Association between subjective health status and frequency of visits to acupuncture clinic: A cross-sectional study.

OBJECTIVE: There are few studies on the relationship between the frequency of acupuncture use and subjective health status. Therefore, we investigated this relationship using data of a previously performed cross-sectional survey of patients visiting Japanese acupuncture clinics. METHODS: This study used data from a cross-sectional survey conducted in 2011 on patients visiting 180 acupuncture clinics nationwide that were run by members of the alumni association of Meiji University of Integrative Medicine Faculty of Acupuncture and Moxibustion, and did not provide treatment other than acupuncture. We calculated the frequency of visits to acupuncture clinics (< 24 times, 24-47 times, 48-95 times, or ≥ 96 times per year) and the Short Form-36 (SF-36) summary scores (physical, mental, role/social) based on the response of the questionnaire conducted at the time of the survey. Multiple linear regression analysis with multiple imputation was performed with three SF-36 summary scores as the dependent variables, and the frequency of visits to acupuncture clinics as the independent variable. RESULTS: The questionnaire was distributed to 2,379 outpatients of acupuncture clinics, 1,409 of whom met the criteria and were included in the analysis. More frequent visits to acupuncture clinics were associated with lower scores on all three SF-36 summary scores. Compared to those who visited < 24 times a year, those who visited ≥ 96 times a year had unstandardized regression coefficients (95% confidence interval) of -5.6 (-7.8 to -3.3) for the physical, -2.0 (-3.9 to -0.1) for the mental, and -2.9 (-5.4 to -0.4) for the role/social SF-36. CONCLUSIONS: Frequent visits to acupuncture clinics were associated with poor subjective health status, especially physical health.

Salmonella contamination and hazard analysis in a storage facility for feed materials in Japan.

AIMS: To reduce Salmonella contamination of animal feeds, Salmonella contamination in a feed materials storage facility was investigated, and hazard analysis was performed to acquire the basic information necessary to establish a control method for Salmonella. The result is that managers of feed materials storage facilities will be able to introduce the Hazard Analysis and Critical Control Point (HACCP) systems to quickly and efficiently. METHODS AND RESULTS: A total of 472 samples, including deposits, adhesions, environment samples and materials, were investigated. The detection rate in the storage facility was 5.1% (24/472) over the three-year investigation period. The serovars of detected Salmonella were Agona, Anatum, Bron (or Agbeni), Chester, Infantis, Mbandaka, Minnesota, Muenster, Sinstorf, Typhimurium, O18:- and O1,3,19:-. As a result of hazard analysis after confirming the multiplication factors and detection rate, important hazards were judged to be the outdoor equipment and dust collector. CONCLUSIONS: The risk of Salmonella contamination in storage facilities was clarified. Important Salmonella hazards in storage facilities are Salmonella-contaminated feed materials, multiplication factors such as rainwater and dew condensation, and Salmonella-accumulated dust in dust collectors for fine-powder recovery. These results suggest that proper control of these hazards could reduce the risk of Salmonella in storage facilities. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing actual Salmonella contamination conditions and risk assessment using the hazard analysis at a feed materials storage facility in Japan. This study will consider implications for the feed industry because the hazard analysis clarified the sites of Salmonella contamination, the tendency of contamination and the points for implementing control measures in the feed materials storage facility.

Evaluation of Serotyping of Environmental and Clinical Isolates of Legionella pneumophila using MALDI-TOF MS.

Legionella pneumophila (L. pneumophila) is responsible for most Legionnaire's disease cases diagnosed worldwide. The species includes 16 serogroups, but most Legionnaire's disease cases (85.7% in Europe, 87.0% in Japan) are caused by L. pneumophila serogroup 1. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can be used to identify the L. pneumophila serogroup. In this study, we compared three sample preparation methods that are compatible with MALDI-TOF MS: the direct colony transfer method (DCTM), on-target extraction method (OTEM), and in-tube extraction method (ITEM). The aim was to improve the low identification rates for L. pneumophila, and establish and validate a simple, rapid and robust MALDI-TOF MS-based method for routine use in microbiological laboratories for assignment of L. pneumophila isolates to serogroups and identification of reliable peak biomarkers. Using ITEM, 100.0% (29/29) of hot spring water samples and clinical isolates were correctly identified at the species level. Augmented reference spectra correctly identified all 29 strains at the species level and 29 isolates at the serogroup level, displaying sensitivity, specificity and accuracy of 100.0% for serogroup assignment. MALDI-TOF MS is a relatively inexpensive method for assignment of L. pneumophila serogroups that can serve as a first-line tool for rapid prospective typing.

