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    GENETIC ANALYSIS OF CLOSTRIDIUM PERFRINGENS: INSIGHT INTO EVOLUTION OF VIRULENCE

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    Author
    Sawires, Youhanna Sobhy
    Issue Date
    2005
    Keywords
    Clostridium perfringens
    MLVA
    Virulence
    Evolution
    Strain typing
    Epidemiology
    Committee Chair
    Songer, J. Glenn
    
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    Publisher
    The University of Arizona.
    Rights
    Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
    Abstract
    Clostridium perfringens is an important pathogen in veterinary and medical fields. Understanding epidemiology of C. perfringens diseases and evolution of virulence within C. perfringens necessitates an efficient, time and cost effective strain typing method. Multiple-locus variable-number tandem repeat analysis (MLVA) has been applied to typing of other pathogens and we describe here the development of a MLVA scheme for C. perfringens. We characterized five VNTR loci, and screened 112 C. perfringens isolates to evaluate typability, reproducibility, and discriminatory power of the scheme. All isolates were assigned a MLVA genotype and the technique has excellent reproducibility, with a numerical index of discrimination of 0.995. Thus, MLVA is an efficient tool for C. perfringens strain typing, and being PCR based makes it rapid, easy, and cost effective. In addition, it can be employed in epidemiological, ecological, and evolutionary investigations of the organism.Virulence of this species is not fully understood and it does seem that distribution of the toxin/enzyme genes is erratic within the population. We used the MLVA scheme to investigate evolution of virulence and population structure of this species. Analysis of the phylogenetic signal indicates that acquisition of the major toxin genes and other plasmid-borne toxin genes is a recent evolutionary event, and their maintenance is essentially a function of the selective advantage they confer to strains carrying them in certain micro-niches under different conditions. In addition, it indicates the ability of virulent strains to cause disease in different hosts. More interestingly, there is evidence that certain normal flora strains are virulent when they gain access to a different host species. Analysis of the population structure indicates that recombination events are the major tool that shapes the population and this panmixia is interrupted with frequent clonal expansion that mostly corresponds to disease processes. Signature of positive selection was detected in the alpha toxin gene, suggesting the possibility of adaptive alleles on the other chromosomally-encoded determinants. Finally, C. perfringens proved to have a dynamic population, and availability of more genome sequences, use of comparative proteomics and of animal models would provide more insight into the pathogenicity of this organism.
    Type
    text
    Electronic Dissertation
    Degree Name
    PhD
    Degree Level
    doctoral
    Degree Program
    Pathobiology
    Graduate College
    Degree Grantor
    University of Arizona
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