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Dancers experience physical and psychological stressors that can impact gut health. We hypothesized that a three-month supplementation with Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 would result in measurable alterations in the fecal metabolomic profile and improve gastrointestinal symptomatology in dancers. Of the 51 volunteers, 26 female dancers were randomized to a 12-week trial (NCT05567653). A homogenous group of 16 (probiotic: n = 5; placebo: n = 11) was analyzed. The participants received L. helveticus R0052 and B. longum R0175 (3 × 109 colony-forming units/day) or a placebo. Baseline dietary intake and body composition were recorded. Fecal samples were analyzed using liquid chromatography–mass spectrometry, and gastrointestinal symptoms were assessed with the Rome IV questionnaire. Statistical methods included principal component analysis, mixed-effects models, and analysis of variance–simultaneous component analysis (ASCA). The study revealed shifts in the probiotic group’s fecal metabolome (permutation test p = 0.026), including a reduction in (2RS)-2-(4-hydroxyphenyl)propionic acid (p = 0.023). No improvement in gastrointestinal symptoms was observed. No adverse events occurred. L. helveticus R0052 and B. longum R0175 may alter the gut metabolome, notably (2RS)-2-(4-hydroxyphenyl)propionic acid, but small sample size and absent symptom improvement limit the conclusions. Larger studies with varied doses and blood metabolite analysis are needed to confirm relevance.

Background The burden of acute gastroenteritis on children and their families continues to be enormous. Probiotics, defined as viable microbial preparations that have a beneficial effect on the health of the host, represent a rapidly expanding field. Although clinical trials in children with gastroenteritis have been performed, most have significant flaws, and guidelines do not consistently endorse their use. Methods/Design PROGUT is a randomized, placebo-controlled, double-blind, five-center, Canadian, emergency department trial. Children aged 3 months to 48 months who present between November 2013 and June 2017 with <72 hours of gastroenteritis symptoms will be assessed for eligibility. A total of 886 children will be randomized (1:1 allocation via an internet based, third party, randomization service) to receive 5 days of a combination probiotic agent ( Lactobacillus rhamnosus and L. helveticus ) or placebo. All participants, caregivers, and outcome assessors will be blinded to group assignment. The study includes three key outcomes: 1) clinical - the development of moderate to severe disease following an emergency department (ED) evaluation that employs a validated clinical score (Modified Vesikari Scale); 2) safety - side effect; and 3) mechanism - fecal secretory immunoglobulin A levels. Discussion Definitive data are lacking to guide the clinical use of probiotics in children with acute gastroenteritis. Hence, probiotics are rarely prescribed by North American physicians. However, the following current trends obligate an urgent assessment: 1) probiotics are sold as food supplements, and manufacturers can encourage their use while their relevance has yet to be established; 2) North American and European government agencies remain concerned about their value and safety; 3) some institutions are now recommending the routine use of probiotics; and 4) parents of affected children are often providing probiotics. With probiotic consumption increasing in the absence of solid evidence, there is a need to conduct this definitive trial to overcome the limitations of prior work in this field. Trial registration ClinicalTrials.gov: NCT01853124 ; first registered 9 May 2013.

Lactobacillus helveticus is a probiotic bacterium widely used in the food industry. In this study, we evaluate the safety and probiotic properties of L . helveticus strain KM7, isolated from the gut of Apis cerana , through whole-genome sequencing and in vitro experiments. The complete genome consists of 2,164,024 bp, encoding 2192 genes with an average GC content of 36.82%. The strain lacks identifiable virulence factor genes and antibiotic resistance genes, shows no hemolytic activity, and cannot produce biogenic amines such as spermine, cadaverine, or putrescine. L. helveticus KM7 exhibited strong acid and bile salt tolerance, superior adhesion capacity, and remarkable antioxidant activity, with its genome harboring genes associated with these traits. It contains gene clusters potentially involved in the biosynthesis of bacteriocins Helveticin J and Enterolysin A, as well as encoding multiple genes of carbohydrate-active enzymes. Moreover, KM7 demonstrated excellent exopolysaccharide-producing capability, and glucose supplementation in the medium enhanced yield. These findings indicate that KM7 is a safe strain with promising probiotic properties, highlighting its potential for food applications.

