The ability of eighteen isolated ruminal fungi to degrade highly fibrous substrates was evaluated by fermentations lasting 120 hours. The graminea Bothriochloa pertusa was used as the unique carbon source. The experiment involved isolates belonging to the Neocallimastix, Orpinomyces and Piromyces genera. According to the Gompertz model, the establishment phase of the isolated fungi was on average 23,8 h ± 4.20, and reached a gas production level of 195 ml/g DM ± 7,52. The half gas production was achieved between 35 and 55 hours and varied with each ruminal fungi. The highest value of the maximum gas production rate from all the isolated fungi was 5,7 ml/h; this was produced by an isolated of Orpinomyces genus. The degradability of dry matter (DM) and neutral detergent fibre (NDF) of al the isolated fungi was on average 58% ± 2 and 46 % ± 2, respectively. With regard to volatile fatty acids (VFA) production all of the isolates were on average 22,04 mM ± 1,5; with an acetic acid proportion of 93% ± 0,01. The highest values of fibrolytic activity for carboxymetyl cellulase, microcrystalline cellulase and xylase were 9,56, 1,39, 303,12 international units (UI), respectively; this confirmed preferences toward hemi-cellulosic substrates by ruminal fungi. Five isolated fungi with the most diversity in the evaluated parameters, principally in terms of degradability of DM, NDF and enzymatic capacity were selected by principal component analysis (PCA), followed by grouping clusters. These isolates were one Neocallimastix, two Orpinomyces and two Piromyces genera.