The in vitro activities of GAR-936, the 9-t-butylglycylamido derivative of minocycline, were compared with those of doxycycline, minocycline, and tetracycline against 527 gram-positive clinical isolates. GAR-936 inhibited all strains, including those resistant to other tetracyclines, at concentrations of <2 g/ml, except two strains of JK diphtheroids for which the MIC was 4 g/ml.Although tetracyclines remain valuable therapeutic agents for a variety of infections, resistance to this class limits their use against many important gram-positive bacterial pathogens. For example, only 33% of enterococci recovered at our hospital during 1997 to 1999 were susceptible to tetracycline. The glycylcyclines are novel tetracycline analogs that have activity against organisms resistant to older compounds of this class (5,14). They inhibit protein synthesis on wild-type ribosomes and on TetM-protected, tetracycline-resistant ribosomes (15). These compounds also inhibit organisms with tetracycline efflux determinants (14). GAR-936, the 9-t-butylglycylamido derivative of minocycline, appears to be both better tolerated by hosts and more active against tetracycline-resistant strains than earlier glycylcyclines (14). The present study examined the in vitro activities of GAR-936 against gram-positive bacteria, including strains resistant to -lactams, glycopeptides, and other tetracyclines.Routine clinical isolates collected at the Beth Israel Deaconess Medical Center were included regardless of tetracycline resistance patterns. Staphylococci and most pneumococci were recovered in 1998. Strains with unusual resistance traits, including glycopeptide-resistant or -lactamase-producing enterococci and -lactam-resistant streptococci, had been referred to our laboratory from various sources over several years (2,4,16). Wyeth-Ayerst Laboratories, Pearl River, N.Y., kindly provided GAR-936. Tetracycline, minocycline, and doxycycline were purchased from Sigma Chemical Company, St. Louis, Mo.Activities of the antimicrobials were determined by agar dilution methods on Mueller-Hinton II agar (12). Media were supplemented with 5% sheep blood for testing streptococci and corynebacteria. Inocula of approximately 10 4 CFU were applied to the surfaces of plates and were incubated for 16 to 20 h at 35°C in air or in 5% CO 2 (for Lactobacillus spp., Leuconostoc spp., Streptococcus spp., Pediococcus spp., and diphtheroids). Pneumococci were also tested by broth microdilution (12).The results of susceptibility studies are shown in Table 1. GAR-936 was four times less active than minocycline against oxacillin-susceptible and -resistant strains of Staphylococcus aureus, according to a comparison of MICs at which 90% of the isolates tested were inhibited (MIC 90 s). However, isolates intermediately susceptible or resistant to the other compounds were inhibited by GAR-936 at Յ1 g/ml. GAR-936 inhibited two strains of vancomycin-intermediate S. aureus (resistant to tetracycline and minocycline) at concentrations of 1 and 2 g/ml.Against coagulase-negative s...