記述言語：日本語   出版者・発行元：一般社団法人 日本感染症学会  

<p>研究目的：改良PCR法を用いて地域健常者の咽頭肺炎球菌保菌率を調べ，さらに増菌培養/PCR法により決定した血清型分布におけるワクチン接種の影響を検討する．</p><p>方法：健常者測定のフィールドを新たに構築し，2014～2020年に地域健常成人449名（重複なし，65才以上361名）を調査した．肺球球菌検出は<i>lytA</i>のPCR検出で，血清型別は増菌培養/PCR法で行った．</p><p>結果：肺炎球菌咽頭保菌率は46.1%で，既報の培養法による成人保菌調査と比べて約10倍高く，保菌率は2017年まで増加傾向を示した．保菌者207名（被験者の46.1%）から188名の血清型を決定し，18C型が66名と最も多く，次いで14型，4型の順で，7価肺炎球菌ワクチン（PCV7）血清型保菌率は67.6%，13価肺炎球菌ワクチン（PCV13）血清型保菌率は73.4%であった．23価肺炎球菌ワクチン（PPSV23）の血清型保菌率は全保菌者中88.9%であり高いカバー率を示した．PPSV23接種を受けていた被験者は193名（65才以上被験者の53.5%），うち保菌率は38.9％で非接種者の保菌率49.4％よりも有意に低かった．</p><p>結語：改良PCR法により地域成人健常者の46.1％に肺炎球菌保菌が検出されたが，保菌株血清型にPPSV23接種による顕著な影響はなく，PPSV23のカバー率は90.8%だった．</p>

DOI： 10.11150/kansenshogakuzasshi.96.82

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記述言語：英語   出版者・発行元：Journal of Hospital Infection  

Background: Antimicrobial resistance in Staphylococcus aureus imposes a high disease burden. Both phenotypic and genotypic monitoring are key to understanding and containing emerging resistant strains. Aim: Phenotypic monitoring of emerging resistance in S. aureus and correlation of priority strain phenotypes with whole-genome sequencing (WGS) findings. Methods: Antimicrobial susceptibility test results of >40,000 isolates from 213 participating hospitals from 2011 to 2019 were exported from the national Japan Nosocomial Infections Surveillance (JANIS) database. Longitudinal and geographic distribution and prevalence of distinct multi-drug resistance phenotypes (‘resistance profiles’) of S. aureus were examined among hospitals and prefectures. We further conducted a genome sequence analysis of strains with specific resistance profiles of concern. Findings: The overall prevalence of meticillin-resistant S. aureus (MRSA) decreased from 40.3% to 35.1% from 2011 to 2019. However, among dozens of S. aureus resistance profiles, only one profile of a type of MRSA, exhibited a statistically significant increase in inpatient frequency, exceeding 10% during the nine years. This MRSA profile showed resistance to oxacillin, erythromycin and levofloxacin. Analysis of WGS results of S. aureus isolates with this phenotype revealed that most belonged to clonal complex 8, and all carried SCCmec IV, typical of community-acquired MRSA. Conclusion: Tracking distinct resistance profiles deepened our understanding of the overall decrease in MRSA and led to recognition of the emergence of a new resistance phenotype. This study provides a model for future epidemiological research on antimicrobial resistance correlating multi-drug resistance phenotypes with selective genome sequencing, which can be applied to other bacterial species.

DOI： 10.1016/j.jhin.2022.02.011

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記述言語：英語  

DOI： 10.1016/j.jgar.2021.12.029

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Carbapenem-resistant Enterobacterales (CRE) represent a serious threat to public health due to the lack of treatment and high mortality. The rate of antimicrobial resistance of Enterobacterales isolates to major antimicrobials, including carbapenems, is much higher in Vietnam than in Western countries, but the reasons remain unknown due to the lack of genomic epidemiology research. A previous study suggested that carbapenem resistance genes, such as the carbapenemase gene blaNDM, spread via plasmids among Enterobacterales in Vietnam. In this study, we characterized blaNDM-carrying plasmids in Enterobacterales isolated in Vietnam, and identified several possible cases of horizontal transfer of plasmids both within and among species of bacteria. Twenty-five carbapenem-nonsusceptible isolates from a medical institution in Hanoi were sequenced on Illumina short-read sequencers, and 13 blaNDM-positive isolates, including isolates of Klebsiella pneumoniae, Escherichia coli, Citrobacter freundii, Morganella morganii, and Proteus mirabilis, were further sequenced on an Oxford Nanopore Technologies long-read sequencer to obtain complete plasmid sequences. Almost identical 73 kb IncFII(pSE11)::IncN hybrid plasmids carrying blaNDM-1 were found in a P. mirabilis isolate and an M. morganii isolate. A 112 kb IncFII(pRSB107)::IncN hybrid plasmid carrying blaNDM-1 in an E. coli isolate had partially identical sequences with a 39 kb IncR plasmid carrying blaNDM-1 and an 88 kb IncFII (pHN7A8)::IncN hybrid plasmid in a C. freundii isolate. 148–149 kb IncFIA(Hl1)::IncA/C2 plasmids and 75–76 kb IncFII(Yp) plasmids, both carrying blaNDM-1 were shared among three sequence type 11 (ST11) isolates and three ST395 isolates of K. pneumoniae, respectively. Most of the plasmids co-carried genes conferring resistance to clinically relevant antimicrobials, including third-generation cephalosporins, aminoglycosides, and fluoroquinolones, in addition to blaNDM-1. These results provide insight into the genetic basis of CRE in Vietnam, and could help control nosocomial infections.

DOI： 10.1371/journal.pone.0231119

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

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記述言語：日本語   出版者・発行元：(一社)日本消化器内視鏡学会  

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記述言語：英語   出版者・発行元：株式会社医学書院  

DOI： 10.11477/mf.1543208043

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記述言語：英語   出版者・発行元：株式会社医学書院  

DOI： 10.11477/mf.1401208754

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：日本衛生動物学会  

<p>In Japan, fourteen bites by the red back spider, <i>Latrodectus hasseltii</i>, were reported in four hospitals between 2011 and 2013 in a survey of sentinel hospitals. The distribution of the spider and the areas in which patients were bitten by the spider both expanded geographically each year. Although fatalities or severely ill patients are yet to be reported in Japan, stockpiles of the antivenom and communication with the public and medical professionals should be considered.</p>

DOI： 10.7601/mez.67.219

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記述言語：英語   出版者・発行元：Lecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics)  

Students build cognitive models for solving a cryptarithmetic task in a learning environment that enables them to formally describe various types of procedural knowledge in a group learning setting in which each student is allowed to refer to the procedural rules described by the other group members. Experimental evaluation showed that: (1) three-quarters of participants successfully constructed valid models with the system, and (2) participants learned to describe procedural knowledge more precisely not only for the training task (cryptarithmetic task) but also for a transfer task (bug identification for a multi-column subtraction problem).

DOI： 10.1007/978-3-319-39583-8_1

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：公益社団法人 日本薬学会  

国，地域，病院，または病棟ごとに分離された細菌の薬剤感受性試験のデータを集計し，それぞれの菌種で各種抗菌薬について耐性，中間，感性の菌がどれくらいの割合であるのかを図や表にしたもの．薬剤耐性菌対策を検討していくための重要な基本情報となる．また，臨床現場における治療薬のガイドラインを作成する際には，重要な参考情報となる．さらに，病院や病棟ごとでデータの経時的な変化を見たり，そのデータを地域や国全体のデータと比較することで，感染制御がうまくできているかどうかを評価することにも用いられる．

DOI： 10.14894/faruawpsj.51.4_342_2

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：公益社団法人 日本薬学会  

20世紀に入ってからペニシリンをはじめとする様々な抗菌薬が開発され，人類にとって大きな脅威である感染症から多くの人が救われてきた．しかし一方で，新たな抗菌薬が開発されると間もなく，その薬剤に耐性を獲得した細菌が出現し，薬剤の普及とともに拡散してきた．これまでに耐性が出現しなかった抗菌薬はなく，これは今後も続くと考えられる．近年，薬剤耐性菌は国境を越えて拡散し，世界的な問題になっている．菌種や薬剤によっては，耐性菌の割合が数年で大きく増加しているものもある．WHOは2013年に薬剤耐性菌に関する諮問グループを組織し，2014年に薬剤耐性菌に関する報告書を出した．WHOは，この中で今後取り組むべき課題の1つとして，薬剤耐性菌のグローバルサーベイランスの強化を挙げている．<br>日本においては，薬剤耐性菌のサーベイランスとして厚生労働省院内感染対策サーベイランス（Japan Nosocomial Infection Surveillance：JANIS）事業がある．本稿では，JANISの2013年の公開情報2）のデータを用いて，主要な各種耐性菌の国内での状況を紹介する．さらに，近年特に問題になっているカルバペネム耐性の腸内細菌科細菌について，日本でよく検出される耐性遺伝子や外国の状況との比較なども説明する．また，JANISが各参加医療機関個別に感染対策に活用していただくことを目的に返している報告書についても触れる．

DOI： 10.14894/faruawpsj.51.4_324

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：一般社団法人 日本医療薬学会  

DOI： 10.20825/amjsphcs.24.0_421_6

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Genome Announcements  

Pseudomonas alcaligenes, a Gram-negative aerobic bacterium, is a rare opportunistic human pathogen. Here, we report the whole-genome sequence of P. alcaligenes strain MRY13-0052, which was isolated from a bloodstream infection in a medical institution in Japan and is resistant to antimicrobial agents, including broad-spectrum cephalosporins and monobactams.

DOI： 10.1128/genomeA.e00919-13

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Blackwell Publishing Asia Pty.  

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記述言語：英語   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

DOI： 10.7883/yoken.63.113

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記述言語：日本語   掲載種別：研究論文（学術雑誌）  

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Nippon rinsho. Japanese journal of clinical medicine  

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記述言語：英語  

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記述言語：英語  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Center for Academic Pub. Japan  

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/S0140-6736(03)14959-8

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1128/JCM.41.12.5407-5413.2003

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1128/AAC.47.9.2981-2983.2003

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Molecular Microbiology  

Helicobacter pylori infection induces apoptosis in gastric epithelial cells. Here, we report a novel apoptosis-inducing protein that functions as a leading factor in H. pylori-mediated apoptosis induction. We purified the protein from H. pylori by separating fractions that showed apoptosis-inducing activity. This protein induced apoptosis of AGS cells in a dose-dependent manner. The purified protein consisted of two protein fragments with molecular masses of about 40 and 22 kDa, which combined to constitute a single complex in their natural form. N-terminal sequencing indicated that both these protein fragments were encoded by the HP1118 gene. The purified protein exhibited γ-glutamyl transpeptidase activity, the inhibition of which by 6-diazo-5-oxo-L-norleucine resulted in a complete loss of apoptosis-inducing activity. To the best of our knowledge, the apoptosis-inducing function is a newly identified physiological role for bacterial γ-glutamyl transpeptidase. The apoptosis-inducing activity of the isogenic mutant γ-glutamyl transpeptidase-deficient strain was significantly lower compared with that of the parent strain, demonstrating that γ-glutamyl transpeptidase plays a significant role in H. pylori-mediated apoptosis. Our findings provide new insights into H. pylori pathogenicity and reveal a novel aspect of the bacterial γ-glutamyl transpeptidase function.

DOI： 10.1046/j.1365-2958.2003.03305.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1128/AAC.46.8.2427-2434.2002

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Emerging Infectious Diseases  

Multidrug resistance in gram-positive bacteria has become common worldwide. In Japan until recently, gram-negative bacteria such as Pseudomonas aeruginosa, Klebsiella pneumoniae, and Serratia marcescens were controlled by carbapenems, fluoroquinolones, and aminoglycosides. However, several of these microorganisms have recently developed resistance against many antimicrobial drugs.

DOI： 10.3201/eid0606.000604

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

Recently, it was reported that amoxicillin-clavulanate has slightly higher activity than amoxicillin against Helicobacter pylori. In this study, we evaluated the in-vitro antibacterial activity of β-lactamase inhibitors against H. pylori. We investigated the susceptibility of 30 H. pylori strains to β-lactamase inhibitors, including clavulanate, sulbactam, and tazobactam. In short-term bactericidal studies, a clinical isolate of H. pylori NU27 was exposed to 1 x minimum inhibitory concentration (MIC) of the β-lactamase inhibitors, amoxicillin, clarithromycin, and amoxicillin-clavulanate for 3 and 6h. The MICs90 for these β-lactamase inhibitors were 2, 4, and 2 mg/l, respectively. The short-term bactericidal studies showed that these β- lactamase inhibitors decreased viable counts of H. pylori during 6-h exposure at 1 x MIC. Our results suggest that β-lactamase inhibitors have in-vitro antibacterial activity against H. pylori. Amoxicillin and clavulanate used in combination resulted in increased antibacterial activity.

DOI： 10.1007/s101560050036

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：FEMS Microbiology Letters  

Escherichia coli chromosome encodes several multidrug transporters. Despite their protective function against antibacterial agents, the specific physiological actions of these transporters are not fully understood. E. coli produces indole, a metabolite of tryptophan, under physiological conditions. Defined inactivation of the acrEF gene, the product of which is known as an energy-dependent multiple drug efflux pump, decreased indole excretion while reintroduction of the acrEF gene restored it. A ΔacrEF mutant accumulated more intracellular indole than the parent. This mutant was more susceptible to the growth-inhibitory effect of indole than the parent. These results indicate that the AcrEF system plays a significant role in indole efflux. Copyright (C) 1999 Federation of European Microbiological Societies.

DOI： 10.1016/S0378-1097(99)00433-4

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：財団法人 学会誌刊行センター  

We examined whether anti-Lewis x (Le^x) and y (Le^y) autoantibodies affect the pathogenesis of <i>Helicobacter pylori</i>-induced peptic ulcers. Of 11 patients with peptic ulcers, 10 patients had both anti-Le^x and -Le^y immunoglobulin G (IgG) antibodies, and 1 patient had only anti-Le^x antibody. After successful eradication, we measured the serum titer of anti-Le^x and -Le^y antibodies. Six patients had a reduction of the titers of anti-Le^x and/or -Le^y antibodies, whereas no notable changes were detected in 5 patients in the follow-up. This result suggests that anti-Le^x and -Le^y autoantibodies had no critical role in the development of <i>H. pylori</i>-induced peptic ulcer.

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：FEMS Microbiology Letters  

Escherichia coli K-12 WaaR is a non-processive α-1,2 glucosyltransferase, involved in the synthesis of the R-core of lipopolysaccharide. WaaR possesses the four conserved structural regions I, II, III and IV, each presumably involved in the mechanistic function in catalysis. Regions I and III contain the pair of strictly conserved Asp residues. Asp-129, 131 (region I) and 215, 217 (region III) of WaaR were individually converted to Asn by the site-directed mutagenesis of the waaR gene. All mutated enzymes were inactive, supporting the model for an α-glycosyl transfer reaction where the pair of strictly conserved aspartic acid residues in regions I and III play a critical role in the catalytic function. Copyright (C) 1999 Federation of European Microbiological Societies.

DOI： 10.1016/S0378-1097(99)00127-5

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：財団法人 学会誌刊行センター  

A series of new synthetic ligand compounds which chelate divalent cations was examined for the antibacterial activities of the compounds. Only 2 of 14 synthetic chelators, 9-<i>trans</i>-anthryl-1, 4, 8, 11-tetraazatetradecane (No. 6) and bis(2-pyridyl)methylamine (No. 13) showed antibacterial activities, whereas none of the diamines, hydrophilic triamines nor tetramines showed antibacterial activities. Chelators No. 6 and No. 13 inhibited the growth of both Gram-negative and -positive bacteria at doses of 25-200&mu;g/ml, comparable to those of common antibiotics such as polymixin B, fosfomycin and macrolides. Ethylenediaminetetraacetate (EDTA) potentiated these antibacterial activities, whereas an inhibitory effect of Mg²⁺ on the MICs of these chelators was observed. Moreover, these chelators enhanced the leakage of periplasmic &beta;-lactamase. It is therefore suggested that chelators No. 6 and No. 13 disrupt both the membranes and cytoplasmic functions of bacteria, resulting in cell death.

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Antimicrobial Agents and Chemotherapy  

We evaluated the susceptibilities of 129 Shiga-like toxin-producing Escherichia coli (STEC) isolates to various antibiotics. The numbers of isolates for which MICs were high (≥128 μg/ml) were as follows: 5 for fosfomycin, 14 for ampicillin, 1 for cefaclor, 6 for kanamycin, 22 for tetracycline, and 2 for doxycycline. For two isolates of STEC O26 MICs of fosfomycin were high (1,024 and 512 μg/ml, respectively). Conjugation experiments and glutathione S-transferase assays suggested that the fosfomycin resistance in these isolates was determined not by a plasmid but chromosomally. The amount of active intracellular fosfomycin in these STEC isolates was 100- to 200-fold less than that in E. coli C600 harboring pREFTT47B408 in the presence of either L-α-glycerophosphate or glucose-6- phosphate. Cloning, sequencing, and Northern blot analysis demonstrated that the transcriptional level of the murA gene encoding UDP-N-acetylglucosamine enolpyruvoyl transferase in these isolates was greater than that in E. coli C600. Our results suggest that the fosfomycin resistance in these STEC isolates is due to concurrent effects of alteration of the glpT and/or uhp transport systems and of the enhanced transcription of the murA gene.

DOI： 10.1128/aac.43.4.789

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：財団法人 学会誌刊行センター  

The growth and emetic toxin (cereulide) production of <i>Bacillus cereus</i> strains in defined culture media were studied. We found that a fully synthetic medium (CADM) allowed the production of emetic toxin and the addition of glucose enhanced it. By subtracting each amino acid from CADM, we found that only three amino acids, valine, leucine and threonine, were essential for growth and toxin production by <i>B. cereus</i>. The addition of high levels (50mM) of leucine, isoleucine and glutamlc acid decreased the toxin production. Other amino acids had no effect at this concentration.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Center For Academic Publications Japan  

A series of new synthetic ligand compounds which chelate divalent cations was examined for the antibacterial activities of the compounds. Only 2 of 14 synthetic chelators, 9-<i>trans</i>-anthryl-1, 4, 8, 11-tetraazatetradecane (No. 6) and bis(2-pyridyl)methylamine (No. 13) showed antibacterial activities, whereas none of the diamines, hydrophilic triamines nor tetramines showed antibacterial activities. Chelators No. 6 and No. 13 inhibited the growth of both Gram-negative and -positive bacteria at doses of 25-200μg/ml, comparable to those of common antibiotics such as polymixin B, fosfomycin and macrolides. Ethylenediaminetetraacetate (EDTA) potentiated these antibacterial activities, whereas an inhibitory effect of Mg²⁺ on the MICs of these chelators was observed. Moreover, these chelators enhanced the leakage of periplasmic β-lactamase. It is therefore suggested that chelators No. 6 and No. 13 disrupt both the membranes and cytoplasmic functions of bacteria, resulting in cell death.

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology and Immunology  

We examined whether anti-Lewis x (Le(x)) and y (Le(y)) autoantibodies affect the pathogenesis of Helicobacter pylori-induced peptic ulcers. Of 11 patients with peptic ulcers, 10 patients had both anti-Le(x) and -Le(y) immunoglobulin G (IgG) antibodies, and 1 patient had only anti-Le(x) antibody. After successful eradication, we measured the serum titer of anti- Le(x) and -Le(y) antibodies. Six patients had a reduction of the titers of anti-Le(x) and/or -Le(y) antibodies, whereas no notable changes were detected in 5 patients in the follow-up. This result suggests that anti-Le(x) and - Le(y) autoantibodies had no critical role in the development of H. pylori- induced peptic ulcer.

DOI： 10.1111/j.1348-0421.1999.tb02423.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Center For Academic Publications Japan  

We examined whether anti-Lewis x (Le^x) and y (Le^y) autoantibodies affect the pathogenesis of <i>Helicobacter pylori</i>-induced peptic ulcers. Of 11 patients with peptic ulcers, 10 patients had both anti-Le^x and -Le^y immunoglobulin G (IgG) antibodies, and 1 patient had only anti-Le^x antibody. After successful eradication, we measured the serum titer of anti-Le^x and -Le^y antibodies. Six patients had a reduction of the titers of anti-Le^x and/or -Le^y antibodies, whereas no notable changes were detected in 5 patients in the follow-up. This result suggests that anti-Le^x and -Le^y autoantibodies had no critical role in the development of <i>H. pylori</i>-induced peptic ulcer.

CiNii Research

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology and Immunology  

A series of new synthetic ligand compounds which chelate divalent cations was examined for the antibacterial activities of the compounds. Only 2 of 14 synthetic chelators, 9-trans-anthryl-1, 4, 8, 11-tetraazatetradecane (No. 6) and bis(2-pyridyl)methylamine (No. 13) showed antibacterial activities, whereas none of the diamines, hydrophilic triamines nor tetramines showed antibacterial activities. Chelators No. 6 and No. 13 inhibited the growth of both Gram-negative and -positive bacteria at doses of 25-200 μg/ml, comparable to those of common antibiotics such as polymixin B, fosfomycin and macrolides. Ethylenediaminetetraacetate (EDTA) potentiated these antibacterial activities, whereas an inhibitory effect of Mg2+ on the MICs of these chelators was observed. Moreover, these chelators enhanced the leakage of periplasmic β-lactamase. It is therefore suggested that chelators No. 6 and No. 13 disrupt both the membranes and cytoplasmic functions of bacteria, resulting in cell death.

DOI： 10.1111/j.1348-0421.1999.tb02410.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Center For Academic Publications Japan  

The growth and emetic toxin (cereulide) production of <i>Bacillus cereus</i> strains in defined culture media were studied. We found that a fully synthetic medium (CADM) allowed the production of emetic toxin and the addition of glucose enhanced it. By subtracting each amino acid from CADM, we found that only three amino acids, valine, leucine and threonine, were essential for growth and toxin production by <i>B. cereus</i>. The addition of high levels (50mM) of leucine, isoleucine and glutamlc acid decreased the toxin production. Other amino acids had no effect at this concentration.

CiNii Research

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology and Immunology  

The growth and emetic toxin (cereulide) production of Bacillus cereus strains in defined culture media were studied. We found that a fully synthetic medium (CADM) allowed the production of emetic toxin and the addition of glucose enhanced it. By subtracting each amino acid from CADM, we found that only three amino acids, valine, leucine and threonine, were essential for growth and toxin production by B. cereus. The addition of high levels (50 mM) of leucine, isoleucine and glutamic acid decreased the toxin production. Other amino acids had no effect at this concentration.

DOI： 10.1111/j.1348-0421.1999.tb02367.x

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Infection and Immunity  

Acute respiration distress syndrome (ARDS) is a typical complication in toxic shock-like syndrome (TSLS) caused by Streptococcus pyogenes. An isolated perfused rat lung model was used to identify the causative agent of ARDS in TSLS in this study. Some crude preparations separated from the culture supernatants of S. pyogenes isolates caused rapid increases in the weight of perfused lungs, indicating vascular permeabilization. Six samples from M type 1 and 3 isolates from TSLS and pharyngitis patients showed strong permeabilization activity, whereas preparations from isolates of other M types (although the number of isolates examined was limited) were negative. The active substance was purified to a single band by sodium dodecyl sulfate- polyacrylamide gel electrophoresis using various columns, and the N-terminal amino acid sequence was determined. The resultant sequence of eight amino acids was identical to SpeF (mitogenic factor). Moreover, the vascular permeabilization activity of the purified band was abolished with anti-SpeF antiserum prepared by immunizing with the purified SpeF. This activity was also neutralized by the antiserum prepared by immunizing with a synthetic peptide derived from the published SpeF sequence. These results suggested that streptococcal SpeF is a major cause of permeabilization of lung blood vessels and sufficient for the pathogenesis of ARDS under the conditions of TSLS caused by S. pyogenes.

DOI： 10.1128/iai.67.9.4307-4311.1999

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：財団法人 学会誌刊行センター  

We undertook an epidemiologic study for the sensitivity of both Shiga-like toxin (Slt)-producing <i>Escherichia coli</i> (STEC) O157 and non-STEC O157 strains isolated from different patients with diarrhea to hydrochloric acid (HCl) and organic acids such as acetate, propionate, butyrate and lactate, and other pathogenic factors. The <i>E. coli</i> O157 isolates examined showed a wide variety of organic-acid susceptibility patterns. <i>E. coli</i> O157 isolates resistant to HCl or acetate were found more frequently than those resistant to other organic acids. These isolates also showed diverse pathogenicity patterns for the presence of the virulence genes, antibiotic susceptibility and plasmid profile.

