Introduction: Gastrointestinal infections are one of the most important health challenges worldwide. One of the main causes of these infections is the bacterium Clostridium difficile, which can cause serious problems such as colitis, intestinal inflammation, and severe diarrhea. Common treatment methods include the use of antibiotics, which face problems such as bacterial resistance and side effects. Therefore, the need for new and effective strategies to deal with these infections is felt.

Purpose of the research: The purpose of this research is to investigate the effect of nanoemulsions containing vitamin C and D3 on the growth of Clostridium difficile bacteria. This research was conducted with the aim of investigating the ability to inhibit the growth of Clostridium difficile bacteria and improve intestinal health through the use of nanoemulsions containing vitamin C and D3.

Methodology: To investigate the effects of nanoemulsion on bacterial growth, mice were divided into three groups: the control group that received only bacterial suspension, the first group that received empty buttermilk with bacteria, and the second group that received buttermilk containing vitamin C and D3 nanoemulsion with bacteria. and the second group that received buttermilk containing vitamin C and D3 nanoemulsion along with bacteria. After 14 days, stool samples were collected from all groups and cultured on CCFA culture medium. Nanoemulsions were evaluated using different techniques including Dynamic Light Scattering (DLS) and zeta potential determination.

Results and discussion: The results showed that the nanoemulsion containing vitamin C and D3 has the ability to inhibit the growth of Clostridium difficile bacteria, so that the second group showed the lowest number of bacterial colonies. The histopathological analysis of the intestines of mice also showed that the vitamin nanoemulsion can cause significant changes in the tissue structure of the intestine. In the treatment group, a decrease in the density of the submucosal layers and a uniform distribution of goblet cells and lymphocytes in the mucosal layers were observed, which can show the improvement of the intestinal inflammatory condition and the reduction of inflammatory activity. These findings show that vitamin C and D3 nanoemulsion can act as an effective component in intestinal structural and inflammatory changes.

Conclusion: The results of this research suggest that nanoemulsions containing vitamin C and D3 can be used as a new and effective solution to control and prevent gastrointestinal infections and improve intestinal health. In addition, this research can lay the foundation for more research in the field of using nanoemulsions and natural compounds in the treatment of intestinal and infectious diseases.