ABSTRACTSalmonellais a foodborne pathogen that is one of the main causes of gastroenteric disease in humans and animals. As a natural organic substance, sodium humate (HNa) possesses antibacterial, antidiarrheal, and anti-inflammatory properties. However, it is unclear whether the HNa and HNa-derived microbiota exert alleviative effects onSalmonella entericaserovar Typhimurium-induced enteritis. We found that treatment with HNa disrupted the cell wall ofS. Typhimurium and decreased the virulence gene expression. Next, we explored the effect of HNa presupplementation onS. Typhimurium-induced murine enteritis. The results revealed that HNa ameliorated intestinal pathological damage. In addition, we observed that presupplementation with HNa enhanced intestinal barrier function via modulating gut microbiota, downregulating toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) and NOD-like receptor protein 3 (NLRP3) signaling pathways, regulating intestinal mucosal immunity, and enhancing tight junction protein expression. To further validate the effect of HNa-derived microbiota onS. Typhimurium-induced enteritis, we performed fecal microbiota transplantation and found that HNa-derived microbiota also alleviatedS. Typhimurium-induced intestinal damage. It is noteworthy that both HNa and HNa-derived microbiota improved the liver injury caused byS. Typhimurium infection. Collectively, this is the first study to confirm that HNa could alleviateS. Typhimurium-induced enteritis in a gut microbiota-dependent manner. This study provides a new perspective on HNa as a potential drug to prevent and treat salmonellosis.IMPORTANCESalmonellaTyphimurium is an important zoonotic pathogen, widely distributed in nature.S. Typhimurium is one of the leading causes of foodborne illnesses worldwide, and more than 350,000 people died fromSalmonellainfection each year, which poses a substantial risk to public health and causes a considerable economic loss. Here, we found that theS. Typhimurium infection caused severe intestinal and liver damage. In addition, we first found that sodium humate (HNa) and HNa-derived gut microbiota can alleviateS. Typhimurium infection-induced intestinal damage. These findings extend the knowledge about the public health risk and pathogenic mechanisms ofS. Typhimurium.