CiNii Research

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Bacteriology  

Escherichia coli K-12 WaaO (formerly known as RfaI) is a nonprocessive α-1,3 glucosyltransferase, involved in the synthesis of the R core of lipopolysaccharide. By comparing the amino acid sequence of WaaO with those of 11 homologous α-glycosyltransferases, four strictly conserved regions, I, II, III, and IV, were identified. Since functionally related transferases are predicted to have a similar architecture in the catalytic sites, it is assumed that these four regions are directly involved in the formation of α- glycosidic linkage from α-linked nucleotide diphospho-sugar donor. Hydrophobic cluster analysis revealed a conserved domain at the N termini of these α-glycosyltransferases. This domain was similar to that previously reported for β-glycosyltransferases. Thus, this domain is likely to be involved in the formation of β-glycosidic linkage between the donor sugar and the enzyme at the first step of the reaction. Site-directed mutagenesis analysis of E. coli K-12 WaaO revealed four critical amino acid residues.

DOI： 10.1128/jb.180.20.5313-5318.1998

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Center For Academic Publications Japan  

We undertook an epidemiologic study for the sensitivity of both Shiga-like toxin (Slt)-producing <i>Escherichia coli</i> (STEC) O157 and non-STEC O157 strains isolated from different patients with diarrhea to hydrochloric acid (HCl) and organic acids such as acetate, propionate, butyrate and lactate, and other pathogenic factors. The <i>E. coli</i> O157 isolates examined showed a wide variety of organic-acid susceptibility patterns. <i>E. coli</i> O157 isolates resistant to HCl or acetate were found more frequently than those resistant to other organic acids. These isolates also showed diverse pathogenicity patterns for the presence of the virulence genes, antibiotic susceptibility and plasmid profile.

CiNii Research

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Antimicrobial Agents and Chemotherapy  

Escherichia coli HKY56, which demonstrated resistance to various β- lactams except carbapenems, was isolated from the throat swab of an inpatient in 1994. Conjugal transfer of cephem resistance from HKY56 to E. coli CSH2 was not successful. Three cefotaxime-resistant E. coli clones harboring plasmid pMRE001, pMRE002, or pMRE003, each of which carried a 3.4-, 5.8-, or 6.2-kb EcoRI fragment insert, respectively, were obtained from HKY56. Although restriction analysis suggested their different origins, these clones showed similar profiles of resistance to various β-lactams. The sequence of 10 amino acid residues at the N terminus of β-lactamase purified from E. coli HB101(pMRE001) was identical to that of Toho-1. This Toho-1-like β- lactamase-1 (TLB-1) was able to hydrolyze cefoperazone and cefotaxime efficiently, but it failed to hydrolyze cephamycins. A Toho-1-specific DNA probe was hybridized with three distinct EcoRI fragments derived from the chromosomal DNA of strain HKY56, and these fragments corresponded to DNA inserts carried by pMRE001, pMRE002, and pMRE003, respectively. PCR and Southern hybridization analysis suggested that all six cephem-resistant E. coli strains, strains HKY273, HKY285, HKY288, HKY305, HKY316, and HKY335, which were isolated in 1996 at the same hospital where strain HKY56 had been isolated, also possessed multiple Toho-1-like β-lactamase (TLB) genes, and the hybridization patterns obtained with the Toho-1-specific probe were quite similar among these six isolates. The DNA fingerprinting patterns observed by pulsed-field gel electrophoresis revealed that among the E. coli isolates tested, all isolates except HKY56 possessed a similar genetic background. These findings suggested that E. coli strains that carry chromosomally multiplied TLB genes may have been proliferating and transmitted among patients in the same hospital.

DOI： 10.1128/aac.41.12.2606

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：財団法人 学会誌刊行センター  

RobA is a member of the XylS/AraC subfamily of DNA binding proteins, and when overexpressed, it induces multiple antibiotic resistance in <i>Escherichia coli</i>. In this study, we introduced a multicopy <i>robA</i> plasmid (pMEP1) and its derivative into OmpF mutants and an AcrAB-deficient mutant. We found that a decrease in susceptibility to multiple antibiotics in these OmpF mutants when pMEP1 was introduced did not depend on OmpF porin expression. Interestingly, a <i>&Delta;ompF</i> mutant (TK007) became more sensitive when pMEP1 was introduced. Moreover, no effect of RobA on the induction of multiple antibiotic resistance in an <i>acrA1</i>^- mutant was observed. Therefore, we conclude that the multiple antibiotic resistance induced by the overexpression of RobA largely depends on the activation of the AcrAB efflux, as well as the activation of <i>micF</i>.

CiNii Research

記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology and Immunology  

RobA is a member of the XyIS/AraC subfamily of DNA binding proteins, and when overexpressed, it induces multiple antibiotic resistance in Escherichia coli. In this study, we introduced a multicopy robA plasmid (pMEP1) and its derivative into OmpF mutants and an AcrAB-deficient mutant. We found that a decrease in susceptibility to multiple antibiotics in these OmpF mutants when pMEP1 was introduced did not depend on OmpF porin expression. Interestingly, a ΔompF mutant (TK007) became more sensitive when pMEP1 was introduced. Moreover, no effect of RobA on the induction of multiple antibiotic resistance in an acrA1- mutant was observed. Therefore, we conclude that the multiple antibiotic resistance induced by the overexpression of RobA largely depends on the activation of the AcrAB efflux, as well as the activation of micF.

DOI： 10.1111/j.1348-0421.1997.tb01913.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：STAR Protocols  

Infection with Helicobacter suis, which causes many cases of gastric disease, is not reliably diagnosed. Here, we present a protocol for detecting H. suis infection. We describe steps for collecting gastric biopsies and sera from patients, preparing DNA for PCR, and targeting the H. suis-specific gene. We then define procedures for inoculating biopsies onto primary agar plates and transferring colonies to secondary agar plates. Finally, we detail whole-genome sequencing of bacteria and assess H. suis infection in sera with ELISA. For complete details on the use and execution of these protocols, please refer to Matsui et al.1

DOI： 10.1016/j.xpro.2023.102556

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記述言語：日本語   出版者・発行元：(株)ライフ・サイエンス  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Medical Microbiology  

Introduction. Carbapenemase-producing Enterobacteriaceae (CPE) have emerged as a global threat to public health and clinical practice. Hypothesis/Gap Statement. In Thailand, reports describing CPEs carrying blaNDM and blaOXA-48-like genes have been increasing recently; however, data on detailed plasmid analysis and temporal shift of sequence type and carbapenemase type are limited. Aim. In this study, we analysed whole-genome sequencing (WGS) data of clinically isolated carbapenemase-producing Klebsiella pneumoniae (CPKP) to reveal the molecular epidemiology of CPKP in a tertiary-care hospital in Bangkok, Thailand. Methodology. Seventy-seven non-duplicated CPKP isolates collected during 2013–2016 were examined for their drug-resistance genes, sequence types and phylogenetic relationships. Results. All the tested isolates possessed carbapenemase gene(s), and the major type of carbapenemase gene in 2014–2015 was blaNDM-1, whereas isolates in 2016 harboured more blaOXA-232 than blaNDM-1. Other carbapenemase gene variants, such as blaNDM-4, blaNDM-5, blaOXA-48, blaOXA-181 and blaIMP-14 were detected in some CPKP isolates. Furthermore, this study revealed that CPKP co-harbouring two genes, blaNDM-1 and blaOXA-232 or blaOXA-181, emerged during this period. Notably, such isolates co-carrying the two carbapenemase genes emerged in three different sequence types, even in a single hospital, and then spread clonally. The WGS of CPKP revealed a temporal shift of the predominant carbapenemase genes from blaNDM-1 to blaOXA-232 along with a variation in other carbapenemase gene types within a span of 4 years. Conclusion. Our findings suggest that a substantial change in CPE types occurred in Thailand and potentially in Southeast Asian countries.

DOI： 10.1099/jmm.0.001711

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Emerging Infectious Diseases  

We report the isolation of Helicobacter ailurogastricus, a Helicobacter species that infects cats and dogs, from a person with multiple refractory gastric ulcers. In addition to H. suis, which infects pigs, Helicobacter species that infect cats and dogs should be considered as potential gastric pathogens in humans.

DOI： 10.3201/eid2904.221807

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Medical Entomology  

Bartonella quintana is a gram-negative bacterium causing trench fever, an illness historically acquired by soldiers during World War I. More recently, outbreaks of trench fever have been reported in those experiencing homelessness in the United States, France, Russia, andTokyo, as well as in children in Nepal and persons in Ethiopia. Reports of B. quintana infection outside ofTokyo are rare in Japan.The aim of this study was to examine body lice and blood obtained from people staying in shelters in Osaka (2009-2010) for B. quintana via polymerase chain reaction and enzyme-linked immunosorbent assays. Day laborers were defined as homeless individuals and shelter residents in this study. We detected genes of B. quintana in body lice by PCR and antibodies against B. quintana. The positive rate of B. quintana genes was 6/10 (60%) in body lice and the seroprevalence (IgG) of B. quintana was 4/10 (40%).This demonstrates that trench fever was endemic in people staying in shelters in Osaka in 2009-2010.

DOI： 10.1093/jme/tjad001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Background Surgical site infections (SSIs) are among the most common healthcare-associated infections. Laparoscopy is increasingly being used in various surgical procedures. However, no study has examined the association between the proportion of laparoscopic procedures and the incidence of SSIs in digestive surgery using nationwide surveillance data. Methods We retrospectively investigated national SSI surveillance data from the Japan Nosocomial Infections Surveillance between 2009 and 2019. The annual trend of the SSI rate and the proportion of laparoscopic procedures were assessed, focusing on five major digestive surgeries. This was based on data from 109,544 (appendix surgery), 206,459 (gallbladder surgery), 60,225 (small bowel surgery), 363,677 (colon surgery), and 134,695 (rectal surgery) procedures. The effect of a 10% increase in the proportion of laparoscopic procedures on the reduction of the SSI rate was estimated using mixed-effect logistic regression. Findings The average SSI rate of the five digestive surgeries decreased from 11.8% in 2009 to 8.1% in 2019. The proportion of laparoscopic procedures in each of the five digestive surgeries increased continuously (p<0.001). The SSI rate for laparoscopic procedures was always lower than that for open procedures. The results were consistent between all and core hospitals participating in the surveillance. The odds ratios of the 10% increase in the proportion of laparoscopic procedures for five digestive surgeries were always <0.950 (p<0.001). Conclusion An increase in the proportion of laparoscopic procedures was associated with a reduction in the SSI rate in digestive surgeries.

DOI： 10.1371/journal.pone.0281838

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記述言語：英語   出版者・発行元：日本細菌学会  

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記述言語：英語   出版者・発行元：日本細菌学会  

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記述言語：英語   出版者・発行元：日本細菌学会  

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記述言語：英語   出版者・発行元：株式会社医学書院  

DOI： 10.11477/mf.1403203095

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

近年、H.pylori(H.p.)感染者の多くが除菌を受け、また衛生環境が良くなったことでH.p.感染者は減少したが、一方でH.p.以外のHelicobacter(NHPH)による胃疾患が相対的に増加している。胃型NHPHには、H.suis、H.felis、H.heilmannii、H.salomonisなどが存在する。1)H.p.感染の胃MALTリンパ腫、2)H.p.未感染の胃MALTリンパ腫、3)胃MALTリンパ腫とNHPH感染、4)NHPH感染胃MALTリンパ腫の内視鏡像、5)H.p.未感染胃MALTリンパ腫に対する除菌効果、6)NHPH感染胃MALTリンパ腫に対する除菌効果、について概説した。

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その他リンク： https://search.jamas.or.jp/default/link?pub_year=2023&ichushi_jid=J05115&link_issn=&doc_id=20230216410011&doc_link_id=%2Fdy6jahel%2F2023%2F002402%2F019%2F0154-0159%26dl%3D0&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fdy6jahel%2F2023%2F002402%2F019%2F0154-0159%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

記述言語：日本語   出版者・発行元：(一社)日本消化管学会  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Methods in molecular biology (Clifton, N.J.)  

Antimicrobial-resistant (AMR) bacterial infections caused by clinically important bacteria, including ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) and mycobacteria (Mycobacterium tuberculosis and nontuberculous mycobacteria), have become a global public health threat. Their epidemic and pandemic clones often accumulate useful accessory genes in their genomes, such as AMR genes (ARGs) and virulence factor genes (VFGs). This process is facilitated by horizontal gene transfer among microbial communities via mobile genetic elements (MGEs), such as plasmids and phages. Nanopore long-read sequencing allows easy and inexpensive analysis of complex bacterial genome structures, although some aspects of sequencing data calculation and genome analysis methods are not systematically understood. Here we describe the latest and most recommended experimental and bioinformatics methods available for the construction of complete bacterial genomes from nanopore sequencing data and the detection and classification of genotypes of bacterial chromosomes, ARGs, VFGs, plasmids, and other MGEs based on their genomic sequences for genomic epidemiological analysis of AMR bacteria.

DOI： 10.1007/978-1-0716-2996-3_16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Clinical Microbiology  

Despite frequent identification of plasmids carrying carbapenemase genes, the transfer of plasmids carrying carbapenemase genes is not well recognized in clinical settings because of technical limitations. To investigate the detailed mechanisms of the spread of carbapenem-resistant Enterobacteriaceae (CRE), we performed multifaceted genomic surveillance of CRE isolates in Thailand and analyzed their plasmidome. We analyzed 371 Enterobacteriaceae isolates carrying blaNDM-1 and 114 Enterobacteriaceae isolates carrying blaNDM-5 obtained from clinical samples of 473 patients in 11 representative hospitals located in six provinces in Thailand between 2012 and 2017. The complete structures of plasmids carrying blaNDM and chromosomal phylogeny were determined by combining Southern blotting hybridization analysis and our previously performed whole-genome short-read sequencing data. Dissemination of the blaNDM-5 gene among the Enterobacteriaceae isolates in Thailand was mainly owing to the nationwide clonal spread of Escherichia coli ST410 and regional clonal spreads of Escherichia coli ST361 and ST405. Analysis of blaNDM-1-carrying isolates revealed nationwide dissemination of two specific plasmids and nationwide clonal dissemination of Klebsiella pneumoniae ST16 accompanied with regional disseminations of three distinctive K. pneumoniae clones (ST231, ST14, and ST147) with different plasmids. Dissemination of CRE carrying blaNDM in Thailand is mainly based on nationwide clonal expansions of E. coli ST410 carrying blaNDM-5 and K. pneumoniae ST16 carrying blaNDM-1, nationwide dissemination of two distinctive plasmids carrying blaNDM-1, and accumulation of clonal expansions in regional areas. Although the overuse of antibiotics can promote CRE dissemination, the limited variety of transmitters highlights the importance of preventing horizontal dissemination among patients.

DOI： 10.1128/jcm.01080-22

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1101/2022.11.18.517170

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：iScience  

Helicobacter suis, hosted by hogs, is the most prevalent gastric non-Helicobacter pylori Helicobacter species found in humans. Recent studies have suggested that H. suis infection has caused many cases of gastric disease, but the transmission route from hogs remains unclear. Diagnostic methods based on H. suis urease activity often yield negative results, and there is no reliable method for diagnosing H. suis infection in clinical practice without gastric biopsy specimens. This study presents the world's first use of whole-bacterial cell ELISA to simultaneously assess H. suis and H. pylori infections. The ELISAs showed high accuracy, with an area under the ROC curve of 0.96, 100% sensitivity, 92.6% specificity, 76.9% positive predictive value, and 100% negative predictive value for the H. suis test, and an area under the ROC curve of 0.92, 88.2% sensitivity, 87.5% specificity, 65.2% positive predictive value, and 96.6% negative predictive value for the H. pylori test.

DOI： 10.1101/2022.10.10.22280809

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記述言語：英語   出版者・発行元：Cold Spring Harbor Laboratory  

Antimicrobial resistance genes (ARGs) are associated with mobile genetic elements (MGEs) that conscript useful genes into the human-microbe and microbe-microbe battlefields. Thus, under intense selective pressure, ARGs have been constantly adapting and evolving, spreading among microbes. tmexCD-toprJ gene clusters, which encode resistance-nodulation-cell division (RND)-type efflux pumps, confer multidrug-resistance to clinically important antimicrobials, including tigecycline. Noteworthily, these gene clusters have emerged in gram-negative bacteria in humans, animals, and the environment worldwide by MGE-mediated transfer. Here we show a hidden MGE, strand-biased circularizing integrative element (SE), that is recently recognized to mediate transpositions of ARGs, associated with the spread of tmexCD-toprJ gene clusters. We identified multidrug-resistant isolates of Aeromonas species in a water environment in Vietnam that harbored multiple copies of tmexCD-toprJ in their chromosomes that were associated with SEs. In particular, Aeromonas hydrophila NUITM-VA1 was found to harbor two copies of a novel variant of tmexC3.3D3.3-topJ1 within cognate SEs, whereas Aeromonas caviae NUITM-VA2 harbored four copies of a novel variant of tmexC2D2.3-topJ2 within cognate SEs. Based on the nature of SE to incorporate a neighboring sequence into the circular form and reinsert it into target sites during transposition, we identified the order of intragenomic movements of tmexCD-toprJ gene clusters. Altogether, our findings suggest that most known subgroups of tmexCD-toprJ and their subvariants underwent transpositions among bacterial chromosomes and plasmids via SEs. Hence, a tmexCD-toprJ gene cluster ancestor may have been initially mobilized via SE, subsequently spreading among bacteria and evolving in new hosts.

DOI： 10.1101/2022.09.22.508988

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記述言語：英語   出版者・発行元：Cold Spring Harbor Laboratory  

The emergence of the mobile resistance-nodulation-division (RND)-type efflux pump tmexCD-toprJ gene cluster that confers multidrug resistance (MDR), including tigecycline resistance, in gram-negative bacteria poses a global public health threat. However, the spread of such clinically important antimicrobial resistance genes (ARGs) in the natural environment has not yet been well investigated. In this study, we investigated MDR aquatic bacteria in Vietnam. A carbapenem- and tigecycline-resistant Shewanella xiamenensis isolate NUITM-VS2 was obtained from urban drainage in Hanoi, Vietnam, in October 2021. S. xiamenensis NUITM-VS2 showed resistance to most antimicrobials tested, including tigecycline, tetracyclines, carbapenems, cephalosporins, fluoroquinolone, and aminoglycosides. Whole-genome analysis was performed by long- and short-read sequencing, resulting in the complete genome sequence consisting of one chromosome and five plasmid sequences. ARGs and plasmid replicons in the genome were detected using ResFinder with the custom ARG database, including all known tigecycline resistance genes, and PlasmidFinder, respectively. A 152.2-kb IncC plasmid, pNUITM-VS2_2, co-carried two mobile tigecycline resistance genes, tet(X4) and tmexC3.1D3.1-toprJ1. In addition, a 24.8-kb untypeable plasmid, pNUITM-VS2_4, carried the carbapenemase gene blaNDM-1. pNUITM-VS2_2 was transferred to Escherichia coli by conjugation, which simultaneously conferred high-level resistance against many antimicrobials, including tigecycline. To the best of our knowledge, this is the first report of the detection of the mobile RND-type efflux pump gene cluster tmexCD-toprJ in Shewanella species. Our results provide genetic evidence of the complexity of the dynamics of clinically important ARGs among aquatic bacteria, which could be important reservoirs for ARGs in the natural environment.

DOI： 10.1101/2022.09.15.508154

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Oxford University Press (OUP)  

<jats:title>Abstract</jats:title>
<jats:sec>
<jats:title>Background</jats:title>
<jats:p>Klebsiella pneumoniae ST258 and ST11 carrying blaKPC are among the most widespread carbapenem-resistant K. pneumoniae strains worldwide. Our carbapenem-resistant Enterobacteriaceae surveillance in Thailand revealed a nationwide dissemination of K. pneumoniae ST16 isolates carrying blaNDM-1 and blaOXA-232.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Objectives</jats:title>
<jats:p>To analyse the genomic details of this nationwide dissemination by focusing on plasmids and virulence factors.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Methods</jats:title>
<jats:p>Using WGS data of 119 K. pneumoniae ST16 isolates carrying blaNDM-1 obtained in our previous surveillance study, clonality of chromosomes and plasmids of the isolates with carriage of virulence factors was evaluated.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Results</jats:title>
<jats:p>Of the 119 isolates, 111 carried plasmid pKP151_NDM1, and all 104 isolates harbouring blaOXA-232 carried plasmid pKP151_OXA232. These 104 K. pneumoniae ST16 isolates showing chromosomal clonality possessed both pKP151_NDM1 and pKP151_OXA232, demonstrating clonal dissemination of K. pneumoniae ST16 with these plasmids. The isolates had essentially similar virulence factors as those of K. pneumoniae ST16 clones carrying blaKPC, which were recently reported as highly invasive clones in Brazil.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Conclusions</jats:title>
<jats:p>The potential global dissemination of these invasive clones with resistance to several antibiotics highlights the importance of appropriate monitoring and strict standard precautions.</jats:p>
</jats:sec>

DOI： 10.1093/jacamr/dlac084

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

Helicobacter suisはブタが自然宿主と考えられている細菌で、ヒト胃での持続感染は胃粘膜関連リンパ組織(MALT)リンパ腫などの胃関連疾患の発症に関連していることが指摘されている。H.suisは過去にブタから単離培養されていたが、ヒトからの分離はなされていなかったため、ヒトにおいてH.suis感染と胃関連疾患発症の因果関係の証明は不完全なままとなっていた。本研究では、われわれはヒト胃から直接的にH.suisを分離し、in vitroで培養することに成功した。H.suisはHelicobacter pyloriと異なり、中性pHで著しく生存率が低下するため、胃生検の輸送には低pH培地を用いた。最終的に胃MALTリンパ腫を含む胃関連疾患患者3名からH.suisを分離することができた。患者全員において抗菌治療によりH.suisの除菌に成功し、内視鏡所見、病理組織学的所見が有意に改善された。H.suis臨床分離株をマウスに経口感染させると、胃粘膜で持続感染し、胃局所および全身性の炎症反応が惹起され、感染4ヵ月後には胃粘膜上皮化生が進行することが確認された。また、感染マウスの胃からH.suisが分離されたことから、本菌はヒトの胃関連疾患の起因菌としてコッホの原則を満たす形で初めて証明された。臨床分離株を用いたH.suisの比較ゲノム解析により、各分離株の染色体にはIV型分泌機構関連遺伝子などの株特異的な病原因子をコードする可塑性の高い領域が存在していること、ヒトとブタの分離株は遺伝的に非常に似ており、ブタ-ヒト感染の可能性があることが示唆された。(著者抄録)

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

Helicobacter suisはブタが自然宿主と考えられている細菌で、ヒト胃での持続感染は胃粘膜関連リンパ組織(MALT)リンパ腫などの胃関連疾患の発症に関連していることが指摘されている。H.suisは過去にブタから単離培養されていたが、ヒトからの分離はなされていなかったため、ヒトにおいてH.suis感染と胃関連疾患発症の因果関係の証明は不完全なままとなっていた。本研究では、われわれはヒト胃から直接的にH.suisを分離し、in vitroで培養することに成功した。H.suisはHelicobacter pyloriと異なり、中性pHで著しく生存率が低下するため、胃生検の輸送には低pH培地を用いた。最終的に胃MALTリンパ腫を含む胃関連疾患患者3名からH.suisを分離することができた。患者全員において抗菌治療によりH.suisの除菌に成功し、内視鏡所見、病理組織学的所見が有意に改善された。H.suis臨床分離株をマウスに経口感染させると、胃粘膜で持続感染し、胃局所および全身性の炎症反応が惹起され、感染4ヵ月後には胃粘膜上皮化生が進行することが確認された。また、感染マウスの胃からH.suisが分離されたことから、本菌はヒトの胃関連疾患の起因菌としてコッホの原則を満たす形で初めて証明された。臨床分離株を用いたH.suisの比較ゲノム解析により、各分離株の染色体にはIV型分泌機構関連遺伝子などの株特異的な病原因子をコードする可塑性の高い領域が存在していること、ヒトとブタの分離株は遺伝的に非常に似ており、ブタ-ヒト感染の可能性があることが示唆された。(著者抄録)

