1887

Abstract

Biofilms plays an important role in medical-device-related infections. This study aimed to determine the factors that influence adherence and biofilm production, as well as the relationship between strong biofilm production and genetic determinants in clinical isolates of meticillin-resistant (MRSA). Fifteen strains carrying different chromosomal cassettes recovered from hospitalized patients were selected; five SCCII, five SCCIII and five SCCIV. The SCC type, group and the presence of the virulence genes ( and IS) were assessed by PCR. PFGE and multilocus sequence typing (MLST) techniques were also performed. The initial adhesion and biofilm formation were examined by quantitative assays. The surface tension and hydrophobicity of the strains were measured by the contact angle technique to evaluate the association between these parameters and adhesion ability. SCCIII and IV strains were less hydrophilic, with a high value for the electron acceptor parameter and higher adhesion in comparison with SCCII strains. Only SCCIII strains could be characterized as strong biofilm producers. The PFGE showed five major pulsotypes (A–E); however, biofilm production was related to the dissemination of one specific PFGE clone (C) belonging to MLST ST239 (Brazilian epidemic clonal complex). The genes I, and IS in SCCIII strains were considered as genetic determinants associated with strong biofilm-formation by an -independent biofilm pathway. This study contributes to the understanding of biofilm production as an aggravating factor potentially involved in the persistence and severity of infections caused by multidrug-resistant MRSA belonging to this genotype.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000228
2016-04-01
2019-10-21
Loading full text...

Full text loading...

/deliver/fulltext/jmm/65/4/286.html?itemId=/content/journal/jmm/10.1099/jmm.0.000228&mimeType=html&fmt=ahah

