Chul Sung Huh

Breast milk-derived lactic acid bacteria (LAB) are promising sources of functional postbiotics. In this study, we evaluated the immunomodulatory, epithelial barrier-enhancing, and anti-melanogenic properties of nonviable HUH02 preparations, including heat-killed cells (HUH02-HK) and microfluidized preparations generated at 20,000 psi (HUH02-2psi) and 40,000 psi (HUH02-4psi), derived from the breast milk-derived Lactobacillus paragasseri HUH02 (L. paragasseri HUH02). HUH02 derivatives were evaluated in RAW264.7 macrophages, HaCaT keratinocytes, and B16F1 melanoma cells. All exhibited low cytotoxicity at functional concentrations. In Lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages, HUH02 derivatives markedly reduced nitric oxide production and suppressed the expression of inflammatory mediators, restored macrophage morphology toward a non-activated phenotype, and reduced iNOS protein expression. In HaCaT keratinocytes, HUH02 derivatives enhanced wound closure and the expression of epithelial barrier-associated genes, including ZO-1 and Occludin. Moreover, microfluidized derivatives, particularly HUH02-2psi and HUH02-4psi, significantly enhanced transepithelial electrical resistance, indicating improved epithelial barrier integrity. In B16F1 melanoma cells, HUH02-derived preparations exhibited antimelanogenic effects by reducing intracellular melanin accumulation and suppressing the expression of melanogenesis-related genes and proteins, including Tyr, TRP-1, TRP-2, and MITF. HUH02 was identified as L. paragasseri based on 16S rRNA, average nucleotide identity, and whole-genome phylogenetic analyses. Genome-based safety assessment revealed the absence of antibiotic resistance genes, virulence factors, plasmid replicons, and pathogenicity-associated markers. These findings demonstrated that breast milk-derived L. paragasseri HUH02 postbiotics exhibit multifunctional biological activities, including immunomodulatory, epithelial barrier-enhancing, and antimelanogenic effects.

Journal of Animal Science and Technology (JAST) is a peer-reviewed, open access journal publishing original research, review articles and notes in all fields of animal science. Topics covered by the journal include: genetics and breeding, physiology, nutrition of monogastric animals, nutrition of ruminants, animal products (milk, meat, eggs and their by-products) and their processing, grasslands and roughages, livestock environment, animal biotechnology, animal behavior and welfare.

Skin aging is influenced by structural alterations, oxidative stress, inflammation, and microbiome changes, and a comprehensive approach to addressing these factors may be effective for mitigating skin aging. This study evaluates the multifaceted anti-aging effects of heat-killed (HK-HN910) and lysed (LS-HN910) forms of Lactobacillus paragasseri HN910. Protective effects on cell viability, cell permeability, nitric oxide (NO) production, and skin anti-aging gene expression for both HK-HN910 and LS-HN910 were observed. Both forms significantly enhanced tight junction (TJ) protein zonula occludens- 1 ( ZO- 1 ) and antioxidant enzyme glutathione peroxidase ( GPx) gene expression, while significantly downregulating that of senescence-associated secretory phenotype pro-inflammatory cytokines interleukin ( IL ) - 1α , IL- 1β , IL- 6 , IL- 8 , and tumor necrosis factor-alpha ( TNFα ). LS-HN910 showed significantly greater upregulation of ZO- 1 and GPx and greater downregulation of IL- 1β and TNFα expression compared to HK-HN910. Cell wall component D-alanine (D-Ala) was released in higher amounts in LS-HN910 than in HK-HN910 and demonstrated anti-aging effects. D-Ala upregulated gene expression of skin barrier ZO- 1 , claudin- 1 ( Cla- 1 ), occludin ( OCC ), filaggrin ( FLG ), and sphingomyelin phosphodiesterase 2 ( SMPD2 ) and antioxidant enzymes superoxide dismutase ( SOD ), catalase ( CAT ), and GPx , while downregulating IL- 1α , IL- 1β , IL- 6 , IL- 8 , and TNFα . LS-HN910 treatment clinically revealed improvements in anti-aging parameters, including transepidermal water loss, skin water contents, sebum levels, dermal density, eye wrinkle index, skin pH, brightness, and microbiota composition, with a significant increase in Rhodococcus abundance. These findings indicate that LS-HN910, containing released D-Ala, is a promising cosmeceutical for preventing skin aging by enhancing the skin barrier, promoting oxidative defense, modulating inflammatory responses, and influencing skin microbiota.

Food Science of Animal Resources (Food Sci. Anim. Resour.) is an international, peer-reviewed bimonthly journal publishing original research and reviews on basic and applied aspects of the use of animal resources including the meat, egg, dairy, and edible insect/worm products. Food Sci. Anim. Resour. is published and distributed by the Korean Society for Food Science of Animal Resources.

