The main objective of this study was to compare the effect of supplementing beef cattle with Desmanthus virgatus cv. JCU2, D. bicornutus cv. JCU4, D. leptophyllus cv. JCU7 and lucerne on in vivo methane (CH4) emissions measured by open-circuit respiration chambers (OC) or the GreenFeed emission monitoring (GEM) system. Experiment 1 employed OC and utilized sixteen yearling Brangus steers fed a basal diet of Rhodes grass (Chloris gayana) hay in four treatments—the three Desmanthus cultivars and lucerne (Medicago sativa) at 30% dry matter intake (DMI). Polyethylene glycol (PEG) was added to the diets to neutralize tannin binding and explore the effect on CH4 emissions. Experiment 2 employed GEM and utilized forty-eight animals allocated to four treatments including a basal diet of Rhodes grass hay plus the three Desmanthus cultivars in equal proportions at 0%, 15%, 30% and 45% DMI. Lucerne was added to equilibrate crude protein content in all treatments. Experiment 1 showed no difference in CH4 emissions between the Desmanthus cultivars, between Desmanthus and lucerne or between Desmanthus and the basal diet. Experiment 2 showed an increase in CH4 emissions in the three levels containing Desmanthus. It is concluded that on high-quality diets, Desmanthus does not reduce CH4 emissions.