Pilot Study of Acupuncture's Antispasmodic Effect on Upper Gastrointestinal Tract during Endoscopic Submucosal Dissection for Early Gastric Cancer: Controlled Clinical Trial.

A prospective study was conducted in patients with early-stage gastric cancer to determine the efficacy and safety of acupuncture stimulation as an antispasmodic compared with conventional medication during the procedure of endoscopic submucosal dissection (ESD) of the upper gastrointestinal tract. This study was a prospective single blinded quasi-randomized controlled trial. Seventy-three patients who were scheduled to undergo ESD for gastric cancer at Aizu Medical Center between 19 February 2016 and 30 June 2016 were assessed for eligibility for the study. Sixty out of 73 patients were included in the study and assigned into two intervention groups: medication group (MG) and acupuncture group (AG). Ease of the procedure was evaluated using modified NIWA classification (MNC) by endoscopist considering the frequency and amplitude of the upper gastrointestinal peristalsis. For the statistical analysis, Mann-Whitney test was used to compare the differences of MNC values (baseline and end of procedure) between two groups. The difference of MNC found in the AG (-2.00 (-3.0 to -2.0)) was significantly greater than that in the MG (-1.00 (-2.0 to -1.0), p < 0.0001, Mann-Whitney test). We consider that acupuncture to the abdomen could be an alternative antispasmodic method during upper gastrointestinal endoscopic procedure.

[Behavior of Staphylococcus aureus and Staphylococcal Enterotoxins A and Q in Scrambled Eggs].

Staphylococcal food poisoning (SFP) is caused by Staphylococcus aureus, and its typical symptom of vomiting is evoked by staphylococcal enterotoxins (SEs). SEs are classified as classical and new types. SEQ is a new-type enterotoxin predicted to have a high potential risk for SFP. To elucidate the correlation between the number of S. aureus cells and the production of SEs as well as classical and new-type enterotoxins in the food environment, the numbers of S. aureus strain cells carrying sea and seq genes and the production of SEA and SEQ protein were examined under 3 pHs values (pH 6.0, 7.0 and 8.0) and 2 NaCl concentrations (0.5 and 1.0%) conditions. The experiments were performed at 25℃, resembling the setting of scrambled eggs at room temperature after cooking. By 24 hr after incubation, the cell number in the scrambled egg was ≥107/10 g, reaching 109/10 g by 48 hr under all conditions. The productions of both SEA and SEQ were detected in the scrambled egg under all conditions by 48 h. SEQ was detected from 24 hr at all 3 pH values in the egg containing 1.0% NaCl, whereas in the egg containing 0.5% NaCl, it was detected from 24 hr at pH 6.0 and from 48 hr at other pHs. The SEQ production was consistently 100-1,000 times less than that of SEA. These results suggest that the new-type enterotoxin SEQ has the potential to evoke symptoms related to SFP following the consumption of egg products cooked under relative lower pH and water activity.

Effects of acupuncture on nutritional state of patients with stable chronic obstructive pulmonary disease (COPD): re-analysis of COPD acupuncture trial, a randomized controlled trial.