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley  

<jats:title>Abstract</jats:title><jats:sec><jats:title>Background</jats:title><jats:p>Eradication treatment for <jats:italic>Helicobacter pylori</jats:italic> gastritis is covered by national health insurance since 2013 in Japan. However, eradication failure due to the increase of antimicrobial resistance has become a serious problem. The present study aims to establish a reference panel of Japanese <jats:italic>H. pylori</jats:italic> strains for antimicrobial susceptibility testing.</jats:p></jats:sec><jats:sec><jats:title>Method</jats:title><jats:p>A total of 28 strains were collected from 4 medical facilities in Japan. Antimicrobial susceptibility tests (ASTs) to clarithromycin (CLR), amoxicillin (AMX), and metronidazole (MNZ), were used to select standard reference strains. Complete genome sequences were also determined.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Three <jats:italic>H. pylori</jats:italic> strains (JSHR3, JSHR6 and JSHR31) were selected as standard reference strains by the Japanese Society for <jats:italic>Helicobacter</jats:italic> Research (JSHR). The minimum inhibitory concentrations (MICs) of the antibiotics against these 3 strains by agar dilution method with Brucella‐based horse‐serum‐containing agar medium were as follows: JSHR3 (CLR 16 μg/ml, AMX 0.032 μg/ml and MNZ 4 μg/ml), JSHR6 (CLR 0.016 μg/ml, AMX 0.032 μg/ml and MNZ 4 μg/ml), and JSHR31 (CLR 16 μg/ml, AMX 1 μg/ml and MNZ 64 μg/ml).</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>A reference panel of <jats:italic>H. pylori</jats:italic> JSHR strains was established. The panel consisted of JSHR6, which was antibiotic‐susceptible, JSHR3, which was CLR‐resistant, and JSHR31, which was multi‐resistant. This reference panel will be essential for standardized ASTs before the optimal drugs are selected for eradication treatment.</jats:p></jats:sec>

DOI： 10.1111/hel.12874

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記述言語：英語   出版者・発行元：Oxford University Press (OUP)  

<jats:title>Abstract</jats:title>
<jats:p>We describe a case of bacteremia in a human immunodeficiency virus–infected patient caused by a Bordetella pertussis strain lacking 2 major virulence factors, filamentous hemagglutinin and fimbriae. Although B pertussis bacteremia is uncommon, physicians should be aware that even attenuated B pertussis strains can cause invasive infection in immunocompromised patients.</jats:p>
<jats:p>Bordetella pertussis is a gram-negative coccobacillus that causes a severe paroxysmal coughing disease known as whooping cough or pertussis. Bordetella pertussis colonizes the epithelial cells of the human respiratory tract, and the organisms are typically isolated from nasopharynx. We describe a case of B pertussis bacteremia in a patient with human immunodeficiency virus (HIV) infection. Interestingly, the isolate recovered from blood culture did not produce the major virulence factors, filamentous hemagglutinin (FHA) and fimbriae (FIM). Previously, 3 cases of B pertussis bacteremia were reported in the literature. We discuss the features of B pertussis bacteremia.</jats:p>

DOI： 10.1093/ofid/ofac020

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Global Antimicrobial Resistance  

Objectives: Tigecycline resistance mediated by the mobile tigecycline-inactivating enzyme gene tet(X) in Gram-negative bacteria is an emerging concern for global public health. However, limited information is available on the distribution of tet(X) in the natural environment. In this study, we investigated the presence of tet(X) in environmental Gram-negative bacteria. Methods: A carbapenem- and tigecycline-resistant Shewanella xiamenensis isolate (NUITM-VS1) was obtained from an urban drainage in Hanoi, Vietnam, in March 2021. Whole-genome sequencing analysis was performed by long- and short-read sequencing, resulting in a complete genome sequence. Antimicrobial resistance genes (ARGs) in the genome were detected based on the custom ARG database, including all known tigecycline resistance genes. Results: Shewanella xiamenensis isolate NUITM-VS1 harboured the tet(X4) gene and the blaOXA-48 carbapenemase gene on the chromosome. tet(X4) was flanked by IS91 family transposase genes, suggesting that the acquisition of tet(X4) was mediated by this mobile gene element (MGE), whereas no MGE was found surrounding blaOXA-48, consistent with previous findings that blaOXA-48-like β-lactamase genes are species-specific intrinsic ARGs in Shewanella spp. Conclusion: To the best of our knowledge, this is the first report of a tet(X4)-harbouring Shewanella sp. isolate. Our results provide genetic evidence of the complexity of the dynamics of clinically important ARGs among bacteria in the water environment.

DOI： 10.1016/j.jgar.2021.12.022

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記述言語：日本語   出版者・発行元：(一社)日本胃癌学会  

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記述言語：日本語   出版者・発行元：(一社)日本胃癌学会  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Oxford University Press (OUP)  

<jats:title>Abstract</jats:title>
<jats:sec>
<jats:title>Objectives</jats:title>
<jats:p>To characterize Acinetobacter baumannii OCU_Ac16a, a clinical isolate co-harbouring three acquired carbapenemase genes, blaNDM-1, blaTMB-1, and blaOXA-58, and assess the clinical significance of so-called multiple-carbapenemase producers.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Methods</jats:title>
<jats:p>OCU_Ac16a and its close relative, OCU_Ac16b, which lacks the blaNDM-1, were isolated from sputum cultures of a patient at Osaka City University Hospital. We subjected these strains to whole-genome analysis, particularly focusing on the genetic context of each carbapenemase gene. The transmissibility and functionality of each carbapenemase gene were analysed by conjugation and transformation experiments and antimicrobial susceptibility tests.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Results</jats:title>
<jats:p>bla TMB-1 was located in a class 1 integron on the chromosome, whereas blaNDM-1 and blaOXA-58 were found on plasmids named pOCU_Ac16a_2 and pOCU_Ac16a_3, respectively. pOCU_Ac16a_2 (which exhibited highly efficient self-transmissibility) and pOCU_Ac16a_3 (which did not show transmissibility but could be introduced into another A. baumannii strain via electroporation) could both confer carbapenem resistance (MICs ≥512 and ≥32 mg/L, respectively) on the recipient strain. The functionality of blaTMB-1 was evident from the high resistance of OCU_Ac16b to ceftazidime and cefepime (MICs ≥256 and 48 mg/L, respectively), and the high resistance of OCU_Ac16a to cefiderocol (MIC 32 mg/L) could be explained by the additive effect of blaNDM-1 and blaTMB-1.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Conclusions</jats:title>
<jats:p>Our data revealed the genomic organization of OCU_Ac16a and demonstrated that all the carbapenemase genes are functional, each contributing to the extremely high broad-spectrum resistance of OCU_Ac16a to β-lactams. As multiple-carbapenemase producers can be serious health threats as drug-resistant pathogens and disseminators of carbapenemase genes, close attention should be paid to their emergence.</jats:p>
</jats:sec>

DOI： 10.1093/jacamr/dlab191

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Global Antimicrobial Resistance  

Objectives: Mobile colistin resistance (mcr) genes are widely distributed around the world. To date, ten major variants of mcr genes are known (mcr-1 to mcr-10). However, only a few instances of Enterobacterales isolates harbouring mcr genes other than mcr-1 have been reported in Vietnam. The aim of this study was to investigate mcr-harbouring antimicrobial-resistant Enterobacterales isolates in Vietnam. Methods: Two mcr-9-harbouring Enterobacter hormaechei clinical isolates (NIHE14-1904 and MH17-539M) were obtained from medical institutions in Hanoi, Vietnam, in 2014 and 2017, respectively. Their genomes and plasmid sequences were analysed by short-read and long-read sequencing. Subsequently, comparative sequence analysis of their mcr-9-carrying plasmids was performed. Results: Strains NIHE14-1904 and MH17-539M belonged to sequence types ST916 and ST66, respectively, according to the Enterobacter cloacae multilocus sequence typing (MLST) scheme. NIHE14-1904 and MH17-539M harboured the mcr-9 gene on similar IncHI2 plasmids, namely pNIHE14-1904-mcr9 (373.1 kb) and pMH17-539M-mcr9 (289.3 kb), respectively. These plasmids were also highly identical to widespread IncHI2 plasmids that are often associated with mcr genes. Conclusion: For the first time, mcr-9-harbouring Enterobacterales isolates were detected in Vietnam, which carried mcr-9 on IncHI2 plasmids. The prevalence of such plasmids needs to be monitored in the future owing to their high dissemination.

DOI： 10.1016/j.jgar.2021.09.012

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Global Antimicrobial Resistance  

Objectives:: The incidence of carbapenem resistance among nosocomial Gram-negative bacteria in Vietnam is high and increasing, including among Enterobacterales. In this study, we assessed the presence of one of the main carbapenemase genes, blaKPC, among carbapenem-resistant Enterobacterales (CRE) from four large hospitals in Hanoi, Vietnam, between 2010 and 2015, and described their key molecular characteristics. Methods:: KPC-producing Enterobacterales were detected using conventional PCR and were further analysed using S1 nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting and whole-genome sequencing (WGS) for sequence typing and genetic characterisation. Results:: blaKPC genes were detected in 122 (20.4%) of 599 CRE isolates. blaKPC-carrying plasmids were diverse in size. Klebsiella pneumoniae harbouring blaKPC genes belonged to ST15 and ST11, whereas KPC-producing Escherichia coli showed more diverse sequence types including ST3580, ST448, ST709 and ST405. Genotypic relationships supported the hypothesis of circulation of a population of ‘resident’ resistant bacteria in one hospital through the years and of transmission among these hospitals via patient transfer. WGS results revealed co-carriage of several other antimicrobial resistance genes and three different genetic contexts of blaKPC-2. Among these, the combination of ISEcp1–blaCTX-M and ISKpn27–blaKPC–ΔISKpn6 on the same plasmid is reported for the first time. Conclusion:: We describe the dissemination of blaKPC-expressing Enterobacterales in four large hospitals in Hanoi, Vietnam, since 2010, which may have started earlier, along with their resistance patterns, sequence types, genotypic relationship, plasmid sizes and genetic context, thereby contributing to the overall picture of the antimicrobial resistance situation in Enterobacterales in Vietnam.

DOI： 10.1016/j.jgar.2021.09.007

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Hospital Infection  

Background: The impact of the coronavirus disease (COVID-19) pandemic on antimicrobial resistance (AMR) is a major concern. Aim: To compare the number of patients and isolation rate of antimicrobial-resistant bacteria before and after the beginning of the COVID-19 pandemic using the comprehensive national surveillance data. Methods: We utilized comprehensive surveillance data, collected in the Japan Nosocomial Infections Surveillance programme, which included a total of 16.7 million samples of 5.9 million tested patients from >1300 hospitals. We compared the number of patients and isolation rate of five bacteria between 2019 and 2020, including antimicrobial-susceptible and -resistant bacteria of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. Findings: The number of patients and isolation rate of S. aureus and meticillin-resistant S. aureus decreased slightly; those of S. pneumoniae and penicillin-resistant S. pneumoniae decreased by 60%; and those of third-generation cephalosporin-resistant K. pneumoniae increased. The isolation rate of the remaining bacteria apparently increased, although the number of patients decreased. This was due to a substantial decrease in the total number of tested patients (the denominator of the isolation rate), which was larger than that of the number of patients (the numerator of the isolation rate). Consistent results were obtained when the same data were re-aggregated using the procedure of the World Health Organization Global Antimicrobial Resistance Surveillance System, demonstrating the general importance of this problem. Conclusion: Surveillance data during the COVID-19 pandemic must be carefully interpreted based on examination of the numerator, denominator and background factors that affect the denominator.

DOI： 10.1016/j.jhin.2021.09.011

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Oxford University Press (OUP)  

<jats:title>Abstract</jats:title>
<jats:sec>
<jats:title>Background</jats:title>
<jats:p>MDR bacteria including carbapenem-resistant Pseudomonas aeruginosa are recognized as an important cause of hospital-acquired infections worldwide. This investigation seeks to determine the molecular characterization and antibiotic resistance genes associated with carbapenem-resistant P. aeruginosa.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Methods</jats:title>
<jats:p>We conducted WGS and phylogenetic analysis of 72 carbapenem-resistant P. aeruginosa isolated from hospital-acquired infection patients from August 2011 to March 2015 in three major hospitals in Hanoi, Vietnam.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Results</jats:title>
<jats:p>We identified three variants of IMP gene, among which blaIMP-15 was the most frequent (n = 34) in comparison to blaIMP-26 (n = 2) and blaIMP-51 (n = 12). We observed two isolates with imipenem MIC &gt;128 mg/L that co-harboured blaIMP-15 and blaDIM-1 genes and seven isolates (imipenem MIC &gt; 128 mg/L) with a blaKPC-1 gene from the same hospital. MLST data shows that these 72 isolates belong to 18 STs and phylogenetic tree analysis has divided these isolates into nine groups.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Conclusions</jats:title>
<jats:p>Our results provide evidence that not only blaIMP-26 but other IMP variants such as blaIMP-15 and blaIMP-51 genes and several STs (ST235, ST244, ST277, ST310, ST773 and ST3151) have been disseminating in healthcare settings in Vietnam. In addition, we report the emergence of two isolates belonging to ST1240 and ST3340 that harboured two important carbapenemase genes (blaIMP-15 and blaDIM-1) and seven isolates belonging to ST3151 of P. aeruginosa that carried the blaKPC-1 gene in Vietnam, which could potentially cause serious restricted availability of treatment options in healthcare settings.</jats:p>
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DOI： 10.1093/jacamr/dlab103

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

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記述言語：日本語   出版者・発行元：(一社)日本ヘリコバクター学会  

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記述言語：日本語   出版者・発行元：緑膿菌感染症研究会  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:p>
Tigecycline is important as a last-resort antimicrobial and effective against antimicrobial-resistant bacteria, such as carbapenem-producing
<jats:italic>Enterobacterales</jats:italic>
(CPE), whose infections are difficult to treat with antimicrobials. Since 2019, mobile tigecycline resistance genes,
<jats:italic>tet</jats:italic>
(X) and
<jats:italic>tmexCD-toprJ</jats:italic>
, and their variants have been reported mainly from China, and it has become important to understand their epidemiological situation and detailed genetic mechanisms.
</jats:p>

DOI： 10.1128/mSphere.00592-21

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Infectious Diseases  

Background: Antimicrobial resistance (AMR) is a global health problem. However, quantitative evaluation of its disease burden is challenging. This study aimed to estimate the disease burden of bloodstream infections (BSIs) caused by major antimicrobial-resistant bacteria in Japan between 2015 and 2018 in terms of disability-adjusted life-years (DALYs). Methods: DALYs of BSIs caused by nine major antimicrobial-resistant bacteria in Japan were estimated using comprehensive national surveillance data of all routine bacteriological test results from more than 1400 hospitals between 2015 and 2018. The methodology of Cassini et al. was modified to enable comparison of the present results with those in other countries. Results: It was estimated that 137.9 [95% uncertainty interval (UI) 130.7⿿145.2] DALYs per 100,000 population were attributable to BSIs caused by nine antimicrobial-resistant bacteria in 2018. Methicillin-resistant Staphylococcus aureus (MRSA), fluoroquinolone-resistant Escherichia coli (FQREC) and third-generation cephalosporin-resistant E. coli (3GREC) accounted for 87.2% overall. The burden did not decrease during the study period and was highest in people aged ⿥65 years. Conclusion: The results revealed, for the first time, the disease burden of BSIs caused by nine major antimicrobial-resistant bacteria in Japan. The estimated disease burden associated with AMR in Japan is substantial and has not begun to decrease. Notably, the burden from FQREC and 3GREC has increased steadily, and that from MRSA is larger in Japan than in the European Union/European Economic Area, whereas the burden from other bacteria is comparatively small. These results are expected to provide useful information for healthcare policy makers for prioritizing interventions for AMR./

DOI： 10.1016/j.ijid.2021.05.018

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS Pathogens  

Length control is a fundamental requirement for molecular architecture. Even small wall-less bacteria have specially developed macro-molecular structures to support their survival. Mycoplasma pneumoniae, a human pathogen, forms a polar extension called an attachment organelle, which mediates cell division, cytadherence, and cell movement at host cell surface. This characteristic ultrastructure has a constant size of 250–300 nm, but its design principle remains unclear. In this study, we constructed several mutants by genetic manipulation to increase or decrease coiled-coil regions of HMW2, a major component protein of 200 kDa aligned in parallel along the cell axis. HMW2-engineered mutants produced both long and short attachment organelles, which we quantified by transmission electron microscopy and fluorescent microscopy with nano-meter precision. This simple design of HMW2 acting as a molecular ruler for the attachment organelle should provide an insight into bacterial cellular organization and its function for their parasitic lifestyles.

DOI： 10.1371/journal.ppat.1009621

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microorganisms  

Many pathogenic bacteria, including Escherichia coli and Vibrio cholerae, can become viable but nonculturable (VBNC) following exposure to specific stress conditions. Corynebacterium diphtheriae, a known human pathogen causing diphtheria, has not previously been shown to enter the VBNC state. Here, we report that C. diphtheriae can become VBNC when exposed to low temperatures. Morphological differences in culturable and VBNC C. diphtheriae were examined using scanning electron microscopy. Culturable cells presented with a typical rod-shape, whereas VBNC cells showed a distorted shape with an expanded center. Cells could be transitioned from VBNC to culturable following treatment with catalase. This was further evaluated via RNA sequence-based transcriptomic analysis and reverse-transcription quantitative PCR of culturable, VBNC, and resuscitated VBNC cells following catalase treatment. As expected, many genes showed different behavior by resuscitation. The expression of both the diphtheria toxin and the repressor of diphtheria toxin genes remained largely unchanged under all four conditions (culturable, VBNC, VBNC after the addition of catalase, and resuscitated cells). This is the first study to demonstrate that C. diphtheriae can enter a VBNC state and that it can be rescued from this state via the addition of catalase. This study helps to expand our general understanding of VBNC, the pathogenicity of VBNC C. diphtheriae, and its environmental survival strategy.

DOI： 10.3390/microorganisms9050927

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BMC Infectious Diseases  

Background: The association between the frequency of surgeries and the incidence of surgical site infections (SSIs) has been reported for various surgeries. However, no previous study has explored this association among video-assisted thoracic surgeries (VATS). Hence, we aimed to investigate the association between the frequency of surgeries and SSI in video-assisted thoracic surgeries. Methods: We analyzed the data of 26,878 thoracic surgeries, including 21,154 VATS, which were collected during a national surveillance in Japan between 2014 and 2018. The frequency of surgeries per hospital department was categorized into low (< 50/year), moderate (50–100/ year), and high (> 100/year). Chi-squared test or Fisher’s exact test was used for discrete explanatory variables, whereas Wilcoxon’s rank-sum test or Kruskal-Wallis test was used for continuous explanatory variables. Univariate analysis of the department groups was conducted to explore confounding factors associated with both SSIs and the department groups. We used a multiple logistic regression model focusing on VATS and stratified by the National Nosocomial Infections Surveillance System (NNIS) risk index. Results: The rates of SSIs in the hospital groups with low, moderate, and high frequency of surgeries were 1.39, 1.05, and 1.28%, respectively. In the NNIS risk index 1 stratum, the incidence of SSIs was significantly lower in the moderate-frequency of surgeries group than that in the other groups (odds ratio [OR]: vs. low-frequency of surgeries: 2.48 [95% confidence interval [CI]: 1.20–5.13], P = 0.0143; vs. high-frequency of surgeries: 2.43 [95% CI: 1.44–4.11], P = 0.0009). In the stratum of NNIS risk indices 2 and 3, the incidence of SSI was significantly higher in the low-frequency of surgeries group (OR: 4.83, 95% CI: 1.47–15.93; P = 0.0095). Conclusion: The result suggests that for departments with low-frequency of surgeries, an increase in the frequency of surgeries to > 50 per department annually potentially leads to a decrease in the incidence of SSIs. This occurs through an increase in the experience of the departmental surgeons and contributes to the improvement of VATS outcomes in thoracic surgeries.

DOI： 10.1186/s12879-021-06050-6

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

Genes conferring carbapenem resistance have spread worldwide among gram-negative bacteria. Subtyping of these genes has epidemiological value due to the global cross-border movement of people. Subtyping of blaIMP genes that frequently detected in Japan appears to be important in public health settings; however, there are few useful tools for this purpose. We developed a subtyping screening tool based on PCR direct sequencing, which targets the internal sequences of almost all blaIMP genes. The tool used bipartite multiplex primers with M13 universal sequences at the 5'-end. According to in silico analysis, among the 78 known IMP-type genes, except for blaIMP-81, 77 detected genes were estimated to be differentiated. In vitro evaluation indicated that sequences of amplicons of IMP-1, IMP-6, IMP-7, and IMP-20 templates were identical to their respective subtypes. Even if the amplicons were small or undetectable through the first PCR, sufficient amplicons for DNA sequencing were obtained through a second PCR using the M13 universal primers. In conclusion, our tool can be possibly used for subtype screening of blaIMP, which is useful for the surveillance of bacteria with blaIMP in clinical and public health settings or environmental fields.

DOI： 10.7883/yoken.JJID.2020.926

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Proceedings of the National Academy of Sciences  

<jats:title>Significance</jats:title>
<jats:p>
This study directly demonstrates virulence-associated features of
<jats:italic>Helicobacter suis</jats:italic>
infection using isolates obtained from human stomachs.
<jats:italic>H. suis</jats:italic>
is the second-most-prevalent
<jats:italic>Helicobacter</jats:italic>
species in the human stomach, but diagnostic tests for
<jats:italic>Helicobacter pylori</jats:italic>
often fail to diagnose
<jats:italic>H. suis. H. suis</jats:italic>
does not have orthologs of either cytotoxin-associated gene A (CagA) or vacuolating cytotoxin A (VacA), two major virulence factors in
<jats:italic>H. pylori</jats:italic>
, suggesting that the molecular pathogenicity of
<jats:italic>H. suis</jats:italic>
and clinical manifestations caused by
<jats:italic>H. suis</jats:italic>
infection are different from those caused by
<jats:italic>H. pylori</jats:italic>
. This study paves the way for epidemiological research aimed at identifying the causal relationships between
<jats:italic>H. suis</jats:italic>
and gastric diseases and developing diagnostic methods.
</jats:p>

DOI： 10.1073/pnas.2026337118

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

Japan Nosocomial Infections Surveillance (JANIS) is one of the largest national antimicrobial resistance (AMR) surveillance systems in the world. The JANIS Clinical Laboratory division collects comprehensive specimen-based data from diagnostic microbiology laboratories of participating hospitals to monitor the isolation rate of 11 major bacteria and specific AMR bacteria, and creates antibiograms of approximately 20 bacterial species. Data on the JANIS web database system are also annually tabulated and shared with the WHO Global Antimicrobial Resistance Surveillance System. To create a network of international AMR surveillance systems among Asian countries, Japan is developing an international web database system named ASIan Antimicrobial Resistance Surveillance Network (ASIARS-Net) based on the JANIS system; ASIARS-Net is an open-source database and confidentially available at almost no cost. JANIS continues to evolve in multiple directions; some are discussed at the end of this review.

DOI： 10.7883/yoken.JJID.2020.499

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

Multilocus variable-number tandem repeat analysis (MLVA) is widely used for genotyping of Bordetella pertussis, the causative bacteria for pertussis. However, MLVA genotyping is losing its discriminate power because prevalence of the epidemic MT27 strain (MLVA-27) is increasing worldwide. To address this, we developed a single nucleotide polymorphism (SNP) genotyping method for MT27 based on multiplexed single-base extension (SBE) assay. A total of 237 MT27 isolates collected in Japan during 1999–2018 were genotyped and classified into ten SNP genotypes (SG1 to SG10) with a Simpson’s diversity index (DI) of 0.79 (95% CI 0.76–0.82). Temporal trends showed a marked increase in the genotypic diversity in the 2010s: Simpson’s DI was zero in 1999–2004, 0.16 in 2005–2009, 0.83 in 2010–2014, and 0.76 in 2015–2018. This indicates that the SNP genotyping is applicable to the recently circulating MT27 strain. Additionally, almost all outbreak-associated MT27 isolates were classified into the same SNP genotypes for each outbreak. Multiplexed SBE assay allows for rapid and simple genotyping, indicating that the SNP genotyping can potentially be a useful tool for subtyping the B. pertussis MT27 strain in routine surveillance and outbreak investigations.

