Three antigens were prepared from a type-3 avian strain of Pasteurella multocida, and their chemical and immunologic characteristics were studied. An antigen, designated 2.5S, was extracted with 2.5% NaCl solution and purified by chromatography. Lipopolysaccharide (LPS) was extracted with phenol-water, and a third antigen, designated FS, was extracted in 0.3% formalin solution containing 0.85% NaCl and purified by differential centrifugation. The 2.5S and the FS antigens consisted of 40% protein and 15% carbohydrate, whereas LPS did not contain a substantial amount of protein. A major protein component with a molecular weight of 44,000 was detected in the 2.5S antigen, as well as in the FS antigen. Of the 3 antigens, LPS had thehighest activity in mouse lethality and Limulus lysate tests. Antigenic cross-reactions among the 3 antigens were demonstrated by immunodiffusion tests. The 2.5S antigen was indistinguishable from the FS antigen, as both antigens contained the LPS component of approximately 45%. Treatments with various reagents indicated that the 2.5S and FS antigens contained at least 2 antigenic determinants. The first was a heat-stable protein sensitive to protease or phenol-water, and the second was a periodate-sensitive carbohydrate, which was an major antigenic determinant on the LPS antigen.

A strain of Pasteurella multocida of avian origin expressed high molecular weight outer membrane proteins when grown in turkey plasma or in brain-heart infusion broth containing the iron chelator dipyridyl. The proteins were not detected when this strain was grown in brain-heart infusion broth or in brain-heart broth containing dipyridyl and excess iron.

Three antigens were prepared from Pasteurella multocida strain P-1059, and their immunogenicity and antigenic relationships were investigated. The 3 antigens were a soluble antigen purified from a 2.5% NaCl extract (2.5S), a similar antigen purified from an extract in 0.3% formalin solution containing 0.85% NaCl (FS), and lipopolysaccharide (LPS). The antigens were treated with various chemicals and enzymes to study their antigenic and immunogenic determinants. Antigenic analyses with ELISA inhibition tests indicated that 2.5S and FS were similar LPS-protein complex antigens. The 2.5S and FS antigens induced protective immunity in turkeys with high antibody titers against LPS antigen. Although LPS was a component of 2.5S and FS,LPS itself was poorly immunogenic in turkeys. The antigenicity of protein compounds in 2.5S was deteriorated by protease treatment, which, however, did not significantly diminish the protective immunogenicity. Treatment of 2.5S with sodium periodate, altering its carbohydrate moieties, decreased its immunogenicity. The immunogenicity of 2.5S also was abolished by phenol-water treatment, owing to dissociation of the LPS-protein complex. These findings suggest that a certain form of LPS-protein complex is essential for the induction of immunity against the P multocida infection in turkeys.

The respiratory tract of healthy chickens contain few free-residing phagocytic cells. Intratracheal inoculation with Pasteurella multocida stimulated a significant (P < 0.05) migration of cells to the lungs and air sacs of White Rock chickens within 2 hours after inoculation. We found the maximal number of avian respiratory tract phagocytes (22.9 +/- 14.0 x 10(6)) at 8 hours after inoculation. Flow cytometric analysis of these cells revealed 2 populations on the basis of cell-size and cellular granularity. One of these was similar in size and granularity to those of blood heterophils. Only this population was capable of generating oxidative metabolites in response to phorbol myristate acetate. The ability of the heterophils to produce hydrogen peroxide, measured as the oxidation of intracellularly loaded 2', 7'-dichlorofluorescein, decreased with time after inoculation. These results suggest that the migration of heterophils, which are capable of high levels of oxidative metabolism, to the lungs and air sacs may be an important defense mechanism of poultry against bacterial infections of the respiratory tract.