Emergence of Multidrug-resistant Carbapenemases and MCR-1 Producing Pseudomonas aeruginosa in Egypt

Pseudomonas aeruginosa is an expedient Gram-negative bacterium, which is characterized by its ability to acquire antimicrobial resistance. In this study, 56 unrepeatable carbapenem-resistant P. aeruginosa isolates were gathered from various clinical sources from hospitals in Cairo and Mansoura universities. The isolates exhibited diminished susceptibility towards carbapenems, quinolones, aminoglycosides and chloramphenicol by using disc diffusion method. Carbapenemase production was confirmed among the isolates, where all the 56 P. aeruginosa isolates harboured carbapenemase genes including blaVIM (43 isolates), blaKPC (38 isolates), blaNDM-1 (17 isolates), blaIMP (16 isolates) and blaOXA-48 (15 isolates). Among the isolates, 13 carried only one carbapenemase gene, while 43 isolates carried multiple carbapenemase genes. MCR-1 production was confirmed in 10 of the tested isolates by detecting the mcr-1 gene encoding for the colistin resistance. Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) evaluation showed that the tested isolates were unrelated to each other. Therefore, this study rises the danger of emergence of MDR P. aeruginosa resistant to carbapenems coupled with other antimicrobials including colistin, which is regarded as the last reservoir for the management of infections caused by MDR Gram-negative pathogens. Early inspection of resistance patterns in MDR organisms is an important tool to control and prevent infections via limiting the spread of these pathogens

In-vitro investigation of biofilm-specific resistance and virulence of biofilm-forming uropathogenic Escherichia coli

Uropathogenic Escherichia coli (UPEC) is the primary etiologic agent of urinary tract infections (UTIs). This study aimed to investigate the difference in antimicrobial susceptibility of UPEC isolates in the planktonic and biofilm states. Important virulence factors were also evaluated. The minimum inhibitory concentrations (MICs) were determined and recorded as 0.5-64 μg/ ml for amikacin, 0.5-64 μg/ ml for cefotaxime, 0.25-64 μg/ ml for cefepime, 0.25-16 μg/ ml for meropenem, and 0.125-32 μg/ ml for ciprofloxacin. Biofilm-specific resistance was assessed using the minimum biofilm eradication concentration (MBEC). The obtained results for MBEC were: 8-512 μg/ ml for amikacin, 32-512 μg/ ml for cefotaxime, 8-512 μg/ ml for cefepime, 4-256 μg/ ml for meropenem, and 4-128 μg/ ml for ciprofloxacin. The virulence factors were evaluated using suitable phenotypic techniques. Our findings revealed a significant enhancement in the antimicrobial resistance after biofilm formation. The MBEC values were higher than the MIC values by 2-128 folds for amikacin, 2-256 folds for cefotaxime, 2-64 folds for cefepime, 8-128 folds for meropenem, and 4-128 folds for ciprofloxacin. The swimming and swarming motilities demonstrated a significant positive correlation (rs = 0.506, P< 0.001). Protease production analysis revealed a large variation, with the weak biofilm-producing isolates EW2 and EW15 displaying the largest zone diameters of 39 mm and 33 mm; respectively. We have also evaluated the distribution and levels of siderophore production, which were significantly associated with meropenem resistance. Finally, this study underscores the importance of considering biofilm formation in UPEC treatment and emphasizes the need for therapeutics targeting these biofilms

Co-existence of NDM-1 and OXA-48 genes in Carbapenem Resistant Klebsiella pneumoniae clinical isolates in Kafrelsheikh, Egypt

Background: The noteworthy spread of carbapenem-resistant K. pneumoniae (CR-KP) isolates represents a significant safety threat. Objective: Determination of the carbapenemase genes incidence among CR-KP clinical isolates in Kafrelsheikh, Egypt. Methods: A total of 230 K. pneumoniae isolates were recovered from four hospitals in Kafrelsheikh, Egypt. Susceptibility testing was conducted using Kirby-Bauer method and automated-Vitek2 system. CR-KP isolates were tested using modified Hodge test (MHT) and combined disk synergy test. PCR and DNA sequencing were conducted for CR-KP isolates to recognize the included carbapenemase-genes. Results: Out of 230 K. pneumoniae isolates, 50 isolates presented resistance to carbapenem (meropenem). All 50 CR-KP isolates were multidrug-resistant (MDR). Genes like blaNDM-1 and blaOXA-48 were the only detected genes among CR-KP with an incidence of 70.0% and 52.0%, respectively. Up to 74.0% of the tested isolates carried at least one of the two recorded genes, among them 48.0% co-harbored both blaNDM-1 and blaOXA-48 genes. The accession-numbers of sequenced blaNDM-1 and blaOXA-48 genes were MG594615 and MG594616, respectively. Conclusion: This study reported a high incidence of MDR profile with the emergence of blaNDM-1 and blaOXA-48 genes co-existence in CR-KP isolates in Kafrelsheikh, Egypt. Hence, more restrictions should be applied against the spread of such serious pathogens

