Our experiment aimed to evaluate the influence of extracts of white and black truffle mushrooms on human spermatozoa. The study utilized 28 samples of wild truffle fruiting bodies. In the experiment, we used ejaculate from male volunteers of active reproductive age (N=10, 25–35 years old). During the experiment, we assessed sixteen physiological parameters. Research has shown that extracts from black and white truffles related toTubersp. have a stimulating effect on spermatozoa. The average path sperm velocity, curvilinear velocity and beat cross-frequency were increased by 56%, 48% and 50% respectively. Linearity showed a significant increase by 56% and straightness by 48%. This could be useful in the development of drugs to enhance sperm activity and lifespan. Additionally, extracts from black truffles have been found to have negative effects on spermatozoa, which could be relevant for developing new contraceptive drugs. Our study demonstrated, for the first time, the influence of methanol extracts ofTubersp. mushrooms on male gametesin vitro.