Bacillus subtilis

Probiotics are increasingly used in poultry nutrition because of their potential to improve performance and reproductive outcomes. However, there is limited consensus on the effectiveness of different probiotic strains and formulations in broiler breeder hens. This meta-analysis aimed to evaluate the impact of various probiotics on broiler breeder performance and reproductive traits. A comprehensive data set on probiotic supplementation in poultry broiler breeder hens (of different strains) was compiled spanning the time period from 1990-2024. The effect size (Hedge's g) of probiotics was analysed to estimate the random and fixed effects of study variations. Among the rooster strains, the Hubbard strain presented a significant reduction in the number of dead sperm (p < 0.01). Moreover, multistrain probiotics were found to be more effective than single strains (p < 0.01). In hens, probiotics were associated with a significantly lower feed conversion ratio (FCR) (p < 0.001), as were improvements in the fertility and hatchability of settable eggs (p < 0.001). In particular, the addition of probiotics derived from Bacillus subtilis significantly increased fertility and hatchability (p < 0.001). In summary, both single-strain and multistrain probiotics significantly increase various reproductive and performance parameters in broiler breeders, with multistrain probiotics consistently showing superior effects across multiple areas. The inclusion of multistrain probiotics up to 1 g/kg generally yielded positive effects on several parameters in broiler breeder hens. These findings support the inclusion of probiotics, particularly multistrain formulations, to increase reproductive efficiency and performance in broiler breeders, providing a sustainable alternative to antibiotics. © 2025 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.

Probiotics can enhance broiler chicken health by improving intestinal microbiota, potentially replacing antibiotics. They protect against bacterial diseases like necrotic enteritis (NE) in poultry. Understanding their role is crucial for managing bacterial diseases, including NE. This study conducted a meta-analysis to assess the effects of Bacillus subtilis probiotic supplementation on feed conversion ratio (FCR), NE lesion score, and mortality. Additionally, a systematic review analysed gut microbiota changes in broilers challenged with Clostridium perfringens with or without the probiotic supplementation. Effect sizes from the studies were estimated in terms of standardized mean difference (SMD). Random effect models were fitted to estimate the pooled effect size and 95% confidence interval (CI) of the pooled effect size between the control [probiotic-free + C. perfringens] and the treatment [Bacillus subtilis supplemented + C. perfringens] groups. Overall variance was computed by heterogeneity (Q). The meta-analysis showed that Bacillus subtilis probiotic supplementation significantly improved FCR and reduced NE lesion score but had no effect on mortality rates. The estimated overall effects of probiotic supplementation on FCR, NE lesion score and mortality percentage in terms of SMD were -0.91 (CI = -1.34, -0.49; P < 0.001*); -0.67 (CI = -1.11, -0.22; P = 0.006*), and -0.32 (CI = -0.70, 0.06; P = 0.08), respectively. Heterogeneity analysis indicated significant variations across studies for FCR (Q = 69.66; P < 0.001*) and NE lesion score (Q = 42.35; P < 0.001*) while heterogeneity was not significant for mortality (Q = 2.72; P = 0.74). Bacillus subtilis probiotic supplementation enriched specific gut microbiota including Streptococcus, Butyricicoccus, Faecalibacterium, and Ruminococcus. These microbiotas were found to upregulate expression of various genes such as TJ proteins occluding, ZO-1, junctional adhesion 2 (JAM2), interferon gamma, IL12-β and transforming growth factor-β4. Moreover, downregulated mucin-2 expression was involved in restoring the intestinal physical barrier, reducing intestinal inflammation, and recovering the physiological functions

BACKGROUND: Many randomized controlled trials and systematic reviews have evaluated the use of probiotics to treat acute infectious gastroenteritis. However, most probiotic species evaluated in previous large randomized controlled trials are unavailable in Japan. Our objective was to investigate the efficacy of probiotics utilized in Japan for acute gastroenteritis. METHODS: The inclusion criterion was a randomized controlled study that compared probiotics with a placebo to treat children younger than 18 years with acute infectious gastroenteritis. We excluded studies that did not contain the following species available in Japan: Bifidobacterium spp., Lactobacillus acidophilus, Enterococcus faecium, Clostridium butyricum, and Bacillus subtilis and studies in low- or lower-middle-income countries. We searched PubMed, CENTRAL, and Igaku Chuo Zasshi from their inception to November 27, 2022. After the risk of bias assessment, data on diarrhea duration, number of hospitalizations, length of hospital stay, and adverse effects were extracted. RESULTS: Fourteen studies were included in this meta-analysis. Diarrhea lasting longer than 48 h (7 articles, n = 878) was significantly lower in the probiotic group (risk ratio (RR) 0.70, 95 % confidence interval (CI) 0.59-0.83). The duration of diarrhea (14 articles; n = 1761) was 23.45 h (95 % CI 18.22-26.69) shorter in the probiotic group. Duration of hospitalization (6 articles; n = 971) was 17.73 h (95 % CI 6.9-28.56) shorter in the probiotic group. CONCLUSIONS: Although the certainty of evidence is very low, the use of probiotics for acute gastroenteritis in children may improve diarrhea approximately one day earlier. This study was registered with PROSPERO (CRD 42023405559).

Necrotic enteritis (NE) and coccidiosis are among the most prevalent infectious diseases in broiler chickens, contributing to large profitability losses. Bacillus subtilis is a promising direct-fed probiotic to counter various pathogens infection in broiler chickens. Here, we performed a meta-analysis to investigate the effects of B. subtilis on broiler chickens performance. A total of 28 studies were selected according to a PRISMA checklist. Random-effect model and mixed-effect model of meta-analysis were fitted to estimate the overall effects of B. subtilis (BS) treatment compared to either the control group (CON) or NE-infected group (NEinf) as a baseline. Hedges' g effect size and its variance were used as estimators of standardized mean difference (SMD) calculation where the results were presented at a 95% confidence interval (95% CI) of the SMD. Overall, NEinf broiler chickens depressed (P < 0.01) body weight (BW), average daily gain (ADG), and feed intake, and elevated (P < 0.01) feed conversion ratio (FCR). Treatment with BS improved ADG and final BW of NEinf with no difference (P = 0.15) between BS and antibiotics (AB), indicating that they had comparable efficacy to treat NE in broiler chickens. BS supplemented to uninfected CON (BSS) improved (P < 0.01) final BW, ADG, and FCR. Compared to CON, BS, and AB failed to recover the FCR but these treatments decreased (P < 0.01) FCR when compared to the NEinf group with similar efficacy (P = 0.97). As expected, NEinf birds had a higher mortality rate (P < 0.01) and higher lesion score (P < 0.01) compared to CON, and treatment using AB and BS successfully decreased (P < 0.01) the mortality rate and lesion score. Compared to BS, AB was more effective to lower (P = 0.01) mortality rate, but comparable (P = 0.65) to minimize lesion score. To conclude, B. subtilis could be an effective natural additive to replace in-feed antibiotics in broiler chickens challenged with C. perfringens. However, the efficacy to reduce mortality rate was better with antibiotics treatment.

