Condensed tannins are polyphenolic compounds which can exert beneficial effects in ruminants. They have the ability to bind proteins and decrease their degradation. They have also been reported to reduce methanogenesis and improve ruminants performances. The present work aimed to study the effect of purified condensed tannins from Acacia horrida extract on fermentation parameters, gas and methane production. In this context, seven substrates were selected: four Acacia species (Acacia pycnantha, Acacia dealbata, Acacia horrida and Acacia cyanophylla) and three plant by-products (date palm leaves, grenade peel and artichoke stems). Biological activity of tannins was evaluated by the incubation of the substrates in vitro with Polyethylene glycol (PEG). Tannins decreased concentration of all fermentation parameters (gas (p < 0.0001), methane (CH4) (p= 0.0983), ammonia (N-NH3) (p= 0.0382), Volatile Fatty Acids (VFA) (p= 0.0009), acetate (p <0.0001), propionate (p= 0.1024), butyrate (p= 0.0373), isobutyrate (p= 0.0046), valerate (p < 0.0001), isovalerate (p= 0.0032)). Substrates were also characterized by their content in main nutrients (dry matter (DM), organic matter (OM), ash, crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) ,acid detergent lignin (ADL)) and in secondary metabolites (total phenols (TP), total tannins (TT) and condensed tannins (CT) ). All substrates presented high CP content except by-products (> 100 g/kg DM) and moderate cell-wall components. CT content was comprised between 120.5 and 680.4 g/kg DM in plants and between 23.1 and 170.4 g/kg DM in plant by-products. A. horrida presented the highest biological activity. Thus, their CT were extracted and purified on a sephadex LH-20 column. Purified tannins from A. horrida were incubated with two different forages alfalfa hay and barley straw at three different concentrations: 50, 100, 150 mg/g DM. Results showed that purified tannins from A. horrida had no effect on fermentation parameters (P > 0.05). The effects of A. horrida tannins may be strongly linked to their structure and their molecular weight more than to their concentration. For this reason, it is interesting that this work may be completed by physical characterization of these tannins.