Introduction: Resistance to fluoroquinolones (FQ) in uropathogenic Escherichia coli (UPEC) has emerged as a growing problem. Chromosomal mutations and plasmidmediated quinolone resistance (PMQR) determinants have been implicated. Data concerning the prevalence of these determinants in UPEC in our hospital are quite limited. Purpose: To investigate the occurrence and genetic determinants of FQ resistance in UPEC isolated from urinary tract infection (UTI) cases in Zagazig University Hospitals. Patients and Methods: Following their isolation, the identification and susceptibility of UPEC isolates were performed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometer (MALDI-TOF MS). FQ resistance was detected by the disc diffusion method. Ciprofloxacin minimal inhibitory concentration (MIC) was determined using E-test. Chromosomal mutations in the gyrA gene were detected using polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP), and for detection of PMQR, a couple of multiplex PCR reactions were used. Results: Among a total of 192 UPEC isolates, 46.9% (n=90) were FQ resistant. More than half of the isolates (57.8%) exhibited high-level ciprofloxacin resistance (MIC > 32 µg/mL). Mutations in gyrA were detected in 76.7% of isolates, with 34.4% having mutations at more than one site. PMQR determinants were detected in 80.1% of UPEC isolates, with aac(6ʹ)-Ibcr gene being the most frequent found in 61.1% of isolates. Conclusion: There is a high prevalence of both gyrA mutations and PMQR determinants among UPEC isolates in our hospital which contribute to high-level ciprofloxacin resistance, a finding that may require the revision of the antibiotics used for empirical treatment of UTI.

Background. Dysbiosis of gut microbiota could promote autoimmune disorders including systemic lupus erythematosus (SLE). Clarifying this point would be of great importance in understanding the pathogenesis and hence the development of new strategies for SLE treatment. Aim. This study aimed to determine the fecal microbiota profile in newly diagnosed SLE patients compared to healthy subjects and to investigate the correlation of this profile with disease activity. Methods. Newly diagnosed SLE patients who fulfilled at least four of the American College of Rheumatology (ACR) criteria were enrolled during the study period. Patients with lupus were matched to healthy subjects. SLE activity was evaluated using the Systemic Lupus Disease Activity Index (SLEDAI-2K). Fresh fecal samples were collected from each subject. Genomic DNA was extracted from fecal samples. Quantitative real-time PCR was applied for quantitation of Firmicutes phylum, Bacteroidetes phylum, and Lactobacillus genus in comparison to the total fecal microbiota. Results of patients’ samples were compared to those of healthy subjects and were correlated to patients’ SLEDAI-2K score. Results. Twenty SLE patients’ samples were compared with 20 control samples. There was a significant alteration in SLE patients’ gut microbiota. A significantly lower ( p ≤ 0.001 ) Firmicutes/Bacteroidetes (F/B) ratio in SLE patients (mean ratio: 0.66%) compared to healthy subjects (mean ratio: 1.79%) was found. Lactobacillus showed a significant decrease in SLE patients ( p = 0.006 ) in comparison to healthy controls. An inverse significant correlation between SLEDAI-2K scores for disease activity and F/B ratio (r = −0.451; p = 0.04 ) was found. However, an inverse nonsignificant correlation between SLEDAI-2K scores for disease activity and Lactobacillus (r = −0.155; p = 0.51 ) was detected. Conclusion. Compared to healthy controls, recently diagnosed SLE Egyptian patients have an altered fecal microbiota profile with significant lowering of both F/B ratio and Lactobacillus abundance, which is weakly correlated with disease activity.

Objective: The coexistence of ESBLs and pAmpCs enzymes in the same Klebsiella strain may result in false-negativetests for the detection of ESBLs as pAmpCs resist inhibition by clavulanic acid so masking ESBL presence. This study wasto highlight the detection rates of ESBLs and pAmpCs by using phenotypic method; MAST 4-disc test and multiplexpolymerase chain reaction (PCR) method. In addition, it aimed to evaluate the sensitivity of the phenotypic method indetection of these enzymes.Methods: Klebsiella isolates were collected from clinical samples in different wards in Zagazig University Hospitals. Theantibiogram of these bacteria was determined by disc diffusion method. The presence of ESBLs and pAmpCs within theisolates was determined using the phenotypic MAST 4-disc test followed by a multiplex PCR method.Results: In total, 38 Klebsiella pneumoniae strains were evaluated. Among these isolates, 65.8% were ESBL producers,2.6% were pAmpC producers, and 31.6% were neither ESBL nor pAmpC producers. The most frequent genotype of ESBLwas CMY (84%); followed by CMY (44%) before pAmpC producers were of CMY genotype. The distribution of differentESBL genotypes was CMY, CMY and CTX-M II genotype (28%) and followed by CMY and CTX-M IV genotype (24%). Usingmultiplex PCR as a reference method, MAST 4-disc test was of 92% sensitivity and 86.7% specificity.Conclusion: A rising alarm of ESBL producing strains among K. pneumoniae isolates. The exact detection of ESBLs inisolates that produce both enzymes is important for both treatment and epidemiology. J Microbiol Infect Dis 2013; 3(1):24-30Key words: ESBL, pAmpC, Β-Lactasmases, Nosocomial, Klebsiell

Background: Klebsiella pneumoniae (K. pneumoniae) significantly contributes to hospital-acquired bloodstream infections with high morbidity and mortality, especially biofilm-producing strains. It has been noticed that ability to produce biofilm by K. pneumoniae is linked to presence of some virulence genes, but this relationship needs further investigation. This work aimed to investigate ten virulence genes that may contribute to biofilm formation in K. pneumoniae strains causing bloodstream infections. Methods: This cross-sectional study included 108 K. pneumoniae isolates obtained from cases of hospital-acquired bloodstream Infections. The sensitivity to different antibiotics was tested by the disc diffusion method. Ability to form biofilm was detected by the tissue culture plate method. All isolates were tested by polymerase chain reaction (PCR) to detect ten virulence genes suggested to be linked to biofilm formation ability. Results: The ability of biofilm formation was detected in 55.6% of the studied strains. Biofilm formation was more prevalent among wcaG, fimH, wabG, and mrkD positive isolates in comparison to negative isolates for the same genes. However, only wcaG and fimH genes have been found to be significantly associated with biofilm formation (P < 0.05). Conclusion: The association between the ability to form biofilm and the existence of wcaG and fimH genes in K. pneumoniae bacteremia isolates suggests these genes as promising therapeutic targets.