Abstract
BackgroundBiofilm formation is a major virulence factor contributing to the chronicity of infections. To date few studies have evaluated biofilm formation in infecting isolates of patients including both Gram-positive and Gram-negative multidrug-resistant (MDR) species in the context of numerous types of infectious syndromes. Herein, we investigated the biofilm forming capacity in a large collection of single patient infecting isolates and compared the relationship between biofilm formation to various strain characteristics.MethodsThe biofilm-forming capacity of 205 randomly sampled clinical isolates from patients, collected from various anatomical sites, admitted for treatment at Brooke Army Medical Center (BAMC) from 2004–2011, including methicillin-resistant/methicillin susceptible Staphylococcus aureus (MRSA/MSSA) (n=23), Acinetobacter baumannii (n=53), Pseudomonas aeruginosa (n=36), Klebsiella pneumoniae (n=54), and Escherichia coli (n=39), were evaluated for biofilm formation using the high-throughput microtiter plate assay and scanning electron microscopy (SEM). Relationships between biofilm formation to clonal type, site of isolate collection, and MDR phenotype were evaluated. Furthermore, in patients with relapsing infections, serial strains were assessed for their ability to form biofilms in vitro.ResultsOf the 205 clinical isolates tested, 126 strains (61.4%) were observed to form biofilms in vitro at levels greater than or equal to the Staphylococcus epidermidis, positive biofilm producing strain, with P. aeruginosa and S. aureus having the greatest number of biofilm producing strains. Biofilm formation was significantly associated with specific clonal types, the site of isolate collection, and strains positive for biofilm formation were more frequently observed to be MDR. In patients with relapsing infections, the majority of serial isolates recovered from these individuals were observed to be strong biofilm producers in vitro.ConclusionsThis study is the first to evaluate biofilm formation in a large collection of infecting clinical isolates representing diverse types of infections. Our results demonstrate: (1) biofilm formation is a heterogeneous property amongst clinical strains which is associated with certain clonal types, (2) biofilm forming strains are more frequently isolated from non-fluid tissues, in particular bone and soft tissues, (3) MDR pathogens are more often biofilm formers, and (4) strains from patients with persistent infections are positive for biofilm formation.
Highlights
Biofilm formation is a major virulence factor contributing to the chronicity of infections
Multidrug-resistant (MDR) organisms, including Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus (MRSA), and extended spectrum beta-lactamase (ESBL) producing Gram-negative bacteria, are frequently implicated as the causative agents of acute and chronic infections contributing significantly to patient morbidity and mortality, as well as increased health care costs associated with treatment [1,2]
With the exception of S. aureus, which was cultured in tryptic soy broth (TSB), all bacteria were grown in Luria-Bertani broth (LB) overnight at 37°C
Summary
Biofilm formation is a major virulence factor contributing to the chronicity of infections. To date few studies have evaluated biofilm formation in infecting isolates of patients including both Gram-positive and Gram-negative multidrug-resistant (MDR) species in the context of numerous types of infectious syndromes. Multidrug-resistant (MDR) organisms, including Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus (MRSA), and extended spectrum beta-lactamase (ESBL) producing Gram-negative bacteria, are frequently implicated as the causative agents of acute and chronic infections contributing significantly to patient morbidity and mortality, as well as increased health care costs associated with treatment [1,2]. Numerous studies to date indicate that human infections are, in large part, caused by the ability of bacteria to develop surface attached polymicrobial communities known as biofilms [3,4,5]. Biofilm formation is implicated as a significant factor involved in a number of chronic human infections [4,16,17]
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