[Objective] To explore the effect of adding compound probiotics on the growth performance, blood physiological and biochemical indexes and gut microbiota composition of beef cattle. [Methods] Eighteen crossbred 1 generation of Simmental and Holstein cattle were selected in Qinghai area, and randomly divided into two groups according to the principle of similar average weight of each group (n=9). The Y1 group was fed with compound probiotics containing Bacillus licheniformis, Bacillus subtilis and Saccharomyces cerevisiae.[Results] The daily increase in chest circumference was significantly increased in Y1 group (P>0.05); the concentration of cortisol was significantly higher in the Y1 group (P<0.05), and the concentration of high density liptein cholesterol, low density liptein cholesterol and total cholesterol was significantly lower in the Y1 group (P<0.05). Analysis the gut microbiota by 16S rRNA gene sequencing, there was not significant difference in Alpha diversity between two group (P>0.05), Beta diversity significantly differences of two group (P<0.05). The relative abundance of Tenericutes, Alisipes and Ruminococcaceae were significantly higher than that of the Y2 group (P<0.05), but the relative abundance of Alloprevotella was significantly lower than that of the Y2 group (P<0.05). There was not significant difference in the detected Bacillus sp.. Interestingly, the Y1 group reduced the abundance of E. coli in the gut. [Conclusion] Feeding compound probiotics can effectively reduce the synthesis of serum cholesterol and inhibit the reproduction of harmful microbial, which provides a theoretical basis for the healthy breeding of beef cattle.