A trail was conducted to determine utilization of diet containing poultry excreta with different sources of nitrogen by cattle or buffalo rumen microorganisms in vitro. The aim was to find alternative sources of protein and low prices ruminant diets. Commercial broiler house excreta were used as 10% of a ration. Supplemented diets were basal diet with 10% poultry excreta alone or either with 3 g/kg yeast or 1.5 g urea/kg feed or both as well as 2% molasses. The basal diet comprised 40% concentrates and 60% wheat straw on dry matter (DM) basis as a control diet. All diets were inoculated with rumen fluid of cattle or buffalo. Digestibility of DM was not significantly influenced by both species and diets. However, organic matter (OM) digestion varied significantly with different diets, the highest value was obtained by yeast, urea, poultry excreta and molasses group (64.70%). Species and diets showed significant differences in neutral digested fiber (NDF) digestion. The highest values were obtained by buffalo and the yeast, urea, poultry excreta and molasses group (65.09%). Ammonia production was significantly increased by adding urea to the diet (12.64 and 13.79). Total volatile fatty acid and acetic acid % were produced significantly higher by cattle and the group of poultry excreta only. A buffalo rumen fluid produced higher propionic acid% and less acetic: propionic percent than cattle. The total number of microorganisms influenced significantly by diet only, as yeast, urea, poultry excreta and molasses group obtained the highest number.