Neisseria gonorrhoeae is a human pathogen of mucosal surfaces, thus laboratory manipulations must include appropriate safety measures. The growth requirements and behavior of the gonococcus are significantly different from many bacteria, necessitating modifications of common laboratory techniques. A fastidious organism, N. gonorrhoeae requires enriched media in a CO2 atmosphere at 35 degrees to 37 degrees C for growth. In addition, N. gonorrhoeae expresses potent autolysins whose activity increases following glucose depletion during stationary phase, leading to cell death. Long believed to be an obligate aerobe, the gonococcus is capable of anaerobic growth when provided with a suitable electron acceptor. This unit provides information for both aerobic and anaerobic growth, basic long-term and daily maintenance of gonococcal cultures, as well as safety considerations for laboratory studies.