Presumptive Identification, Cultural Characterisation and Collagenase Production from Clostridium novyi-NT

Abstract

Clostridia are well known for their ability to excrete varieties of enzymes into the environment. Clostridia sp. being medically important organisms produces a wide variety of extra-cellular toxins including proteases like collagenases. C. novyi-NT is medically and therapeutically important organisms since endospores of which are being employed in oncolytic therapy as well as immunomodulatory therapy. The aim of this work was to identify and investigation of cultural characters of C. novyi-NT after its growth. In routine presumptive tests, the vegetative culture shows dual and bias Gram nature behaviour as young culture appeared as Gram positive while the old culture shows Gram negative characters. The endospores appeared as green ellipse within and outside of cells while the vegetative cells as pinkish red with terminal or subterminal shape. The colonies appeared with flat, circular and greyish in colour, surrounded by zone of hemolysis on blood agar media at 35-37°C for 48 hrs. The bacteria observed “swarms” as a continuous film over the entire plate, with ?-haemolysis pattern. C. novyi- NT is unable to produce lethal a-toxin and hence does not produce lecithinase to give negative Nagler’s reaction. This bacterium is capable of producing the exoenzyme lipase to give clear zone surrounding its growth in tributyrin agar. C. novyi- NT gives negative indole and urease test. C. novyi- NT had shown its collagenolytic activity on the collagen agar plates. The collagenase presence was detected by the clear zone formation surrounding zizag growth of bacteria.