Isolation and identification of Balneatrix alpica from patient′s blood and hot spring water / 中华微生物学和免疫学杂志

Objective:To identify and characterize two Balneatrix alpica strains isolated from a patient′s blood sample (strain X117) and the natural hot spring water in the patient′s residential district (strain GN-1), and to provide experimental evidence for the pathogenic diagnosis of clinical infection caused by this rare pathogen. Methods:Biochemical phenotypic identification, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), 16S rRNA gene sequencing, phylogenetic analysis, single-nucleotide polymorphism (SNP) analysis, and genome-wide analysis were performed to accurately determine the taxonomic status of the isolates X117 and GN-1 by using Balneatrix alpica DSM 16621 T as a reference. Microdilution broth method was used to test their antimicrobial susceptibility. The virulence genes carried by them were annotated and analyzed using the virulence factor database (VFDB). Results:Strains X117 and GN-1 formed light yellow or tan colonies with mottled surfaces on Columbia blood agar and chocolate agar plates after 4 d of culture. They were Gram-negative rods and positive for oxidase and indole tests, which were consistent with the characteristics of Balneatrix alpica DSM 16621 T. The phylogenetic analysis based on the 16S rRNA gene showed that the isolates X117 and GN-1 were both Balneatrix alpaca. The average nucleotide identity (ANI) values between the two isolates and Balneatrix alpica DSM 16621 T were 98.44% and 98.41%, respectively, and the digital DNA-DNA hybridization (dDDH) values were both 87.1%. The SNP distance between the two strains was 13, indicating that X117 and GN-1 might belong to the same clone. The antibiotic susceptibility testing showed that all of the three Balneatrix alpica strains were sensitive to the commonly used antibiotics against Gram-negative rods. The virulence genes carried by the three Balneatrix alpica strains were mainly involved in adhesion, invasion, flagella and biofilm formation. Conclusions:This study identified a case of bloodstream infection caused by Balneatrix alpica which was closely related to natural hot spring water. Natural hot spring water migh be an important source of clinical infections caused by this species.

Identification of Gram Positive and Negative Bacteria Microflora Isolated from Hot Water Spring in Bentong, Pahang / Jurnal Sains Kesihatan Malaysia

A study on the isolation and identification of bacterial flora from hot spring was carried out from Bentong hot springwater, Pahang. In general, many of the bacteria identified from the hot spring were from Bacillus sp. due to its abilityto adapt to high temperature mainly by forming endospore. The objective of this study was to identify bacteria otherthan Bacillus sp. that were able to survive this hot spring. Three hot spring pools were identified as A, B and C. Thetemperatures were ranging from 36°C to 52°C and their pH values were between 7.88-8.65. The result shows thattwo species of Gram positive cocci and three species of Gram negatives were isolated. The two Gram positives wereStreptococcus sp. and Staphylococcus sp. The three species of Gram negative rods were identified as Pseudomonas sp.,Klebsiella sp. and Proteus sp. In conclusion, non-sporulating species of bacteria other than Bacillus sp. have adaptedto survive in hot spring

Investigation of Immune-Regulatory Effects of Mageumsan Hot Spring via Protein Microarray In Vitro

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.

Immunoinflammatory Regulation Effects of Korean Hot Spring Water / 日本温泉気候物理医学会雑誌

<b>Background: </b>The beneficial clinical effects of Korean hot spring spa therapy, as well as their underlying mechanisms are still poorly understood. We performed a series of clinical and laboratory investigations for better understanding of the clinical effects as well as possible mechanisms of their beneficial effects.<BR><b>Methods:</b> HaCaT cells were prepared and treated with TLR agonist in the presence or absence of HS water for quantification of IL-6, IL-8, GM-CSF, and TNF-<i>α</i> levels. The serum levels of IFN-ɤ, IL-4, IL-5, and IgE were measured. CD4⁺ naïve cells were allowed to polarize into Th1, Th2, Th17, and Treg cells, and CD4⁺ and CFSE⁺ cells were measured for the degree of proliferation. Total RNA from the lesional skin was transcribed into cDNA using a reverse transcription system, and RT-PCR was performed subsequently. Confocal microscopy and RT-PCR were utilized to evaluate the target skin localization of Th cell subsets and associated inflammatory cytokine milieu.<BR><b>Results: </b>Treatment with agonists of TLR 1 through 6 induced attenuation of cytokine production in the exposure to HS water. HS water suppressed the proliferation of Th1, Th2, and Th17 cells with anti-CD3 stimulation, while proliferation and differentiation to Treg cells were promoted under HS water treatment. On RT-PCR of the lesional skin, thymic stromal lymphopoietin (TSLP) mRNA decreased dramatically after bathing with HS. IL-33 mRNA decreased markedly in HS water group as compared to control group. Foxp3 mRNA expression, same as in confocal microscopic finding, showed tendency to increase more in HS.<BR><b>Conclusions:</b> HS water suppressed the proliferation of Th1, Th2, and Th17 cells. In contrast, proliferation and differentiation to Treg cells were promoted under HS water treatment. These results indicate that HS water may affect the distribution of the helper T cells in the immune response, by suppressing the polarization of the Th1, Th2, and Th17 cells. Also, APC induced TNF-<i>α</i> and IL-6 levels were reduced in the presence of HS water. These results showed that TLR-triggered inflammatory responses in APCs might also be modulated under HS water treatment. Overall, our findings suggest that HS spa therapy could be an effective and safe modality for the management of adult AD.

