Modeling Murine Vaginal Colonization by Anaerobic Bacteria

Take a female mouse treated with a hormone that conditions the vaginal environment, increasing its susceptibility to infection.

Lift the mouse by its tail while it grips the cage with its forepaws to elevate the hind quarters and expose the vaginal opening.

Rinse the vaginal lumen with an anaerobic buffer to flush out mucus, a protective barrier against pathogens, along with dead epithelial cells, and endogenous vaginal microbes.

Immediately inoculate the vagina with a suspension of an anaerobic bacterial pathogen, while keeping the mouse restrained to minimize fluid loss.

For co-inoculation, administer a suspension of another anaerobic bacterial pathogen using the same volume as before.

House the inoculated mouse separately from uninoculated mice to prevent cross-contamination.

Following inoculation, the bacteria adhere to the vaginal epithelium, grow into colonies, and develop a protective biofilm layer that acts as a barrier against host immune responses.

Prepare the vaginal lavage tubes in the anaerobic chamber by pipetting 70 microliters of sterile anaerobic PBS into 1.5-milliliter microcentrifuge tubes labeled with mouse numbers.

Close the tubes tightly and take them out of the chamb