The objective of this study was to evaluate antimethanogenic activity of eucalyptus oil (EUC) and anise oil (ANI) in vitro and in vivo using sheep as a model. In vitro study was conducted using batch culture technique, each of EUC and ANI were added at 0, 50, 100, 200, or 400 mg/L of fermentation media with substrate containing 60% corn-based concentrate and 40% hay (DM basis). Total gas production (GP) linearly (P < 0.01) decreased with increasing ANI, whereas the GP was not affected with EUC addition. Supplementation of ANI and EUC linearly (P < 0.01) decreased total methane production and methane proportion in total gas. Total VFA and ammonia-nitrogen (NH3-N) concentration linearly (P < 0.01) decreased with increasing ANI supplementation. For the in vivo study, a replicated 3 × 3 Latin square design was carried out using six ruminal cannulated Du Han hybrid sheep (BW, 64.5 ± 8.56 kg) with 22 d of periods. Three treatments were control diet (consisted of 60% corn-based concentrate and 40% Chinese wildrye hay), EUC (control diet supplemented with 0.5 g EUC/d per head), and ANI (control diet supplemented with 0.5 g ANI/d per head). Each period consisted of 14 d for adaption and 8 d for sampling and data collection. Supplementation of EUC and ANI had no effects on feed intake and apparent nutrient digestibility. Ruminal NH3-N concentration was greater with EUC (P < 0.01) and ANI (P = 0.03) than control. Urinal allantoin output was less (P < 0.05) in sheep fed EUC and ANI than control animals. Methane emission was less (P = 0.03) in sheep fed ANI than sheep fed EUC, and a tendency of decrease for an eduction in this parameter was found for sheep fed with ANI (P = 0.08) compared to control. The in vitro results indicated a reduction of methane production with both EUC and ANI but in a dose-dependant manner. Supplementation of ANI tended to reduce ruminal methane production without adversely affecting rumen fermentation characteristics, nutrient intake, and digestibility, suggesting potential inhibition of ruminal methane emission in sheep supplemented with ANI.
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