Determination of Shigella sonnei Virulence using Caenorhabditis elegans Infection Model

Introduction: Shigella is the second leading cause of diarrhoeal mortality, leading to more than 60 thousand deaths in children younger than five years. In vivo studies of Shigella were traditionally done on non-primate mammalians such as rabbits and guinea pigs with a long-life cycle. A soildwellin...

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Bibliographic Details
Published inAsia-Pacific journal of molecular biology and biotechnology Vol. 30; p. 52
Main Authors Wong, Bao Chi, Tan, Hock Siew
Format Journal Article
LanguageEnglish
Published Kuala Lumpur Malaysian Society for Molecular Biology and Biotechnology 01.06.2022
Subjects
Accumulation
Bacteria
Bacterial diseases
Bacterial infections
Caenorhabditis elegans
Colonization
Fatalities
Fluorescence
Fluorescence microscopy
Guinea pigs
In vivo methods and tests
Infections
Life cycles
Microscopy
Nematodes
Pathogenesis
Rabbits
Shigella
Shigella sonnei
Strains (organisms)
Virulence
Worms
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Summary:Introduction: Shigella is the second leading cause of diarrhoeal mortality, leading to more than 60 thousand deaths in children younger than five years. In vivo studies of Shigella were traditionally done on non-primate mammalians such as rabbits and guinea pigs with a long-life cycle. A soildwelling nematode Caenorhabditis elegans, however, is easy to handle and has a shorter life cycle. In this project we aim to demonstrate the usability of C. elegans for Shigella sonnei pathogenesis and virulence research. Methods: Age-synchronised C. elegans were exposed to different strains of S. sonnei. The most virulent S. sonnei strain was selected for further characterization. The bacterial colonization assay was done by plating out the bacteria present within the worms after infection. Fluorescence microscopy was also performed using the virulent S. sonnei strain tagged with RFP. Results: Two S. sonnei strains were tested in C. elegans, namely ATCC 29930 and ATCC 25922. Both ATCC strains were virulent towards the nematode. ATCC 29930 was the most virulent, causing 50% deaths within 5 days. Bacterial colonization assay showed the accumulation of live S. sonnei ATCC 29930 reaching its plateau after 72 hours. Fluorescence microscopy also showed that ATCC 29930 begins to accumulate in the nematode guts after 3 hours. Conclusion: The nematode C. elegans is a useful model to determine the virulence of bacteria as a preliminary screen. This can be done by determining the survival of nematode after bacterial infection, the quantification of bacterial accumulation within the worm, and fluorescence microscopy.
ISSN:0128-7451