Comparison of antibiotic resistance and virulence in vancomycin-susceptible and vancomycin-resistant Enterococcus faecium strains

Anna Sieńko; Sławomir Czaban; Dominika Ojdana; Piotr Majewski; Anna Wieczorek; Paweł Sacha; Elżbieta Anna Tryniszewska; Piotr Wieczorek

doi:10.20883/jms.288

Authors

Anna Sieńko Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Sławomir Czaban Department of Anaesthesiology and Intensive Therapy, Faculty of Health Science, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Dominika Ojdana Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Piotr Majewski Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Anna Wieczorek Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Paweł Sacha Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Elżbieta Anna Tryniszewska Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland
Piotr Wieczorek Department of Microbiological Diagnostics and Infectious Immunology, Faculty of Pharmacy, Medical University of Bialystok, 15a Waszyngtona Street, 15-269 Bialystok, Poland

DOI:

https://doi.org/10.20883/jms.288

Keywords:

Enterococcus faecium, VRE, VSE, resistance, virulence

Abstract

Aim. Today, infections caused by vancomycin-resistant Enterococcus faecium (VRE) are a major problem in the healthcare system. The aim of this study was to compare the antibiotic resistance and virulence traits between vancomycin-susceptible E. faecium (VSE) and VRE clinical isolates.
Material and Methods. Studies were performed on 66 E. faecium (32 VRE and 34 VSE) strains. Susceptibility testing and identification were performed, and strains were examined for ß-lactamase, hemolysin and biofilm production. Isolates were tested for the presence of 5 van genes, 8 virulence genes and 6 aminoglycoside-modifying enzyme (AME) genes. Obtained amplicons were subjected to electrophoretical separation and DNA sequencing.
Results. Among 32 VRE isolates, 28 were found to have the VanA phenotype, and 4 the VanB. The most frequent resistance and virulence profile among VRE strains was resistance to ampicillin, imipenem, gentamicin, streptomycin, teicoplanin, and vancomycin with enterococcal surface protein (esp), endocarditis antigen (efaA), collagen adhesin (acm), and hialuronidase (hyl) genes; among VSE: resistance to ampicillin, imipenem, gentamicin, streptomycin with esp, efaA, acm, and hyl genes.
Conclusions. Our findings prove that both VRE and VSE strains were well equipped with virulence and resistance genes, although VRE strains were characterized by a greater variety and a higher number of these genes. However, statistical analysis revealed no significant differences between VSE and VRE strains (p > 0.05). Nevertheless, our results suggest that VRE strains may slowly acquire and incorporate resistance and virulence genes, due to their ability to survive in a hospital environment for a long time.

Downloads

Download data is not yet available.

References

Cheng V, Chen J, Tai J, Wong S, Poon R, Hung I, et al. Decolonization of gastrointestinal carriage of vancomycin‑resistant Enterococcus faecium: case series and review of literature. BMC Infect Dis. 2014;14:514.

Amyes SG. Enterococci and streptococci. Int J Antimicrob Agents. 2007;29(3):43–52.

Buultjens AH, Lam MC, Ballard S, Monk IR, Mahony AA, Grabsch EA, et al. Evolutionary origins of emergent ST796 clone of vancomycin resistant Enterococcus faecium. PeerJ. 2017;5:e2916.

Sava IG, Heikens E, Huebner J. Pathogenesis and immunity in enterococcal infections. Clin Microbiol Infect. 2010;16(6):533–540.

Raven KE, Reuter S, Reynolds R, Broderick HJ, Russell JE, Estee Torok M, et al. A decade of genomic history for healthcare‑associated Enterococcus faecium in the United Kingdom and Ireland. Genome Res. 2016;26(10):1388–1396.

Comerlato CB, Resende MC, Caierão J, d'Azevedo PA. Presence of virulence factors in Enterococcus faecalis and Enterococcus faecium susceptible and resistant to vancomycin. Mem Inst Oswaldo Cruz. 2013;108(5):590–595.

Diarmaid H. Exploiting genomics, genetics and chemistry to combat antibiotic resistance. Nat Rev Gen. 2003;4(6):432–441.

Arias CA, Murray BE. The rise of the Enterococccus: beyond vancomycin resistance. Nat Rev Microbiol. 2012;10:266–278.

Lebreton F, Depardieu F, Bourdon N, Fines‑Guyon M, Berger P, Camiade S, et al. D‑Ala‑d-Ser VanN‑type transferable vancomycin resistance in Enterococcus faecium. Antimicrob Agents Chemother. 2011;55(10):4606–4012.

Xu X, Lin D, Yan G, Ye X, Wu S, Guo Y, et al. vanM, a new glycopeptides resistance gene cluster found in Enterococcus faecium. Antimicrob. Agents. Chemother. 2010;52(7):2667–2672.

