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Antibiotic susceptibility and genome analysis of Enterococcus species isolated from inpatients in one hospital with no apparent outbreak of vancomycin-resistant Enterococcus in Japan 日本一家未明显爆发耐万古霉素肠球菌疫情的医院从住院病人中分离的肠球菌的抗生素敏感性和基因组分析。
IF 1.9 4区 医学 Q4 IMMUNOLOGY Pub Date : 2024-06-14 DOI: 10.1111/1348-0421.13155
Ayumi Fujii, Miki Kawada-Matsuo, Mi Nguyen-Tra Le, Kanako Masuda, Kayoko Tadera, Yujin Suzuki, Saki Nishihama, Junzo Hisatsune, Yo Sugawara, Seiya Kashiyama, Hideki Shiba, Tomonao Aikawa, Hiroki Ohge, Motoyuki Sugai, Hitoshi Komatsuzawa

To prevent nosocomial infection, it is important to screen for potential vancomycin-resistant Enterococcus (VRE) among patients. In this study, we analyzed enterococcal isolates from inpatients in one hospital without any apparent outbreak of VRE. Enterococcal isolates were collected from inpatients at Hiroshima University Hospital from April 1 to June 30, 2021 using selective medium for Enterococci. Multilocus sequence typing, antimicrobial susceptibility testing, and whole-genome sequencing were performed. A total of 164 isolates, including Enterococcus faecium (41 isolates), Enterococcus faecalis (80 isolates), Enterococcus raffinosus (11 isolates), Enterococcus casseliflavus (nine isolates), Enterococcus avium (12 isolates), Enterococcus lactis (eight isolates), Enterococcus gallinarum (two isolates), and Enterococcus malodoratus (one isolate), were analyzed. We found one vanA-positive E. faecium, which was already informed when the patient was transferred to the hospital, nine vanC-positive E. casseliflavus, and two vanC-positive E. gallinarum. E. faecium isolates showed resistance to ampicillin (95.1%), imipenem (95.1%), and levofloxacin (87.8%), and E. faecalis isolates showed resistance to minocycline (49.4%). Ampicillin- and levofloxacin-resistant E. faecium had multiple mutations in penicillin-binding protein 5 (PBP5) (39/39 isolates) and ParC/GyrA (21/36 isolates), respectively. E. raffinosus showed resistance to ampicillin (81.8%), imipenem (45.5%), and levofloxacin (45.5%), and E. lactis showed resistance to ampicillin (37.5%) and imipenem (50.0%). The linezolid resistance genes optrA and cfr(B) were found only in one isolate of E. faecalis and E. raffinosus, respectively. This study, showing the status of enterococci infection in hospitalized patients, is one of the important information when considering nosocomial infection control of VRE.

为预防院内感染,必须筛查患者中可能存在的耐万古霉素肠球菌(VRE)。在本研究中,我们分析了一家未明显爆发 VRE 的医院中住院患者的肠球菌分离物。我们使用肠球菌选择性培养基收集了广岛大学医院 2021 年 4 月 1 日至 6 月 30 日住院患者的肠球菌分离物。进行了多焦点序列分型、抗菌药物药敏试验和全基因组测序。共分析了 164 个分离株,包括粪肠球菌(41 个分离株)、粪肠球菌(80 个分离株)、拉菲诺斯肠球菌(11 个分离株)、卡氏肠球菌(9 个分离株)、阿维氏肠球菌(12 个分离株)、乳酸肠球菌(8 个分离株)、加里纳氏肠球菌(2 个分离株)和恶臭肠球菌(1 个分离株)。我们发现了 1 株 vanA 阳性的粪肠球菌(患者转院时已获知)、9 株 vanC 阳性的卡氏肠球菌和 2 株 vanC 阳性的加里纳氏肠球菌。分离出的粪肠球菌对氨苄西林(95.1%)、亚胺培南(95.1%)和左氧氟沙星(87.8%)具有耐药性,对米诺环素(49.4%)具有耐药性。对氨苄西林和左氧氟沙星耐药的粪肠球菌分别对青霉素结合蛋白 5(PBP5)(39/39 个分离株)和 ParC/GyrA (21/36 个分离株)有多重突变。拉菲诺斯大肠杆菌对氨苄西林(81.8%)、亚胺培南(45.5%)和左氧氟沙星(45.5%)产生耐药性,乳酸杆菌对氨苄西林(37.5%)和亚胺培南(50.0%)产生耐药性。仅在粪大肠杆菌和拉菲诺斯大肠杆菌的一个分离株中分别发现了利奈唑胺耐药基因 optrA 和 cfr(B)。这项研究显示了住院患者肠球菌感染的状况,是考虑控制 VRE 的院内感染时的重要信息之一。
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引用次数: 0
Issue Information – Cover 发行信息 - 封面
IF 2.6 4区 医学 Q3 Immunology and Microbiology Pub Date : 2024-06-05 DOI: 10.1111/1348-0421.13158

