Abstract Background The abuse of antibiotics in animal husbandry imposes a serious threat to both animal health and the environment. As a replacement for antibiotics, probiotic products have been widely used in livestock farming to promote growth of animals. However, no products specifically developed for farmed raccoon dogs and foxes are commercially available at the moment. This study was conducted to investigate the effects of mixed probiotics on farmed raccoon dogs and foxes. Results Two feeding trials on farmed raccoon dogs and foxes were performed. A mixed probiotic preparation composed of Bifidobacterium bifidum, Clostridium butyricum, Bacillus subtilis and Bacillus licheniformis was fed to these two canine species in order to assess whether such a mixed probiotics can be an alternative to antibiotics (control group). The body weight of raccoon dogs exhibited an increasing tendency with mixed probiotics administration, while that of foxes did not. The serum antioxidant activity was evaluated, and a significantly increase of total antioxidative capacity (T-AOC) was observed in both species. Illumina MiSeq was used for the sequencing of 16S rRNA genes to compare the composition of fecal microbiota between the control and mixed probiotics groups. Although α-diversity did not change, β-diversity of the fecal microbiota showed a distinct dissimilarity between the control and probiotics groups of both raccoon dogs and foxes. Dietary mixed probiotics increased the abundance of the genus Bifidobacterium in the fecal samples of raccoon dogs, and the genus Bacillus in the fecal samples of foxes. The different responses of raccoon dogs and foxes to probiotics might be the result of differences in the composition of the native gut microbiota of the two species. Conclusions The mixed probiotics preparation composed of Bifidobacterium bifidum, Clostridium butyricum, Bacillus subtilis and Bacillus licheniformis could be an effective feed additive for the improvement of the health of farmed raccoon dogs, but it may not be suitable for foxes.