Differential DNA accessibility to polymerase enables 30-minute phenotypic ß-lactam antibiotic susceptibility testing of carbapenem-resistant Enterobacteriaceae.

The rise in carbapenem-resistant Enterobacteriaceae (CRE) infections has created a global health emergency, underlining the critical need to develop faster diagnostics to treat swiftly and correctly. Although rapid pathogen-identification (ID) tests are being developed, gold-standard antibiotic susceptibility testing (AST) remains unacceptably slow (1-2 d), and innovative approaches for rapid phenotypic ASTs for CREs are urgently needed. Motivated by this need, in this manuscript we tested the hypothesis that upon treatment with ß-lactam antibiotics, susceptible Enterobacteriaceae isolates would become sufficiently permeabilized, making some of their DNA accessible to added polymerase and primers. Further, we hypothesized that this accessible DNA would be detectable directly by isothermal amplification methods that do not fully lyse bacterial cells. We build on these results to develop the polymerase-accessibility AST (pol-aAST), a new phenotypic approach for ß-lactams, the major antibiotic class for gram-negative infections. We test isolates of the 3 causative pathogens of CRE infections using ceftriaxone (CRO), ertapenem (ETP), and meropenem (MEM) and demonstrate agreement with gold-standard AST. Importantly, pol-aAST correctly categorized resistant isolates that are undetectable by current genotypic methods (negative for ß-lactamase genes or lacking predictive genotypes). We also test contrived and clinical urine samples. We show that the pol-aAST can be performed in 30 min sample-to-answer using contrived urine samples and has the potential to be performed directly on clinical urine specimens.

Evaluation of multiplex tandem PCR (MT-PCR) assays for the detection of bacterial resistance genes among Enterobacteriaceae in clinical urines.

Background: Increasing resistance drives empirical use of less potent and previously reserved antibiotics, including for urinary tract infections (UTIs). Molecular profiling, without culture, might better guide early therapy. Objectives: To explore the potential of AusDiagnostics multiplex tandem (MT) PCR UTI assays. Methods: Two MT-PCR assays were developed successively, seeking 8 or 16 resistance genes. Amplification was tracked in real time, with melting temperatures used to confirm product identity. Assays were variously performed on: (i) extracted DNA; (ii) cultured bacteria; (iii) urine spiked with reference strains; and (iv) bacteria harvested from clinical urines. Results were compared with those from sequencing, real-time SybrGreen PCR or phenotypic susceptibility. Results: Performance was similar irrespective of whether DNA, cultures or urines were used, with >90% sensitivity and specificity with respect to common ß-lactamases, dfr genes and aminoglycoside resistance determinants except aadA1/A2/A3, for which carriage correlated poorly with streptomycin resistance. Fluoroquinolone-susceptible and -resistant Escherichia coli (but not other species) were distinguished by the melting temperatures of their gyrA PCR products. The time from urine to results was <3 h. Conclusions: The MT-PCR assays rapidly identified resistance genes from Gram-negative bacteria in urines as well as from cultivated bacteria. Used directly on urines, this assay has the potential to guide early therapy.

Temocillin against Enterobacteriaceae isolates from community-acquired urinary tract infections: low rate of resistance and good accuracy of routine susceptibility testing methods.

Background: Temocillin is an old 'revived' antibiotic that may play an important role in the treatment of febrile urinary tract infection (UTI). Data regarding its activity against current Enterobacteriaceae isolates as well as the performance of routine susceptibility testing methods are, however, scarce. Objectives: To determine the MICs of temocillin for Enterobacteriaceae strains reflecting the current epidemiology and to analyse the accuracy of three commercial methods. Methods: Enterobacteriaceae isolates causing community-acquired UTI were prospectively collected from September 2015 to January 2017 in two French centres. Temocillin MIC was determined by agar dilution (AD) as the reference method and then compared with: (i) susceptibility testing by disc diffusion; (ii) MIC determination by Etest; and (iii) MIC estimation by the Vitek 2 automated system. Results: A total of 762 Enterobacteriaceae were analysed comprising 658 (86.4%) Escherichia coli and 37 (4.9%) ESBL-producing isolates. Susceptibility rate assessed by AD was 99.6% according to the 8 mg/L clinical breakpoint and was significantly lower against the ESBL-producing isolates than the non-ESBL-producing isolates (94.6% versus 99.9%, P < 0.01). The MIC50 and MIC90 for the total set were 3 and 6 mg/L, respectively. According to the 8 mg/L clinical breakpoint, the major error rate was <1% for disc diffusion and Etest, and significantly higher for Vitek 2 (4.3%, P < 0.01), but still low. No very major error was noticed. Conclusions: Temocillin showed a high level of activity against Enterobacteriaceae from community-acquired UTI and good to excellent reliability of routine methods for susceptibility testing in such a setting.

