Propolis is a product that is rich in phenolic compounds and can be utilized in animal nutrition as a dietary additive. In this study, the effects of a Brazilian green propolis extract on rumen fermentation and gas production were determined. The fate of propolis phenolic compounds in the rumen medium was also investigated. Fermentation was done in 24-h batches over three periods. Inoculates were obtained from cows fed on grassland hay and concentrate. Propolis extract in a hydroalcoholic solution was applied at increasing doses to the substrate (1 to 56 g/kg). The fermentation substrate consisted on a mixture of alfalfa hay, soybean meal, and wheat grain mixture in dry matter. After 24 h of fermentation, seven new compounds were observed in the medium in amounts that correlated to the propolis dose. The dose of propolis extract linearly decreased the pH of the medium and linearly increased propionate production, which reduced the acetate-to-propionate ratio and influenced the total production of short-chain fatty acids. Propolis also linearly reduced methane production and increased the carbon dioxide-to-methane ratio. Ammonia nitrogen levels and in vitro digestibility of organic matter were similar among the treatments. The combination of increased propionate production and decreased methane production suggests better energy utilization from the feed.