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This study screened six different species of forest plants and then further evaluated the most promising plant, giant milkweed (Calotropis gigantea), for the potential to improve nitrogen utilization efficiency (NUE) through inhibiting rumen protozoa in vitro. Ground leaves of giant milkweed at 1.6 and 3.2 mg/mL decreased the counts of Entodinium cells by 41.30% and 58.89%, respectively, and damaged their cell surface structure. Dasytricha, Isotricha, Epidinium, Ophryoscolex, and Diplodinium were not affected, while total bacterial and

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