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Abstract. The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis-positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.

We explored the relationship between the level of bovine leukemia virus antibodies and provirus load during natural infection. For that purpose, a set of 50 blood and milk paired samples were analyzed for the presence of bovine leukemia virus provirus and antibodies. Additionally, provirus load and antibody titers were measured and the relationship between these variables was investigated. Bovine leukemia provirus was detected in 59% of milk samples and a negative correlation was observed between the level of milk provirus load and milk antibody titers. By the consumption of raw milk, calves might be exposed to bovine leukemia virus favoring the perinatal transmission of this disease.

The effect of monensin on milk production was evaluated in 58 lactating Holstein cows (48 multiparous; 10 primiparous) grazing a mixed-alfalfa pasture and supplemented with a partial mixed ration in a completely randomized design with repeated measurements. Cows were paired by calving date, lactation number, previous lactation milk production, body weight, and body condition score and were assigned to one of 2 treatments: control or monensin. Cows on the monensin treatment received 2 monensin controlled-release capsules (335 mg/d for 90 d), one 30 d before the expecting calving date and the other 60 d after calving. Short-term (0 to 150 d in milk) and long-term (305-d adjusted lactation) effects of monensin were evaluated. Pasture (measured by difference between pre- and postgrazing pasture mass), supplements, and total dry matter intake did not differ between treatments and averaged 8.7, 14.1, and 22.9 kg/d, respectively. In the short-term, monensin increased milk production (27.7 vs. 26.6 kg/d) and milk protein yield (0.890 vs. 0.860 kg/d); milk fat yield was not affected (0.959 kg/d). Monensin decreased milk fat content (3.51 vs. 3.60%) with no changes in milk protein content (3.25%). In the long term, milk production and milk protein yield were also increased by monensin: 214 and 7 kg, respectively. Monensin reduced the loss of body condition score and increased percentage of pregnancy at first service (44.8 vs. 20.7%). Monensin improves production and reproduction performance of dairy cows grazing a mixed-alfalfa pasture and supplemented with a partial mixed ration.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a major cause of intestinal disease and hemolytic uremic syndrome, a serious systemic complication that particularly affects children. Cattle are primary reservoirs for EHEC O157:H7 and the main source of infection for humans. Vaccination of cattle with different combinations of bacterial virulence factors has shown efficacy in decreasing EHEC O157:H7 shedding. It is, therefore, important to demonstrate whether vaccination of pregnant cows with EHEC O157:H7 induces high titers of transferable antibodies to avoid early colonization of calves by the bacteria. In this study we evaluated the ability of EspA, EspB, the C-terminal fragment of 280 amino acids of γ-intimin (γ-intimin C₂₈₀) and inactivated Shiga toxin (Stx) 2 proteins to induce specific antibodies in colostrum and their passive transference to colostrum-fed calves. Friesian pregnant cows immunized by the intramuscular route mounted significantly high serum and colostrum IgG responses against EspB and γ-intimin C₂₈₀ that were efficiently transferred to their calves. Antibodies to EspB and γ-intimin C₂₈₀ were detected in milk samples of vaccinated cows at d 40 postparturition. Significant Stx2-neutralizing titers were also observed in colostrum from Stx2-vaccinated cows and sera from colostrum-fed calves. The results presented showed that bovine colostrum with increased levels of antibodies against EHEC O157:H7 may be obtained by systemic immunization of pregnant cows, and that these specific antibodies are efficiently transferred to newborn calves by feeding colostrum. Hyperimmune colostrum and milk may be an alternative to protect calves from early colonization by EHEC O157:H7 and a possible key source of antibodies to block colonization and toxic activity of this bacterium.