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Background: Acinetobacter baumannii and Pseudomonas aeruginosa are of major concern for hospitalized patients., Methods: We evaluated antibiotic and antiseptic resistance of A. baumannii (n = 29) and P. aeruginosa (n = 37) isolates recovered from 66 intensive care unit (ICU) patients and determined the prevalence of qacE, qacEΔ1, and integrons in these clinical isolates. Antibiotic and antiseptic susceptibility testing was performed via Kirby Bauer disk diffusion and broth microdilution methods, respectively. The resistance genes and integrons were detected by PCR. A. baumannii and P. aeruginosa ICU isolates showed 100% and 70.3% antibiotic multiple drug resistance patterns, respectively., Results: The isolates also revealed high levels of resistance (MIC ≥ 16 µg/ml) against antiseptics commonly used in Egyptian hospitals (Benzalkonium, Benzethonium, and Chlorhexidine). The qacEΔ1 gene showed higher levels of prevalence in both A. baumannii and P. aeruginosa isolates (93.5% and 78%, respectively) as compared to that of qacE gene (52.0% and 33.0%, respectively). The intI1 was more prevalent among A. baumannii isolates (65.5%) compared to P. aeruginosa isolates (37.8%). P. aeruginosa resistance genotypes were significantly associated with antibiotic and antiseptic resistance patterns. A. baumannii resistance genotypes were associated with antiseptic-resistance patterns., Conclusion: The excessive usage of antiseptics may escalate bacterial resistance, especially with high prevalence of intI1 integron in these pathogens., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The increasing occurrence of resistance among Pseudomonas aeruginosa clinical isolates necessitates finding alternatives to antibiotics for controlling the infection of such pathogenic bacteria. In this study, lactonase gene ahl-1 from Bacillus weihenstephanensis isolate-P65 was successfully cloned and expressed in Escherichia coli BL21 (DE3) under the control of T7 promoter for utilizing its quorum quenching activity against three multidrug-resistant (MDR) P. aeruginosa clinical isolates. The biological activity of the overexpressed lactonase enzyme (Ahl-1), tested using a synthetic signal and Chromobacterium violaceum CV026 as a biosensor, displayed good catalytic activity using hexanoyl homoserine lactone (HHL) as a substrate and Chromobacterium violaceum (CV026) as a biosensor (77.2 and 133 nm min -1 for the crude and the purified Ahl-lactonase enzymes, respectively). Upon challenging its ability to inhibit the virulence of three MDR P. aeruginosa clinical isolates, recombinant Ahl-1 successfully prevented the accumulation of acylhomoserine lactone signals resulting in a significant reduction in the investigated virulence determinants; protease (from 40 up to 75.5%), pyocyanin (48-75.9%), and rhamnolipids (52.7-63.4%) (P value < 0.05). Ahl-1 also displayed significant inhibitory activities on the swarming motility and biofilm formation of the three tested MDR P. aeruginosa clinical isolates (P value < 0.05). Consequently, Ahl-1 lactonase enzyme in this study is considered a promising therapeutic agent to inhibit P. aeruginosa pathogenicity with no fear of emergence of resistance.