DOI： 10.1038/s41598-021-84409-0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

<jats:p>Tigecycline is a last-resort antimicrobial used to treat multidrug-resistant Gram-negative bacterial infections. One of the common antimicrobial resistance mechanisms is the efflux pump system composed of membrane protein complexes to excrete xenobiotic substrates. Recently, a novel gene cluster, <jats:italic>tmexCD1-toprJ1</jats:italic>, encoding the resistance–nodulation–cell division (RND) efflux pump was identified on plasmids in <jats:italic>
<jats:named-content content-type="species">
<jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.10789" xlink:type="simple">Klebsiella pneumoniae</jats:ext-link>
</jats:named-content>
</jats:italic> isolates in China. TMexCD1-TOprJ1 was found to be capable of excreting multiple antimicrobials, including tigecycline, which contributed to the strain's resistance. In this study, we identified <jats:italic>
<jats:named-content content-type="species">
<jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.10789" xlink:type="simple">K. pneumoniae</jats:ext-link>
</jats:named-content>
</jats:italic> isolates harbouring the <jats:italic>tmexCD1-toprJ1</jats:italic> genes outside of China for the first time. Two tigecycline-resistant <jats:italic>
<jats:named-content content-type="species">
<jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.10789" xlink:type="simple">K. pneumoniae</jats:ext-link>
</jats:named-content>
</jats:italic> isolates belonging to ST273 by multilocus sequence typing were collected from different patients in a medical institution in Hanoi, Vietnam, in 2015. Whole-genome sequence analysis revealed that these isolates harboured a 288.0 kb <jats:italic>tmexCD1-toprJ1</jats:italic>–carrying plasmid with IncFIB and IncHI1B replicons. The <jats:italic>tmexCD1-toprJ1</jats:italic> gene cluster was surrounded by several mobile gene elements, including IS<jats:italic>26</jats:italic>, and the plasmids had high sequence identity with that of <jats:italic>
<jats:named-content content-type="species">
<jats:ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.10789" xlink:type="simple">K. pneumoniae</jats:ext-link>
</jats:named-content>
</jats:italic> isolated in China. Our finding suggests that the horizontal spread of tigecycline resistance mediated by <jats:italic>tmexCD1-toprJ1</jats:italic>–carrying plasmids has occurred in Vietnam and other countries, and raises concern about the further global dissemination.</jats:p>

DOI： 10.1099/jmm.0.001320

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：BMC Infectious Diseases  

Background: The goals of the National Action Plan on Antimicrobial Resistance (AMR) of Japan include “implementing appropriate infection prevention and control” and “appropriate use of antimicrobials,” which are relevant to healthcare facilities. Specifically, linking efforts between existing infection control teams and antimicrobial stewardship programs was suggested to be important. Previous studies reported that human resources, such as full-time equivalents of infection control practitioners, were related to improvements in antimicrobial stewardship. Methods: We posted questionnaires to all teaching hospitals (n = 1017) regarding hospital countermeasures against AMR and infections. To evaluate changes over time, surveys were conducted twice (1st survey: Nov 2016, 2nd survey: Feb 2018). A latent transition analysis (LTA) was performed to identify latent statuses, which refer to underlying subgroups of hospitals, and effects of the number of members in infection control teams per bed on being in the better statuses. Results: The number of valid responses was 678 (response rate, 66.7%) for the 1st survey and 559 (55.0%) for the 2nd survey. More than 99% of participating hospitals had infection control teams, with differences in activity among hospitals. Roughly 70% had their own intervention criteria for antibiotics therapies, whereas only about 60 and 50% had criteria established for the use of anti-methicillin-resistant Staphylococcus aureus antibiotics and broad-spectrum antibiotics, respectively. Only 50 and 40% of hospitals conducted surveillance of catheter-associated urinary tract infections and ventilator-associated pneumonia, respectively. Less than 50% of hospitals used maximal barrier precautions for central line catheter insertion. The LTA identified five latent statuses. The membership probability of the most favorable status in the 2nd study period was slightly increased from the 1st study period (23.6 to 25.3%). However, the increase in the least favorable status was higher (26.3 to 31.8%). Results of the LTA did not support a relationship between increasing the number of infection control practitioners per bed, which is reportedly related to improvements in antimicrobial stewardship, and being in more favorable latent statuses. Conclusions: Our results suggest the need for more comprehensive antimicrobial stewardship programs and increased surveillance activities for healthcare-associated infections to improve antimicrobial stewardship and infection control in hospitals.

DOI： 10.1186/s12879-021-05921-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

Haemophilus influenzae is a small, nonmotile, non-spore-forming bacterium classified into 6 serotypes (a to f) and non-typeable strains that lack a capsule. Although H. influenzae serotype a (Hia) is prevalent in Canada, the United States, Brazil, Australia, across the African continent, and several other locations, it has not been reported in Japan thus far. Our case was of a 72-year-old Japanese man who sought medical consultation after presenting with chills, fever, and polyarthritis. Cultures of blood and synovial fluid from the left knee revealed H. influenzae infection. Diagnostic imaging showed poor contrast regions in both kidneys, fluid retention around both knee joints, the left shoulder joint, and both elbow joints. Subsequently, the patient was diagnosed with invasive H. influenzae infection accompanied by polyarthritis and renal infarction. 16S ribosomal RNA gene sequencing revealed that the bacterial strain was Hia. The patient was treated with antimicrobial agents and arthroscopic curettage. We present a case of invasive Hia infection accompanied by polyarthritis and renal infarction. To the best of our knowledge, this is the first case of Hia infection in Japan. The case is very rare considering that the disease occurred in an elderly patient who developed polyarthritis.

DOI： 10.1016/j.jiac.2021.01.017

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

Several outbreaks of trench fever caused by Bartonella quintana occurred in soldiers during World Wars I and II. Although trench fever cases have been decreasing worldwide, the disease was reported among the homeless population in developing and developed countries. The current prevalence of B. quintana infection in Japan is unclear. Blood and body louse (Pediculus humanus humanus) samples were obtained from homeless inpatients with body lice during emergency hospitalization in Tokyo from January 2013 to March 2015. Patients were tested for B. quintana infections using the culture method, polymerase chain reaction, and indirect immunofluorescence assay (IFA). Among the 29 patients tested, the presence of Bartonella spp. was confirmed by genomic sequencing of DNA extracted from two samples from blood culture performed for 15 out of 29 patients and from body louse samples of 20 patients (69%). Immunoglobulin G against B. quintana was detected in 10 patients (34.5%) at a cut-off titer of 1:256 in IFA. B. quintana infection was detected in samples obtained between 2013 and 2015 in Tokyo and needs to be on the list of differential diagnoses performed for febrile homeless individuals.

DOI： 10.7883/yoken.JJID.2020.505

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記述言語：英語   出版者・発行元：Research Square  

<title>Abstract</title>
<bold>Background:</bold> Multidrug-resistant bacteria including carbapenem resistant <italic>Pseudomonas aeruginosa </italic>are recognised as an important cause of hospital-acquired infections worldwide. To determine the molecular characterisation and antibiotic resistant genes associated with carbapenem-resistant <italic>P. aeruginosa</italic>. <bold>Methods:</bold> we conducted whole-genome sequencing and phylogenetic analysis of 72 carbapenem-resistant <italic>P. aeruginosa </italic>isolated from hospital-acquired infection patients from 2010 to 2015 in three major hospitals in Hanoi, Vietnam. <bold>Results:</bold> We identified three variants of IMP genes, among which IMP-15 gene was the most frequent (n= 34) in comparison to IMP-26 (n= 2) and IMP-51 (n=12). We observed two isolates with imipenem MIC &gt;128mg/L that co-harboured IMP-15 and DIM-1 genes and seven isolates (imipenem MIC&gt; 128mg/L) with KPC-1 gene from the same hospital. MLST data showed that sequence types (ST) of 72 isolates were classified into 18 STs and phylogenetic tree analysis divided these isolates into nine groups. <bold>Conclusion:</bold> Our results provide evidence that not only IMP-26, but other variants of IMPs like IMP-15 and IMP-51 genes and several STs (ST235, ST244, ST277, ST310, ST773 and ST3151) have been disseminated in health care settings in Vietnam. Also, we report the first finding in Vietnam that two isolates belonging to ST1240 and ST3340 harboured two important carbapenemase genes (IMP-15 and, DIM-1) and seven isolates belonging to ST3151 of <italic>P. aeruginosa </italic>carried the KPC-1 gene, which could be a potential cause of seriously restricted available treatment options in healthcare settings.

DOI： 10.21203/rs.3.rs-143498/v1

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その他リンク： https://www.researchsquare.com/article/rs-143498/v1.html

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Microbiology  

The rapid emergence of carbapenemase-producing gram-negative bacteria (CPGNB) is a global threat due to the high mortality of infection and limited treatment options. Although there have been many reports of CPGNB isolated from Southeast Asian countries, to date there has been no genetic analysis of CPGNB isolated from Cambodia. Sequence-based molecular epidemiological analysis enables a better understanding of the genotypic characteristics and epidemiological significance of antimicrobial-resistant (AMR) bacteria in each country, and allows countries to enact measures related to AMR issues. In this study, we performed on-site genomic epidemiological analysis of CPGNB isolated in Cambodia using a portable laboratory equipment called Bento Lab, which combines a PCR thermal cycler, microcentrifuge, gel electrophoresis apparatus, and LED transilluminator, along with the MinION nanopore sequencer. PCR targeting of major carbapenemase genes using Bento Lab revealed that two Escherichia coli isolates and one Acinetobacter baumannii isolate harbored carbapenemase genes: blaNDM, blaOXA–48, and blaOXA–23, respectively. The results of phenotypic diagnostic tests for CPGNB, such as the carbapenem inactivation method and double-disk diffusion test using a specific inhibitor of metallo-β-lactamases, were consistent with their AMR genotypes. Whole-genome sequencing analysis using MinION revealed that blaNDM–5 gene was carried on a 93.9-kb plasmid with IncFIA/IncFIB/IncFII/IncQ1 replicons, and blaOXA–181 gene was carried on a 51.5-kb plasmid with the IncX3 replicon in E. coli isolates. blaOXA–23 was encoded in two locations on the chromosome of A. baumannii. Plasmids carrying blaNDM–5 or blaOXA–181 in E. coli were highly structurally identical to plasmids prevalent in Enterobacterales in China and other countries, suggesting that they disseminated from a common evolutionary origin. Our findings demonstrate the potential impact of portable laboratory equipment on AMR bacteria research in hospitals and research centers with limited research facilities, and provide the first glimpse into the genomic epidemiology of CPGNB in Cambodia.

DOI： 10.3389/fmicb.2021.675463

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Resource Announcements  

Haemophilus influenzae causes severe infections such as pneumonia and meningitis. Here, we report the complete genome of H. influenzae type a strain TAMBA230, which was isolated in 2019 from a patient exhibiting bacteremia. This represents the first case in Japan of an H. influenzae type a strain associated with invasive infection.

DOI： 10.1128/MRA.01069-20

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Nature Communications  

Mycoplasma pneumoniae is a bacterial human pathogen that causes primary atypical pneumonia. M. pneumoniae motility and infectivity are mediated by the immunodominant proteins P1 and P40/P90, which form a transmembrane adhesion complex. Here we report the structure of P1, determined by X-ray crystallography and cryo-electron microscopy, and the X-ray structure of P40/P90. Contrary to what had been suggested, the binding site for sialic acid was found in P40/P90 and not in P1. Genetic and clinical variability concentrates on the N-terminal domain surfaces of P1 and P40/P90. Polyclonal antibodies generated against the mostly conserved C-terminal domain of P1 inhibited adhesion of M. pneumoniae, and serology assays with sera from infected patients were positive when tested against this C-terminal domain. P40/P90 also showed strong reactivity against human infected sera. The architectural elements determined for P1 and P40/P90 open new possibilities in vaccine development against M. pneumoniae infections.

DOI： 10.1038/s41467-020-18777-y

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Emerging Infectious Diseases  

Macrolide-resistant Bordetella pertussis emerged in Vietnam during 2016-2017. Direct analyses of swab samples from 10 patients with pertussis revealed a macrolide-resistant mutation, A2047G, in the 23S rRNA. We identified the MT104 genotype of macrolide-resistant B. pertussis (which is prevalent in mainland China) and its variants in these patients.

DOI： 10.3201/eid2610.201035

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Emerging Infectious Diseases  

We conducted molecular typing of a Corynebacterium ulcerans isolate from a woman who died in Japan in 2016. Genomic DNA modification might have affected the isolate's ribotyping profile. Multilocus sequence typing results (sequence type 337) were more accurate. Whole-genome sequencing had greater ability to discriminate lineages at high resolution.

DOI： 10.3201/eid2610.200086

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

There has been scarce evidence about deaths due to blood stream infection (BSI) in Japan so far. The main objective of this study is to understand the epidemiological trend of deaths caused by BSIs due to Staphylococcus aureus and Escherichia coli including Methicillin-resistant S. aureus (MRSA) and fluoroquinolone-resistant E. coli (FQREC) at national level. We annually estimated the number of BSI caused by S. aureus and E. coli between 2011 and 2017 across Japan using comprehensive data of bacterial culturing and drug susceptibilities collected in Japan Nosocomial Infection Surveillance (JANIS). The number of death was estimated by using BSI mortality obtained from previous studies in Japan. The number of BSI death attributable to S. aureus was estimated to 17,412 in 2011 and 17,157 in 2017, respectively, out of the whole population (126.8 million) in Japan. Among them, cases attributed to MRSA accounted for 5924 (34.0%) in 2011, and decreased to 4224 (24.6%) cases in 2017. On the other hand, the number of BSI death attributable to E. coli was estimated to 9044 in 2011 and increased to 14,016 in 2017. Among them, cases attributed to FQREC accounted for 2045 (22.6%) in 2011 and increased to 3915 (27.9%) cases in 2017. The number of BSI death attributable to MRSA has been decreasing and that attributable to FQREC has been increasing. This study provides the first annual estimate of disease burden of BSI caused by antimicrobial resistant (AMR) bacteria in Japan, and basis for formulating health policy to deal with AMR.

DOI： 10.1016/j.jiac.2019.10.017

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: bla bla bla bla bla and bla Of 70 blaIMP variants, 67 subtypes were simulated toKPC,be PCR-positiveIMP,NDM,basedVIM,on in silicoOXA-48-like,simulation andGES.the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.

DOI： 10.7883/yoken.JJID.2019.041

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

A multispecies outbreak of IMP-6 carbapenemase-producing Enterobacterales (IMP-6-CPE) occurred at an acute care hospital in Japan. This study was conducted to understand the mechanisms of IMP-6-CPE transmission by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing and whole-genome sequencing (WGS), and identify risk factors for IMP-6-CPE acquisition in patients who underwent abdominal surgery. Between July 2013 and March 2014, 22 hospitalized patients infected or colonized with IMP-6-CPE (Escherichia coli [n = 8], Klebsiella oxytoca [n = 5], Enterobacter cloacae [n = 5], Klebsiella pneumoniae [n = 3] and Klebsiella aerogenes [n = 1]) were identified. There were diverse PFGE profiles and sequence types (STs) in most of the species except for K. oxytoca. All isolates of K. oxytoca belonged to ST29 with similar PFGE profiles, suggesting their clonal transmission. Plasmid analysis by WGS revealed that all 22 isolates but one shared a ca. 50-kb IncN plasmid backbone with blaIMP-6 suggesting interspecies gene transmission, and typing of plasmids explained epidemiological links among cases. A case-control study showed pancreatoduodenectomy, changing drains in fluoroscopy room, continuous peritoneal lavage and enteric fistula were associated with IMP-6-CPE acquisition among the patients. Plasmid analysis of isolates in an outbreak of IMP-6-CPE suggested interspecies gene transmission and helped to clarify hidden epidemiological links between cases.

DOI： 10.1038/s41598-020-60659-2

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:p>
<jats:named-content content-type="genus-species">Helicobacter suis</jats:named-content>
strain SNTW101c, which was originally obtained from a patient with nodular gastritis, has been maintained in mouse stomach because of difficulty culturing it
<jats:italic>in vitro</jats:italic>
. Recently, we succeeded in culturing this strain
<jats:italic>in vitro</jats:italic>
. Here, we report the complete genome sequence of
<jats:named-content content-type="genus-species">H. suis</jats:named-content>
strain SNTW101c.
</jats:p>

DOI： 10.1128/MRA.01340-19

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Cellular and Infection Microbiology  

Nationwide increases in Mycoplasma pneumoniae pneumonia cases in Japan were reported in 2011, 2012, 2015, and 2016. In this study, we isolated 554 M. pneumoniae strains in 4 areas in Japan (Kanagawa, Okayama, Osaka, and Saitama) between 2006 and 2019, and performed genotyping analysis. More than 80% of the strains isolated in 2011 and 2012 harbored type 1 p1 adhesin gene; however, strains harboring type 2 or its variant p1 gene increased in 2015 and 2016 and dominated after 2017. These findings suggested that a shift in the prevalent genotype of M. pneumoniae clinical strains occurred recently in Japan. More than 90% of the type 1 strains isolated after 2010 harbored macrolide-resistance mutations in their 23S rRNA gene, whereas most type 2 lineage strains had no such mutations. Consequently, the increase in type 2 lineage strains in Japan has reduced the macrolide resistance rate of clinical M. pneumoniae strains. During this analysis, we also identified M. pneumoniae strains carrying a novel variant type 1 p1 gene, and we classified it as type 1b. We then sequenced the genomes of 81 selected M. pneumoniae strains that we collected between 1976 and 2017 in Japan, and compared them with 156 M. pneumoniae genomes deposited in public databases to provide insights into the interpretation of M. pneumoniae genotyping methods, including p1 typing, multiple-locus variable-number tandem repeat analysis (MLVA), multi-locus sequence typing (MLST), and typing by 8 single-nucleotide polymorphism markers (SNP-8). As expected, p1 typing, MLST, and SNP-8 results exhibited good correlation with whole-genome SNP analysis results in terms of phylogenetic relationships; however, MLVA typing results were less comparable to those of the other methods. MLVA may be useful for the discrimination of strains derived from a single outbreak within a limited area; however, is not reliable for classification of strains collected from distantly separated areas at different time points. This study showed the usefulness of genome-based comparison of M. pneumoniae for molecular epidemiology. Genome sequencing of more strains will improve our understanding of global propagation routes of this pathogen and evolutionary aspects of M. pneumoniae strains.

DOI： 10.3389/fcimb.2020.00385

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Microbiology  

Epigenetic DNA base methylation plays important roles in gene expression regulation. We here describe a gene expression regulation network consisting of many DNA methyltransferases each frequently changing its target sequence-specificity. Our object Helicobacter pylori, a bacterium responsible for most incidence of stomach cancer, carries a large and variable repertoire of sequence-specific DNA methyltransferases. By creating a dozen of single-gene knockout strains for the methyltransferases, we revealed that they form a network controlling methylome, transcriptome and adaptive phenotype sets. The methyltransferases interact with each other in a hierarchical way, sometimes regulated positively by one methyltransferase but negatively with another. Motility, oxidative stress tolerance and DNA damage repair are likewise regulated by multiple methyltransferases. Their regulation sometimes involves translation start and stop codons suggesting coupling of methylation, transcription and translation. The methyltransferases frequently change their sequence-specificity through gene conversion of their target recognition domain and switch their target sets to remodel the network. The emerging picture of a metamorphosing gene regulation network, or firework, consisting of epigenetic systems ever-changing their specificity in search for adaptation, provides a new paradigm in understanding global gene regulation and adaptive evolution.

DOI： 10.3389/fmicb.2020.01628

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Objectives To examine the association between penicillin susceptibility of Streptococcus pneumoniae and penicillin consumption in Japan. Methods We used Japan Nosocomial Infection Surveillance data on the susceptibility of S. pneumoniae and sales data obtained from IQVIA Services Japan K.K. for penicillin consumption. We analysed both sets of data by decomposing them into seasonality and chronological trend components. The cross-correlation function was checked using Spearman's rank correlation coefficient to examine the correlation between susceptibility and consumption. Results After adjusting for seasonality, the susceptibility of S. pneumoniae to penicillins gradually improved (55.7% in 2013 and 60.6% in 2018, respectively) and penicillin consumption increased during the same period (0.76 defined daily doses per 1,000 inhabitants per day [DID] in 2013, and 0.89 DID in 2018). The results showed positive cross-correlation (coefficient 0.801, p-value < 0.001). In contrast, cephalosporin consumption decreased (3.91 DID in 2013 and 3.19 DID in 2018) and showed negative cross-correlation with susceptibility of S. pneumoniae to penicillins (coefficient -0.981, p-value < 0.001). Conclusions The rates of penicillin-susceptible S. pneumoniae isolates did not negatively correlate with penicillin consumption at the population level. Increased penicillin consumption might not impair the penicillin susceptibility of S. pneumoniae.

DOI： 10.1371/journal.pone.0240655

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Enterobacterales resistant to carbapenems, a class of last-resort antimicrobials, are ranked as an "urgent"and "critical"public health hazard by CDC and WHO. IMP-type carbapenemase- containing Enterobacterales are endemic in Japan, and blaIMP-6 is one of the notable carbapenemase genes responsible for the resistance. The gene is plasmidencoded and confers resistance to meropenem, but not to imipenem. Therefore, IMP-6- producing Enterobacterales isolates are occasionally overlooked in clinical laboratories and are referred to as 'stealth-type'. Since previous reports in Japan were confined only to some geographical regions, their distribution across prefectures and the factors affecting the distribution remain unclear. Here, we revealed the dynamics of the geographical distribution of Enterobacterales with IMP-6 phenotype associated with antimicrobial use in Japan. We utilized comprehensive national surveillance data of all routine bacteriological test results from more than 1,400 hospitals in 2015 and 2016 to enumerate Escherichia coli and Klebsiella pneumoniae isolates with the antimicrobial susceptibility pattern (phenotype) characteristic of IMP-6 (imipenem susceptible, meropenem resistant), and to tabulate the frequency of isolates with the phenotype for each prefecture. Isolates were detected in approximately half of all prefectures, and combined analysis with the national data of antimicrobial usage revealed a statistically significant association between the frequency and usage of not carbapenems but third-generation cephalosporins (p = 0.006, logistic mixed-effect regression) and a weaker association between the frequency and usage of fluoroquinolones (p = 0.043). The usage of third-generation cephalosporins and fluoroquinolones may select the strains with the IMP-6 phenotype, and contribute to their occasional spread. We expect the findings will promote antimicrobial stewardship to reduce the spread of the notable carbapenemase gene. Copyright:

DOI： 10.1371/journal.pone.0243630

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Microbiology  

In the original article, there was an error. “HpyPIX” was misspelled as “HpyPX” throughout the article. The correct spelling is “HpyPIX”. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated..

DOI： 10.3389/fmicb.2020.596598

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

A major issue in the surveillance of antimicrobial resistance (AMR) is “de-duplication” or removal of repeated isolates, for which there exist multiple methods. The World Health Organization (WHO) Global Antimicrobial Resistance Surveillance System (GLASS) requires de-duplication by selecting only the first isolate of a given bacterial species per patient per surveillance period per specimen type per age group, gender, and infection origin stratification. However, no study on the comparative application of this method has been reported. The objective of this study was to evaluate differences in data tabulation between the WHO GLASS and the Japan Nosocomial Infections Surveillance (JANIS) system, which counts both patients and isolates after removing repeated isolates of the same bacterial species isolated from a patient within 30 days, regardless of specimen type, but distinguishing isolates with change of antimicrobial resistance phenotype. All bacterial data, consisting of approximately 8 million samples from 1795 Japanese hospitals in 2017 were exported from the JANIS database, and were tabulated using either the de-duplication algorithm of GLASS, or JANIS. We compared the tabulated results of the total number of patients whose blood and urine cultures were taken and of the percentage of resistant isolates of Escherichia coli for each priority antibiotic. The number of patients per specimen type tabulated by the JANIS method was always smaller than that of GLASS. There was a small (< 3%) difference in the percentage of resistance of E. coli for any antibiotic between the two methods in both out- and inpatient settings and blood and urine isolates. The two tabulation methods did not show considerable differences in terms of the tabulated percentages of resistance for E. coli. We further discuss how the use of GLASS tabulations to create a public software and website that could help to facilitate the understanding of and treatment against AMR.

DOI： 10.1371/journal.pone.0228234

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier BV  

DOI： 10.2139/ssrn.3663972

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

Purpose. Human-adapted Bordetella parapertussis is one of the causative agents of whooping cough; however, there are currently no genotyping systems with high discriminatory power for this bacterial pathogen. We therefore aimed to develop a multilocus variable-number tandem repeat analysis (MLVA) for human-adapted B. parapertussis. Methodology. Four highly polymorphic variable number tandem repeat (VNTR) loci in the B. parapertussis genome were selected and amplified by multiplex PCR. MLVA was performed based on the number of tandem repeats at VNTR loci. The discriminatory power of MLVA was evaluated with three laboratory reference strains and 50 human isolates of B. parapertussis. Results. Multiplex PCR-based MLVA characterized 53 B. parapertussis reference strains and isolates into 25 MLVA types and the Simpson diversity index was 0.91 (95 % confidence interval, 0.86–0.97). The three reference strains exhibited different MLVA types. Thirty-one Japanese isolates, ten French isolates and three Taiwanese isolates belonged to fourteen, nine and three MLVA types, respectively. In contrast, all five Australian isolates belonged to the same type. Two Japanese isolates collected from patients with known epidemiological links had the same type. Conclusion. Our novel MLVA method has high discriminatory power for genotyping human B. parapertussis. Regarding this organism, this genotyping system is a promising tool for epidemiological surveillance and investigating outbreaks.

