Browsing by Author "De Wet, Bertus"

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    Evaluation of DNA vaccines developed against Mycoplasma struthionis sp. nov. str. Ms01 in ostriches
    (Stellenbosch : Stellenbosch University, 2016-03) De Wet, Bertus; Botes, Annelise; Bellstedt, D. U.; Stellenbosch University. Faculty of Science. Dept. of Biochemistry.
    ENGLISH ABSTRACT: The vast demand for ostrich meat has made South Africa the leader in not only the production of ostrich meat but also ostrich associated products such as feathers and leather. Ostrich specific mycoplasmas (referred to as Ms01, Ms02 and Ms03) cause respiratory tract infections with subsequent reduction in physical growth rate and therefore reduced production. To date no vaccines are available to combat these infections in ostriches. In this laboratory three DNA vaccines (pCI-neo, VR1012 and VR1020) have been developed with each containing the Ms01 oppA gene as antigen. The aim of this study was to evaluate these developed DNA vaccines in a mammalian cell culture based system as well as an ostrich vaccination trial. COS-1 cells were transfected with the three developed DNA vaccines. Transcription of the oppA gene was proven for all the plasmids. Translation into the OppA protein was shown to be limited to the VR1020_oppA plasmid. The protein was visualised by SDS-PAGE and detected by western blot using chemiluminescence. Two of the vaccines, VR1020_oppA and pCI-neo_oppA, were administered in three concentrations (100 μg/ml, 300 μg/ml and 600 μg/ml) to ostriches during a vaccination trial followed by a booster injection. The ability of the vaccines to elicit anti-OppA antibodies was measured using ELISA. The pCI-neo_oppA vaccine failed to induce an immune response against the antigen after both the first and booster vaccinations. The VR1020_oppA vaccine on the other hand was able to elicit an anti-OppA immune response.