A pair of synthetic oligonucleotide primers, designed from the gene encoding a 32-kDa intraerythrocytic piroplasm surface protein of Theileria sergenti, were used to amplify parasite DNA from the blood of T. sergenti-infected cattle by means of the polymerase chain reaction (PCR). PCR-amplified DNA was examined by electrophoresis and by dot blot or microplate hybridization using a parasite-specific cDNA probe. PCR was specific for T. sergenti, since no amplification was detected with DNA from Anaplasma centrale, Babesia ovata, uninfected erythrocytes, and leukocytes. This method was sensitive enough to detect about 4.5 parasites per I.al of blood with a 10-p.l sample volume. Moreover, of 66 specimens from grazing cattle, 40 were microscopically positive, whereas PCR revealed that 54 samples were positive. Therefore, PCR provides a useful diagnostic tool for detecting T. sergenti-infected cattle, and it is significantly more sensitive than the current methods.
Three pregnant cows were inoculated intramuscularly with inactivated vaccine to bovine rotavirus (BRV) serotype 1 (BRV-1) and serotype 2 (BRV-2). Serum neutralizing antibody (NA) titers against both serotypes increased significantly after immunization. NA titers of colostrum obtained from immunized cows against BRV-1 and BRV-2 were 29286 and 38109, respectively, which were significantly higher than those from non-immunized control cows. Nine and 6 colostrum deprived calves were orally challenged with BRV-1 and BRV-2, respectively, and monitored for clinical manifestation and viral shedding. Five calves of them, 3 with BRV-1 and 2 with BRV-2, received 2 l of milk replacer supplemented with 10% immune colostrum 2 hr before challenge and twice daily for the first 5 days after challenge. Other 10 calves, 6 with BRV-1 and 4 with BRV-2, were fed only milk replacer as controls. All control calves developed severe diarrhea and shed a large amount of BRV in feces, beginning from 24 to 48 hr after challenge inoculation. On the contrary, all calves but one fed colostrum supplement remained clinically healthy after challenge, and BRV was not detected in their feces during feeding immune colostrum. The possibility that continuous feeding of immune colostrum is capable of preventing newborn calves from diarrhea associated with BRV and viral shedding was suggested.
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