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103. Treatment history shapes the evolution of complex carbapenem-resistant phenotypes in Klebsiella spp.

Author

Rosas, Natalia C., primary, Wilksch, Jonathan J., additional, Barber, Jake, additional, Li, Jiahui, additional, Wang, Yanan, additional, Sun, Zhewei, additional, Rocker, Andrea, additional, Webb, Chaille, additional, Perlaza-Jiménez, Laura, additional, Stubenrauch, Christopher, additional, Vijaykrishna, Dhanasekaran, additional, Song, Jiangning, additional, Taiaroa, George, additional, Davies, Mark, additional, Strugnell, Richard A., additional, Bao, Qiyu, additional, Zhou, Tieli, additional, McDonald, Michael J., additional, and Lithgow, Trevor, additional

Published
2022
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107. Acquisition of Daptomycin Resistance by Enterococcus faecium Confers Collateral Sensitivity to Glycopeptides

Author

Zeng, Weiliang, Feng, Luozhu, Qian, Changrui, Chen, Tao, Wang, Sipei, Zhang, Ying, Zheng, Xiangkuo, Wang, Lingbo, Liu, Shixing, Zhou, Tieli, and Sun, Yao

Subjects
Microbiology (medical), biochemical phenomena, metabolism, and nutrition, bacterial infections and mycoses, Microbiology
Abstract

Daptomycin is a last-line antibiotic used in the treatment of multidrug-resistant Enterococcus faecium infections. Alarmingly, daptomycin-resistant E. faecium isolates have emerged. In this study, we investigated the evolution and mechanisms of daptomycin resistance in clinical E. faecium isolates and the corresponding acquisition of collateral sensitivity (CS) as an evolutionary trade-off. We evolved daptomycin resistance in six daptomycin-susceptible E. faecium isolates to obtain daptomycin-resistant mutants. The six E. faecium strains successfully acquired high-level resistance to daptomycin in vitro, but this led to fitness costs in terms of growth, in vitro competition, and virulence. Mutations in liaFSR, yycFG, and cls; increased surface positive charge; thicker cell walls; and elevated expression of dltABCD and tagGH were observed in daptomycin-resistant mutants. Surprisingly, we observed the emergence of CS in SC1762 isolates after the induction of daptomycin resistance. Compared with parental strains, the SC1174-D strain (i.e., daptomycin-resistant mutant of SC1174; non-CS) showed significantly upregulated expression of the vanA gene cluster. However, in SC1762-D (i.e., daptomycin-resistant mutant of SC1762), all vanA cluster genes except the vanX gene were obviously downregulated. Further in silico analyses revealed that an IS1216E-based composite transposon was generated in SC1762-D, and it disrupted the vanH gene, likely affecting the structure and expression of the vanA gene cluster and resulting in resensitization to glycopeptides. Overall, this study reports a novel form of CS between daptomycin and glycopeptides in E. faecium. Further, it provides a valuable foundation for developing effective regimens and sequential combinations of daptomycin and glycopeptides against E. faecium.

Published
2022
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108. Emergence of Ceftazidime–Avibactam Resistance and Decreased Virulence in Carbapenem-Resistant ST11 Klebsiella pneumoniae During Antibiotics Treatment

Author

Xu,Mengxin, Qian,Changrui, Jia,Huaiyu, Feng,Luozhu, Shi,Shiyi, Zhang,Ying, Wang,Lingbo, Cao,Jianming, Zhou,Tieli, Zhou,Cui, Xu,Mengxin, Qian,Changrui, Jia,Huaiyu, Feng,Luozhu, Shi,Shiyi, Zhang,Ying, Wang,Lingbo, Cao,Jianming, Zhou,Tieli, and Zhou,Cui

Abstract

Mengxin Xu,1 Changrui Qian,2 Huaiyu Jia,1 Luozhu Feng,3 Shiyi Shi,1 Ying Zhang,3 Lingbo Wang,1 Jianming Cao,3 Tieli Zhou,1 Cui Zhou1 1Department of Clinical Laboratory, the First Affiliated Hospital of Wenzhou Medical University, Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province, Wenzhou, People’s Republic of China; 2School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, People’s Republic of China; 3Department of Medical Laboratory Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, People’s Republic of ChinaCorrespondence: Tieli Zhou; Cui Zhou, Department of Clinical Laboratory, the First Affiliated Hospital of Wenzhou Medical University; Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province, Wenzhou, 325035, People’s Republic of China, Tel +86 0577 8668 9885, Email wyztli@163.com; ZHOUCUI0826@163.comIntroduction: Carbapenem-resistant Klebsiella pneumoniae (CRKP) poses a serious threat to human public health. Ceftazidime–avibactam (CZA) is currently one of the few effective antibiotics for carbapenem-resistant Enterobacteriaceae (CRE).Methods and Results: Here, we analyzed two longitudinal Klebsiella pneumoniae clinical isolates (FK8578, FK8695) that were isolated from an ICU patient during antimicrobial treatment. Broth microdilution method, whole-genome sequencing (WGS) and comparative genomic analysis were used to elucidate the dynamics and mechanisms of antibiotic resistance. String test, quantification of capsule, biofilm inhibition test and Galleria mellonella (G. mellonella) infection model were used to explore the changes in virulence of the two clinical isolates. During antibiotic treatment, CRKP FK8578 underwent a series of drug resistance and virulence changes, including CZA resistance, carbapenem susceptibility and virulence attenuation. The results of WGS showed that mutation of blaKPC-2 to blaKPC-3

