BACKGROUND: Although many studies have reported the therapeutic effects of hot spring bathing on various diseases, its influence on healthy individuals is not well understood. Myoban Onsen, a sulfur-rich hot spring in Beppu City, Japan, is traditionally believed to improve skin conditions, relieve fatigue, and promote relaxation. However, scientific verification of these effects, particularly their impact on gut microbiota and related metabolic outcomes in healthy individuals, remains scarce. This study aimed to evaluate the effects of Myoban hot spring bathing on gut microbiota composition and SCFA concentrations in healthy individuals. METHODS: In this study, 16 healthy adult males (n = 16) participated in Myoban hot spring bathing four times over two weeks. Fecal samples were collected before and after the intervention, and 16S rRNA sequencing and gas chromatography-mass spectrometry (GC-MS) were performed to analyze gut microbiota composition and organic acid concentrations. The effects of hot spring bathing were evaluated using the Wilcoxon matched-pair signed-rank test to compare pre- and post-intervention. RESULTS: After Myoban hot spring bathing, there was a significant increase in beneficial gut bacteria, Bifidobacterium, Blautia, and Anaerostipes, compared to pre-bathing (p = 0.0012, p = 0.0103, and p = 0.0017, respectively). Conversely, significant decreases were observed in Parabacteroides, Alistipes, and Oscillibacter (p = 0.0125, p = 0.0215, and p = 0.0125, respectively). Significant increases in SCFAs, including acetic acid, propionic acid, and butyric acid, were observed after Myoban hot spring bathing (p = 0.0067, p = 0.0125, and p = 0.0302, respectively). These findings suggest that Myoban hot spring bathing may benefit healthy adult males. CONCLUSIONS: This study suggests that Myoban hot spring bathing may improve gut health in healthy males. The observed increases in beneficial bacteria and SCFAs indicate a potential contribution to improved health status through modulation of the gut environment. TRIAL REGISTRATION Registration number: UMIN000055229, retrospectively registered.
Humans ; Gastrointestinal Microbiome ; Male ; Hot Springs ; Pilot Projects ; Fatty Acids, Volatile/analysis* ; Adult ; Japan ; Feces/chemistry* ; Bacteria/genetics* ; Young Adult ; Baths ; RNA, Ribosomal, 16S/analysis* ; Middle Aged

Guided by molecular networking, nine novel curvularin derivatives (1-9) and 16 known analogs (10-25) were isolated from the hydrothermal vent sediment fungus Penicillium sp. HL-50. Notably, compounds 5-7 represented a hybrid of curvularin and purine. The structures and absolute configurations of compounds 1-9 were elucidated via nuclear magnetic resonance (NMR) spectroscopy, X-ray diffraction, electronic circular dichroism (ECD) calculations, ¹³C NMR calculation, modified Mosher's method, and chemical derivatization. Investigation of anti-inflammatory activities revealed that compounds 7-9, 11, 12, 14, 15, and 18 exhibited significant suppressive effects against lipopolysaccharide (LPS)-induced nitric oxide (NO) production in murine macrophage RAW264.7 cells, with IC₅₀ values ranging from 0.44 to 4.40 μmol·L^-1. Furthermore, these bioactive compounds were found to suppress the expression of inflammation-related proteins, including inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), NLR family pyrin domain-containing protein 3 (NLRP3), and nuclear factor kappa-B (NF-κB). Additional studies demonstrated that the novel compound 7 possessed potent anti-inflammatory activity by inhibiting the transcription of inflammation-related genes, downregulating the expression of inflammation-related proteins, and inhibiting the release of inflammatory cytokines, indicating its potential application in the treatment of inflammatory diseases.
Penicillium/chemistry* ; Mice ; Animals ; Anti-Inflammatory Agents/isolation & purification* ; RAW 264.7 Cells ; Nitric Oxide/metabolism* ; Hydrothermal Vents/microbiology* ; Macrophages/immunology* ; Molecular Structure ; Nitric Oxide Synthase Type II/immunology* ; Cyclooxygenase 2/immunology* ; Geologic Sediments/microbiology* ; NF-kappa B/immunology* ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology*

Background. The growing resistance of microorganisms to antimicrobial agents is a pressing public health issue. Bioprospecting efforts have mainly focused on well-known environments such as soil and animal gut in search for microorganisms with antibiotic production or antimicrobial activity, or terrestrial ecosystems for endemic plants with bioactive compounds. However, microbial communities thriving in stressed environments such as hot springs, are potential sources of metabolites that can be screened for antimicrobial activity. There is a need for research on bioprospecting of fungi as potential sources of antimicrobials.

