Comparison of biofilm-producing Enterococcus faecalis, Enterococcus faecium, and unusual Enterococcus strains
Keywords:
Enterococcus faecalis, Enterococcus faecium, Biofilm, Resistance, Virulence
Abstract
The present study focused on determining the prevalence of biofilm-forming ability in Enterococ-cus faecalis, E. faecium, and unusual Enterococcus clinical isolates, and comparison of resistance and the prevalence of selected virulence factors among biofilm-positive strains. The ability to form biofilm was detected in 13.3% of E. faecalis, 90% of E. faecium, and 57.1% of unusual Enterococcus strains (p=0.026). All E. faecalis strains were susceptible to β-lactams, while 37.5% of unusual and all E. faecium isolates were resistant to these antibiotics. Resistance to gentamicin was detected in 75% of E. faecalis, 55.5% of E. faecium, and 25% of other strains; resistance to streptomycin in 25%, 83.3%, and 50%, respectively. Analysis of the virulence revealed that the enterococcal surface protein (esp) gene was found in all E. faecium, 75.0% of E. faecalis, and 37.5% of other strains; collagen adhesin gene (ace) in 100%, 25.0%, and 37.5%; and hyaluronidase gene (hyl) in 83.3%, 0%, and 37.5%, respectively. Analysis of the resistance and virulence patterns showed that E. faecium isolates had the greatest variety of virulence and resistance determinants, while the lowest variety was exhibited by unusual strains. These findings indicate that unusual biofilm-producing Enterococcus strains have lower resistance and virulence potency than E. faecalis and E. faecium.
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References
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31. Billström H, Sullivan A, Lund B. Cross-transmission of clinical Enterococcus faecium in relation to esp and antibiotic resistance. J Appl Microbiol. 2008; 105: 2115-2122.
32. Contreras GA, Diaz Granados CA, Cortes L, Reyes J, Vanegas S, et al. Nosocomial outbreak of Enterococcus gallinarum: untaming of rare species of enterococci. J Hosp Infect. 2008; 70: 346-352.
33. Franiczek R, Dolna I, Dworniczek E, Krzyżanowska B, Seniuk A, Piątkowska E. In vitro activity of tigecycline against clinical isolates of Gram-positive and Gram-negative bacteria displaying different resistance phenol-types. Adv Clin Exp Me. 2008; 17: 545-551.
34. Freitas AR, Coque TM, Novais C, Hammerum AM, Lester CH, Zervos MJ, et al. Human and swine hosts share vancomycin-resistant Enterococcus faecium CC17 and CC5 and Enterococcus faecalis CC2 clonal clusters harboring Tn1546 on indistinguishable plasmids. J Clin Microbiol. 2011; 49: 925-931.
35. Sieńko A, Wieczorek P, Wieczorek A, Sacha P, Majewski P, Ojdana D, et al. Occurrence of high-level aminoglycoside resistance (HLAR) among Enterococcus species strains. Prog Health Sci. 2014; 4: 179-187.
36. Werner G, Grforer S, Fleige C, Witte W, Klare I. Tigecycline-resistant Enterococcus faecalis strain isolated from a German Intensive Care Unit Patient. J Antimicrob Chemother. 2008; 61: 1182-1183.
37. 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: 100-103.
38. Dupre I, Zanetti S, Schito AM, Fadda G, Sechi LA. Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected in Sardinia (Italy). J Med Microbiol. 2003; 52: 491-498.
39. Tendolkar PM, Baghdayan AS, Gilmore MS, Shankar N. Enterococcal surface protein, Esp, enhances biofilm formation by Enterococcus faecalis. Infect Imm. 2004; 72: 6032-6039.
40. Hallgren A, Claesson C, Saeedi B, Monstein HJ, Hanberger H, Nilsson LE. Molecular detection of aggregation substance, enterococcal surface protein, and cytolysin genes and in vitro adhesion to urinary catheters of E. faecalis and E. faecium. Int J Med Microbiol. 2009; 299: 323-332.
41. Raad II, Hanna HA, Boktour M, Chaiban G, Hachem RY, Dvorak T, et al. Vancomycin-resistant Enterococcus faecium: catheter colonization, esp gene, and decreased susceptibility to antibiotics in biofilm. Antimicrob Agents Chemother. 2005; 49: 5046-5050.
42. 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: 4473-4479.
43. 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: 590-595.
