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J Med Microbiol 57 (2008), 1018-1023; DOI: 10.1099/jmm.0.2008/000968-0
© 2008 Society for General Microbiology
ISSN 1473-5644

Biofilm formation by Scottish clinical isolates of Staphylococcus aureus

Karen Smith1, Ana Perez1, Gordon Ramage2,{dagger}, David Lappin2, Curtis G. Gemmell3 and Sue Lang1,{dagger}

1 Department of Biological and Biomedical Sciences, Glasgow Caledonian University, Glasgow, UK

2 Section of Infection and Immunity, Glasgow Dental School, University of Glasgow, Glasgow, UK

3 Honorary Professor of Microbiology, Universities of Glasgow and St Andrews, UK

Correspondence
Sue Lang
sue.lang{at}gcal.ac.uk

Received 4 February 2008
Accepted 22 April 2008

The biofilm-forming capacity of 972 clinical isolates of Staphylococcus aureus was tested using a high-throughput polystyrene 96-peg plate format. Isolates of S. aureus were collected from patients in hospitals throughout Scotland from 2004 to 2006; 763 of these were meticillin-resistant S. aureus (MRSA) and 209 were meticillin-sensitive S. aureus (MSSA). The biomass of each biofilm was quantified using a crystal violet staining technique. Isolates were divided into those that formed fully established biofilms, moderately attached biofilms and weakly adherent biofilms by comparison with a known biofilm-forming strain. The majority of MRSA (53.8 %) and MSSA (43.5 %) isolates formed moderately attached biofilms. Fully established biofilms were formed by 20.5 % of MRSA isolates and 28.0 % of MSSA isolates, whilst 25.7 % of MRSA isolates and 28.5 % of MSSA isolates formed negligible biofilms. <-- INSERT SHAPE --><-- INSERT SHAPE -->There was no significant correlation between susceptibility to meticillin and biofilm formation (P=0.77). MRSA isolates were divided into clonal types (EMRSA-15, EMRSA-16 and sporadic isolates) based on PFGE genotyping results. EMRSA-15 isolates formed significantly more moderately and fully established biofilms than EMRSA-16 isolates (P<0.001). S. aureus strains isolated from the skin of patients had a significantly greater capacity to form biofilms than isolates from other body sites, including the blood. Microscopic examination of biofilms by scanning electron microscopy (SEM) revealed that poorly adherent biofilm formers failed to colonize the entire surface of the peg, whilst moderately adherent biofilm formers grew in uniform monolayers but failed to develop a mature three-dimensional structure. SEM analysis of an isolate representative of the group that formed fully established biofilms confirmed that this isolate developed a dense biofilm with a textured, multi-layered, three-dimensional structure.

Abbreviations: EMRSA, epidemic MRSA; MRSA, meticillin-resistant Staphylococcus aureus; MSSA, meticillin-sensitive S. aureus; SEM, scanning electron microscopy; SMRSARL, Scottish MRSA Reference Laboratory.

{dagger}These authors contributed equally to this work.






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