A medium, solidified with gellan gum, for determining the Na+ requirement of Vibrio species

Abstract

Until now there has not been a satisfactory solid medium for determining the growth responses, to Na+, of marine and other bacteria that have specific growth requirements for Na+. A solid medium would be useful to investigators who would like to take advantage of the efficiency of multipoint inoculation when testing for a Na+ requirement. By using 1% gellan gum (Gel-GroTM) as the solidifying agent a medium was formulated that had a contaminating level of Na+ of slightly less than 2 mM in the basal medium. Two species of Aeromonas, which do not require Na+ for growth, and 31 species of Vibrio, which require Na+, were tested for their growth responses to Na+ on this medium. The Aeromonas strains grew well, within 24 h, at all of the Na+ concentrations tested. Approximately 75% of the Vibrio strains did not grow on the basal medium even after a prolonged incubation period. The remaining species were able to grow on the basal medium, but not without a lag period. These lag periods were as short as 36 h for two of the species and in some instances as long as 312 h. These lag periods were of sufficient duration to determine that Na+ stimulated the growth of the Vibrio strains that were able to grow on the basal medium. Approximately 75% of the strains, representing most species of Vibrio, were able to grow if as little as 25 mM Na+ was present in the medium.Key words: low-sodium medium, Na+ requirement, gellan gum, agar substitute, marine bacteria.