44. Archimbaud C, Shankar N, Forestier C, Baghdayan A, Gilmore MS, Charbonne F, et al. In vitro adhesive properties and virulence factors of Enterococcus faecalis strains. Res Microbiol. 2002; 153: 75-80.
2. Amyes SG. Enterococci and streptococci. Int J Antimicrob Agents. 2007; 29: 43-52.
3. Marschall J, Piccirillo ML, Fraser VJ, Doherty JA, Warren KW. Catheter removal versus retention in the management of catheter-associated enterococcal bloodstream infections. Can J Infect Dis Med. 2013; 24(3): e83-e87.
4. Fernandes SC, Dhanashree B. Drug resistance and virulence determinants in clinical isolates of Enterococcus species. Indian J Med Res. 2013; 137: 981-985.
5. 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: 145-150.
6. Fisher K, Phillips C. The ecology, epidemiology and virulence of Enterococcus. Microbiology. 2009; 155: 1749-1757.
7. Sava IG, Heikens E, Huebner J. Pathogenesis and immunity in enterococcal infections. Clin Microbiol Infect. 2010; 16: 533-540.
8. Prakash VP, Rao SR, Parija SC. Emergence of unusual species of enterococci causing infections, South India. BMC Infect Dis. 2005; 5: 14.
9. Tan CK, Lai CC, Wang JY, Lin SH, Liao SH, Huang Y, et al. Bacteriemia caused by non-faecalis and non-faecium Enterococcus species at a medical center in Taiwan, 2000 to 2008. J Infect. 2010; 61: 34-43.
10. Kenzaka T, Takamura N, Kumabe A, Takeda K. A case of subacute infective endocarditis and blood access infection caused by Enterococcus durans. BMC Infect Dis. 2013; 13: 594.
11. Heikens E, Bonten MJ, Willems RJ. Enterococcal surface protein Esp is important for biofilm formation of Enterococcus faecium E1162. J Bacteriol. 2007; 189: 8233-8240.
12. Mohamed JA, Huang DB. Biofilm formation by enterococci. J Med Microbiol. 2007; 56: 1581-1588.
13. Paganelli FL, Willems RJ, Jansen P, Hendrickx A, Zhang X, Bonten MJ, et al. Enterococcus faecium biofilm formation: identification of major autolysin AtlAEfm, associated Acm surface localization, and AtlAEfm-independent extracellular DNA Release. MBio. 2013; 4(2): e00154.
14. Almohamad S, Somarajan SR, Singh KV, Nallapareddy SR, Murray BE. Influence of isolate origin and presence of various genes on biofilm formation by Enterococcus faecium. FEMS Microbiol Lett. 2014; 353: 151-156.
15. Kafil HS, Mobarez AM. Assessment of biofilm formation by enterococci isolates from urinary tract infections with different virulence profiles. J King Saud Univ Sci. 2015; 27: 312-317.
16. Iwen PC, Kelly DM, Linder J, Hinrichs SH, Dominguez EA, Rupp ME, et al. Change in prevalence and antibiotic resistance of Enterococcus species isolated from blood cultures over an 8-year period. Antimicrob Agents Chemother. 1997; 41: 494-495.
17. Lleo M, Bonato B, Tafi MC, Caburlotto G, Benedetti D, Canepari P. Adhesion to medical device materials and biofilm formation capability of some species of enterococci in different physiological states. FEMS Microbiol Lett. 2007; 274: 232-237.
18. Christensen GD, Bisno AL, Simpsom WA, Beachey EH. Adherence of slime producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun. 1982; 37: 318-326.
19. Oliveira A, Cunha MD. Comparison of methods for the detection of biofilm production in coagulase-negative staphylococci. BMC Res Notes. 2010; 3: 260.
20. Freeman DJ, Falkiner FR, Keane CT. New method for detecting slime production by coagulase negative staphylococci. J Clin Pathol. 1989; 48: 872-874.
21. 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; 13: 1-5.
22. 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. 2003; 35: 570-575.
23. 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.
24. 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.
25. Tsikrikonis G, Maniatis AN, Labrou M, Ntokou E, Michail G, Daponte A, et al. Differences in biofilm formation and virulence factors between clinical and fecal enterococcal isolates of human and animal origin. Microb Pathog. 2012; 52: 336–343.