BACKGROUND: There are an increasing number of evidences that chronic obstructive pulmonary disease (COPD) is a systemic illness and that bodyweight loss is its prominent manifestation. We focused on the nutritional outcomes to find out the effectiveness of acupuncture on nutritional state of COPD patients and on their prognosis in our previous interventional study. METHODS: The present study is re-analysis of our previous interventional study, COPD Acupuncture Trial (CAT) published in 2012. Data from CAT was re-analyzed in terms of nutritional status, inflammatory biomarkers, and prognostic index. Nutritional states were evaluated by the measurements of body weight, body composition, and muscle strength, and the nutritional hematological examination results (retinol-binding protein (RBP), prealbumin (PA), transferrin (Tf), and hemoglobin (Hb) in serum), and inflammation biomarkers such as carboxyhemoglobin (COHb), High sensitivity C-reactive protein (Hs-CRP), Tumor Necrosis Factor-alpha (TNF-α), Interleukin 6 (IL-6), and Serum Amyloid A (SAA) were measured. The BODE index was measured in terms of prognosis. These measurements were compared between the real acupuncture group (RAG) and the placebo acupuncture group (PAG). All data are presented as mean (SD) or mean (95% CI). The difference between baseline and final volumes was compared using analysis of covariance (ANCOVA). Moreover, correlations between nutritional hematological examination scores and inflammation biomarker parameters were assessed using Spearman's rank correlation coefficient. RESULTS: After 12 weeks, the change in body weight was significantly greater in the RAG compared with the PAG (mean [SD] difference from baseline: 2.5 [0.4] in RAG vs - 0.5 [1.4] in PAG; mean difference between the groups: 3.00, 95% CI, 2.00 to 4.00 with ANCOVA). Patients in RAG also had improvements in the results of nutritional hematological examination (RBP, PA, Tf, Hb), Inflammation biomarkers (TNF-α, IL-6, SAA, Hs-CRP, COHb) and the BODE index. CONCLUSION: This study demonstrated some clear evidences that acupuncture can be a useful adjunctive therapy to improve nutritional state of COPD patients. TRIAL REGISTRATION: UMIN Clinical Trials Registry ( UMIN000001277 ). Retrospectively registered.

The sensitivity of commercial kits in detecting the genes of pathogenic bacteria in venison.

The expansion of the wild deer population is a major problem for the Japanese farm and forestry industries because their damage to farm products and vegetation results in huge economic loss. To promote game meat consumption, hygiene inspections should be performed to detect main bacterial pathogens before products are shipped. In this study, we aimed to evaluate the ability of commercial test kits to genetically detect EHEC, Salmonella and Listeria monocytogenes in venison. Our results demonstrated that the kits for three pathogens could be useful for venison as well as other domestic meat products. Our comparative study showed that the LAMP kits were more sensitive than the RT-qPCR kits in the detection of all of these pathogens.

Isolation, Identification and Antibacterial Susceptibility of Staphylococcus spp. Associated with the Mobile Phones of University Students.

From May 2014 to February 2015, 319 university students (male, n=173; female n=146) of 18 to 24 years of age who carried mobile phones or computer tablets were selected as subjects. Staphylococcus spp. were detected in 101 of 319 samples (31.7%). In the present study, 11 strains of S. aureus were isolated and identified, not all of which were methicillin-resistant Staphylococcus aureus (MRSA). Overall, 14 species were identified, with 11 strains (10.9%) of S. xylosus being isolated at the highest frequency. Following this were eight strains (7.9%) of S. cohnii and seven strains (6.9%) each of S. capitis and S. haemolyticus. Staphylococcus spp. isolation was performed with bacterial samples obtained from the mobile phones of 22 specific subjects (males, n=12; females, n=10). Staphylococcus spp. isolation was performed on days -1, 7 and 30 of the experiment. Staphylococcus spp. were positively detected one or more times in 12 subjects (54.5%). In one subject (8.3%), all three tests were positive. Furthermore, two tests were positive in three (25.0%). In the eight remaining subjects (66.7%) Staphylococcus spp. were detected only once. For the three abovementioned tests, we investigated the pulsed-field gel electrophoresis (PFGE) patterns of the strains derived from the mobile phone and from the fingers of three subjects in whom the same bacterial species were isolated twice. From the cases with similarities between strains derived from the fingers and the mobile phones and cases, with consistency in the strains derived from the mobile phone at different times, commonality was observed in the strains derived from the fingers and mobile phones along with chronological uniformity in the strains derived from the mobile phones. A total of 101 Staphylococcus spp. strains were isolated from mobile phones. According to drug susceptibility tests, 99 strains (98.0%) were found to have some degree of resistance to drugs (excluding one strain each of S. aureus and S. haemolyticus). Among these, the strain that showed the highest level of drug resistance was one strain (1.0%) of Staphylococcus spp., which showed resistance to nine drugs. The strain that showed the second highest level of drug resistance was one strain (1.0%) of S. caprea, which showed resistance to seven drugs. In this manner, the drug-resistant tendencies of Staphylococcus spp. isolated from mobile phones were observed.

