Buffalo milk contain high fat and protein, it is easy to cause the texture such as hard texture, poor fineness and low acidity. In this paper, single factor test and box-behnken center combination design were used to screen the fermentation and technological formulation of the fermented milk, and to test the physical and chemical and hygienic safety indicators. The results showed that when the rario of lactobacillus Bulgarian, thermophilic streptococcus and bacilli of the plant = 2:1:3, the bacterial species added amount of 4%, sugar added amount of 8%, fermentation time for 6h, the buffalo fermented milk was with rich nutrition value, fat content was 4.04%, protein content was 3.31%, lactobacillus content was 3.3×107CFU/mL, the index of microorganism was up to the standard. The study provides a reference basis for the production and processing of buffalo fermented milk.