Research article

Biofilm synthesis and its relationship with genetic characteristics in clinical methicillin-resistant staphylococci

  • Received: 29 June 2015 Accepted: 25 September 2015 Published: 29 September 2015
  • Staphylococcus aureus can cause a broad range of infections, including skin infections, pneumonia and bacteraemia. Coagulase-negative staphylococci (CNS), mainly S. epidermidis, have also emerged as important pathogens, especially in immunocompromised patients or those with prosthetic devices, such as intravascular catheters or biomaterials. Of great importance in the initiation of these infections is the ability of staphylococci to adhere to various surfaces, such as host tissues and prosthetic devices and to form biofilm. The staphylococcal adhesins are encoded by a number of genes such as fnbA (S. aureus fibronectin binding protein A), sasG (S. aureus surface protein G), aap (S. epidermidis accumulation associated protein), bhp (Bap homologue protein) and fbe (fibrinogen binding protein epidermidis). In this study, 106 methicillin-resistant S. aureus (MRSA), 145 methicillin-resistant S. epidermidis (MRSE) and 70 non-epidermidis methicillin-resistant CNS (MR-CNS; 58 S. haemolyticus, 10 S. hominis and two S. lugdunensis) were compared in terms of biofilm formation, antimicrobial resistance, clonal distribution and adhesin genes carriage. Isolates were classified into pulsotypes by PFGE and assigned to sequence types by MLST. In total, 121/321 isolates (37.7%) produced biofilm and 219 (68.2%) carried ica operon. The majority was multidrug resistant (94.7%) and carried one or more adhesin genes. MRSE and all other MR-CNS prevailed in biofilm formation (P < 0.001) and antimicrobial resistance (P < 0.05) as compared to MRSA. MRSE also prevailed in ica carriage compared to the other methicillin-resistant staphylococci (P ≤ 0.007) Among MRSE, isolates from bacteraemias prevailed in biofilm formation (P = 0.031), whereas, strains from prosthetic device-associated infections carried more frequently aap (P = 0.003). Even though PFGE showed genetic diversity among MRSE, MLST revealed three major clones (ST2, ST5, ST16). MRSA isolates were less diverse, with five PFGE types and, among them, one major PFGE type (C) consisting of 77/106 strains (72.6%). MLST identified five sequence types: ST5, ST30, ST80, ST225 and ST239. One major PFGE type (h) was identified in S. haemolyticus. A clonal relationship was found concerning fnbA carriage in MRSA, ica carriage in MRSE, and antimicrobial susceptibility in both groups reinforcing the aspect of clonal expansion in hospital settings.

    Citation: Nikolaos Giormezis, Konstantinos Papakonstantinou, Fevronia Kolonitsiou, Eleanna Drougka, Antigoni Foka, Styliani Sarrou, Evangelos D. Anastassiou, Efthimia Petinaki, Iris Spiliopoulou. Biofilm synthesis and its relationship with genetic characteristics in clinical methicillin-resistant staphylococci[J]. AIMS Bioengineering, 2015, 2(4): 375-386. doi: 10.3934/bioeng.2015.4.375

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  • Staphylococcus aureus can cause a broad range of infections, including skin infections, pneumonia and bacteraemia. Coagulase-negative staphylococci (CNS), mainly S. epidermidis, have also emerged as important pathogens, especially in immunocompromised patients or those with prosthetic devices, such as intravascular catheters or biomaterials. Of great importance in the initiation of these infections is the ability of staphylococci to adhere to various surfaces, such as host tissues and prosthetic devices and to form biofilm. The staphylococcal adhesins are encoded by a number of genes such as fnbA (S. aureus fibronectin binding protein A), sasG (S. aureus surface protein G), aap (S. epidermidis accumulation associated protein), bhp (Bap homologue protein) and fbe (fibrinogen binding protein epidermidis). In this study, 106 methicillin-resistant S. aureus (MRSA), 145 methicillin-resistant S. epidermidis (MRSE) and 70 non-epidermidis methicillin-resistant CNS (MR-CNS; 58 S. haemolyticus, 10 S. hominis and two S. lugdunensis) were compared in terms of biofilm formation, antimicrobial resistance, clonal distribution and adhesin genes carriage. Isolates were classified into pulsotypes by PFGE and assigned to sequence types by MLST. In total, 121/321 isolates (37.7%) produced biofilm and 219 (68.2%) carried ica operon. The majority was multidrug resistant (94.7%) and carried one or more adhesin genes. MRSE and all other MR-CNS prevailed in biofilm formation (P < 0.001) and antimicrobial resistance (P < 0.05) as compared to MRSA. MRSE also prevailed in ica carriage compared to the other methicillin-resistant staphylococci (P ≤ 0.007) Among MRSE, isolates from bacteraemias prevailed in biofilm formation (P = 0.031), whereas, strains from prosthetic device-associated infections carried more frequently aap (P = 0.003). Even though PFGE showed genetic diversity among MRSE, MLST revealed three major clones (ST2, ST5, ST16). MRSA isolates were less diverse, with five PFGE types and, among them, one major PFGE type (C) consisting of 77/106 strains (72.6%). MLST identified five sequence types: ST5, ST30, ST80, ST225 and ST239. One major PFGE type (h) was identified in S. haemolyticus. A clonal relationship was found concerning fnbA carriage in MRSA, ica carriage in MRSE, and antimicrobial susceptibility in both groups reinforcing the aspect of clonal expansion in hospital settings.