Effect of carbapenem resistance on outcomes of bloodstream infection caused by Enterobacteriaceae in low-income and middle-income countries (PANORAMA): a multinational prospective cohort study

Background Low-income and middle-income countries (LMICs) are under-represented in reports on the burden of antimicrobial resistance. We aimed to quantify the clinical effect of carbapenem resistance on mortality and length of hospital stay among inpatients in LMICs with a bloodstream infection due to Enterobacteriaceae. Methods The PANORAMA study was a multinational prospective cohort study at tertiary hospitals in Bangladesh, Colombia, Egypt, Ghana, India, Lebanon, Nepal, Nigeria, Pakistan, and Vietnam, recruiting consecutively diagnosed patients with carbapenem-susceptible Enterobacteriaceae (CSE) and carbapenem-resistant Entero-bacteriaceae (CRE) bloodstream infections. We excluded patients who had previously been enrolled in the study and those not treated with curative intent at the time of bloodstream infection onset. There were no age restrictions. Central laboratories in India and the UK did confirmatory testing and molecular characterisation, including strain typing. We applied proportional subdistribution hazard models with inverse probability weighting to estimate the effect of carbapenem resistance on probability of discharge alive and in-hospital death, and multistate modelling for excess length of stay in hospital. All patients were included in the analysis. Findings Between Aug 1, 2014, and June 30, 2015, we recruited 297 patients from 16 sites in ten countries: 174 with CSE bloodstream infection and 123 with CRE bloodstream infection. Median age was 46 years (IQR 15–61). Crude mortality was 20% (35 of 174 patients) for patients with CSE bloodstream infection and 35% (43 of 123 patients) for patients with CRE bloodstream infection. Carbapenem resistance was associated with an increased length of hospital stay (3·7 days, 95% CI 0·3–6·9), increased probability of in-hospital mortality (adjusted subdistribution hazard ratio 1·75, 95% CI 1·04–2·94), and decreased probability of discharge alive (0·61, 0·45–0·83). Multilocus sequence typing showed various clades, with marginal overlap between strains in the CRE and CSE clades. Interpretation Carbapenem resistance is associated with increased length of hospital stay and mortality in patients with bloodstream infections in LMICs. These data will inform global estimates of the burden of antimicrobial resistance and reinforce the need for better strategies to prevent, diagnose, and treat CRE infections in LMICs

Panton-Valentine Leukocidin (PVL) genes may not be a reliable marker for community-acquired MRSA in the Dakahlia Governorate, Egypt

Abstract Background Methicillin-resistant Staphylococcus aureus is linked to both nosocomial and community infections. One of the key virulence factors of S. aureus is Panton-Valentine leukocidin (PVL). The PVL genes are mostly associated with community-acquired MRSA (CA-MRSA). This study evaluates the prevalence of PVL genes as a marker for CA-MRSA at tertiary hospitals in Mansoura, Dakahlia, Egypt. S. aureus was isolated from clinical specimens obtained from different departments of tertiary hospitals, outpatient clinics, and hospital healthcare workers (HCWs). PCR was used to detect the mecA, PVL, and SCCmec genes among the recovered isolates. Standard broth microdilution method was used to determine the minimum inhibitory concentrations (MIC) of nine antibiotics against S. aureus. Results Two hundred S. aureus isolates were recovered and identified out of the total isolates (n = 320). The mecA gene was detected in 103 S. aureus isolates (51.5%). Among the MRSA isolates, 46.60% were PVL-positive. The incidence of the PVL genes of MRSA in nosocomial (HA), outpatient clinics (CA), and HCWs was 46.66%, 56.52%, and 42%, respectively. All MRSA isolates showed resistance to cefoxitin. The percentage of resistance to most tested antibiotics was high, except for ciprofloxacin (6.85%). Both antibiotic resistance and multidrug resistance among MRSA isolates were generally higher in PVL-positive isolates than in PVL-negative isolates in HA- and CA-MRSA isolates. While SCCmec type V was the most prevalent in PVL-positive MRSA stains, type I was the most prevalent in PVL-negative isolates. Conclusion This study revealed that PVL genes are generally highly prevalent among mecA-positive MRSA isolates, whether they are CA-MRSA, HA-MRSA, or HCW isolates. Therefore, PVL is not a valid marker for CA-MRSA in Mansoura, Dakahlia Governorate, Egypt, as has been reported in other countries. Further epidemiologic studies are required to track the incidence of PVL in HA-MRSA, CA-MRSA, and HCW isolates in other Egyptian governorates

Description of the Gram-Negative, Obligately Aerobic, Nitrilotriacetate (NTA)-Utilizing Bacteria as Chelatobacter heintzii, gen. nov., sp. nov., and Chelatococcus asaccharovorans, gen. nov., sp. nov