References

  1. Amaral M. M. , Coelho L. R. , Flores R. P. , Souza R. R. , Silva-Carvalho M. C. , Teixeira L. A. , Ferreira-Carvalho B. T. , Figueiredo A. M. . ( 2005;). The predominant variant of the Brazilian epidemic clonal complex of methicillin-resistant Staphylococcus aureus has an enhanced ability to produce biofilm and to adhere to and invade airway epithelial cells. J Infect Dis 192: 801–810 [CrossRef] [PubMed].
    [Google Scholar]
  2. Arciola C. R. , Baldassarri L. , Montanaro L. . ( 2001;). Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections. J Clin Microbiol 39: 2151–2156 [CrossRef] [PubMed].
    [Google Scholar]
  3. Azeredo J. , Oliveira R. . ( 2000;). The role of exopolymers in the attachment of Sphingomonas paucimobilis . Biofouling 16: 59–67.[CrossRef]
    [Google Scholar]
  4. Cafiso V. , Bertuccio T. , Santagati M. , Demelio V. , Spina D. , Nicoletti G. , Stefani S. . ( 2007;). agr-Genotyping and transcriptional analysis of biofilm-producing Staphylococcus aureus . FEMS Immunol Med Microbiol 51: 220–227.[CrossRef]
    [Google Scholar]
  5. Carvalho K. S. , Mamizuka E. M. , Gontijo Filho P. P. . ( 2010;). Methicillin/oxacillin-resistant Staphylococcus aureus as a hospital and public health threat in Brazil. Braz J Infect Dis 14: 71–76 [CrossRef] [PubMed].
    [Google Scholar]
  6. Cassat J. E. , Smeltzer M. S. , Lee C. Y. . ( 2014;). Investigation of biofilm formation in clinical isolates of Staphylococcus aureus . Methods Mol Biol 1085: 195–211 [CrossRef] [PubMed].
    [Google Scholar]
  7. Cerca N. , Pier G. B. , Vilanova M. , Oliveira R. , Azeredo J. . ( 2005;). Quantitative analysis of adhesion and biofilm formation on hydrophilic and hydrophobic surfaces of clinical isolates of Staphylococcus epidermidis . Res Microbiol 156: 506–514 [CrossRef] [PubMed].
    [Google Scholar]
  8. Cha J. O. , Yoo J. I. , Yoo J. S. , Chung H. S. , Park S. H. , Kim H. S. , Lee Y. S. , Chung G. T. . ( 2013;). Investigation of biofilm formation and its association with the molecular and clinical characteristics of methicillin-resistant Staphylococcus aureus . Osong Public Health Res Perspect 4: 225–232 [CrossRef] [PubMed].
    [Google Scholar]
  9. Chaignon P. , Sadovskaya I. , Ragunah Ch. , Ramasubbu N. , Kaplan J. B. , Jabbouri S. . ( 2007;). Susceptibility of staphylococcal biofilms to enzymatic treatments depends on their chemical composition. Appl Microbiol Biotechnol 75: 125–132 [CrossRef] [PubMed].
    [Google Scholar]
  10. Chambers H. F. , Deleo F. R. . ( 2009;). Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol 7: 629–641 [CrossRef] [PubMed].
    [Google Scholar]
  11. CLSI ( 2013;). Performance Standards for Antimicrobial Susceptibility Testing; 23rd Informational Supplement M100-S23. Wayne, PA: Clinical and Laboratory Standards Institute;.
  12. Costa M. O. , Beltrame C. O. , Ferreira F. A. , Botelho A. M. , Lima N. C. , Souza R. C. , de Almeida L. G. , Vasconcelos A. T. , Nicolás M. F. , Figueiredo A. M. . ( 2013;). Complete genome sequence of a variant of the methicillin-resistant Staphylococcus aureus ST239 lineage, strain BMB9393, displaying superior ability to accumulate ica-independent biofilm. Genome Announc 1: e00576–13 [CrossRef] [PubMed].
    [Google Scholar]
  13. Cucarella C. , Solano C. , Valle J. , Amorena B. , Lasa I. , Penadés J. R. . ( 2001;). Bap, a Staphylococcus aureus surface protein involved in biofilm formation. J Bacteriol 183: 2888–2896 [CrossRef] [PubMed].
    [Google Scholar]
  14. Cucarella C. , Tormo M. A. , Ubeda C. , Trotonda M. P. , Monzón M. , Peris C. , Amorena B. , Lasa I. , Penadés J. R. . ( 2004;). Role of biofilm-associated protein bap in the pathogenesis of bovine Staphylococcus aureus . Infect Immun 72: 2177–2185.[CrossRef]
    [Google Scholar]
  15. Doyle R. J. . ( 2000;). Contribution of the hydrophobic effect to microbial infection. Microbes Infect 2: 391–400 [CrossRef] [PubMed].
    [Google Scholar]
  16. Enright M. C. , Day N. P. , Davies C. E. , Peacock S. J. , Spratt B. G. . ( 2000;). Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus . J Clin Microbiol 38: 1008–1015 [PubMed].
