Lactic acid bacteria (LAB) form a group of bacteria to which most probiotics belong and are commonly found in fermented dairy products. Fermented foods and beverages are foods made through desired microbial growth and enzymatic conversions of food components. In this study, 43 LAB were isolated from Ethiopian traditional cottage cheese, cheese, and yogurt and evaluated for their functional and safety properties as candidate probiotics. Twenty-seven isolates, representative of each fermented food type, were selected and identified to the species level. Limosilactobacillus fermentum was found to be the predominant species in all samples studied (70.4%), while 11.1% of isolates were identified as Lactiplantibacillus plantarum. All 27 isolates tested showed resistance to 0.5% bile salt, while 26 strains were resistant to pH 3. The LAB isolates were also evaluated for antagonistic properties against key pathogens, with strain-specific features observed for their antimicrobial activity. Five strains from cottage cheese (Lactiplantibacillus plantarum 54B, 54C, and 55A, Lactiplantibacillus pentosus 55B, and Pediococcus pentosaceus 95E) showed inhibitory activity against indicator pathogens that are key causes of gastrointestinal infections in Ethiopia, i.e., Escherichia coli, Salmonella enterica subsp. enterica var. Typhimurium, Staphylococcus aureus, Shigella flexneri, and Listeria monocytogenes. Strain-specific immunomodulatory activity monitored as nuclear factor kappa B (NF-κB) and interferon regulatory factor (IRF) activation was documented for Lactiplantibacillus plantarum 54B, 55A and P. pentosaceus 95E. Antibiotic susceptibility testing confirmed that all LAB isolates were safe concerning their antibiotic resistance profiles. Five isolates (especially Lactiplantibacillus plantarum 54B, 54C, and 55A, Lactiplantibacillus pentosus 55B, and P. pentosaceus 95E) showed promising results in all assays and are novel probiotic candidates of interest for clinical trial follow-up.

Background: Lactiplantibacillus plantarum is a species found in a wide range of ecological niches, including vegetables and dairy products, and it may occur naturally in the human gastrointestinal tract. The precise mechanisms underlying the beneficial properties of these microbes to their host remain obscure. Although Lactic acid bacteria are generally regarded as safe, there are rare cases of the emergence of infections and antibiotic resistance by certain probiotics. Objective: An in silico whole genome sequence analysis of putative probiotic bacteria was set up to identify strains, predict desirable functional properties, and identify potentially detrimental antibiotic resistance and virulence genes. Methods: We characterized the genomes of three L. plantarum strains (54B, 54C, and 55A) isolated from Ethiopian traditional cottage cheese. Whole-genome sequencing was performed using Illumina MiSeq sequencing. The completeness and quality of the genome of L. plantarum strains were assessed through CheckM. Results: Analyses results showed that L. plantarum 54B and 54C are closely related but different strains. The genomes studied did not harbor resistance and virulence factors. They had five classes of carbohydrate-active enzymes with several important functions. Cyclic lactone autoinducer, terpenes, Type III polyketide synthases, ribosomally synthesized and post-translationally modified peptides-like gene clusters, sactipeptides, and all genes required for riboflavin biosynthesis were identified, evidencing their promising probiotic properties. Six bacteriocin-like structures encoding genes were found in the genome of L. plantarum 55A. Conclusions: The lack of resistome and virulome and their previous functional capabilities suggest the potential applicability of these strains in food industries as bio-preservatives and in the prevention and/or treatment of infectious diseases. The results also provide insights into the probiotic potential and safety of these three strains and indicate avenues for further mechanistic studies using these isolates.

Despite the widespread belief that “natural” traditional remedies are inherently safe, limited scientific evaluation raises important safety co