DOI： 10.1099/jmm.0.001095

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Macromolecular Bioscience  

Vaccines typically contain an antigen, delivery system (vehicle), and adjuvant, all of which contribute to inducing a potent immune response. Consequently, design of new vaccines is difficult, because the contributions and interactions of these components are difficult to distinguish. Here, it is aimed to develop an easy-to-use, non-immunogenic, injectable depot system for sustained antigen release that will be suitable for assessing the efficacy of prolonged antigen exposure per se for inducing an immune response. This should mimic real-life infections. Recombinant elastin-like polypeptides with periodic cysteine residues (cELPs) are selected, which reportedly show little or no immunogenicity, as carriers and tetanus toxoid (Ttd) as an antigen. After subcutaneous injection of the mixture, cELP rapidly forms a disulfide cross-linked hydrogel in situ, within which Ttd is physically incorporated, affording a biodegradable antigen depot. A series of Ttd-containing hydrogels is examined. A single injection induces high levels of tetanus antibody with high avidity for at least 20 weeks in mice. The chain length of cELP proves critical, whereas differences in hydrophobicity has little effect, although hydrophilic cELPs are more rapidly biodegraded. This system's ability to distinguish the contribution of sustained antigen release to antibody induction should be helpful for rational design of next-generation vaccines.

DOI： 10.1002/mabi.201900167

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

Leprosy, an important infectious disease in humans caused by Mycobacterium leprae (Mle), remains endemic in many countries. Notably, the pathogen cannot be cultured in vitro, except in mouse footpads in vivo. The molecular basis of these characteristics and the mechanisms remain unknown. Consequently, analysis of Mle growth and survival is urgently needed to develop novel therapies against leprosy, including rapid, simple, and specific methods to detect infection. Here, we demonstrated the functional role and contribution of Mle-DNA gyrase, which regulates DNA topology, DNA replication, and chromosome segregation to promote bacterial growth and survival, in Mle growth and survival in vitro and in vivo. The optimum temperature for Mle-DNA gyrase activity was 30 °C. When the DNA gyrB-gyrA genes in Mycobacterium smegmatis were replaced with the Mle gyrase genes by allelic exchange, the recombinants could not grow at 37 °C. Moreover, using radiorespirometry analysis for viability of Mle bacilli, we found that Mle growth was more vigorous at 25–30 °C than at 37 °C, but was inhibited above 40 °C. These results propose that DNA gyrase is a crucial factor for Mle growth and survival and its sensitivity to temperature may be exploited in heat-based treatment of leprosy.

DOI： 10.1038/s41598-019-47364-5

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

The genus Acinetobacter comprises many species that can cause infectious diseases. Despite their importance as nosocomial pathogens, the clinical distributions of individual species or clones are not well understood. In this study, we aimed to characterize 13 Acinetobacter strains isolated from blood cultures from Osaka City University Hospital. We conducted whole-genome sequencing to reveal their genetic background. We also performed PCR-based open reading frame typing (POT) and compared the results with those of multilocus sequence typing (MLST) to confirm its reliability as a genotyping method. Although biochemical analysis suggested that most isolates were A. baumannii, genomic analysis revealed that the collection of Acinetobacter isolates comprised six different species, with non-baumannii Acinetobacter species representing the majority. All strains possessed an inherent ADC-type β-lactamase gene, whereas the distribution of OXA-type enzymes was limited to A. baumannii, A. pittii, and A. colistiniresistens. While MLST properly discriminated four A. baumannii strains as different clones, POT failed to distinguish three of the four A. baumannii strains from each other, highlighting a potential pitfall that may be encountered when applying POT to 263718699pidemiological A. baumannii strains.

DOI： 10.7883/yoken.JJID.2018.403

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Emerging Infectious Diseases  

Bacillus cereus is associated with foodborne illnesses characterized by vomiting and diarrhea. Although some B. cereus strains that cause severe extraintestinal infections and nosocomial infections are recognized as serious public health threats in healthcare settings, the genetic backgrounds of B. cereus strains causing such infections remain unknown. By conducting pulsed-field gel electrophoresis and multilocus sequence typing, we found that a novel sequence type (ST), newly registered as ST1420, was the dominant ST isolated from the cases of nosocomial infections that occurred in 3 locations in Japan in 2006, 2013, and 2016. Phylogenetic analysis showed that ST1420 strains belonged to the Cereus III lineage, which is much closer to the Anthracis lineage than to other Cereus lineages. Our results suggest that ST1420 is a prevalent ST in B. cereus strains that have caused recent nosocomial infections in Japan.

DOI： 10.3201/eid2505.171890

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Global Antimicrobial Resistance  

DOI： 10.1016/j.jgar.2019.02.004

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Global Antimicrobial Resistance  

Objectives: Plasmids harbouring antimicrobial resistance determinants in clinical strains are a significant public-health concern worldwide. The present study investigated such plasmids in clinical isolates of Shigella flexneri. Methods: A total of 162 Shigella isolates were obtained from stool specimens in the year 2015. Among the 70 multidrug-resistant (MDR) Shigella spp., 27 S. flexneri isolates were randomly selected for further characterisation. Antimicrobial resistance genes (ARGs) and plasmid incompatibility (Inc) types were analysed. Results: IncFII plasmids were found in 63% (17/27) of the studied S. flexneri isolates. ARGs such as dhfr1a (81%), sulII (74%), bla OXA (74%), bla TEM (33%), bla AmpC (30%), qnrS (15%) and qnrB (4%) were identified by PCR, whereas bla CTX-M was not detected. Next-generation sequencing of a representative S. flexneri IncFII-type plasmid (pSF470) revealed the presence of bla TEM1-B , bla DHA-1 , qnrB10, mphA, sulI, sulII, strA, strB and tetR ARGs along with the intI1 integrase gene. In addition, pMLST analysis showed that the replicon belonged to F2:A-:B- type. Conclusions: This study helps to know the prevalent plasmid types in MDR Shigella isolates and will improve our understanding of resistance dissemination among enteric bacteria. ARGs in plasmids further highlight the importance of such studies in enteric bacteria.

DOI： 10.1016/j.jgar.2018.10.014

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記述言語：英語   掲載種別：研究論文（国際会議プロシーディングス）   出版者・発行元：ELSEVIER SCI LTD  

DOI： 10.1016/j.ijid.2018.11.131

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Biochemical and Biophysical Research Communications  

Mycoplasma pneumoniae forms an attachment organelle at one cell pole, binds to the host cell surface, and glides via a unique mechanism. A 170-kDa protein, P1 adhesin, present on the organelle surface plays a critical role in the binding and gliding process. In this study, we obtained a recombinant P1 adhesin comprising 1476 amino acid residues, excluding the C-terminal domain of 109 amino acids that carried the transmembrane segment, that were fused to additional 17 amino acid residues carrying a hexa-histidine (6 × His) tag using an Escherichia coli expression system. The recombinant protein showed solubility, and chirality in circular dichroism (CD). The results of analytical gel filtration, ultracentrifugation, negative-staining electron microscopy, and small-angle X-ray scattering (SAXS) showed that the recombinant protein exists in a monomeric form with a uniformly folded structure. SAXS analysis suggested the presence of a compact and ellipsoidal structure rather than random or molten globule-like conformation. Structure model based on SAXS results fitted well with the corresponding structure obtained with cryo-electron tomography from a closely related species, M. genitalium. This recombinant protein may be useful for structural and functional studies as well as for the preparation of antibodies for medical applications.

DOI： 10.1016/j.bbrc.2018.11.132

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

We characterized 419 Mycoplasma pneumoniae isolates collected between 2011 and 2017 in Osaka prefecture of Japan. This analysis revealed high prevalence of macrolide-resistant M. pneumoniae (MRMP) in Osaka during 2011 and 2014 with annual detection rates of MRMP strains between 71.4% and 81.8%. However, in 2015 and after, the detection rate of MRMP decreased significantly and did not exceed 50%. Genotyping of the p1 gene of these isolates showed that most of MRMP strains harbored type 1 p1 gene. In contrast, strains expressing p1 gene type 2 or its variant were largely macrolide-susceptible M. pneumoniae (MSMP) strains. There was a strong correlation between p1 gene genotype and the presence of mutations conferring macrolide resistance in M. pneumoniae isolated in Osaka. These results indicate that lower incidence of MRMP strains in Osaka from 2015 was associated with the relative increase of p1 gene type 2 lineage strains. During these experiments, we also isolated three M. pneumoniae strains that showed irregular typing pattern in the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the p1 gene. Two of these strains harbored new variants of type 2 p1 gene and were designated as type 2f and 2g. The remaining strain with an irregular typing pattern had a large deletion in the p1 operon.

DOI： 10.1371/journal.pone.0209938

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Antimicrobial resistance and infection control  

Background: To clarify the molecular epidemiology of carbapenem-resistant Enterobacter cloacae complex (CREC) and the risk factors for acquisition of carbapenemase-producing E. cloacae complex (CPEC). Methods: Using clinical CREC isolates detected in a Japanese university hospital over 4 years, carbapenemase production was screened with phenotypic methods. Carbapenemase genes were analysed by PCR and sequencing. Molecular epidemiological analyses were conducted with repetitive extragenic palindromic (REP)-PCR and multilocus sequence typing (MLST). CRECs were identified to the subspecies level by hsp60 sequencing. Whole-genome sequencing of plasmids was conducted. A case-control study was performed to identify risk factors for acquisition of CPEC among patients with CREC. Results: Thirty-nine CRECs including 20 CPECs carrying blaIMP-1 were identified. Patients with CPEC had longer hospital stay before detection (26.5 days vs. 12 days, p = 0.008), a urinary catheter (odds ratio [OR], 5.36; 95% confidence interval [CI], 1.14-30.9; p = 0.023), or intubation (OR, 7.53; 95% CI, 1.47-53.8; p = 0.008) compared to patients without CPEC. Four genetically closely related CPEC clusters were observed, which showed that three of four CPEC clusters corresponded to E. asburiae (ST 53), E. hormaechei subsp. steigerwaltii (ST 113 and ST 1047) and E. cloacae subsp. cloacae (ST 513) by MLST and hsp60 sequencing. Seven representative plasmids shared structures with class I integron containing blaIMP-1 and IncHI2A replicon type. Conclusions: A longer hospital stay, presence of a urinary catheter, and intubation are risk factors for CPEC acquisition. In addition to horizontal transmission of genetically indistinguishable CPECs, IncHI2A plasmid carrying blaIMP-1 appeared to be transferred among genetically different ECs.

DOI： 10.1186/s13756-019-0578-3

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Pertussis is a human respiratory infection caused by the gram-negative bacterium, Bordetella pertussis. To evaluate the pertussis burden and vaccine efficacy, diagnosis and epidemiological surveillance should be based on accurate and valid diagnostic methods. Recently, the serodiagnostic tests Novagnost Bordetella pertussis IgA and IgM were approved in Japan for pertussis diagnostics. Although the anti-pertussis toxin (PT) IgG assay has been used for pertussis diagnosis worldwide, little is known about the anti-B. pertussis IgA and IgM assays. In this study, serum samples from 460 healthy donors were examined to determine the seroprevalence of anti-B. pertussis IgA and IgM in a Japanese population, and its correlation with donor age. Our data demonstrated that anti-B. pertussis IgA and IgM are positively and negatively correlated with age (r = 0.27, r = -0.37; P < 0.001, respectively). Age-specific analysis revealed high titers of anti-B. pertussis IgA in adults (46–50 years), while anti-B. pertussis IgM titers were high in schoolchildren (6–10, 11–15 years). When applying the arbitrary cut-off values for these ages, 17.6% and 39.5% of healthy donors were interpreted as pertussis-positive or indeterminate with anti-B. pertussis IgA (46–50 years) and IgM (11–15 years) titers, respectively. Overall, our findings indicated that the Novagnost Bordetella pertussis IgA and IgM testing could be greatly affected by subject age, limiting its value for pertussis diagnosis.

DOI： 10.1371/journal.pone.0219255

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbial Pathogenesis  

Although standard antibiotic therapy is performed for diarrhea and pseudomembranous colitis caused by Clostridium difficile, a high recurrence rate of C. difficile infection (CDI) remains a major problem. We previously showed that a membrane fraction of nontoxigenic C. difficile (ntCDMF) was effective as a vaccine antigen by in vitro experiments. In this study, we examined whether ntCDMF had an in vivo effect in animal challenge experiments. By intrarectal immunization with ntCDMF, the number of C. difficile cells in feces of mice was decreased approximately 99% compared to the control mice. In addition, survival rate of C. difficile-challenged hamsters was increased almost 30% by immunization with ntCDMF. These results showed that ntCDMF could be a practical vaccine candidate.

DOI： 10.1016/j.micpath.2018.06.039

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Journal of Infection Control  

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is the most common antimicrobial-resistant organism identified in Japanese health care facilities. This study analyzed the clinical and economic burdens attributable to methicillin resistance in S aureus in Japanese hospitals. Methods: We retrospectively investigated data from 14,905 inpatients of 57 hospitals combined with data from nosocomial infection surveillance and administrative claim databases. The participants were inpatients with admission from April 1, 2014, to discharge on March 31, 2016. The outcomes were evaluated according to length of stay, hospital charges, and in-hospital mortality. We compared the disease burden of MRSA infections with methicillin-susceptible S aureus (MSSA) infections based on patients’ characteristics and onset periods. Results: We categorized 7,188 and 7,717 patients into MRSA and MSSA groups, respectively. The adjusted effects of the MRSA group were 1.03-fold (95% confidence interval [CI] 1.01-1.05) and 1.04-fold (95% CI, 1.01-1.06), respectively, with an odds ratio of 1.14 (95% CI, 1.02-1.27). Conclusions: The results of this study found that patient severity and onset delays were positively associated with both MRSA and burden and that the effect of methicillin resistance remained significant after adjustment.

DOI： 10.1016/j.ajic.2018.04.214

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Vaccine  

Background: Haemophilus influenzae type b (Hib) vaccine was introduced as a voluntary vaccine in December 2008 and was included in the national routine immunization program in April 2013 in Japan. Currently, no nationwide data are available to evaluate the effectiveness of Hib vaccine in Japan. Methods: To evaluate the effectiveness of Hib vaccine in Japan, nationwide active population-based surveillance of culture-proven invasive infections caused by H. influenzae in children was performed in 2008–2017 in 10 prefectures in Japan (covering approximately 23% of the total Japanese population). Clinical data were recorded on a standardized case report form. Capsular type and antimicrobial susceptibility of the H. influenzae isolates were examined. The incidence rate ratio (IRR) and its confidence interval (CI) were calculated to compare data from 5 years before and that from after the introduction of the national routine Hib vaccine immunization program. Results: During the 10-year study period, 566 invasive H. influenzae disease cases including 336 meningitis cases were identified. The average number of invasive H. influenzae disease cases among children <5 years of age during 2013–2017 decreased by 93% (IRR: 0.07, 95%CI 0.05–0.10, p < 0.001) compared with those occurring during 2008–2012. Hib strains have not been isolated from invasive H. influenzae disease cases since 2014; however, non-typeable H. influenzae and H. influenzae type f isolates have been noted as causes of invasive H. influenzae diseases among children <5 years in the post-Hib vaccine era. Conclusions: After the governmental subsidization of the Hib vaccine, invasive Hib disease cases decreased dramatically in the study population, as per our surveillance. Continuous surveillance is necessary to monitor the effectiveness of Hib vaccine and for detecting any emerging invasive capsular types.

DOI： 10.1016/j.vaccine.2018.08.029

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:p>
<jats:named-content content-type="genus-species">Haemophilus influenzae</jats:named-content>
type b (Hib) conjugate vaccines have led to dramatic reductions in Hib disease among young children worldwide. Nontypeable
<jats:named-content content-type="genus-species">H. influenzae</jats:named-content>
(NTHi) is now the major cause of invasive
<jats:named-content content-type="genus-species">H. influenzae</jats:named-content>
infections.
</jats:p>

DOI： 10.1128/JCM.00141-18

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

Frequent use of broad-spectrum antimicrobial classes has been reported in Japan; however, little is known about the long-term trend of national antimicrobial consumption, and that of individual agents. This study analyzed the national sales data of systemic antimicrobials from 2004 to 2016, derived from the IMS Japan Pharmaceutical Market database, to assess the consumption patterns of antimicrobial classes and agents in Japan. The number of defined daily doses per 1000 inhabitants per day (DID) was calculated for each antimicrobial agent. During the last 13 years, total antimicrobial consumption fluctuated by only 5% around the average of 14.41 DID. In 2016, the most used class was macrolides (32%), followed by cephalosporins (28%) and fluoroquinolones (19%). Oral agents comprised a large proportion (93%) of antimicrobial consumption. The most used agent, clarithromycin, accounted for 25% of all oral compounds used in 2016. The consumption of oral agents with high bioavailability, such as fluoroquinolones, amoxicillin, and sulfamethoxazole/trimethoprim increased, whereas that of cephalosporins decreased. In 2016, ceftriaxone was the most consumed parenteral agent, followed by cefazolin. The consumption of parenteral agents increased after 2009 when high-dose regimens of piperacillin/tazobactam, meropenem, and ampicillin/sulbactam were approved by the health insurance system. National antimicrobial consumption has been stable over the last 13 years. Moreover, shifts in the use of agents with high bioavailability and those approved for high-dose regimens were observed. However, the increased use of broad-spectrum agents is worrisome. A multifaceted approach is required to reduce overall antimicrobial consumption.

DOI： 10.1016/j.jiac.2018.01.003

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
<jats:named-content content-type="genus-species">Mycobacterium marinum</jats:named-content>
is a slowly growing, broad-host-range mycobacterial species. Here, we report the complete genome sequence of a
<jats:named-content content-type="genus-species">Mycobacterium marinum</jats:named-content>
type strain that was isolated from tubercles of diseased fish. This sequence will provide essential information for future taxonomic and comparative genome studies of its relatives.
</jats:p>

DOI： 10.1128/genomeA.00397-18

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

Purpose. The decline in methicillin-resistant Staphylococcus aureus (MRSA) isolation rates has become a general observation worldwide, including Japan. We hypothesized that some genetic shift in MRSA might cause this phenomenon, and therefore we investigated the genetic profiles among MRSA clinical isolates obtained from three different epidemic phases in Japan. Methodology. A total of 353 MRSA isolates were selected from 202 medical facilities in 1990 (pre-epidemic phase), 2004 (epidemic phase) and 2016 (post-epidemic phase). Molecular typing was performed by PCR detection of 22 genes using the polymerase chain reaction (PCR)-based ORF typing (POT) system, including an additional eight genes including small genomic islets and seven toxin genes. Results. Isolates with a POT1 of score 93, identified as presumed clonal complex (pCC)5-staphylococcal cassette chromosome mec (SCCmec) type II including ST5-SCCmec type II New York/Japan clone, represented the major epidemic MRSA lineage in 1990 and 2004. In 2016, however, a marked decrease in isolates with a POT1 score of 93, along with changes in the epidemiology of toxin genes carried, was noted, where the carriers of tst genes including the tst-sec combination were markedly reduced, and those possessing the seb gene alone were markedly increased. Rather, isolates with a POT1 score of 106, including pCC1 or pCC8 among the isolates with SCCmec type IV, which often links to communityassociated MRSA, were predominant. Interestingly, the pCC1 and pCC8 lineages were related to sea and tst-sec carriage, respectively. Conclusions. Over time, a transition in MRSA genetic profiles from a POT1 score of 93 in 1990 and 2004 to 106 in 2014 was found in Japan.

DOI： 10.1099/jmm.0.000687

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Vancomycin-intermediately resistant Staphylococcus aureus (VISA) and heterogeneous VISA (hVISA) are associated with treatment failure. hVISA contains only a subpopulation of cells with increased minimal inhibitory concentrations, and its detection is problematic because it is classified as vancomycin-susceptible by standard susceptibility testing and the gold-standard method for its detection is impractical in clinical microbiology laboratories. Recently, a research group developed a machine-learning classifier to distinguish VISA and hVISA from vancomycin-susceptible S. aureus (VSSA) according to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) data. Nonetheless, the sensitivity of hVISA classification was found to be 76%, and the program was not completely automated with a graphical user interface. Here, we developed a more accurate machine-learning classifier for discrimination of hVISA from VSSA and VISA among MRSA isolates in Japanese hospitals by means of MALDI-TOF MS data. The classifier showed 99% sensitivity of hVISA classification. Furthermore, we clarified the procedures for preparing samples and obtaining MALDI-TOF MS data and developed all-in-one software, hVISA Classifier, with a graphical user interface that automates the classification and is easy for medical workers to use; it is publicly available at https://github.com/bioprojects/hVISAclassifier. This system is useful and practical for screening MRSA isolates for the hVISA phenotype in clinical microbiology laboratories and thus should improve treatment of MRSA infections.

DOI： 10.1371/journal.pone.0194212

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
<jats:italic>Mycobacterium abscessus</jats:italic>
subsp.
<jats:italic>bolletii</jats:italic>
is a rapidly growing mycobacterial organism for which the taxonomy is unclear. Here, we report the complete genome sequence of a
<jats:italic>Mycobacterium abscessus</jats:italic>
subsp.
<jats:italic>bolletii</jats:italic>
type strain. This sequence will provide essential information for future taxonomic and comparative genome studies of these mycobacteria.
</jats:p>

DOI： 10.1128/genomeA.01530-17

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
Multidrug-resistant (MDR)
<jats:named-content content-type="genus-species">Acinetobacter</jats:named-content>
spp. have been globally disseminated in association with the successful clonal lineage
<jats:named-content content-type="genus-species">Acinetobacter baumannii</jats:named-content>
international clone II (IC II). Because the prevalence of MDR
<jats:named-content content-type="genus-species">Acinetobacter</jats:named-content>
spp. in Japan remains very low, we characterized all
<jats:named-content content-type="genus-species">Acinetobacter</jats:named-content>
spp. (
<jats:italic>n</jats:italic>
= 866) from 76 hospitals between October 2012 and March 2013 to describe the entire molecular epidemiology of
<jats:named-content content-type="genus-species">Acinetobacter</jats:named-content>
spp. The most prevalent species was
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
(
<jats:italic>n</jats:italic>
= 645; 74.5%), with
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
IC II (
<jats:italic>n</jats:italic>
= 245) accounting for 28.3% of the total. Meropenem-resistant isolates accounted for 2.0% (
<jats:italic>n</jats:italic>
= 17) and carried IS
<jats:italic>Aba1-bla</jats:italic>
<jats:sub>OXA-23-like</jats:sub>
(
<jats:italic>n</jats:italic>
= 10),
<jats:italic>bla</jats:italic>
<jats:sub>IMP</jats:sub>
(
<jats:italic>n</jats:italic>
= 4), or IS
<jats:italic>Aba1-bla</jats:italic>
<jats:sub>OXA-51-like</jats:sub>
(
<jats:italic>n</jats:italic>
= 3). Multilocus sequence typing of 110 representative
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
isolates revealed the considerable prevalence of domestic sequence types (STs).
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
IC II isolates were divided into the domestic sequence type 469 (ST469) (
<jats:italic>n</jats:italic>
= 18) and the globally disseminated STs ST208 (
<jats:italic>n</jats:italic>
= 14) and ST219 (
<jats:italic>n</jats:italic>
= 4). ST469 isolates were susceptible to more antimicrobial agents, while ST208 and ST219 overproduced the intrinsic AmpC β-lactamase.
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
IC II and some
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
non-IC II STs (e.g., ST149 and ST246) were associated with fluoroquinolone resistance. This study revealed that carbapenem-susceptible
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
IC II was moderately disseminated in Japan. The low prevalence of acquired carbapenemase genes and presence of domestic STs could contribute to the low prevalence of MDR
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
. A similar epidemiology might have appeared before the global dissemination of MDR epidemic lineages. In addition, fluoroquinolone resistance associated with
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
IC II may provide insight into the significance of
<jats:named-content content-type="genus-species">A. baumannii</jats:named-content>
epidemic clones.
</jats:p>

