Objective To establish a sterile golden hamster diabetes model by fecal microbiota transplantation and evaluate the model. Method Twenty-four male sterile golden hamsters were divided into the control group (CN) and the model group (DM), with 12 hamsters in each group. They were respectively inoculated with fecal suspensions of healthy SD rats and diabetic SD rats. 10 weeks after fecal microbiota transplantation, the animals were euthanized. Blood, contents of the small intestine, pancreas and large intestine were collected for blood biochemistry, histopathological examination and 16S rRNA sequencing. Results After fecal microbiota transplantation, the detection of blood biochemical indicators showed that there were significant differences in apolipoprotein A1 and apolipoprotein B between the CN group and the DM group (P<0.05). In the DM model group, the levels of glucagon and fasting blood glucose were significantly increased (P<0.01), and fasting insulin was significantly decreased (P<0.001), presenting obvious characteristics of glycolipid metabolism disorder and insulin resistance. HE staining showed that compared with the CN group, the pancreas in the DM group presented with multifocal fat deposition, islet atrophy, reduction of islet cells, irregular islet shape, swelling and necrosis, islet cell proliferation, increased capillaries, and thickening of the basement membrane. In the DM group, there was hyperplasia of the small intestinal villus epithelium, fusion and disordered arrangement of the small intestinal villi, cell swelling accompanied by degeneration and necrosis, and aggregation of inflammatory cells in the lamina lining of the small intestinal mucosa. 16S rRNA sequencing showed that there were significant differences in the diversity of intestinal microbiota and the abundance of specific microbiota between the CN group and the DM group. The chao1 index, shannon index and simpson index in the α diversity index of the DM group were significantly decreased (P<0.05). The Unweighted Unifrac distance decreased significantly (P<0.01), suggesting that there were differences in β diversity. There were significant differences in the composition of intestinal bacterial communities between the two groups. In the DM group, the Verrucomicrobiota was significantly increased (P<0.05), and the Firmicutes was significantly decreased (P<0.05). Conclusion The sterile golden hamster diabetes model was successfully established through fecal microbiota transplantation technology. The animals presented clinical characteristics similar to those of diabetes, providing an animal model for in-depth research on the relationship between intestinal microbiota and the occurrence and development of diabetes.