Objectives. The study aimed to test the antimicrobial activity of endophytic and rhizospheric fungi associated with soft fern (Christella spp.) and Cinderella weed (Synederella nodiflora) inhabiting a hot spring in Los Baños, Laguna, Philippines.

Methods. A total of 23 endophytic and rhizospheric fungi isolated from soft fern and Cinderella weed were purified and phenotypically identified. These isolates were subjected to agar well diffusion and agar plug diffusion methods as preliminary assays for antimicrobial activity against Bacillus subtilis var. spizizenii (ATCC® 6633™), Staphylococcus  aureus (ATCC® 25923™), four multi-antibiotic resistant Escherichia coli (OT11, OT16, OT18, OT22), and Cladosporium  cladosporioides. Based on the results of the preliminary screening, ethyl-acetate extracts of selected fungal isolates were subjected to broth microdilution assay to determine the minimum inhibitory concentrations (MICs) for antibacterial activity, as well as poisoned food technique to determine the percent mycelial inhibition for antifungal activity. The nearest phylogenetic affiliations of fungal isolates with higher antimicrobial activities were determined.

Results. Ten rhizospheric fungal isolates from Cinderella weed and seven rhizospheric and six endophytic fungal isolates from soft fern were phenotypically identified as Aspergillus, Coniothyrium, Fusarium, Penicillium, Talaromyces,  and Trichoderma species. Ethyl acetate extracts from endophytic fungal isolates UL1 (Trichoderma sp.) and UL2 (Trichoderma sp.) and rhizospheric fungal isolates UR1 (Trichoderma sp.) and UR3 (Trichoderma sp.) showed activity against the test bacteria at 128-256 ?g/mL concentrations. Isolates UL1, UL2, and UR3, which exhibited higher antibacterial activities, were sequenced and confirmed to be most phylogenetically related to Trichoderma  virens. Eleven fungal isolates belonging to Aspergillus spp., Coniothyrium spp., Fusarium spp., Penicillium spp., and Talaromyces spp. demonstrated antagonism against C. cladosporioides. The rhizospheric fungal isolate FCRU4 (Talaromyces sp.), from where ethyl acetate extracts were recovered for testing mycelial inhibition, was confirmed to be most phylogenetically related to Talaromyces islandicus.

Conclusion. Endophytic and rhizospheric fungi asso ciated with Cinderella weed (Synedrella nodiflora) and soft fern (Christella sp.) from a hot spring in Los Baños, Laguna, Philippines have antimicrobial activity.