Background and objective Lactic acid bacteria (LAB) are widely recognized for their probiotic and antimicrobial potential in food and health-related applications. This study aimed to isolate and characterize Lactobacillus helveticus strain CMI1 from traditional Iranian Khiki cheese and to evaluate its probiotic-related properties, including antimicrobial, anti-biofilm, antioxidant, and cytotoxic activities. Methods The isolate was identified by 16S rRNA gene sequencing. Antimicrobial activity of acidic and neutralized cell-free supernatants (CFS) was evaluated against major Gram-positive and Gram-negative foodborne pathogens using disk diffusion, well diffusion, modified double-layer, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. Anti-biofilm activity against Staphylococcus aureus was assessed using crystal violet staining and quantitative real-time PCR analysis of biofilm-related genes. Antioxidant activity was determined using DPPH, ABTS, and β-carotene/linoleic acid assays. Cytotoxic effects were evaluated against HT-29, HeLa, and MCF-7 cancer cell lines using the MTT assay. Antibiotic susceptibility was also examined. Results L. helveticus CMI1 exhibited strong antimicrobial activity, with higher efficacy observed for acidic CFS compared to neutralized CFS, particularly against S. aureus . The strain significantly inhibited S. aureus biofilm formation and downregulated key biofilm- and virulence-related genes ( mecA , agr , spa , icaA , and hla ). Additionally, CMI1 demonstrated notable antioxidant activity and dose-dependent cytotoxic effects against cancer cell lines, while remaining sensitive to clinically relevant antibiotics. Conclusion These findings indicate that L. helveticus CMI1 possesses promising probiotic and functional properties, supporting its potential application in food preservation and functional food development.

The liver is an important digestive gland, and acute liver failure results in high mortality. Probiotics are considered potential adjuvant therapies for liver disease. This study aimed to investigate the beneficial effects of Lactobacillus helveticus R0052 on acute liver injury and the underlying mechanisms. Sprague-Dawley rats were gavaged with L. helveticus R0052 suspensions (3 × 10 9 CFU) for 1 week. Subsequently, acute liver injury was induced by intraperitoneal d -galactosamine injection on the eighth day. After 24 h, samples (blood, liver, ileum, faeces) were collected and assessed for histological injury, inflammation, intestinal barrier, gut microbiome and metabolome. L. helveticus R0052 alleviated aminotransferase, bilirubin and total bile acid elevation and histological hepatic injuries. Additionally, L. helveticus R0052 exhibited anti-inflammatory properties by downregulating Toll-like receptors , tumour necrosis factor-α and nuclear factor-κb transcription in liver samples and decreasing proinflammatory cytokine plasma concentrations. Additionally, L. helveticus R0052 ameliorated intestinal abnormalities and regulated Toll-like receptors , claudin2 and mucin3 gene transcription in the intestine. These effects were associated with gut microbiome and metabolome modulation by L. helveticus R0052. Probiotic pretreatment enriched Lactobacillus and Bacteroides and depleted Flavonifractor and Acetatifactor in the gut microbiome. Meanwhile, L. helveticus R0052 improved carbohydrate and fatty acid metabolism and reduced lithocholic acid levels. These results indicate that L. helveticus R0052 is promising for alleviating acute liver injury and provide new insights regarding the correlations among the microbiome, the metabolome, the intestinal barrier and liver disease.