Published
2022

109. Novel Insight of Transcription Factor PtrA on Pathogenicity and Carbapenems Resistance in Pseudomonas aeruginosa

Author

Zhang,Ying, Wang,Lingbo, Chen,Liqiong, Zhu,Peiwu, Huang,Na, Chen,Tao, Chen,Lijiang, Wang,Zhongyong, Liao,Wenli, Cao,Jianming, Zhou,Tieli, Zhang,Ying, Wang,Lingbo, Chen,Liqiong, Zhu,Peiwu, Huang,Na, Chen,Tao, Chen,Lijiang, Wang,Zhongyong, Liao,Wenli, Cao,Jianming, and Zhou,Tieli

Abstract

Ying Zhang,1,* Lingbo Wang,1,* Liqiong Chen,2 Peiwu Zhu,1 Na Huang,1 Tao Chen,1 Lijiang Chen,1 Zhongyong Wang,1 Wenli Liao,1 Jianming Cao,2 Tieli Zhou1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province, Wenzhou, People’s Republic of China; 2Department of Medical Laboratory Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Tieli Zhou, Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province, Wenzhou, 325035, People’s Republic of China, Tel +86-577-8668-9885, Email wyztli@163.com Jianming Cao, Department of Medical Laboratory Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, 325035, People’s Republic of China, Tel +86-577-88069595, Email wzcjming@163.comIntroduction: Globally, Pseudomonas aeruginosa (PA) is emerging as a predominant nosocomial pathogen that often induces aggressive and even deadly infections. Pseudomonas type III repressor A (PtrA) can be activated specifically by copper ions and interacts with type-III transcriptional activator ExsA. This study aims to provide insight into the PtrA-mediated regulation of the pathogenicity and antibiotics resistance of PA.Methods and Results: The results of transcriptome sequencing analyses and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) showed that PtrA plays a dual regulatory role in the virulence systems of PA: negatively regulates the type-III secretion system (T3SS) and positively regulates the quorum-sensing system (QS). The ptrA mutant attenuated extracellular virulence related to QS like pyocyanin, elas

Published
2022

116. Mechanisms of Heteroresistance and Resistance to Imipenem in Pseudomonas aeruginosa

Author

Xu,Ye, Zheng,Xiangkuo, Zeng,Weiliang, Chen,Tao, Liao,Wenli, Qian,Jiao, Lin,Jie, Zhou,Cui, Tian,Xuebin, Cao,Jianming, and Zhou,Tieli

Subjects
resistance, Infection and Drug Resistance, Pseudomonas aeruginosa, oprD, heteroresistance, molecular mechanism, Original Research, imipenem
Abstract

Ye Xu,1 Xiangkuo Zheng,2 Weiliang Zeng,2 Tao Chen,1 Wenli Liao,1 Jiao Qian,1 Jie Lin,1 Cui Zhou,1 Xuebin Tian,2 Jianming Cao,2 Tieli Zhou1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 2Department of Medical Laboratory Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaCorrespondence: Jianming Cao; Tieli Zhou Tel +86-577-88069595; +86-577-86689885Email wzcjming@163.com; wyztli@163.comBackground: Heteroresistance is a phenomenon that occurs in all bacteria and can cause treatment failure. Yet, the exact mechanisms responsible for heteroresistance still remain unknown. The following study investigated the mechanisms of imipenem-heteroresistance and -resistance in Pseudomonas aeruginosa clinical isolates from Wenzhou, China.Methods: Imipenem resistance was detected by the agar dilution method; heteroresistance was determined by population analysis profiles. Biofilm formation assay and modified carbapenem inactivation methods were also performed. Polymerase chain reaction (PCR) was conducted to detect oprD, and quantitative real-time PCR was used to determine expression levels of oprD, ampC and four efflux pump coding genes (mexB, mexD, mexE and mexY).Results: Six imipenem-heteroresistant and -resistant P. aeruginosa isolates were selected respectively. Deficient oprD was detected in all resistant isolates and two heteroresistant isolates. No strains produced carbapenemases. Expression levels of oprD were down-regulated in heteroresistant isolates. Transcription levels of the mexE and mexY were significantly increased in all heterogeneous subpopulations compared with their respective native ones. Compared with the susceptible group, increased mean relative expression levels of mexE and mexY or the decreased mean relative expression levels of oprD were observed in the resistant group (P < 0.05), whereas transcription levels of the mexB and mexD remained unchanged.Conclusion: Down-regulation of oprD contributed to the resistance and heteroresistance of imipenem in our P. aeruginosa clinical isolates. In addition, the marginal up-regulation of efflux systems may indirectly affect imipenem resistance. Contrarily, defective oprD was less common in our experimental heteroresistant strains than resistant strains.Keywords: Pseudomonas aeruginosa, imipenem, resistance, heteroresistance, molecular mechanism, oprD