    [Google Scholar]
  17. Ferreira F. A. , Souza R. R. , de Sousa Moraes B. , de Amorim Ferreira A. M. , Américo M. A. , Fracalanzza S. E. , Dos Santos Silva Couceiro J. N. , Sá Figueiredo A. M. . ( 2013;). Impact of agr dysfunction on virulence profiles and infections associated with a novel methicillin-resistant Staphylococcus aureus (MRSA) variant of the lineage ST1-SCCmecIV. BMC Microbiol 13: 93 [CrossRef] [PubMed].
    [Google Scholar]
  18. Foster T. J. , Geoghegan J. A. , Ganesh V. K. , Höök M. . ( 2014;). Adhesion, invasion and evasion: the many functions of the surface proteins of Staphylococcus aureus . Nat Rev Microbiol 12: 49–62 [CrossRef] [PubMed].
    [Google Scholar]
  19. Gales A. C. , Sader H. S. , Ribeiro J. , Zoccoli C. , Barth A. , Pignatari A. C. . ( 2009;). Antimicrobial susceptibility of gram-positive bacteria isolated in Brazilian hospitals participating in the SENTRY Program (2005-2008). Braz J Infect Dis 13: 90–98.
    [Google Scholar]
  20. Geoghegan J. A. , Corrigan R. M. , Gruszka D. T. , Speziale P. , O'Gara J. P. , Potts J. R. , Foster T. J. . ( 2010;). Role of surface protein SasG in biofilm formation by Staphylococcus aureus . J Bacteriol 192: 5663–5673 [CrossRef] [PubMed].
    [Google Scholar]
  21. George E. A. , Muir T. W. . ( 2007;). Molecular mechanisms of agr quorum sensing in virulent staphylococci. ChemBioChem 8: 847–855 [CrossRef] [PubMed].
    [Google Scholar]
  22. Goering R. V. . ( 2010;). Pulsed field gel electrophoresis: a review of application and interpretation in the molecular epidemiology of infectious disease. Infect Genet Evol 10: 866–875.[CrossRef]
    [Google Scholar]
  23. Gordon R. J. , Lowy F. D. . ( 2008;). Pathogenesis of methicillin-resistant Staphylococcus aureus infection. Clin Infect Dis 46: (Suppl. 5), S350–S359 [CrossRef] [PubMed].
    [Google Scholar]
  24. Gu J. , Li H. , Li M. , Vuong C. , Otto M. , Wen Y. , Gao Q. . ( 2005;). Bacterial insertion sequence IS256 as a potential molecular marker to discriminate invasive strains from commensal strains of Staphylococcus epidermidis . J Hosp Infect 61: 342–348 [CrossRef] [PubMed].
    [Google Scholar]
  25. Hiramatsu K. , Ito T. , Tsubakishita S. , Sasaki T. , Takeuchi F. , Morimoto Y. , Katayama Y. , Matsuo M. , Kuwahara-Arai K. , other authors . ( 2013;). Genomic basis for methicillin resistance in Staphylococcus aureus . Infect Chemother 45: 117–136 [CrossRef] [PubMed].
    [Google Scholar]
  26. Katsikogianni M. , Missirlis Y. F. . ( 2004;). Concise review of mechanisms of bacterial adhesion to biomaterials and of techniques used in estimating bacteria-material interactions. Eur Cell Mater 8: 37–57 [PubMed].
    [Google Scholar]
  27. Kondo Y. , Ito T. , Ma X. X. , Watanabe S. , Kreiswirth B. N. , Etienne J. , Hiramatsu K. . ( 2007;). Combination of multiplex PCRs for staphylococcal cassette chromosome mec type assignment: rapid identification system for mec, ccr, and major differences in junkyard regions. Antimicrob Agents Chemother 51: 264–274 [CrossRef] [PubMed].
    [Google Scholar]
  28. Kwon A. S. , Park G. C. , Ryu S. Y. , Lim D. H. , Lim D. Y. , Choi C. H. , Park Y. , Lim Y. . ( 2008;). Higher biofilm formation in multidrug-resistant clinical isolates of Staphylococcus aureus . Int J Antimicrob Agents 32: 68–72 [CrossRef] [PubMed].
    [Google Scholar]
  29. Laverty G. , Gorman S. P. , Gilmore B. F. . ( 2013;). Biomolecular mechanisms of staphylococcal biofilm formation. Future Microbiol 8: 509–524 [CrossRef] [PubMed].
    [Google Scholar]
  30. Lim Y. , Shin H. J. , Kwon A. S. , Reu J. H. , Park G. , Kim J. . ( 2013;). Predictive genetic risk markers for strong biofilm-forming Staphylococcus aureus: fnbB gene and SCCmec type III. Diagn Microbiol Infect Dis 76: 539–541 [CrossRef] [PubMed].
    [Google Scholar]
  31. Lyon B. R. , Gillespie M. T. , Skurray R. A. . ( 1987;). Detection and characterization of IS256, an insertion sequence in Staphylococcus aureus . J Gen Microbiol 133: 3031–3038 [PubMed].
    [Google Scholar]
