Complete Genome Analysis of TS5 and Its Potential As a Probiotic Strain in Mice

Introduction: In recent years, a large number of studies have shown that has the potential as an animal feed additive, and its potential probiotic properties have been gradually explored.

Methods: In this study, Illumina NovaSeq PE150 and Oxford Nanopore ONT sequencing platforms were used to sequence the genome of TS5, a fiber-degrading strain isolated from Tibetan sheep. To further investigate the potential of TS5 as a probiotic strain, in vivo experiments were conducted using 40 five-week-old male specific pathogen-free C57BL/6J mice. The mice were randomly divided into four groups: high fiber diet control group (H group), high fiber diet probiotics group (HT group), low fiber diet control group (L group), and low fiber diet probiotics group (LT group). The H and HT groups were fed high-fiber diet (30%), while the L and LT groups were fed low-fiber diet (5%). The total bacteria amount in the vegetative forms of TS5 per mouse in the HT and LT groups was 1 × 10 CFU per day, mice in the H and L groups were given the same volume of sterile physiological saline daily by gavage, and the experiment period lasted for 8 weeks.

Results: The complete genome sequencing results of TS5 showed that it contained 3,929,788 nucleotides with a GC content of 46.50%. The strain encoded 3,873 genes that partially related to stress resistance, adhesion, and antioxidants, as well as the production of secondary metabolites, digestive enzymes, and other beneficial nutrients. The genes of this bacterium were mainly involved in carbohydrate metabolism, amino acid metabolism, vitamin and cofactor metabolism, biological process, and molecular function, as revealed by KEGG and GO databases. The results of mouse tests showed that TS5 could improve intestinal digestive enzyme activity, liver antioxidant capacity, small intestine morphology, and cecum microbiota structure in mice.

Conclusion: These findings confirmed the probiotic effects of TS5 isolated from Tibetan sheep feces and provided the theoretical basis for the clinical application and development of new feed additives.