Dirithromycin is a new macrolide antibiotic with an active metabolite, erythromycylamine. We evaluated the in vitro activities of both drugs against 16 isolates of Chiamydia trachomatis and compared them with that of doxycycline. In vitro susceptibility testing was performed with McCoy cell monolayers. The MIC was defined as the lowest concentration of antibiotic without inclusions. The MBC was defined as the lowest concentration of antibiotic yielding no inclusions after passage onto 24-h-old antibiotic-free McCoy cell monolayers. Dirithromycin and erythromycylamine appeared to be equally effective against these 16 strains of C. trachomatis (MIC for 90%o of strains tested, mg/ml; MBC for 90% of strains tested, 2 p,g/ml). Both were less active than doxycycline (MIC for 90%o of strains tested, 0.06 ,g/ml; MBC for 90%o of strains tested, 0.12 ,ug/ml). The combination of dirithromycin and erythromycylamine appeared to be additive.Dirithromycin is a new macrolide antibiotic which is metabolized to an active compound, erythromycylamine (2). It has in vitro activity comparable to that of erythromycin (9). Like erythromycin, dirithromycin is concentrated intracellularly (3). Erythromycin and other macrolides have been shown to have both in vitro and in vivo activity against Chlamydia trachomatis (1, 4, 5, 7). However, there is no published information on the activity of dirithromycin against C. trachomatis. Therefore, we tested the in vitro activities of dirithromycin and erythromycylamine against clinical isolates of C. trachomatis and compared them with that of doxycycline, an antibiotic commonly used to treat C. trachomatis infections. We also tested dirithromycin and erythromycylamine for synergy.Dirithromycin and erythromycylamine were obtained from Lilly Pharmaceuticals (Indianapolis, Ind.), and doxycycline was obtained from Sigma Chemicals (St. Louis, Mo.). These drugs were prepared as stock dilutions in concentrations of 1,280 ,ug/ml according to their stated potencies. Stock solutions were stored at -70°C for a maximum of 2 weeks. Drugs were diluted to the appropriate concentration with medium containing Hanks' balanced salt solution, amino acids, vitamins, 1% glutamine, 10% inactivated fetal calf serum, 5.4 g of glucose per liter, and 1 ,ug of cycloheximide per ml. Dilutions were made on the day of use. A total of 16 strains of C. trachomatis, including one serotype D strain (ATCC VR-885), one lymphogranuloma venereum type 2 strain (ATCC VR-902b), and 14 clinical isolates, were tested. All clinical isolates were of genital origin and had been passed fewer than 10 times in the laboratory. All isolates were passed an additional two times in antibiotic-free medium before susceptibility testing and were stored at -70°C until the time of use.The antimicrobial susceptibility of C. trachomatis was determined by using 96-well dilution plates as previously described (6). McCoy cell monolayers, 24-h old and grown in antibioticfree medium, were inoculated with a dilution of C. trachomatis test strain known to yield 500 ...