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The addition of the inoculant can accelerate the fermentation process and improve the quality of the feed material in fermentation. This study aimed to isolate, test and select the rumen microorganisms that have high ability to degrade the fiber and improve the quality of materials used in inoculants to ferment agricultural wastes as alternative feed. Microbial isolate was extracted from cattle rumen fluid. The isolates type of fungus were grown on Potato Dextrose Agar (PDA) while the bacteria was grown on Nutrient Agar (NA) medium. Microbes of fungi were tested and selected based on their ability to produce ligninase, cellulase and hemicellulase enzymes. Bacteria were grown on MRS broth media as a confirmation for LAB colonies, then tested for gram staining, catalase test, growth on different NaCl levels (4 and 6.5%) and growth at various pH (3, 4, and 5). The results showed that five fungal isolates had a growth diameter of 0.1–0.37 mm on lignin, cellulose and hemicellulose media after seven days. Eleven bacterial isolates were indicated as lactic acid bacteria; almost all isolates could grow on NaCl 6.5%, but only four bacterial isolates could grow on NaCl 4% and five bacterial isolates could grow at pH 3. In conclusion, three isolates of fungi have a remarkable ability to degrade lignin. Five isolates of lactic acid bacteria have a comprehensive ability as a preservative in fermentation, so eight microbial isolates from rumen fluid can be used as inoculants in fermenting the feed material from agro-industrial waste.
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