Challenge models are needed to understand the pathogenesis of necrotic enteritis (NE) and provide the basis of evaluating nonantibiotic feed-additive interventions. In the category of nonantibiotic feed additives, the application of probiotics to improve intestinal health and growth performance of broiler chickens in the face of an NE challenge has been well described. However, it is crucial to evaluate the consistency of specific probiotics for mitigating the disease challenge and improving performance. Therefore, a meta-analysis of five independent research trials was conducted with the objective of evaluating the effect of Bacillus subtilis DSM 32315 (probiotic) on body weight gain (BWG), feed conversion ratio (FCR), NE mortality, and lesion score (LS) of broiler chickens challenged with NE. These independent studies were conducted in three countries (the United States, Thailand, and Finland). The statistical analysis used fixed and random effects to estimate the mean effect size (MES) of the difference between NE-challenged birds (control) and NE-challenged probiotic-fed birds and the 95% confidence interval of MES. A meta-regression was performed to evaluate heterogeneity (MES variance) among studies. The statistical analysis was performed using a robust variance estimation strategy with a SAS macro. Probiotic-supplemented birds had a significantly higher BWG (MES = 1.04, P = 0.009) and a significantly lower FCR (MES = -1.39, P = 0.020), NE mortality (MES = -1.15, P = 0.012), and LS (MES = -1.29, P = 0.045). Response variables of BWG (Q = 2.81, P = 0.560) and NE mortality (Q = 5.60, P = 0.354) did not present heterogeneity. Heterogeneity was found for FCR (Q = 10.34, P = 0.035) and LS (Q = 16.13, P = 0.001). Overall, dietary supplementation of B. subtilis DSM 32315 significantly improved BWG and reduced FCR, mortality, and LS in a repeatable large-scale manner.

We aimed to evaluate the efficacy and safety of a spore-forming probiotic blend containing Clostridium butyricum IDCC 1301, Weizmannia coagulans IDCC 1201, and Bacillus subtilis IDCC 1101 for improving bowel function and well-being in adults with functional constipation (FC). In a randomized, double-blind, placebo-controlled trial, 78 adults with FC (Rome IV criteria) received either probiotic blend (n = 40) or placebo (n = 38) daily for 4 weeks. Primary outcomes were changes in weekly spontaneous bowel movements (WSBM) and stool form. Secondary outcomes included physical functioning scores from the 36-Item Short Form Health Survey. The probiotic blend group showed significant improvements in irritant bowel movements (p = 0.0458), incomplete evacuation (p = 0.0374), and abdominal pain before defecation (p = 0.0090). Stool consistency shifted toward normal types (Bristol types 3-4, p = 0.0176). Physical functioning improved only in the probiotic blend group (p = 0.0300). Probiotic blend effectively alleviated symptoms of FC and improved physical well-being. Trial registration: Clinical Research Information Service (CRIS), KCT0010085. Copyright: © 2026 Park et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

BACKGROUND AND AIM: Alterations in gut microbiota after colonoscopy have been observed. Moreover, there is limited consensus regarding the impact of the procedure on gut microbiota. We aim to evaluate whether the probiotic can treat gut microbiota and abdominal symptoms disturbance after colonoscopy. METHODS: A total of 301 participants were randomized into probiotic and placebo groups. The probiotic group received live combined Bacillus Subtilis and Enterococcus Faecium-coated capsules (0.5 g thrice daily for 28 days) postcolonoscopy. The placebo group was given placebo in an identical manner. Fecal samples were collected from participants and symptoms were reported. The intestinal microbiota composition was assessed using 16S rRNA sequencing. RESULTS: Notable differences in α (p = 0.011) and β (p < 0.001) diversity were observed after colonoscopy. Bowel cleansing resulted in the proliferation of Pseudomonas and Klebsiella in the placebo group. However, the probiotic group exhibited a marked enrichment in beneficial probiotics such as Lactobacillales (p = 0.002). Gastrointestinal symptom rating scale (GSRS) showed significant improvements in the probiotic group within the initial week (p = 0.005). Patients in the probiotic group who exhibited symptoms experienced fewer days of abdominal discomfort postcolonoscopy (p = 0.034). Additionally, they exhibited enhanced glucose metabolism (p = 0.040) and a notable increase in Lactobacillales (p = 0.009). In the probiotic group, α and β diversities differed significantly in patients without GSRS improvements, and the concentration of Bacteroides was significantly elevated (p = 0.004). CONCLUSION: Gut microbiota was altered after colonoscopy, especially the proliferation of potentially pathogenic bacteria. Administering probiotics postcolonoscopy leads to beneficial alterations in gut microbiota and associated metabolic pathways, while abdominal discomfort symptoms were improved early. © 2025 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Antibiotics are among the most prescribed medicines in children globally. Antibiotic-associated diarrhoea (AAD) and associated abdominal pain are common side effects, caused by alterations to the intestinal microbiota composition. Most research on probiotic interventions involves prophylactic use of Saccharomyces, lactobacilli or bifidobacteria. Less is known about spore-forming strains administered after AAD onset. Bacillus subtilis HU58™ was found to improve AAD symptoms in adults. This randomised, double-blind, placebo-controlled trial in India investigated the effects of a dietary supplement containing B. subtilis HU58 (2 × 109 cfu/day) versus placebo for seven days on diarrhoea duration (Bristol Stool Scale, BSS), abdominal pain intensity (Visual Analogue Scale, VAS), and overall gastrointestinal wellbeing (adapted Gastrointestinal Restoration Questionnaire, GIRQ; Physician Global Assessment, PGA) in 68 children (1-12 years) with AAD. Between-group comparison of data from each timepoint was conducted for the intention to treat population (Chi-squared test of independence for distribution of BSS types, Wilcoxon rank sum test for mean BSS types, mixed model for repeated measures for VAS, GIRQ and PGA scores). By day 3, 93.5% of the probiotic group had normal BSS types, versus 22.6% in the placebo group ( P < 0.001). Almost all subjects in both groups had normal stool types by day 7. Significantly greater decreases in abdominal pain VAS scores were observed among those receiving the probiotic versus placebo at day 3 (-7.4 [SE 0.5] versus -1.9 [SE 0.3], P < 0.001) and day 7 (-9.1 [SE 0.3] versus -8.5 [SE 0.2], P < 0.001). Greater improvements for both adapted GIRQ and PGA scores assessing GI wellbeing were observed in the probiotic group compared to the placebo group at days 3 and 7 (all P < 0.001). Spore-forming B. subtilis HU58 administered to children after onset of AAD may lead to faster resolution of diarrhoea and associated abdominal pain. Further research with daily clinical assessment and faecal microbiome analysis is warranted. The trial is registered at https://ctri.nic.in/Clinicaltrials (CTRI/2022/02/040138).

