The invention discloses a preparation method of a microbial feed based on a straw fermentation technology. The preparation method comprises the following steps: preparing a microbial leavening agent, wherein the microbial leavening agent comprises the following substances: saccharomycetes, bacillus, lactic acid bacteria, bifidobacterium and cellulase; according to the preparation method of the microbial feed based on the straw fermentation technology, the finished microbial feed contains rich functional substances such as viable bacteria, enzyme groups, vitamins, lipids, organic acids, amino acids, various oligosaccharides and coenzyme factors, small peptides and the like, few infectious microbes exist, and the feeding effect is obvious; the feed utilization rate is improved; the compound microbial agent can effectively improve the immunity and resistance of fed objects, replaces antibiotics, further improves the quality of animal products, eliminates stink of a pigsty, has obvious ecological benefits, can degrade hemicelluloses in the feed into monosaccharides or oligosaccharides after being fermented by using the compound microbial agent, relieves the hindering effect of the monosaccharides or oligosaccharides on nutritional factors, and improves the feed utilization rate. Furthermore, the nutrition resistance is eliminated, and the digestibility and the utilization rate of nutritional ingredients are improved.