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    [1] Drougka E, Foka A, Liakopoulos A, et al. (2014) A 12-year survey of methicillin-resistant Staphylococcus aureus infections in Greece: ST80-IV epidemic? Clin Microbiol Infect 20: O796-803. doi: 10.1111/1469-0691.12624
    [2] McCann MT, Gilmore BF, Gorman SP (2008) Staphylococcus epidermidis device-related infections: pathogenesis and clinical management. J Pharm Pharmacol 60: 1551-1571. doi: 10.1211/jpp.60.12.0001
    [3] Santos Sanches I, Mato R, de Lencastre H, et al. (2000) Patterns of multidrug resistance among methicillin-resistant hospital isolates of coagulase-positive and coagulase-negative staphylococci collected in the international multicenter study RESIST in 1997 and 1998. Microb Drug Resist 6: 199-211. doi: 10.1089/mdr.2000.6.199
    [4] Hiramatsu K, Cui L, Kuroda M, et al. (2001) The emergence and evolution of methicillin-resistant Staphylococcus aureus. Trends Microbiol 9: 486-493. doi: 10.1016/S0966-842X(01)02175-8
    [5] Donlan RM, Costerton JW (2002) Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15: 167-193. doi: 10.1128/CMR.15.2.167-193.2002
    [6] Mack D, Fischer W, Krokotsch A, et al. (1996) The intercellular adhesin involved in biofilm accumulation of Staphylococcus epidermidis is a linear beta-1,6-linked glucosaminoglycan: purification and structural analysis. J Bacteriol 178: 175-183.
    [7] Tristan A, Ying L, Bes M, et al. (2003) Use of multiplex PCR to identify Staphylococcus aureus adhesins involved in human hematogenous infections. J Clin Microbiol 41: 4465-4467. doi: 10.1128/JCM.41.9.4465-4467.2003
    [8] Edwards AM, Potts JR, Josefsson E, et al. (2010) Staphylococcus aureus host cell invasion and virulence in sepsis is facilitated by the multiple repeats within FnBPA. PLoS Pathog 6: e1000964. doi: 10.1371/journal.ppat.1000964
    [9] Kuroda M, Ito R, Tanaka Y, et al. (2008) Staphylococcus aureus surface protein SasG contributes to intercellular autoaggregation of Staphylococcus aureus. Biochem Biophys Res Commun 377: 1102-1106. doi: 10.1016/j.bbrc.2008.10.134
    [10] Rohde H, Burdelski C, Bartscht K, et al. (2005) Induction of Staphylococcus epidermidis biofilm formation via proteolytic processing of the accumulation-associated protein by staphylococcal and host proteases. Mol Microbiol 55: 1883-1895. doi: 10.1111/j.1365-2958.2005.04515.x
    [11] Arciola CR, Campoccia D, Gamberini S, et al. (2004) Presence of fibrinogen-binding adhesin gene in Staphylococcus epidermidis isolates from central venous catheters-associated and orthopaedic implant-associated infections. Biomaterials 25: 4825-4829. doi: 10.1016/j.biomaterials.2003.11.056
    [12] Bowden MG, Chen W, Singvall J, et al. (2005) Identification and preliminary characterization of cell-wall-anchored proteins of Staphylococcus epidermidis. Microbiology 151: 1453-1464. doi: 10.1099/mic.0.27534-0
    [13] Roche FM, Meehan M, Foster TJ (2003) The Staphylococcus aureus surface protein SasG and its homologues promote bacterial adherence to human desquamated nasal epithelial cells. Microbiology 149: 2759-2767. doi: 10.1099/mic.0.26412-0
    [14] Hartford O, O'Brien L, Schofield K, et al. (2001) The Fbe (SdrG) protein of Staphylococcus epidermidis HB promotes bacterial adherence to fibrinogen. Microbiology 147: 2545-2552. doi: 10.1099/00221287-147-9-2545
    [15] Tormo MA, Knecht E, Gotz F, et al. (2005) Bap-dependent biofilm formation by pathogenic species of Staphylococcus: evidence of horizontal gene transfer? Microbiology 151: 2465-2475. doi: 10.1099/mic.0.27865-0
    [16] Horan TC, Andrus M, Dudeck MA (2008) CDC/NHSN surveillance definition of health care-associated infection and criteria for specific types of infections in the acute care setting. Am J Infect Control 36: 309-332. doi: 10.1016/j.ajic.2008.03.002
    [17] Kontos F, Petinaki E, Spiliopoulou I, et al. (2003) Evaluation of a novel method based on PCR Restriction Fragment Length Polymorphism Analysis of the tuf gene for the identification of Staphylococcus species. J Microbiol Methods 55: 465-469. doi: 10.1016/S0167-7012(03)00173-8
