OBJECTIVE To explore a new method of preventing Neisseria gonorrhea (N. gonorrhoeae) by simulating intercourse in mice and verify the effect of a long-acting antibacterial material in the gonococcal infection in mice. METHODS (1) Simulated male-female gonococcal transmission: the estradiol-treated BALB/c female mice were inoculated with N. gonorrhoeae WHO-L via a vaginal route. In treated group, the mice received the long-acting antibacterial material via vaginal route while deionized water was administered in the control group. At Days 2 - 10 post-inoculation, the average number of polymorphonuclear cells (PMNs) was monitored in stained smears of vaginal cells from test mice. And the vaginal fluid was cultured for N. gonorrhoeae at Days 2, 4, 5 post-inoculation. N. gonorrhoeae genital tract infection was determined to compare the difference between the two groups. (2) Simulated female-male gonococcal transmission: a treated or sham glass rod was used instead of male mice's penis to simulate intercourse in gonococcus-infected female mice. The vaginal fluid on top of glass top was eluted and cultured for N. gonorrhoeae and the difference of relevance ratios compared. RESULTS (1) In the experiment of simulated male-female gonococcal transmission, At Days 4 and 5, the positive rates of culture for N. gonorrhoeae of the tested group were lower than those of the control group, and there was significant difference between both groups (10.0% vs 57.5%, 7.5% vs 45.0%, both P < 0.01). The percentage of PMNs in the treated group was significantly lower than that of the control group at Days 5 - 8 post-inoculation (all P < 0.05). (2) In the experiment of simulated female-male gonococcal transmission, the positive rates of culture for N. gonorrhoeae from the vaginal fluid on top of glass top were 8.3% and 0 at Days 1 and 2 in the tested group respectively. And they were significantly lower than those of the control group (72.7% and 45.5% respectively, both P < 0.01). CONCLUSIONS After topical application in murine vagina, the long-acting antibacterial material forms a layer of physically antibacterial molecular film to prevent the occurrence of N. gonorrhoeae genital tract infection or colonization in mice. It may be used as a liquid condom for male mice.