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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.

In this investigation, a GT1-7 cell-based cytotoxicity screening assay in 96-well microplates was set up. The assay, using propidium iodide fluorescence, was proven to be reliable, with good quality (Z' = 0.51) and low plate-to-plate and day-to-day variations. Further on, a library containing extracts from 227 genetic modification (GM) Gerbera hybrida and 42 Gerbera varieties was screened; however, no differences between them were found. Based on these findings, we propose the use of the current assay within the first-tier screening studies of large collections. Also, these results provide valuable information for GM Gerbera risk-assessment purposes and offer a model for the toxicity cell-based screening of GM crops.