<b><i>In Vitro</i></b><b> Digestion, Cytotoxicity, Microbial Viability of Vegan Soups, Human Intestinal Cell Models</b>

The viability of the Jarro Dophilus inoculum was determined by aseptically suspending the capsule content in saline solution. Bacterial viability was measured at the starting point (0 h) and after 24 hours of anaerobic incubation at 37 °C. MRS medium was used to culture Lactobacillus-related genera, while Garche medium (as described by Jarocki et al., 2014) was applied for Bifidobacterium strains. Viability was assessed through standard plate counting. To monitor bacterial survival in the “large intestine” phase of digestion, the inoculum was introduced, and samples were collected in triplicate both at time zero and after 24 hours of simulated digestion.This study evaluated the effects of in vitro digestion of functional vegan soups, rich in cholinesterase inhibitors, on human intestinal cells and gut bacteria. The research was conducted using normal colon epithelial cells (CCD 841 CoTr) and colorectal cancer cells (HT-29). Cytotoxicity was assessed using MTT and Neutral Red assays, while cell cycle distribution was analyzed via flow cytometry. Apoptosis and necrosis were evaluated with Annexin V/PI staining, and morphological changes were visualized using May-Grünwald-Giemsa staining. Bacterial viability was assessed through culture-based methods, and scanning electron microscopy (SEM) was used to examine bacterial adherence and matrix structure after digestion. The study provided insights into how digestion modifies the bioactivity of functional soups, their selective effects on normal and cancerous intestinal cells, and their impact on bacterial viability during the large intestine phase.