Legionella thermalis sp. nov., isolated from hot spring water in Tokyo, Japan.

Strain L-47(T) of a novel bacterial species belonging to the genus Legionella was isolated from a sample of hot spring water from Tokyo, Japan. The 16S rRNA gene sequences (1477 bp) of this strain (accession number AB899895) had less than 95.0% identity with other Legionella species. The dominant fatty acids of strain L-47(T) were a15:0 (29.6%) and the major ubiquinone was Q-12 (71.1%). It had a guanine-plus-cytosine content of 41.5 mol%. The taxonomic description of Legionella thermalis sp. nov. is proposed to be type strain L-47(T) (JCM 30970(T) = KCTC 42799(T)).

Bacterial contamination in cold water samples obtained from water dispensers.

We carried out a basic study in order to evaluate the bacterial contamination in water dispensers. Water samples were obtained from water dispensers from October 2012 to November 2013, and standard plate counts (at 36˚C, 24 h) of the samples, as well as heterotrophic plate counts (at 25˚C, 7 d), were estimated with the standard methods for the examination of drinking water in Japan. Standard plate counts exceeding the water-quality standard (1.0×10(2) CFU/ml) were observed in 42 of the 140 samples (30.0%), with a maximum detected bacterial count of 2.1×10(5) CFU/ml. The rate of the standard plate counts exceeding the water quality standard tended to be higher when using a one-way type method or water dispensers with natural water. Ralstonia spp. was most commonly isolated, and Pseudomonas aeruginosa was isolated in a few cases. Some opportunistic pathogens were also isolated, suggesting that we should be more concerned about bacterial contamination in cold water supplied from water dispensers.

Antibacterial susceptibility of enterobacteriaceae isolated from raw horsemeat isolated for human consumption (basashi).

Drug susceptibility testing was carried out using 14 antibiotics in order to identify trends in the antibiotic tolerance of 142 strains of Enterobacteriaceae isolated from horsemeat commercially available for raw consumption (basashi). A comparison of the sensitivity to the 14 antibiotics using the 90% MIC (minimum inhibitory concentration) values (MIC90) showed the strongest tolerance to ampicillin (ABPC) at a concentration of > 128 µg/mL, followed by that to fosfomycin (FOM) at a concentration of 128 µg/mL. When the sensitivity to these antibiotics was examined for each individual genus of tested bacteria, Hafnia spp. exhibited relative tolerance to ceftazidime (CAZ) and ceftriaxone (CTRX) at a concentration of 4 µg/mL and 2 µg/mL, respectively, which was high in comparison to that observed for the other strains. Furthermore, Raoultella spp. and Serratia spp. were found to be highly resistant to tetracycline (TC) at a concentration of 128 µg/mL and 64 µg/mL, respectively. Of the 142 strains of test bacteria, 140 (98.6%) demonstrated resistance to ABPC, with the exception of Hafnia alvei and Klebsiella pneumonia. In addition, a total of eight strains (5.6%), seven Serratia marcescens strains and one Raoultella terrigena strain, were found to be resistant to TC. Furthermore, one strain of Citrobacter freundii exhibited resistance to nalidixic acid (NA), while another displayed resistance to ofloxacin (OFLX) (0.7% each), and one strain (0.7%) each of Enterobacter cloacae, Serratia marcescens and Citrobacter youngae demonstrated resistance to fosfomycin (FOM), streptomycin (SM) and kanamycin (KM), respectively. A single strain of C. freundii was found to be resistant to three antibiotics, ABPC, NA and OFLX. Resistance to two antibiotics was confirmed in 11 strains, including seven strains of S. marcescens and one strain of R. terrigena (a total of eight strains) resistant to ABPC and TC, and one strain each of C. youngae, S. marcescens and E. cloacae resistant to ABPC and KM, ABPC and SM, and ABPC and FOM, respectively. In addition, 128 strains were resistant to the single antibiotic of ABPC alone. Of the 140 strains demonstrating antibiotic resistance, 137 (97.9%) retained the conjugative R-plasmid transfer factor, excluding three strains of S. marcescens. All transfer factors were ABPC and retained by a high proportion of the bacterial groups, with one strain (100%) being resistant to three antibiotics, nine (81.8%) of the 11 strains being resistant to two antibiotics, and 127 (99.2%) of the 128 strains being resistant to a single antibiotic. In addition, we examined ESBL productivity in the 140 strains of bacteria demonstrating drug tolerance; however, no strains exhibited this characteristic. Therefore, further observation is required to ascertain trends in antibiotic-tolerant bacteria.