Probiotics can confer a beneficial effect on gastrointestinal-related symptoms and their impact on daily life. This randomised, double-blind, placebo-controlled study assessed the efficacy of a novel probiotic strain Bacillus subtilis ATCC 122264 as a probiotic on gas-related symptoms in healthy adults. One hundred healthy adults with functional bloating/distension according to the ROME IV criteria, were randomised in a 1:1 ratio to receive either 5 billion cfu of B. subtilis) or placebo daily for 8 weeks. Intestinal gas symptoms and impact on daily life were assessed weekly by the 17-item Intestinal Gas Questionnaire (IGQ) and the change from baseline was analysed by ANCOVA at 4 and 8 weeks. Post-hoc analyses were conducted on the two parts of the IGQ, scored separately. Sub-group analyses based on sex were also done on the IGQ global and dimension scores as well as the scores from the two individual parts of the IGQ. Safety was assessed by reports of adverse events. B. subtilis did not differ from placebo in the change from baseline to Weeks 4 or 8 in IGQ global score or dimension scores of bloating, flatulence, belching, bad breath, stomach rumbling and difficult gas evacuation ( P > 0.05). In females taking B. subtilis, the severity of belching and flatulence significantly decreased after 4 ( P = 0.046) and 8 weeks ( P = 0.039) respectively, compared to females taking placebo. The impact of flatulence on daily life decreased after 8 weeks in females taking B. subtilis compared to placebo ( P = 0.03). B. subtilis was safe and well tolerated over the 8-week study period. The results suggest that B. subtilis may reduce gas-related symptoms, such as bloating and flatulence, in females. Further studies are needed to confirm the sex-related effects of B. subtilis in populations with gastrointestinal symptoms. Registered at Clinicaltrials.gov: NCT06308146.

Acute rhinosinusitis (ARS) in children may be accompanied by acute otitis media (AOM) which is often associated with bacterial co-infections. These conditions are among the primary reasons that children visit hospitals and require antibiotic treatment. This study evaluated the efficacy of the nasal-spraying probiotics (LiveSpo Navax containing 5 billion Bacillus subtilis and B. clausii spores/5 mL) as a supportive treatment for dual ARS and AOM with otorrhea in a randomized, single-blind, controlled clinical trial. Eighty-two patients (41 per group), aged 1 month to 12 years, received standard care along with nasal spraying of either physiological saline (Control group) or LiveSpo Navax (Navax group), administered three times daily over a 7-day follow-up period. A total of sixty-one patients (30-31 per group) completed the trial. The Navax group experienced 68.00% and 96.77% reductions in nasal congestion (by day 3) and rhinorrhea (by day 7), respectively, which were 2.04 and 1.94-fold higher than the Control group, with odds ratios (OR) of 4.31 and 30.00 (p < 0.05). Endoscopic results indicated 8% and 11% higher reductions in nasal mucopurulent discharge and tympanic membrane hyperemia in the Navax group compared to the Control group. By day 3, compared to day 0, the Navax group exhibited > 1200-fold reduction in Streptococcus pneumoniae and ≥ 4-fold reduction in Haemophilus influenzae concentrations (p < 0.05) in both nasopharyngeal and middle ear fluid samples, whereas the Control group showed no significant reductions. Navax treatment reduced IL-6 by 1.35- to 1.74-fold and TNF-α by 1.17- to 1.45-fold, more effectively than the Control group (p < 0.05). These results suggest that nasal-spray Bacillus spore probiotics, with their ability to reduce bacterial load and modulate immune responses, provide a cost-effective and safe solution for alleviating symptoms of both ARS and AOM in children.Trial registration: ClinicalTrials.gov, Identifier NCT05804123 on April 7, 2023. © 2025. The Author(s).

NANOTECHNOLOGY: is the art and science of dealing with nanoscale particles. This has transformed contemporary dental practices through myriad contributions to biomaterial science. Titanium dioxide nanoparticles procured from Bacillus subtilis, an eco-friendly and biogenic source, can significantly magnify the physiochemical attributes of dental materials. However, postoperative sensitivity is a major drawback of composite restorations. The incorporation of these nanoparticles into dental adhesives can greatly benefit clinical dentistry by resolving this issue. This trial aimed to evaluate the effectiveness of a novel titanium dioxide nanofortified adhesive on the postoperative sensitivity of composite restorations. METHODS: This triple-blind, parallel-group randomized controlled trial was conducted at the Department of Operative Dentistry and Endodontics, School of Dentistry, Islamabad, from May 15, 2023, to November 25, 2023. Participants (n = 60) with Class I and II primary carious lesions with a minimum cavity depth of 3-5 mm were randomly assigned to two groups (n = 30). After obtaining informed consent, the restorative procedure was accomplished using a minimally invasive approach and etch-and-rinse adhesive strategy. In group A, a nanofortified adhesive was used for composite restoration, whereas in group B, an adhesive without nanoparticles was used. Postoperative sensitivity was evaluated using the Visual Analog Scale (VAS) score at follow-up periods: of one day, one week, two weeks and one month. A Chi-square test was used to compare postoperative sensitivity between the two groups. The level of significance was set at p < 0.05. RESULTS: A noteworthy association was observed between sensitivity and the group variable at all four evaluation periods: after one day (p = 0.002), 1 week (p = 0.002), 2 weeks (p = 0.007) and one month. In conclusion, participants who underwent restorative intervention using titanium dioxide nanoreinforced adhesives reported a notable reduction in sensitivity at all time intervals. Hence, the occurrence and severity of postoperative sensitivity are significantly reduced using Bacillus subtilis-procured nanofortified adhesives a