DOI： 10.1128/AAC.02190-17

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
β-Lactams are often used to treat
<jats:named-content content-type="genus-species">Helicobacter cinaedi</jats:named-content>
infections; however, the mechanism underlying β-lactam resistance is unknown. In this study, we investigated β-lactam resistance in an
<jats:named-content content-type="genus-species">H. cinaedi</jats:named-content>
strain, MRY12-0051 (MICs of amoxicillin [AMX] and ceftriaxone [CRO], 32 and 128 μg/ml; obtained from human feces). Based on a comparative whole-genome analysis of MRY12-0051 and the CRO-susceptible
<jats:named-content content-type="genus-species">H. cinaedi</jats:named-content>
strain MRY08-1234 (MICs of AMX and CRO, 1 and 4 μg/ml; obtained from human blood), we identified five mutations in genes encoding penicillin-binding proteins (PBPs), including two in
<jats:italic>pbpA</jats:italic>
, one in
<jats:italic>pbp2</jats:italic>
, and two in
<jats:italic>ftsI</jats:italic>
. Transformation and penicillin binding assays indicated that CRO resistance was mainly associated with mutations in
<jats:italic>pbpA</jats:italic>
; mutations in
<jats:italic>ftsI</jats:italic>
also led to increased resistance to AMX. Knocking out
<jats:italic>cmeB</jats:italic>
and
<jats:italic>cmeD</jats:italic>
, which encode resistance-nodulation-division-type efflux pump components, in
<jats:named-content content-type="genus-species">H. cinaedi</jats:named-content>
type strain CCUG18818 (AMX MIC, 4 to 8 μg/ml) resulted in 8- and 64-fold decreases, respectively, in the AMX MIC. Hence, MICs of AMX in
<jats:named-content content-type="genus-species">H. cinaedi</jats:named-content>
become similar to those of
<jats:named-content content-type="genus-species">Helicobacter pylori</jats:named-content>
isolates in the absence of
<jats:italic>cmeD</jats:italic>
. In conclusion, the difference in susceptibility to β-lactams between
<jats:named-content content-type="genus-species">H. pylori</jats:named-content>
and
<jats:named-content content-type="genus-species">H. cinaedi</jats:named-content>
is explained by differences in efflux pump components. Mutations in
<jats:italic>pbpA</jats:italic>
are the primary determinant of high resistance to β-lactams in
<jats:named-content content-type="genus-species">H. cinaedi</jats:named-content>
.
</jats:p>

DOI： 10.1128/AAC.02036-17

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Carbapenemase-producing Enterobacteriaceae (CPE) are a global concern because these bacteria are resistant to almost all β-lactams. Horizontal interspecies gene transfer via plasmid conjugation has increased the global dissemination of CPE. Recently, an Enterobacteriaceae strain positive for carbapenemase gene but showing a carbapenem-susceptible phenotype was identified, suggesting that these susceptible strains may be challenging to detect solely via antimicrobial susceptibility tests without molecular analysis. Here, we isolated a blaIMP-6 carbapenemase-gene positive but imipenem- and meropenem-susceptible Escherichia coli (ISMS-E) strain A56-1S (imipenem and meropenem minimum inhibitory concentration, 0.125 mg/L), from a human urine specimen in Japan. A56-1S was carbapenemase negative by the Carba NP test, suggesting that IMP-6 production was low or undetectable. Thus, to characterize the mechanism of this phenotype, a meropenem-resistant E. coli A56-1R strain was obtained using meropenem-selection. A56-1R was positive for carbapenemase production by the Carba NP test, and blaIMP-6 transcription in A56-1R was 53-fold higher than in A56-1S, indicating that blaIMP-6 in A56-1S is negatively regulated at the transcriptional level. Comparative genomic analysis between the two strains revealed that the alleviation of restriction of DNA (ardK) gene encoding a putative transcription factor is disrupted by the IS26 insertion in A56-1R. A cotransformation assay of ardK and the regulatory element upstream of blaIMP-6 showed repression of blaIMP-6 transcription, indicating that ArdK negatively modulates blaIMP-6 transcription. ArdK binding and affinity assays demonstrated that ArdK directly binds to the regulatory element upstream of blaIMP-6 with dissociation constant values comparable to those of general transcription factors. The IMP-6 carbapenemase showed low hydrolytic activity against imipenem, resulting in an imipenem-susceptible and meropenem-resistant (ISMR) phenotype (previously reported as a stealth phenotype). However, the low expression of IMP-6 in the A56-1S strain could be a typical characteristic of ISMS-E due to gene repression, indicating that conventional antimicrobial susceptibility tests might be unable to detect such strains even when using both imipenem and meropenem. Bacteria that exhibit the ISMS phenotype could play a potential role as undetectable reservoirs and might facilitate gene transfer to other organisms while avoiding detection.

DOI： 10.1371/journal.pone.0208976

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
<jats:italic>Mycobacterium</jats:italic>
sp. strain shizuoka-1 is a rapidly growing scotochromogenic mycobacterium and was isolated from well water for a bathing facility in Shizuoka Prefecture in Japan. Here, we report the draft sequence of its genome, comprising a 6.5-Mb chromosome. This mycobacterium has 83.1% identity with
<jats:named-content content-type="genus-species">Mycobacterium rhodesiae</jats:named-content>
, a human pathogen.
</jats:p>

DOI： 10.1128/genomeA.01309-17

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

This is the first report on large-scale experimental production of an equine antivenom against the redback spider (Latrodectus hasseltii) lived in Japan. We captured 10,000 redback spiders in Japan and prepared the toxoids of crude venom extract, mixed the toxoids with a mineral oil adjuvant, and immunized healthy horses repeatedly over a period of several weeks. Thereafter, we separated the horse plasma, purified the γ-globulin fraction, and stocked it as a purified antivenom concentrate. Consequently, we manufactured approximately 6,500 vials of a single-dose freeze-dried test lot from a portion of the purified γ-globulin fraction, equivalent to the extract derived from 520 spiders. This test lot had an antitoxin titer comparable to that of a similar drug commercially available overseas (a liquid preparation), and the other quality met all quality reference specifications based on the Minimum Requirements for Biological Products and other guidelines relevant to existing antivenom drug products in Japan.

DOI： 10.7883/yoken.JJID.2017.125

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Helicobacter cinaedi is associated with nosocomial infections. The CRISPR-Cas system provides adaptive immunity against foreign genetic elements. We investigated the CRISPR-Cas system in H. cinaedi to assess the potential of the CRISPR-based microevolution of H. cinaedi strains. A genotyping method based on CRISPR spacer organization was carried out using 42 H. cinaedi strains. The results of sequence analysis showed that the H. cinaedi strains used in this study had two CRISPR loci (CRISPR1 and CRISPR2). The lengths of the consensus direct repeat sequences in CRISPR1 and CRISPR2 were both 36 bp-long, and 224 spacers were found in the 42 H. cinaedi strains. Analysis of the organization and sequence similarity of the spacers of the H. cinaedi strains showed that CRISPR arrays could be divided into 7 different genotypes. Each genotype had a different ancestral spacer, and spacer acquisition/deletion events occurred while isolates were spreading. Spacer polymorphisms of conserved arrays across the strains were instrumental for differentiating closely-related strains collected from the same hospital. MLST had little variability, while the CRISPR sequences showed remarkable diversity. Our data revealed the structural features of H. cinaedi CRISPR loci for the first time. CRISPR sequences constitute a valuable basis for genotyping, provide insights into the divergence and relatedness between closely-related strains, and reflect the microevolutionary process of H. cinaedi.

DOI： 10.1371/journal.pone.0186241

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

Methicillin-resistant Staphylococcus aureus (MRSA) bacteremia is one of the commonest and most life-threatening of all infectious diseases. The morbidity and mortality rates associated with MRSA bacteremia are higher than those associated with bacteremia caused by other pathogens. A common guideline in MRSA bacteremia treatment is to confirm bacteremia clearance through additional blood cultures 2–4 days after initial positive cultures and as needed thereafter. However, no study has presented statistical evidence of how and to what extent confirming a negative follow-up blood culture impacts clinical outcome. We present this evidence for the first time, by combining clinical microbiological data of blood cultures and the DPC administrative claims database; both had been systematically accumulated through routine medical care in hospitals. We used electronic medical records to investigate the clinical background and infection source in detail. By analyzing data from a university hospital, we revealed how survival curves change when a negative follow-up blood culture is confirmed. We also demonstrated confirmation of a negative culture is significantly associated with clinical outcomes: there was a more than three-fold increase in mortality risk (after adjusting for clinical background) if a negative blood culture was not confirmed within 14 days of the initial positive blood culture. Although we used data from only one university hospital, our novel approach and results will be a basis for future studies in several hospitals in Japan to provide statistical evidence of the clinical importance of confirming a negative follow-up blood culture in bacteremia patients, including those with MRSA infections.

DOI： 10.1016/j.jiac.2017.07.009

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Infectious Diseases  

Objectives This study sought to determine the genotypes of circulating Bordetella pertussis, the causative agent of pertussis, in Cambodia by direct molecular typing of clinical specimens. Methods DNA extracts from nasopharyngeal swabs obtained from 82 pertussis patients in 2008–2016 were analyzed by multilocus variable-number tandem repeat analysis (MLVA). B. pertussis virulence-associated allelic genes (ptxA, prn, and fim3) and the pertussis toxin promoter ptxP were also investigated by DNA sequence-based typing. Results Forty-four DNA extracts (54%) yielded a complete MLVA profile, and these were sorted into 8 MLVA types (MT18, MT26, MT27, MT29, MT43, MT72, MT95, and MT200). MT27 and MT29, which are common in developed countries, were the predominant strain types (total 73%). The predominant profile of virulence-associated allelic genes was the combination of ptxP3/ptxA1/prn2/fim3A (48%). MT27 strains were detected during the entire study period, whereas MT29 strains were only found in 2014–2016. Conclusions The B. pertussis population in Cambodia, where a whole-cell pertussis vaccine (WCV) has been continuously used, resembled those observed previously in developed countries where acellular pertussis vaccines are used. Circulating B. pertussis strains in Cambodia were distinct from those in other countries using WCVs.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Group B streptococci (GBS; Streptococcus agalactiae) are the leading cause of neonatal invasive diseases and are also important pathogens for elderly adults. Until now, nearly all GBS with reduced penicillin susceptibility (PRGBS) have shown β-hemolytic activity and grow on sheep blood agar. However, we have previously reported three PRGBS clinical isolates harboring a CylK deletion that form small less hemolytic colonies. In this study, we examined the causes of small, less hemolytic colony formation in these clinical isolates. Iso-genic strains were sequenced to identify the mutation related to a small colony size. We identified a 276_277insG nucleic acid insertion in the thiamin pyrophosphokinase (tpk) gene, resulting in premature termination at amino acid 103 in TPK, as a candidate mutation responsible for small colony formation. The recombinant strain Δtpk, which harbored the 276_277insG insertion in the tpk gene, showed small colony formation. The recombinant strain ΔcylK, which harbored the G379T substitution in cylK, showed a reduction in hemolytic activity. The phenotypes of both recombinant strains were complemented by the expression of intact TPK or CylK, respectively. Moreover, the use of Rapid ID 32 API and VITEK MS to identify strains as GBS was evaluated clinical isolates and recombinant strains. VITEK MS, but not Rapid ID 32 API, was able to accurately identify the strains as GBS. In conclusion, we determined that mutations in tpk and cylK caused small colonies and reduced hemolytic activity, respectively, and characterized the clinical isolates in detail.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

In 2013, national serosurveillance detected a high seroprevalence of antibodies to pertussis toxin (PT) from Bordetella pertussis among Japanese adults. Thus, we aimed to determine the cause(s) of this high seroprevalence, and analyzed the titers of antibodies to PT and filamentous hemagglutinin (FHA) among adults (35–44 years old), young children (4–7 years old), and older children (10–14 years old). Our quantitative analyses revealed that adults had higher seroprevalences of anti-PT IgG and PT-neutralizing antibodies, and similar titers of anti-FHA IgG, compared to the young and older children. Positive correlations were observed between the titers of PT-neutralizing antibodies and anti-PT IgG in all age groups (rs values of 0.326–0.522), although the correlation tended to decrease with age. The ratio of PT-neutralizing antibodies to anti-PT IgG was significantly different when we compared the serum and purified IgG fractions among adults (p = 0.016), although this result was not observed among young and older children. Thus, it appears that some adults had non-IgG immunoglobulins to PT. Our analyses also revealed that adults had high-avidity anti-PT IgG (avidity index: 63.5%, similar results were observed among the children); however, the adults had lower-avidity anti-FHA IgG (37.9%, p < 0.05). It is possible that low-avidity anti-FHA IgG is related to infection with other respiratory pathogens (e.g., Bordetella parapertussis, Haemophilus influenzae, or Mycoplasma pneumoniae), which produces antibodies to FHA-like proteins. Our observations suggest that these adults had been infected with B. pertussis and other pathogen(s) during their adulthood.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

SUMMARY: The purpose of this study was to investigate extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates from pediatric (aged 0 to 3 years) diarrhea patients in Surabaya, Indonesia, where this kind of survey is rare; our study included assessment of their antibiotic susceptibilities, as well as ESBL typing, multilocus sequence typing (MLST), and diarrheagenic E. coli (DEC)-typing. ESBL-producing E. coli were detected in 18.8% of all the samples. Many ESBL-producing E. coli had significantly lower susceptibility to gentamicin (p < 0.0001) and the quinolones nalidixic acid (p=0.004) and ciprofloxacin (p < 0.0001) than non-producers. In ESBL-producing E. coli, 84.0% of strains expressed CTX-M-15 alone or in combination with other ESBL types. MLST revealed that 24.0% of ESBL-producers had sequence type 617, all of which expressed the CTX-M-15 gene; we also detected expression of 3 DEC-related genes: 2 enteroaggregative E. coli genes and 1 enteropathogenic E. coli gene. In conclusion, CTX-M-15-type ESBL-producing E. coli ST617 appear to have spread to Indonesia.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Mycobacterium smegmatis represents one model for studying the biology of its pathogenic relative Mycobacterium tuberculosis. The structural characterization of a M. tuberculosis ortholog protein can serve as a valid tool for the development of molecules active against the M. tuberculosis target. In this context, we report the biochemical and structural characterization of M. smegmatis phosphoribosylpyrophosphate synthetase (PrsA), the ortholog of M. tuberculosis PrsA, the unique enzyme responsible for the synthesis of phosphoribosylpyrophosphate (PRPP). PRPP is a key metabolite involved in several biosynthetic pathways including those for histidine, tryptophan, nucleotides and decaprenylphosphoryl-arabinose, an essential precursor for the mycobacterial cell wall biosynthesis. Since M. tuberculosis PrsA has been validated as a drug target for the development of antitubercular agents, the data presented here will add to the knowledge of the mycobacterial enzyme and could contribute to the development of M. tuberculosis PrsA inhibitors of potential pharmacological interest.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Emerging Infectious Diseases  

Prevalence of pertactin-lacking Bordetella pertussis isolates has been observed worldwide. In Japan, however, we found that the frequency of pertactin-deficient isolates in 2014–2016 (8%) was significantly lower than the frequency in 2005–2007 (41%), 2008–2010 (35%), and 2011–2013 (25%). This reduction was closely associated with changes in genotypes.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Hospital Infection  

Background Nosocomial infections caused by Acinetobacter baumannii international clone II (IC II) can cause severe clinical outcomes. Aim Differential evaluation of bactericidal efficacy of chlorhexidine gluconate (CHX) and benzethonium chloride (BZT) disinfectants against IC II and non-IC II isolates. Methods Minimum inhibitory concentrations (MICs) of CHX and BZT were determined for 137 A. baumannii IC II, 99 non-IC II and 69 non-baumannii isolates, further classified according to MIC values into disinfectant-reduced susceptible (DRS) and disinfectant-susceptible (DS) groups. Time-kill curves and minimum bactericidal concentrations (MBCs) were evaluated for representative isolates in each group. Results CHX and BZT MIC90s for IC II isolates were 100 and 175 mg/L, respectively, but those for non-IC II and non-baumannii isolates were <100 mg/L. Nevertheless, time-kill curves indicated that CHX and BZT reduced live bacterial cell number by 5 log10 for IC II and non-IC II isolates within 30 s when used at 1000 mg/L, comparable to practical use concentrations. CHX MBC at 30 s was 1000 mg/L for IC II and non-IC II isolates, and was not influenced by addition of 3% bovine serum albumin (BSA); BZT MBC at 30 s was 100 mg/L without BSA and increased up to 500 mg/L upon addition of BSA. No significant differences in BSA were found between DRS and DS isolates. Conclusion CHX and BZT were effective against Acinetobacter spp. including IC II at a concentration of 1000 mg/L and exposure for at least 30 s, but their concentrations should be considered carefully to ensure sufficient effects in both clinical and healthcare settings.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：European Journal of Clinical Microbiology and Infectious Diseases  

Acinetobacter baumannii is an important cause of multidrug-resistant hospital acquired infections in the world. Here, we investigate the presence of NDM-1 and other carbapenemases among carbapenem-resistant A. baumannii isolated between August 2010 and December 2014 from three large hospitals in Hanoi, Vietnam. We identified 23/582 isolates (4 %) (11 from hospital A, five from hospital B, and seven from hospital C) that were NDM-1 positive, and among them 18 carried additional carbapenemase genes, including seven isolates carrying NDM-1, IMP-1, and OXA-58 with high MICs for carbapenems. Genotyping indicated that NDM-1 carrying A. baumannii have expanded clonally in these hospitals. Five new STs (ST1135, ST1136, ST1137, ST1138, and ST1139) were identified. One isolate carried NDM-1 on a plasmid belonging to the N-repA replicon type; no NDM-1-positive plasmids were identified in the other isolates. We have shown the extent of the carbapenem resistance and the local clonal spread of A. baumannii carrying NDM-1 in these hospitals; coexistence of NDM-1 and IMP-1 is reported for the first time from Vietnam here, and this will further seriously limit future therapeutic options.

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Future Medicine Ltd  

<jats:p> Aim: To characterize inter- and intra-strain variability of variable-number tandem repeats (VNTRs) in Mycoplasma pneumoniae to determine the optimal multilocus VNTR analysis scheme for improved strain typing. Methods: Whole genome assemblies and next-generation sequencing data from diverse M. pneumoniae isolates were used to characterize VNTRs and their variability, and to compare the strain discriminability of new VNTR and existing markers. Results: We identified 13 VNTRs including five reported previously. These VNTRs displayed different levels of inter- and intra-strain copy number variations. All new markers showed similar or higher discriminability compared with existing VNTR markers and the P1 typing system. Conclusion: Our study provides novel insights into VNTR variations and potential new multilocus VNTR analysis schemes for improved genotyping of M. pneumoniae. </jats:p>

DOI： 10.2217/fmb-2016-0111

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Microbiological Methods  

We evaluated a commercial loop-mediated isothermal amplification (LAMP) assay kit for Bordetella pertussis detection. The LAMP primers were designed to target the ptxP1 allele of the pertussis toxin promoter, but the assay could detect B. pertussis ptxP3 and ptxP8 strains in addition to ptxP1 strains, with high analytical sensitivity.

DOI： 10.1016/j.mimet.2016.12.009

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Microbiological Methods  

The Carba NP test was developed to detect carbapenemase-producing Enterobacteriaceae, and uses imipenem as the reaction substrate. In Japan, IMP-6 metallo-β-lactamase (MBL) producers, which are usually resistant to meropenem but susceptible to imipenem, and IMP-1 MBL producers, which are usually resistant to both carbapenems are prevalent. We performed the Carba NP test with IMP-6 and IMP-1 MBL producers, and both types were detected by the Carba NP test with high sensitivity. All IMP-1 MBL producers were detected by the Carba NP test, but the minimum inhibitory concentrations (MICs) of imipenem varied from 0.25 to > 32 μg/mL, and the time to positivity varied from 0 to 30 min. Time to positivity was significantly correlated with expression levels of blaIMP-1, but not with MICs of imipenem. These results suggested that the Carba NP test can be used as a screening assay for carbapenemase gene expression levels among producers of the same type of carbapenemase. Using this approach, it is possible to determine whether the carbapenem resistance of each carbapenemase-producing Enterobacteriaceae isolate is primarily due to carbapenemase production, or to another mechanism such as outer membrane impermeability.

DOI： 10.1016/j.mimet.2016.12.015

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Pathogens and Disease  

Bordetella pertussis is the etiological agent of pertussis and produces various virulence factors, including pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (PRN), most of which are positively regulated by the BvgAS two-component sensory transduction system. Here, we describe a B. pertussis isolate not expressing PT, FHA and PRN recovered from a pertussis patient. Sequencing revealed that the bvgS gene of this isolate contains a spontaneous mutation (C>A at position 955) causing the proline residue at position 319 of the BvgS protein to be substituted by threonine. Moreover, loss of PT, FHA and PRN expression was completely restored by complementation with a wild-type bvgAS locus, indicating that this non-synonymous substitution in bvgS leads to impaired BvgS function. Our findings indicate that the proline residue at position 319 in this protein plays an essential role in activation of the BvgAS system and, therefore, subsequent expression of Bvg-regulated virulence factors in B. pertussis..

DOI： 10.1093/femspd/ftx011

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
Here, we present the complete genome sequences of
<jats:named-content content-type="genus-species">Mycoplasma pneumoniae</jats:named-content>
KCH-402 and KCH-405, which are
<jats:italic>p1</jats:italic>
gene type 2b and 2c strains, respectively. These strains harbor variations in the
<jats:italic>orf6</jats:italic>
gene, which encodes the cytadherence-related proteins P40 and P90.
</jats:p>

DOI： 10.1128/genomeA.00513-17

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
We report here a draft genome sequence of
<jats:named-content content-type="genus-species">Clostridium botulinum</jats:named-content>
Adk2012 responsible for a foodborne botulism case that occurred in Tottori, Japan, in 2012. Its genome size was 2,904,173 bp, with 46 rRNAs and 54 tRNAs, at a coverage of 14.5×.
</jats:p>

DOI： 10.1128/genomeA.00872-17

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Inflammation and Regeneration  

Background: Bacterial infections cause an increase in the population of hematopoietic stem cells (HSCs). To investigate the downstream factors associated with hematopoietic stem cells, mice are infected with Mycobacterium avium (M. avium). Results: Mycobacterium avium (M. avium) infection induces the enlargement of the spleen and changes in histopathology, including changes to the lineage populations. A dramatic expansion of Lin−c-kit+Sca-1+ (KSL) cells in mouse bone marrow cells and spleen cells was detected 4 weeks after infection with M. avium; however, there was no difference in the engraft activity between infected and un-infected mouse bone marrow cells. We tested the cytokine and cytokine-related gene expression after M. avium infection and found that IFN-γ expression increased and peaked at 4 weeks in both bone marrow and spleen cells. The expression of Sca-1 gene peaked at 4 weeks in the bone marrow but peaked at 2 weeks in spleen cells, although the Sca-1 surface marker peaked at 4 weeks after infection in both bone marrow and spleen cells. Interferon regulatory factor-2 (IRF-2) expression did not change in the bone marrow cells, whereas it decreased in spleen cells at 4 weeks and IRF-1 expression was up-regulated in both bone marrow and spleen cells after infection. However, the up-regulation of IRF-1 was not correlated with IFN-γ expression in the M. avium-infected mouse spleen cells. Conclusions: This finding suggests that the IFN-γ production mediated by M. avium infection alters the population of KSL cells during host defense, and the down-regulation of the IFN-γ response in spleen cells occurs at the late stage after M. avium infection.

DOI： 10.1186/s41232-016-0024-3

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

Carbapenems are a class of last-resort antibiotics; thus, the increase in bacterial carbapenem-resistance is a serious public health threat. Acinetobacter baumannii is one of the microorganisms that can acquire carbapenem-resistance; it causes severe nosocomial infection, and is notoriously difficult to control in hospitals. Recently, a machine-learning approach was first used to analyze the genome sequences of hundreds of susceptible and resistant A. baumannii strains, including those carrying commonly acquired resistant mechanisms, to build a classifier that can predict strain resistance. A complementary approach is to explore novel genetic elements that could be associated with the antimicrobial resistance of strains, independent of known mechanisms. Therefore, we carefully selected A. baumannii strains, spanning various genotypes, from public genome databases, and conducted the first genome-wide association study (GWAS) of carbapenem resistance. We employed a recently developed method, capable of identifying any kind of genetic variation and accounting for bacterial population structure, and evaluated its effectiveness. Our study identified a surface adhesin gene that had been horizontally transferred to an ancestral branch of A. baumannii, as well as a specific region of that gene that appeared to accumulate multiple individual variations across the different branches of carbapenem-resistant A. baumannii strains.

