Abstract: A total of 115 Pasteurella multocida isolates were collected from various sources of animals in Malaysia and were purified by Veterinary Diagnostic Laboratory, Petaling Jaya. Isolates were subcultured once on BHI blood agar, incubated at 37?C overnight before amplified in BHI broth. Before carrying out to any further studies, all the isolates were first reconfirmed as Pasteurella multocida by gram staining, colonial morphology and a series of biochemical tests. The cultures of Pasteurella multocida were tested for the production of hyaluronidase by using modification method of Smith and Willett (1968). Hyaluronidase production was demonstrated in all isolates of Pasteurella multocida serotype B, whereas, none was detected in serotype A, D and untypable.