Probiotics are gaining attention for their benefits as a supplement to improve oral health. This study aimed to evaluate the antibacterial effect of the probiotic Lactobacillus helveticus against Porphyromonas gingivalis , a significant pathogen in periodontal diseases. Antibacterial susceptibility was assessed using the well diffusion assay, with 0.12% chlorhexidine (CHX) served as the positive control. Biofilm biomass was evaluated using crystal violet staining. Cell viability in P. gingivalis treated with L. helveticus was determined using the LIVE/DEAD Baclight bacterial assay via fluorescence microscopy. Ultra-morphological alterations in these cells were further examined using Field Emission Scanning Electron Microscopy. The results indicated that L. helveticus significantly reduced the growth of P. gingivalis . The highest concentration of 10 9 cells/mL achieved the most substantial inhibition in the well diffusion assay, followed by concentrations of 10 8 cells/mL and 10 7 cells/mL, which demonstrated a clear dose-dependent response. Furthermore, biofilms of P. gingivalis treated with L. helveticus exhibited a notable biomass reduction of up to 85% at the highest concentrations. LIVE/DEAD staining confirmed a decreased in cell viability among the treated populations, while FESEM analysis revealed morphological disruptions in P. gingivalis cells treated with L. helveticus . These findings suggest that L. helveticus has a potent antibacterial effect against P. gingivalis , highlighting the need for further research to identify the optimal probiotic strategies that could enhance periodontal health.

Traditional Iranian cheeses, especially “Khiki cheese,” represent valuable sources of indigenous lactic acid bacteria with potential probiotic properties. This study evaluated the probiotic attributes, safety profile, antimicrobial efficacy, antioxidant capacity, and anti-biofilm activity of Lactobacillus helveticus BGTRM7-58, a strain isolated from Khiki cheese. The strain demonstrated considerable anti-adhesion capability against Staphylococcus aureus . The cell-free supernatant (CFS) exhibited potent antimicrobial activity, displaying a minimum inhibitory concentration (MIC) of 15.625 mg/mL against S. aureus . Furthermore, the CFS inhibited biofilm formation by 87% at 4× MIC and disrupted pre-established mature biofilms by 89% at the same concentration. In vitro cytotoxicity assessment revealed dose-dependent antiproliferative effects against cancer cell lines. Quantitative real-time PCR analysis indicated significant downregulation of key staphylococcal virulence genes, most notably a 47% reduction in agr expression. The strain also exhibited substantial antioxidant activity, scavenging 68.67%, 71.75% of DPPH and ABTS radicals, respectively. Comprehensive safety evaluation confirmed the absence of hemolytic and DNase activities, no production of biogenic amines, and susceptibility to clinically relevant antibiotics. Taken together, these findings indicate that L. helveticus BGTRM7-58 fulfills the fundamental criteria for a safe and functional probiotic strain, highlighting its potential for application in functional food formulations and strategies aimed at controlling biofilm-associated infections.

ABSTRACT For the present study, we collected 22 Lactobacillus helveticus strains from different dairy (n = 10) and cereal (n  = 12) fermentations to investigate their biodiversity and to uncover habitat-specific traits. Biodiversity was assessed by comparison of genetic fingerprints, low-molecular-weight subproteomes, metabolic and enzymatic activities, growth characteristics and acidification kinetics in food matrices. A clear distinction between the dairy and cereal strains was observed in almost all examined features suggesting that the different habitats are domiciled by different L. helveticus biotypes that are adapted to the specific environmental conditions. Analysis of the low-molecular-weight subproteome divided the cereal isolates into two clusters, while the dairy isolates formed a separate homogeneous cluster. Differences regarding carbohydrate utilization were observed for lactose, galactose, sucrose and cellobiose as well as for plant-derived glucosides. Enzymatic differences were observed mainly for ß-galactosidase and ß-glucosidase activities. Further, growth temperature was optimal in the range from 33 to 37°C for the cereal strains, whereas the dairy strains showed optimal growth at 40°C. Taken together, adaptation of the various biotypes results in a growth benefit in the particular environment. Acidification and growth tests using either sterile skim milk or a wheat flour extract confirmed these results. Differentiation of these biotypes and their physiological characteristics enables knowledge-based starter culture development for cereal versus dairy products within one species. Biodiversity assessment of dairy- and cereal-derived Lactobacillus helveticus isolates based on genotypic, phenotypic and physiological analyses revealed significant habitat adaptations that are highly relevant for knowledge-based starter culture development.