DOI： 10.1038/srep37811

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

Chronic inflammation-associated bone destruction, which is observed in rheumatoid arthritis (RA) and periodontitis, is mediated by excessive osteoclastogenesis. We showed previously that γ-glutamyltranspeptidase (GGT), an enzyme involved in glutathione metabolism, acts as an endogenous activator of such pathological osteoclastogenesis, independent of its enzymatic activity. GGT accumulation is clinically observed in the joints of RA patients, and, in animals, the administration of recombinant GGT to the gingival sulcus as an in vivo periodontitis model induces an increase in the number of osteoclasts. However, the underlying mechanisms of this process remain unclear. Here, we report that Toll-like receptor 4 (TLR4) recognizes GGT to activate inflammation-associated osteoclastogenesis. Unlike lipopolysaccharide, GGT is sensitive to proteinase K treatment and insensitive to polymyxin B treatment. TLR4 deficiency abrogates GGT-induced osteoclastogenesis and activation of NF-κB and MAPK signaling in precursor cells. Additionally, GGT does not induce osteoclastogenesis in cells lacking the signaling adaptor MyD88. The administration of GGT to the gingival sulcus induces increased osteoclastogenesis in wild-type mice, but does not induce it in TLR4-deficient mice. Our findings elucidate a novel mechanism of inflammation-associated osteoclastogenesis, which involves TLR4 recognition of GGT and subsequent activation of MyD88-dependent signaling.

DOI： 10.1038/srep35930

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
We present here the draft whole-genome shotgun sequence of an uncultivated strain SNTW101 of
<jats:named-content content-type="genus-species">Helicobacter suis</jats:named-content>
, which has been maintained in the stomachs of mice. This strain was originally isolated from gastric biopsy specimens of a urea breath test-negative Japanese patient suffering from nodular gastritis.
</jats:p>

DOI： 10.1128/genomeA.00934-16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

<jats:sec sec-type="intro">
<jats:title>Introduction:</jats:title>
<jats:p>
<jats:italic> Mycoplasma hominis</jats:italic> is associated with genito-urinary tract infection and adverse pregnancy outcomes. However, whether the species is a true pathogen or part of the genito-urinary tracts natural flora remains unclear.</jats:p>
</jats:sec>
<jats:sec>
<jats:title>Case presentation:</jats:title>
<jats:p>A 41-year-old pregnant woman was admitted to our hospital at 38 weeks and 5 days of gestation owing to premature rupture of the membranes. The patient delivered by caesarean section. Subsequently, the patient complained of lower abdominal pain and had persistent fever. Enhanced computed tomography revealed pelvic abscesses. Gram staining of pus from the abscess and vaginal secretions indicated presence of polymorphonuclear leucocytes but no pathogens. Cultures on blood agar showed growth of pinpoint-sized colonies in an anaerobic environment within 48 h. Although administration of carbapenem and metronidazole was ineffective and we could not fully drain the abscess, administration of clindamycin led to clinical improvement. The isolates 16S rRNA gene and <jats:italic>yidC</jats:italic> gene sequences exhibited identity with those of <jats:italic>M. hominis.</jats:italic>
</jats:p>
</jats:sec>
<jats:sec sec-type="conclusions">
<jats:title>Conclusion:</jats:title>
<jats:p>Physicians should consider<jats:italic> M. hominis</jats:italic> in cases of pelvic abscesses where Gram staining yields negative results, small colonies are isolated from the abscess and treatment with β-lactam antibiotics is ineffective.</jats:p>
</jats:sec>

DOI： 10.1099/jmmcr.0.005059

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Genome Announcements  

Volume 1, no. 4, e00512-13, 2013. After publication, we detected the presence of a putative cytolethal distending toxin (CDT) gene cluster in Helicobacter fennelliae MRY12-0050. In the last paragraph before the accession numbers, the third sentence should read as follows: "H. cinaedi and H. hepaticus have been reported to express cytolethal distending toxin (CDT), which causes DNA damage to target cells, and a putative CDT gene cluster was also identified in H. fennelliae MRY12-0050 (with the gene product sharing 31%, 36%, and 32% amino acid sequence identity with CdtA, CdtB, and CdtC, respectively, of H. cinaedi strain CCUG 18818)."

DOI： 10.1128/genomeA.00634-16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE  

Bordetella holmesii causes both invasive and respiratory diseases in humans. Although the number of cases of pertussis-like respiratory illnesses due to B. holmesii infection has increased in the last decade worldwide, little is known about the virulence factors of the organism. Here, we analyzed a B. holmesii isolate that forms large aggregates and precipitates in suspension, and subsequently demonstrated that the autoagglutinating isolate is deficient in Bordetella intermediate protein A (BipA) and that this deletion is caused by a frame-shift mutation in the bipA gene. A BipA-deficient mutant generated by homologous recombination also exhibited the autoagglutination phenotype. Moreover, the BipA mutant adhered poorly to an abiotic surface and failed to form biofilms, as did two other B. holmesii autoagglutinating strains, ATCC 51541 and ATCC 700053, which exhibit transcriptional down-regulation of bipA gene expression, indicating that autoagglutination indirectly inhibits biofilm formation. In a mouse intranasal infection model, the BipA mutant showed significantly lower levels of initial lung colonization than did the parental strain (P < 0.01), suggesting that BipA might be a critical virulence factor in B. holmesii respiratory infection. Together, our findings suggest that BipA production plays an essential role in preventing autoagglutination and indirectly promoting biofilm formation by B. holmesii.

DOI： 10.1371/journal.pone.0159999

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley  

An ELISA that measures anti-PT IgG antibody has been used widely for the serodiagnosis of pertussis; however, the IgG-based ELISA is inadequate for patients during the acute phase of the disease because of the slow response of anti-PT IgG antibodies. To solve this problem, we developed a novel IgM-capture ELISA that measures serum anti-Bordetella pertussis Vag8 IgM levels for the accurate and early diagnosis of pertussis. First, we confirmed that Vag8 was highly expressed in all B. pertussis isolates tested (n=30), but little or none in other Bordetella species, and that DTaP vaccines did not induce anti-Vag8 IgG antibodies in mice (i.e. the antibody level could be unaffected by the vaccination). To determine the immune response to Vag8 in B. pertussis infection, anti-Vag8 IgM levels were compared between 38 patients (acute phase of pertussis) and 29 healthy individuals using the anti-Vag8 IgM-capture ELISA. The results revealed that the anti-Vag8 IgM levels were significantly higher in the patients compared with the healthy individuals (P<0.001). ROC analysis also showed that the anti-Vag8 IgM-capture ELISA has higher diagnostic accuracy (AUC, 0.92) than a commercial anti-PT IgG ELISA kit. Moreover, it was shown that anti-Vag8 IgM antibodies were induced earlier than anti-PT IgG antibodies on sequential patients' sera. These data indicate that our novel anti-Vag8 IgM-capture ELISA is a potentially useful tool for making the accurate and early diagnosis of B. pertussis infection.

DOI： 10.1111/1348-0421.12378

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

In 2011, a multidrug-resistant <i>Acinetobacter baumannii</i> (MDRAB) outbreak occurred at a Japanese critical care center (CCC) in a tertiary care hospital. Multidrug-resistance in Japan is defined as resistance to the antimicrobials amikacin, carbapenem, and fluoroquinolone. We conducted a retrospective epidemiological investigation of this outbreak to identify the risk factors for MDRAB respiratory tract acquisition in this hospital. Cases were defined as hospitalized patients with MDRAB-positive cultures at least 3 days post admission to the CCC between June 1, 2011 and April 20, 2012. Fifteen MDRAB cases were identified, including 3 with infection and 12 with colonization. This case-control study demonstrated that hypoalbuminemia along with carbapenem administration were associated with MDRAB respiratory tract acquisition. Pulsed-field gel electrophoresis analysis and multilocus sequence typing using MDRAB isolates suggested a clonal dissemination of MDRAB strains with sequence type 74 occurred primarily among patients admitted to the CCC. From April 16, 2012, a decreased isolation rate of MDRAB in the hospital occurred after the implementation of the following infection control measures: closing the emergency room, discontinuing admission to the CCC, isolating patients with MDRAB colonization or infection to single room status, and conducting environmental cleaning. No MDRAB cases were detected between March 23 and April 20, 2012.

DOI： 10.7883/yoken.JJID.2015.049

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/femspd/ftv109

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

We report an 8-year-old patient with catheter-related bacteremia caused by linezolid-resistant Staphylococcus epidermidis that was isolated after the long-term, repeated use of linezolid. Three S. epidermidis strains isolated from this patient were bacteriologically analyzed. While the strain isolated prior to linezolid initiation was susceptible to linezolid, two strains after linezolid therapy displayed low-level linezolid susceptibility (MIC, 4 mg/L) and linezolid resistance (MIC, 16 mg/L). T2500A mutation in two copies and G2575T mutations in three copies of 23S rRNA were detected in the low-susceptible strain and the resistant strain, respectively. Linezolid-resistant S. epidermidis infection is rare, but may occur with the long-term administration of linezolid.

DOI： 10.1016/j.jiac.2015.10.004

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担当区分：筆頭著者,　最終著者,　責任著者   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

Volume 69, no. 2, p. 143&ndash;148, 2016. Page 143, Title should appear as shown below.<br><br>An Epidemiological Investigation of a Nosocomial Outbreak of Multidrug-Resistant <i>Acinetobacter baumannii</i> in a Critical Care Center in Japan, 2011&ndash;2012

DOI： 10.7883/yoken.JJID.2016.E002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
<jats:named-content content-type="genus-species">Pseudomonas aeruginosa</jats:named-content>
causes serious intractable infections in humans and animals. Bacteriophage (phage) therapy has been applied to treat
<jats:named-content content-type="genus-species">P. aeruginosa</jats:named-content>
infections, and phages belonging to the PB1-like virus genus in the
<jats:named-content content-type="genus-species">Myoviridae</jats:named-content>
family have been used as therapeutic phages. To achieve safer and more effective phage therapy, the use of preadapted phages is proposed. To understand in detail such phage preadaptation, the short-term antagonistic evolution of bacteria and phages should be studied. In this study, the short-term antagonistic evolution of bacteria and PB1-like phage was examined by studying phage-resistant clones of
<jats:named-content content-type="genus-species">P. aeruginosa</jats:named-content>
strain PAO1 and mutant PB1-like phages that had recovered their infectivity. First, phage KPP22 was isolated and characterized; it was classified as belonging to the PB1-like virus genus in the
<jats:named-content content-type="genus-species">Myoviridae</jats:named-content>
family. Subsequently, three KPP22-resistant PAO1 clones and three KPP22 mutant phages capable of infecting these clones were isolated in three sets of
<jats:italic>in vitro</jats:italic>
experiments. It was shown that the bacterial resistance to phage KPP22 was caused by significant decreases in phage adsorption and that the improved infectivity of KPP22 mutant phages was caused by significant increases in phage adsorption. The KPP22-resistant PAO1 clones and the KPP22 mutant phages were then analyzed genetically. All three KPP22-resistant PAO1 clones, which were deficient for the O5 antigen, had a common nonsense mutation in the
<jats:italic>wzy</jats:italic>
gene. All the KPP22 mutant phage genomes showed the same four missense mutations in the open reading frames
<jats:italic>orf060</jats:italic>
,
<jats:italic>orf065</jats:italic>
, and
<jats:italic>orf086</jats:italic>
. The information obtained in this study should be useful for further development of safe and efficient phage therapy.
</jats:p>
<jats:p>
<jats:bold>IMPORTANCE</jats:bold>
<jats:named-content content-type="genus-species">Pseudomonas aeruginosa</jats:named-content>
causes serious intractable infections in humans and animals; bacteriophage (phage) therapy has been utilized to treat
<jats:named-content content-type="genus-species">P. aeruginosa</jats:named-content>
infections, and phages that belong to the PB1-like virus genus in the family
<jats:named-content content-type="genus-species">Myoviridae</jats:named-content>
have been used as therapeutic phages. The preadapted phage is trained in advance through the antagonistic evolution of bacteria and phage and is proposed to be used to achieve safer and more effective phage therapy. In this study, to understand the phage preadaptation, the
<jats:italic>in vitro</jats:italic>
short-term antagonistic evolution was studied using
<jats:named-content content-type="genus-species">P. aeruginosa</jats:named-content>
strain PAO1 and the newly isolated PB1-like phage KPP22. Phage KPP22 was characterized, and the molecular framework regarding the phage preadaptation of KPP22 was elucidated. The importance of study of antagonistic evolution of bacteria and phage in phage therapy is discussed.
</jats:p>

DOI： 10.1128/AEM.00090-16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
The emergence of multiple-carbapenemase-producing
<jats:italic>Acinetobacter</jats:italic>
strains has been a serious concern during the past decade. Here, we report the draft genome sequence of an
<jats:named-content content-type="genus-species">Acinetobacter baumannii</jats:named-content>
strain isolated from a Japanese patient with three acquired carbapenemase genes:
<jats:italic>bla</jats:italic>
<jats:sub>NDM-1</jats:sub>
,
<jats:italic>bla</jats:italic>
<jats:sub>TMB-1</jats:sub>
, and
<jats:italic>bla</jats:italic>
<jats:sub>OXA-58</jats:sub>
.
</jats:p>

DOI： 10.1128/genomeA.01290-16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Genome Announcements  

Some bacterial species of the genus Tenacibaculum, including Tenacibaculum ovolyticum, have been known as fish pathogens in the sea. So far, the only published genome sequence for this genus is for Tenacibaculum dicentrarchi, which could also be a fish pathogen. Strain da5A-8, showing 100% identity to the 16S rRNA gene sequence of T. ovolyticum DSM 18103T, was isolated from seawater at a depth of 344min Kochi, Japan, and grew optimally at 10 to 20°C. The genome sequence of strain da5A-8 revealed the possible virulence genes commonly observed in the genus Tenacibaculum.

DOI： 10.1128/genomeA.00644-16

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：川崎医科大学  

Although fluoroquinolone (FQ) has been used for the treatment of various bacterial infectious diseases, its continued use has been problematic given the appearance of FQ-resistant bacteria. However, the recent discovery of four plasmid-mediated quinolone resistance (PMQR) genes comprising qnr, aac(6')Ib-cr, qepA and OqxAB since 1998 has provided insights in the area of FQ-resistance. For practical detection of qepA in microbiology laboratory, a specific, simple, rapid and cost-effective isothermal amplification method designated as LAMP is the good candidate to use. In this study, the development of a new detection method using LAMP to identify qepA1, one variant of the qepA gene, was tried. As the results, the LAMP method using a qepA1-specific LAMP primer set comprising five primerscould detect all four qepA1-positive strains in addition to 17 qepA1-negative strains. The LAMP method is clearly much more advantageous for use in clinical laboratories. Furthermore, the time and accuracy benefits allow for the selection of antibiotics in a clinical setting.

DOI： 10.11482/KMJ-E42(1)25

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その他リンク： http://id.nii.ac.jp/1162/00000578/

記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：New Microbes and New Infections  

Between January 2013 and December 2014, we conducted laboratory-based surveillance of pertussis using multitarget real-time PCR, which discriminates among Bordetella pertussis, Bordetella parapertussis, Bordetella holmesii and Mycoplasma pneumoniae. Of 355 patients clinically diagnosed with pertussis in Japan, B. pertussis, B. parapertussis and M. pneumoniae were detected in 26% (n = 94), 1.1% (n = 4) and 0.6% (n = 2), respectively, whereas B. holmesii was not detected. It was confirmed that B. parapertussis and M. pneumoniae are also responsible for causing pertussis-like illness. The positive rates for B. pertussis ranged from 16% to 49%, depending on age. Infants aged ≤ 3 months had the highest rate (49%), and children aged 1 to 4 years had the lowest rate (16%, p < 0.01 vs. infants aged ≤ 3 months). Persons aged 10 to 14 and 15 to 19 years also showed high positive rates (29% each); the positive rates were not statistically significant compared with that of infants aged ≤ 3 months (p ≥ 0.06). Our observations indicate that similar to infants, preteens and teens are at high risk of B. pertussis infection.

DOI： 10.1016/j.nmni.2015.10.001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

Global spread and evolutionary links of an epidemic Clostridium difficile strain (PCR-ribotype 027) have been noted in recent decades. However, in Japan, no outbreaks caused by type 027 have been reported to date. A total of 120 C. difficile isolates from patients at 15 hospitals during non-outbreak seasons between 2011 and 2013 as well as 18 and 21 isolates collected from two hospitals in 2010 and 2009, respectively, in outbreak periods in Japan, were examined. Among these 120 isolates, Japan-ribotypes smz and ysmz (subtype variant of smz) were the most predominant (39.2%) followed by Japan-ribotype trf (15.8%). Types smz/ysmz and trf were also concurrently predominant at two hospitals in the outbreak settings. Out of the five binary toxin-positive isolates observed, only one was PCR-ribotype 027 and another PCRribotype 078. Type smz was later found to correspond to PCR-ribotype 018. High rates of resistance against atifloxacin, moxifloxacin, erythromycin and clindamycin were observed in the PCR-ribotype 018 isolates. Interestingly, all trf isolates were toxin A-negative, toxin B-positive, but they did not correspond to PCR-ribotype 017, thus being assigned a new ribotype (PCRribotype 369). In conclusion, PCR-ribotypes 018 (smz) and 369 (trf) were identified as major circulating strains in both outbreak and non-outbreak settings in Japan. Given their epidemiological relevance, molecular investigations are warranted to clarify potential evolutionary links with related strains found elsewhere, such as PCR-ribotypes 018 and 017 from Europe and North America.

DOI： 10.1099/jmm.0.000149

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Diagnostic Microbiology and Infectious Disease  

We developed a novel chromogenic method, Penta-well test, which enables the rapid detection and classification of β-lactamases in clinical Enterobacteriaceae isolates. This test is based on a combination of nitrocefin and 3 β-lactamase inhibitors specific to classes A, B, and/or C, with nitrocefin hydrolysis by β-lactamases being assessed by optical density measurements at 490. nm. When the cutoff value for each β-lactamase class was determined (0.09, 0.4, and 0.55 for class A, class B, and class C β-lactamase producers, respectively), the sensitivity and specificity of classification were 93.5% and 68.8% for class A, 93.8% and 100% for class B, and 86.7% and 100% for class C, respectively. Moreover, this method allowed accurate β-lactamase classification in 20 of 23 (87.0%) isolates producing plural class of β-lactamases. Thus, the Penta-well test can provide information that would be useful in the accurate detection and classification of β-lactamases produced by causative bacteria.

DOI： 10.1016/j.diagmicrobio.2015.06.002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Scientific Reports  

Until the early 1990s, incidences of Mycoplasma pneumoniae (MP) infection showed three to five year epidemic cycles in multiple countries, however, the mechanism for the MP epidemic cycle has not been understood. Here, we investigate the determinant of periodicity in MP incidence by employing a mathematical model describing MP transmission dynamics. Three candidates for the determinant of periodicity were evaluated: school-Term forcing, minor variance in the duration of immunity, and epidemiological interference between MP serotypes. We find that minor variation in the duration of immunity at the population level must be considered essential for the MP epidemic cycle because the MP cyclic incidence pattern did not replicate without it. Minor variation, in this case, is a less dispersed distribution for the duration of immunity than an exponential distribution. Various lengths of epidemic cycles, including cycles typically found in nature, e.g. three to five year cycles, were also observed when there was minor variance in the duration of immunity. The cyclic incidence pattern is robust even if there is epidemiological interference due to cross-immune protection, which is observed in the epidemiological data as negative correlation between epidemics per MP serotype.

DOI： 10.1038/srep14473

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley  

In the past year, a substantial number of (putative) novel Helicobacter species have been described, including Helicobacter himalayensis colonizing the Himalayan marmot and Helicobacter apodemus, colonizing the Korean striped field mouse. In addition, a putative novel gastric Helicobacter species was identified in wild gorillas and chimpanzees, for which the name "Candidatus H. homininae" was proposed. A high incidence of gastric non-H. pylori Helicobacter infection was described in China and multiple case reports have described the involvement of enterohepatic Helicobacter species, especially Helicobacter cinaedi, in a wide range of diseases. Several studies in rodent models further elucidated the mechanisms underlying the development of gastric mucosa-associated lymphoid tissue lymphoma during infection with gastric non-H. pylori Helicobacters. The effects of infection with gastric Helicobacters on the development of neuroinflammation were investigated and several enterohepatic Helicobacter species were shown to affect the composition of the gut microbiota, to influence vaccine efficiency as well as the progression of cancer in distant sites of the body.

DOI： 10.1111/hel.12259

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Protein Expression and Purification  

We recently demonstrated that the Rv2613c protein from Mycobacterium tuberculosis H37Rv is a novel diadenosine 5′,5⌄-P1,P4-tetraphosphate (Ap4A) phosphorylase (MtAPA) that forms a tetramer. Mycobacterium avium and Mycobacterium smegmatis express proteins named MAV-3489 and MSMEG-2932, respectively, that are homologous to MtAPA. Here we showed that the MAV-3489 and MSMEG-2932 proteins possess Ap4A phosphorylase activity and enzymatic properties similar to those of MtAPA. Furthermore, gel-filtration column chromatography revealed that MAV-3489 and MSMEG-2932 assembled into homotetramers in solution, indicating that they may also form unique Ap4A-binding sites composed of tetramers.

DOI： 10.1016/j.pep.2015.04.010

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：国立感染症研究所 Japanese Journal of Infectious Diseases 編集委員会  

Since Group B Streptococcus (GBS, Streptococcus agalactiae) clinical isolates are believed to be uniformly susceptible to b-lactams, penicillin G has been used as the first-line agent for the prevention and treatment of GBS infections. However, the existence and characteristics of GBS isolates with reduced penicillin susceptibility (PRGBS) have recently been reported in Japan. Moreover, the sequence type (ST) 458 is predominant among the PRGBS in Japan. Although the majority of the PRGBS isolates in Japan have been recovered from respiratory specimens of adults, no information on the genotype of these isolates is available. Therefore, whether ST458 predominates among GBS isolates obtained from such specimens is not known. In this study, we characterized the STs of 38 penicillin-susceptible GBS isolates (PSGBS) recovered from respiratory specimens and compared them to the reported PRGBS STs. ST458, the predominant ST among the PRGBS isolates studied (10/19, 53z), was not found in the PSGBS isolates. Thirty-six PSGBS isolates belonged to the ST1/19/10 group (includes 6 different STs), and the remaining 2 isolates belonged to that of ST23. Further, the PRGBS isolates were divided into the ST1 (3 STs), and ST23 (2 STs) groups. ST458 was not predominant among the PSGBS isolates recovered from respiratory specimens in Japan and may therefore be specific to the PRGBS. Thus, the ST distribution of the PRGBS isolates does not reflect that of the PSGBS.

DOI： 10.7883/yoken.JJID.2014.387

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：一般社団法人 日本感染症学会  

近年，小児科を中心にマクロライド耐性<i>Mycoplasma pneumoniae </i>株の増加が報告されており，内科領域においても増加がみられている．2008～2012 年に肺炎マイコプラズマが検出された70 例の市中肺炎患者を対象に，抗菌薬感受性および臨床経過に関して検討を行った．2003～2006 年の検出株ではマクロライド耐性株を認めなかったが，今回の検討では，<i>M. pneumoniae</i>70 株中46 株（65.7％）がマクロライド系薬剤耐性23 SrRNA 遺伝子変異株（A2063G）であり，近年の耐性化傾向を裏付けるものであった． マクロライド耐性株の臨床経過について検討を行ったが，マクロライド耐性株検出例は感受性株検出例に比較し，発熱期間の延長を認めたものの入院期間の有意差は認めなかった．初回使用抗菌薬に感受性のあった場合の発熱期間とマクロライド感受性株の発熱期間はともに約7 日であり，発熱期間は，マクロライド耐性か否かより，初回抗菌薬に感受性があるかどうかに依存すると考えられた．また，耐性株であっても，感受性のある抗菌薬未使用で軽快した例も多く認められており，重篤化や入院期間の延長は認めなかった．よって，マクロライド系薬は，現在においてもマイコプラズマ肺炎の第1 選択薬として可と考えられ，発熱が続き全身状態が改善しない時に薬剤変更を検討してもよいと思われる．

DOI： 10.11150/kansenshogakuzasshi.89.458

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：European Journal of Clinical Microbiology and Infectious Diseases  

This study sought to monitor the presence of carbapenem-resistant Enterobacteriaceae (CRE) and the proportion New Delhi metallo-beta-lactamase 1 (NDM-1)-producing bacteria between August 2010 and December 2012 in a surgical hospital in Vietnam. We identified 47 CRE strains from a total of 4,096 Enterobacteriaceae isolates (1.1 %) that were NDM-1-positive from 45 patients admitted to 11 different departments, with the majority being from the urology department. The NDM-1 gene was found in seven different species. Genotyping revealed limited clonality of NDM-1-positive isolates. Most of the isolates carried the NDM-1 gene on a plasmid and 17.8 % (8/45) of those were readily transferable. We found five patients at admission and one patient at discharge with NDM-1-positive bacteria in their stool. From 200 screening environmental hospital samples, five were confirmed to be NDM-1-positive and included Acinetobacter species (n = 3) and Enterobacter aerogenes (n = 2). The results reveal that NDM-1-producing Enterobacteriaceae are commonly isolated in patients admitted to a Vietnamese surgical hospital and are also detected in the hospital environment.

