Because of the fact that South Africa is a Newcastle disease virus (NDV)-endemic country, major concerns exist that the export of ostrich meat could transmit velogenic strains of this disease. The ability to transmit the virus could be reduced by effective vaccination of South African ostriches. In this study, two vaccination trials were conducted to assess serum antibody production in response to vaccination with La Sota strain NDV vaccines. To this end, a commercially available chicken anti-NDV enzyme-linked immunosorbent assay (ELISA) was modified for the detection of anti-NDV antibodies in ostrich serum. The results obtained with this ELISA were verified by comparison with an indirect ELISA. In the first trial, ostriches were immunized subcutaneously four times with different volumes of an inactivated vaccine and their immune response was determined from 2.5 mo up to the ideal slaughter age of 14 mo. Results indicated that ostriches responded in a dose-dependent manner and gave support for the vaccination schedule currently recommended to South African farmers. In a second trial, immunization by eyedrop with a live La Sota vaccine of 5-wk-old ostriches did not elicit a humoral immune response. The results indicate that it is highly unlikely that ostriches that have been vaccinated according to the recommended vaccination schedule can transmit the virus.