Forty-eight rams, originating from a fauna-free flock, were divided into three groups of 16 and fed a corn silage-based diet that was unsupplemented (control) or included bentonite or monensin supplements. Eight rams in each group were inoculated with a mixed population of ruminal protozoa; the other rams remained free of protozoa throughout the 110-d experiment. The rams had free access to drinking water and assigned diet. All rams were killed at the end of the experiment, and ruminal and abomasal contents and livers were removed and sampled. Protozoal numbers in ruminal fluid of faunated rams were lower for groups fed bentonite or monensin supplements than for the control group. Bentonite decreased and monensin increased ruminal pH. The ruminal solubilities of Cu, Zn, and Mg were decreased by the presence of ruminal protozoa, but those of Fe, Mn, and Ca were not affected. Bentonite supplement decreased, and monensin supplement increased, the ruminal solubilities of Cu, Zn, and Mg. Protozoa increased the abomasal solubilities of Fe, but the other elements were not affected. Liver concentrations of Cu were decreased by bentonite and increased by monensin, but protozoa decreased the liver concentrations of both Cu and Mg. Liver concentration of Zn was affected by a monensin x protozoa interaction and that of Mg by a bentonite x protozoa interaction. It was concluded that chronic Cu poisoning could be accelerated by dietary supplements of monensin in sheep without ruminal microfauna, and the dietary Cu bioavailability could be decreased by dietary supplements of bentonite in sheep with a normal population of protozoa in the rumen.