Virus Genes

, Volume 54, Issue 6, pp 804–811 | Cite as

Characterization and genome analysis of novel phage vB_EfaP_IME195 infecting Enterococcus faecalis

  • Ronghuan Wang
  • Shaozhen Xing
  • Feiyang Zhao
  • Ping Li
  • Zhiqiang Mi
  • Taoxing Shi
  • Hui LiuEmail author
  • Yigang TongEmail author


Enterococcus faecalis is one of the main bacteria in the human and animal intestine but is also classed as an opportunistic pathogen. During normal growth, E. faecalis produces natural antibiotics and is conducive to human health. As ectopic parasites, E. faecalis is capable of causing infective endocarditis, neonatal sepsis, bloodstream infections, bacteremia, and intraabdominal infections. With the incidence of antibiotic resistance reaching crisis point, it is imperative to find alternative treatments for multidrug-resistant infections. Using phage for pathogen control is a promising treatment option to combat bacterial resistance. In this study, a lytic phage, designated vB_EfaP_IME195, was isolated from hospital sewage using a clinical multidrug-resistant Enterococcus faecalis strain as an indicator. The one-step growth curve with the optimal multiplicity of infection of (MOI) 0.01 revealed a latent period of ~ 30 min and a burst size of ~ 120 plaque-forming units (pfu) per cell. Transmission electron microscopy showed that the phage belongs to the family Podoviridae. Phage vB_EfaP_IME195 has a linear, double-stranded DNA genome of 18,607 bp with a G + C content of 33% and 27 coding sequences (GenBank accession no. KT932700). Run-off sequencing experiments showed that the phage has a unique 59-bp inverted repeat sequences at the terminal ends. BLASTn analysis revealed that vB_EfaP_IME195 shares 92% identity (93% genome coverage) with unpublished E. faecalis phage Idefix. This study reported a novel E. faecalis phage with unique genome termini containing inverted repeats. The isolation and characterization of this novel lytic E. faecalis phage provides the basis for the development of new therapeutic agents like phage cocktails for multidrug-resistant E. faecalis infection, and its unique genomic feature would also provide valuable knowledge and insight for further phage genome analysis.


Enterococcus faecalis Phage Biological characteristics Genome Termini 


Author Contributions

Yigang Tong, Hui Liu, Taoxing Shi, and Zhiqiang Mi conceived and designed the experiments and critically evaluated the manuscript. Ronghuan Wang was responsible for data and sequence analyses and wrote the manuscript. Shaozhen Xing isolated and identified the phage and conducted the biological characterization experiments. Feiyang Zhao conducted the sequencing experiments. Ping LI extracted the phage nucleotide. All authors read and approved the final manuscript.