Cesar A, Contreas MD, German A. Clinical Aspects of Multidrug Resistant Enterococci. Antibiotic Discovery and Development, Springer, US, 2012;617–648.

Ramirez MS, Tolmasky ME. Aminoglycoside modifying enzymes. Drug Resist Update. 2010;13:151–171.

Kowalska‑Krochmal B, Dworniczek E, Dolna I, Bania J, Wałecka E, Seniuk A, et al. Resistance patterns and occurrence of virulence determinants among GRE strains in southwestern Poland. Adv Med Sci. 2011;56(2):304–310.

Lall N, Basak S. High level aminoglycoside resistant Enterococcus species: a study. Int J Curr Res. 2014;6(3):16–21.

Di Rosa R, Creti R, Venditti M, D'Amelio R, Arciola CR, Montanaro L, et al. Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium. FEMS Microbiol Lett. 2006;256(1):145–150.

Fisher K, Phillips C. The ecology, epidemiology and virulence of Enterococcus. Microbiology. 2009;155:1749–1757.

Sieńko A, Wieczorek P, Majewski P, Ojdana D, Wieczorek A, Olszańska D, et al. Comparison of antibiotic resistance and virulence between biofilm‑producing and non‑producing clinical isolates of Enterococcus faecium. Acta Biochim Pol. 2015;4(62):859–866.

Özden Tuncer B, Ay Z, Tuncer Y. Occurrence of enterocin genes, virulence factors, and antibiotic resistance in 3 bacteriocin‑producer Enterococcus faecium strains isolated from Turkish tulum cheese. Turk J Biol. 2013;37:443–449.

Gałkowska H, Olszewski WL, Podbielska A. Staphylococcal and enterococcal virulence – a review. Centr Eur J Immunol. 2011;36(1):56–64.

Singh KV, Nallaparedy SR, Sillanpaa J, Murray BE. Importance of the collagen adhesin Ace in pathogenesis and protection against Enterococcus faecalis experimental endocarditis. PLoS Pathog. 2010;8:e1000716.

Praharaj I, Sujatha S, Parija SC. Phenotypic & genotypic characterization of vancomycin resistant Enterococcus isolates from clinical specimens. Indian J Med Res. 2013;138(4):549–556.

Mohamed JA, Huang DB. Biofilm formation by enterococci. J Med Microbiol. 2007;56(12):1581–1588.

Camargo ILBC, Gilmore MS, Darini ALC. Multilocus sequence typing and analysis of putative virulence factors in vancomycin‑resistant and vancomycin‑sensitive Enterococcus faecium isolates from Brazil. Clin Microbiol Infect. 2006;12:1123–1130.

Iris N, Sayıner HS, Yıldırmak T, Şimşek F. Arat ME. Distribution of vancomycin resistant enterococci and their resistance patterns determined by surveillance. Afr J Microbiol Res. 2014;8(7):680–684.

Simonsen GS, Småbrekke L, Monnet DL, Sørensen TL, Møller JK, Kristinsson KG, et al. Prevalence of resistance to ampicillin, gentamicin and vancomycin in Enterococcus faecalis and Enterococcus faecium isolates from clinical specimens and use of antimicrobials in five Nordic hospitals. J Antimicrob Chemother. 2003;51(2):323–331.

Yazgi H, Ertek M, Erol E, Ayyldiz A. A Comparison of High‑level Aminoglycoside Resistance in Vancomycin‑sensitive and Vancomycin‑reisistant Enterococcus species. J Int Med Res. 2002;30:529–534.

Alotaibi FE, Bukhari EE. Emergence of Vancomycin‑resistant Enterococci at a Teaching Hospital, Saudi Arabia. Chin Med J. 2017;130:340–346.

Fernandes SC, Dhanashree B. Drug resistance and virulence determinants in clinical isolates of Enterococcus species. Indian J Med Res. 2013;137(5):981–985.

Manavalan J, Kannaiyan K, Velayutham A, Vadivel S, Kuthalaramalingam S. Phenotypic speciation of enterococci with special reference to prevalence, virulence and antimicrobial resistance. Int J Res Med Sci. 2015;3(10):2623–2629.

Dutka‑Mahlen S, Evers S, Courvalin P. Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR. J Clin Microbiol. 1995;33:24–27.

Pitkälä A, Salmikivi L, Bredbacka P, Myllyniemi AL, Koskinen MT. Comparison of tests for detection of beta‑lactamase‑producing staphylococci. J Clin Microbiol. 2007;45:2031–2033.

Vergis EN, Shankar N, Chow JW, Hayden MK, Snydman DR, Zervos MJ, et al. Association between the presence of enterococcal virulence factors gelatinase, hemolysin, and enterococcal surface protein and mortality among patients with bacteremia due to Enterococcus faecalis. Clin Infect Dis. 2002;35:570–575.