Cover photograph: LEfSe analysis of fecal microbiota. Microbiol Immunol: 68:206-211. Article link here

封面照片:粪便微生物群的 LEfSe 分析。微生物学免疫学》:68:206-211。文章链接
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引用次数: 0
Involvement of SARS-CoV-2 accessory proteins in immunopathogenesis SARS-CoV-2 辅助蛋白参与免疫发病机制。
IF 1.9 4区 医学 Q4 IMMUNOLOGY Pub Date : 2024-06-04 DOI: 10.1111/1348-0421.13157
Hayato Ito, Tomokazu Tamura, Lei Wang, Kento Mori, Masumi Tsuda, Rigel Suzuki, Saori Suzuki, Kumiko Yoshimatsu, Shinya Tanaka, Takasuke Fukuhara

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the largest single-stranded RNA virus known to date. Its genome contains multiple accessory protein genes that act against host immune responses but are not required for progeny virus production. The functions of the accessory proteins in the viral life cycle have been examined, but their involvement in viral pathogenicity remains unclear. Here, we investigated the roles of the accessory proteins in viral immunopathogenicity. To this end, recombinant SARS-CoV-2 possessing nonsense mutations in the seven accessory protein open reading frames (ORFs) (ORF3a, ORF3b, ORF6, ORF7a, ORF8, ORF9b, and ORF10) was de novo generated using an early pandemic SARS-CoV-2 strain as a backbone. We confirmed that the resultant virus (termed ORF3–10 KO) did not express accessory proteins in infected cells and retained the desired mutations in the viral genome. In cell culture, the ORF3–10 KO virus exhibited similar virus growth kinetics as the parental virus. In hamsters, ORF3–10 KO virus infection resulted in mild weight loss and reduced viral replication in the oral cavity and lung tissue. ORF3–10 KO virus infection led to mild inflammation, indicating that an inability to evade innate immune sensing because of a lack of accessory proteins impairs virus growth in vivo and results in quick elimination from the body. Overall, we showed that SARS-CoV-2 accessory proteins are involved in immunopathogenicity.

严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)是迄今已知的最大的单链 RNA 病毒。它的基因组中含有多种辅助蛋白基因,这些基因能对抗宿主的免疫反应,但并不是产生后代病毒所必需的。人们已经研究了附属蛋白在病毒生命周期中的功能,但它们与病毒致病性的关系仍不清楚。在此,我们研究了附属蛋白在病毒免疫致病性中的作用。为此,我们以早期大流行的 SARS-CoV-2 株系为骨干,从头生成了在七个附属蛋白开放阅读框(ORF)(ORF3a、ORF3b、ORF6、ORF7a、ORF8、ORF9b 和 ORF10)中具有无义突变的重组 SARS-CoV-2 病毒。我们证实,产生的病毒(称为 ORF3-10 KO)在感染细胞中不表达附属蛋白,并在病毒基因组中保留了所需的突变。在细胞培养中,ORF3-10 KO 病毒表现出与亲本病毒相似的病毒生长动力学。仓鼠感染 ORF3-10 KO 病毒后体重轻微下降,口腔和肺组织中的病毒复制减少。ORF3-10 KO病毒感染会导致轻微炎症,这表明由于缺乏附属蛋白而无法躲避先天性免疫感应会影响病毒在体内的生长,并导致病毒被迅速排出体外。总之,我们发现 SARS-CoV-2 辅助蛋白参与了免疫致病性。
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引用次数: 0
Biofilm-derived membrane vesicles exhibit potent immunomodulatory activity in Pseudomonas aeruginosa PAO1 生物膜衍生的膜囊泡对铜绿假单胞菌 PAO1 表现出强大的免疫调节活性。
IF 1.9 4区 医学 Q4 IMMUNOLOGY Pub Date : 2024-05-26 DOI: 10.1111/1348-0421.13156
Minato Takahara, Satoru Hirayama, Hiroyuki Futamata, Ryoma Nakao, Yosuke Tashiro

