Abstract
In the preparation of gonococcus polysaccharides, we have encountered much difficulty in growing this organism in large quantities. While several liquid and semisolid media 1 2 3 4 have been used to grow this organism in large amounts and to keep its viability for a limited period, none was found to serve our purpose satisfactorily. The best medium is probably that of Singh 5 but its preparation is rather complicated. In the course of our investigation, we have found the following medium to be simple and efficient, not only for mass-production of gonococcus but also for preserving its viability for a long period.
The medium was prepared by simply mixing 4 parts of Tyrode solution and one part of horse or human serum. When properly prepared this mixture always gives a pH of 7.8. It was sterilized by filtration through Seitz filter, after which it was put into sterile flasks plugged with sterile rubber stoppers and finally sealed with paraffin before incubation at 37°C for 48 hours to ensure sterility.
The ability of this medium to support the growth of gonococcus was tested with 7 freshly isolated strains and one stock strain of gonococcus. Following inoculation with a suspension of gonococcus derived from these strains and upon incubation at 37°C diffuse granular growth began to appear after 48 hours. On further incubation a considerable portion of the growth appeared to have settled down at the bottom of the flasks. The maximal growth, however, was not reached until the end of a week's incubation. The growths of 7–10 days' incubation at 37°C were found to be rich enough to yield approximately 1 mg of crude gonococcus polysaccharides per 100 cc of the culture. In our experience a similar yield of gonococcal polysaccharides would require the growth from 15 petri dishes (10 cm in diameter) of nutrient agar pH 7.4 containing 5% horse blood.
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