Nine obligately aerobic, Gram-negative, nitrilotriacetate (NTA)-utilizing isolates and the two NTA-utilizing reference strains Pseudomonas spp. ATCC 29600 and ATCC 27109 were investigated by a polyphasic taxonomic approach. The presence of sym-homospermidine as the major polyamine excluded all NTA-utilizing bacteria from the authentic genus Pseudomonas and suggested their allocation into the alpha-2 subclass of the Proteobacteria. Protein patterns and DNA hybridization studies revealed two unrelated groups, neither of which could be allocated to any currently recognized genus. The 16S rRNA fragment sequences (position 1220–1377, Escherichia coli-nomenclature) of representative strains from both groups differ by 13 nucleotides. This indicates that they are not closely related, and justifies placement of the NTA-utilizing bacteria in two different genera, Chelatobacter, gen. nov., and Chelatococcus, gen. nov. These taxa are formally described below. A difference of 10 nucleotides indicated that Rhodopseudomonas acidophila is the most closely related neighbour of Chelatococcus asaccharovorans TE2. Oligonucleotide cataloguing places Chelatobacter heintzii ATCC 29600 next to the carbon monoxide oxidizing bacterium “Alcaligenes carboxydus” DSM 1086, within the Agrobacterium-Rhizobium branch

Efficiency of xylanase, emulsifier, and guanidinoacetic acid in restoring energy deficit in male broilers fed low metabolisable energy diets

The study aimed to examine the distinct impacts of incorporating xylanase (Xyl), emulsifier (EM), and guanidinoacetic acid (GAA) as dietary supplements in low metabolisable energy (ME) diets on performance, energy and protein efficiency ratios, oxidative biomarkers, gene expression, and gut morphology. Seven hundred one-day-old (Ross 308) male-broilers were assigned to 5 dietary treatments with 5 replicates of 28 birds each. The experimental group denoted as the positive control (PC) fed on diets in accordance with the breed recommendations. The negative control (NC) was subjected to a dietary intervention reducing the ME by 200 kcal/kg compared to the PC. The remaining experimental diets comprised NC diet that were supplemented with 0.01% Xyl, 0.03% EM and 0.06% GAA. Results showed that birds fed low-ME-diets increased their voluntary feed intake to meet their energy needs but was at the expense of their productive efficiency. Only NC+GAA partially restored broiler performance compared to PC. However, compared to the PC group Xyl, EM and GAA improved the energy and growth-related gene expression, oxidative biomarkers, and gut histomorphology (p &lt; 0.05). The key features associated with Xyl and EM were growth-related genes and intestinal mucus, while GAA was associated with energy-related genes, oxidative biomarkers and jejunum-villi height and villus: crypt ratio. In conclusion, Xyl, EM and GAA supplementation to NC group were able to improve the health status of the birds. However, to improve the production efficiency, future research is needed to elucidate the effect of combined products in birds fed on such low ME diets. &nbsp

Additional file 1 of Genetic diversity of macrolides resistant Staphylococcus aureus clinical isolates and the potential synergistic effect of vitamins, C and K3

Additional file 1

Serum calprotectin as a potential biomarker for subclinical enthesitis in psoriatic patients

Aim of the work: To assess serum calprotectin level in psoriatic patients and investigate its potential relation to clinical and ultrasonographic enthesitis. Patients and methods: The study included 45 psoriatic patients and 20 matched healthy controls. Enthesitis was assessed clinically, by musculoskeletal ultrasound (MSUS) and power Doppler, by Leeds Enthesitis Index (LEI), Maastricht Ankylosing Spondylitis Enthesitis (MASES), Spondyloarthritis Research Consortium of Canada (SPARCC) and Madrid Sonography Enthesitis Index (MASEI) scores. The Psoriasis Area, Severity Index (PASI) and Dermatology Life Quality Index (DLQI) were evaluated. Serum calprotectin was measured using enzyme-linked immunosorbent assay. Results: The study included 45 psoriatic patients with a mean age of 49.9 ± 7.8 years and they were 23 males and 22 females with disease duration of 5.2 ± 3 years, (0.5–11 years). Patients were categorized into those with enthesitis (n = 25; 20 clinically and 5 subclinically diagnosed by MSUS) and those without (n = 20). Serum calprotectin was significantly higher among patients with enthesitis (clinical 593.7 ± 192.5 ng/ml and subclinical 692 ± 265.9 ng/ml) compared to those without (381.2 ± 198.5 ng/ml) and to the control (111.1 ± 15 ng/ml) (p = 0.001). The DLQI was significantly severer in patients with clinical enthesitis compared to those subclinically detected or without (p < 0.001). The acute phase reactants (erythrocyte sedimentation rate and C-reactive protein) were comparable between patients and control. Serum calprotectin significantly correlated with MASES, LEI, SPARCC and MASEI score in psoriatic patients (p < 0.001). At cutoff 141 ng/ml, calprotectin yielded specificity 69% and sensitivity 75% to detect enthesitis (p = 0.008). Conclusion: Calprotectin may be considered as a potential biomarker for detection of enthesitis in psoriatic patients