Comparison of antibiotic resistance and virulence between biofilm-producing and non-producing clinical isolates of Enterococcus faecium

• Authors: Anna Sieńko , Piotr Wieczorek , Piotr Majewski , Dominika Ojdana , Anna Wieczorek , Dorota Olszańska , Elżbieta Tryniszewska
• Languages of publication: EN

Abstracts

EN

An increase in the antibiotic resistance among Enterococcus faecium strains has been observed worldwide. Moreover, this bacteria has the ability to produce several virulence factors and to form biofilm that plays an important role in human infections. This study was designed to compare the antibiotic resistance and the prevalence of genes encoding surface protein (esp), aggregation substance (as), surface adhesin (efaA), collagen adhesin (ace), gelatinase (gelE), and hialuronidase (hyl) between biofilm-producing and non-producing E. faecium strains. Therefore, ninety E. faecium clinical isolates were tested for biofilm-forming ability, and then were assigned to two groups: biofilm-positive (BIO+, n =70) and biofilm-negative (BIO-, n = 20). Comparison of these groups showed that BIO+ isolates were resistant to β-lactams, whereas 10% of BIO- strains were susceptible to ampicillin (statistically significant difference, p = 0.007) and 5% to imipenem. Linezolid and tigecycline were the only antibiotics active against all tested isolates. Analysis of the virulence factors revealed that ace, efaA, and gelE genes occurred more frequently in BIO- strains (ace in 50% BIO+ vs. 75% BIO-; efaA 44.3% vs. 85%; gelE 2.9% vs. 15%, respectively), while hyl gene appeared more frequently in BIO+ isolates (87.1% BIO+ vs. 65% BIO-). These differences were significant (p < 0.05). We concluded that BIO+ strains were more resistant to antibiotics than BIO- strains, but interestingly, BIO- isolates were characterized by possession of higher virulence capabilities.

Keywords

EN

Enterococcus; biofilm; virulence; resistance

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2015
• Volume: 62
• Issue: 4
• Pages: 859-866

Dates

published

2015

received

2015-08-01

revised

2015-09-18

accepted

2015-10-05

(unknown)

2015-12-04

Contributors

author

Anna Sieńko

• Department of Microbiological Diagnostics and Infectious Immunology, Medical University of Bialystok, Bialystok, Poland

author

Piotr Wieczorek

• Department of Microbiological Diagnostics and Infectious Immunology, Medical University of Bialystok, Bialystok, Poland

author

Piotr Majewski

• Department of Microbiological Diagnostics and Infectious Immunology, Medical University of Bialystok, Bialystok, Poland

author

Dominika Ojdana

• Department of Microbiological Diagnostics and Infectious Immunology, Medical University of Bialystok, Bialystok, Poland

author

Anna Wieczorek

• Department of Microbiological Diagnostics and Infectious Immunology, Medical University of Bialystok, Bialystok, Poland

author

Dorota Olszańska

• Department of Microbiological Diagnostics and Infectious Immunology, University Hospital of Bialystok, Bialystok, Poland

author

Elżbieta Tryniszewska

• Department of Microbiological Diagnostics and Infectious Immunology, Medical University of Bialystok, Bialystok, Poland
• Department of Microbiological Diagnostics and Infectious Immunology, University Hospital of Bialystok, Bialystok, Poland

