The possible inhibition of Neisseria gonorrhoeae on modified Thayer-Martin (VCNT) medium was investigated by inoculation of multiple media with specimens from 3,490 patients. N. gonorrhoeae was recovered from 461 patients, and in 24 cases (5.2%) it was isolated on drug-free medium only; 18% of the recoveries were on VCNT medium only. The minimal inhibitory concentration (MIC) of benzylpenicillin was determined for 411 of the strains, and 175 were examined for responses to 12 other antibiotics and for auxotype. Of the 24 strains isolated on drug-free medium only, one was inhibited by trimethoprim at a concentration of 2.0 micrograms/ml and four others had MICs of vancomycin of less than or equal to 2.0 micrograms/ml. The remainder were resistant to vancomycin, trimethoprim, and colistin at the concentrations present in VCNT medium. Unexpectedly, four strains isolated on both VCNT and drug-free medium had MICs of vancomycin of less than or equal to 3.0 micrograms/ml and were defined as hypersusceptible. Genetic tests showed that gonococci resistant to less than or equal to 0.5 microgram of vancomycin/ml differed genotypically from those resistant to 1.0 microgram/ml. The eight strains hypersusceptible to vancomycin were highly susceptible to various other antibiotics. Their nutritional requirements included hypoxanthine (Hyx-) and uracil (Ura-), and all but one also required arginine (Arg-), which for two strains could not be replaced by ornithine (Arg0-). Pro-,Arg0-,Ura- (5.7%) and Arg0-,Ura- (1.1%) auxotypes were found at this time but not in earlier studies of gonococci isolated in the same clinic.