Proteus mirabilis is a Gram negative bacterium that is a frequent cause of catheter-associated urinary tract infections (CAUTIs). Its ability to cause such infections is mostly related to the formation of biofilms on catheter surfaces. In order to form biofilms, P. mirabilis expresses a number of virulence factors. Such factors may include adhesion proteins, quorum sensing molecules, lipopolysaccharides, efflux pumps, and urease enzyme. A unique feature of P. mirabilis biofilms that build up on catheter surfaces is their crystalline nature owing to their ureolytic biomineralization. This leads to catheter encrustation and blockage and, in most cases, is accompanied by urine retention and ascending UTIs. Bacteria embedded in crystalline biofilms become highly resistant to conventional antimicrobials as well as the immune system. Being refractory to antimicrobial treatment, alternative approaches for eradicating P. mirabilis biofilms have been sought by many studies. The current review focuses on the mechanism by which P. mirabilis biofilms are formed, and a state of the art update on preventing biofilm formation and reduction of mature biofilms. These treatment approaches include natural, and synthetic compounds targeting virulence factors and quorum sensing, beside other strategies that include carrier-mediated diffusion of antimicrobials into biofilm matrix. Bacteriophage therapy has also shown successful results in vitro for combating P. mirabilis biofilms either merely through their lytic effect or by acting as facilitators for antimicrobials diffusion.

Fluoroquinolones have been used for prophylaxis against infections in cancer patients but their impact on the resistance mechanisms still require further investigation. To elucidate mechanisms underlying ciprofloxacin (CIP) resistance in Gram-negative pathogens causing infections to cancer patients, 169 isolates were investigated. Broth microdilution assays showed high-level CIP resistance in 89.3% of the isolates. Target site mutations were analyzed using PCR and DNA sequencing in 15 selected isolates. Of them, all had gyrA mutations (codons 83 and 87) with parC mutations (codons 80 and 84) in 93.3%. All isolates were screened for plasmid-mediated quinolone resistance (PMQR) genes and 56.8% of them were positive in this respect. Among PMQR genes, aac(6′)-Ib-cr predominated (42.6%) while qnr genes were harbored by 32.5%. This comprised qnrS in 26.6% and qnrB in 6.5%. Clonality of the qnr-positive isolates using ERIC-PCR revealed that most of them were not clonal. CIP MIC reduction by CCCP, an efflux pump inhibitor, was studied and the results revealed that contribution of efflux activity was observed in 18.3% of the isolates. Furthermore, most fluoroquinolone resistance mechanisms were detected among Gram-negative isolates recovered from cancer patients. Target site mutations had the highest impact on CIP resistance as compared to PMQRs and efflux activity.

Acinetobacter baumannii has become a major challenge to clinicians worldwide due to its high epidemic potential and acquisition of antimicrobial resistance. This work aimed at investigating antimicrobial resistance determinants and their context in four extensively drug-resistant (XDR) NDM-producing A. baumannii clinical isolates collected between July and October 2020 from Kasr Al-Ainy Hospital, Cairo, Egypt. A total of 20 A. baumannii were collected and screened for acquired carbapenemases (bla_NDM, bla_VIM and bla_IMP) using PCR. Four NDM producer A. baumannii isolates were identified and selected for whole-genome sequencing, in silico multilocus sequence typing, and resistome analysis. Antimicrobial susceptibility profiles were determined using disk diffusion and broth microdilution tests. All bla_NDM-positive A. baumannii isolates were XDR. Three isolates belonged to high-risk international clones (IC), namely, IC2 corresponding to ST570^Pas/1701^Oxf (M20) and IC9 corresponding to ST85^Pas/ST1089^Oxf (M02 and M11). For the first time, we report bla_NDM-1 gene on the chromosome of an A. baumannii strain that belongs to sequence type ST164^Pas/ST1418^Oxf. Together with AphA6, bla_NDM-1 was bracketed by two copies of ISAba14 in ST85^Pas isolates possibly facilitating co-transfer of amikacin and carbapenem resistance. A novel bla_ADC allele (bla_ADC-257) with an upstream ISAba1 element was identified in M19 (ST/CC164^Pas and ST1418^Oxf/CC234^Oxf). bla_ADC genes harbored by M02 and M11 were uniquely interrupted by IS1008. Tn2006-associated bla_OXA-23 was carried by M20. bla_OXA-94 genes were preceded by ISAba1 element in M02 and M11. AbGRI3 was carried by M20 hosting the resistance genes aph(3`)-Ia, aac(6`)-Ib`, catB8, ant(3``)-Ia, sul1, armA, msr(E), and mph(E). Nonsynonymous mutations were identified in the quinolone resistance determining regions (gyrA and parC) of all isolates. Resistance to colistin in M19 was accompanied by missense mutations in lpxACD and pmrABC genes. The current study provided an insight into the genomic background of XDR phenotype in A. baumannii recovered from patients in Egypt. WGS revealed strong association between resistance genes and diverse mobile genetic elements with novel insertion sites and genetic organizations.