Published
2020

117. Dynamic Epidemiology and Virulence Characteristics of Carbapenem-Resistant Klebsiella pneumoniae in Wenzhou, China from 2003 to 2016

Author

Zhao, Yajie, Zhang, Siqin, Fang, Renchi, Wu, Qing, Li, Jiahui, Zhang, Yizhi, Rocker, Andrea, Cao, Jianming, Lithgow, Trevor, and Zhou, Tieli

Subjects
Klebsiella pneumoniae, ST11, Infection and Drug Resistance, carbapenem-resistant, epidemiology, antimicrobial resistance, Original Research
Abstract

Yajie Zhao,1,2 Siqin Zhang,1 Renchi Fang,1 Qing Wu,1 Jiahui Li,1 Yizhi Zhang,1 Andrea Rocker,3 Jianming Cao,2 Trevor Lithgow,3 Tieli Zhou1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 2School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, People’s Republic of China; 3Infection and Immunity Program, Biomedicine Discovery Institute and Department of Microbiology, Monash University, Melbourne, VIC, AustraliaCorrespondence: Trevor Lithgow; Tieli Zhou Email trevor.lithgow@monash.edu; wyztli@163.comPurpose: To investigate transitions in resistance mechanisms, virulence characteristics and molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) during 2003– 2016 in a major Eastern Chinese medical center.Patients and Methods: From a total of 2299 K. pneumoniae clinical strains collected from 2003 to 2016, 214 were found to be CRKP isolates and were selected for further study. Characterization of these was conducted by molecular detection of antibiotic resistance markers and virulence determinants, modified carbapenem inactivation method and multilocus sequence typing (MLST).Results: In this study, the prevalence of CRKP was increasing over the 14-year period, mirroring a national trend. These CRKP strains were resistant to most of the tested, clinicallyrelevant drugs. The majority of these CRKP strains were positive for carbapenemases, with the Klebsiella pneumoniae carbapenemase (KPC) found to be the dominant type (207/210, 98.6%). The carrier rates of virulence genes uge, entB, fimH, mrkD and ureA increased in 2016, while the ybtA, iucAand irp2 showed a relatively constant trend. From MLST data, ST11 (88.8%, 190/214) was the preponderant sequence type (ST), followed by ST15 (1.9%, 4/214) and ST656 (1.4%, 3/214). Several strains with less common STs (ST690, ST895, ST1823 and ST1384) were also detected, and these too showed high levels of antimicrobial resistance.Conclusion: The average national rise in CRKP across China is mirrored in this in-depth analysis of a single hospital, while the prevalence of hypervirulent CRKP (such as ST15) was relatively low as of 2016. Continuous monitoring is necessary to keep track of CRKP and should include the prospect of newly emerging strains with less common STs and the prospect of detecting carbapenem-resistant, carbapenemase-negative Klebsiella pneumoniae.Keywords: Klebsiella pneumoniae, carbapenem-resistant, antimicrobial resistance, ST11, epidemiology

Published
2020

118. Molecular Mechanisms and Epidemiology of Carbapenem-Resistant Escherichia coli Isolated from Chinese Patients During 2002–2017

Author

Tian, Xuebin, Zheng, Xiangkuo, Sun, Yao, Fang, Renchi, Zhang, Siqin, Zhang, Xiucai, Lin, Jie, Cao, Jianming, and Zhou, Tieli

Subjects
carbapenemase, outer membrane porin, carbapenem-resistant, Escherichia coli, bacteria, epidemiology, biochemical phenomena, metabolism, and nutrition, bacterial infections and mycoses, Original Research
Abstract

Background The emergence and spread of carbapenem-resistant Escherichia coli (E. coli) pose a serious threat to human health worldwide. This study aimed to investigate the molecular mechanisms underlying carbapenem resistance and their prevalence among E. coli in China. Methods A collection of 5796 E. coli clinical isolates were collected from the First Affiliated Hospital of Wenzhou Medical University from 2002 to 2017. Sensitivity to antibiotics was determined using the agar dilution method. The detection of carbapenemases production and the prevalence of resistance-associated genes were investigated through modified carbapenem inactivation method (mCIM), PCR and sequencing. The mutations in outer membrane porins genes (ompC and ompF) were also analyzed by PCR and sequencing assays. The effect of efflux pump mechanism on carbapenem resistance was also tested. E. coli were typed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results A total of 58 strains (1.0%) of carbapenem-resistant E. coli were identified. The strains carrying blaKPC-2 and blaNDM accounted for 22.4% (13/58) and 51.7% (30/58), respectively. Among blaNDM- positive strains, 27 blaNDM genes were assigned to blaNDM-5, while the remaining three strains were blaNDM-1, whereas blaVIM, blaIMP, blaOXA-48, and blaSHV were not found. The CTX-M-type β-lactamase genes accounted for 96.6% (56/58). In addition, blaTEM-1 genes were identified in 58.6% of tested strains. In carbapenem-resistant isolates, mutations in OmpC (the majority of mutated sites were D192G and Q104_F141del, accounting for 54.5%) and OmpF (large deletions S75_V127del, W83_D135del and Q88_D135del) were detected. Of note, the antibiotic resistance was not associated with overexpression of efflux pump. Moreover, MLST categorized the 58 carbapenem-resistant isolates into 19 different sequence types. PFGE analysis revealed that homology among the carbapenem-resistant isolates was low and sporadic. Conclusion The blaNDM was the principal resistance mechanism of carbapenem-resistant E. coli in the hospital. blaNDM-5 is becoming a new threat to public health and the alteration of outer membrane porins might help further increase the MIC of carbapenem.

