The invention provides a preparation method of siberian tiger intestinal tract probiotics, which comprises the following steps that: first preparing the fermentation broth of five strains of probiotics, i.e. lactobacillus acidophilus, lactobacillus plantarum, lactococcus lactis, lactobacillus rhamnosus and bifidobacterium bifidum, removing supernatant at 5000r/min, respectively adopting 10g of mud of each type of probiotic, adding 4g of agaricus mushroom powder, mixing with 40g of mixed colloid containing 2.5 percent sodium alginate and 0.5 percent flaxseed gum, adding edible oil with 1.5 times of volume and 1.5 percent CaCl2 solution to be calcified for 3h, washing a gel ball for three times with saline water, putting the gel ball into 0.6 percent chitosan solution with the pH value of 5.5 to carry out film reaction for 15min and be taken out, flushing with the sterilizing saline water and putting into 55mM Citric acid solution to be liquefied for 30min to obtain the probiotic mud, finally cooling and drying to obtain intestinal tract probiotics powder, and obtaining the intestinal tract probiotics through the compounding of various probiotic powder.