This study evaluated the safety of Lactobacillus johnsonii IDCC 9203 and investigated its anti-inflammatory activity in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Genomic analysis revealed that this strain has no virulence and antibiotic resistance gene except tetW, which is a tetracycline resistance gene. Minimum inhibitory concentration data showed that the strain is resistant to tetracycline and aminoglycosides. Further analysis indicated that the transferability of the tetW gene is extremely low, and resistance to aminoglycosides is due to the intrinsic resistance of L. johnsonii IDCC 9203. Phenotypic safety assessment showed that the strain has neither β-hemolytic nor β-glucuronidase activity, and no biogenic amine production. When LPS-induced RAW 264.7 cells were treated with L. johnsonii IDCC 9203, the level of nitric oxide and expression of pro-inflammatory cytokines significantly decreased (p < 0.05). Therefore, L. johnsonii IDCC 9203 strain is considered as safe and beneficial probiotic for human consumption.

Objective: We investigated the anti-inflammatory and anti-pathogenic activities of Lacticaseibacillus rhamnosus IDCC 3201 isolated from the feces of breast-fed infants.Methods: Cell viability, nitric oxide (NO) production, and expression of inflammatory markers by L. rhamnosus IDCC 3201 were quantitatively analyzed in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. The antibacterial and antifungal activities of L. rhamnosus IDCC 3201 against various pathogens were also investigated.Results: Treatment of LPS-induced macrophages with cell-free supernatant of L. rhamnosus IDCC 3201 signifi-cantly decreased the expression levels of tumor necrosis factor (TNF-alpha) and interleukin-6 (IL-6). Nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) levels also significantly decreased in LPS-induced macrophages. Phenotypically, the treatment of L. rhamnosus IDCC 3201 reduced the production of nitric oxide (NO) in LPS-induced macrophages. Furthermore, L. rhamnosus IDCC 3201 was proven to have potent inhibitory activities against various pathogens responsible for inflammatory responses in the gastrointestinal tract (i.e., Bacillus ce-reus, Enterococcus faecalis, Staphylococcus aureus, and Salmonella Typhimurium), respiratory system (i.e., Strep-tococcus pneumoniae), and vagina (i.e., Candida albicans).Conclusion: L. rhamnosus IDCC 3201 has anti-inflammatory activity in terms of decreased expression of cytokines, inflammation-inducible enzymes in LPS-induced macrophages, and anti-pathogenic activity.

Abstract This study investigated the anti-inflammatory activity of L. plantarum IDCC 3501 isolated from kimchi (Korean fermented food) and its safety. When lipopolysaccharide (LPS)-induced RAW 264.7 macrophages were treated with cell-free supernatant from L. plantarum IDCC 3501, the mRNA expression level of inflammatory markers (i.e., TNF-α, IL-1β, and IL-6) was significantly reduced. The decreased cell viability by LPS was recovered and NO production in LPS-induced cell was also decreased. The genes responsible for antibiotic resistance and virulence were not detected from the genome analysis of this strain. Consistent with this, minimal inhibitory concentrations against various antibiotics, biogenic amines and D-lactate production, as well as enzymatic and hemolysis activities indicated that L. plantarum IDCC 3501 did not produce any harmful compounds during fermentation. Also, no acute toxicity and mortality were observed in a murine mouse model when feeding with L. plantarum IDCC 3501. Based on our findings, L. plantarum IDCC 3501 is safe and beneficial for human consumption.

Purpose Lactobacillus reuteri is one of the most largely studied human-commensal bacteria and widely used as a form of probiotics. Safety of probiotics has become increasingly important for human consumption due to increasing health-concerns in food industry. In this study, the safety of L. reuteri IDCC 3701 isolated from human breast milk was thoroughly investigated. Methods Whole-genome sequence analysis was performed to identify antibiotic resistance and toxigenicity of L. reuteri IDCC 3701. Phenotypic analysis such as minimal inhibitory concentration, β-hemolysis, extracellular enzyme activity, and the production of biological amines and L/D-lactate, was investigated. Finally, acute oral toxicity test was performed to access L. reuteri IDCC 3701 safety. Results Although multiple resistances to gentamicin and kanamycin were observed in L. reuteri IDCC 3701, it was revealed that these resistances are intrinsic and not transferable through whole-genome analysis. In addition, various phenotypic analysis concerning hemolysis, enzyme activity, and D-lactate production did not show any negative results. Although L. reuteri IDCC 3701 harbors a histidine decarboxylase gene, no biogenic amines were detected. Finally, L. reuteri IDCC 3701 exhibited no evidence of acute toxicity according to an in vivo study. Conclusion Our findings demonstrate that L. reuteri IDCC 3701 is considered to be safe for human consumption as probiotics based on the in silico, in vitro and in vivo studies.

Won Yeong Bang,, Seung A Chae, O-Hyun Ban, Sangki Oh, Chanmi Park, Minjee Lee, Minhye Shin, Jungwoo Yang*, and Young Hoon Jung*. Microbiol. Biotechnol. Lett. 2021;49:39-44. https://doi.org/10.48022/mbl.2011.11004

This study investigated the anti-inflammatory activity of Lactiplantibacillus plantarum IDCC 3501 isolated from kimchi (Korean fermented food) and its safety. When lipopolysaccharide (LPS)-induced RAW 264.7 macrophages were treated with cell-free supernatant from L. plantarum IDCC 3501, the mRNA expression level of inflammatory markers (i.e., TNF-α, IL-1β, and IL-6) was significantly reduced. In addition, the decreased cell viability by LPS was recovered and NO production in LPS-induced cell was also decreased. For the safety assessment, the genes responsible for antibiotic resistance and virulence were not detected from the genome analysis of this strain. Consistent with this, minimal inhibitory concentrations against various antibiotics, biogenic amines, and d-lactate production, as well as enzymatic and hemolysis activities, indicated that L. plantarum IDCC 3501 did not produce any harmful compounds during fermentation. Furthermore, no acute toxicity and mortality were observed in a murine mouse model. Based on our findings, L. plantarum IDCC 3501 is safe and beneficial for human consumption.

Abstract Safety evaluation of probiotics has become increasingly important for human consumption in food industry. The aims of this study were to assess safety of Streptococcus thermophilus IDCC 2201 through in vitro and in vivo tests. In results, this strain was found to be negative for hemolytic and β‐glucuronidase activity. In addition, thermophilus IDCC 2201 was susceptible to nine antibiotics suggested by EFSA. In accordance with MIC tests, whole‐genome analysis indicated that S. thermophilus IDCC 2201 neither harbors antibiotic resistance nor toxigenic genes. Furthermore, none of the biogenic amines including tyramine and histamine was produced and negligible amounts of D‐lactate were produced by S. thermophilus IDCC 2201. Finally, it was confirmed that there was no mortality and toxicity throughout single‐dose oral toxicity tests in rats. Therefore, we report that S. thermophilus IDCC 2201 is considered to be safe for human consumption as probiotics.