26. Baldassarri L, Cecchini R, Bertuccini L, Ammendolia MG, Iosi F, Arciola CR, et al. Enterococcus spp. produces slime and survives in rat peritoneal macrophages. Med Microbiol Immunol. 2001; 190: 113-120.
27. Latasa C, Solano C, Penadés JR, Lasa I. Biofilm-associated proteins. CR Biol. 2006; 329(11): 849-857.
28. Diani M, Esiyok OG, Ariafar MN, Yuksel FN, Altuntas EG, Akcelik N. The interactions between esp, fsr, gelE genes and biofilm formation and pfge analysis of clinical Enterococcus faecium strains. Afr J Microbiol Res. 2014; 8: 129-137.
29. Dworniczek E, Wojciech Ł, Sobieszczańska B, Seniuk A. Virulence of Enterococcus isolates collected in Lower Silesia (Poland). Scand J Infect Dis. 2005; 37: 630-636.
30. 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 E. faecium. Acta Biochim Pol. 2015; 62: 859-866.
31. Billström H, Sullivan A, Lund B. Cross-transmission of clinical Enterococcus faecium in relation to esp and antibiotic resistance. J Appl Microbiol. 2008; 105: 2115-2122.
32. Contreras GA, Diaz Granados CA, Cortes L, Reyes J, Vanegas S, et al. Nosocomial outbreak of Enterococcus gallinarum: untaming of rare species of enterococci. J Hosp Infect. 2008; 70: 346-352.
33. Franiczek R, Dolna I, Dworniczek E, Krzyżanowska B, Seniuk A, Piątkowska E. In vitro activity of tigecycline against clinical isolates of Gram-positive and Gram-negative bacteria displaying different resistance phenol-types. Adv Clin Exp Me. 2008; 17: 545-551.
34. Freitas AR, Coque TM, Novais C, Hammerum AM, Lester CH, Zervos MJ, et al. Human and swine hosts share vancomycin-resistant Enterococcus faecium CC17 and CC5 and Enterococcus faecalis CC2 clonal clusters harboring Tn1546 on indistinguishable plasmids. J Clin Microbiol. 2011; 49: 925-931.
35. Sieńko A, Wieczorek P, Wieczorek A, Sacha P, Majewski P, Ojdana D, et al. Occurrence of high-level aminoglycoside resistance (HLAR) among Enterococcus species strains. Prog Health Sci. 2014; 4: 179-187.
36. Werner G, Grforer S, Fleige C, Witte W, Klare I. Tigecycline-resistant Enterococcus faecalis strain isolated from a German Intensive Care Unit Patient. J Antimicrob Chemother. 2008; 61: 1182-1183.
37. 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: 100-103.
38. Dupre I, Zanetti S, Schito AM, Fadda G, Sechi LA. Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected in Sardinia (Italy). J Med Microbiol. 2003; 52: 491-498.
39. Tendolkar PM, Baghdayan AS, Gilmore MS, Shankar N. Enterococcal surface protein, Esp, enhances biofilm formation by Enterococcus faecalis. Infect Imm. 2004; 72: 6032-6039.
40. Hallgren A, Claesson C, Saeedi B, Monstein HJ, Hanberger H, Nilsson LE. Molecular detection of aggregation substance, enterococcal surface protein, and cytolysin genes and in vitro adhesion to urinary catheters of E. faecalis and E. faecium. Int J Med Microbiol. 2009; 299: 323-332.
41. Raad II, Hanna HA, Boktour M, Chaiban G, Hachem RY, Dvorak T, et al. Vancomycin-resistant Enterococcus faecium: catheter colonization, esp gene, and decreased susceptibility to antibiotics in biofilm. Antimicrob Agents Chemother. 2005; 49: 5046-5050.
42. 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: 4473-4479.
43. 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: 590-595.
44. Archimbaud C, Shankar N, Forestier C, Baghdayan A, Gilmore MS, Charbonne F, et al. In vitro adhesive properties and virulence factors of Enterococcus faecalis strains. Res Microbiol. 2002; 153: 75-80.
Published
2017-12-31
How to Cite
(1)
Sieńko, A.; Ojdana, D.; Majewski, P.; Sacha, P.; Wieczorek, P.; Tryniszewska, E. Comparison of Biofilm-Producing Enterococcus Faecalis, Enterococcus Faecium, and Unusual Enterococcus Strains. European Journal of Biological Research 2017, 7, 291-298.
Section
Research Articles
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