References

• Akhter J, Ahmed S, Saleh AA, Anwar S (2014) Antimicrobial resistance and in vitro biofilm-forming ability of Enterococci spp. isolated from urinary tract infection in a tertiary care hospital in Dhaka. Bangladesh Med Res Counc Bull 40: 6-9.
• Almohamad S, Somarajan SR, Singh KV, Nallapareddy SR, Murray BE (2014) Influence of isolate origin and presence of various genes on biofilm formation by Enterococcus faecium. FEMS Microbiol Lett 353: 151-156.
• Amyes SG (2007) Enterococci and streptococci. Int J Antimicrob Agents 29 (Suppl 3): S43-S52.
• Baldir G, Engin DO, Kucukercan M, Inan A, Akcay S, Ozyuker S, Asaray S (2013) High-level resistance to aminoglycoside, vancomycin and linezolid in enterococci strains. J Microbiol Infect Dis 3: 100-103.
• Billström H, Sullivan A, Lund B (2008) Cross-transmission of clinical Enterococcus faecium in relation to esp and antibiotic resistance. J Appl Microbiol 105: 2115-2122.
• Bronk M, Samet A (2008) Szpitalne bakteriemie enterokokowe. Post Mikrob 47: 339-344.
• Cabrera-Contreras R, Morelos-Ramirez R, Galicia-Camacho AN, Melendez-Herrada E (2013) Antibiotic resistance and biofilm production in Staphylococcus epidermidis strains, isolated from a Tertiary Care Hospital in Mexico City. ISRN Microbiol 2013: 918921.
• Camargo IL, Gilmore MS, Darini AL (2006) Multilocus sequence typing and analysis of putative virulence factors in vancomycin-resistant and vancomycin-sensitive Enterococcus faecium isolates from Brazil. Clin Microbiol Infect 12: 1123-1130.
• Chai WL, Hamimah H, Cheng SC, Sallam AA, Abdullah M (2007) Susceptibility of Enterococcus faecalis biofilm to antibiotics and calcium hydroxide. J Oral Sci 49: 161-166.
• Chen AY, Zerros JM (2009) Enterococcus: Anti-microbial Resistance in Enterococci. Epidemiology, Treatment and Control. Antimicrobial Drug Resistance and Infectious Disease 49: 715-733. Humana Press.
• Cheng V, Chen J, Tai J, Wong S, Poon R, Hung I, To K, Chan J, Ho PL, Lo CM, Yuen KY (2014) Decolonization of gastrointestinal carriage of vancomycin-resistant Enterococcus faecium: case series and review of literature. BMC Infect Dis 14: 514.
• Christensen GD, Simpson WA, Bisno AL, Beachey EH (1982) Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun 37: 318-326.
• Comerlato CB, Resende MC, Caierão J, d'Azevedo PA (2013) Presence of virulence factors in Enterococcus faecalis and Enterococcus faecium susceptible and resistant to vancomycin. Mem Inst Oswaldo Cruz 108: 590-595.
• Diani M, Esiyok OG, Ariafar MN, Yuksel FN, Altuntas EG, Akcelik N (2014) The interactions between esp, fsr, gelE genes and biofilm formation and pfge analysis of clinical Enterococcus faecium strains. Afr J Microbiol Res 8: 129-137.
• Di Rosa R, Creti R, Venditti M, D'Amelio R, Arciola CR, Montanaro L, Baldassarri L (2006) Relationship between biofilm formation, the enterococcal surface protein (Esp) and gelatinase in clinical isolates of Enterococcus faecalis and Enterococcus faecium. FEMS Microbiol Lett 256: 145-150.
• Dupre I, Zanetti S, Schito AM, Fadda G, Sechi LA (2003) Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected in Sardinia (Italy). J Med Microbiol 52: 491-498.
• Dutka-Malen S, Evers S, Courvalin P (1995) Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR. J Clin Microbiol 33: 24-27.
• Dworniczek E, Wojciech Ł, Sobieszczańska B, Seniuk A (2005) Virulence of Enterococcus isolates collected in Lower Silesia (Poland). Scand J Infect Dis 37: 630-636.
• European Centre for Disease Prevention and Control, Antimicrobial resistance surveillance in Europe 2013. ECDC, Stockholm, Sweden, 2014.