Background Autophagy-related processes are crucial for maintaining cellular homeostasis in eukaryotic organisms. While alterations of these processes have been strongly linked to specific human disorders, including inflammatory bowel disease, neurodegenerative diseases, and metabolic syndromes, long-term autophagy stimulation appears to be safe and to extend lifespan in model organisms. Several studies indicate that gut microbiota or derived metabolites can modulate host autophagy at the gut mucosa level but also in peripheral organs. Here, we investigated in vitro and in vivo the potential of bacterial species commonly used in food fermentation (ferments) or for their health benefits (probiotics) to modulate host autophagy. Methods We screened 11 bacterial strains (lactobacilli and bifidobacteria) in vitro for autophagy induction in human epithelial cells. The most effective strain identified in vitro was then tested in vivo through long-term dietary supplementation in mice to confirm its pro-autophagic effects in the gut and a distant organ, the retina. Results In vitro screening of the 11 bacterial strains revealed a strain-dependent ability of bacteria to stimulate autophagy in human epithelial cells. The Lactobacillus helveticus strain VEL12193, isolated from cheese, emerged as the strongest autophagy inducer. Long-term dietary supplementation of mice with L. helveticus VEL12193 confirmed the pro-autophagic potential of this strain, as evidenced by autophagy stimulation in the gut mucosa but also at distance, in the retina. Finally, we identified membrane vesicles (MVs) from L. helveticus as a component involved in bacteria-induced autophagy in epithelial and immune cells, with lactate and specific lipid species potentially contributing to this effect. Conclusion In this study, we present evidence that intervention with ferments/probiotics stimulates autophagy in multiple cell types and organs, and we show in vitro that MVs mediate this effect. Additionally, we identify L. helveticus VEL12193 as a promising candidate for the development of healthy-aging strategies.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common type of liver disease worldwide. In a previous study, we confirmed that Lactobacillus helveticus HY7804 (HY7804) improves MASLD by suppressing the expression of mRNAs encoding genes related to hepatic lipogenesis, inflammation, and fibrosis in model mice. Here, we evaluated the ability of HY7804 to restore intestinal barrier function and modulate the gut microbiota, as well as improve MASLD symptoms. Mice fed an MASLD-inducing diet for 7 weeks received HY7804 (109 CFU/kg/day), the Type strain, or positive control (Pioglitazone) during the same period. HY7804 alleviated physiological (p < 0.001) and blood biochemical indicators and reduced MASLD activity scores (p < 0.05) on histological analysis. In addition, HY7804 increased the expression of genes related to fatty acid oxidation (p < 0.001); decreased the expression of apoptosis-related genes (p < 0.001); rescued the expression of tight junction (TJ)-related genes (p < 0.05); and suppressed the expression of pro-inflammatory cytokines and TLR4/MyD88/NF-κB signaling (p < 0.01) in the intestine. Finally, HY7804 modulated the composition of the gut microbiota in MASLD-induced mice. HY7804 increased the abundance of MASLD-suppressive Bacteroidaceae and Bacteroides, which positively correlated with the expression of TJ- and fatty acid oxidation-related genes. By contrast, HY7804 decreased the abundance of bacteria related to the progression of MASLD, including Cloastridaceae, Clostridium, Streptococcaceae, Lactococcus, and Lachnospiraceae, which correlated with intestinal immune responses and MASLD symptoms. In conclusion, L. helveticus HY7804 may be suitable as a functional supplement that alleviates MASLD symptoms and improves intestinal health.