Immunoinflammatory Regulation Effects of Korean Hot Spring Water / 日本温泉気候物理医学会雑誌

Background: The beneficial clinical effects of Korean hot spring spa therapy, as well as their underlying mechanisms are still poorly understood. We performed a series of clinical and laboratory investigations for better understanding of the clinical effects as well as possible mechanisms of their beneficial effects. Methods: HaCaT cells were prepared and treated with TLR agonist in the presence or absence of HS water for quantification of IL-6, IL-8, GM-CSF, and TNF-α levels. The serum levels of IFN-ɤ, IL-4, IL-5, and IgE were measured. CD4+ naïve cells were allowed to polarize into Th1, Th2, Th17, and Treg cells, and CD4+ and CFSE+ cells were measured for the degree of proliferation. Total RNA from the lesional skin was transcribed into cDNA using a reverse transcription system, and RT-PCR was performed subsequently. Confocal microscopy and RT-PCR were utilized to evaluate the target skin localization of Th cell subsets and associated inflammatory cytokine milieu. Results: Treatment with agonists of TLR 1 through 6 induced attenuation of cytokine production in the exposure to HS water. HS water suppressed the proliferation of Th1, Th2, and Th17 cells with anti-CD3 stimulation, while proliferation and differentiation to Treg cells were promoted under HS water treatment. On RT-PCR of the lesional skin, thymic stromal lymphopoietin (TSLP) mRNA decreased dramatically after bathing with HS. IL-33 mRNA decreased markedly in HS water group as compared to control group. Foxp3 mRNA expression, same as in confocal microscopic finding, showed tendency to increase more in HS. Conclusions: HS water suppressed the proliferation of Th1, Th2, and Th17 cells. In contrast, proliferation and differentiation to Treg cells were promoted under HS water treatment. These results indicate that HS water may affect the distribution of the helper T cells in the immune response, by suppressing the polarization of the Th1, Th2, and Th17 cells. Also, APC induced TNF-α and IL-6 levels were reduced in the presence of HS water. These results showed that TLR-triggered inflammatory responses in APCs might also be modulated under HS water treatment. Overall, our findings suggest that HS spa therapy could be an effective and safe modality for the management of adult AD.

PT2-3Immunoinflammatory regulation effects of Korean hot spring water / 日本温泉気候物理医学会雑誌

  Spa therapy has long been utilized as one of safe, effective and long-term alternative therapies of various dermatologic diseases in Korea. Among them are pruritic, xerotic, immunoinflammatory, infectious, ulcerative skin diseases. There are about 400 spas in South Korea and many of them are recognized as having specific therapeutic effects. But, the beneficial clinical effects of spa therapy as well as their underlying mechanisms are still poorly defined and understood. In Korea, the area of distribution of hot springs coincide with areas of granite and are distant from geothermal heat energy related to volcanos. Natural hot spring water in South Korea can be classified into four main hydrochemical groups, i.e., Ca(Na)-HCO₃, Na(Ca)-HCO₃, Na(Ca)-Cl, and acidic Ca-HCO₃ type. They are mostly alkaline in nature and have low chemical constituents. We performed a series of clinical and laboratory studies to have better understanding of the clinical effects as well as possible mechanisms of their beneficial effects of several hot springs having different locations and mineral concentrations. They are Haeundae(Na-Cl type), Seokmo-do(Na-Cl, Ca-Cl type), Suanbo(Na-HCO₃ type), Baekam(Na-HCO₃ type), and Dukku(Na-Cl type) areas. Utilizing human, mice models, and keratinocyte cell lines, we evaluated the effects of each hot spring water on the changes of clinical symptoms of induced dermatitis and their skin barrier functions, skin infiltrating cells and related histologic findings, proinflammatory cytokine production, helper T cell subset differentiation and proliferation. We found that studied hot spring waters, in general, have the capacity to improve skin inflammation of atopic dermatitis and tendencies to downregulate the production of skin inflammatory cytokines and to increase the Foxp3⁺ Treg cell population while to decrease the Th1, Th2, and Th17 populations.