Fluoroquinolones (FQs) are the drugs of choice for prophylaxis of bacterial infections in immunocompromised cancer patients. This study aimed to investigate FQ resistance and the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants in 239 Gram-negative isolates collected at a tertiary care cancer hospital in Cairo, Egypt. Disc diffusion and broth microdilution tests showed that 70.7% of the isolates were nonsusceptible to ciprofloxacin (MIC ₅₀ = 64 μg/ml). Polymerase chain reaction (PCR) revealed that 53.6% of the isolates carried at least one PMQR determinant, of which 23.4% were susceptible to ciprofloxacin. The most prevalent gene, aac(6′)-Ib-cr , was identified in 36.8% of the isolates, while qnr genes were harbored by 31.0% ( qnrS , 24.3%; qnrB , 7.1%, and qnrA , 0.4%). The oqxAB genes were only detected in Klebsiella sp. isolates (92.5%). PMQR determinants were more likely detectable among isolates recovered from pediatric patients than adults (59.3% vs. 43.8%) and were significantly associated with ceftriaxone and gentamicin resistance. A combined genetic analysis using random amplified polymorphic DNA-PCR and enterobacterial repetitive intergenic consensus-PCR showed that most of the qnr -positive isolates were not clonal. Findings of the current study raised concerns about the efficacy of prophylactic use of FQs in cancer patients in our region. It also demonstrates the possible role of PMQR-positive ciprofloxacin-susceptible isolates in the dissemination of resistance to other antimicrobial agents and the urgent need to reconsider the existing FQ breakpoints defined by the Clinical and Laboratory Standards Institute.

Acinetobacter baumannii (A. baumannii) is a highly problematic pathogen with an enormous capacity to acquire or upregulate antibiotic drug resistance determinants. The genomic epidemiology and resistome structure of 46 A. baumannii clinical isolates were studied using whole-genome sequencing. The isolates were chosen based on reduced susceptibility to at least three classes of antimicrobial compounds and were initially identified using MALDI-TOF/MS, followed by polymerase chain reaction amplification of bla_OXA-51-like genes. The susceptibility profiles were determined using a broth microdilution assay. Multi-, extensive-, and pan-drug resistance was shown by 34.8%, 63.0%, and 2.2% of the isolates, respectively. These were most susceptible to colistin (95.7%), amikacin, and trimethoprim/sulfamethoxazole (32.6% each), while only 26.1% of isolates were susceptible to tigecycline. In silico multi-locus sequence typing revealed 8 Pasteur and 22 Oxford sequence types (STs) including four novel STs (ST^Oxf 2805, 2806, 2807, and 2808). The majority of the isolates belonged to Global Clone (GC) 2 (76.4%), GC5 (19.6%), GC4 (6.5%), GC9 (4.3%), and GC7 (2.2%) lineages. An extensive resistome potentially conferring resistance to the majority of the tested antimicrobials was identified in silico. Of all known carbapenem resistance genes, bla_OXA-23 was carried by most of the isolates (69.6%), followed by ISAba1-amplified bla_ADC (56.5%), bla_NDM-1 and bla_GES-11 (21.7% each), and bla_GES-35 (2.2%) genes. A significant correlation was found between carbapenem resistance and carO mutations, which were evident in 35 (76.0%) isolates. A lower proportion of carbapenem resistance was noted for strains possessing both bla_OXA-23- and bla_GES-11. Amikacin resistance was most probably mediated by armA, aac(6′)-Ib9, and aph(3′)-VI, most commonly coexisting in GC2 isolates. No mutations were found in pmrABC or lpxACD operons in the colistin-resistant isolates. Tigecycline resistance was associated with adeS (N268Y) and baeS (A436T) mutations. While the lineage-specific distribution of some genes (e.g., bla_ADC and bla_OXA-51-like alleles) was evident, some resistance genes, such as bla_OXA-23 and sul1, were found in all GCs. The data generated here highlight the contribution of five GCs in A. baumannii infections in Egypt and enable the comprehensive analysis of GC-specific resistomes, thus revealing the dissemination of the carbapenem resistance gene bla_OXA-23 in isolates encompassing all GCs.

Aims: To determine resistance rates and patterns of certain uropathogens, including E. coli, Klebsiella spp. and Pseudomonas spp., isolated from hospitalized urinary tract infections patients, to aminoglycoside antibiotics and to detect the most prevalent plasmid-mediated aminoglycoside modifying enzymes (AMEs). Methods: Uropathogenic isolates (150) were recovered from urine specimens of hospitalized UTI patients in Cairo, Egypt and identified by conventional methods. The recovered uropathogens (E. coli, Klebsiella spp. and Pseudomonas spp.) were tested for their susceptibility to gentamicin, tobramycin, amikacin, neomycin, netilmicin, and kanamycin by disc diffusion method. Plasmid-mediated aminoglycoside resistance was determined by transformation experiments as well as by using plasmids as templates for PCR screening of the AMEs-coding genes aph(3')-I, aac(6')-I, aac(3)-I, aac(3)-II and ant(2'')-I in all resistant isolates. Results: Of a total of 150 uropathogenic clinical isolates, 110 isolates were of the above mentioned genera and were selected for the current study. Sixty three isolates (57.2%) were resistant to at least one aminoglycoside antibiotic. Highest and lowest resistance rates were observed to kanamycin (53.6%) and amikacin (7.2%), respectively. The resistance rates to gentamicin, neomycin, tobramycin and netilmicin were 33.6%, 24.5%, 23.6% and 14.5%, respectively. AMEs-coding genes were detected on the plasmids of 93.6% of resistant isolates with prevalence rates of 53.9% for ant(2'')-I, 38% for both aac(6')-I and aac(3)-II and 33.3% for aph(3')-I, while aac(3)-I gene was not detected in any of the tested resistant isolates. Double and triple combinations of AMEs-coding genes were detected in ich49.2% of resistant isolates. Conclusion: A high prevalence of plasmid-mediated resistance to aminoglycoside antibiotics in Gram negative uropathogens from hospitalized patients was observed. Uropathogens may represent potential reservoirs of panaminoglycoside resistance in hospitals, having on their plasmids combinations of AMEs-coding genes. Good infection control measures in Egyptian hospitals together with periodic screening of prevalence rates of different resistance genes are required.