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The mixed rumen microorganisms were obtained from a ruminally fistulated Hereford cow fed twice a day (06:00 and 16:00 h), a ration consisting of 70% corn silage, 20% alfalfa hay, 9% soybean meal, trace minerals, and vitamins (13% crude protein diet). Ruminal contents were collected from the bottom of the rumen 4 h after the morning feeding, squeezed through four layers of cheese cloth and poured into a separating funnel that had been gassed with oxygen-free CO 2 and anaerobically incubated at 39C for up to 60 min to allow feed particles to buoy up. The feed particles that had risen to the surface was removed by a vacuum tube, and then liquid portions were anaerobically filtered using nylon cloth (25 mm pore size) to remove small feed particles. The filtered liquid portions were used as a source of mixed rumen microorganisms.
Lee et al. (Wed,) studied this question.