FIG 1.

Experimental design. Mice were vaginally inoculated with C. muridarum (labeled “CM” in the figure) daily for three consecutive days. Viable C. muridarum shedding and recovery of viable N. gonorrhoeae (labeled “NG” in the figure) were monitored by regular vaginal swabbing, every 2 to 3 days, for the duration of the experiment. During chlamydial latency, vaginal smears were collected to determine estrous cycle stage, and selected mice (anestrus and diestrus stage) were treated with subcutaneous (labeled “SC” in the figure) injections of Premarin and intraperitoneal (labeled “IP” in the figure) injections of antibiotics (vancomycin hydrochloride and streptomycin sulfate [Vanc/Strep]) for 4 days, as shown. Two days after initiation of Premarin/antibiotics, mice were vaginally inoculated once with N. gonorrhoeae. After the cessation of antibiotic injections, Strep was supplied in drinking water. Mice that were not selected for Premarin/antibiotics treatment were sacrificed at day 23 (early sacrifice), and all other mice were sacrificed at day 36 (late sacrifice). At sacrifice, rectal swabs were collected for viable C. muridarum enumeration, and necropsy was performed to evaluate gross genital tract pathology; genital and intestinal tracts tissues were collected for evaluation of pathology.