OBJECTIVES: For the first time, Boliwong, an indigenous community in the Philippines, was surveyed for the prevalence of Cryptosporidium from April to December 2017. METHODS: Cryptosporidium oocysts were detected in samples from the river, creek, and water pumps via immunomagnetic separation techniques, and from human and animal concentrated faecal samples using the modified Ziehl-Neelsen technique. RESULTS: Seven of the 24 water samples (29.2%) were positive for Cryptosporidium, with the highest concentration (0.8 oocyst/L) detected in the creek. Of 35 fecal samples from different animal groups, 8 (21.6%) were positive for Cryptosporidium oocysts. The highest intensity of oocyst shedding was detected in dogs (χ2=8.00). Of the 137 human fecal samples, 39 (28.5%) were infected with Cryptosporidium. In this study, 3 risk factors were found to be associated with infection: (1) location (crude odds ratio [cOR], 16.39; 95% confidence interval [CI], 2.11 to 127.41; p=0.008), (2) drinking water from the natural spring (cOR, 0.29; 95% CI, 0.11 to 0.82; p<0.05), and (3) using an open pit as a sanitary toilet facility (cOR, 2.44; 95% CI, 1.14 to 5.20; p<0.05). When the cOR was adjusted, using an open pit as a sanitary toilet facility remained a significant risk factor of infection (adjusted OR, 0.41; 95% CI, 0.19 to 0.90; p<0.05). CONCLUSIONS: There is a potentially emerging Cryptosporidium zoonosis in Boliwong, Lagawe, Philippines. It is recommended that the toilet facilities and the water system in the community be rehabilitated to avoid any possible disease outbreak. Health education is also needed in the community to maintain proper hygiene and sanitation practices.
Animals ; Cryptosporidium* ; Disease Outbreaks ; Dogs ; Drinking Water ; Epidemiology ; Health Education ; Humans ; Hygiene ; Immunomagnetic Separation ; Natural Springs ; Odds Ratio ; Oocysts ; Philippines* ; Prevalence ; Public Health ; Risk Factors ; Rivers ; Sanitation ; Toilet Facilities ; Water

BACKGROUND: Empirical evidences for efficacy of hot spring (HS) water in inflammatory skin disorders have not been substantiated with sufficient, immunological “hard evidence”. Mageumsan HS water, characterized by its weakly-alkaline properties and low total dissolved solids content, has been known to alleviate various immune-inflammatory skin diseases, including atopic dermatitis (AD). OBJECTIVE: The trial attempted to quantitatively analyze in vitro expression levels of chemical mediators in cutaneous inflammation from HaCaT cell line treated with Mageumsan HS, and suggest the likely mode of action through which it exerts the apparent anti-inflammatory effects in AD. METHODS: Using membrane-based human antibody array kit, customized to include 30 different, keratinocyte-derived mediator proteins, their expression levels (including interleukin [IL]-1, IL-6, IL-8, thymic stromal lymphopoietin, thymus and activation-regulated chemokine, and granulocyte macrophage colony-stimulating factor) were assessed in vitro. Selected key proteins were further quantified with enzyme-linked immunosorbent assay. RESULTS: There was a clear pattern of overall suppression of the mediators, especially those noted for their pro-inflammatory role in AD (monocyte chemoattractant protein [MCP]-1, regulated on activation, normal T cell expressed and secreted, cutaneous T-cell-attracting chemokine, Eotaxin, and macrophage inflammatory protein-1α, etc.). Also, reduced expression of involucrin and cytokeratin 1 was also reduced in the HS-treated group. CONCLUSION: The present study has shown that Mageumsan HS water may exert its effects on inflammatory skin disorders through regulation of proinflammatory cytokines. These evidences are to be supported with further future investigations to elucidate immunological mechanism behind these beneficial effects of HS water in the chronically inflamed skin of AD.
Cell Line ; Chemokine CCL17 ; Chemokine CCL27 ; Cytokines ; Dermatitis, Atopic ; Enzyme-Linked Immunosorbent Assay ; Granulocytes ; Hot Springs* ; Humans ; In Vitro Techniques* ; Inflammation ; Interleukin-6 ; Interleukin-8 ; Interleukins ; Keratins ; Macrophages ; Protein Array Analysis* ; Skin ; Skin Diseases ; Water