Published
2020

121. High level glucose increases mutagenesis in human lymphoblastoid cells

Author

Ying Zhang, Junqing Zhou, Tieli Wang, Lu Cai

Subjects
Biology (General), QH301-705.5
Abstract

Epidemiological data have suggested an increased cancer rates in diabetic patients, for which the underlying mechanism is poorly understood. We studied whether high level of glucose (HG) treatment that mimic the hyperglycemic condition in diabetes mellitus is mutagenic. Mutagenesis studies were carried out at both hypoxanthine phosphoribosyltransferase (hprt) and thymidine kinase (tk) loci. Role of p53 in HG-induced mutagenesis was also investigated by using human lymphoblastoid cell lines derived from same donor but differs in p53 statuses; TK6 has wild-type p53, NH32 has null p53, and WTK1 has mutant p53 (ile237). In addition, we studied the influence of antioxidant treatment on HG-induced mutagenesis. Mutation fractions at both loci increased significantly in all three lines at 21 and 28 days after HG treatments. At tk locus, the increase of a class of mutants with normal growth rate is mainly responsible for the overall increased mutant fraction. Compared to TK6 cells, both NH32 and WTK1 cells showed an early onset of mutagenesis. Treatment of cells with antioxidant N-acetyl-L-cysteine partially reduced HG induced mutagenesis. This study is the first to indicate that HG is able to induce gene mutation which may be one of the important mechanisms of diabetes-associated carcinogenesis.

Published
2007

122. Evaluation of resazurin microplate method for rapid detection of vancomycin and linezolid resistance in Enterococcus faecalis and Enterococcus faecium clinical isolates.

Author

Zeng, Weiliang, Han, Yijia, Zheng, Xiangkuo, Yao, Zhuocheng, Xu, Chunquan, Zhang, Xiaotuan, Tang, Miran, Shen, Mo, and Zhou, Tieli

Subjects
ENTEROCOCCUS, VANCOMYCIN resistance, ENTEROCOCCUS faecalis, ENTEROCOCCUS faecium, RESAZURIN, LINEZOLID
Abstract

Background Vancomycin and linezolid resistance among enterococci is an increasing problem due to a lack of alternative antibiotics. Early identification of vancomycin-resistant and linezolid-resistant strains can help prevent the spread of resistance to these antibiotics. Hence, early, rapid and accurate detection of vancomycin and linezolid resistance is critical. Objectives The resazurin microplate method (RMM) was developed for detecting vancomycin and linezolid susceptibility among Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) clinical isolates, and its performance was further evaluated. Methods A total of 209 non-duplicate clinical isolates and three strains from the faeces of domestic animals, including 142 E. faecalis (71 linezolid non-susceptible and 71 linezolid susceptible) and 70 E. faecium (23 vancomycin non-susceptible, 23 vancomycin susceptible, 12 linezolid non-susceptible and 12 linezolid susceptible), were tested using RMM. Results The susceptibility of E. faecium to vancomycin was detected within 5 h, with high susceptibility (23/23) and specificity (23/23). The susceptibility of E. faecalis and E. faecium to linezolid was detected within 4 h, with specificities of 98.59% and 100% and susceptibilities of 94.37% and 58.33% for E. faecalis and E. faecium , respectively. Conclusions RMM had a good positive predictive value for the detection of vancomycin-non-susceptible E. faecium and linezolid-non-susceptible E. faecalis. It thus has the potential to become an alternative method for the rapid screening of these resistant pathogens in clinical practice. [ABSTRACT FROM AUTHOR]

Published
2023
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129. A novel transposon Tn7540 carrying blaNDM-9and fosA3in chromosome of a pathogenic multidrug-resistant Salmonella entericaserovar Indiana isolated from human faeces

Author

Sun, Yao, Han, Yijia, Qian, Changrui, Zhang, Qi, Yao, Zhuocheng, Zeng, Weiliang, Zhou, Tieli, and Wang, Zhongyong

Abstract

•First report of chromosome-encoded blaNDM-9in S.Indiana isolated from human.•Multiple resistance genes were carried by a novel transposon Tn7540.•IS26-flanked Tn7540 was transferable with circular intermediate.•S. Indiana QT6365 was closely related to S. Indiana C629 isolated from chicken.

