The objective of this study was to evaluate the aerobic rumen mycobiota from three age groups of Nelore beef cattle reared extensively on lignified pasture. The experiment was randomized and sampled 50 steers, 50 cows, and 50 calves grazed on Brachiaria spp. pasture during the dry season. Rumen fluid in all animals was aromatic, slightly viscous, and greenish-brown in color. Microscopic examination revealed monocentric and polycentric anaerobic fungi in similar proportions (P>0.05) in the rumen fluid of cows and steers. However, these microorganisms were not identified in any of the samples from calves. In culture exams, aerobic filamentous population was significantly higher for rumen fluid of cows compared to the other two groups. Microculture and rDNA sequence analyses showed Aspergillus spp. as the most frequent aerobic fungus among the isolates from the three bovine groups evaluated. Biochemical profiles were determined by the growth level of yeast isolates with 44 nutrient sources. Ten different yeast profiles were obtained, and yeast isolates from cow ruminal fluid showed ability to catabolize greater diversity of carbon and nitrogen sources. The differences in the fungal populations observed in this study could be explained by microbial and physiological interactions existing in the ruminal ecosystem of each age bovine group. The present study showed the fungal population of the rumen related with differences among age of cattle raised in lignified pastures. Metabolic capabilities of mycelial fungi or yeast identified in this study may be employed in new probiotics or microbial additives for different bovine categories.