We examined antibacterial activities of 4 kinds of macrolides, erythromycin (EM), clarithromycin (CAM), azithromycin (AZM) and rokitamycin (RKM), against 6 bacterial species of clinical strains isoleted in 2002. Bacterial isolates used were each 50 strains of methicillin-susceptible Staphylococcus aureus (MSSA), Streptococcus pyogenes, Streptococcus agalactiae, Moraxella (Branhamella) catarrhalis, Haemophilus influenzae and 43 strains of Streptococcus pneumoniae. S. agalactiae were derived from gynecological samples, and other species were isolated from respiratory specimens. Antimicrobial activities against S. aureus, S. pyogenes, S. agalactiae, M. catarrhalis and H. influenzae of 14-membered macrolides, such as EM and CAM, were higher than those of 16-membered macrolide, RKM. By contrast, against S. pneumoniae, RKM was more effective than 14-membered macrolides. Six, three and four strains of S. aureus, S. pyogenes and S. agalactiae, respectively, were resistant to macrolides. Thirty-five among 43 pneumococcal isolates were resistant, and 15 of the 35 were highly-resistant, MIC of > 128 micrograms/ml, to any one of EM, CAM or AZM. Isolation frequency of resistant strains to RKM was lower than those to 14- and 15-membered macrolides: only one strain was highly-resistant and 12 were intermediately-resistant. No resistant strain was recognized in M. catarrhalis and H. influenzae. Further, we analyzed the resistant mechanisms, methylation or efflux, of macrolide resistant strains by the double-disk method. Methylation was major mechanism in S. aureus, and in S. pyogenes, all of the resistance was caused by methylation. In S. agalactiae and S. pneumoniae, methylation and efflux shared about half and half.