Citrus hystrix is widely used by Indonesians as a traditional medicine for gastrointestinal diseases, including Salmonella spp. infection. We investigated the antibacterial activity of the ethanolic peel extract of C. hystrix against Salmonella typhimurium.The antibacterial activity was evaluated both in vitro and in vivo. The minimum inhibitory concentration (MIC) of the extract was determined at a concentration of 0.625% by agar dilution assay. Later, the in vivo antibacterial activity was examined by the administration of 16mg of the extract daily for three consecutive days in a mouse model infected with S. typhimurium.The bacterial loads of S. typhimurium in the ileum, liver, and spleen decreased after 24h of administration of the extract (p=0.00008, p=0.00084, and p=0.00003, respectively).The ethanolic peel extract of C. hystrix shows antibacterial activity against S. typhimurium, indicating the potential of C. hystrix as an effective treatment for Salmonella spp. infection.