Published
2023
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136. Additional file 2 of Characterization of resistance mechanisms of Enterobacter cloacae Complex co-resistant to carbapenem and colistin

Author

Liu, Shixing, Fang, Renchi, Zhang, Ying, Chen, Lijiang, Huang, Na, Yu, Kaihang, Zhou, Cui, Cao, Jianming, and Zhou, Tieli

Subjects
genetic structures, polycyclic compounds, bacteria, biochemical phenomena, metabolism, and nutrition, bacterial infections and mycoses
Abstract

Additional file 2: Table S2. MICs of carbapenem and colistin to 19 ECC strains.

Published
2021
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137. Molecular Mechanisms and Epidemiology of Carbapenem-Resistant Enterobacter cloacae Complex Isolated from Chinese Patients During 2004–2018

Author

Liu,Shixing, Huang,Na, Zhou,Cui, Lin,Yishuai, Zhang,Ying, Wang,Lingbo, Zheng,Xiangkuo, Zhou,Tieli, Wang,Zhongyong, Liu,Shixing, Huang,Na, Zhou,Cui, Lin,Yishuai, Zhang,Ying, Wang,Lingbo, Zheng,Xiangkuo, Zhou,Tieli, and Wang,Zhongyong

Abstract

Shixing Liu,1 Na Huang,1 Cui Zhou,1 Yishuai Lin,2 Ying Zhang,2 Lingbo Wang,1 Xiangkuo Zheng,1 Tieli Zhou,1 Zhongyong Wang1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 2School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaCorrespondence: Zhongyong Wang; Tieli ZhouDepartment of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaTel +86-577-5557-9725; +86-577-8668-9885Email wangforever2000@163.com; wyztli@163.comBackground: The emergence and spread of carbapenem-resistant Enterobacter cloacae complex (ECC) have posed a serious threat to human health worldwide. This study aimed to investigate the molecular mechanism of carbapenem resistance and its prevalence among ECC in China.Methods: A total of 1314 ECC clinical isolates were collected from the First Affiliated Hospital of Wenzhou Medical University from 2004 to 2018. Sensitivity to antibiotics was determined using the agar dilution method. The production of carbapenemases and the prevalence of resistance-associated genes were investigated using PCR. The expression of outer membrane porin (OMP) genes (ompC/ompF) and cephalosporinase gene ampC was analyzed by quantitative real-time PCR. The effect of efflux pump mechanism on carbapenem resistance was tested. ECC was typed by multilocus sequence typing (MLST).Results: In this study, 113 carbapenem-nonsusceptible ECC strains were identified. The prevalence rates of carbapenemase genes blaKPC-2 and blaNDM were 12.4% (14/113) and 17.7% (20/113), and that of the extended-spectrum β-lactamase (ESBL) genes blaCTX-M, blaTEM, and blaSHV were 28.3% (32/113), 27.4% (31/113), and 14.2% (16/113), respectively. Among 67 carbapenem-nonsusceptible ECC isolates producing non-carbapenemase, low expression of ompC

Published
2021

138. Comparison of Anti-Microbic and Anti-Biofilm Activity Among Tedizolid and Radezolid Against Linezolid-Resistant Enterococcus faecalis Isolates

Author

Wang,Lingbo, Zhang,Ying, Liu,Shixing, Huang,Na, Zeng,Weiliang, Xu,Wenya, Zhou,Tieli, Shen,Mo, Wang,Lingbo, Zhang,Ying, Liu,Shixing, Huang,Na, Zeng,Weiliang, Xu,Wenya, Zhou,Tieli, and Shen,Mo

Abstract

Lingbo Wang,1,&ast; Ying Zhang,2,&ast; Shixing Liu,1 Na Huang,1 Weiliang Zeng,1 Wenya Xu,1 Tieli Zhou,1 Mo Shen1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 2School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Mo Shen; Tieli ZhouDepartment of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of ChinaTel +86-577-8668-9885Email shenmo601@163.com; wyztli@163.comBackground: The emergence and spread of linezolid-resistant Enterococcus faecalis (E. faecalis) have emerged as a serious threat to human health globally. Therefore, this study aims to compare the anti-microbic as well as the anti-biofilm activity of linezolid, tedizolid, and radezolid against linezolid-resistant E. faecalis.Methods: A total of 2128 E. faecalis isolates were assessed from the First Affiliated Hospital of Wenzhou Medical University from 2011 to 2019. Antibiotic sensitivity was evaluated using the micro broth dilution method. Oxazolidinone-resistant chromosomal and plasmid-borne genes such as cfr, cfr(A), cfr(B), cfr(C), cfr(D), optrA, and poxtA were detected by PCR and then sequenced to detect the presence of mutations in the domain V of the 23S rRNA and the ribosomal proteins L3, L4, and L22. Conjugation experiments were conducted using the broth method. The inhibition and eradication of biofilm were evaluated through crystal violet staining, whereas the efflux pump activities were detected by agar dilution.Results: Out of 2128 isolated E. faecalis, 71 (3.34%) were linezolid-resistant isolates in which the MICs of tedizolid and radezolid ranged from 1 to 4 μg/mL and 0.5– 1 μg/mL, respectively. The MIC50/MIC90 of tedizolid and radezolid were 4 and 8-fold lower than th

