.

Five calves were inoculated orally at 2 weeks of age with a dose of 5 × 10(9) colony-forming units of Mycobacterium avium subspecies paratuberculosis (MAP) on 2 consecutive days. Two calves developed clinical Johne's disease at 12 and 16 months of age after being consistently positive for MAP on fecal culture and antibody enzyme-linked immunosorbent assay (ELISA), starting 2 to 3 weeks and 4 to 5 months after inoculation, respectively.

Évaluation longitudinale du diagnostic lors de paratuberculose subclinique et clinique chez de jeunes veaux infectés expérimentalement. Cinq veaux ont été inoculés oralement à l'âge de 2 semaines avec une dose de 5 × 109 unités formatrices de colonies de Mycobacterium avium sous-espèce paratuberculosis (MAP) pendant 2 jours consécutifs. Deux veaux ont développé la maladie de Johne clinique à l'âge de 12 et de 16 mois après avoir obtenu des résultats constamment positifs pour MAP aux cultures de fèces et à l'ELISA, commençant entre 2 et 3 semaines et 4 et 5 mois après l'inoculation, respectivement.(Traduit par Isabelle Vallières).

Susceptibility to and diagnosis of Mycobacterium avium subspecies paratuberculosis infection in dairy calves: A review.

The primary objectives of paratuberculosis control programs are reducing exposure of calves to Mycobacterium avium subspecies paratuberculosis (MAP), reducing herd infection pressure and regular testing of cattle >36 months of age. Although control programs based on these principles have reduced prevalence of MAP infection in dairy herds, they have generally not eliminated the infection. Recent infection trial(s) have yielded new knowledge regarding diagnostic testing and age- and dose-dependent susceptibility to MAP infection. Calves up to 1 year of age are still susceptible to MAP infection; therefore, control programs should refrain from referring to specific ages with respect to susceptibility and prevention of new infections. Notwithstanding, lesions were more severe when calves were inoculated at 2 weeks versus 1 year of age. Furthermore, a high inoculation dose resulted in more pronounced lesions than a low inoculation dose, especially in young calves. Consequently, keeping infection pressure low should decrease the incidence of new MAP infections and severity of JD in cattle that do acquire the infection. It was also evident that early diagnosis of MAP infection was possible and could improve efficacy of control programs. Although its use will still need to be validated in the field, a combination of antibody ELISA and fecal culture in young stock, in addition to testing cattle >36 months of age when screening a herd for paratuberculosis, was expected to improve detection of dairy cattle infected with MAP. Although calves were inoculated using a standardized method in a controlled environment, there were substantial differences among calves with regards to immune response, shedding and pathology. Therefore, we inferred there were genetic differences in susceptibility. Important insights were derived from experimental infection trials. Therefore, it was expected that these could improve paratuberculosis control programs by reducing severity and incidence of JD by lowering infection pressure on-farm, and reducing exposure of young calves and older cattle. Furthermore, an earlier diagnosis could be achieved by combining ELISA and fecal shedding in young stock, in addition to testing cattle >36 months of age.

Shedding patterns of dairy calves experimentally infected with Mycobacterium avium subspecies paratuberculosis.

Although substantial fecal shedding is expected to start years after initial infection with Mycobacterium avium subspecies paratuberculosis (MAP), the potential for shedding by calves and therefore calf-to-calf transmission is underestimated in current Johne's disease (JD) control programs. Shedding patterns were determined in this study in experimentally infected calves. Fifty calves were challenged at 2 weeks or at 3, 6, 9 or 12 months of age (6 calves served as a control group). In each age group, 5 calves were inoculated with a low and 5 with a high dose of MAP. Fecal culture was performed monthly until necropsy at 17 months of age. Overall, 61% of inoculated calves, representing all age and dose groups, shed MAP in their feces at least once during the follow-up period. Although most calves shed sporadically, 4 calves in the 2-week and 3-month high dose groups shed at every sampling. In general, shedding peaked 2 months after inoculation. Calves inoculated at 2 weeks or 3 months with a high dose of MAP shed more frequently than those inoculated with a low dose. Calves shedding frequently had more culture-positive tissue locations and more severe gross and histological lesions at necropsy. In conclusion, calves inoculated up to 1 year of age shed MAP in their feces shortly after inoculation. Consequently, there is potential for MAP transfer between calves (especially if they are group housed) and therefore, JD control programs should consider young calves as a source of infection.

