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Isolation from Pasteurella multocida of a Lipopolysaccharide Antigen with Immunizing and Toxic Properties

National Animal Disease Laboratory, Animal Disease and Parasite Research Division, Agricultural Research Service, Ames, Iowa

ABSTRACT

A heat-stable, particulate, lipopolysaccharide-protein antigenic complex has been isolated from a virulent, encapsulated strain of Pasteurella multocida by extraction with cold, formalinized saline, and centrifugation at 105,000 x g. The original bacterial culture had been obtained from a bison that died of hemorrhagic septicemia, an infectious disease of cattle and buffalo. Injection of fractional milligram amounts of the antigen into mice, rabbits, and calves produced toxic reactions which frequently resulted in death of the host. The surviving animals demonstrated a high degree of immunity to challenge with live, virulent organisms. Two injections with 15 µg of the antigen produced a high degree of immunity in mice without the development of any signs of toxicity. The gross chemical composition and toxicity of the antigen were similar to those reported for endotoxins obtained by the Boivin or Westphal procedure. Although strong serological cross-reactions were obtained in Ouchterlony plates between the 105,000 x g antigens from the bison strain and an avian strain with antisera to these strains, these antisera agglutinated only the bacterial cells of the homologous strain. The active immunity obtained in mice by the injection of the 105,000 x g antigens of each strain was specific and could be correlated with the agglutination tests.