Published
2021

139. Outbreak of Multidrug-Resistant OXA-232-Producing ST15 Klebsiella pneumoniae in a Teaching Hospital in Wenzhou, China

Author

Jia,Huaiyu, Zhang,Ying, Ye,Jianzhong, Xu,Wenya, Xu,Ye, Zeng,Weiliang, Liao,Wenli, Chen,Tao, Cao,Jianming, Wu,Qing, Zhou,Tieli, Jia,Huaiyu, Zhang,Ying, Ye,Jianzhong, Xu,Wenya, Xu,Ye, Zeng,Weiliang, Liao,Wenli, Chen,Tao, Cao,Jianming, Wu,Qing, and Zhou,Tieli

Abstract

Huaiyu Jia,1 Ying Zhang,2 Jianzhong Ye,1 Wenya Xu,1 Ye Xu,1 Weiliang Zeng,2 Wenli Liao,1 Tao Chen,1 Jianming Cao,2 Qing Wu,1 Tieli Zhou1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 2Department of Medical Laboratory Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaCorrespondence: Tieli Zhou; Qing WuDepartment of Clinical Laboratory, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of ChinaTel/Fax +86-0577-8668-9885Email wyztli@163.com; wuqing830@163.comBackground: OXA-232-producing carbapenem-resistant Klebsiella pneumoniae (CRKP) has the potential to become the “third epidemic” of carbapenem-resistant Klebsiella strain after KPC-2 and NDM in China. We investigated the first outbreak of CRKP in the First Affiliated Hospital of Wenzhou Medical University.Methods: We collected 610 clinical isolates of CRKP from the First Affiliated Hospital of Wenzhou Medical University between January 2019 and September 2020 and screened them by Polymerase Chain Reaction (PCR). The multilocus sequence typing and pulsed-field gel electrophoresis were used to determine the genetic relatedness of the strains. The antimicrobial susceptibility test was performed to determine the drug resistance of the clinical isolates. The molecular mechanism underlying carbapenem resistance was elucidated by performing PCR and conjugation experiments. The virulence potential of the strains was determined by the string test, detection of virulence-associated genes and capsular serotypes, and Galleria mellonella larval infection model.Results: Between September 2019 and May 2020, 26 OXA-232-producing CRKP were obtained from 12 patients in our hospital. Ten patients were hospitalized in the intensive care units (ICU) and the overall mortality of the inpatients inv

Published
2021

140. In vitro Activity of Meropenem-Vaborbactam versus Other Antibiotics Against Carbapenem-Resistant Escherichia coli from Southeastern China

Author

Huang,Na, Chen,Tao, Chen,Liqiong, Zhang,Ying, Lin,Yishuai, Zheng,Xiangkuo, Zhou,Tieli, Chen,Lijiang, Huang,Na, Chen,Tao, Chen,Liqiong, Zhang,Ying, Lin,Yishuai, Zheng,Xiangkuo, Zhou,Tieli, and Chen,Lijiang

Abstract

Na Huang,1 Tao Chen,1 Liqiong Chen,2 Ying Zhang,2 Yishuai Lin,2 Xiangkuo Zheng,1 Tieli Zhou,1 Lijiang Chen1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325035, Zhejiang Province, People’s Republic of China; 2Department of Medical Lab Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, 325035, Zhejiang Province, People’s Republic of ChinaCorrespondence: Lijiang Chen; Tieli ZhouDepartment of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaTel +86-577-8669-9370; +86-577-86689885Email wyychenlijiang@163.com; wyztli@163.comPurpose: This study aimed to evaluate the in vitro activity of meropenem-vaborbactam (MVB) against a collection of carbapenem-resistant Escherichia coli (CREC) isolates and to compare the activity with other antibiotics with regard to different separation sites, carbapenem-resistant mechanisms, and sequence types (STs).Methods: A total of 58 CREC strains were used as the experimental strains from the First Affiliated Hospital of Wenzhou Medical University in southeastern China. The minimum inhibitory concentrations of MVB, ceftazidime-avibactam, and tigecycline against all the experimental strains were determined by the microdilution broth method.Results: MVB exhibited higher antimicrobial activity (83% susceptibility) than that of other antibiotics, except for colistin and tigecycline. The susceptibility of CREC strains towards MVB varied with regard to carbapenem-resistant mechanisms and STs, especially in Klebsiella pneumoniae carbapenemase (KPC)-positive isolates and ST8 isolates.Conclusion: MVB exhibited considerably high activity against KPC-producing and ST8 CREC isolates. It has the great potential to be an alternative for the treatment of infections caused by CREC after determining the type of carbapenemase, the susceptibility to MVB and/or STs.Keyw

Published
2021

141. Synergistic Activity and Biofilm Formation Effect of Colistin Combined with PFK-158 Against Colistin-Resistant Gram-Negative Bacteria

