Stimulation and inhibition of bacterial growth by caffeine dependent on chloramphenicol and a phenolic uncoupler--a ternary toxicity study using microfluid segment technique.
A droplet-based microfluidic technique for the fast generation of three dimensional concentration spaces within nanoliter segments was introduced. The technique was applied for the evaluation of the effect of two selected antibiotic substances on the toxicity and activation of bacterial growth by caffeine. Therefore a three-dimensional concentration space was completely addressed by generating large sequences with about 1150 well separated microdroplets containing 216 different combinations of concentrations. To evaluate the toxicity of the ternary mixtures a time-resolved miniaturized optical double endpoint detection unit using a microflow-through fluorimeter and a two channel microflow-through photometer was used for the simultaneous analysis of changes on the endogenous cellular fluorescence signal and on the cell density of E. coli cultivated inside 500 nL microfluid segments. Both endpoints supplied similar results for the dose related cellular response. Strong non-linear combination effects, concentration dependent stimulation and the formation of activity summits on bolographic maps were determined. The results reflect a complex response of growing bacterial cultures in dependence on the combined effectors. A strong caffeine induced enhancement of bacterial growth was found at sublethal chloramphenicol and sublethal 2,4-dinitrophenol concentrations. The reliability of the method was proved by a high redundancy of fluidic experiments. The results indicate the importance of multi-parameter investigations for toxicological studies and prove the potential of the microsegmented flow technique for such requirements.
Ilmenau University of Technology, Institute of Micro- and Nano Technologies/Institute of Chemistry and Biotechnology, Department of Physical Chemistry and Micro Reaction Technology, Ilmenau, 98693, Germany.
Jialan
J
Cao
Dana
D
Kürsten
Steffen
S
Schneider
J Michael
JM
Köhler
D019297:2,4-Dinitrophenol / Q000494:pharmacology*; D002110:Caffeine / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D002701:Chloramphenicol / Q000494:pharmacology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000187:drug effects*; D017022:Flow Injection Analysis / Q000295:instrumentation; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D019800:Phenol / Q000037:antagonists & inhibitors*; D018675:Toxicity Tests / Q000295:instrumentation*
D019800:Phenol; D002110:Caffeine; D002701:Chloramphenicol; D019297:2,4-Dinitrophenol