Preparation and Evaluation of Alum Precipitate and Oil Adjuvant Multivalent Vaccines Against Clostridium perfringens

Abstract

Enterotoxaemia is one of the hazardous diseases of the livestock. In Pakistan prophylaxis failure is due to the vaccination with type D monovalent vaccine. There is a need to develop a cost effective multivalent vaccine against enterotoxaemia using characterized toxinotypes isolated from field. Indigenously (Punjab, Pakistan) characterized Clostridium perfringens toxinotypes A (MW551947.1), B (MW332247.1) and D (MW332258.1) (n=1 each) were used. These toxinotypes were used to produce higher amount of alpha, beta and epsilon toxin units under culture conditions. Colony forming units (CFU) of each bacterium was determined through the standard plate count method and 106 CFU/mL bacteria were used for vaccine dose. Monovalent, bivalent and multivalent oil adjuvant and alum precipitate vaccines were prepared. Formulated vaccines were passed the stability, sterility and safety test. Bacterin plus toxoid oil adjuvant vaccine produced higher (868.25±3.54 IU/mL) antibody titer at 28th day post vaccination in rabbits and 100% protection was observed after challenge. Multivalent bacterin plus toxoid oil adjuvant vaccine was used in field trials. Increased antibody response was detected after 4 months in sheep (1294.81±1.90 IU/mL) and goats (1091.85±2.51 IU/mL). During the experimental and field trials commercial vaccine did not produced higher antibody titer. Multivalent bacterin plus toxoid oil adjuvant vaccine proved as an excellent candidate for vaccination of animals against C. perfringens diseases, and it produced specific and efficient immune response to be used in field.

Keywords

• alpha
• bacterin
• beta
• epsilon
• toxoid
• vaccine