BACKGROUND: Skin aging is one of the most abundant aging-related disorders that can be accelerated by excessive exposure to ultraviolet irradiation. Topically applied fermented skincare ingredients have gained mounting attentions due to their high concentration of various skin nourishing nutrients and bioactive components and low skin irritation potency. AIMS: In the present study, we aim to fully demonstrate the skin-related benefits of a novel extract of Thermus thermophilus and Bacillus subtilis mixed-culture ferment (TBFE). METHODS: TBFE was prepared through an innovative mixed-culture fermentation process. The contents of nutrients and bioactive ingredients were quantified by different methods accordingly. Both in vitro tests and randomized controlled human trial were utilized to further demonstrate multifaceted beneficial effects on human skin, as well as the potential mechanisms. RESULTS: Our results showed that TBFE upregulated the expression of type IV collagen, elastin, aquaporin-3, and dermal-epidermal junction markers, while inhibited production of melanin, in different skin cell models. Moreover, TBFE inhibited the generation of reactive oxygen species and pro-inflammatory mediators induced by ultraviolet irradiation in normal human keratinocytes, while stimulated autophagy in senescent keratinocytes. Results from clinical studies confirmed those in vitro findings, demonstrating that TBFE at 5% and 20% concentration provides anti-aging properties in subjects with sensitive skin, in terms of improving wrinkles, moisturization, and skin lightening. CONCLUSIONS: In summary, we demonstrate that a novel mixed-culture ferment extract has promising anti-aging effects, which may be attributed to anti-oxidation, anti-inflammation, and promotion of autophagy in skin cells. © 2024 The Author(s). Journal of Cosmetic Dermatology published by Wiley Periodicals LLC.

BACKGROUND: Functional gastrointestinal disorders (FGIDs) are common, difficult-to-manage conditions. Probiotics are emerging as a dietary component that influence gastrointestinal (GI) health. We conducted a double-blinded randomised controlled trial of a proprietary strain of deactivated Bacillus subtilis (BG01-4™) high in branched-chain fatty acids (BCFA) to treat self-reported FGID. METHODS: Participants (n = 67) completed a four-week intervention of BG01-4™ (n = 34) or placebo (n = 33). The Gastrointestinal Symptom Rating Scale (GSRS) served as the outcome measure, collected prior to, at two weeks, and at four weeks after completion of the intervention. RESULTS: At four weeks, one of three primary outcomes, constipation in the experimental group, was improved by 33% compared to placebo (15%); both other primary outcomes, Total GSRS and diarrhoea, were significantly improved in both the experimental and placebo groups (32%/26% and 20%/22%, respectively). The pre-planned secondary outcome, indigestion, was improved at four weeks (32%) but compared to the placebo (21%) was not significant (p = 0.079). Exploratory analysis, however, revealed that clusters for constipation (18% improvement, p < 0.001), indigestion (11% improvement, p = 0.04), and dyspepsia (10% improvement, p = 0.04) were significantly improved in the intervention group compared to the placebo. CONCLUSIONS: These initial findings suggest that in people with self-reported FGID, BG01-4™ improves specific symptoms of constipation and related GI dysfunction. Longer-term confirmatory studies for this intervention are warranted. TRIAL REGISTRATION: This study was registered prospectively (25 October 2021) at the Australian New Zealand Clinical Trials Registry (ACTRN12621001441808p).

Influenza virus is a main cause of acute respiratory tract infections (ARTIs) in children. This is the first double-blind, randomized, and controlled clinical trial examining the efficacy of nasal-spraying probiotic LiveSpo Navax, which contains 5 billion of Bacillus subtilis and B. clausii spores in 5 mL, in supporting treatment of influenza viral infection in pediatric patients. We found that the nasal-spraying Bacillus spores significantly shortened the recovery period and overall treatment by 2 days and increased treatment effectiveness by 58% in resolving all ARTIs' symptoms. At day 2, the concentrations of influenza virus and co-infected bacteria were reduced by 417 and 1152 folds. Additionally, the levels of pro-inflammatory cytokines IL-8, TNF-α, and IL-6 in nasopharyngeal samples were reduced by 1.1, 3.7, and 53.9 folds, respectively. Compared to the standard control group, treatment regimen with LiveSpo Navax demonstrated significantly greater effectiveness, resulting in 26-fold reduction in viral load, 65-fold reduction in bacterial concentration, and 1.1-9.5-fold decrease in cytokine levels. Overall, nasal-spraying Bacillus spores can support the symptomatic treatment of influenza virus-induced ARTIs quickly, efficiently and could be used as a cost-effective supportive treatment for respiratory viral infection in general.Clinical trial registration no: NCT05378022 on 17/05/2022. © 2023. Springer Nature Limited.

In the present study, the safety, tolerance and impact of 1×109 cfu Bacillus clausii CSI08, 1×109 cfu Bacillus megaterium MIT411 and a probiotic cocktail containing Bacillus subtilis DE111®, Bacillus megaterium MIT411, Bacillus coagulans CGI314, and Bacillus clausii CSI08 with a total count of 2.0×109 cfu administered daily were assessed as compared with a maltodextrin containing placebo control. A total of 98 study participants received daily doses for 45 days, followed by a washout period of 2 weeks. A questionnaire to capture the incidence and duration of upper respiratory tract, urinary tract and/or gastrointestinal complaints and a diary to capture stool regularity and consistency was kept daily to record compliance throughout the 45 days. Faecal and blood samples were collected for microbiological and haematological analysis at the start and end of the treatment period. The probiotic cocktail significantly decreased the incidence of loose stools throughout the entire study. The recorded respiratory, urinary and gastrointestinal symptoms, defecation frequency and other stool consistency were not influenced. No clinically relevant changes in blood parameters, such as liver and kidney function and no serious adverse events appeared during and after administration. There were no changes in symptoms including sadness, irritability, energy, appetite, tension, stress, sleep, cardiovascular events, aches and pains, and dizziness as determined by a mood questionnaire administered to participants at baseline and at the end of the treatment period. Similarly, the measured inflammatory cytokines, antioxidant levels, cholesterol, triglycerides, free amino acids or minerals remained unaffected. There were no negative changes in alpha or beta diversity of the microbiota with any of the treatment groups. These promising data suggest that these treatments were safe and well tolerated, and further work with larger cohorts are justified to determine the efficacy of these potential probiotics in select demographic groups. Trial registration number with clinicaltrials.gov at NCT04758845.