    [18] The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 4.0, 2014. http://www.eucast.org.
    [19] Stepanovic S, Vukovic D, Hola V, et al. (2007) Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 115: 891-899. doi: 10.1111/j.1600-0463.2007.apm_630.x
    [20] Murakami K, Minamide W, Wada K, et al. (1991) Identification of methicillin-resistant strains of staphylococci by polymerase chain reaction. J Clin Microbiol 29: 2240-2244.
    [21] Cafiso V, Bertuccio T, Santagati M, et al. (2004) Presence of the ica operon in clinical isolates of Staphylococcus epidermidis and its role in biofilm production. Clin Microbiol Infect 10: 1081-1088. doi: 10.1111/j.1469-0691.2004.01024.x
    [22] Gomes AR, Vinga S, Zavolan M, et al. (2005) Analysis of the genetic variability of virulence-related loci in epidemic clones of methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 49: 366-379. doi: 10.1128/AAC.49.1.366-379.2005
    [23] Vandecasteele SJ, Peetermans WE, R RM, et al. (2003) Reliability of the ica, aap and atlE genes in the discrimination between invasive, colonizing and contaminant Staphylococcus epidermidis isolates in the diagnosis of catheter-related infections. Clin Microbiol Infect 9: 114-119. doi: 10.1046/j.1469-0691.2003.00544.x
    [24] Giormezis N, Kolonitsiou F, Foka A, et al. (2014) Coagulase-negative staphylococcal bloodstream and prosthetic-device-associated infections: the role of biofilm formation and distribution of adhesin and toxin genes. J Med Microbiol 63: 1500-1508. doi: 10.1099/jmm.0.075259-0
    [25] Potter A, Ceotto H, Giambiagi-Demarval M, et al. (2009) The gene bap, involved in biofilm production, is present in Staphylococcus spp. strains from nosocomial infections. J Microbiol 47: 319-326.
    [26] Sandoe JA, Longshaw CM (2001) Ventriculoperitoneal shunt infection caused by Staphylococcus lugdunensis. Clin Microbiol Infect 7: 385-387. doi: 10.1046/j.1198-743x.2001.00268.x
    [27] Tenover FC, Arbeit RD, Goering RV, et al. (1995) Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 33: 2233-2239.
    [28] Miragaia M, Carrico JA, Thomas JC, et al. (2008) Comparison of molecular typing methods for characterization of Staphylococcus epidermidis: proposal for clone definition. J Clin Microbiol 46: 118-129. doi: 10.1128/JCM.01685-07
    [29] Wertheim HF, Melles DC, Vos MC, et al. (2005) The role of nasal carriage in Staphylococcus aureus infections. Lancet Infect Dis 5: 751-762. doi: 10.1016/S1473-3099(05)70295-4
    [30] Liakopoulos A, Spiliopoulou I, Damani A, et al. (2010) Dissemination of two international linezolid-resistant Staphylococcus epidermidis clones in Greek hospitals. J Antimicrob Chemother 65: 1070-1071. doi: 10.1093/jac/dkq065
    [31] Mertens A, Ghebremedhin B (2013) Genetic determinants and biofilm formation of clinical Staphylococcus epidermidis isolates from blood cultures and indwelling devises. Eur J Microbiol Immunol (Bp) 3: 111-119. doi: 10.1556/EuJMI.3.2013.2.4
    [32] Speziale P, Pietrocola G, Foster TJ, et al. (2014) Protein-based biofilm matrices in Staphylococci. Front Cell Infect Microbiol 4: 171.
    [33] Linke DGA (2011) Bacterial Adhesion. Chemistry, Biology and Physics: Springer Netherlands. 374 p.
    [34] Cucarella C, Solano C, Valle J, et al. (2001) Bap, a Staphylococcus aureus surface protein involved in biofilm formation. J Bacteriol 183: 2888-2896. doi: 10.1128/JB.183.9.2888-2896.2001
    [35] Corrigan RM, Rigby D, Handley P, et al. (2007) The role of Staphylococcus aureus surface protein SasG in adherence and biofilm formation. Microbiology 153: 2435-2446. doi: 10.1099/mic.0.2007/006676-0
    [36] Patel JD, Colton E, Ebert M, et al. (2012) Gene expression during S. epidermidis biofilm formation on biomaterials. J Biomed Mater Res A 100: 2863-2869.
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