Inflammatory bowel disease (IBD) is characterized by disruptions in the gut microbiome. While most studies on gut dysbiosis in IBD rely on sequencing-based methods, we employed a streamlined culturomics approach to obtain a more comprehensive understanding of gut microbiota imbalance in patients with IBD that may not be captured by sequencing alone. A total of 367 bacteria were identified at the species level, including 211 species from ulcerative colitis patients, 164 species from Crohn’s disease (CD) patients, and 263 species from healthy individuals. Consistent with our 16S rRNA gene amplicon sequencing results, a significant decrease in microbial diversity and a severe imbalance, especially in CD patients, were also observed in the culture-based analysis. Our culturomics approach provided additional insights, highlighting dysbiosis in unique anaerobic and Gram-negative species in CD patients. Moreover, species-level findings for Bifidobacterium and Enterobacterales emphasized specific species expansions in IBD patients. Notably, Mediterraneibacter gnavus, Thomasclavelia ramosa, Parabacteroides merdae, and Collinsella aerofaciens are of particular clinical interest due to their correlation with inflammatory biomarkers. This comprehensive analysis underscores the value of integrating a culture-based approach with a genome-based approach to provide complementary insights and therapeutic targets in IBD.

Bacterial culturomics is a set of techniques to isolate and identify live bacteria from complex microbial ecosystems. Despite its potential to revolutionize microbiome research, bacterial culturomics has significant challenges when applied to human gut microbiome studies due to its labor-intensive nature. Therefore, we established a streamlined culturomics approach with minimal culture conditions for stool sample preincubation. We evaluated the suitability of non-selective medium candidates for maintaining microbial diversity during a 30-day incubation period based on 16S rRNA gene amplicon analysis. Subsequently, we applied four culture conditions (two preincubation media under an aerobic/anaerobic atmosphere) to isolate gut bacteria on a large scale from eight stool samples of healthy humans. We identified 8141 isolates, classified into 263 bacterial species, including 12 novel species candidates. Our analysis of cultivation efficiency revealed that seven days of aerobic and ten days of anaerobic incubation captured approximately 91% and 95% of the identified species within each condition, respectively, with a synergistic effect confirmed when selected preincubation media were combined. Moreover, our culturomics findings expanded the coverage of gut microbial diversity compared to 16S rRNA gene amplicon sequencing results. In conclusion, this study demonstrated the potential of a streamlined culturomics approach for the efficient isolation of gut bacteria from human stool samples. This approach might pave the way for the broader adoption of culturomics in human gut microbiome studies, ultimately leading to a more comprehensive understanding of this complex microbial ecosystem.

The gut microbiota plays a pivotal role in metabolic disorders, notably type 2 diabetes mellitus (T2DM). In this study, we investigated the synergistic potential of combining the effects of Bifidobacterium longum NBM7-1 (CKD1) with anti-diabetic medicines, Lobeglitazone^Ⓡ (LO), Sitagliptin^Ⓡ (SI), and Metformin^Ⓡ (Met), to alleviate hyperglycemia in a diabetic mouse model. CKD1 effectively mitigated insulin resistance, hepatic steatosis, and enhanced pancreatic β-cell function, as well as fortifying gut-tight junction integrity. In the same way, SI-CKD1 and Met- CKD1 synergistically improved insulin sensitivity and prevented hepatic steatosis, as evidenced by the modulation of key genes associated with insulin signaling, β-oxidation, gluconeogenesis, adipogenesis, and inflammation by qRT-PCR. The comprehensive impact on modulating gut microbiota composition was observed, particularly when combined with Metformin^Ⓡ. This combination induced an increase in the abundance of Rikenellaceae and Alistipes related negatively to the T2DM incidence while reducing the causative species of Cryptosporangium, Staphylococcaceae, and Muribaculaceae. These alterations intervene in gut microbiota metabolites to modulate the level of butyrate, indole-3-acetic acid, propionate, and inflammatory cytokines and to activate the IL-22 pathway. However, it is meaningful that the combination of B. longum NBM7-1(CKD1) reduced the medicines' dose to the level of the maximal inhibitory concentrations (IC50). This study advances our understanding of the intricate relationship between gut microbiota and metabolic disorders. We expect this study to contribute to developing a prospective therapeutic strategy modulating the gut microbiota.

Type 2 diabetes (T2D) is known as adult-onset diabetes, but recently, T2D has increased in the number of younger people, becoming a major clinical burden in human society. The objective of this study was to determine the effects of Bifidobacterium and Lactiplantibacillus strains derived from the feces of 20 healthy humans on T2D development and to understand the mechanism underlying any positive effects of probiotics. We found that Bifidobacterium longum NBM7-1 (Chong Kun Dang strain 1; CKD1) and Lactiplantibacillus rhamnosus NBM17-4 (Chong Kun Dang strain 2; CKD2) isolated from the feces of healthy Korean adults (n = 20) have anti-diabetic effects based on the insulin sensitivity. During the oral gavage for 8 weeks, T2D mice were supplemented with anti-diabetic drugs (1.0–10 mg/kg body weight) to four positive and negative control groups or four probiotics (200 uL; 1 × 10⁹ CFU/mL) to groups separately or combined to the four treatment groups (n = 6 per group). While acknowledging the relatively small sample size, this study provides valuable insights into the potential benefits of B. longum NBM7-1 and L. rhamnosus NBM17-4 in mitigating T2D development. The animal gene expression was assessed using a qRT-PCR, and metabolic parameters were assessed using an ELISA assay. We demonstrated that B. longum NBM7-1 in the CKD1 group and L. rhamnosus NBM17-4 in the CKD2 group alleviate T2D development through the upregulation of IL-22, which enhances insulin sensitivity and pancreatic functions while reducing liver steatosis. These findings suggest that B. longum NBM7-1 and L. rhamnosus NBM17-4 could be the candidate probiotics for the therapeutic treatments of T2D patients as well as the prevention of type 2 diabetes.