Objective: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection., Methods: From forty 4-5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR., Results: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm ([1.03 ± 0.09] vs [0.07 ± 0.03] %, P<0.01), grade b sperm ([2.07 ± 0.52] vs [0.35 ± 0.13] %, P<0.01), straight line velocity (VSL) ([10.95 ± 0.98] vs [6.78 ± 1.05] μm/s, P<0.01), curvilinear velocity (VCL) ([42.03 ± 1.35] vs [38.10 ± 7.65] μm/s, P>0.05), average path velocity (VAP) ([16.22 ± 1.52] vs [10.05 ± 1.80] μm/s, P<0.01), and the mRNA expressions of COX Ⅰ ([2.25 ± 0.24] vs [0.93 ± 0.10] %, P<0.01) and Ⅱ ([6.72 ± 0.37] vs [2.95 ± 0.78] %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: [1.11 ± 0.30] and [0.60 ± 0.19]%; grade b sperm: [2.40 ± 0.59] and [1.32 ± 0.27] %; VSL: [12.11 ± 1.62] and [11.47 ± 1.21] μm/s; VCL: [54.30 ± 2.35] and [45.75 ± 1.64] μm/s; VAP [18.40 ± 1.27] and [16.69 ± 1.02] μm/s; expression of COXⅠ mRNA: [1.86 ± 0.30] and [1.74 ± 0.17] %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01)., Conclusions: UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.