INTRODUCTION: Antimicrobial resistance and gentamicin-associated toxicity necessitate the development of non-antibiotic adjuvants that enhance aminoglycoside efficacy while reducing required doses. Vitamins B12 and D3 possess physicochemical and biological properties that may influence bacterial resistance pathways. This study aimed to evaluate the modulatory effects of these vitamins on gentamicin against representative bacterial strains using an integrated in vitro-in silico approach. MATERIALS AND METHODS: Eight ATCC Gram-negative and Gram-positive reference strains were evaluated. Antibacterial activity was assessed using agar disk diffusion and broth microdilution assays to determine inhibition zones, minimum inhibitory concentrations (MICs), and MIC fold reduction (MFR). In silico analyses targeted the 30S ribosomal subunit, a cobalamin riboswitch, and a proton motive force (PMF)-related protein to explore mechanistic interactions underlying observed phenotypes. RESULTS: Vitamin B12 significantly enhanced gentamicin efficacy, reducing MICs by up to 8-fold and demonstrating synergistic activity (MFR ≥ 4) against Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis. Conversely, vitamin D3 displayed antagonistic effects (MFR ≤ 0.5) in several Gram-positive strains, increasing gentamicin MICs by up to 16-fold. Computational results indicated cooperative stabilization of gentamicin at the ribosomal target and increased affinity for the cobalamin riboswitch in the presence of vitamin B12, whereas vitamin D3 showed strong binding to a PMF-related protein, consistent with impaired aminoglycoside uptake. CONCLUSION: Vitamin B12 represents a promising dose-sparing adjuvant for gentamicin, while vitamin D3 may compromise aminoglycoside efficacy under the tested conditions. These findings warrant further validation using clinical isolates and biofilm models before considering vitamin-based strategies in aminoglycoside stewardship.

BACKGROUND: The alarming rise of multidrug-resistant (MDR) bacteria, particularly Salmonella spp., has prompted an urgent search for alternative and synergistic antimicrobial strategies. In this study, a novel, green, and multicomponent nanocomposite was synthesized by integrating zinc oxide nanoparticles (ZnO NPs), chitosan (CS), the β-lactam antibiotic ceftazidime (CAZ), and the antidiabetic agent metformin (MTF) straightforward and economical manner. METHODS AND RESULTS: Bacillus subtilis strain ATCC 6633 was used to biosynthesize ZnO NPs, acting as a reliable bio-nanofactory. Various characterization techniques such as FTIR, XRD, TEM, and zeta potential analysis verified the successful integration and structural integrity of the ZnO NPs within the CS nanocomposite containing CAZ and MTF (ZnO/CS/CAZ/MTF). The FTIR spectra confirmed the presence of proteins that act as binding and supportive agents during the biosynthesis process. The produced nanomaterials have a significant positive surface charge of +28.61 mV, which enhances their stability. The particle sizes of the NPs ranged from 9.93 to 17.44 nm. The nanocomposite exhibited strong antibacterial activity against MDR Salmonella enterica subsp., enterica serovar Typhi ATCC 19214, showing a significantly increased inhibition zone of 42 mm and a greatly reduced minimum inhibitory concentration (MIC) value of 8 µg/ml, compared to the separate components. The minimum bactericidal concentration (MBC) value was found to be consistent with the MIC result, emphasizing the potent bactericidal action of the prepared nanocomposite. In silico molecular docking further supported these findings by revealing favorable interactions between the nanocomposite constituents and the outer membrane proteins (OMPs) of Salmonella enterica serovar Typhimurium (PDB ID: 4W4M) and S. typhi (PDB ID: 3UU2). Key interactions included hydrogen bonding, ionic forces, and metal coordination with critical residues. Cytotoxicity assessment using WI-38 lung fibroblast cells revealed an IC₅₀ of 84.26 µg/ml, indicating acceptable preliminary biocompatibility. CONCLUSIONS: The present study demonstrates the novelty of a ZnO-based multicomponent

Eimeria tenella (E. tenella), a pathogenic protozoan parasite, poses a significant threat to the global poultry industry due to its severe impact on intestinal health and growth performance. Given the limitations of conventional anticoccidial drugs, such as the development of drug resistance and chemical residues, it is imperative to explore natural alternative strategies for effective control. In this study, we evaluated the synergistic anti-coccidial effect of pomegranate peel extract (PPE) and Bacillus subtilis (BS). PPE exhibits antioxidant and anti-inflammatory properties, whereas BS, contributes to maintaining intestinal microbial balance. Broiler chickens were challenged with E. tenella and treated with 400 mg/kg PPE in feed and 400 mg/L BS in drinking water, using diclazuril (DIC) as a positive control. The results demonstrated that the PPE+BS combination significantly alleviated cecal lesions, reduced oocyst it mainly parasitizes on the mucosa of the cecum output and mortality, lowered organ indices, suppressed inflammatory responses, and restored body weight gain. The anti-coccidial index (ACI) reached 163, indicating moderate efficacy. Gut microbiota analysis revealed that PPE+BS restored microbial homeostasis by increasing the abundance of Firmicutes, reducing Escherichia coli and Enterococcus, and restoring Micrococcus to levels comparable to uninfected controls. To further explore the potential of the intestinal probiotics, Lactobacillus salivarius (LS) and Lactobacillus curvatus (LC), isolated and identified from chicken feces, achieved comparable anti-coccidial efficacy when combined with PPE, confirming the role of specific probiotics in the observed synergy. In conclusion, the combination of PPE and BS represents an effective and sustainable strategy for controlling avian coccidiosis, primarily through modulation of the gut microbiota and enrichment of beneficial probiotics.

