Antigenic analysis of Anaplasma marginale grown in bovine erythrocytes co-cultured with bovine endothelial cells.

Two monoclonal antibodies (mAbs) for A. marginale were used to test the antigenic integrity of A. marginale grown in vitro in bovine erythrocytes co-cultured with endothelial cells. Both the mAbs reacted in the indirect immunofluorescent antibody test with A. marginale grown in vitro and also detected the antigens in Western immunoblots of SDS-PAGE separated antigens made from A. marginale infected erythrocytes from the cultures. Furthermore, active replication was evident as [35S]-methionine is incorporated by A. marginale present in the second passage of a culture maintained for six weeks as shown by immunoprecipitation of labeled antigens by the mAbs. This indicates that A. marginale grown in the in vitro culture system described previously [Waghela et al., Vet. Parasitol. 73 (1997) 43] maintain antigenic character, and with further development the system can be used for preparing immunogens or diagnostic antigens.

A common high molecular weight antigen of Babesia bovis isolates from Mexico.

Cattle from an area of Mexico endemic with Babesia bovis infections have a dominant antibody response to a 152kDa antigen of the Tamaulipas strain of B. bovis. A mAb termed PB/5, showing a specific reactivity to this 152kDa antigen in Western blots, was identified. The mAb which reacted with the blunt end of B. bovis in an indirect fluorescent antibody test also reacted to a 152kDa antigen in two other isolates (Nuevo Leon and Yucatan), and a 175kDa antigen in the Huasteca B. bovis isolate from Mexico. Polyclonal monospecific sera from a calf inoculated with mAb-affinity purified 152kDa antigen (Tamaulipas strain) identified B. bovis by the indirect fluorescent antibody test and two antigens of B. bovis (65kDa and 152kDa) in Western blot. Since the epitope reacting to the mAb PB/5 is conserved, this antigen provides a basis for developing a diagnostic test or an immunogen.

Comparative studies of the antigenic polypeptide species VP4, VP6, and VP7 of three strains of bovine rotavirus.

Three bovine rotavirus strains belonging to two distinct serotype groups, serotype 6 (NCDV and B641) and B223, distinct from the other six mammalian rotavirus serotypes but not yet assigned to a serotype group, were compared with each other and with canine rotavirus (K9, serotype 3) by studying the properties of their cognate polypeptide species VP4, VP6, and VP7. The three viruses showed distinct differences in the polyacrylamide gel electrophoretic migration rates of protein species VP4 and VP7, with minor differences in VP6. Differences were also observed among the migration patterns of genome segments 4, 6, and the 7-8-9 triplet, which encode VP4, VP6, and VP7, respectively. Monoclonal antibodies (MAbs) to B223, which were directed against VP4 or VP7, showed homologous specificity for neutralization and immunofluorescence (IF), although one MAb reactive with VP4 also reacted by IF and by immunoprecipitation (IP) with all four viruses and weakly neutralized B641 and K9. This MAb may react with the epitope responsible for the B223-induced one-way neutralizing and protection response of calves against B641 observed in earlier studies. MAbs reactive with VP6 by IP showed enzyme-linked immunosorbent assay and IF reactivity with all three bovine viruses and the canine virus. The two serotype 6 viruses could be distinguished by the two B641 MAbs, B641-N2b reacting by neutralization and IF with both viruses and B641-N1 reacting with B641 and the serotype 3 canine rotavirus but not with NCDV. One nonneutralizing B641 MAb reacted by IP and IF with VP7 of all four rotaviruses examined, and one B223 MAb neutralized B223 and, to low titer, B641 and K9 although reacting by IP and IF with all four viruses. Three MAb-resistant mutants were selected by passage of B223 in the presence of one of three selected B223 MAbs at concentrations which only neutralized approximately 90% of the infectious virions. The resulting mutants were 100% resistant to neutralization with their respective MAb but remained neutralizable by the same selection of MAbs as the parent B223 virus.

Studies on rotavirus homologous and heterologous active immunity in infant mice.

Homologous and heterologous active immunity was studied in mice with mammalian group A rotaviruses. One day old mice were vaccinated with one of the following rotaviruses: bovine B641 (serotype 6), bovine B223 (untyped), simian SA11 (serotype 3) and murine EDIM (untyped). At 10 days of age they were challenged with EDIM virulent virus or SA11 virus. All the vaccines induced a serological antibody response in the mice but only the homologous immune response was protective.

Roles of predisposing factors and gonadal hormones in the buller syndrome of feedlot steers.

In a study of the role of predisposing factors and gonadal hormones in the buller syndrome, it was found that weather, amount of space available to each steer, and entry weights of the steers were not associated with increased occurrence of bulling; however, the type and timing of administration of the hormonal implant, the number of steers in a pen, and the manner in which the steers were grouped after arrival at the feedlot were found to influence the incidence of the syndrome. Serum concentrations of estradiol and testosterone in the buller steers were found to be lower during bulling than after a short period of isolation and apparent recovery from the syndrome. It was concluded that the occurrence of bulling is related to the use of hormonal implants and certain feedlot management procedures but that it is apparently not related to increased serum estrogen or testosterone concentrations.