  32. Magiorakos A. P. , Srinivasan A. , Carey R. B. , Carmeli Y. , Falagas M. E. , Giske C. G. , Harbarth S. , Hindler J. F. , Kahlmeter G. , other authors . ( 2012;). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 18: 268–281 [CrossRef] [PubMed].
    [Google Scholar]
  33. Martín-López J. V. , Díez-Gil O. , Morales M. , Batista N. , Villar J. , Claverie-Martín F. , Méndez-Alvarez S. . ( 2004;). Simultaneous PCR detection of ica cluster and methicillin and mupirocin resistance genes in catheter-isolated Staphylococcus . Int Microbiol 7: 63–66.
    [Google Scholar]
  34. McCourt J. , O'Halloran D. P. , McCarthy H. , O'Gara J. P. , Geoghegan J. A. . ( 2014;). Fibronectin-binding proteins are required for biofilm formation by community-associated methicillin-resistant Staphylococcus aureus strain LAC. FEMS Microbiol Lett 353: 157–164 [CrossRef] [PubMed].
    [Google Scholar]
  35. McDougal L. K. , Steward C. D. , Killgore G. E. , Chaitram J. M. , McAllister S. K. , Tenover F. C. . ( 2003;). Pulsed-field gel electrophoresis typing of oxacillin-resistant Staphylococcus aureus isolates from the United States: establishing a national database. J Clin Microbiol 41: 5113–5120.[CrossRef]
    [Google Scholar]
  36. McEvoy C. R. , Tsuji B. , Gao W. , Seemann T. , Porter J. L. , Doig K. , Ngo D. , Howden B. P. , Stinear T. P. . ( 2013;). Decreased vancomycin susceptibility in Staphylococcus aureus caused by IS256 tempering of WalKR expression. Antimicrob Agents Chemother 57: 3240–3249 [CrossRef] [PubMed].
    [Google Scholar]
  37. Merino N. , Toledo-Arana A. , Vergara-Irigaray M. , Valle J. , Solano C. , Calvo E. , Lopez J. A. , Foster T. J. , Penadés J. R. , Lasa I. . ( 2009;). Protein A-mediated multicellular behavior in Staphylococcus aureus . J Bacteriol 191: 832–843 [CrossRef] [PubMed].
    [Google Scholar]
  38. Montanaro L. , Speziale P. , Campoccia D. , Ravaioli S. , Cangini I. , Pietrocola G. , Giannini S. , Arciola C. R. . ( 2011;). Scenery of Staphylococcus implant infections in orthopedics. Future Microbiol 6: 1329–1349 [CrossRef] [PubMed].
    [Google Scholar]
  39. O'Gara J. P. . ( 2007;). ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus . FEMS Microbiol Lett 270: 179–188 [CrossRef] [PubMed].
    [Google Scholar]
  40. O'Neill E. , Pozzi C. , Houston P. , Humphreys H. , Robinson D. A. , Loughman A. , Foster T. J. , O'Gara J. P. . ( 2008;). A novel Staphylococcus aureus biofilm phenotype mediated by the fibronectin-binding proteins, FnBPA and FnBPB. J Bacteriol 190: 3835–3850 [CrossRef] [PubMed].
    [Google Scholar]
  41. O'Toole G. A. . ( 2011;). Microtiter dish biofilm formation assay. J Vis Exp 47: 2437 [PubMed].
    [Google Scholar]
  42. Otto M. . ( 2013;). Staphylococcal infections: mechanisms of biofilm maturation and detachment as critical determinants of pathogenicity. Annu Rev Med 64: 175–188 [CrossRef] [PubMed].
    [Google Scholar]
  43. Rodríguez-Noriega E. , Seas C. . ( 2010;). The changing pattern of methicillin-resistant Staphylococcus aureus clones in Latin America: implications for clinical practice in the region. Braz J Infect Dis 14: (Suppl. 2), S87–S96 [CrossRef] [PubMed].
    [Google Scholar]
  44. Rodríguez-Noriega E. , Seas C. , Guzmán-Blanco M. , Mejía C. , Alvarez C. , Bavestrello L. , Zurita J. , Labarca J. , Luna C. M. , other authors . ( 2010;). Evolution of methicillin-resistant Staphylococcus aureus clones in Latin America. Int J Infect Dis 14: e560–e566 [CrossRef] [PubMed].
    [Google Scholar]
  45. Sader H. S. , Jones R. N. . ( 2009;). Antimicrobial susceptibility of Gram-positive bacteria isolated from US medical centers: results of the Daptomycin Surveillance Program (2007–2008). Diagn Microbiol Infect Dis 65: 158–162 [CrossRef] [PubMed].
    [Google Scholar]
  46. Sader H. S. , Moet G. J. , Jones R. N. . ( 2009;). Antimicrobial resistance among Gram-positive bacteria isolated in Latin American hospitals. J Chemother 21: 611–620 [CrossRef] [PubMed].
    [Google Scholar]
  47. Schlievert P. M. , Strandberg K. L. , Lin Y. C. , Peterson M. L. , Leung D. Y. . ( 2010;). Secreted virulence factor comparison between methicillin-resistant and methicillin-sensitive Staphylococcus aureus, and its relevance to atopic dermatitis. J Allergy Clin Immunol 125: 39–49 [CrossRef] [PubMed].
