PCR based genetic analysis of Aeromonas hydrophila isolated from the aquaculture systems.

Divya, P R (2002) PCR based genetic analysis of Aeromonas hydrophila isolated from the aquaculture systems. Masters thesis, Central Institute of Fisheries Education.

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    Abstract

    Molecular genetic characterization of Aeromonas hydrophila isolates from aquaculture systems was carried out to evaluate the genetic heterogeneity and phylogenetic relationship among them, using PCR based RAPD technique. This technique was found to have many advantages like being simple, easy, rapid and highly discriminatory between isolates within the species. PCR amplification of the DNA from the bacterial isolates using five random primers produced 46 amplicons, which were distinct, as scorable bands on agarose gel electrophoresis. Polymorphism of RAPD profile was evident with a unique pattern for each isolate. The level of polymorphism among the amplified fragment was 95% indicating genomic heterogeneity among the isolates within the species of A.hydrophila, which were homogenous phenotypically. Choice of the primer was found to be important as the discriminatory ability of RAPD varied with the primer used for amplification. Two of the amplicons shared by all the isolates were identified which could serve as species specific markers. Cellular protein profile of the isolates generated through SDS-PAGE did not reveal any polymorphism as all the isolates presented uniform pattern indicating its usefulness as a tool for species level identification. Coefficients of genetic identities between the isolates were generally high compared to the genetic distances. Phylogenetic tree depicting the relative genetic relationship among the isolates was also made. RAPD analysis was found to be a desirable tool for molecular genetic characterization and for evaluation of genetic heterogeneity in A.hydrophila, which remain hidden by other techniques like morphological and biochemical characterization and cellular protein profile.

    Item Type: Thesis (Masters)
    Uncontrolled Keywords: Genetic analysis; Aeromonas hydrophila; Aquaculture systems
    Subjects: Dissertations
    Fish Diseases
    Divisions: CMFRI-Kochi > Mariculture
    Subject Area > CMFRI > CMFRI-Kochi > Mariculture
    CMFRI-Kochi > Mariculture
    Subject Area > CMFRI-Kochi > Mariculture
    Depositing User: Users 17220 not found.
    Date Deposited: 06 Sep 2016 10:36
    Last Modified: 07 Oct 2016 04:22
    URI: http://eprints.cmfri.org.in/id/eprint/11056

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