DP020 Establishment of a Capture-ELISA to detect the Beta2-toxin of Clostridium perfringens

Abstract

A selected water-in-oil (W/O) emulsion was evaluated in terms of stability and antibody-response and side-effects in experimental animals.The results from stability tests performed at 4°C and 37°C indicate a storability of the W/O emulsion of more than a year under normal storage conditions. The antibody-response of rabbits after injection of E.coli K99 antigen and bovine serum albumin (BSA) using the W/O emulsion as vehicle was generally significantly higher than the response to the same antigens injected within incomplete Freund's adjuvant (IFA): twenty days after the first antigen injection the rabbits injected with antigen in the W/O emulsion showed average anti-K99 and anti-BSA titres which were ca 5x as high as the titres in sera from rabbits injected with the antigens in IFA. Thirty-two and forty-two days after the first antigen injections the W/O group showed anti-K99 titres which were respectively ca 3.5x and 2x higher than titres in the IFA group, while anti-BSA titres in the W/O group were respectively ca 13x and 6.5x higher than in the IFA group.Intra-muscular and/or subcutaneous injection of the W/O emulsion in rabbits and pigs resulted in a slight degeneration of local tissue. When antigens were present in the emulsion granulomatas were often observed. The rise in temperature in pigs after injection of killed E.coli 0149:K91,K88 in the W/O emulsion was comparable with that induced by E.coli in physiological saline.From these results it is concluded that the W/O emulsion is likely to be a promising basis for veterinary vaccines and that the application of the W/O emulsion may contribute to the limitation of the number of animals used for raising antibodies for general immunological purposes.