Evaluation of membrane fluidity of multidrug-resistant isolates of Escherichia coli and Staphylococcus aureus in presence and absence of antibiotics.

In the face of the serious problem of antimicrobial resistance and the global dissemination of multidrug-resistant (MDR) bacteria, it is relevant to deeply study such bacteria both genetically and phenotypically. It is well known that bacteria have the ability to modify the biophysical properties of their cytoplasmic membranes, namely fluidity, in order to survive and thrive in hostile environments. The aim of this study was to assess and compare the membrane fluidity among multidrug-resistant (MDR) isolates of Escherichia coli and Staphylococcus aureus in absence and in presence of antibiotics (ceftazidime or ciprofloxacin). The membrane fluidity was monitored at 24-h intervals up to three days and at the sixth day, by measuring the anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH) and the generalized polarization (GP) of Laurdan (6-dodecanoyl-2-dimethylaminonaphthalene). The anisotropy values as well as the Laurdan excitation GP (GPexc) values obtained from all three E. coli strains (two MDR isolates and one susceptible reference strain) were quite similar and indicative of a very alike membrane phospholipid composition, regardless harboring or not resistance to multiple antimicrobials. Nonetheless, in the case of S. aureus, the anisotropy values were more increased in methicillin-resistant S. aureus (MRSA) isolates in comparison to the reference strain, meaning they have a less fluid membrane. Equally, GPexc values were statistically different among the three S. aureus strains and showed that the two MRSA isolates had more rigid membranes than the susceptible strain. The exposition of MDR isolates of S. aureus to subinhibitory concentrations of ciprofloxacin did not affect neither the anisotropy values nor the GPexc values, therefore, not altering membrane fluidity. Membrane integrity, assessed by the Live/Dead staining, of all strains and conditions studied was maintained over the six days. Thus, these preliminary studies on membrane properties of MDR isolates demonstrate that i) MRSA seem to have a more rigid membrane that susceptible S. aureus and ii) the presence of subinhibitory concentrations of antibiotics does not significantly alter the membrane fluidity of S. aureus, regardless being MDR or susceptible, but slightly affect the membrane fluidity of E. coli.