Collagenase and keratinase are two important proteolytic enzymes with recognized applications in biotechnology and medicine, particularly in the enzymatic removal of necrotic tissue and the control of infection. In the present work, a soil isolate of Bacillus subtilis strain AB2 (PX453297.1) was optimized for enzyme production under different nutritional and physicochemical conditions. The enzymes were recovered by ammonium sulphate precipitation and dialysis, examined by SDS-PAGE and zymography, and further assessed for pH and temperature optima, stability, the influence of metal ions, and kinetic parameters. Maximum collagenase activity (4.41 ± 0.22 U/ml) was observed at 37°C and pH 7.5 in a glucose-peptone medium, whereas keratinase production was enhanced between 37 and 40°C at pH 7.5 in lactose-peptone medium. Protein bands of approximately 55 and 33 kDa were detected, representing 6.2- and 5.5-fold purification. Collagenase showed an alkaline optimum (pH 10.0, 37-45°C) with Km 0.31% and Vmax 1.92 U/ml, while keratinase exhibited dual optima (pH 3.0 and ~7.0) with Km 0.27% and Vmax 0.84 U/ml. Biofilm assays revealed that collagenase reduced pre-formed biomass by 62-68% and viable counts by 1.1-1.7 log₁₀, clearly outperforming keratinase (41-57%, 0.7-1.2 log₁₀). When combined with conventional antibiotics, both enzymes potentiated activity, with notable synergy between collagenase and oxacillin against Staphylococcus aureus (FICI 0.31-0.37), ciprofloxacin against Pseudomonas aeruginosa (FICI 0.37-0.50), and meropenem against Klebsiella pneumoniae (FICI 0.28-0.44). These results indicate that B. subtilis AB2 produces collagenase and keratinase with distinct biochemical characteristics and strong antibiofilm properties, underscoring their promise as adjuncts in chronic wound care as well as in industrial applications.

Six new polyketides, including three naphthalene-types (1-3) and three γ-lactones-types (4-6), along with 14 known polyketides (7-20), were isolated from a pathogen endophytic fungus Phomopsis sp. XP-8 of Eucommia ulmoides Oliver. Their structures were elucidated by spectrometric analysis using MS and NMR data. The absolute configurations were established using statistical comparative analysis of the experimental ECD data, in conjunction with quantum mechanical calculations. All the compounds and crude extracts were evaluated for their antibacterial activity. Compounds 2, 3, 6, 7, 8, and 10 exhibited moderate inhibition on three gram-positive bacteria, Staphylococcus aureus, Bacillus subtilis, and Bacillus thuringiensis. Synergistic antibacterial interaction experiments showed that compounds 1, 2, 3, 6, 7, 8, and 19 with antibiotics can greatly enhance the inhibition rate against bacteria. The target screening and molecular docking of these compounds were also carried out, with further verification of molecular dynamics, revealing that the compounds exert antibacterial activity by targeting both the Fur-regulated basic protein (FbpB) and drug resistance protein D (EmrD), exhibiting synergy with antibiotics by inhibiting the DNA polymerase III β-subunit resulting in enhanced efficacy.

Starch digestion induces postprandial hyperglycemia, which can be modulated by polyphenols and polysaccharides via α-amylase inhibition. Current research focuses on individual α-amylase inhibitors, neglecting the activity changes of combined compounds induced by structural differences. This study explored the inhibitory effect and mechanism of the epigallocatechin gallate (EGCG) and exopolysaccharides from Bacillus subtilis STB22 (BS-EPS) complex on porcine pancreatic α-amylase (PPA). The results showed that EGCG (30 mg/mL) and BS-EPS (0.1 mg/mL) individually inhibited PPA by 44.47 % and 21.55 %, respectively, while their complex exhibited enhanced inhibition (72.95 %). The combination index ≤1 confirmed a synergistic and additive inhibitory effect. Mechanistic studies revealed that EGCG and BS-EPS acted as non-competitive and mixed-type inhibitors of PPA, respectively. When combined, they formed a monolayer adsorption complex that exhibited mixed-type inhibition on PPA and statically quenched PPA fluorescence via ternary complex formation. These findings provide a theoretical foundation for developing novel α-amylase inhibitors.

AIMS: Chelating agents are metal-sequestering compounds with antibacterial properties suitable for commercial and therapeutic applications. This study investigated the involvement of metal restriction and membrane disruption in the antibacterial mode of action of three chelators. METHODS AND RESULTS: The antibacterial, metal sequestration, and membrane disruptive effects of ethylenediaminetetraacetic acid, diethylenetriamine pentamethylene phosphonic acid, and fusaric acid were examined across five bacterial species. ICP-MS was used to determine the impact on bacterial metal composition, while RT-qPCR of selected genes allowed evaluation of changes in cellular responses to intracellular metal depletion. Mutants defective in metal import and export machinery were also examined to validate processes critical for resistance. Chelator-mediated disruption of membranes was investigated using 1-N-phenylnapthylamine and propidium iodide. Finally, the capacity of two of the chelators to potentiate the activity of ampicillin, chloramphenicol, tetracycline, and three aminoglycosides was assessed in chequerboards. The results show that these chelators restrict access to iron, zinc, manganese, and calcium to varying degrees in these bacterial species, reflecting important differences in envelope architectures and metal handling capabilities. CONCLUSIONS: This study shows that all three chelators behave differently in restricting metal access and possess antibacterial properties that often act synergistically in combination, notably with other antimicrobials. © The Author(s) 2025. Published by Oxford University Press on behalf of Applied Microbiology International.

CONTEXT: The global rise in bacterial resistance to existing antibiotics has led to increased mortality rates and escalating healthcare costs, underscoring the urgent need for new classes of antibacterial agents that can act synergistically with current antimicrobials. OBJECTIVE: This study aimed to isolate bioactive compounds from Satureja hortensis L. (Lamiaceae) and evaluate their effects in combination with ciprofloxacin (CIP), ampicillin (AMP), gentamicin (GEN), and tetracycline (TET). MATERIALS AND METHODS: The adjuvant potential of naringenin (N), 5,6-dihydroxy-7,3',4'-trimethoxyflavone (TMF), cilicione-a (Cil), rosmarinic acid (Rs), rosmarinic acid methyl ester (MRs), and fulgidic acid was evaluated by determining the minimum inhibitory concentrations (MICs) of selected antibiotics in presence of sub-inhibitory concentrations of these compounds. The assays were performed against ten bacterial strains using the twofold broth microdilution method. Test samples were prepared in 96-well microtiter plates, incubated at 37 °C for 18 h, and dimethyl sulfoxide was used as the solvent control. RESULTS: N, Rs, and MRs reduced the MIC value of GEN from 0.78 µM to 0.097 µM against the drug-resistant Escherichia coli AG100 strain. Additionally, the combinations of N and MRs with CIP reduced its MIC from 3.125 µM to 0.39 µM against Klebsiella pneumoniae. A MIC reduction (from 50 µM to 12.5 µM) was observed against the methicillin- and oxacillin-resistant Staphylococcus aureus strain when N, Cil, Rs, and MRs were combined with AMP. The MIC changed from 0.125 µM to 0.03125 µM against Bacillus subtilis when N and TMF were combined with GEN. DISCUSSION AND CONCLUSIONS: These findings highlight the potential of such combinations to enhance antibiotic efficacy, lower the required dosages, and contribute to mitigating the emergence of antimicrobial resistance.