Cabrera‑Contreras R, Morelos‑Ramirez R, Galicia‑Camacho AN, Melendez‑Herrada E. Antibiotic resistance biofilm production in Staphylococcus epidermidis strains, isolated from a Tertiary Care Hospital in Mexico City. ISRN Microbiol. 2013;1–5.

Oliveira A, Cunha MDL. Comparison of methods for the detection of biofilm production in coagulase‑negative staphylococci. BMC Res Notes. 2010;3:260.

Courvalin P, Depardieu F, Perichon B. Detection of the van Alphabet and Identification of Enterococci and Staphylococci at the Species Level by Multiplex PCR. J Clin Microbiol. 2004;42(12):5857–5860.

Zou LK, Wang HN, Zeng B, Li JN, Li XT, Zhang AY, et al. Erythromycin resistance and virulence genes in Enterococcus faecalis from swine in China. New Microbiol. 2011;34:73–80.

Padmasini E, Padmaraj R, Sri Vani Ramesh S. High Level Aminoglycoside Resistance and Distribution of Aminoglycoside Resistant Genes among Clinical Isolates of Enterococcus Species in Chennai, India. Sci World J. 2014;329157.

Vakulenko SB, Donabedian SM, Voskresenskiy AM, Zervos MJ, Lerner SA, Chow JW. Multiplex PCR for Detection of Aminoglycoside Resistance Genes in Enterococci. Antimicrob Agents Chemother. 2003;47(4):1423–1426.

Baldir G, Engin DO, Kucukercan M, Inan A, Akcay S, Ozyuker S, et al. High‑level resistance to aminoglycoside, vancomycin and linezolid in enterococci strains. J Microbiol Infect Dis. 2013;3(3):100–103.

Tripathi A, Shukla SK, Singh A, Prasad KN. Prevalence, outcome and risk factor associated with vancomycin‑resistant Enterococcus faecalis and Enterococcus faecium at a Tertiary Care Hospital in Northern India. Indian J Med Microbiol. 2016;34:38–45.

Hasani A, Sharifi Y, Ghotaslou R, Naghili B, Hasani A, Aghazadeh M, et al. Molecular screening of virulence genes in high level gentamicin resistant Enterococcus faecalis and Enterococcus faecium isolated from clinical specimens in northwest Iran Indian J Med Microbiol. 2012;30(2):175–181.

Helmi H, AboulFadl L, El‑Dine SS, El‑Defrawy I. Molecular characterization of Antibiotic Resistant Enterococci. Int J Inf Dis. 2008;3(1):67–75.

Khani M, Fattollahzade M, Pajavand H, Bakhtiari S, Abiti R. Increasing prevalence of Aminoglicoside‑Resistant Enterococcus faecalis Isolates Due to the aac(6’)-aph(2’’) Gene: A Therapeutic Problem in Kermanshah, Iran. Jundishapur J Microbiol. 2016;9(3):e28923.

Leavis HL, Bonten MJ, Willems RJ. Identification of high‑risk enterococcal clonal complexes: Global dispersion of antibiotic resistance. Curr Opin Microbiol. 2006;9:454–460.

Sivertsen A, Billstrom H, Melefors O, Liljequist BO, Wisell KT, Ullberg M, et al. A Multicentre Hospital Outbreak in Sweden Caused by Introduction of a vanB2 Transposon into a Stably Maintained pRUM‑Plasmid in an Enterococcus faecium ST192 Clone. PLoS ONE. 2014;9:e103274.

Vankerckhoven V, Van Autgaerden T, Vael C, Lammens C, Chapelle S, Rossi R, et al. Development of a Multiplex PCR for the Detection of asa1, gelE, cylA, esp, and hyl Genes in Enterococci and Survey for Virulence Determinants among European Hospital Isolates of Enterococcus faecium. J Clin. Microbiol. 2004;42(10):4473–4479.

Saba Copur S, Sahin F, Gocmen JS. Determination of virulence and multidrug resistance genes with polymerase chain reaction method in vancomycin‑sensitive and –resistant enterococci isolated from clinical samples. Turk J Med Sci. 2016;46:877–891.

Biswas PP, Dey S, Sen A, Adhikari L. Molecular Characterization of Virulence Genes in Vancomycin‑Resistant and Vancomycin‑Sensitive Enterococci. J Glob Infect Dis. 2016;8(1):16–24.

Bonten MJ, Willems R, Weinstein RA. Vancomycin‑resistant enterococci: why are they here, and where do they come from? Lancet Infect Dis. 2001;1(5):241–248.

Kawalec M, Gniadkowski M, Hryniewicz W. Outbreak of vancomycin‑resistant enterococci in a hospital in Gdańsk, Poland. J Clin Microbiol. 2000;38(9):3317–3322.