PT2-3 Immunoinflammatory regulation effects of Korean hot spring water / 日本温泉気候物理医学会雑誌

  Spa therapy has long been utilized as one of safe, effective and long-term alternative therapies of various dermatologic diseases in Korea. Among them are pruritic, xerotic, immunoinflammatory, infectious, ulcerative skin diseases. There are about 400 spas in South Korea and many of them are recognized as having specific therapeutic effects. But, the beneficial clinical effects of spa therapy as well as their underlying mechanisms are still poorly defined and understood. In Korea, the area of distribution of hot springs coincide with areas of granite and are distant from geothermal heat energy related to volcanos. Natural hot spring water in South Korea can be classified into four main hydrochemical groups, i.e., Ca(Na)-HCO3, Na(Ca)-HCO3, Na(Ca)-Cl, and acidic Ca-HCO3 type. They are mostly alkaline in nature and have low chemical constituents. We performed a series of clinical and laboratory studies to have better understanding of the clinical effects as well as possible mechanisms of their beneficial effects of several hot springs having different locations and mineral concentrations. They are Haeundae(Na-Cl type), Seokmo-do(Na-Cl, Ca-Cl type), Suanbo(Na-HCO3 type), Baekam(Na-HCO3 type), and Dukku(Na-Cl type) areas. Utilizing human, mice models, and keratinocyte cell lines, we evaluated the effects of each hot spring water on the changes of clinical symptoms of induced dermatitis and their skin barrier functions, skin infiltrating cells and related histologic findings, proinflammatory cytokine production, helper T cell subset differentiation and proliferation. We found that studied hot spring waters, in general, have the capacity to improve skin inflammation of atopic dermatitis and tendencies to downregulate the production of skin inflammatory cytokines and to increase the Foxp3+ Treg cell population while to decrease the Th1, Th2, and Th17 populations.

Identification of Bacterial Flora other than Bacillus sp. from Bentong Hot Spring in Pahang, Malaysia / Jurnal Sains Kesihatan Malaysia

A study on the isolation and identification of bacterial flora from hot spring was carried out using water from Bentong hot spring in Pahang. In general most of the bacteria identified from the hot springs were from Bacillus sp. This was due to its ability to adapt to high temperature mainly by forming endospore. The objective of this study was to identify bacteria other than Bacillus that were able to survive in hot springs. Three hot spring pools were identified (A,B and C) with temperatures ranging from 36°C to 52°C and pH values between 7.88-8.65. This study has succesfully isolated two species of Gram positive cocci and three species of Gram negative bacilli. The two Gram positives were Streptococcus sp. and Staphylococcus sp. The three species of Gram negative rods were unidentified and were denoted as X, XI and XII.

Effects of Hot Spring Bathing on Pregnancy and Labor-3rd report- / 日本温泉気候物理医学会雑誌

We studied the effect of hot-spring bathing by pregnant women on the Apgar score of their vaginally delivered babies by retrospective investigation.<br>The subjects were classified into four groups: group 1 consisting of 33 pregnant women who took hot-spring baths everyday, group 2 consisting of 166 pregnant women who took plain-water baths with additives everyday, group 3 consisting of 308 pregnant women took plain-water baths without additives everyday, and group 4 consisting of 34 pregnant women who showers everyday.<br>In group 1, the Apgar score was 9 for 27 babies (81.8%) and 8 for six babies (18.2%). In group 2, the Apgar score was 10 for two babies (1.2%), 9 for 125 babies (76.2%), 8 for 37 babies (22.6%), 7 for one baby (0.6%), and 6 for one baby (0.6%). In group 3, the Apgar score was 10 for five babies (1.6%), 9 for 227 babies (73.7%), 8 for 69 babies (22.4%), 7 for four babies (1.3%), 6 for one baby (0.3%), 4 for one baby (0.3%), and 3 for one baby (0.3%). In group 4, the Apgar score was 9 for 30 babies (88.2%) and 8 for four babies (11.8%).<br>No significant correlations were observed between the Apgar score and the groups who bathed in different ways (hot-spring bathing, plain-water bathing, or showers).<br>In conclusion, pregnant women can bathe in hot springs without fear of affecting their babies.

