Context Approximately 40–50% of all infertility cases are due to male infertility, and one of the most important causes of infertility is azoospermia. Aims This study aimed to evaluate the potential effect of elderberry on the spermatogenesis process in the azoospermia mice model. Method Thirty adult male mice were randomised into three groups: control; busulfan (45 mg/kg); and busulfan + elderberry (2%), 6 mL orally per animal. Sperm samples were collected from the tail of the epididymis, and testis specimens were also collected and then subjected to sperm parameters analysis, histopathological evaluation, reactive oxygen species (ROS), and glutathione (GSH) measurement to determine the mRNA expression and hormonal assay. Conclusions It can be concluded that the elderberry diet may be considered a complementary treatment to improve the spermatogenesis process in busulfan-induced azoospermic mice. Implications Considering some limitations, the elderberry diet can be an alternate option for improving testicular damage following chemotherapy.