This research was supported by a grant from The National Key Research and Development Program of China (2015AA020108, 2016YFC1202705, AWS16J020, and AWS15J006), the National Natural Science Foundation of China (81572045, 81672001, and 81621005), and the State Key Laboratory of Pathogen and Biosecurity (SKLPBS1518).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. 1.
    Jett BD, Huycke MM, Gilmore MS (1994) Virulence of enterococci. Clin Microbiol Rev 7:462–478CrossRefGoogle Scholar
  2. 2.
    Maki DG, Agger WA (1988) Enterococcal bacteremia: clinical features, the risk of endocarditis, and management. Medicine 67:248–269CrossRefGoogle Scholar
  3. 3.
    Richards MJ, Edwards JR, Culver DH, Gaynes RP (2000) Nosocomial infections in combined medical-surgical intensive care units in the United States. Infect Control Hosp Epidemiol 21:510–515CrossRefGoogle Scholar
  4. 4.
    Saito HE, Harp JR, Fozo EM (2017) Enterococcus faecalis responds to individual exogenous fatty acids independently of saturation or chain length. Appl Environ Microbiol 84:e01633–e01617PubMedPubMedCentralGoogle Scholar
  5. 5.
    Karki S, Houston L, Land G, Bass P, Kehoe R, Borrell S, Watson K, Spelman D, Kennon J, Harrington G (2012) Prevalence and risk factors for VRE colonisation in a tertiary hospital in Melbourne, Australia: a cross sectional study. Antimicrob Resistance Infect Control 1(1):31–31CrossRefGoogle Scholar
  6. 6.
    Klare I, Witte W, Wendt C, Werner G (2012) Vancomycin-resistant enterococci (VRE). Recent results and trends in development of antibiotic resistance. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz 55:1387CrossRefGoogle Scholar
  7. 7.
    Organization WH (2014) Antimicrobial resistance: global report on surveillance. Australas Med J 7:237Google Scholar
  8. 8.
    Nilsson AS (2014) Phage therapy–constraints and possibilities. Ups J Med Sci 119:192–198CrossRefGoogle Scholar
  9. 9.
    Drulis-Kawa Z, Majkowska-Skrobek G, Maciejewska B (2015) Bacteriophages and phage-derived proteins–application approaches. Curr Med Chem 22(14):1757–1773CrossRefGoogle Scholar
  10. 10.
    Henry M, Lavigne R, Debarbieux L (2013) Predicting in vivo efficacy of therapeutic bacteriophages used to treat pulmonary infections. Antimicrob Agents Chemother 57:5961–5968CrossRefGoogle Scholar
  11. 11.
    Tsonos J, Oosterik LH, Tuntufye HN, Klumpp J, Butaye P, De GH, Hernalsteens JP, Lavigne R, Goddeeris BM (2014) A cocktail of in vitro efficient phages is not a guarantee for in vivo therapeutic results against avian colibacillosis. Vet Microbiol 171:470–479CrossRefGoogle Scholar
  12. 12.
    Sulakvelidze A, Alavidze Z, Morris JJ (1934) Bacteriophage therapy. Antimicrob Agents Chemother 2:1110–1110Google Scholar
  13. 13.
    Flaherty JE, Harbaugh BK, Jones JB, Somodi GC, Jackson LE (2001) H-mutant bacteriophages as a potential biocontrol of bacterial blight of geranium. Hortscience 36:98–100Google Scholar
  14. 14.
    Guenther S, Huwyler D, Richard S, Loessner MJ (2009) Virulent bacteriophage for efficient biocontrol of listeria monocytogenes in ready-to-eat foods. Appl Environ Microbiol 75:93–100CrossRefGoogle Scholar
  15. 15.
    Sulakvelidze A (2013) Using lytic bacteriophages to eliminate or significantly reduce contamination of food by foodborne bacterial pathogens. J Sci Food Agric 93:3137–3146CrossRefGoogle Scholar
  16. 16.
    Chen Y, Sun E, Song J, Yang L, Wu B (2018) Complete genome sequence of a novel T7-like bacteriophage from a Pasteurella multocida capsular type A isolate. Curr Microbiol 75:574–579CrossRefGoogle Scholar
  17. 17.
    Ellis EL, Max D (1939) The growth of bacteriophage. J Gen Physiol 22:365–384CrossRefGoogle Scholar
  18. 18.
    Xing S, Zhang X, Qiang S, Jian W, Mi Z, Pei G, Yong H, An X, Fu K, Zhou L (2017) Complete genome sequence of a novel, virulent Ahjdlikevirus bacteriophage that infects Enterococcus faecium. Arch Virol 162:1–5CrossRefGoogle Scholar
  19. 19.
    Gan HM, Sieo CC, Tang SGH, Omar AR, Yin WH (2013) The complete genome sequence of EC1-UPM, a novel N4-like bacteriophage that infects Escherichia coli O78:K80. Virol J 10:308–308CrossRefGoogle Scholar
  20. 20.
    Li P, Chen B, Song Z, Song Y, Yang Y, Ma P, Wang H, Ying J, Ren P, Yang L (2012) Bioinformatic analysis of the Acinetobacter baumannii phage AB1 genome. Gene 507:125–134CrossRefGoogle Scholar
  21. 21.
    Yang X, Wang Q, Liang B, Wu F, Li H, Liu H, Sheng C, Ma Q, Yang C, Xie J (2017) An outbreak of acute respiratory disease caused by a virus associated RNA II gene mutation strain of human adenovirus 7 in China, 2015. PLos ONE 12:e0172519CrossRefGoogle Scholar
  22. 22.
    Jamal M, Hussain T, Das CR, Andleeb S (2015) Isolation and characterization of a Myoviridae MJ1 bacteriophage against multi-drug resistant Escherichia coli 3. Jundishapur J Microbiol 8:e25917CrossRefGoogle Scholar
  23. 23.
    Merabishvili M, Vandenheuvel D, Kropinski AM, Mast J, Vos DD, Verbeken G, Noben JP, Lavigne R, Vaneechoutte M, Pirnay JP (2014) Characterization of newly isolated lytic bacteriophages active against Acinetobacter baumannii. PLoS ONE 9(8):e104853CrossRefGoogle Scholar
  24. 24.
    Wu S, Zachary E, Wells K, Loccarrillo C (2013) Phage therapy: future inquiries. Postdoc J 1:24PubMedPubMedCentralGoogle Scholar
  25. 25.
    Ciacci N, D’Andrea MM, Marmo P, Dematte E, Amisano F, Pilato VD, Fraziano M, Lupetti P, Rossolini GM, Thaller MC (2018) Characterization of vB_Kpn_F48, a newly discovered lytic bacteriophage for Klebsiella pneumoniae of sequence type 101. Viruses 10(9):482CrossRefGoogle Scholar
  26. 26.
    Chan BK, Abedon ST, Loccarrillo C (2013) Phage cocktails and the future of phage therapy. Future Microbiol 8:769–783CrossRefGoogle Scholar
  27. 27.
    Gill JJ, Hyman P (2010) Phage choice, isolation, and preparation for phage therapy. Curr Pharm Biotechnol 11(1):2–14CrossRefGoogle Scholar
  28. 28.
    Merabishvili M, Pirnay JP, Verbeken G, Chanishvili N, Tediashvili M, Lashkhi N, Glonti T, Krylov V, Mast J, Parys LV (2009) Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLos ONE 4:e4944CrossRefGoogle Scholar
  29. 29.
    Li X, Ding P, Han C, Fan H, Wang Y, Mi Z, Feng F, Tong Y (2014) Genome analysis of Enterococcus faecalis bacteriophage IME-EF3 harboring a putative metallo-beta-lactamase gene. Virus Genes 49:145–151CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Ronghuan Wang
    • 1
  • Shaozhen Xing
    • 2
  • Feiyang Zhao
    • 3
  • Ping Li
    • 3
  • Zhiqiang Mi
    • 3
  • Taoxing Shi
    • 4
  • Hui Liu
    • 1
    Email author
  • Yigang Tong
    • 3
    • 5
    Email author
  1. 1.School of Public HealthLanzhou UniversityLanzhouChina
  2. 2.Li Ka Shing Faculty of MedicineThe University of Hong KongPokfulamChina
  3. 3.State Key Laboratory of Pathogen and BiosecurityBeijing Institute of Microbiology and EpidemiologyBeijingChina
  4. 4.Academy of Military Medical SciencesBeijingChina
  5. 5.College of Life Science and TechnologyBeijing University of Chemical TechnologyBeijingChina

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