Characterization of Fosfomycin Resistant Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Human and Pig in Taiwan

Sung-Pin Tseng; Sheng-Fan Wang; Cheng-Yu Kuo; Jun-Wei Huang; Wei-Chun Hung; Guan-Ming Ke; Po-Liang Lu

doi:10.1371/journal.pone.0135864

Characterization of Fosfomycin Resistant Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Human and Pig in Taiwan

PLoS One. 2015 Aug 17;10(8):e0135864. doi: 10.1371/journal.pone.0135864. eCollection 2015.

Authors

Sung-Pin Tseng¹, Sheng-Fan Wang², Cheng-Yu Kuo³, Jun-Wei Huang², Wei-Chun Hung⁴, Guan-Ming Ke⁵, Po-Liang Lu⁶

Affiliations

¹ Department of Medical Laboratory Science and Biotechnology, College of Health Sciences, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC; Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwan, ROC.
² Department of Medical Laboratory Science and Biotechnology, College of Health Sciences, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC.
³ Division of Infectious Diseases, Kaohsiung Medical University Chung-Ho Memorial hospital, Kaohsiung, Taiwan, ROC; Department of Internal Medicine, Ministry of Health and Welfare Pingtung Hospital, Kaohsiung, Taiwan, ROC.
⁴ Department of Microbiology and Immunology, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC.
⁵ Graduate Institute of Animal Vaccine Technology, National Pingtung University of Science and Technology, Neipu, Pingtung, Taiwan, ROC.
⁶ Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, ROC; College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC; Department of Internal Medicine, Kaohsiung Medical University Hospital, Taiwan, Kaohsiung, ROC.

Abstract

To investigate the efficacy of fosfomycin against extended-spectrum β-lactamases (ESBL) producing Escherichia coli in Taiwan and the resistance mechanisms and characterization of human and pig isolates, we analyzed 145 ESBL-producing isolates collected from two hospitals (n = 123) and five farms (n = 22) in Taiwan from February to May, 2013. Antimicrobial susceptibilities were determined. Clonal relatedness was determined by PFGE and multi-locus sequence typing. ESBLs, ampC, and fosfomycin resistant genes were detected by PCR, and their flanking regions were determined by PCR mapping and sequencing. The fosfomycin resistant mechanisms, including modification of the antibiotic target (MurA), functionless transporters (GlpT and UhpT) and their regulating genes such as uhpA, cyaA, and ptsI, and antibiotic inactivation by enzymes (FosA and FosC), were examined. The size and replicon type of plasmids carrying fosfomycin resistant genes were analyzed. Our results revealed the susceptibility rates of fosfomycin were 94% for human ESBL-producing E. coli isolates and 77% for pig isolates. The PFGE analysis revealed 79 pulsotypes. No pulsotype was found existing in both human and pig isolates. Three pulsotypes were distributed among isolates from two hospitals. ISEcp1 carrying blaCTX-M-group 9 was the predominant transposable elements of the ESBL genes. Among the thirteen fosfomycin resistant isolates, functionless transporters were identified in 9 isolates. Three isolates contained novel amino acid substitutions (Asn67Ile, Phe151Ser and Trp164Ser, Val146Ala and His159Tyr, respectively) in MurA (the target of fosfomycin). Four isolates had fosfomycin modified enzyme (fosA3) in their plasmids. The fosA3 gene was harboured in an IncN-type plasmid (101 kbp) in the three pig isolates and an IncB/O-type plasmid (113 kbp) in the human isolate. In conclusion, we identified that 6% and 23% of the ESBL-producing E. coli from human and pigs were resistant to fosfomycin, respectively, in Taiwan. No clonal spread was found between human and pig isolates. Functionless transporters were the major cause of fosfomycin resistance, and the fosA3-transferring plasmid between isolates warrants further monitoring.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Anti-Bacterial Agents / pharmacology
DNA Transposable Elements / genetics
DNA, Bacterial / genetics
Drug Resistance, Bacterial / genetics*
Escherichia coli / drug effects*
Escherichia coli / genetics*
Escherichia coli Infections / drug therapy
Escherichia coli Infections / microbiology
Escherichia coli Infections / veterinary
Fosfomycin / pharmacology*
Humans
Microbial Sensitivity Tests / methods
Plasmids / genetics
Swine
Taiwan
beta-Lactamases / genetics*

Substances

Anti-Bacterial Agents
DNA Transposable Elements
DNA, Bacterial
Fosfomycin
beta-Lactamases

Grants and funding

This work was supported by Ministry of Science and Technology (MOST 103-2320-B-037-023) and NSYSU-KMU Joint Research Project (NSYSUKMU103-I005) and Kaohsiung Medical University Hospital, Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.