A galactose-binding lectin isolated from Aplysia kurodai (sea hare) eggs inhibits streptolysin-induced hemolysis.

A specific galactose-binding lectin was shown to inhibit the hemolytic effect of streptolysin O (SLO), an exotoxin produced by Streptococcus pyogenes. Commercially available lectins that recognize N-acetyllactosamine (ECA), T-antigen (PNA), and Tn-antigen (ABA) agglutinated rabbit erythrocytes, but had no effect on SLO-induced hemolysis. In contrast, SLO-induced hemolysis was inhibited by AKL, a lectin purified from sea hare (Aplysia kurodai) eggs that recognizes α-galactoside oligosaccharides. This inhibitory effect was blocked by the co-presence of d-galactose, which binds to AKL. A possible explanation for these findings is that cholesterol-enriched microdomains containing glycosphingolipids in the erythrocyte membrane become occupied by tightly stacked lectin molecules, blocking the interaction between cholesterol and SLO that would otherwise result in penetration of the membrane. Growth of S. pyogenes was inhibited by lectins from a marine invertebrate (AKL) and a mushroom (ABA), but was promoted by a plant lectin (ECA). Both these inhibitory and promoting effects were blocked by co-presence of galactose in the culture medium. Our findings demonstrate the importance of glycans and lectins in regulating mechanisms of toxicity, creation of pores in the target cell membrane, and bacterial growth.

Pulsed-Field Gel Electrophoresis (PFGE) pattern analysis and chlorine-resistance of Legionella pneumophila isolated from hot spring water samples.

For a microbial ecological analysis, 20 strains of Legionella pneumophila isolated from both unchlorinated Noyu (unattended natural hot spring) samples and chlorinated hot spring bath water samples collected throughout Japan were subjected to a clustering analysis on the basis of a Pulsed-Field Gel Electrophoresis (PFGE) pattern analysis. The PFGE patterns obtained from 19 strains of L. pneumophila after digestion with SfiI were used to divide the strains into two groups (Groups A and B), although the similarity level was very low among the groups. Group A consisted of 8 strains, and all of these strains were isolated from hot spring bath water samples. Group B consisted of 11 strains, and all but two of these strains were isolated from Noyu samples. The chlorine resistance (99.9% CT values) of these isolates was experimentally confirmed, and we attempted to define the relationship between chlorine resistance and the geno-cluster. The average CT value of group A (8 strains from hot spring bath water) was 0.49 mgï½¥min/l and the average of group B (9 strains from Noyu samples) was 0.29 mgï½¥min/l. No remarkable differences in the CT values for the groups were found. A chlorine-sensitive Noyu strain (0.14 mgï½¥min/l) and a chlorine-resistant strain (0.62 mgï½¥min/l) from hot spring bath water were then compared to identify any differences in their lipid composition. There was no notable difference in the ratio of saturated to unsaturated fatty acids between the chlorine-sensitive and chlorine-resistant bacteria. However, the chlorine-sensitive and chlorine-resistant bacteria demonstrated differences in the relative percentages of cell wall and cell membrane fatty acids.

Structural determination of a nivalenol glucoside and development of an analytical method for the simultaneous determination of nivalenol and deoxynivalenol, and their glucosides, in wheat.