Author

Chen,Liqiong, Yu,Kaihang, Chen,Lijiang, Zheng,Xiangkuo, Huang,Na, Lin,Yishuai, Jia,Huaiyu, Liao,Wenli, Cao,Jianming, Zhou,Tieli, Chen,Liqiong, Yu,Kaihang, Chen,Lijiang, Zheng,Xiangkuo, Huang,Na, Lin,Yishuai, Jia,Huaiyu, Liao,Wenli, Cao,Jianming, and Zhou,Tieli

Abstract

Liqiong Chen,1 Kaihang Yu,2 Lijiang Chen,1 Xiangkuo Zheng,1 Na Huang,1 Yishuai Lin,2 Huaiyu Jia,1 Wenli Liao,1 Jianming Cao,2 Tieli Zhou1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 2Department of Medical Laboratory Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaCorrespondence: Tieli Zhou Tel +86-577-86689885Email wyztli@163.comJianming Cao Tel +86-577-88069595Email wzcjming@163.comPurpose: The emergence of colistin resistance among Gram-negative bacteria (GNB) poses a serious public health threat. Therefore, it is necessary to enhance the antibacterial activity of colistin through the combination with other drugs. In this study, we demonstrated the synergistic activity and the possible synergy mechanism of colistin with PFK-158 against colistin-resistant GNB, including non-fermenting bacteria and Enterobacteriaceae.Patients and Methods: Thirty-one colistin-resistant GNB, including Pseudomonas aeruginosa (n = 9), Acinetobacter baumannii (n = 5), Escherichia coli (n = 8) and Klebsiella pneumoniae (n = 9), were collected as the experimental strains and the minimum inhibitory concentrations (MICs) of colistin, other routine antimicrobial agents and PFK-158 against all strains were determined by the broth microdilution method. The synergistic activity of colistin with PFK-158 was assessed by the checkerboard assay and time-kill assay. The biofilm formation assay and scanning electron microscopy were used to demonstrate the biofilm formation effect of colistin with PFK-158 against colistin-resistant GNB.Results: The results of the checkerboard assay showed that when colistin was used in combination with PFK-158, synergistic activity was observed against the 31 colistin-resistant GNB. The time-kill assay presented a significant killing activity of colisti

Published
2021

142. Unraveling Mechanisms and Epidemic Characteristics of Nitrofurantoin Resistance in Uropathogenic Enterococcus faecium Clinical Isolates

Author

Zhang,Ying, Wang,Lingbo, Zhou,Cui, Lin,Yishuai, Liu,Shixing, Zeng,Weiliang, Yu,Kaihang, Zhou,Tieli, Cao,Jianming, Zhang,Ying, Wang,Lingbo, Zhou,Cui, Lin,Yishuai, Liu,Shixing, Zeng,Weiliang, Yu,Kaihang, Zhou,Tieli, and Cao,Jianming

Abstract

Ying Zhang,1 Lingbo Wang,2 Cui Zhou,2 Yishuai Lin,1 Shixing Liu,2 Weiliang Zeng,1 Kaihang Yu,1 Tieli Zhou,2 Jianming Cao1 1Department of Medical Lab Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, 325035, People’s Republic of China; 2Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, 325035, People’s Republic of ChinaCorrespondence: Jianming CaoDepartment of Medical Lab Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaTel +86-0577-88069595Email wzcjming@163.comTieli ZhouDepartment of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of ChinaTel +86-0577-86689885Email wyztli@163.comPurpose: Multidrug-resistant (MDR) Enterococcus faecium is an important nosocomial pathogen causing urinary tract infection, and the reapplication of nitrofurantoin (NIT) in the clinic has attracted great attention. This study aims to explore the NIT resistance mechanisms and epidemiological characteristics of E. faecium clinical isolates.Patients and Methods: A total of 633 E. faecium clinical isolates was obtained from urine samples in a clinical teaching hospital during 2017– 2018. Among them, 40 NIT-resistant strains, and a similar number of -intermediate and -susceptible strains were isolated. The minimum inhibitory concentrations (MICs) of NIT were detected by agar dilution method. The prevalence and mutations of nitroreductase-encoding genes ef0404 and ef0648 were explored by polymerase chain reaction (PCR), followed by efflux pump inhibition test and quantitative real-time PCR (qRT-PCR) to investigate the resistance mechanisms of NIT. Furthermore, the epidemiological characteristics were detected by multilocus sequence typing (MLST)

Published
2021

143. Resistance Profiles and Biological Characteristics of Rifampicin-Resistant Staphylococcus aureus Small-Colony Variants

Author

Zheng,Xiangkuo, Fang,Renchi, Wang,Chong, Tian,Xuebin, Lin,Jie, Zeng,Weiliang, Zhou,Tieli, Xu,Chunquan, Zheng,Xiangkuo, Fang,Renchi, Wang,Chong, Tian,Xuebin, Lin,Jie, Zeng,Weiliang, Zhou,Tieli, and Xu,Chunquan

