We conducted an on-site application study at the livestock cooperative fertilizer plant to compare the composting period, temperature change, moisture content, and chemical properties between livestock manure compost using sawdust as a moisture regulator with those using spent oyster mushroom substrate. The composting period, moisture content, and fertilizer composition of compost containing spent oyster mushroom substrate did not differ from that of conventional compost mixed with sawdust after the first and second fermentation and post-maturation stages, it was suitable as a material for manufacturing livestock manure compost. The spent oyster mushroom substrate also lower the production cost of livestock manure compost by replacing the more expensive sawdust. The developed technology is expected to contribute towards the utilization of by-products of the oyster mushroom harvest while simultaneously producing high quality livestock manure compost.