OBJECTIVES: For the first time, Boliwong, an indigenous community in the Philippines, was surveyed for the prevalence of Cryptosporidium from April to December 2017.METHODS: Cryptosporidium oocysts were detected in samples from the river, creek, and water pumps via immunomagnetic separation techniques, and from human and animal concentrated faecal samples using the modified Ziehl-Neelsen technique.RESULTS: Seven of the 24 water samples (29.2%) were positive for Cryptosporidium, with the highest concentration (0.8 oocyst/L) detected in the creek. Of 35 fecal samples from different animal groups, 8 (21.6%) were positive for Cryptosporidium oocysts. The highest intensity of oocyst shedding was detected in dogs (χ2=8.00). Of the 137 human fecal samples, 39 (28.5%) were infected with Cryptosporidium. In this study, 3 risk factors were found to be associated with infection: (1) location (crude odds ratio [cOR], 16.39; 95% confidence interval [CI], 2.11 to 127.41; p=0.008), (2) drinking water from the natural spring (cOR, 0.29; 95% CI, 0.11 to 0.82; p<0.05), and (3) using an open pit as a sanitary toilet facility (cOR, 2.44; 95% CI, 1.14 to 5.20; p<0.05). When the cOR was adjusted, using an open pit as a sanitary toilet facility remained a significant risk factor of infection (adjusted OR, 0.41; 95% CI, 0.19 to 0.90; p<0.05).CONCLUSIONS: There is a potentially emerging Cryptosporidium zoonosis in Boliwong, Lagawe, Philippines. It is recommended that the toilet facilities and the water system in the community be rehabilitated to avoid any possible disease outbreak. Health education is also needed in the community to maintain proper hygiene and sanitation practices.
Animals ; Cryptosporidium ; Disease Outbreaks ; Dogs ; Drinking Water ; Epidemiology ; Health Education ; Humans ; Hygiene ; Immunomagnetic Separation ; Natural Springs ; Odds Ratio ; Oocysts ; Philippines ; Prevalence ; Public Health ; Risk Factors ; Rivers ; Sanitation ; Toilet Facilities ; Water

BACKGROUNDSpring water therapies have been used since at least 1550 BC. Despite the growing body of evidence supporting these therapies for a range of conditions, including musculoskeletal, dermatological, respiratory and cardiovascular conditions, they do not currently form part of mainstream healthcare in many countries. The protocol established in this paper aims to support systematic reviews that examine the health outcomes associated with human exposure to regional spring waters, using the Australia and New Zealand context as a case study.

METHODS/DESIGNThe protocol searches for studies in eight health/medical databases, searches three local health/medical journals, and includes forwards and backwards searching. Standard systematic review methods are used including: specifying pre-determined inclusion criteria and data management plans, appraising the studies for bias, and allocation to a hierarchy of evidence.

DISCUSSIONThe protocol supports a review and comprehensive synthesis of the current evidence regarding the health effects of natural spring water, and can be adapted for reviews in other regions. From this evidence, recommendations regarding practice and future research can be made on the therapeutic role of spring water.

BACKGROUND: Balneotherapy, although not a well-established dermatological treatment, is thought to have therapeutic properties for psoriasis and is used as an alternative treatment modality throughout the world. OBJECTIVE: To evaluate the mechanism underlying the therapeutic immunologic effects of thermomineral water. METHODS: A murine model of imiquimod-induced psoriasis-like skin inflammation was used for evaluating the therapeutic effects of balneotherapy with Hae-Un-Dae hot spring mineral water. The clinical improvements were evaluated by a dermatologist. Lesional cytokines, including interleukin (IL)-17A, IL-23, and IL-22, were quantitatively measured by real-time reverse transcriptase polymerase chain reaction. Serum levels of interferon-gamma, IL-4, IL-5, and IL-17A were measured by enzyme-linked immunosorbent assay. T cell proportions in the spleen were evaluated by flow cytometry, and histopathological evaluation of the skin was also performed. RESULTS: The mineral water balneotherapy group showed faster improvement in skin erythema and scales than the distilled water bathing group. A substantial reduction was observed in the lesional mRNA levels of IL-17A and IL-23 in the mineral water group. Serum levels of IL-4 and IL-5 were significantly decreased in the mineral water group but not in the distilled water group. Normalized T cell proportions were observed after bathing. CONCLUSION: Balneotherapy showed immunomodulatory effects in a psoriasis-like murine model. Balneotherapy suppressed lesional IL-23 and IL-17A, which are important cytokines in the pathogenesis of psoriasis. These results suggest that balneotherapy can be used as an effective and safe treatment for psoriasis.
Balneology* ; Baths ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Erythema ; Flow Cytometry ; Hot Springs ; Immunomodulation ; Inflammation ; Interferon-gamma ; Interleukin-17 ; Interleukin-23 ; Interleukin-4 ; Interleukin-5 ; Interleukins ; Mineral Waters ; Psoriasis ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Skin ; Spleen ; Water* ; Weights and Measures

