Background and Aim: The emergence of colistin-resistant strains is considered a great threat for patients with severe infections. Here, we investigate the prevalence and some possible mechanisms of colistin resistance among multidrug-resistant (MDR) and extensively drug-resistant (XDR) Pseudomonas aeruginosa (P. aeruginosa). Methods: Antimicrobial susceptibility was performed using disc diffusion methods while colistin resistance was detected by agar dilution method. Possible mechanisms for colistin resistance were studied by detection of mcr-1 and mcr-2 genes by conventional PCR, detection of efflux mechanisms using Carbonyl Cyanide 3-Chlorophenylhydrazone (CCCP), studying outer membrane protein profile and Lipopolysaccharide (LPS) profile of resistant isolates. Results: It was found that MDR and XDR represented 96% and 87% of the isolated P. aeruginosa, respectively, and colistin resistance represented 21.3%. No isolates were positive for mcr-2 gene while 50% of colistin-resistant isolates were positive for mcr-1. Efflux mechanisms were detected in 3 isolates. Protein profile showed the presence of a band of 21.4 KDa in the resistant strains which may represent OprH while LPS profile showed differences among colistin-resistant mcr-1 negative strains, colistin-resistant mcr-1 positive strains and susceptible strains. Conclusion: The current study reports a high prevalence of colistin resistance and mcr-1 gene in P. aeruginosa strains isolated from Egypt that may result in untreatable infections. Our finding makes it urgent to avoid unnecessary clinical use of colistin.

Silver is a potent antimicrobial agent against a variety of microorganisms and once the element has entered the bacterial cell, it accumulates as silver nanoparticles with large surface area causing cell death. At the same time, the bacterial cell becomes a reservoir for silver. This study aims to test the microcidal effect of silver-killed E. coli O104: H4 and its supernatant against fresh viable cells of the same bacterium and some other species, including E. coli O157: H7, Multidrug Resistant (MDR) Pseudomonas aeruginosa and Methicillin Resistant Staphylococcus aureus (MRSA). Silver-killed bacteria were examined by Transmission Electron Microscopy (TEM). Agar well diffusion assay was used to test the antimicrobial efficacy and durability of both pellet suspension and supernatant of silver-killed E. coli O104:H4 against other bacteria. Both silver-killed bacteria and supernatant showed prolonged antimicrobial activity against the tested strains that extended to 40 days. The presence of adsorbed silver nanoparticles on the bacterial cell and inside the cells was verified by TEM. Silver-killed bacteria serve as an efficient sustained release reservoir for exporting the lethal silver cations. This promotes its use as a powerful disinfectant for polluted water and as an effective antibacterial which can be included in wound and burn dressings to overcome the problem of wound contamination.

Background: Pseudomonas aeruginosa (P. aeruginosa) represents a great threat to public health worldwide, due to its high ability to acquire resistance to different antibiotic classes. Carbapenems are effective against multidrug resistant (MDR) P. aeruginosa, but their widespread use has resulted in the emergence of carbapenem-resistant strains, which is considered a major global concern. This study aimed to determine the prevalence of carbapenem resistance among P. aeruginosa strains isolated from different sites of infection. Methods: Between October 2016 and February 2018, a total of 530 clinical specimens were collected from patients suffering from different infections, then processed and cultured. Isolates were tested for extended spectrum β-lactamase (ESBL) and metallo-β-lactamase (MBL) production using double-disk synergy test, modified Hodge tests, and disc potentiation test. PCR was used for the detection of selected OXA carbapenemases encoding genes. Results: Of 530 samples, 150 (28.3%) P. aeruginosa isolates were obtained. MDR strains were found in 66.6% (100 of 150) of isolates. Of 100 MDR P. aeruginosa isolates, 54 (54%) were ESBL producers and 21 (21%) carbapenem resistant P. aeruginosa. MBL production was found in 52.3% (eleven) carbapenem-resistant isolates. CTX-M15 was found among 55.5% of ESBL- producing P. aeruginosa. Carbapenemase genes detected were bla_IMP (42.8%, nine of 21), bla_VIM (52.3%, eleven of 21), bla_GIM (52.3%, eleven of 21), bla_SPM (38%, 8/21). In addition, isolates that were positive for the tested genes showed high resistance to other antimicrobials, such as colistin sulfate and tigecycline. Conclusion: Our study indicates that P. aeruginosa harboring ESBL and MBL with limited sensitivity to antibiotics are common among the isolated strains, which indicates the great problem facing the treatment of serious infectious diseases. As such, there is a need to study the resistance patterns of isolates and carry out screening for the presence of ESBL and MBL enzymes, in order to choose the proper antibiotic.

