This study investigated the effects of three pine-based biochar products on nutrient disappearance, total gas and methane (CH4) production, rumen fermentation, microbial protein synthesis, and rumen microbiota in a rumen simulation technique (RUSITEC) fed a barley-silage-based total mixed ration (TMR). Treatments consisted of 10 g TMR supplemented with no biochar (control) and three different biochars (CP016, CP024, and CP028) included at 20 g·kg?1 DM. Treatments were assigned to 16 fermenters (n = 4 per treatment) in two RUSITEC units in a randomized block design for a 17 d experimental period. Data were analyzed using MIXED procedure in SAS, with treatment and day of sampling as fixed effects and RUSITEC unit and fermenters as random effects. Biochar did not affect nutrient disappearance (P > 0.05), nor total gas or CH4, irrespective of unit of expression. The volatile fatty acid, NH3-N, total protozoa, and microbial protein synthesis were not affected by biochar inclusion (P > 0.05). Alpha and beta diversity and rumen microbiota families were not affected by biochar inclusion (P > 0.05). In conclusion, biochar did not reduce CH4 emissions nor affect nutrient disappearance, rumen fermentation, microbial protein synthesis, or rumen microbiota in the RUSITEC.