Pathogenic bacteria form biofilms on epithelial cells, and most bacterial biofilms show increased production of membrane vesicles (MVs), also known as outer membrane vesicles in Gram-negative bacteria. Numerous studies have investigated the MVs released under planktonic conditions; however, the impact of MVs released from biofilms on immune responses remains unclear. This study aimed to investigate the characteristics and immunomodulatory activity of MVs obtained from both planktonic and biofilm cultures of Pseudomonas aeruginosa PAO1. The innate immune responses of macrophages to planktonic-derived MVs (p-MVs) and biofilm-derived MVs (b-MVs) were investigated by measuring the mRNA expression of proinflammatory cytokines. Our results showed that b-MVs induced a higher expression of inflammatory cytokines, including Il1b, Il6, and Il12p40, than p-MVs. The mRNA expression levels of Toll-like receptor 4 (Tlr4) differed between the two types of MVs, but not Tlr2. Polymyxin B significantly neutralized b-MV-mediated cytokine induction, suggesting that lipopolysaccharide of native b-MVs is the origin of the immune response. In addition, heat-treated or homogenized b-MVs induced the mRNA expression of cytokines, including Tnfa, Il1b, Il6, and Il12p40. Heat treatment of MVs led to increased expression of Tlr2 but not Tlr4, suggesting that TLR2 ligands play a role in detecting the pathogen-associated molecular patterns in lysed MVs. Taken together, our data indicate that potent immunomodulatory MVs are produced in P. aeruginosa biofilms and that this behavior could be a strategy for the bacteria to infect host cells. Furthermore, our findings would contribute to developing novel vaccines using MVs.

致病细菌会在上皮细胞上形成生物膜,大多数细菌生物膜都会增加膜囊(MVs)的产生,革兰氏阴性细菌的膜囊也称为外膜囊。许多研究对浮游生物条件下释放的膜囊泡进行了调查,但生物膜释放的膜囊泡对免疫反应的影响仍不清楚。本研究旨在调查从铜绿假单胞菌 PAO1 的浮游生物和生物膜培养物中获得的 MVs 的特征和免疫调节活性。通过测量促炎细胞因子的 mRNA 表达,研究了巨噬细胞对浮游生物源 MVs(p-MVs)和生物膜源 MVs(b-MVs)的先天性免疫反应。结果显示,与 p-MVs 相比,b-MVs 诱导的炎性细胞因子(包括 Il1b、Il6 和 Il12p40)表达量更高。两种中毒性病毒的 Toll 样受体 4(Tlr4)的 mRNA 表达水平不同,但 Tlr2 的表达水平没有差异。多粘菌素 B 能明显中和 b-MV 介导的细胞因子诱导,这表明原生 b-MV 的脂多糖是免疫反应的起源。此外,热处理或均质化的b-MV可诱导细胞因子(包括Tnfa、Il1b、Il6和Il12p40)的mRNA表达。热处理中毒性细胞可导致 Tlr2 的表达增加,但 Tlr4 的表达却没有增加,这表明 TLR2 配体在检测裂解中毒性细胞中的病原体相关分子模式方面发挥了作用。总之,我们的数据表明,铜绿微囊藻生物膜中会产生强效的免疫调节MV,这种行为可能是细菌感染宿主细胞的一种策略。此外,我们的研究结果将有助于利用 MVs 开发新型疫苗。
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引用次数: 0
The prevention effect of Limosilactobacillus reuteri on acute kidney injury by regulating gut microbiota 通过调节肠道微生物群来预防急性肾损伤的Limosilactobacillus reuteri。
IF 1.9 4区 医学 Q4 IMMUNOLOGY Pub Date : 2024-05-15 DOI: 10.1111/1348-0421.13130
Zhan Yang, Juan Ni, Xuewei Sun, Qian Cui, Xinrui Zhang, Mingyan Zhang, Xiaojing Zhu, Zihan Wu, Chengliang Tang, Jingfeng Zhu, Huijuan Mao, Kang Liu, Chunhui Wang, Changying Xing, Jin Zhu