Trichothecene mycotoxins such as nivalenol and deoxynivalenol frequently contaminate foodstuffs. Recently, several trichothecene glucosides have been found in trichothecene-contaminated foods, and information about their chemistry, toxicity, and occurrence is required. In this study, a glucoside of nivalenol was isolated from nivalenol-contaminated wheat and was identified as nivalenol-3-O-ß-D-glucopyranoside. Analytical methods using a multifunctional column or an immunoaffinity column have been developed for the simultaneous determination of nivalenol, nivalenol-3-O-ß-D-glucopyranoside, deoxynivalenol, and deoxynivalenol-3-O-ß-D-glucopyranoside in wheat. The methods were validated in a single laboratory, and recovery from wheat samples spiked at four levels ranged between 86.4 and 103.5% for the immunoaffinity column cleanup. These mycotoxins in contaminated wheat samples were quantitated by the validated method. Nivalenol-3-O-ß-D-glucopyranoside was detected in the nivalenol-contaminated wheat, and the percentage of nivalenol-3-O-ß-D-glucopyranoside to nivalenol ranged from 12 to 27%. This result indicates that the analytical method developed in this study is useful for obtaining data concerning the state and level of food contamination by nivalenol, deoxynivalenol, and their glucosides.

Isolation and identification of Enterobacteriaceae from raw horsemeat intended for human consumption (Basashi).

The status of Enterobacteriaceae contamination was investigated in a total of 131 samples of raw horsemeat (Basashi) intended for human consumption purchased from a general meat shop or by mail-order from October 2012 to December 2013. The bacteria were isolated from 105 of the 112 samples (93.8%). Prominent differences in the isolation rate due to the place of manufacture/sale or by the cut of the meat were not observed. Moreover, in a comparison between domestic (92.6%) and imported (100%) samples, the isolation rate was slightly higher in the imported samples. When Enterobacteriaceae isolated from raw horsemeat was identified, it was highly diverse, with 14 species identified in total. From among these species, Hafnia alvei was the most common, with 33 strains (19.8%),followed by 27 strains (16.2%) of Klebsiella pneumoniae and 26 strains (15.6%) of Enterobacter cloacae, indicating that these three species were dominant. A trend was observed, with the dominant strain differing depending on the place of manufacture/sale or the cut of the meat. H. alvei was isolated at an especially high frequency from imported samples. An investigation was carried out regarding raw horsemeat intended for human consumption from Yamanashi Prefecture and Canada, regularly purchased from one store in Kanagawa Prefecture. Enterobacteriaceae were isolated during five of nine (55.6%） trials, in which the isolated bacteria were H. alvei, K. pneumoniae, etc. Moreover, they were isolated at a very high isolation rate of seven among 10 trials for the Canadian meat, and H. alvei was the most commonly isolated bacteria. Accordingly, when an investigation was carried out regarding the differences in the strain level in the six isolates of H. alvei periodically isolated from raw horsemeat from Canada by the pulsed-field gel electrophoresis (PFGE) pattern using a restriction enzyme, SfiI, there was a possibility that these were the same H-38 strain (November 2013) and H-64 strain (April 2014) as well as the same H-104 strain (July 2014) and H-131 strain (December 2014). As mentioned above, it has been demonstrated that a variety of Enterobacteriaceae were isolated from raw horsemeat (Basashi) intended for human consumption, and at a high frequency. Moreover, based on the fact that the same species or strain was chronologically isolated, the possibility of contamination by the same contamination source at different times was suggested.

Roseomonas tokyonensis sp. nov. isolated from a biofilm sample obtained from a cooling tower in Tokyo, Japan.

Strain K-20(T), a Gram-negative, nonmotile, nonspore-forming and strictly aerobic coccobacillus, which produces a pale pink pigment (R2A agar medium, 30℃, seven days) was isolated from a sample of biofilm obtained from a cooling tower in Tokyo, Japan. A phylogenetic analysis of the 16S rRNA partial gene sequences (1,439 bp) showed that the strain (accession number: AB297501) was related to Roseomonas frigidaquae CW67(T) and Roseomonas stagni HS-69(T) with 97.4% and 96.9% sequence similarity, respectively. Strain K-20(T) formed a distinct cluster with Roseomonas frigidaquae CW67(T) in the phylogenetic tree at a high bootstrap value (93%); however, distance was recognized between the strains. In addition, the DNA-DNA hybridization level between strain K-20(T) and Roseomonas frigidaquae JCM 15073(T) was 33%. The taxonomic data indicate that K-20(T) (=JCM 14634(T) =KCTC 32152(T)) should be classified in the genus Roseomonas as the type strain of a novel species, Roseomonas tokyonensis sp. nov.

