A challenge experiment was performed to investigate whether administration of Mycobacterium avium subsp. paratuberculosis (MAP) via the respiratory route leads to MAP infection in calves. Eighteen calves from test negative dams were randomly allocated to four groups. Six calves were challenged with MAP nasally and six calves were challenged by transtracheal injection; three orally challenged calves served as positive controls, and three non challenged calves as negative controls. The challenge was performed as a nine-fold trickle dose, 107 CFU in total. Blood and faecal samples were collected frequently. Calves were euthanized three months post-challenge and extensively sampled. Blood samples were tested for the presence of antibodies and interferon gamma producing cells by ELISA. Faecal and tissue samples were cultured in a liquid culture system and the presence of MAP was confirmed by IS900 realtime PCR. Fourteen out of fifteen calves had no MAP antibody response. The negative controls remained negative; all positive controls became infected. Two nasally challenged calves showed a Purified Protein Derivative Avian (PPDA) specific interferon gamma response. In all nasally challenged calves, MAP positive intestinal samples were detected. In three calves of the nasal group MAP positive retropharyngeal lymph nodes or tonsils were detected. In all calves of the transtracheal group MAP positive intestinal tissues were detected as well and three had a MAP positive tracheobronchial lymph node. These findings indicate that inhalation of MAP aerosols can result in infection. These experimental results may be relevant for transmission under field conditions since viable MAP has been detected in dust on commercial dairy farms.

The influence of milk yield and milk composition on the diagnosis of Mycobacterium avium ssp. paratuberculosis (MAP) by milk ELISA in the context of the total IgG secretion patterns in milk throughout lactation and serum concentrations were investigated. A 2-yr trial was performed in which 1,410 dairy cows were sampled monthly and MAP milk ELISA status and milk yield and composition were determined. Data were analyzed by mixed model analysis. Milk yield was found to significantly influence ELISA results expressed as sample-to-positive (S/P) ratios. For each 5-kg increase in milk, the S/P ratio has to be multiplied by 0.89; therefore, high milk yield can change the MAP milk ELISA outcome of a cow in early infection from positive to negative. Parity influenced ELISA outcome significantly, indicating that cows with a parity >1 are more likely to be identified by milk testing. Also, herd was an important predictor, showing that herd prevalence influences the milk ELISA strongly. Other factors influencing the S/P ratios were protein concentration, somatic cell count, and days in milk. The IgG concentration and mass excreted per day were determined longitudinally in a subset of 41 cows of which samples and data of a complete lactation were available. Again, the IgG concentration in milk was mainly influenced by milk yield. The total IgG mass secreted per day in milk was found to be relatively constant, with a mean of 8.70 ± 5.38 g despite an increasing IgG concentration in serum at the same time. The variation of IgG concentration in milk can be mainly attributed to dilution through changes in milk yield. This supports the assumption that concentrations of MAP-specific antibodies are influenced by changes in milk yield similarly. In conclusion, we confirmed that antibody concentrations, and therefore MAP ELISA outcome, were influenced by milk yield, herd, and parity. To enhance performance, milk ELISA tests should either be performed in early or late lactation, when milk yield is low. From a management perspective, sampling should be done during early lactation before cows are bred again. Based on the slow progressive infection dynamics, only first-parity cows should be preferentially tested at the end of their first lactation to avoid false-negative results.

Mycobacterium avium subsp. paratuberculosis , the causative agent of Johne's disease in cattle, was identified in settled-dust samples of Dutch commercial dairy farms, both in the dairy barn and in the young stock housing. Bioaerosols may play a role in within-farm M. avium subsp. paratuberculosis transmission.

Vaccines against S. aureus bovine mastitis are scarce and show limited protection only. All currently available vaccines are applied via the parenteral (usually intramuscular) route. It is unknown, however, whether this route is the most suitable to specifically increase intramammary immunity to combat S. aureus at the site of infection. Hence, in the present study, immunization via mucosal (intranasal; IN), intramuscular (triangle of the neck; IM), intramammary (IMM) and subcutaneous (suspensory ligament; SC) routes were analyzed for their effects on the quantity of the antibody responses in serum and milk as well as the neutralizing capacity of the antibodies within serum. The experimental vaccine comprised the recombinant S. aureus immune evasion proteins extracellular fibrinogen-binding protein (Efb) and the leukotoxin subunit LukM in an oil-in-water adjuvant combined with a hydrogel and alginate. The highest titer increases for both Efb and LukM specific IgG1 and IgG2 antibody levels in serum and milk were observed following SC/SC immunizations. Furthermore, the harmful effects of Efb and leukotoxin LukMF’ on host-defense were neutralized by serum antibodies in a route-dependent manner. SC/SC immunization resulted in a significant increase in the neutralizing capacity of serum antibodies towards Efb and LukMF’, shown by increased phagocytosis of S. aureus and increased viability of bovine leukocytes. Therefore, a SC immunization route should be considered when aiming to optimize humoral immunity against S. aureus mastitis in cattle.

Mycobacterium avium subspecies paratuberculosis is the causative agent of paratuberculosis in cattle which causes a chronic infection of the small intestine. Since the transmission is only partly understood current control programs have been able to only decrease prevalence but not to eradicate disease from a herd. Unknown and therefore uncontrolled routes of transmission were suggested and infective bioaerosols were hypothesized as a potential candidate. This review gives an overview concerning disease transmission and focuses on consequences of bioaerosols on the within-herd transmission of paratuberculosis.