Acute kidney injury (AKI) has considerably high morbidity and mortality but we do not have proper treatment for it. There is an urgent need to develop new prevention or treatment methods. Gut microbiota has a close connection with renal diseases and has become the new therapy target for AKI. In this study, we found the oral administration of the probiotic Limosilactobacillus reuteri had a prevention effect on the AKI induced by lipopolysaccharide (LPS). It reduced serum concentration of creatinine and urea nitrogen and protected the renal cells from necrosis and apoptosis. Meanwhile, L. reuteri improved the gut barrier function, which is destroyed in AKI, and modulated the gut microbiota and relevant metabolites. Compared with the LPS group, L. reuteri increased the proportion of Proteobacteria and reduced the proportion of Firmicutes, changing the overall structure of the gut microbiota. It also influenced the fecal metabolites and changed the metabolite pathways, such as tyrosine metabolism, pentose and glucuronate interconversions, galactose metabolism, purine metabolism, and insulin resistance. These results showed that L. reuteri is a potential therapy for AKI as it helps in sustaining the gut barrier integrity and modulating gut microbiota and related metabolites.

急性肾损伤(AKI)具有相当高的发病率和死亡率,但我们却没有适当的治疗方法。我们迫切需要开发新的预防或治疗方法。肠道微生物群与肾脏疾病关系密切,已成为治疗急性肾损伤的新靶点。本研究发现,口服益生菌Limosilactobacillus reuteri对脂多糖(LPS)诱导的AKI有预防作用。它能降低血清肌酐和尿素氮的浓度,保护肾细胞免受坏死和凋亡。同时,L. reuteri 还能改善 AKI 中被破坏的肠道屏障功能,调节肠道微生物群和相关代谢物。与 LPS 组相比,L. reuteri 增加了蛋白菌的比例,降低了固缩菌的比例,改变了肠道微生物群的整体结构。它还影响了粪便代谢物,改变了代谢物的途径,如酪氨酸代谢、戊糖和葡萄糖醛酸的相互转化、半乳糖代谢、嘌呤代谢和胰岛素抵抗。这些结果表明,L. reuteri 是治疗 AKI 的一种潜在疗法,因为它有助于维持肠道屏障的完整性,调节肠道微生物群和相关代谢物。
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引用次数: 0
Issue Information – Cover 发行信息 - 封面
IF 2.6 4区 医学 Q3 Immunology and Microbiology Pub Date : 2024-05-06 DOI: 10.1111/1348-0421.13129

Cover photograph: Proposed model: Mutations in N-glycosylation sites at N331 and N343 residues of RBD 705 protein inhibit S-ACE2 binding, IL-6 expression and cytotoxicity. Microbiol Immunol: 68:165-178. Article link here

封面照片:拟议模型:RBD 705 蛋白 N331 和 N343 残基的 N-糖基化位点突变抑制 S-ACE2 结合、IL-6 表达和细胞毒性。微生物学免疫学》:68:165-178。文章链接
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引用次数: 0
Possible link between colonization of the gastrointestinal tract by Citrobacter rodentium in C57BL/6 mice and microbiota composition C57BL/6 小鼠胃肠道啮齿动物柠檬杆菌定植与微生物群组成之间的可能联系。
IF 2.6 4区 医学 Q3 Immunology and Microbiology Pub Date : 2024-04-21 DOI: 10.1111/1348-0421.13128
Tsuyoshi Miki, Takeshi Haneda, Nobuhiko Okada, Masahiro Ito

Colonization resistance, conferred by the host's microbiota through both direct and indirect protective actions, serves to protect the host from enteric infections. Here, we identified the specific members of the gut microbiota that impact gastrointestinal colonization by Citrobacter rodentium, a murine pathogen causing colonic crypt hyperplasia. The gut colonization levels of C. rodentium in C57BL/6 mice varied among breeding facilities, probably due to differences in microbiota composition. A comprehensive analysis of the microbiota revealed that specific members of the microbiota may influence gut colonization by C. rodentium, thus providing a potential link between the two.

宿主的微生物群通过直接和间接的保护作用赋予宿主定植抗性,从而保护宿主免受肠道感染。在这里,我们确定了肠道微生物群中影响棒状杆菌胃肠道定植的特定成员,棒状杆菌是一种导致结肠隐窝增生的小鼠病原体。可能由于微生物群组成的差异,C57BL/6小鼠的肠道鼠杆菌定植水平在不同饲养设施中有所不同。对微生物群的综合分析表明,微生物群的特定成员可能会影响鼠疫杆菌的肠道定植,从而提供了两者之间的潜在联系。
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引用次数: 0
Proposal of Helicobacter higonensis sp. nov. isolated from a human clinical specimen, and emended description of Helicobacter valdiviensis Collado, 2014 从人类临床标本中分离出新的Helicobacter higonensis sp.的建议,以及对Helicobacter valdiviensis Collado, 2014的修订描述
IF 2.6 4区 医学 Q3 Immunology and Microbiology Pub Date : 2024-04-10 DOI: 10.1111/1348-0421.13127
Junko Tomida, Tohru Miyoshi-Akiyama, Ryo Kutsuna, Hiroyasu Tsutsuki, Tomohiro Sawa, Margo Cnockaert, Peter Vandamme, Yoshiaki Kawamura