Effects of Hot-Spring Bathing on Pregnancy and Labor-2nd report- / 日本温泉気候物理医学会雑誌

We surveyed 1, 312 pregnant women who were puerperal in-patients of this clinic by retrospective investigation for the effect of hot-spring bathing on the birth weights of babies they delivered.<br>The women were classified into eight groups.<br>Group 1 consisted of 14 pregnant women who took hot-spring baths every day and delivered male babies. Group 2 consisted of 115 pregnant women who took plain-water baths with additives every day and delivered male babies. Group 3 consisted of 139 pregnant women who took plain-water baths without additives every day and delivered male babies. Group 4 consisted of 16 pregnant women who took showers every day and delivered male babies. Group 5 consisted of 17 pregnant women who took hot-spring baths every day and delivered female babies. Group 6 consisted of 133 pregnant women who took plain-water baths with additives every day and delivered female babies. Group 7 consisted of 136 pregnant women who took plain-water baths without additives every day and delivered female babies. Group 8 consisted of 17 pregnant women who took showers every day and delivered female babies.<br>The weights of male babies were 3069.8±357.1g in group 1, 3139.3±396.0g in group 2, 3147.8±382.8g in group 3, and 3037.4±363.7g in group 4. The weights of female babies were 2966.2±337.9g g in group 5, 3050.7±390.6g in group 6, 3087.2±353.1g in group 7, and 3047.9±295.6g in group 8.<br>The weights of male babies were proportional to the duration of bathing. The weights of female babies were also proportional to the duration of bathing.<br>The weights of the placentas of male babies were not proportional to the duration of bathing. The weights of the placentas of female babies were proportional to the duration of bathing.<br>The ratio of the weights of male babies to the weights of the placentas was proportional to the duration of bathing. The ratio of the weights of female babies to the weights of the placentas was not proportional to the duration of bathing.<br>The conclusion: The above results demonstrate that pregnant women may take hot-springs baths without any adverse effect on the baby.

Effects of Hot Spring Bathing on Pregnancy and Labor / 日本温泉気候物理医学会雑誌

We investigated the influences of hot-spring bathing on the incidences of abortion and premature birth using questionnaires returned from 768 puerperal in-patients.<br>These patients were classified into four groups: 24 taking a hot-spring bath daily (group 1), 134 taking a bath with additives (group 2), 178 taking a plain water bath daily (group 3), and 35 taking a shower daily (group 4).<br>The incidence of threatened abortion among the ambulatory patients in each group was 4.2% for group 1, 11.9% for group 2, 9% for group 3, and 2.9% for group 4.<br>The incidence of threatened abortion among the hospitalized patients in each group was 4.2% for group 1, 6.7% for group 2, 4.5% for group 3, and 8.6% for group 4.<br>The incidence of threatened premature birth among the ambulatory patients in each group was 12.5% for group 1, 17.2% for group 2, 15.7% for group 3, and 14.3% for group 4.<br>The incidence of threatened premature birth among the hospitalized patients in each group was 0% for group 1, 7.5% for group 2, 3.4% for group 3, and 2.9% for group 4.<br>The incidence of vaginitis among the patients in each group was 50% for group 1, 43.4% for group 2, 46.6% for group 3, and 44.1% for group 4.<br>The incidence of premature rupture of membrane (PROM) among the patients in each group was 4.2% for group 1, 21.1% for group 2, 12.9% for group 3, and 22.9% for group 4.<br>The incidence of premature birth among the patients in each group was 0% for group 1, 3% for group 2, 2.8% for group 3, and 2.9% for group 4.<br>Among the 42 multiparas experiencing single delivery and being treated for threatened abortion, those who for more than 10 minutes daily showed a significant difference from ambulatory patients being treated for threatened abortion that required hospitalization.<br>Many of the 63 primiparas who did not use a labor accelerating medicine but bathed for more than 10 minutes daily delivered their babies within 1000 minutes.<br>Conclusion<br>The above suggests that pregnant women may bathe in hot-springs without problem but bathing for less than 10 minutes is recommended during early stage of pregnancy.