• Fisher K, Phillips C (2009) The ecology, epidemiology and virulence of Enterococcus. Microbiology 155: 1749-1757.
• Franiczek R, Dolna I, Dworniczek E, Krzyżanowska B, Seniuk A, Piątkowska E (2008) In vitro activity of Tigecycline against clinical isolates of Gram-positive and Gram-negative bacteria displaying different resistance phenol-types. Adv Clin Exp Me 17: 545-551.
• Frank KL, Vergidis P, Brinkman CL, Greenwood Quaintance KE, Barnes AM, Mandrekar JN, Schlievert PM, Dunny GM, Patel R (2015) Evaluation of the Enterococcus faecalis biofilm-associated virulence factors AhrC and Eep in rat foreign body osteomyelitis and in vitro biofilm-associated antimicrobial resistance. PLoS One 10: e0130187.
• Freeman DJ, Falkiner FR, Keane CT (1989) New method for detecting slime production by coagulase negative staphylococci. J Clin Pathol 42: 872-874.
• Freitas AR, Coque TM, Novais C, Hammerum AM, Lester CH, Zervos MJ, Donabedian S, Jensen LB, Francia MV, Baquero F, Peixe L (2011) 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 49: 925-931.
• Hancock LE, Perego M (2004) The Enterococcus faecalis fsr two-component system controls biofilm development through production of gelatinase. J Bacteriol 186: 5629-5639.
• Hasani A, Sharifi Y, Ghotaslou R, Naghili B, Hasani A, Aghazadeh M, Milani M, Bazmani A (2012) 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 30: 175-181.
• Heikens E, Bonten MJ, Willems RJ (2007) Enterococcal surface protein Esp is important for biofilm formation of Enterococcus faecium E1162. J Bacteriol 189: 8233-8240.
• Heikens E, Singh KV, Jacques-Palaz KD, van Luit-Asbroek M, Oostdijk EA, Bonten MJ, Murray BE, Willems RJ (2011) Contribution of the enterococcal surface protein Esp to pathogenesis of Enterococcus faecium endocarditis. Microbes Infect 13: 1185-1190.
• Hryniewicz W, Wardal E (2009) Enterokoki oporne na wankomycynę - groźny patogen zakażeń szpitalnych. Nowa Klin 7: 711-716 (in Polish).
• Iris N, Sayıner HS, Yıldırmak T, Şimşek, F, Arat ME (2014) Distribution of vancomycin resistant enterococci and their resistance patterns determined by surveillance. Afr J Microbiol Res 8: 680-684.
• Kafil HS, Mobarez AM (2015) Assessment of biofilm formation by enterococci isolates from urinary tract infections with different virulence profiles. J King Saud Univ Sci
• Kowalska-Krochmal B, Dworniczek E, Dolna I, Bania J, Wałecka E, Seniuk A, Gościniak G (2011) Resistance patterns and occurrence of virulence determinants among GRE strains in southwestern Poland. Adv Med Sci 56: 304-310.
• Latasa C, Solano C, Penadés JR, Lasa I (2006) Biofilm-associated proteins. C R Biol 329: 849-857.
• Lewis K (2001) Riddle of Biofilm Resistance. Antimicrob Agents Chemother 45: 999-1007.
• Lins RX, de Oliveira Andrade A, Hirata Junior R, Wilson MJ, Lewis MA, Williams DW, Fidel RA (2013) Antimicrobial resistance and virulence traits of Enterococcus faecalis from primary endodontic infections. J Dent 41: 779-786.
• Mohamed JA, Huang DB (2007) Biofilm formation by enterococci. J Med Microbiol 56: 1581-1588.
• Oliveira A, Cunha MD (2010) Comparison of methods for the detection of biofilm production in coagulase-negative staphylococci. BMC Res Notes 3: 260.
• Özden Tuncer B, Ay Z, Tuncer Y (2013) Occurrence of enterocin genes, virulence factors, and antibiotic resistance in 3 bacteriocin-producer Enterococcus faecium strains isolated from Turkish tulum cheese. Turk J Biol 37: 443-449.
