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.

Hot Springs ; Tracheophyta

PURPOSE: The conventional primary therapy for chronic bacterial prostatitis (CBP) is the use of antibiotics. However, the therapy has a low cure rate and long-term use of antibiotics can lead to adverse effects including bacterial resistance. For these reasons, a new therapy for CBP is strongly needed. MATERIALS AND METHODS: To evaluate the anti-inflammatory and antimicrobial effects of selenium-rich hot spring water on CBP, rats were divided into four groups and treatment was administered for four weeks as follows: (1) control (n=8), (2) ciprofloxacin (n=8), (3) selenium-rich hot spring water (n=8), and (4) selenium-rich hot spring water with ciprofloxacin (n=8). Drip infusion of bacterial suspension (E. coli Z17 O2:K1;H-) into Spargue-Dowley rats was then conducted to induce CBP. Four weeks later, the results of prostate tissue and urine culture and histological analysis on the prostate were analyzed in each group. RESULTS: The use of ciprofloxacin, and selenium-rich hot spring water with ciprofloxacin showed statistically significant decreases in bacterial growth and improvements in prostatic inflammation compared with the control group (p<0.05). The selenium-rich hot spring water with ciprofloxacin group showed a statistically significantly lower rate of bacterial growth and and greater improvements in prostatic inflammation than the ciprofloxacin group (p<0.05). CONCLUSIONS: These results suggest that spring water may be an effective material in the treatment of CBP. Notably, the combination treatment of selenium-rich hot spring water and ciprofloxacin has synergistic effects. Therefore, we can suggest that the combination of selenium-rich hot spring water and ciprofloxacin may be effective in the treatment of CBP, and with a higher success rate than ciprofloxacin alone.
Animals ; Anti-Bacterial Agents ; Ciprofloxacin ; Hot Springs ; Inflammation ; Infusions, Intravenous ; Prostate ; Prostatitis ; Rats ; Selenium

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.

Environmental Monitoring ; methods ; Hot Springs ; microbiology ; Legionella ; classification ; isolation & purification ; Microbiological Techniques ; Water Microbiology

OBJECTIVETo study the pollution status of Legionella species in hot spring vacation center and the related factors.

METHODSField surveys were performed in four big hot spring vacation centers of Changping district. Uniform questionnaires was used and colony count was made together with the isolation of Legionella species from hot spring water based on mip gene typing.

RESULTS47 isolates of Legionella pneumophila (Lp) from 87 samples showed 4 serotypes as Lp1, Lp6, Lp12, Lp5 with percent of 57.45%, 21.28%, 14.89%, 6.38% respectively. The hot spring centers controlled the temperature of recycled water between 34-47 degrees C by hot water heating and filtrating system. All the isolates were cultured from the hot water with temperature between 34-44 degrees C: 56.75% (21/37) in high temperature (40-47 degrees C) and 61.90% (26/42) in low temperature (34-39.9 degrees C). There were no statistically significant difference between the high and the low temperature samples (P > 0.05). In the four hot spring vacation centers, the pH value was under control at 6.4-7.3 and the ambient temperature was under control between 26-28 degrees C. The humidity was controlled between 56% -69% relative humidity, which were the best growing conditions for the Legionella species. Disinfectors as chlorine deviratives was used in the four hot spring vacation centers. Though the concentration of chlorine in the water was 0.3-0.5 mg/L, 14.29%-48.00% of the samples were still positive of having Legionella species.

CONCLUSIONThe pollution of Legionella species was considered to be quite serious in the four hot spring vacation centers and the predominant serotype was Lp1. The pH and temperature of the hot spring water, ambient temperature and humidity and the way of heating up the water were the best conditions for the growth of Legionella species in these centers. Because of the high temperature of the hot spring water, chlorine of the disinfector volatilized quickly, affecting the effect of disinfection. The result revealed that water temperature achieving 44 degrees C could have had the effect of prevention.

China ; Disinfection ; Environmental Monitoring ; Hot Springs ; microbiology ; Legionella ; growth & development ; isolation & purification ; Temperature ; Travel ; Water Microbiology

PURPOSE: There have been reports that a higher serum level of selenium is related to a lower incidence of prostate cancer. Serum leptin and IGF-1 are factors related to metabolic syndromes which are also known to be related to prostate cancer, and the higher their levels, the higher the possibility of prostate carcinogenesis. Thus, we measured the serum levels of leptin and IGF-1 in rats that were given selenium-rich hot spring water to drink in an attempt to elucidate the role of selenium on prostate cancer prevention. MATERIALS AND METHODS: A total of twelve white SD rats was included in the study, and they were further divided into a control group (Group 1, n=6) induced to drink normal saline, and those rats that drank selenium-rich hot spring water for 4 weeks (Group 2, n=6). The hot spring water that we used was from the Kum-jin region of Korea, which contains a rich supply of the minerals selenium, vanadium, calcium and magnesium. The concentration of selenium in the hot spring water was 480 microgram/l. The two groups were each given 50 ml/day of normal saline or selenium-rich hot spring water. After the period of the experiment, we drew blood samples from both groups and measured their serum leptin and IGF-1 levels using ELISA. RESULTS: The serum leptin level (pg/ml) was 964.04+/-127.77 in Group 1, and 531.92+/-151.04 in Group 2, allowing us to confirm that it was significantly less in those that drank the selenium-rich hot spring water (p<0.05). Also, the serum IGF-1 levels (pg/ml) were 1,247.72+/-53.85 and 933.03+/-113.69 in Groups 1 and 2, respectively, which again, illustrated the significantly lower level in those rats that were given selenium-rich hot spring water (p<0.05). CONCLUSIONS: Significantly decreased serum levels of leptin and IGF-1 were observed in rats that drank selenium-rich hot spring water. A study of the long-term effect of selenium-rich hot spring water on prostate cancer is necessary; if it is indeed proven to have long-term effects, we believe it will become valuable in clinical practice.
Animals ; Calcium ; Enzyme-Linked Immunosorbent Assay ; Hot Springs ; Humans ; Incidence ; Insulin-Like Growth Factor I ; Korea ; Leptin ; Magnesium ; Minerals ; Prostate ; Prostatic Neoplasms ; Rats ; Selenium ; Vanadium

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

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