Three leptospiral bacterins, produced with different serovars of Serogroup Sejroe, namely the hardjo (bacterin A), wolffi (bacterin B) and guaricura (bacterin C), were evaluated in male hamsters (Mesocricetus auratus) by comparing the agglutinating and neutralizing antibodies titers using microscopic agglutination (MAT) and in vitro growth inhibition (GIT) tests. The immunization schedule was based on two 1.0 mL doses of non-diluted formalininactivated whole culture bacterin given through subcutaneous route with 10-day interval. The challenge was performed ten days after the second vaccine dose, when the animals were inoculated with 0.2 mL of non-inactivated cultures of each serovar through intraperitoneal route. On the 21st post-challenge day (PCD), all animals were bled and their sera were joined in pools (n=8) and tested by MAT and GIT. All vaccinated and control animals presented no clinical signs of leptospirosis after the challenge, but the serovar guaricura was isolated from the kidneys of control animals on the 21st PCD. The MAT results showed cross agglutinins between serovars hardjo and wolffi, and between wolffi and guaricura. The GIT results revealed the presence of cross neutralizing antibodies between serovars wolffi or guaricura against hardjo, wolffi and guaricura. It was found that the tested strain of serovar hardjo did not produce detectable levels of neutralizing antibodies, indicating its poor immunogenicity.