The objective of this study was to examine the effects of Tithonia diversifolia as a supplementary forage on dairy cow performance and methane production. Nine lactating Holstein × Zebu dairy cows (519 ± 53.3 kg of body weight and 66 ± 13.3 d in milk) were paired by milk yield (21.3 ± 2.34 kg/d) and body weight and randomly assigned to three dietary treatments in a Latin square design with 21-d experimental periods (14 d for diet adaptation and 7 d for measurements and sample collection). The dietary treatments included the control diet consisting of fresh sugar cane plus concentrate (44:56, % of diet DM), and two treatment diets containing different levels of fresh T. diversifolia (6.5 and 15.4%, DM basis) which partially replaced both sugarcane and concentrates. Methane production was measured using the sulphur hexafluoride (SF6) technique from d 16 to d 21 of each experimental period. Analysis of the gas samples was performed by gas chromatography. The inclusion of T. diversifolia at 15.4% DM had no effects on DM intake, milk production, nitrogen balance or methane production. There was no effect on the concentrations of total saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) in milk fat (P ≥ 0.28), though individual milk fatty acids were affected. Serum concentrations of glucose, urea nitrogen (BUN), triglycerides, β-hydroxybutyrate (BHBA), and cholesterol were unaffected by the dietary treatments (P ≥ 0.13). There was a time (2 and 6 h post-feeding) and dietary treatment effect (P < 0.01) on the acetate to propionate ratio in the rumen. A denaturing gradient gel electrophoresis analysis of the archaeal community showed distinct clustering of the archaea populations for control and treatment diets. Taken together, our results indicate the potential of T. diversifolia as a supplementary forage for dairy cattle in the tropics.