Effect of Hot Spring Water Drinking on Glucose Metabolism / 日本温泉気候物理医学会雑誌

Drinking of hot spring water containing sulfur or sodium bicarbonate is reported to be beneficial for diabetic patients. In the present study, sulfur spring water and water from simple thermals were chosen to examine their effects on glucose metabolism. Eleven volunteers including eight type 2 diabetic patients and three healthy subjects drank hot spring water from Kawayu Onsen (K-O, acid alum vitriol hydrogen-sulfide springs, pH1.98), and another six patients from Asukano-yu (A-Y, alkaline simple thermals, pH8.9). Oral glucose tolerance test (OGTT) was performed after drinking tap water (200<i>ml</i>) or K-O spring water (5 times diluted) at a week interval (short term effect). Increments in plasma glucose levels were significantly lower (p<0.05) with predrinking of spring water than those with pre-drinking of tap water. To know the long term effect of drinking, patients drank K-O water twice daily for 4 weeks or A-Y water twice daily for a week. Hemoglobin A_1C levels decreased significantly (p<0.05) by drinking K-O water and the levels of 1.5-Anhydro-D-glucitol increased by A-Y water (NS). From these findings, it is suggested that drinking hot spring water from K-O and A-Y is beneficial for diabetic patients if they have no contraindications for drinking these kinds of spring water.

Balneotherapy for psoriasis at Kusatsu. Value of balneotherapy as alternative and complementary medicine / 日本温泉気候物理医学会雑誌

From March 1990 to September 2001, 24 patients with psoriasis (16 males and 8 females, 54±18 years) were admitted to our hospital to receive balneotherapy using Kusatsu hot-spring water. The psoriasis had been refractory to various treatments including steroid ointment therapy over a long period of time. The patients took a 10-minute 40-42°C hot-spring bath followed by application of vitamine D3 ointment 1-2 times daily for 37±19 days. The main components of the hot-spring water are aluminium, sulphates and chlorides, and its pH is 2.0. The skin symptoms of 20 of 24 cases (83%) were improved through the balneotherapy, while those of the remaining 4 cases were not changed. No side effects were observed. The serum levels of uric acid, GOT and GPT which are reported to be increased slightly did not correlate with the skin symptoms. The serum LDH level which is associated with the skin manifestastions in patients with atopic dermatitis also gave no useful information in the treatment of psoriasis. Although the mechanism of the improvement of skin manifestations is not clarified, balneotherapy at Kusatsu can be useful for the treatment of refractory cases of psoriasis as alternative and complementary medicine

Treatment of 100 Cases of Adult-type Atopic Dermatitis with Kusatu Balneotherapy / 日本温泉気候物理医学会雑誌

From June 1990 to October 1998, 100 patients with adult-type atopic dermatitis (59 males and 41 females, 25±8 years) were admitted to our hospital to receive balneotherapy using Kusatsu hot-spring water. The atopic dermatitis in all but 9 cases occurred while the patients were still under 20 and had been refractory to various treatments including steroid ointment therapy over a long period of time. The patients took a 10-minute 40-42°C hot-spring bath followed by immediate application of white petrolatum 1-2 times daily for 75±46 days. The main components of the hot-spring water are aluminium, sulphates and chlorides, and its pH is 2.0. The skin symptoms of 79 of 100 cases (79%) were improved through the balneotherapy and furthermore pruritus was improved in 55 of the 79 cases (70%). The improvement of skin manifestations was supported by a significant decrease in serum LDH levels. In contrast, pruritus was not improved in the remaining 21 cases who showed no changes in skin symptoms and serum LDH levels. Moreover, changes in the number of <i>Staphylococcus aureus</i> on the skin surface were examined before and after balneotherapy. In the 69 cases examined whose skin symptoms were improved, many <i>Staphylococci aureus</i> were detected in 52 of the cases but not in the other 17 cases before starting balneotherapy. They disappeared in 24 cases and decreased in 18 cases of the 52 cases, but were not changed in the remaining 10 cases through the balneotherapy. On the other hand, the number of <i>Staphylococcus aureus</i> on the skin surface was not changed in 11 of the 14 cases examined whose skin symptoms were not improved. Our previous study reported that bactericidal activity against <i>Staphylococcus aureus</i> is expressed by the co-existence of manganese and iodide ions contained in the hot-spring water under an acidic (pH 2.0-3.0) condition. Thus, the mechanisms of the improvement of skin manifestations through the balneotherapy may be explained by considering bactericidal activity of Kusatsu hot-spring water against <i>Staphylococcus aureus</i> inducing acute flares of skin manifestations. Therefore, balneotherapy at Kusatsu can be useful for the treatment of refractory cases of adult-type atopic dermatitis as a suitable method of skin care.