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, Proteomic analysis of A. baumannii SDF in biofilms: highlighting atypical adhesion

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, Biofilm dispersion activity of UFAs was investigated only on 24 h-biofilms formed by ATCC 17978 strain in MHB. Twenty four-hour biofilms were incubated in fresh medium for an additional 24 h in the presence of fatty acids (at 0.01, 0.02, or 0.05 mg/mL) using DMSO or virstatin 100 µM as controls, case of strains forming pellicle in addition to biofilms on plate walls, the sub-phase was gently removed allowing the pellicle to stick onto the plate walls, then the overall remaining biomass (i.e., biofilm on solid surface and pellicle) was quantified, as described by, vol.38

, The effect of the addition of long-HSLs, i.e., N-(3-hydroxydodecanoyl)-DL-homoserine lactone (OH-C 12-HSL, Sigma Aldrich) and N-decanoyl-DL-homoserine lactone (N-C 10-HSL, Sigma Aldrich) solubilized in DMSO, on biofilms pretreated by virstatin and UFAs was investigated. Five hundred nM of each HSL were added concomitantly to 0.02 mg/mL of UFAs and 100 µM virstatin. DMSO enriched with HSL (500 nM) was used as control. Biofilm formation was quantified and the results significance was analyzed, as described in the previous section. The effect of virstatin and UFAs on abaR gene expression using q-RT-PCR was also investigated. A. baumannii was grown overnight in MH medium and diluted to OD600 of 0.01 and incubated for 24 h under agitation at 37 ? C with DMSO (negative control), virstatin (100 µM), MoA (0.02 mg/mL), or PoA (0.02 mg/mL). Total RNAs were extracted using RNeasy-Mini kit, Effect of Virstatin and UFAs on Quorum Sensing The N-acyl-homoserine lactone (AHL) production in A. baumannii ATCC 17978 or A. baumannii clinical isolates was determined, as described by [28] with P. aeruginosa PAO1 and E. coli ATCC 10536 as positive and negative controls, respectively. To investigate the potential quorum-quenching activity of UFAs and virstatin on short-HSLs, we used the screening platform described by, vol.29

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