Mice infected in the left hind footpad with 5 log 10 acid-fast bacilli of Mycobacterium ulcerans were divided into an untreated control group and 17 treatment groups that received one of the following regimens for 4 weeks (all doses in milligrams per kilogram): 100 mg of azithromycin (AZM), 100 mg of clarithromycin (CLR), or 50 mg of AZM for a duration of 5 days a week (daily), three times a week, or once weekly. In addition, the following regimens were administered daily: 100 mg of telithromycin (TLM), sparfloxacin (SPX), or moxifloxacin (MOX); 200 mg of levofloxacin (LVX); 100 mg of streptomycin (STR) or amikacin (AMK); 10 mg of rifampin (RIF); and the combination of 10 mg of RIF and 100 mg of AMK (RIF؉AMK). After completion of treatment, mice were observed for 30 weeks. The effectiveness of treatment regimens was assessed in terms of the delay in median time to footpad swelling in treated mice compared with that in the untreated controls. Clear-cut bactericidal activity, i.e., an observed delay in footpad swelling that exceeded the period of treatment, was observed in the STR-, AMK-, and RIF؉AMK-treated mice. However, all mice treated with either AMK or STR alone had swollen footpads before the end of the 30-week observation period, suggesting regrowth of M. ulcerans. In contrast, 50% of the mice treated with the RIF؉AMK combination exhibited no lesion even after 30 weeks, suggesting cure. The remaining regimens could be assigned to one of three groups: (i) no activity (50 mg of AZM, 100 mg of AZM thrice weekly, TLM, and LVX); (ii) bacteriostatic activity, i.e., a delay in footpad swelling shorter than the 4-week treatment duration (100 mg of AZM daily or once weekly, CLR thrice or once weekly, and MOX); or (iii) weak bactericidal activity (CLR daily and SPX). The RIF؉AMK combination and possibly RIF؉STR warrant further study for the treatment of M. ulcerans infection in humans.Mycobacterium ulcerans, the only toxin-producing mycobacterial species (5) is responsible for Buruli ulcer (BU), the third most prevalent mycobacterial disease throughout the world (20). Over the past 10 years, an increasing incidence of BU has been reported in several West African countries (6, 8) and Australia (4) and is now reported in most of sub-Saharan Africa, Central and South America, India, Southeast Asia, and Papua New Guinea (20).In vitro, M. ulcerans is susceptible to several antimicrobials, including rifampin (RIF), clarithromycin (CLR), streptomycin (STR), amikacin (AMK), sparfloxacin (SPX), and clofazimine (1,11,13,14,19). However, the clinical response of humans with BU to various antimicrobial regimens containing these drugs have not confirmed in vitro studies so that, to date, BU treatment still relies on extensive surgical excision of the lesions followed by skin grafting (20).In a previous study, the mouse footpad model of Fenner (3) was used to assess the in vivo activity of several drugs against M. ulcerans (2). CLR, for which in vitro activity against M. ulcerans has been demonstrated (11), exhibited only bacteri...