The invention relates to a bio-padding and a preparation method thereof. The bio-padding comprises the following raw materials: saw dust, corn flour, soybean meal, crop straw, tapioca starch, potato starch, rice husk powder, wheat bran, bagasse, brown sugar, and biological flora. The bio-padding can completely decompose the excrement in the livestock and poultry breeding stage, so that in the breeding circle, no odor is generated, no mosquito flies grow, and no water is needed for washing; since no pollution is generated, the death rate of livestock and poultry is also reduced; and in addition, the fermentation waste produced by decomposing the excrement can be returned to the field as an organic fertilizer, so the recycling rate is high. The preparation method is simple and strong in controllability, and can realize large-scale production. The bio-padding solves the problems of arbitrary pollution and low utilization rate of the excrement during the livestock and poultry breeding process.