Freund's complete and incomplete adjuvants, preparation, and quality control standards for experimental laboratory animals use.

Quality control and quality assurance procedures are discussed for the agreed benchmark standard Freund's complete adjuvant (FCA). In addition, the use of the incomplete adjuvant (FIA) in the preparation of antisera is discussed. A major problem is the use of a safe and suitable mineral oil in FCA and FIA; manufacturers should provide infra-red spectra and gas liquid chromatography analyses. A range of safety tests, toxicity, pyrogenicity and endotoxin assays and advice on practical procedures for the use of these adjuvants are described.

Experimental immunisation and protection of guinea pigs with Vibrio cholerae toxoid and mucinases, neuraminidase and proteinase.

As measured by fluid accumulation in ileal loops, Vibrio cholerae mucinase complex, with or without toxoid, protected guinea pigs from challenges with V. cholerae live organisms and enterotoxin. The neuraminidase and proteinases of the complex were combined in modified oil emulsion or aluminum hydroxide adjuvants and the resultant vaccines given by the parenteral or oral routes. There was little difference between the two types of adjuvant. Control of stomach acidity improved oral vaccination. Animals injected intramuscularly (i.m.) with toxoid-containing vaccines were protected from challenge with cholera toxin (CT) whereas those given oral doses were not. Toxoid plus killed V. cholerae cells elicited a more effective protection against toxin challenge than killed V. cholerae cells alone. Vaccines containing mucinases, with or without toxoid, protected the animals from a live V. cholerae challenge. The anti-mucinase immune response may prevent adhesion of the V. cholerae cells and hence reduce delivery of toxin to receptors. These mucinases, neuraminidase and proteinases, may be useful components of acellular, toxoided cholera vaccines for human immunisation.