Background: Staphylococci are a common cause of catheter-associated urinary tract infections. The present study evaluated biofilm forming capacity and the presence of both icaA and icaD genes among staphylococci strains isolated from patients undergoing ureteral catheterization. Methodology: Different bacterial strains were isolated from urine and stents segments collected from 100 patients. Strains were identified by traditional microbiological methods. Stents were examined for biofilm using a scanning electron microscope (SEM). Staphylococcal isolates were tested for their ability to produce biofilm using the tissue culture plate assay method (TCP). The presence of icaA and icaD genes was determined by PCR technique. Results: Fifty-three staphylococcal strains were isolated and identified from 284 samples (18.7%). Forty-six staphylococcal strains were isolated from stent segment cultures while only seven strains were isolated from urine samples at the day of stent removal. S. aureus represented 6.3%, and S. epidermidis represented 12.3%. Out of the 18 S. aureus strains, 15 (83.3%) were biofilm producers and out of 35 S. epidermidis strains, 31 (88.6%) were biofilm producers. Staphylococcal strains were further classified as high (56.6%), moderate (30.2%) and non biofilm producers (13.2%). All biofilm producing strains were positive for icaA and icaD genes, and all biofilm negative strains were negative for both genes. Conclusion: Staphylococci isolated from catheter segments showed a higher extent of biofilm production than that isolated from urine samples. All biofilm producing staphylococci were positive for icaA and icaD genes, which indicates the important role of ica genes as virulence markers in staphylococcal infections associated with urinary catheterization.

Background: The rapid emergence of antimicrobial resistance among Gram-positive organisms, especially staphylococci, has become a serious clinical challenge. Efflux machinery and biofilm formation are considered two of the main causes of antimicrobial resistance and therapy failure. Aim: Our study aims to evaluate the antibiofilm and efflux pump inhibitory activity of the antifungal ketoconazole against multidrug-resistant (MDR) Staphylococcus aureus. Methods: Ketoconazole was tested for its effect on the following: minimum inhibitory concentrations (MICs) of ciprofloxacin, norfloxacin, levofloxacin, and ethidium bromide (EtBr) by the broth microdilution method, the efflux of EtBr by NorA-positive MDR S. aureus, and the relative expression of NorA, NorB, and NorC efflux pump genes. Docking studies of ketoconazole were performed using 1PW4 (glycerol-3-phosphate transporter from Escherichia coli which was the representative structure from the major facilitator superfamily). Results: Ketoconazole significantly decreased the MICs of levofloxacin, ciprofloxacin, norfloxacin, and EtBr (a substrate for efflux pump) by 8 to 1024-fold (P<0.01) and decreased the efflux of EtBr. Furthermore, a time-kill assay revealed that combinations of levofloxacin with ketoconazole or carbonyl cyanide m-chlorophenylhydrazone showed no growth for the tested strains after 24 h in comparison to the effect of levofloxacin alone. Docking studies and the ability of ketoconazole to diminish the relative expression of NorA gene in comparison to control (untreated strains) confirmed its action as an efflux pump inhibitor. Conclusion: The findings showed that the antifungal ketoconazole has no antibacterial activity but can potentiate the activity of the fluroquinolones against MDR S. aureus via inhibiting efflux pump and biofilm formation in vitro.

Introduction: Escherichia (E.) coli can cause intestinal and extra-intestinal infections which ranged from mild to life-threatening infections. The severity of infection is a product of many factors including virulence properties and antimicrobial resistance. Objectives: To determine the antibiotic resistance pattern, the distribution of virulence factors and their association with one another and with some selected resistance genes. Methods: Virulence properties were analyzed phenotypically while antimicrobial susceptibility was tested by Kirby-Bauer agar disc diffusion method. In addition, 64 E. coli isolates were tested for 6 colicin genes, fimH, hlyA, traT, csgA, crl virulence genes and bla_−CTX-M-15, bla_−oxa-2, and bla_−oxa-10 resistance genes by polymerase chain reaction (PCR). Results: Extra-intestinal pathogenic E. coli isolated from urine and blood samples represented a battery of virulence factors and resistance genes with a great ability to produce biofilm. Also, a significant association (P< 0.05) among most of the tested colicin, virulence and resistance genes was observed. The observed associations indicate the importance and contribution of the tested factors in the establishment and the progress of infection especially with Extra-intestinal E. coli (ExPEC) which is considered a great challenging health problem. Conclusion: There is a need for studying how to control these factors to decrease the rate and the severity of infections. The relationship between virulence factors and resistance genes is complex and needs more studies that should be specific for each area.