Consecutive batch cultures (CBC), involving nine serial transfers at 3, 5 and 7 d intervals (21, 45 and 63 d, respectively) were established to enrich for plant fibre degrading co-cultures of anaerobic fungi and methanogens from rumen digesta. Microbial diversity and fermentation end-products were measured at appropriate intervals over each CBC time-course. While methanogenic populations remained diverse, anaerobic fungal diversity was related to transfer interval and appeared to decrease with increasing transfer number. Acetate was the principal aqueous fermentation end-product with minimal quantities of lactate and formate detected. Methane and carbon dioxide were detected in the gaseous head-space of all co-cultures and the total amounts of gas generated per transfer was greater with transfer intervals of 5 and 7 d compared with a 3 d interval, although the 3 d interval tended to be more efficient per unit time. In conclusion, rapidly growing, methane producing co-cultures of anaerobic fungi and methanogens from rumen digesta were easy to establish on lignocellulose (barley straw) and maintain over considerable time periods. These results suggest such co-cultures have potential in industrial scale anaerobic digestion (AD) of highly fibrous substrates, which are resistant to degradation in conventional AD plants.