Immunization of cattle with synthetic peptides derived from the Boophilus microplus gut protein (Bm86).

Three synthetic peptides (SBm4912, SBm7462 and SBm19733), derived from the Bm86 glycoprotein from Boophilus microplus gut, were constructed and used to immunize cattle from a tick-free area. The immunized animals received three subcutaneous doses of the peptides, with saponin as adjuvant, at 30-day intervals. The immune response was evaluated by IgG elicited against the peptides by the detection of anti-Bm86 specific antibodies in situ and by Western blotting analysis. After tick challenge, reduction in the number, weight and oviposition capacity of engorged females was observed in the tick population that had fed on immunized animals. The results pointed a high efficacy (81.05%) for the SBm7462 synthetic peptide in relation to the others (p<0.01), demonstrating the efficiency of the immune response elicited by synthetic peptides to control the cattle tick B. microplus.

Exoantigens of an attenuated strain of Babesia bovis used as a vaccine against bovine babesiosis.

Bovine babesiosis caused by Babesia bovis remains a significant constraint to beef and milk cattle production throughout the world. Exoantigens released by the parasites in culture supernatants are a potential source of antigen to induce protective immunity. An attenuated strain of B. bovis from Brazil, catalogued as BbUFV1, was maintained in vitro by the MASP method, and exoantigen-containing supernatant fluids were collected daily to form a pool representing a 72-h culture cycle for preparation of the vaccine. Exoantigen concentration was estimated using a two-site EIA. Three groups of susceptible non-splenectomised male Bos taurus cattle, 14 months old, were used. Group A (vaccinated) received two subcutaneous immunizations with a 21-day interval of B. bovis supernatant, content 6500 EIA units of exoantigens plus 1.5 mg saponin, and Group B (adjuvant control) received two injections of adjuvant alone. Four weeks after the second immunization, Groups A, B and C (control) were challenged intravenously with 10(8) virulent parasites of a heterologous B. bovis strain. The results showed that exoantigens present in in vitro cultures can induce a high degree of protection against virulent heterologous challenge exposure. In Group A only one animal showed discrete parasitaemia; all developed a fever and slight decreases in PCV, with a rapid return to normal values. One animal of Group B died; the survivors showed fever, anaemia and parasitaemia. All animals of Group C died between 7 and 13 days after challenge. Vaccination elicited both humoral and cell-mediated immune responses. In Group A, after the challenge, the maximum antibody titer was 12,800. When vaccinated, cattle were tested at the moment of challenge for B. bovis-specific cell-mediated immunity by the monocytemigration inhibition test. A mean inhibition index of 60 +/- 0.33 was observed. Preliminary Western blot analysis of the immunogen revealed at least four proteins of molecular weight ranging between 30 and 160 kDa.

Antigenic profile of a pure isolate of Anaplasma marginale of Brazilian origin, using a western blot technique.

Anaplasma marginale initial bodies of the Brazilian isolate AUFV1 were purified from infected erythrocytes using a combination of lysis, ultrasonic disruption and differential centrifugation. Initial bodies were solubilised with a buffer containing protease inhibitors and non-ionic detergents. Immunochemical analysis by the Western blot technique revealed at least five proteins with apparent molecular weights (MW) of 105, 100, 97, 87 and 38 kDa when homologous sera were used as primary antibodies. Sera from cattle from Mato Grosso do Sul State in Brazil revealed five proteins of 105, 100, 87, 38 and 25 kDa; other heterologous sera obtained from Illinois, USA, bound to four antigens with MW of 105, 100, 87 and 38 kDa, the latter being stronger and broader than the others. No bands were observed in the non-infected erythrocyte control when the different A. marginale sera or antibodies against Babesia bovis and Babesia bigemina were used. Antibodies from cattle infected with the A. marginale isolates bound to proteins of 105, 100, 87 and 38 kDa, indicating that there are at least four peptides common to the isolates. The major surface proteins, designated MSP-1, MSP-2 and MSP-3, are also present in the Brazilian isolate AUFV1 and the sample from the National Research Centre of Beef Cattle in Mato Grosso do Sul State. One practical consideration of this study is the possibility of cross-protection between different Anaplasma isolates including some from Brazil.