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Rearing and Injection of Manduca sexta Larvae to Assess Bacterial Virulence
Published on: December 11, 2012
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Enterococcus faecalis 6-phosphogluconolactonase is required for both commensal and pathogenic interactions with
Jonathan F Holt1, Megan R Kiedrowski2, Kristi L Frank3
1Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin, USA Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut, USA.
Infection and Immunity
|November 12, 2014
Summary
This study identifies key Enterococcus faecalis genes, including pglA, crucial for gut persistence and pathogenesis in Manduca sexta. These findings offer insights into bacterial host interactions and potential infection targets.
Area of Science:
- Microbiology
- Insect Pathology
- Bacterial Genetics
Background:
- Enterococcus faecalis transitions between commensal and pathogenic roles in hosts.
- Understanding E. faecalis gut colonization factors is crucial for controlling infections.
Purpose of the Study:
- Identify E. faecalis genes essential for gut commensalism in Manduca sexta.
- Investigate the role of identified genes in pathogenesis and persistence.
Main Methods:
- Utilized recombinase-based in vivo expression technology (RIVET) to screen for upregulated genes in the M. sexta gut.
- Generated gene deletion mutants for functional analysis.
- Performed in vitro biofilm assays and in vivo pathogenesis/persistence studies.
Main Results:
- Identified 50 upregulated E. faecalis genes in the M. sexta gut, including the 6-phosphogluconolactonase gene pglA.
- A pglA deletion mutant showed impaired pathogenesis and gut persistence, with enhanced biofilm formation.
- Mutations in four other identified genes impaired pathogenesis but not gut persistence.
Conclusions:
- This study provides the first identification of genetic determinants for E. faecalis interactions with M. sexta.
- Identified bacterial factors may inform strategies for preventing E. faecalis infections in various host-associated microbial communities.
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