OBJECTIVES: Antibiotic resistance towards penicillin has been attempted to counter by chemically modifying ampicillin through the conjugation with silver nanoparticles (AgNPs). The current study optimizes the conditions for synthesizing and characterizing AgNP-ampicillin to quantify the conjugation extent, evaluate the antibacterial efficacy, and explore the underlying antibacterial mechanisms. MATERIALS AND METHODS: AgNPs were synthesized from silver nitrate by chemical reduction method, silica-coated with tetraethyl orthosilicate (TEOS) and amine functionalized by (3-aminopropyl) triethoxysilane (APTES), which was then conjugated with ampicillin via the carbodiimide chemistry. UV-visible spectroscopy and DLS were employed to confirm the synthesis of AgNPs, while FT-IR and TGA were used to confirm ampicillin functionalization and conjugation, and SEM and EDX spectroscopy provided morphological insight. Microbial assays were conducted against Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa to determine the inhibition zones, MIC, MBC, MPC, MBIC, MBEC, FIC index, and time-dependent efficacy of AgNP-ampicillin. Cytotoxicity was assessed on Vero cells while molecular docking was performed using AutoDock Vina. RESULTS AND DISCUSSIONS: The synthesized conjugates demonstrated an approximate conjugation efficiency of 57.7%, with four ampicillin molecules conjugated per AgNP. The AgNP-ampicillin conjugates exhibited enhanced antibacterial activity against the studied clinical isolates compared to AgNPs or ampicillin alone, as evidenced by significant differences in inhibition areas in One-way ANOVA (F=18-25.68, p<0.05), while Tukey's post-hoc analysis suggested synergistic effects. AgNP-ampicillin demonstrated enhanced bacteriostatic and bactericidal activity against both planktonic and biofilm-forming cells with mutant prevention ability, and upto 1.25 times faster bacterial elimination compared to ampicillin and AgNPs alone. Synergistic effects of AgNP-ampicillin were confirmed by an FIC index (≤0.5), and effective protection of ampicillin from β-lactamase degradation was established through molecular docking. Cytotoxicity testing

Biological green synthesis of silver nanoparticles (AgNPs) offers potent antimicrobial properties and provides a promising approach against drug-resistant microbes. The present study aims to synthesize AgNPs using Mung bean (Vigna radiate) aqueous extract and the probiotic Bacillus clausii, and to evaluate their antibacterial activity individually and in combination against several bacterial strains. Synthesized AgNPs were characterized by ultraviolet-visible spectroscopy (UV-vis) and scanning electron microscopy (SEM). V. radiata AgNPs were revealed at 450 nm, with spherical to oval shapes ranging in size from 15.9 to 23.0 nm; probiotic AgNPs were observed at 425 nm, also with a spherical shape ranging in size from 16.4 to 23.7 nm. The antibacterial assay was performed using the agar well diffusion method against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli. Results indicated that V. radiata aqueous extract and the probiotic solution did not exhibit any effect, whereas synthesized V. radiata and probiotic AgNPs showed antibacterial activity against all tested bacteria. AgNPs possessed higher antibacterial activity than individual AgNPs when used in combination against almost all bacteria studied. Therefore, it is suggested that using natural antimicrobial agents to synthesize NPs could serve as an eco-friendly alternative to antibiotics.

BACKGROUND: Organic fertilizers are safer and more eco-friendly than chemical fertilizers; hence, organic fertilizers can be used to support sustainable farming. The effects of PGPRs are manifold in agriculture, especially in monoculture crops, where the soil needs to be modified to increase germination, yield, and disease resistance. The objective of this study was to assess the effects of PGPRs combined with fertilizer on the yield and productivity of canola. Canola was chosen for its global importance as an oilseed crop and its responsiveness to soil amendments, making it ideal for evaluating the synergistic effects of PGPRs and fertilizers on yield and soil health. METHODOLOGY: This research, which was carried out over two years, was aimed at establishing the effectiveness of PGPRs together with organic and inorganic fertilizers on canola yields and was performed with a two-factorial RCBD design under field conditions. We applied Azotobacter salinestris and Bacillus subtilis with biochar, compost, animal manure, poultry manure, and NPK fertilizer. Insect pest management and other agronomic practices were carried out to maintain the experiment. RESULTS: Canola yield and agronomic traits were enhanced by the combination of Bacillus subtilis with the fully recommended N: P:K ratio (140:55:40 kg/ha). Additionally, the application of Bacillus subtilis with biochar at 2 tons/ha improved the yield and quality of canola, as well as the structure and nutrient regulation of the soil. CONCLUSION: In light of these results, we recommend the application of Bacillus subtilis to canola seeds along with either 2 t/ha biochar or the entire recommended dose of N: P:K (140:55:40 kg/ha). These strategies are sustainable and help producers and the environment increase the productivity of canola. Combining PGPRs with fertilizers for canola enhances nutrient efficiency, promotes sustainable growth, and boosts stress resilience, addressing agricultural and environmental challenges. CLINICAL TRIAL NUMBER: Not Applicable. © 2025. The Author(s).

