Genome-wide mutagenesis identifies factors involved in enterococcus faecalis vaginal adherence and persistence

Norhan Alhajjar, Anushila Chatterjee, Brady L. Spencer, Lindsey R. Burcham, Julia L.E. Willett, Gary M. Dunny, Breck A. Duerkop, Kelly S. Doran

Research output: Contribution to journalArticlepeer-review


Enterococcus faecalis is a Gram-positive commensal bacterium native to the gastrointestinal tract and an opportunistic pathogen of increasing clinical concern. E. faecalis also colonizes the female reproductive tract, and reports suggest vaginal colonization increases following antibiotic treatment or in patients with aerobic vaginitis. Currently, little is known about specific factors that promote E. faecalis vaginal colonization and subsequent infection. We modified an established mouse vaginal colonization model to explore E. faecalis vaginal carriage and demonstrate that both vancomycin-resistant and -sensitive strains colonize the murine vaginal tract. Following vaginal colonization, we observed E. faecalis in vaginal, cervical, and uterine tissue. A mutant lacking endocarditis- and biofilm-associated pili (Ebp) exhibited a decreased ability to associate with human vaginal and cervical cells in vitro but did not contribute to colonization in vivo. Thus, we screened a low-complexity transposon (Tn) mutant library to identify novel genes important for E. faecalis colonization and persistence in the vaginal tract. This screen revealed 383 mutants that were underrepresented during vaginal colonization at 1, 5, and 8 days postinoculation compared to growth in culture medium. We confirmed that mutants deficient in ethanolamine catabolism or in the type VII secretion system were attenuated in persisting during vaginal colonization. These results reveal the complex nature of vaginal colonization and suggest that multiple factors contribute to E. faecalis persistence in the reproductive tract.

Original languageEnglish (US)
Article numbere00270
JournalInfection and immunity
Issue number10
StatePublished - Oct 2020

Bibliographical note

Funding Information:
This study was supported by NIH 5T32AI007405-28 to B.L.S., NIH/NIAID R21 AI130857 to K.S.D., NIH/NIAID R01 AI141479 to B.A.D., NIH/NIAID R01 AI122742 to G.M.D., American Heart Association grant 19POST34450124/Julia Willett/2018 to J.L.E.W., and a University of Colorado School of Medicine IMPA to K.S.D. and B.A.D.

Publisher Copyright:
Copyright © 2020 American Society for Microbiology.


  • Colonization
  • E. faecalis
  • Enterococcus
  • Ethanolamine catabolism
  • T7SS
  • TnSeq
  • Vaginal tract

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