Comparative In Vitro Study of Biofilm Formation and Antimicrobial Susceptibility in Gram-Negative Bacilli Isolated from Prosthetic Joint Infections

Prosthetic joint infections (PJIs) are typically caused by microorganisms that grow in biofilms. Traditional antimicrobial susceptibility tests are based on the study of planktonic bacteria that might lead to missing the biofilm behavior and to a treatment failure. This study was designed to analyze the antimicrobial susceptibility of clinical Gram-negative bacilli (GNB) isolates from PJIs in planktonic and sessile states and the possible relationship between antimicrobial resistance and biofilm formation. A total of 46 clinical isolates from patients with PJIs (mainly hip and knee prostheses) plus three GNB ATCC isolates were studied. The Minimal Inhibitory Concentration (MIC), minimal bactericidal concentration (MBC), minimal biofilm inhibitory concentration (MBIC), and minimal biofilm eradication concentration (MBEC) were assessed using a previously published methodology. Almost all of the GNB clinical isolates tested were biofilm forming. Pseudomonas aeruginosa was the largest biofilm-forming species. A comparison of MBIC(90) versus MIC(90) shows an increase higher than 1- to -2-fold dilutions in most antimicrobials studied, and MBEC90 was significantly higher than MIC90, becoming resistant to all the antimicrobial drugs tested. Higher biofilm production values were obtained in antibiotic-susceptible Escherichia coli in comparison to their resistant counterparts. However, regarding the relationships between antimicrobial resistance and biofilm formation, our analysis showed that each strain differed. A high antimicrobial resistance rate was found among the GNB studied. Moreover, almost all bacterial isolates were in vitro biofilm formers. Although there was no significant association between biofilm and antibiotic resistance, multidrug-resistant isolates were found to be greater biofilm formers than non-multidrug-resistant isolates. IMPORTANCE This study is the first one to analyze a high number of isolates of Gram-negative bacilli that are the cause of prosthetic joint infection. The analysis includes biofilm development and antimicrobial susceptibility testing of both planktonic and sessile bacteria. The obtained results support the clinical knowledge about the treatment of these bacteria when biofilms are involved.