Presence of hydrogen peroxide in media used for cultivation of Neisseria gonorrhoeae

Abstract

Defined complex media used for cultivation of Neisseria gonorrhoeae were tested for the presence of H2O2 by both a spectrophotometric and a polarographic assay. H2O2 (35 to 165 microM) was present in all media tested. In the defined media, H2O2 was generated by the interaction of cysteine with other amino acids. The addition of the chelator 8-hydroxyquinoline prevented formation of detectable H2O2, suggesting that metal ions were necessary. The persistence of H2O2 varied greatly among different media. Medium components which affected the presence of H2O2 were pyruvate, oxalacetate, and sodium sulfite. Sodium sulfite also generated superoxide radical. In liquid medium containing H2O2, the endogenous gonococcal catalase present in an inoculum of about 2 X 10(7) colony-forming units/ml destroyed detectable H2O2. The long lag phase which resulted from a 10-fold lower inoculum could not be shortened by the addition of exogenous catalase. Small amounts of residual H2O2 in agar plates of complex medium affected the viability of gonococci which had been suspended in buffer and incubated for 60 min at 37 degrees C. Addition of pyruvate or catalase increased viable counts in medium containing H2O2.