This study was conducted to investigate the effects of sorbitol, mannitol, fructooligosaccharides (FOS) and raffinose on sperm quality, reproductive and immune organs of mice. 30 male mice were randomly divided into five groups, each group contained 6 mice (each mouse was used as a repeat). The mice of control group were fed a basal diet, and the mice of groups A, B, C and D were fed the experimental diets supplemented with five percent (w/w) of sorbitol, mannitol, FOS and raffinose, respectively. The experimental period was 30 days. The results showed as follows: sorbitol, mannitol, FOS or raffinose supplementation had no significant effects on sperm density and deformity rate (P>0.05). Compared with the control group, the sperm activity in group of sorbitol or raffinose was significantly decreased (P<0.05). The average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), lateral head displacement (ALH) and mobile of average degree (MAD) of sperm in experimental groups were lower than control group. Sorbitol supplementation significantly reduced the VAP, VCL and ALH of sperm (P<0.05). The FOS supplementation decreased the ALH of sperm (P<0.05). Raffinose supplementation significantly reduced VAP, VSL, VCL, ALH and MAD of sperm (P<0.05).The beat cross frequency (BCF) of sperm in group supplemented with mannitol or FOS was higher than control group, but not statistically significant (P >0.05). The linearity (LIN) and straightness (STR) of sperm in each group had no significant difference (P>0.05).There was no difference in organ index (testis, seminal vesicle, spleen and thymus) between each group (P>0.05). These finding showed that basal diet respectively supplemented with 5% sorbitol, 5% mannitol, 5%FOS or 5%raffinose had some negative effect on sperm quality in mice. But, not influence the development of reproductive and immune organs in mice.