Resistance to macrolides has increasingly been reported for Group A streptococci. In this study, the in vitro antibiotic susceptibility pattern of 305 clinical isolates of S. pyogenes was determined. Strains were isolated during 1996 from pharyngeal swabs of children with uncomplicated pharyngitis living in 2 Italian cities: Milano and Catania, situated in the North and South of Italy, respectively. All isolates were found to be fully susceptible to penicillin and other beta-lactam agents tested. Susceptibility to macrolides differed markedly between the two centers with relatively high resistance rates to erythromycin being observed in Milano (30%) as compared to Catania (3%). Resistance to erythromycin was always crossed with that of the other 14- and 15-membered macrolides tested. However, resistance to josamycin and clindamycin was generally found in approximately 25% of the erythromycin-resistant (ER) strains. The erythromycin-resistant isolates from Milano and Catania (58 strains) were further subdivided into the three previously described resistance phenotypes: constitutive, inducible, and novel resistance phenotypes. The novel resistance phenotype accounted for 58% of all resistant strains, while 17% and 26% were found to be of the inducible and constitutive resistance phenotypes. Strains of the novel resistance phenotype were characterized by lower MIC values (MIC90 = 16 mg/L) to 14 and 15 carbon atom macrolides as compared to the other two phenotypes (MIC90 > 128 mg/L), and retained susceptibility to clindamycin and to josamycin, a 16 carbon atom macrolide. Resistance to tetracyclines was found in 25% to 36% of the ER isolates as compared to 2% to 10% of the susceptible strains. In particular, resistance to this agent was more commonly associated to isolates belonging to the novel and constitutive resistance phenotypes. MIC values for chloramphenicol in all isolates were within the susceptible or intermediate range; decreased susceptibility to this agent did not appear to be associated with erythromycin resistance.