Intensive silvopastoral systems are being increasingly disseminated in the tropical areas of Mexico, Central and South America, so it is important to know the quality of the animal products generated in these systems. Fatty acid profile of fresh soft cheese (Domiati type) produced from milk of cows grazing Leucaena leucocephala associated with Cynodon nlemfuensis in an intensive silvopastoral system (ISS) was evaluated and compared with a cheese produced from cows grazing a traditional monoculture system (MS) of C. nlemfuensis. The legume L. leucocephala contains phenolic compounds that could affect the fatty acid biohydrogenation in the rumen and therefore the fatty acid profile of resultant milk and cheese. Milk samples from cows in both diets (systems) were processed monthly for cheese making and cheese samples were analyzed for fatty acid profile. The experimental period lasted for 10 months covering the rainy and dry season of a tropical area in Mexico. The results showed that the cheese produced in ISS had significantly lower content (P<0.05) of the saturated fatty acids lauric, myristic and palmitic and significantly higher content (P<0.05) of stearic, linoleic and γ-linolenic acids. Additionally, higher content (P<0.05) of the polyunsaturated fatty acids and omega-6 fatty acids were found in the cheese from ISS. The results suggest that fresh soft cheese produced from silvopastoral systems based on L. leucocephala has a better fatty acid profile than that produced from the traditional MS with grass only.