The effect of DMSO and cultural behavior of two herpesviruses associated with Marek's disease.
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Biomedical subjects
Publications and source records attributed to R A Bankowski.
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Experiments were carried out to determine the usefulness of the egg-bit technique (EBT) in poultry virus research with various agents available on the market. The EBT was found satisfactory for propagating 9 strains of avian influenza virus of different antigenic groups, with only minor differences resulting when the procedure used two sources of gelatin, presence or absence of CaCl2, or commercial or specific-pathogen-free embryos.
Two serological tests, the virus-neutralization (VN) test in tissue culture using a tissue-cell-adapted virus and the enzyme-linked immunosorbent assay (ELISA), were compared to detect antibodies against Massachusetts 41 and Connecticut 46 strains of Infectious Bronchitis Virus (IBV). The VN test was conducted in wells of microplates by the usual procedure. The two strains of IBV were adapted after 20 serial passages to induce CPE in 24 hours in chickens embryos kidney cells (CEKC). The ELISA test was carried out using partially virus following ultracentrifugation of each stain of IBV as antigen. The ELISA test detected higher geometric mean antibody titers (GMT) against both strains of IBV than did the VN test. One hundred four serum samples taken at 1, 3, 5, 9, 22, 24, and 26 weeks of age from a flock of chickens vaccinated with the Mass strain three times and the Conn strain of IBV two times during the growing period showed higher antibody titer responses to the Conn 46 than to the Mass 41 strain. Maternal antibodies in chicks one week of age were readily detected by the ELISA test, whereas low or insignificant titers were found by the VN test. Sera of vaccinated chickens collected following challenge with Mass 41 or Conn 46 strain of IBV showed that the ELISA was more sensitive and showed higher titers than did the VN test. Although the VN test showed no rise in GMT in the same sera tested with the heterologous virus, the ELISA showed a slight increase or cross-reaction. The serum samples from the unchallenged control group showed no change in GMT with either test or IBV strain.
Experiments employing the various steps and reagents used in the enzyme-linked immunosorbent assay (ELISA) were conducted to produce an ELISA with the highest sensitivity and specificity possible for detecting Newcastle disease antibodies in chicken sera. Of the four types of antigen tested, crude antigen gave inconsistent results. However, an alcohol-precipitated antigen prepared from infectious allantoic-amniotic fluids was as satisfactory as more highly purified virus preparations. Other factors found to be extremely important were a 0.5M concentration of NaCl in the diluent and wash solutions used in the procedure, and a pH of 13 for sensitizing solution for maximum specific binding of the antigen to the microplate plastic wells. A comparison was made between the hemagglutination-inhibition (HI) titers of 550 known chicken sera and the corresponding ELISA values. Although the ELISA is much more sensitive than the HI test, there was a general but not a direct correlation between the two tests. The ELISA did not give more information than the HI test concerning protection against an NDV-induced drop in egg production. Preliminary observations indicated that this ELISA procedure is also applicable for turkey sera.
Parakeets were fed hulled millet seed containing 0.5% chlortetracycline (CTC) or minocycline. Blood concentrations of more than 1 micrograms CTC/ml and more than 5 micrograms minocycline/ml were obtained. Equivalent drug assay results were obtained from blood specimens collected by venipuncture or by use of treated filter-paper discs. The latter is a fairly simple method for assaying CTC concentrations in blood of treated psittacines.
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