BACKGROUND/AIMS: The effects of probiotic supplementation on Helicobacter pylori (H. pylori) eradication therapy are not completely understood. In this study, we investigated the effects of continuous probiotic administration on eradication rates, recrudescence, and symptom response following completion of a course of H. pylori therapy. METHODS: This prospective, randomized, double-blind placebo-controlled trial was performed between June 2018 and 2020. Twohundred seventy patients who received a standard triple regimen for H. pylori eradication, were included in the study. Participants were randomized to receive a probiotic as adjunctive therapy (Enterococcus faecium 4.5×108 and Bacillus subtilis 5.0×107; Medilac-S®, Hanmi Pharmaceuticals, Seoul, Korea) or a placebo (one tablet thrice daily) for 28 days, following H. pylori eradication. Participants who showed successful eradication underwent a repeat 13C-urea breath test after 6 months. RESULTS: Eradication rates in the probiotic and placebo groups were 77.1% and 72.4%, respectively (P=0.48) using per-protocol analysis. Using intention-to-treat analysis, eradication rates were 67.4% and 65.9%, respectively (P=0.43). Of 149 patients who were followed-up after 6 months, four patients had recrudescence (2.7%). Recrudescence rates did not differ between the probiotic and placebo groups. Of the 76 patients who had non-ulcer dyspepsia, 60 (78.9%) showed symptom resolution after 6 months. This beneficial effect was most pronounced in patients with postprandial distress syndrome (P=0.02). CONCLUSIONS: Consecutive probiotic supplementation following H. pylori eradication therapy did not increase eradication rates or decrease recrudescence rates.

A 7-week feeding trial was conducted to evaluate the combined effects of propionic acid (PA, 5 or 10 g/kg) and a multi-strain Bacillus spp. (Bacillus subtilis IS02 (accession no. JN856456) and B. licheniformis IBRC-M 11,319) (1.7 × 107 CFU/g) probiotic in a plant protein source (PP)-rich diet (∼70% of dietary protein derived from PP sources) on performance of Asian sea bass (Lates calcarifer) fry (initial body weight 2.97 ± 0.11 g). In this regard, six isoproteic (∼48%) diets were formulated as follows: a control (without supplementation of the additives); probiotic (only contained Bacillus spp. mixture); 5 g PA/kg diet; 10 g PA/kg diet; probiotic + 5 g PA/kg diet, and probiotic + 10 g PA/kg diet. Specific growth rate in fish fed with 10 g PA/kg (2.84 ± 0.1%) and diets contained blends of probiotic and PA (2.76 ± 0.19% in probiotic + 5 g PA, and 2.79 ± 0.04% in probiotic + 10 g PA) was better than in the control (2.45 ± 0.1%) (P < 0.05). Feed conversion ratio in fish fed with 10 g PA/kg (0.92 ± 0.12) and diets contained blends of probiotic and PA (0.94 ± 0.06 in probiotic + 5 g PA and 0.91 ± 0.02 in probiotic + 10 g PA) was better than in the control (1.24 ± 0.08) (P < 0.05). Digestive enzymes including α-amylase, total alkaline proteases, and bile salt dependent lipase activities improved in fish fed diets contained additives. The activity of glutathione-S-transferase and glutathione reductase enhanced in the liver of fish fed diets contained additives. The relative abundance of lysozyme, interleukin 1β, and insulin-like growth factor-1 genes mRNA transcript showed multifold increase in the liver of fish fed with the 10 g PA/kg and diets contained blends of probiotic and PA (P < 0.05). By considering the above mentioned results, supplementing a PP-rich diet with 10 g PA/kg diet or combination of PA and a mixture of Bacillus spp. probiotic recommended for L. calcarifer. © 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

Pexidartinib is a kinase inhibitor that induces tumor response and improvements in symptoms and functional outcomes in adult patients with symptomatic tenosynovial giant cell tumor (TGCT). A population pharmacokinetic (PK) model for pexidartinib and its metabolite, ZAAD, was developed, and effects of demographic and clinical factors on the PK of pexidartinib and ZAAD were estimated. The analysis included pooled data from 7 studies in healthy volunteers (N = 159) and 2 studies in patients with TGCT or other solid tumors (N = 216). A structural 2-compartment model with sequential zero- and first-order absorption and lag time, and linear elimination from the central compartment adequately described pexidartinib and ZAAD PKs. Clearance of pexidartinib was estimated at 5.83 L/h in a typical patient with reference covariates (male, non-Asian, weight = 80 kg, creatinine clearance ≥90 mL/min, aspartate aminotransferase ≤80 U/L, and total bilirubin ≤20.5 μmol/L). In the covariate analysis, Asians and healthy subjects had modestly lower pexidartinib exposure (21% decrease each) in terms of steady-state area under the curve values from 0 to 24 hours (AUC0-24,ss ). Effects of body weight, sex, and hepatic function parameters on pexidartinib AUC0-24,ss were generally <20%. Patients with TGCT with mild renal impairment were predicted to have approximately 23% higher AUC0-24,ss than those with normal renal function. The effects of covariates on ZAAD exposure were similar to those on pexidartinib. These results indicate small and generally clinically nonmeaningful effects of patient demographic and clinical characteristics on pexidartinib and ZAAD PK profiles. © 2020 Daiichi Sankyo, Inc. The Journal of Clinical Pharmacology published by Wiley Periodicals LLC on behalf of American College of Clinical Pharmacology.

Context: Citrus unshiu Markovich (Rutaceae) peel is known to contain high concentrations of flavonoids and exerts pharmacological effects on antioxidant, anti-inflammation, allergies, diabetes and viral infections. Objective: Very little is known about potential activity of fermented dried Citrus unshiu peel extracts (FCU) using Bacillus subtilis, as well as its mechanism of action. We investigated the effects of FCU on the anti-inflammatory activities in murine macrophages and moisturizing effects in human keratinocytes. Materials and methods: We isolated the Bacillus subtilis from Cheonggukjang and FCU using these Bacillus subtilis to prepare samples. The cells were pre-treated with various extracts for 2 h and then induced with LPS for 22 h. We determined the NO assay, TNF-α, IL-6 and PGE2 in RAW 264.7 ells. The expression of SPT and Filaggrin by FCU treatment was measured in HaCaT cells. Result: We found that two types of FCU highly suppressed LPS-induced nitric oxide (NO) without exerting cytotoxic effects on RAW 264.7 cells (21.9 and 15.4% reduction). FCU inhibited the expression of LPS-induced iNOS and COX-2 proteins and their mRNAs in a concentration-dependent manner. TNF-α (59 and 30.9% reduction), IL-6 (39.1 and 65.6% reduction), and PGE2 secretion (78.6 and 82.5% reduction) were suppressed by FCU in LPS-stimulated macrophages. Furthermore, FCU can induce the production of hyaluronic acid (38 and 38.9% induction) and expression of Filaggrin and SPT in HaCaT keratinocyte cells. Discussion and conclusion: FCU potentially inhibits inflammation, improves skin moisturizing efficacy, and it may be a therapeutic candidate for the treatment of inflammation and dry skin.