Abstract: In order to ensure the comparability and reliability of animal test results, the stability of quality of tested animal must be kept, so it is very necessary to detect the quality of tested animals. One mouse was taken from 13 separation facilities of Compare Medical Center of Yangzhou University separately, and contents of ileocecal junction and life waste were collected and inoculated into responsible mediums. According to GB/T 14926.14 and GB/T 14926.17-2001, Staphylococcus aureus and Pseudomonas aeruginosa were detected. Results showed that the Staphylococcus aureus was positive in some mice, while Pseudomonas aeruginosa was negative in all mice. It is suggested that a part of mice in separation facilities of Compare Medical Center of Yangzhou University took Staphylococcus aureus, but there was no Pseudomonas aeruginosa.