This study consisted of two experiments with the following objectives: to evaluate the effects of tannins from the tropical legume macrotiloma (Macrotyloma axillare) on total gas and methane (CH₄ ) production, as well as on ruminal fermentation parameters by performing an in vitro bioassay, with samples incubated with and without polyethylene glycol (PEG) in a semi-automatic system; and secondly in a 17 day in vivo experiment, to determine apparent total tract digestibility (ATTD) of dietary nutrients and ruminal fermentation parameters of 12 intact 8- to 9-month-old Santa Inês (averaging 24.95 ± 1.8 kg body weight) ewes fed tropical grass hay supplemented with macrotiloma hay. The ewes were divided into two treatment groups depending on their diet: chopped aruana grass hay (Panicum maximum cv. Aruana) (control-CON); and aruana grass hay supplemented with chopped macrotiloma hay (macrotiloma-MAC). The animals were kept for 5 consecutive days in metabolic cages for the ATTD assay, and at the end of this period, samples of rumen fluid were collected from each ewe to determine ammoniacal nitrogen (NH₃ -N) and short-chain fatty acid (SCFA) production, and protozoa count. For the in vitro assay, a decrease in total gas and CH₄ production was observed for samples incubated without PEG (p < .05). No differences were observed for the other parameters evaluated (p > .05). In the in vivo experiment, increased intake and ATTD of crude protein were observed for the animals fed MAC when compared to CON (p < .05). For rumen fermentation parameters, increased NH₃ -N, total SCFA and isobutyrate concentrations, as well as reduced protozoa count were observed for MAC when compared to CON (p < .05). The results observed here indicated the potential of macrotiloma for use as a ruminant feed, and antimethanogenic potential of this plant was noted.

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