Einfache Methode zum schnellen Nachweis bakterieller Hyaluronidase in K-Hyaluronat-haltigem Gel

Abstract

Summary For detection of hyaluronidase activities we investigated several groups of bacteria. The bacteria were inoculated on a 1,5% agarose gel in Petri plates of 4 cm ∅ or gel discs of 7 mm ∅, containing 0,1 % of K-hyaluronate as well as nutritient medium, and were incubated for 2–20 h at 37°C in a moist chamber. Subsequently some ml of a 10% solution of cetylpyridiniumchloride were poured on the gel to precipitate the polymere hyaluronate. If the hyaluronate was depolymerized by hyaluronidase, a translucent area was visible around the colonies. We found out, that a gel layer of 1 mm was sufficient to detect the small amounts of hyaluronidase, which were produced by bacteria within an incubation time of 2 h. These results were confirmed by incubation for 20 h and in some cases 36 h. The hyaluronidase production by different anaerobic Clostridium strains was always proved after a 20 h growth period. The bacteria were inoculated with the whole loop of a self made platin sowing wire loop. By this method quantitative differences of hyaluronidase activities between different strains of bacteria could be detected.