A commercially available microfluidics flow cell was utilized together with widefield fluorescence microscopy to evaluate the effects of disinfectants on bacterial strains. The flow cell’s inner surface supports the formation of biofilms of numerous bacterial species. The modular setup of the flow cell accessories allows connection to syringes, pumps and collection vials, facilitating aseptic experiments in a controlled fluidics environment which can be documented with precisely timed microscopy imaging. The flow cell is inoculated with a suspension of bacteria in a nutrient medium and incubated for several days allowing bacterial cells to form a biofilm. Shortly before performing an assay, the biofilm is labelled with a dual-fluorescent DNA probe which distinguishes unharmed and damaged bacteria. Then a disinfectant sample (or control) is gently injected and time-lapse imaging is used for quantifying the course of bacterial biomass response. We use a simplified widefield microscopy method that allows intensive recording and quantification of time series of two-dimensional frames for tracking the course of disinfectant action on a variety of microbial strains. This procedure has potential for the rapid evaluation of novel products.
Summary
A 71-year-old woman presented with acute non-cardiogenic pulmonary oedema. She proved to have a Pasteurella multocida pneumonia, with blood stream invasion by the organism, and required positive pressure ventilation for 53 days.
Penicillin G., the drug of choice for this infection, failed to reverse the steady decline in her arterial oxygen-tension, and it was only after treatment with chloramphenicol and prednisolone that she began to improve.
Serological tests strongly indicated the presence of a Staphylococcus aureus infection and the delay in giving antibiotics appropriate to this second pathogen may have been the reason for the patient’s initial downhill course.
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