The annual forage legume biserrula (Biserrula pelecinus L.) offers a promising opportunity as a lowmethanogenic, bioactive pasture for southern Australian grazing systems where subterranean clover (Trifolium subterraneum L.) is the dominant annual pasture legume. This in vitro study to assess methanogenic potential examined how growth stage and cutting of biserrula affect biomass, nutritive value and fermentative parameters including methanogenic potential compared with subterranean clover. Both species were grown in a glasshouse, where three growth-stage treatments comprised plant collection at vegetative, reproductive or maturity growth stages. Three cutting (simulated grazing) treatments included cutting at the vegetative or reproductive stage and an uncut control, with herbage collection at maturity. Methane production in biserrula was unaffected by the stage of growth. Other nutritive value and fermentative parameters varied significantly with growth stage, the highest fibre content and lowest crude protein (CP) being found at maturity. Regrowth from herbage cut at the reproductive stage had higher CP and lower biomass than other cut treatments. In biserrula, this regrowth also showed the highest fermentability of the three cutting treatments including elevated methane yield. Notwithstanding these treatment effects on fermentability, biserrula maintained its strong antimethanogenic advantage over subterranean clover across all treatments, confirming its potential as an anti-methanogenic bioactive pasture.