• Paganelli FL, Willems RJ, Jansen P, Hendrickx A, Zhang X, Bonten MJ, Leavis HL (2013) Enterococcus faecium biofilm formation: identification of major autolysin AtlAEfm, associated Acm surface localization, and AtlAEfm-independent extracellular DNA Release. MBio 4: e00154.
• Pitkälä A, Salmikivi L, Bredbacka P, Myllyniemi AL, Koskinen MT (2007) Comparison of tests for detection of beta-lactamase-producing staphylococci. J Clin Microbiol 45: 2031-2033.
• Praharaj I, Sujatha S, Parija SC (2013) Phenotypic & genotypic characterization of vancomycin resistant Enterococcus isolates from clinical specimens. Indian J Med Res 138: 549-556.
• Prakash VP, Rao SR, Parija SC (2005) Emergence of unusual species of enterococci causing infections, South India. BMC Infect Dis 5: 14.
• Raad II, Hanna HA, Boktour M, Chaiban G, Hachem RY, Dvorak T, Lewis R, Murray BE (2005) Vancomycin-resistant Enterococcus faecium: Catheter colonization, esp gene, and decreased susceptibility to antibiotics in biofilm. Antimicrob Agents Chemother 49: 5046-5050.
• Rodriguez-Bano J, Ramirez E, Muniain MA, Santos J, Joyanes P, Gonzalez F, Garcia-Sanchez M, Martinez-Martinez L (2005) Colonization by high-level aminoglycoside-resistant enterococci in intensive care unit patients: epidemiology and clinical relevance. J Hosp Infect 60: 353-359.
• Sacha P, Wieczorek P, Jakoniuk P (2008) Oporność na wankomycynę wśród szczepów Enterococcus faecium. Przegl lek 65: 229-232 (in Polish).
• Sava IG, Heikens E, Huebner J (2010) Pathogenesis and immunity in enterococcal infections. Clin Microbiol Infect 16: 533-540.
• Schouten MA, Voss A, Hoogkamp-Korstanje JA (1999) Antimicrobial susceptibility patterns of Enterococci causing infections in Europe. The European VRE Study Group. Antimicrob Agents Chemother 43: 2542-2546.
• Sieńko A, Wieczorek P, Wieczorek A, Sacha P, Majewski P, Ojdana D, Michalska A, Tryniszewska E (2014) Occurrence of high-level aminoglycoside resistance (HLAR) among Enterococcus species strains. Prog Health Sci 4: 179-187.
• Simonsen GS, Småbrekke L, Monnet DL, Sørensen TL, Møller JK, Kristinsson KG, Lagerqvist-Widh A, Torell E, Digranes A, Harthug S, Sundsfjord A (2003) 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 51: 323-331.
• Top J, Willems R, Blok H, de Regt M, Jalink K, Troelstra A, Goorhuis B, Bonten M (2007) Ecological replacement of Enterococcus faecalis by multiresistant clonal complex 17 Enterococcus faecium. Clin Microbiol Infect 13: 316-319.
• Vakulenko SB, Donabedian SM, Voskresenskiy AM, Zervos MJ, Lerner SA, Chow JW (2003) Multiplex PCR for detection of aminoglycoside resistance genes in Enterococci. Antimicrob Agents Chemother 47: 1423-1426.
• Vankerckhoven V, Van Autgaerden T, Vael C, Lammens C, Chapelle S, Rossi R, Jabes D, Goossens H (2004) 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 42: 4473-4479.
• Werner G, Gfrorer S, Fleige C, Witte W, Klare I (2008) Tigecycline-resistant Enterococcus faecalis strain isolated from a German Intensive Care Unit Patient. J Antimicrob Chemother 61: 1182-1183.
• Wu D, Fan W, Kishen A, Gutmann JL, Fan B (2014) Evaluation of the antibacterial efficacy of silver nanoparticles against Enterococcus faecalis biofilm. J Endod 40: 285-290.
• Yayanthi S, Ananthasubramanian M, Appalaraju B (2008) Assessment of pheromone response in biofilm forming clinical isolates of high level gentamicin resistant Enterococcus faecalis. Indian J Med Microbiol 26: 248-251.
• Zou LK, Wang HN, Zeng B, Li JN, Li XT, Zhang AY, Zhou YS, Yang X, Xu CW, Xia QQ (2011) Erythromycin resistance and virulence genes in Enterococcus faecalis from swine in China. New Microbiol 34: 73-80.

Identifiers

DOI

10.18388/abp.2015_1147