Living cells of Staphylococcus aureus immobilized onto the capillary surface in electrochromatography: a tool for screening of biofilms.
Microorganisms attach to nonliving surfaces in many natural, industrial, and medical environments, enveloped within extracellular polymeric substances. The result is a biofilm. Biofilms are reported to exist in 65-80% of bacterial infections refractory to host defenses and antibiotics therapy and are regarded as a central problem in present-day medical microbiology. Understanding of the parameters governing the interaction of antimicrobials with biofilms is thus of great interest in any attempt to increase biocide efficacy. In this work, study was made of the feasibility of using open tubular capillary electrochromatography (CEC) in bacterial biofilm studies with living cells. Staphylococcus aureus was selected as model bacterium. First, S. aureus was shown, under various conditions, to form a biofilm on the inner wall of a fused-silica capillary coated with poly(L-lysine). Optimal conditions for biofilm formation, such as bacterial concentration, growing time, and the stability of the ensemble, were preliminarily defined with conventional 96-microtiter well plates. Continuous flushing of the capillary with fresh cells meant that no growth medium was needed. The presence of biofilm in the capillary was confirmed by atomic force microscopy. Interactions between S. aureus biofilms and different antibiomicrobial agents were studied by capillary electrochromatography. The effect of five antibiotics (penicillin G, oxacillin, fusidic acid, rifampicin, vancomycin) on biofilms was examined in terms of retention factors and reduced mobilities of the antibiotics. The antibiotic susceptibility profile for S. aureus is similar as the result of minimal inhibitory concentrations registered on the 96-microtiter well plates for both planktonic and biofilm cells. The results show, for the first time, that bacterial biofilms can be studied by CEC. The technique allows highly efficient and easy characterization of interactions between S. aureus biofilms and potentially active antimicrobial compounds under different conditions. Reagent and cell consumption are minimal.
Chen
Jie
J
Laboratory of Analytical Chemistry, Department of Chemistry, P.O. Box 55, University of Helsinki, FIN-00014 Helsinki, Finland.
Fallarero
Adyary
A
Määttänen
Anni
A
Sandberg
Malena
M
Peltonen
Jouko
J
Vuorela
Pia M
PM
Riekkola
Marja-Liisa
ML
D018829:Adhesins, Bacterial; D000890:Anti-Infective Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects; D053801:Capillary Electrochromatography / Q000379:methods*; D005240:Feasibility Studies; D008826:Microbial Sensitivity Tests / Q000379:methods; D018625:Microscopy, Atomic Force; D011107:Polylysine / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D013211:Staphylococcus aureus / Q000187:drug effects / Q000502:physiology*
D018829:Adhesins, Bacterial; D000890:Anti-Infective Agents; D011107:Polylysine; D012822:Silicon Dioxide