    [Google Scholar]
  48. Schroeder K. , Jularic M. , Horsburgh S. M. , Hirschhausen N. , Neumann C. , Bertling A. , Schulte A. , Foster S. , Kehrel B. E. , other authors . ( 2009;). Molecular characterization of a novel Staphylococcus aureus surface protein (SasC) involved in cell aggregation and biofilm accumulation. PLoS One 4: e7567 [CrossRef] [PubMed].
    [Google Scholar]
  49. Shopsin B. , Mathema B. , Alcabes P. , Said-Salim B. , Lina G. , Matsuka A. , Martinez J. , Kreiswirth B. N. . ( 2003;). Prevalence of agr specificity groups among Staphylococcus aureus strains colonizing children and their guardians. J Clin Microbiol 41: 456–459 [CrossRef] [PubMed].
    [Google Scholar]
  50. Sousa C. , Teixeira P. , Oliveira R. . ( 2009;). Influence of surface properties on the adhesion of Staphylococcus epidermidis to acrylic and silicone. Int J Biomater 2009: 718017 [PubMed].[CrossRef]
    [Google Scholar]
  51. Speziale P. , Pietrocola G. , Foster T. J. , Geoghegan J. A. . ( 2014;). Protein-based biofilm matrices in staphylococci. Front Cell Infect Microbiol 4: 171 [PubMed].[CrossRef]
    [Google Scholar]
  52. Stepanović S. , Vuković D. , Hola V. , Di Bonaventura G. , Djukić S. , Cirković I. , Ruzicka F. . ( 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 [CrossRef] [PubMed].
    [Google Scholar]
  53. Strandberg K. L. , Rotschafer J. H. , Vetter S. M. , Buonpane R. A. , Kranz D. M. , Schlievert P. M. . ( 2010;). Staphylococcal superantigens cause lethal pulmonary disease in rabbits. J Infect Dis 202: 1690–1697 [CrossRef] [PubMed].
    [Google Scholar]
  54. Toledo-Arana A. , Merino N. , Vergara-Irigaray M. , Débarbouillé M. , Penadés J. R. , Lasa I. . ( 2005;). Staphylococcus aureus develops an alternative, ica-independent biofilm in the absence of the arlRS two-component system. J Bacteriol 187: 5318–5329 [CrossRef] [PubMed].
    [Google Scholar]
  55. Tristan A. , Ying L. , Bes M. , Etienne J. , Vandenesch F. , Lina G. . ( 2003;). Use of multiplex PCR to identify Staphylococcus aureus adhesins involved in human hematogenous infections. J Clin Microbiol 41: 4465–4467 [CrossRef] [PubMed].
    [Google Scholar]
  56. van der Mei H. S. , Bos R. , Busscher H. J. . ( 1998;). A reference guide to microbial surface hydrophobicity based on contact angles. Colloids Surf B Biointerfaces 11: 213–221 [CrossRef].
    [Google Scholar]
  57. van Oss C. J. . ( 1997;). Hydrophobicity and hydrophilicity of biosurfaces. Curr Opin Colloid Interface Sci 2: 503–512 [CrossRef].
    [Google Scholar]
  58. van Oss C. J. , Giese R. F. . ( 1995;). The hydrophilicity and hydrophobicity of clay minerals. Clays Clay Miner 43: 474–477 [CrossRef].
    [Google Scholar]
  59. Vautor E. , Abadie G. , Pont A. , Thiery R. . ( 2008;). Evaluation of the presence of the bap gene in Staphylococcus aureus isolates recovered from human and animals species. Vet Microbiol 127: 407–411 [CrossRef] [PubMed].
    [Google Scholar]
  60. Vergara-Irigaray M. , Valle J. , Merino N. , Latasa C. , García B. , Ruiz de Los Mozos I. , Solano C. , Toledo-Arana A. , Penadés J. R. , Lasa I. . ( 2009;). Relevant role of fibronectin-binding proteins in Staphylococcus aureus biofilm-associated foreign-body infections. Infect Immun 77: 3978–3991 [CrossRef] [PubMed].
    [Google Scholar]
  61. Vuong C. , Saenz H. L. , Götz F. , Otto M. . ( 2000;). Impact of the agr quorum-sensing system on adherence to polystyrene in Staphylococcus aureus . J Infect Dis 182: 1688–1693 [CrossRef] [PubMed].
    [Google Scholar]
  62. Wu Z. , Li F. , Liu D. , Xue H. , Zhao X. . ( 2015;). Novel type XII staphylococcal cassette chromosome mec harboring a new cassette chromosome recombinase. CcrC2. Antimicrob Agents Chemother 59: 7597–7601 [CrossRef] [PubMed].
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000228
Loading
/content/journal/jmm/10.1099/jmm.0.000228
Loading

Data & Media loading...

Supplements

Supplementary Data



PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error