| Summary: | <i>Salmonella</i> spp. remains a major public health problem for the whole world. Intestinal epithelial cells serve as an essential component of the mucosal innate immune system to defend against <i>Salmonella</i> infection. Our in vitro studies showed probiotics and active vitamin D have similar effects on innate immunity in <i>Salmonella</i>-infected intestinal epithelial cells, including antimicrobial peptide and inflammatory responses, to protect the host against infection while downregulating detrimental overwhelming inflammation. Hence, we investigated the synergistic effects of probiotics and active vitamin D on <i>Salmonella colitis</i> and translocation to liver and spleen by in vitro and in vivo studies. The <i>Salmonella</i> colitis model is conducted with 6–8 <i>w</i>/<i>o</i> male C57BL/6 mice: Streptomycin (20 mg/mouse p.o.)-pretreated C57BL/6 mice are mock infected with sterile PBS or infected orally with 1 × 10<sup>8</sup> CFU of a S. Typhimurium wild-type strain SL1344 for 48 h. The mice in the treated groups received 1, 25D daily (0.2 ug/25 g/d) and/or 1 × 10<sup>8</sup> CFU of probiotics, <i>Lactobacillus rhamnosus GG</i> (LGG) and <i>Bifidobacterium longum</i> (BL) by intragastric administration for 14 days. The in vivo study demonstrated the combination of probiotic <i>Bifidobacterium longum</i> and active vitamin D3 had the synergistic effects on reducing the severity of <i>Salmonella</i> colitis and body weight loss in C57BL/6 mice by reducing cecal inflammatory mIL-6, mIL-8, mTNF-α and mIL-1β mRNA responses, blocking the translocation of bacteria while enhancing the antimicrobial peptide mhBD-3 mRNA in comparison to the infection only group. However, LGG did not have the same synergistic effects. It suggests the synergistic effects of <i>Bifidobacterium longum</i> and active vitamin D on the antibacterial and anti-inflammatory responses in <i>Salmonella</i> colitis. Therefore, our in vivo studies demonstrated that the combination of probiotic <i>Bifidobacterium longum</i> and active vitamin D3 has the synergistic effects on reducing the severity of <i>Salmonella</i> colitis via the suppression of inflammatory responses, and blocking the translocation of bacteria through the enhancement of antimicrobial peptides.
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