Isolation of Legionella species from Noyu (unattended natural hot springs in mountains and fields) samples in Japan.

In order to understand the habitation conditions of the bacteria of the genus Legionella in Noyu (unattended natural hot springs in mountains and fields) in Japan, isolation of Legionella spp. was attempted in the Noyu samples from 11 prefectures nationwide between May and September 2012, and the following results were obtained. Overall, Legionella spp. was isolated from 16 of 43 samples (37.2%). The species was isolated from the Hokkaido region to the Chugoku region but not from the Shikoku region to the Kyushu region. The number of bacteria detected was usually small, less than 5.0 × 10(1) CFU/100 ml, as found in 11 samples (68.8%), while counts of 10(2) or more to 10(3) or less CFU/100 ml were found in two samples (12.5%). Legionella pneumophila was the most commonly found strain, with 19 strains (90.5%) found, and was the dominant species. Regarding the serogrouping, four strains (21.1%) fell under group 1, the most common grouping, followed by three strains (15.8%) in group 3, two strains (10.5%) in group 5, etc. Moreover, the detected bacterial strains other than L. pneumophila included two strains (9.5%) of L. londiniensis. The temperature of the Noyu from which Legionella spp. was isolated was between 33.1°C and 41.5°C with a pH ranging from 5.2 to 8.1. The present report is the first report to clarify the habitation conditions of strains of Legionella spp. isolated from Noyu in Japan.

Characteristics of an orange-pigmented bacterium isolated from a hot spring in Kagoshima, Japan.

In May 2012, strain HNN-6 (=JCM 18566) , a Gram-negative, non-spore-forming, motile and strictly aerobic rod, which produces a pale orange pigment, was isolated from a hot spring water sample obtained in Kagoshima, Japan, by a plating method using R2A medium at 30°C for 7 d. The 16S rRNA gene sequences (1,437bp) of this strain (accession number: AB731137) had a close similarity (99.1%) to Hydrotalea sandarakina AF-51T (JF739858) . Growth occurred at 25-45°C and pH 5.0-8.0, with optimal growth at 40°C and pH 6.0-7.0. Growth did not occur in the presence of ≧2% NaCl. The API 20NE identification system gave positive results for nitrate, aesculin, gelatin, 4-nitrophenyl-ß-D-galactopyranoside, D-glucose, D-mannose, maltose and oxidase (API code number 1472204) . The dominant cellular fatty acids of strain HNN-6 were iso-C15:0 (32.6%) , iso-C17:0 3-OH (24.2%) and iso-C16:0 (8.4%) . The guanine-plus-cytosine (G+C) content of DNA was 36.2 mol%. This article is the first report to describe the characteristics of an orange-pigmented bacterium isolated from a hot spring water sample in Japan.

Combined standard medication and acupuncture for COPD: a case series.

BACKGROUND: Traditional acupuncture has been used in patients with chronic obstructive pulmonary disease (COPD). However, only a few studies have been performed to determine the efficacy of this treatment. OBJECTIVE: To observe changes in the symptoms of COPD during acupuncture treatment in patients with COPD stratified according to the severity of the disease. METHODS: A prospective case series of 26 patients with dyspnoea on exertion due to COPD was followed from October 2004 to October 2008 in the Departments of Respiratory Internal Medicine, Gifu University of Medicine and Meiji University of Integrative Medicine, Japan. All participants received acupuncture treatments once a week for 10 weeks in addition to standard medication therapy. The main outcome measure was the modified Borg dyspnoea scale after the 6 min walk test (6MWT) and the secondary outcome measure was the BODE index. RESULTS: All 26 patients showed significant improvement in the Borg dyspnoea scale after 10 weeks of acupuncture treatment (from 4.02 (2.85) to 1.96 (1.97), mean difference -2.06, 95% CI -3.03 to -1.09, p=0.0002, paired t test). Improvements in the BODE index, 6MWT and oxygen saturation during exercise, which indicates better reduced dyspnoea on exertion and prognosis, were also found. CONCLUSION: The results of this study suggest that acupuncture treatment has clinically useful effects, at least in the short term, in reducing dyspnoea on exercise in patients with COPD, particularly in those more severely affected.