Dose-dependent interferon-gamma release in dairy calves experimentally infected with Mycobacterium avium subspecies paratuberculosis.

The interferon-gamma (IFN-γ) release assay is considered useful for diagnosis of subclinical paratuberculosis. However, interpretation can be subjective and complex; therefore, additional information regarding the course of the cellular immune response and effects of age and dose at infection would be helpful. Thirty-three calves were randomly allocated to 10 challenge groups and a negative control group. Calves were inoculated orally at 2 weeks or at 3, 6, 9, or 12 months of age. Within each age group, calves received either a high or low dose of Mycobacterium avium subspecies paratuberculosis (MAP). Monthly blood samples were collected, stimulated with Purified Protein Derivative (PPD) Johnin in vitro, and the subsequent release of IFN-γ measured. Calves inoculated with a high dose had earlier and stronger IFN-γ responses than low-dose calves. Furthermore, calves inoculated at 2 weeks of age produced less IFN-γ compared to those inoculated later in life. The IFN-γ response peaked (on average) 4 months after exposure; therefore, this would be an optimal interval to test cattle for MAP-infection (although the timing of field-based infections is unknown and clearance of infection a possibility). To conclude, the IFN-γ release assay could be a valuable diagnostic test on herd-level to indicate exposure to MAP.

Antibody response early after experimental infection with Mycobacterium avium subspecies paratuberculosis in dairy calves.

Serological testing in the early stages of Johne's disease has been successful using specific antigens and in-house ELISA. However, the use of a commercial ELISA has not been evaluated shortly after Mycobacterium avium subspecies paratuberculosis (MAP) infection, nor has it been determined whether this serological response is age or dose dependent. Fifty-six calves were randomly allocated to challenge groups (5 per group) and a negative control group. Calves were inoculated orally on 2 consecutive days at 2wk or at 3, 6, 9, or 12mo. Within each age group, 5 calves received either a high or low dose of MAP. Using a commercial ELISA, antibody responses were detected in 42% of the inoculated calves and were present in all age and dose groups (except for the 6-mo low-dose group). Antibody response profiles differed among individual calves; persistent as well as peak and bimodal peak responses existed. Calves inoculated at 12mo were ELISA positive within 4.5mo after inoculation, whereas those inoculated at younger ages took longer to become ELISA positive. Furthermore, calves inoculated with a high dose of MAP more often became ELISA positive than low-dose calves when inoculated at a younger age. In conclusion, a dose-dependent antibody response was detected by ELISA in a larger proportion of calves than expected soon after inoculation.

Evaluation of age-dependent susceptibility in calves infected with two doses of Mycobacterium avium subspecies paratuberculosis using pathology and tissue culture.

The longstanding assumption that calves of more than 6 months of age are more resistant to Mycobacterium avium subspecies paratuberculosis (MAP) infection has recently been challenged. In order to elucidate this, a challenge experiment was performed to evaluate age- and dose-dependent susceptibility to MAP infection in dairy calves. Fifty-six calves from MAP-negative dams were randomly allocated to 10 MAP challenge groups (5 animals per group) and a negative control group (6 calves). Calves were inoculated orally on 2 consecutive days at 5 ages: 2 weeks and 3, 6, 9 or 12 months. Within each age group 5 calves received either a high - or low - dose of 5 × 109 CFU or 5 × 107 CFU, respectively. All calves were euthanized at 17 months of age. Macroscopic and histological lesions were assessed and bacterial culture was done on numerous tissue samples. Within all 5 age groups, calves were successfully infected with either dose of MAP. Calves inoculated at < 6 months usually had more culture-positive tissue locations and higher histological lesion scores. Furthermore, those infected with a high dose had more severe scores for histologic and macroscopic lesions as well as more culture-positive tissue locations compared to calves infected with a low dose. In conclusion, calves to 1 year of age were susceptible to MAP infection and a high infection dose produced more severe lesions than a low dose.