In Germany, the application of antimicrobial agents represents the method of choice to control infections of food-producing animals by bacteria of the genus Leptospira. However, only few data are available on the susceptibility of Leptospira spp. against antimicrobial agents and the in vivo efficacy of antimicrobial agents. Based on a modified broth microdilution method, a standardised in-vitro susceptibility testing system for Leptospira spp. was established which also included quality control aspects. Considering the minimum inhibitory concentrations determined in this study in-vivo efficacy is expected for the following classes of antimicrobial agents: penicillins, cephalosporins, macrolides, lincosamides, fluoroquinolones, phenicols, and in part also for aminoglycosides and tetracyclines. Clinical aspects and bioavailability of the drugs in the target tissues have to be taken into account. Penicillins and aminoglycosides as commonly used inhibitory supplements in semen extender seem to be effective against leptospires.