DOI： 10.1007/s10096-015-2345-6

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Infectious Diseases  

Objectives: The present study was designed to determine the genotypes of circulating Bordetella pertussis in the Philippines by direct molecular typing of clinical specimens. Methods: Nasopharyngeal swabs (NPSs) were collected from 50 children hospitalized with pertussis in three hospitals during 2012-2014. Multilocus variable-number tandem repeat analysis (MLVA) was performed on the DNA extracts from NPSs. B. pertussis virulence-associated allelic genes (. ptxA, prn, and fim3) and the pertussis toxin promoter, ptxP, were also investigated by DNA sequence-based typing. Results: Twenty-six DNA extracts yielded a complete MLVA profile, which were sorted into 10 MLVA types. MLVA type 34 (MT34), which is rare in Australia, Europe, Japan, and the USA, was the predominant strain (50%). Seven MTs (MT29, MT32, MT33, and MT283-286, total 42%) were single-locus variants of MT34, while two (MT141 and MT287, total 8%) were double-locus variants of MT34. All MTs had the combination of virulence-associated allelic genes, ptxP1-. ptxA1-. prn1-. fim3A. Conclusions: The B. pertussis population in the Philippines comprises genetically related strains. These strains are markedly different from those found in patients from other countries where acellular pertussis vaccines are used. The differences in vaccine types between these other countries and the Philippines, where the whole-cell vaccine is still used, may select for distinct populations of B. pertussis.

DOI： 10.1016/j.ijid.2015.04.001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Genome Announcements  

Ralstonia mannitolilytica, a Gram-negative aerobic bacterium, is an opportunistic human pathogen that is becoming more common in cases of nosocomial infections. We report for the first time the whole-genome sequence analysis of R. mannitolilytica strain MRY14-0246, which carries the intrinsic OXA-443/OXA-22-like and OXA-444/OXA-60-like β-lactamase genes and is resistant to meropenem.

DOI： 10.1128/genomeA.00405-15

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbial Pathogenesis  

Diarrhea and pseudomembrane colitis caused by Clostridium difficile infection is a global health concern because of the high recurrence rate after standard antibiotic therapy. Vaccination presents a powerful countermeasure against disease recurrence. In this study, mice vaccinated with the nontoxigenic C.difficile membrane fraction generated a marked immune response to the antigen, as demonstrated by the serum IgG and intestinal fluid IgA levels. Significantly, pretreatment with harvested IgG- and IgA-containing fluids was sufficient to prevent invitro adhesion of C.difficile to human Caco-2 intestinal cells. These results highlight the potential of nontoxigenic C.difficile membrane fraction as a vaccine candidate for C.difficile infection.

DOI： 10.1016/j.micpath.2015.03.001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Antimicrobial Chemotherapy  

Objectives: Linezolid has been reported to remain active against 98% of staphylococci with resistance identified in 0.05% of Staphylococcus aureus and 1.4% of CoNS. The objective of this study was to characterize the linezolidresistance mechanisms in the linezolid-resistant CoNS strains isolated in Japan. Methods: Staphylococcus capitis strains exhibiting linezolid MICs > 8 mg/L isolated from inpatients between 2012 and 2014 were screened for cfr and mutations in 23S rRNA, L3 and L4 by PCR/sequencing. Isolates were also examined for mutations in the rlmN gene. Results: S. capitis had six 23S rRNA alleles. Five S. capitis isolates displayed linezolid MICs of 8, 16 and 32 mg/L. G2576U mutations were detected in three, four or five copies of 23S rRNA in all isolates. In two isolates exhibiting the highest linezolid MIC (32 mg/L) there was a large deletion in a single copy of 23S rRNA. Repeated 10 bp sequences were found in both 16S and 23S rRNAs, suggesting deletion by recombination between the repeats. One isolate had the mutation Ala-142→Thr in the ribosomal protein L3. All linezolid-resistant isolates also demonstrated mutations in the gene encoding RlmN methyltransferase, leading to Thr-62→Met and Gly-148→Ser. Conclusions: Multiple mechanisms appeared to be responsible for the elevated linezolid resistance in S. capitis isolates: a G2576U mutation in different numbers of copies of 23S rRNA, loss of a single copy of 23S rRNA and a mutation in the ribosomal protein L3, suggesting the accumulation of independent mutational events.

DOI： 10.1093/jac/dku443

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担当区分：筆頭著者,　最終著者,　責任著者   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：公益社団法人 日本薬学会  

IMP-2, a subclass B1 metallo-β-lactamase (MBL), is a Zn(II)-containing hydrolase. This hydrolase, involved in antibiotic resistance, catalyzes the hydrolysis of the C-N bond of the β-lactam ring in β-lactam antibiotics such as benzylpenicillin and imipenem. The crystal structure of IMP-2 MBL from Acinetobacter spp. was determined at 2.3Å resolution. This structure is analogous to that of subclass B1 MBLs such as IMP-1 and VIM-2. Comparison of the structures of IMP-1 and IMP-2, which have an 85% amino acid identity, suggests that the amino acid substitution at position 68 on a β-strand (β3) (Pro in IMP-1 versus Ser in IMP-2) may be a staple factor affecting the flexibility of loop 1 (comprising residues at positions 60-66; EVNGWGV). In the IMP-1 structure, loop 1 adopts an open, disordered conformation. On the other hand, loop 1 of IMP-2 forms a closed conformation in which the side chain of Trp64, involved in substrate binding, is oriented so as to cover the active site, even though there is an acetate ion in the active site of both IMP-1 and IMP-2. Loop 1 of IMP-2 has a more flexible structure in comparison to IMP-1 due to having a Ser residue instead of the Pro residue at position 68, indicating that this difference in sequence may be a trigger to induce a more flexible conformation in loop 1.

DOI： 10.1248/bpb.b14-00594

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：一般社団法人 日本内科学会  

A 56-year-old previously healthy man was hospitalized due to a 10-day history of neck pain and an elevated C-reactive protein level. Gram-negative spiral bacilli were isolated from his blood, and Helicobacter cinaedi was confirmed using 16S rRNA sequencing. The infectious focus was not identified by initial cervical magnetic resonance imaging (MRI); however, repeated MRI demonstrated prominent high signal intensity in the entire region of the C6-C7 vertebrae and C6/C7 disc space. Furthermore, fluorodeoxyglucose-positron emission tomography/computed tomography showed no significant uptake, other than in the C6-C7 region. The patient was successfully treated with ceftriaxone for six weeks without sequelae.

DOI： 10.2169/internalmedicine.54.4574

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：一般社団法人 日本内科学会  

A 43-year-old man was referred to our hospital for an acute-onset fever and left flank pain. He had been previously diagnosed with lymphangioma, and abdominal computed tomography showed pararenal cysts with fat stranding around the left kidney, of which infection was subsequently confirmed on magnetic resonance imaging. Gram-negative spiral bacilli were isolated from two sets of blood cultures, and Helicobacter cinaedi was identified using 16S rRNA sequencing. The patient was successfully treated with ceftriaxone therapy without recurrence. A multilocus sequence typing analysis indicated the current H. cinaedi strain differed from previous strains isolated in Japan.

DOI： 10.2169/internalmedicine.54.3991

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Oxford University Press (OUP)  

<jats:title>Abstract</jats:title>
<jats:p>Diadenosine 5′,5′′′-P1,P4-tetraphosphate (Ap4A) phosphorylase from Mycobacterium tuberculosis H37Rv (MtAPA) belongs to the histidine triad motif (HIT) superfamily, but is the only member with an alanine residue at position 149 (Ala-149). Enzymatic analysis revealed that the Ala-149 deletion mutant displayed substrate specificity for diadenosine 5′,5′′′-P1,P5-pentaphosphate and was inactive on Ap4A and other substrates that are utilized by the wild-type enzyme.</jats:p>

DOI： 10.1080/09168451.2014.973364

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1186/s40560-015-0081-8

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

DOI： 10.1128/AAC.04265-14

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

<jats:p>A series of clinical isolates of drug-resistant (DR)<jats:italic>Acinetobacter baumannii</jats:italic>with diverse drug susceptibility was detected from eight patients in the emergency intensive care unit of Tokai University Hospital. The initial isolate was obtained in March 2010 (<jats:italic>A. baumannii</jats:italic>Tokai strain 1); subsequently, seven isolates were obtained from patients (<jats:italic>A. baumannii</jats:italic>Tokai strains 2–8) and one isolate was obtained from an air-fluidized bed used by five of the patients during the 3 months from August to November 2011. The isolates were classified into three types of antimicrobial drug resistance patterns (RRR, SRR and SSR) according to their susceptibility (S) or resistance (R) to imipenem, amikacin and ciprofloxacin, respectively. Genotyping of these isolates by multilocus sequence typing revealed one sequence type, ST208, whilst that by a DiversiLab analysis revealed two subtypes. All the isolates were positive for<jats:italic>bla</jats:italic><jats:sub>OXA-51-like</jats:sub>and<jats:italic>bla</jats:italic><jats:sub>OXA-66</jats:sub>, as assessed by PCR and DNA sequencing.<jats:italic>A. baumannii</jats:italic>Tokai strains 1–8 and 10 (RRR, SRR and SSR) had quinolone resistance-associated mutations in<jats:italic>gyrA/parC</jats:italic>, as revealed by DNA sequencing. The IS<jats:italic>Aba1</jats:italic>upstream of<jats:italic>bla</jats:italic><jats:sub>OXA-51-like</jats:sub>and aminoglycoside resistance-associated gene,<jats:italic>armA</jats:italic>, were detected in<jats:italic>A. baumannii</jats:italic>Tokai strains 1–7 and 10 (RRR and SRR) as assessed by PCR. Among the genes encoding resistance–nodulation–division family pumps (<jats:italic>adeB, adeG</jats:italic>and<jats:italic>adeJ</jats:italic>) and outer-membrane porins (<jats:italic>oprD</jats:italic>and<jats:italic>carO</jats:italic>), overexpression of<jats:italic>adeB</jats:italic>and<jats:italic>adeJ</jats:italic>and suppression of<jats:italic>oprD</jats:italic>and<jats:italic>carO</jats:italic>were seen in isolates of<jats:italic>A. baumannii</jats:italic>Tokai strain 2 (RRR), as assessed by real-time PCR. Thus, the molecular characterization of a series of isolates of DR<jats:italic>A. baumannii</jats:italic>revealed the outbreak of ST208 and diverse antimicrobial drug susceptibilities, which almost correlated with differential gene alterations responsible for each type of drug resistance.</jats:p>

DOI： 10.1099/jmm.0.077503-0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
Genetic characterization was performed for 10 group I
<jats:named-content content-type="genus-species">Clostridium botulinum</jats:named-content>
strains isolated from botulism cases in Japan between 2006 and 2011. Of these, 1 was type A, 2 were type B, and 7 were type A(B) {carrying a silent
<jats:italic>bont</jats:italic>
/
<jats:italic>B</jats:italic>
[
<jats:italic>bont</jats:italic>
/
<jats:italic>(B)</jats:italic>
] gene} serotype strains, based on botulinum neurotoxin (BoNT) production. The type A strain harbored the subtype A1 BoNT gene (
<jats:italic>bont</jats:italic>
/
<jats:italic>A1</jats:italic>
), which is associated with the
<jats:italic>ha</jats:italic>
gene cluster. The type B strains carried
<jats:italic>bont</jats:italic>
/
<jats:italic>B5</jats:italic>
or
<jats:italic>bont</jats:italic>
/
<jats:italic>B6</jats:italic>
subtype genes. The type A(B) strains carried
<jats:italic>bont</jats:italic>
/
<jats:italic>A1</jats:italic>
identical to that of type A(B) strain NCTC2916. However,
<jats:italic>bont</jats:italic>
/
<jats:italic>(B)</jats:italic>
genes in these strains showed single-nucleotide polymorphisms (SNPs) among strains. SNPs at 2 nucleotide positions of
<jats:italic>bont</jats:italic>
/
<jats:italic>(B)</jats:italic>
enabled classification of the type A(B) strains into 3 groups. Pulsed-field gel electrophoresis (PFGE) and multiple-locus variable-number tandem-repeat analysis (MLVA) also provided consistent separation results. In addition, the type A(B) strains were separated into 2 lineages based on their plasmid profiles. One lineage carried a small plasmid (5.9 kb), and another harbored 21-kb plasmids. To obtain more detailed genetic information about the 10 strains, we sequenced their genomes and compared them with 13 group I
<jats:named-content content-type="genus-species">C. botulinum</jats:named-content>
genomes in a database using whole-genome SNP analysis. This analysis provided high-resolution strain discrimination and enabled us to generate a refined phylogenetic tree that provides effective traceability of botulism cases, as well as bioterrorism materials. In the phylogenetic tree, the subtype B6 strains, Okayama2011 and Osaka05, were distantly separated from the other strains, indicating genomic divergence of subtype B6 strains among group I strains.
</jats:p>

DOI： 10.1128/AEM.02134-14

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Wiley  

The laboratory diagnostic methods for Clostridium difficile infection (CDI) include toxigenic culture, enzyme immunoassays (EIAs) to detect the toxins of C. difficile, and nucleic acid amplification tests (NAATs) to detect C. difficile toxin genes, but each of these methods has disadvantages; toxigenic cultures require a long time to produce results, EIAs have low sensitivity, and NAATs that target DNA cannot distinguish vegetative cells from spores and dead cells. Here we report a new detection method that uses reverse transcription polymerase chain reaction to target the toxin-gene transcripts. This method was able to specifically detect the vegetative cells of toxigenic C. difficile in fecal samples in spike tests, with a minimum detection limit of 5×102 colony-forming units per 100mg of stool specimen. The performance of this method was also demonstrated in a pilot scale evaluation using clinical fecal specimens, which showed that this method may be more sensitive than EIA and requires a shorter time than toxigenic culture. This method could potentially be applied in the clinical laboratory to detect C. difficile in fecal specimens. The ability of this method to discriminate the presence of vegetative cells from spores and dead cells could help to further the understanding of CDI.

DOI： 10.1111/1348-0421.12189

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

Using the loop-mediated isothermal amplification (LAMP) method, we developed a rapid assay for detection of 16S rRNA methylase genes (rmtA, rmtB, and armA), and investigated 16S rRNA methylase-producing strains among clinical isolates. Primer Explorer V3 software was used to design the LAMP primers. LAMP primers were prepared for each gene, including two outer primers (F3 and B3), two inner primers (FIP and BIP), and two loop primers (LF and LB). Detection was performed with the Loopamp DNA amplification kit. For all three genes (rmtA, rmtB, and armA), 102 copies/tube could be detected with a reaction time of 60 min. When nine bacterial species (65 strains saved in National Institute of Infectious Diseases) were tested, which had been confirmed to possess rmtA, rmtB, or armA by PCR and DNA sequencing, the genes were detected correctly in these bacteria with no false negative or false positive results. Among 8447 clinical isolates isolated at 36 medical institutions, the LAMP method was conducted for 191 strains that were resistant to aminoglycosides based on the results of antimicrobial susceptibility tests. Eight strains were found to produce 16S rRNA methylase (0.09%), with rmtB being identified in three strains (0.06%) of 4929 isolates of Enterobacteriaceae, rmtA in three strains (0.10%) of 3284 isolates of Pseudomonas aeruginosa, and armA in two strains (0.85%) of 234 isolates of Acinetobacter spp. At present, the incidence of strains possessing 16S rRNA methylase genes is very low in Japan. However, when Gram-negative bacteria showing high resistance to aminoglycosides are isolated by clinical laboratories, it seems very important to investigate the status of 16S rRNA methylase gene-harboring bacilli and monitor their trends among Japanese clinical settings.

DOI： 10.1016/j.jiac.2014.08.013

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担当区分：筆頭著者,　最終著者,　責任著者   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
The number of reports concerning
<jats:named-content content-type="genus-species">Escherichia coli</jats:named-content>
clinical isolates that produce glutathione
<jats:italic>S</jats:italic>
-transferases responsible for fosfomycin resistance (FR-GSTs) has been increasing. We have developed a disk-based potentiation test in which FR-GST producers expand the growth inhibition zone around a Kirby-Bauer disk containing fosfomycin in combination with sodium phosphonoformate (PPF). PPF, an analog of fosfomycin, is a transition-state inhibitor of FosA
<jats:sup>PA</jats:sup>
, a type of FR-GST from
<jats:named-content content-type="genus-species">Pseudomonas aeruginosa</jats:named-content>
. Considering its mechanism of action, PPF was expected to inhibit a variety of FR-GSTs. In the presence of PPF, zone enlargement around the disk containing fosfomycin was observed for FosA3-, FosA4-, and FosC2-producing
<jats:named-content content-type="genus-species">E. coli</jats:named-content>
clinical isolates. Moreover, the growth inhibition zone was remarkably enlarged when the Mueller-Hinton (MH) agar plate contained 25 μg/ml glucose-6-phosphate (G6P). When we retrospectively tested 12 fosfomycin-resistant (MIC, ≥256 μg/ml)
<jats:named-content content-type="genus-species">E. coli</jats:named-content>
clinical isolates from our hospital with the potentiation test, 6 FR-GST producers were positive phenotypically by potentiation disk and were positive for FR-GST genes: 5 harbored
<jats:italic>fosA3</jats:italic>
and 1 harbored
<jats:italic>fosA4</jats:italic>
. To identify the production of FR-GSTs, we set the provisional cutoff value, 5-mm enlargement, by adding PPF to a fosfomycin disk on the MH agar plates containing G6P. Our disk-based potentiation test reliably identifies FR-GST producers and can be performed easily; therefore, it will be advantageous in epidemiological surveys and infection control of fosfomycin-resistant bacteria in clinical settings.
</jats:p>

DOI： 10.1128/JCM.01094-14

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：American Society for Microbiology  

<jats:title>ABSTRACT</jats:title>
<jats:p>
We characterized penicillin-susceptible group B streptococcal (PSGBS) clinical isolates exhibiting no growth inhibition zone around a ceftibuten disk (CTB
<jats:sup>r</jats:sup>
PSGBS). The CTB
<jats:sup>r</jats:sup>
PSGBS isolates, for which augmented MICs of cefaclor and ceftizoxime were found, shared a T394A substitution in penicillin-binding protein 2X (PBP 2X) and a T567I substitution in PBP 2B, together with an additional G429S substitution in PBP 2X or a T145A substitution in PBP 1A, although the T145A substitution in the transglycosidase domain of PBP 1A would have no effect on the level of resistance to ceftibuten.
</jats:p>

DOI： 10.1128/JCM.01291-14

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Vaccine  

The Haemophilus influenzae type b (Hib) vaccine and the heptavalent pneumococcal conjugate vaccine (PCV7) were introduced in Japan in 2008 and 2010, respectively. In 2011, immunization with these two vaccines was encouraged throughout Japan through a governmental program. Children treated in Chiba prefecture for culture-proven invasive H. influenzae disease (IHiD) and invasive Streptococcus pneumoniae disease (IPD) were identified in a prefectural surveillance study from 2008 to 2013. The incidence rate ratio (IRR) and its confidence interval (CI) were calculated to compare the 3 years before and after governmental financial support for vaccination. The average number of IHiD and IPD cases among children <5 years of age in 2011-2013 decreased 84% (IRR: 0.16, 95% CI: 0.09-0.26, p < 0.0001) and 51% (IRR: 0.49, 95% CI: 0.37-0.63, p < 0.0001) compared with those occurring in 2008-2010. The most common non-PCV7 serotype encountered in 2011 and 2013 was 19A. After governmental subsidization of Hib and PCV7 vaccination, IHiD and IPD decreased in Chiba prefecture, Japan. Continuous surveillance is necessary to determine the effectiveness of these two vaccines and for detection of emerging invasive serotypes.

DOI： 10.1016/j.vaccine.2014.07.100

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1002/ams2.64

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Infectious Diseases  

In March 2012, two patients were transported urgently to the hospital in Tottori Prefecture, Japan, because of symptoms suggestive of botulism. Botulinum neurotoxin type A was detected in the clinical specimens and the food consumed by the two patients (vacuum packed adzuki-batto, a sweet adzuki bean soup containing noodles). We were able to make a prompt diagnosis of food botulism associated with the consumption of adzuki-batto, from which the causative pathogen Clostridium botulinum Ab was cultured. © 2014 The Authors.

DOI： 10.1016/j.ijid.2014.01.014

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Clinical Microbiology  

We isolated and characterized three multidrug-resistant clinical isolates of group B streptococci with reduced penicillin susceptibility (PRGBS) that formed small non-beta-hemolytic colonies on sheep blood agar plates but grew well on chocolate agar plates. They can be overlooked in the bacterial identification step, leading to clinical misdiagnosis and treatment failure. © 2014 American Society for Microbiology. All Rights Reserved.

DOI： 10.1128/JCM.00226-14

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Microbiology Society  

<jats:p>We performed a comparative molecular analysis on three types of clinically isolated <jats:italic>Acinetobacter</jats:italic> spp.: epidemic sequence types (STs) of <jats:italic>Acinetobacter baumannii</jats:italic> (epidemic ST-AB), non-epidemic sequence types of <jats:italic>A. baumannii</jats:italic> (non-epidemic ST-AB) and non-<jats:italic>baumannii Acinetobacter</jats:italic> spp. A total of 87 isolates – 46 <jats:italic>A. baumannii</jats:italic>, 25 <jats:italic>A. pittii</jats:italic> and 16 <jats:italic>A. nosocomialis</jats:italic> – from 43 hospitals were analysed. Of these, 31 <jats:italic>A. baumannii</jats:italic> isolates were ST1 or ST2 according to the Pasteur Institute multilocus sequence typing scheme and were defined as epidemic ST-AB. The other 15 <jats:italic>A. baumannii</jats:italic> isolates were defined as non-epidemic ST-AB. The epidemic ST-AB isolates harboured the <jats:italic>bla</jats:italic>
<jats:sub>OXA-23-like</jats:sub> gene or had an IS<jats:italic>Aba1</jats:italic> element upstream of <jats:italic>bla</jats:italic>
<jats:sub>OXA-51-like</jats:sub>, or both, whereas non-epidemic ST-AB and non-<jats:italic>baumannii Acinetobacter</jats:italic> spp. isolates harboured <jats:italic>bla</jats:italic>
<jats:sub>OXA-58-like</jats:sub> or metallo-β-lactamase genes, or both. The proportion of multidrug-resistant isolates was significantly higher in the epidemic ST-AB isolates (48 %) than that in the other types of <jats:italic>Acinetobacter</jats:italic> isolates (5 %) (<jats:italic>P</jats:italic><0.05). In addition, epidemic ST-AB isolates exhibited a relatively higher proportion of fluoroquinolone resistance. We demonstrated that, in terms of genotypes and phenotypes of antimicrobial resistance, non-epidemic ST-AB isolates shared more similarity with non-<jats:italic>baumannii Acinetobacter</jats:italic> spp. isolates than with epidemic ST-AB isolates, regardless of bacterial species. In addition, this study revealed that, even in Japan, where IMP-type metallo-β-lactamase producers are endemic, epidemic ST-AB harbouring <jats:italic>bla</jats:italic>
<jats:sub>IMP</jats:sub> have not yet emerged.</jats:p>

DOI： 10.1099/jmm.0.069138-0

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Antimicrobial Chemotherapy  

DOI： 10.1093/jac/dkt498

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Journal of Infection and Chemotherapy  

Chromobacterium violaceum is sensitive to temperature and the infection is usually confined to tropical or subtropical regions. Since Japan has a warm climate, C. violaceum has been scarcely isolated from clinical specimens. With global warming, however, the geographical distribution of C. violaceum infection is likely to change. We report two cases of C. violaceum nosocomial pneumonia that occurred at an intensive care center in Japan. C. violaceum was first detected from a patient in the same center as a pathogenic organism of pneumonia. Later, the organism was isolated from sputum and a ventilator circuit tube of another patient in the center. The two patients were admitted to the center in nearby beds for several days. All of the pathogens were confirmed to be C. violaceum by the nucleic acid sequence of the 16S rRNA gene and were proven to be genetically identical organisms by pulsed field gel electrophoresis. Both patients were managed with well-humidified and heated oxygen using a venturi mask and ventilator to promote excretion of sputum. It was thought that the medical respiratory care devices that provide a humid and warm environment, an optimal condition for proliferation of C. violaceum, can contribute to C. violaceum infection in a hospital environment. © 2013, Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases.

DOI： 10.1016/j.jiac.2013.10.001

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：1  

DOI： 10.7883/yoken.67.66

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担当区分：筆頭著者,　最終著者,　責任著者   掲載種別：研究論文（学術雑誌）  

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：PLoS ONE