Pseudomonas aeruginosa colonizes the lungs in cystic fibrosis (CF) and mechanically ventilated patients by binding to the cellular receptors on the surface of the lung epithelium. Studies have shown that blocking this interaction could be achieved with sub-minimum inhibitory concentrations of antibiotics such as ciprofloxacin. The development of bacterial resistance is a probable drawback of such an intervention. The use of natural extracts to interfere with bacterial adhesion and invasion has recently gained substantial attention and is hypothesized to inhibit bacterial binding and consequently prevent or reduce pathogenicity. This study used an A549 lung epithelial cell infection model, and the results revealed that a combination of aqueous cranberry extract with ciprofloxacin could completely prevent the adhesion and invasion of P. aeruginosa PAO1 compared to the untreated control. All of the natural extracts (cranberry, dextran, and soybean extracts) and ciprofloxacin showed a significant reduction (P<0.0001) in P. aeruginosa PAO1 adhesion to and invasion of lung epithelial cells relative to the control. The cranberry, dextran, and soybean extracts could substantially increase the anti-adhesion and anti-invasion effects of ciprofloxacin to the averages of 100% (P<0.0001), 80% (P<0.0001), and 60% (P<0.0001), respectively. Those extracts might result in a lower rate of the development of bacterial resistance; they are relatively safe and inexpensive agents, and utilizing such extracts, alone or in combination with ciprofloxacin, as potential anti-adhesion and anti-invasion remedies, could be valuable in preventing or reducing P. aeruginosa lung infections., (Copyright © 2014 King Saud Bin Abdulaziz University for Health Sciences. Published by Elsevier Ltd. All rights reserved.)

Objectives: The conventional in vitro models simulate pharmacodynamics of antibiotics in the treatment of planktonic Pseudomonas aeruginosa. In this study, we propose a novel pharmacodynamic model of ofloxacin activity in the treatment of P. aeruginosa biofilm., Methods: P. aeruginosa biofilm carrying coupons were suspended in a continuous flow central compartment bioreactor (CCB). In the CCB, the pharmacokinetics of different ofloxacin dosing regimens were simulated. Samples from the coupons and the CCB were assessed for viability of the biofilm and the shedding planktonic cells, respectively, over 24h. In addition, ofloxacin concentrations were assessed in each sample withdrawn for the CCB using bioassay method., Results: The microbiological outcomes on P. aeruginosa biofilm and the shedding planktonic cells in response to different ofloxacin dosing regimens were not parallel and this may explain the non-coincidence of microbiological and clinical outcomes with biofilm associated infections., Conclusion: The current study has introduced unprecedented novel dynamic model for the assessment of the microbiological outcome on both biofilm and shedding planktonic cells of P. aeruginosa in response to different dosing regimens of ofloxacin which in turn can simulate the clinical outcomes in biofilm associated infections of P. aeruginosa, e.g. cystic fibrosis. Furthermore, different scenarios of antibiotic dosing regimens against biofilm related infections can be mimicked using such model.

The increase of multi-drug resistant Pseudomonas aeruginosa infections is a worldwide dilemma. At the heart of the problem is the inability to treat established P. aeruginosa biofilms with standard antibiotic therapy, including fluoroquinolones. We address a previously unstudied question as to the effect of a commonly prescribed calcium channel blocker (CCB) diltiazem on the biofilm growth. Real-time monitoring of the overall growth and killing of P. aeruginosa biofilm during fluoroquinolones therapy in the presence and absence of diltiazem was performed. In this study, we demonstrate that for P. aeruginosa biofilms, resistance to the first-line fluoroquinolones may be induced by the commonly prescribed calcium channel blocker diltiazem., (Published by Elsevier Ltd.)

A new method was developed to rapidly monitor the Pseudomonas aeruginosa viable counts using alamar blue (AB). The 96-well microtiter plates were used to perform the assay. This procedure is based on fluorogenic measurement as a result of reduction of nonfluorescent AB to red fluorescent form by the viable cells of P. aeruginosa. The correlation between conventional plate count and fluorogenic AB method was highly satisfactory for quantification of planktonic (R2 = 0.9487) and biofilm cells of P. aeruginosa (R2 = 0.9296). PRACTICAL APPLICATIONS The new fluorogenic method can rapidly monitor Pseudomonas aeruginosa counts in vitro with a high correlation with the conventional plating method. The results indicate that fluorogenic method requires much shorter time (2 h) than the conventional plate count (24 h), is a more cost-effective way, quite amenable to high throughput, and continuous monitoring of P. aeruginosa viability is achievable in the kinetic in vitro models without interference with the cell viability. [ABSTRACT FROM AUTHOR]