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The humoral immune response of dogs vaccinated with leptospiral pentavalent outer envelope and whole culture vaccines was monitored with the microscopic agglutination (MA) test and leptospiricidal activity (LA) test for a 2-year period. The leptospiral serovars in the vaccines was canicola, icterohaemorrhagiae, grippotyphosa, pomona, and hardjo. The LA test was markedly more sensitive than the MA test for detecting anti-Leptospira antibodies and was least noticeable with anti-pomona antibodies. Both Igm and IgG were produced in similar amounts in the vaccinated dogs, and both classes of immunoglobulins were reactive in the MA and LA tests. The IgM class of antibodies was slightly more reactive in the MA test, whereas IgG antibodies were somewhat more reactive in the LA test. A direct correlation between protective antibodies, as determined by the hamster passive-protection test, and antibodies reactive in the MA and LA tests was observed.

The hemolytic and sphingomyelinase C activities of supernatants of cultures of Leptospira interrogans serovar pomona tended to copurify when isoelectric fractionation was carried out. Both activities focused primarily at pH 8.1. Considered in conjunction with other circumstantial evidence, the results led to the conclusion that sphingomyelinase C is responsible for hemolysis.

The humoral immune response of cattle vaccinated with leptospiral pentavalent outer envelope and whole culture vaccines was monitored with the microscopic agglutination (MA) test and the leptospiricidal activity (LA) test for a 2.5-year period. The serovars present in the vaccines were canicola, icterohaemorrhagiae, grippotyphosa, pomona, and hardjo. The LA test had greater sensitivity than the MA test for detecting antibodies to the hardjo, canicola, and icterohaemorrhagiae components of the pentavalent vaccine and was approximately equal in sensitivity to the MA test for detecting antibodies to the grippotyphosa and pomona components of the leptospiral vaccine. Both IgM and IgG were produced in similar amounts in the immunized cattle, and both classes were reactive in the MA and the LA tests. The IgM class of antibodies was more reactive in the MA test, whereas the IgG class of antibodies was more reactive in the LA test. Using the MA test as the criterion of evaluating, the immunogenic potency of the individual vaccine components was found to be canicola more than icterohaemorrhagiae more than grippotyphosa more than pomona more than hardjo. A direct correlation between protective antibodies, as determined by the hamster passive protection test, and antibodies reactive in the MA and the LA tests was observed.

A protein-free medium composed of charcoal-detoxified Tweens (polysorbates), vitamins B12 and B1, inorganic salts, and organic buffer is described that supports the growth and subculture of pathogenic and saprophytic Leptospira. Growth was initiated from small inocula, and cell densities of 10(9) organisms per ml were attained. Antigenicity and immunogenicity of Leptospira cultivated in this medium were similar to those of cells cultivated in serum-containing media. The protein-free medium was converted to a low-protein medium by the addition of 0.1% bovine serum albumin.

Swine serologically negative for anti-Leptospira antibodies were given 2 doses of a pentavalent vaccine (3 weeks between doses) prepared from Leptospira serovars canicola, icterohaemorrhagiae, hardjo, pomona, and grip-potyphosa (0.2 mg/serovar/dose). Leptospires used for vaccinal production were cultivated in a protein-free medium or in a bovine albumin-containing medium. All vaccinated swine had demonstrable antibody titers within 1 week of the initial vaccination. Peak microscopic agglutination titers were between 256 and 1,024 after the 2nd vaccinal dose was given. After challenge exposure with serovar canicola, control swine had titers of at least 13,653 and the vaccinated swine had titers of 3,403 to 8,192, depending on the vaccine. Leptospiremia and kidney infections were not detected in any canicola Moulton immunized swine, but did appear in control swine. The Al(OH)3 adjuvant had no obvious influence of any of the vaccinal titers.