The sub-acute toxicity of E. faecalis HZNU P2 was investigated in rats fed with different doses for 14 days. To evaluate the acute oral toxicity of E. faecalis HZNU P2, rats were fed with E. faecalis HZNU P2 at a high dose of 2×1011 CFU kg−1 for 10 days. Results showed that there were no abnormal clinical signs in any of the groups during the experiment. There were no significant differences in live weight gain among rats fed with E. faecalis HZNU P2, compared to those in control group. Macroscopic or microscopic examinations of organs revealed no abnormalities, indicating that E. faecalis HZNU P2 did not adversely affect the health of rats. Results of this study demonstrated that digestion of E. faecalis HZNU P2 in rats did not show any obvious signs of toxicity.