Plasmid DNA analysis of pasteurella multocida Serotype B isolated from Haemorrhagic Septicaemia outbreaks in Malaysia

Jamal, H.; Chua, K.H.; Frederick, D.; Mahmood Ameen, A.; Salmah, I. (2005) Plasmid DNA analysis of pasteurella multocida Serotype B isolated from Haemorrhagic Septicaemia outbreaks in Malaysia. Malaysian Journal of Microbiology, 1 (2). pp. 35-39. ISSN 1823-8262

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    Abstract

    A total of 150 purified isolates of Pasteurella multocida serotype B were used (Salmah, 2004) for plasmid DNA curing experiment to determine hyaluronidase activity, antibiotic resistance pattern (ARP) and mice lethality test (LD50) for their role of pathogenicity. A plasmid curing experiment was carried out by using the intercalating agent; ethidium bromide and rifampicin, where it was found all the plasmids had been cured (plasmidless) from Pasteurella multocida. All of these plasmidless isolates maintained their phenotypic characteristics. They showed the same antibiotic resistancepattern as before curing, produced hyaluronidase and possessed lethality activity in mice when injected intraperitoneally(i.p). Based on this observation, the antibiotic resistance, hyaluronidase activity and mice virulence could probably be chromosomal-mediated. Plasmids were detected 100 in all P. multocida isolates with identical profile of 2 plasmids size 3.0 and 5.5 kb. No large plasmids could be detected in all isolates. Since all the isolates appeared to have identicalplasmid profiles, they were subjected to restriction enzyme(RE) analysis. From RE analysis results obtained, it can be concluded that the plasmid DNA in serotype B isolates are identical. Only 4 of 32 REs were found to cleave these plasmids with identical restriction fingerprints; BglII, HaeIII, RsaI and SspI. From RE analysis results, it can be concluded that the plasmid DNA isolates are identical. This plasmid might not played any role in pathogenicity of Pasteurella multocida serotype B, however this information is important for the construction of shuttle vectors in genetic studies of the pathogenicity of haemorrhagic septicaemia(HS).

    Item Type: Article
    Journal or Publication Title: Malaysian Journal of Microbiology
    Additional Information: Department of Molecular Medicine, Faculty of Medicine Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA
    Uncontrolled Keywords: Pasteurella multocida serotype B, plasmid, haemorrhagic septicaemia, hyaluronidase, pathogenicity.
    Subjects: R Medicine
    Divisions: Faculty of Medicine
    Depositing User: Ms azrahani halim
    Date Deposited: 07 Nov 2012 10:28
    Last Modified: 07 Nov 2012 10:28
    URI: http://eprints.um.edu.my/id/eprint/3843

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