BACKGROUND: Balneotherapy is widely used as an alternative treatment modality for AD. Although the clinical benefit of some mineral waters has been established, their mechanisms of action in alleviating AD are only partly understood. OBJECTIVE: The clinical modification and immunomodulatory or anti-inflammatory effects of mineral water from the Suanbo hot springs on the differentiation and cytokine production of Th1, Th2, and regulatory T cells (Treg) were investigated using spleen, skin tissue, and serum from NC/Nga mice. METHODS: The therapeutic effects of bathing in mineral water in a Dermatophagoides farinae body extract ointment (Dfb ointment)-induced AD mouse model were assessed by measuring the modified Scoring atopic dermatitis (SCORAD) index scores, transepidermal water loss (TEWL), histological and immunohistochemical changes of the skin lesion, serum levels of interferon (IFN)-gamma, interleukin (IL)-4, IL-5 and immunoglobulin E, mRNA expression of IFN-gamma, IL-4 and IL-5 of dorsal skin, and helper T cell differentiation in the spleen. RESULTS: Bathing in mineral water significantly reduced the modified SCORAD index scores, TEWL, epidermal hyperplasia, and inflammatory cell infiltration. IL-4 production and Th2 cell differentiation showed a decreasing tendency with mineral water bathing, but the Th1 cells did not. On the contrary, differentiation to Treg cells was promoted with mineral water bathing. CONCLUSION: Balneotherapy not only has anti-inflammatory activity, but also shows positive effects on cutaneous barrier homeostasis. These results suggest that the favorable effects of balneotherapy may be mediated by modifying the Th2 response, and possibly in part by inducing Treg cell differentiation.
Animals ; Balneology ; Baths ; Cell Differentiation ; Dermatitis, Atopic* ; Dermatophagoides farinae ; Homeostasis ; Hot Springs ; Hyperplasia ; Immunoglobulin E ; Immunoglobulins ; Immunomodulation* ; Interferons ; Interleukin-4 ; Interleukin-5 ; Interleukins ; Mice ; Mineral Waters* ; RNA, Messenger ; Skin ; Spleen ; T-Lymphocytes, Regulatory ; Th1 Cells ; Th2 Cells ; Mineral Waters

OBJECTIVETo compare the detection effect of Legionella pollution in spring water by three methods, namely traditional plating method, fluorescent quantitation PCR method and ethidium monoazide (EMA) fluorescent quantitation PCR method.

METHODSEvery month (except May), we collected 11 water samples from the 5 selected hot spring pools in one hot spring resort in Beijing in 2011. A total of 121 water samples were collected, and then were detected by the above three methods qualitatively and quantitatively.

RESULTSIn our study, the Legionella pollution rate was separately 74.4% (90/121), 100.0% (121/121) and 100.0% (121/121) by the above three methods. The quantitative value of Legionella in the 121 water samples detected by the three methods were around 0.10-216.00 colony-forming units (CFU)/ml, 1.47-1557.75 gene units (GU)/ml and 0.20-301.69 GU/ml, respectively. The median (25th and 75th percentiles) was 75.30 (32.51-192.10) GU/ml, 36.46 (16.08-91.21) GU/ml and 5.30 (0.00-33.70) CFU/ml, respectively. The difference in the quantitative value of Legionella detected by the three methods showed statistical significance (χ(2) = 187.900, P < 0.01). The quantitative value of Legionella detected by fluorescent quantitation PCR method was the highest, followed by the value Legionella detected by EMA-fluorescent quantitation PCR method and traditional plating method.

CONCLUSIONThe sensitivity of the PCR methods was higher than traditional plating method, in detecting Legionella pollution in spring water, especially the EMA- fluorescent quantitation PCR method, which was more suitable for detecting Legionella in water.