Abstract

Xiangkuo Zheng,1,* Renchi Fang,2,* Chong Wang,3 Xuebin Tian,4 Jie Lin,1 Weiliang Zeng,4 Tieli Zhou,1 Chunquan Xu1 1Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325035, People’s Republic of China; 2Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310003, People’s Republic of China; 3Department of Laboratory Medicine, Qingdao Municipal Hospital, Qingdao, 266000, People’s Republic of China; 4School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, 325035, People’s Republic of China*These authors contributed equally to this workCorrespondence: Chunquan Xu; Tieli ZhouDepartment of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, People’s Republic of ChinaTel +86-577-5557-9725; +86-577-8668-9885Email 417242374@qq.com; wyztli@163.comBackground: Staphylococcus aureus (S. aureus) is a major contributor to nosocomial and community-acquired infections. S. aureus small colony variants (SCVs) which changed in relevant phenotype have made more limited and difficult for therapeutic options against S. aureus infections increasingly. Rifampicin is considered as the “last-resort” antibiotic against S. aureus. Our study investigated resistance profiles and biological characteristics of rifampicin-resistant S. aureus SCVs.Methods: We collected S. aureus SCVs that were selected from 41 rifampicin-resistant clinical isolates. Then, biological characteristics, resistance spectrum, and rifampicin resistance mechanisms of tested S. aureus SCVs and corresponding parental strains were investigated by classic microbiological methods, agar dilution method, polymerase chain reaction (PCR). Moreover, the fitness cost of S. aureus SCVs, including growth, biofilm formation ability, and virulence profile, was also determined b

Published
2021

144. Invasive candidiasis in intensive care units in China: in vitro antifungal susceptibility in the China-SCAN study

Author

Liu, Wei, Tan, Jingwen, Sun, Jimei, Xu, Zhijiang, Li, Min, Yang, Qing, Shao, Haifeng, Zhang, Liyan, Liu, Weixia, Wan, Zhe, Cui, Wei, Zang, Bin, Jiang, Dongpo, Fang, Qiang, Qin, Bingyu, Qin, Tiehe, Li, Weiqin, Guo, Fengmei, Liu, Dawei, Guan, Xiandong, Yu, Kaijiang, Qiu, Haibo, Li, Ruoyu, Jin, Jun, Zhan, Ying, Qian, Xuefeng, Liu, Lijun, Zhu, Xueming, Wang, Difen, Liang, Xianquan, Fei, Ying, Lu, Huixia, Ma, Bing, Zhu, Duming, Hu, Bijie, Lin, Jiandong, Yang, Bin, Li, Maoqin, Ding, Chen, Qin, Yingzhi, Feng, Yongwang, Zhao, Hongsheng, Zhu, Shaopeng, Cao, Quan, Zhou, Suming, Mei, Yaning, Zhao, Xiaoqin, Tang, Zhanhong, Liang, Hongjie, Zhu, Jianhuan, Shi, Yingjiao, Xu, Yingchun, Liu, Jian, Yao, Liqiong, Yang, Xiaojun, Jia, Wei, Zheng, Ruiqiang, Zhu, Xiaoping, Xu, Yuan, Lu, Xinxin, Zhao, Mingyan, Lu, Juan, Guan, Xiangdong, Liao, Kang, Wu, Dawei, Sun, Enhua, Lin, Xifang, Zhou, Tieli, Xing, Jinyan, Liu, Pengpeng, Chen, Dechang, Wang, Hu, Wang, Xue, Shi, Qindong, Ai, Yuhang, Zou, Mingxiang, Zhou, Jianxin, Tang, Mingzhong, Song, Qing, Shen, Dingxia, Guo, Weiyuan, Li, Jianguo, Guo, Qinglian, Qian, Chuanyun, Feng, Xia, Wan, Xianyao, Zhang, Jiuzhi, Hu, Zhenjie, Zhang, Jinyan, Wang, Yongqiang, Zhang, Jianlei, Yan, Jie, Zhou, Lizhen, Yu, Xiangyou, Ji, Ping, Xiong, Jianqiong, Zhang, Lei, Yin, Yongjie, Liu, Haibo, Yuan, Shiying, Zeng, Ji, Li, Shusheng, Li, Li, Yan, Jing, Xiao, Zhen, Hospital, Zhejiang, Wang, Chunting, Wang, Yong, Sun, Rongqing, Xu, Min, and Wan, Zhe

Published
2014
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146. Characteristics of rare ST463 carbapenem-resistant Pseudomonas aeruginosaclinical isolates from blood

Author

Zhang, Xiaodong, Tang, Miran, Xu, Ye, Xu, Mengxin, Qian, Changrui, Zheng, Xiangkuo, Zhou, Tieli, and Wu, Qing

Abstract

•The blaKPC-2gene caused carbapenem resistance to TL3773.•GyrA and ParC mutations may confer fluoroquinolone resistance to TL3773.•ST463 Pseudomonas aeruginosawas highly related to blaKPC-2and exoU.•High pyocyanin and biofilm formation of TL3773 may cause patients’ poor•prognosis.•ST463 P. aeruginosashowed a rapid transmission phenomenon.

Published
2023
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