We have previously isolated a gram-negative microaerophilic strain, PAGU2000T from a patient presenting with a fever in Kumamoto Prefecture, Japan. The present study aimed to comprehensively analyze the taxonomy of the isolated strain using a polyphasic approach. The 16S rRNA gene sequence analysis indicated that the strain was a member of enterohepatic Helicobacter. The strain PAGU2000T shared a 97.5% 16S rRNA gene nucleotide identity with Helicobacter valdiviensis, and this taxonomic position was confirmed by phylogenetic analysis of the GyrA amino acid sequences. The proposed strain PAGU2000T has a 1.482 Mbp chromosome with a DNA G + C content of 31.3 mol% and encodes 1520 predicted coding sequences. The average nucleotide identity between the strain PAGU2000T and type strain of H. valdiviensis was 70.3%, which was lower than the recommended threshold of 95% for species delineation. The strain PAGU2000T was a motile, non-spore-forming, and spiral-shaped bacterium, exhibiting catalase and oxidase activities but not urease and nitrate reduction. This study demonstrates that the isolate represents a novel species within enterohepatic Helicobacter, for which the name Helicobacter higonensis is proposed (type strain: PAGU2000T = GTC 16811T = LMG 33095T). In this study, we describe the phenotypic and morphological features of this strain and propose an emended description of some biochemical traits of H. valdiviensis.

我们曾从日本熊本县的一名发烧患者身上分离出一株嗜微气的革兰氏阴性菌 PAGU2000T。本研究旨在采用多相法全面分析分离菌株的分类。16S rRNA 基因序列分析表明,该菌株属于肠肝螺旋杆菌。菌株 PAGU2000T 与瓦尔迪维氏螺旋杆菌的 16S rRNA 基因核苷酸相同度为 97.5%,GyrA 氨基酸序列的系统进化分析证实了这一分类学定位。拟议的菌株 PAGU2000T 的染色体为 1.482 Mbp,DNA G + C 含量为 31.3 mol%,编码 1520 个预测的编码序列。菌株 PAGU2000T 与 H. valdiviensis 型菌株的平均核苷酸同一性为 70.3%,低于建议的 95% 的物种划分阈值。菌株 PAGU2000T 是一种能运动、不形成孢子、呈螺旋状的细菌,具有过氧化氢酶和氧化酶活性,但不具有脲酶和硝酸盐还原活性。这项研究表明,该分离菌株代表了肠肝螺旋杆菌中的一个新物种,并将其命名为希贡螺旋杆菌(模式菌株:PAGU2000T = GTC 16811T = LMG 33095T)。在本研究中,我们描述了该菌株的表型和形态特征,并提出了对 H. valdiviensis 某些生化特征的修正描述。
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引用次数: 0
Issue Information – Cover 发行信息 - 封面
IF 2.6 4区 医学 Q3 Immunology and Microbiology Pub Date : 2024-04-09 DOI: 10.1111/1348-0421.13126

Cover photograph: Streptococcus pneumoniae ΔlspA strain induces less SEAP production in THP-1-Blue cells. Microbiol Immunol: 68:155-159. Article link here

封面照片:肺炎链球菌 ΔlspA 菌株诱导 THP-1-Blue 细胞产生较少的 SEAP。微生物学免疫学》:68:155-159。文章链接
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引用次数: 0
Issue Information – Cover 发行信息 - 封面
IF 2.6 4区 医学 Q3 Immunology and Microbiology Pub Date : 2024-03-11 DOI: 10.1111/1348-0421.13125

Cover photograph: The modulatory function of circ_0008410 was examined in RASFs. The amplification of divergent primers and 518 convergent primer in cDNA and gDNA of RASFs. Microbiol Immunol: 68:100-110. Article link here

封面照片:研究了 circ_0008410 在 RASFs 中的调节功能。在 RASFs 的 cDNA 和 gDNA 中扩增分歧引物和 518 融合引物。微生物学免疫学》:68:100-110。文章链接
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引用次数: 0
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