Prevalence of carbapenem resistance and carbapenemase production among Enterobacteriaceae isolated from urine in the UK: results of the UK infection-Carbapenem Resistance Evaluation Surveillance Trial (iCREST-UK).

Objectives: Although carbapenem susceptibility testing has been recommended for all Enterobacteriaceae from clinical specimens, for practical reasons a carbapenem is not included in many primary antibiotic panels for urine specimens. The 'iCREST' study sought carbapenemase-producing Enterobacteriaceae (CPE) in routine urine specimens yielding Gram-negative growth in five diagnostic laboratories in the UK. We sought also to compare locally and centrally determined MICs of meropenem and ceftazidime/avibactam. Methods: Positive growth from up to 2000 urine specimens per laboratory was plated onto chromID® CARBA SMART agar. Suspected CPE colonies were tested locally by Etest for susceptibility to meropenem and ceftazidime/avibactam, and referred to central laboratories for PCR confirmation of CPE status and microbroth MIC determination. Results: Twenty-two suspected CPE were identified from 7504 urine specimens. Ten were confirmed by PCR to have NDM (5), IMP (2), KPC (2) or OXA-48-like (1) carbapenemases. Locally determined ceftazidime/avibactam MICs showed complete categorical agreement with those determined centrally by microbroth methodology. The seven ceftazidime/avibactam-resistant isolates (MICs ≥256 mg/L) had NDM or IMP metallo-carbapenemases. Conclusions: The frequency of confirmed CPE among Gram-negative urinary isolates was low, at 0.13% (10/7504), but CPE were found in urines at all five participating sites and the diversity of carbapenemase genes detected reflected the complex epidemiology of CPE in the UK. These data can inform local policies about the cost-effectiveness and clinical value of testing Gram-negative bacteria from urine specimens routinely against a carbapenem as part of patient management and/or infection prevention and control strategies.

Association between use of different antibiotics and trimethoprim resistance: going beyond the obvious crude association.

Objectives: To evaluate the association between use of different antibiotics and trimethoprim resistance at the population level. Methods: Monthly primary care prescribing data were obtained from NHS Digital. Positive Enterobacteriaceae records from urine samples from patients between April 2014 and January 2016 in England were extracted from PHE's Second Generation Surveillance System (SGSS). Elastic net regularization and generalized boosted regression models were used to evaluate associations between antibiotic prescribing and trimethoprim resistance, both measured at Clinical Commission Group level. Results: In total, 2 487 635 (99%) of 2 513 285 urine Enterobacteriaceae samples from 1 667 839 patients were tested for trimethoprim resistance. Using both elastic net regularization and generalized boosted regression models, geographical variation in trimethoprim resistance among Enterobacteriaceae urinary samples could be partly explained by geographical variation in use of trimethoprim (relative risk = 1.14, 95% CI = 1.02-1.75; relative influence = 4.1) and penicillins with extended spectrum (mainly amoxicillin/ampicillin in England) (relative risk = 1.19, 95% CI = 1.11-1.30; relative influence = 7.4). Nitrofurantoin use was associated with lower trimethoprim resistance levels (relative risk = 0.83, 95% CI = 0.57-0.96; relative influence = 9.2). Conclusions: Use of amoxicillin/ampicillin explained more of the variance in trimethoprim resistance than trimethoprim use, suggesting that co-selection by these antibiotics is an important driver of trimethoprim resistance levels at the population level. Nitrofurantoin use was consistently associated with lower trimethoprim resistance levels, indicating that trimethoprim resistance levels could be lowered if trimethoprim use is replaced by nitrofurantoin.

Progressive increase of resistance in Enterobacteriaceae urinary isolates from kidney transplant recipients over the past decade: narrowing of the therapeutic options.

BACKGROUND: Antibiotic resistance is an emerging phenomenon in kidney transplantation (KT). METHODS: We compared species distribution and antimicrobial susceptibility patterns in 1052 isolates from urine cultures obtained in 2 different cohorts of kidney transplant recipients in a single center (Cohort A: 189 patients undergoing KT between January 2002 and December 2004 [336 isolates]; Cohort B: 115 patients undergoing KT between January 2011 and December 2013 [716 isolates]). RESULTS: Asymptomatic bacteriuria accounted for most of the isolates (86.9% in Cohort A and 92.3% in Cohort B). Klebsiella pneumoniae (9.5% vs. 15.6%), Pseudomonas aeruginosa (1.8% vs. 7.9%), and Enterobacter cloacae (0.6% vs. 3.1%) were significantly more common in Cohort B. The isolation of K. pneumoniae in Cohort B was associated with the occurrence of acute pyelonephritis (9.8% of all K. pneumoniae isolates vs. 2.8% of the remaining uropathogens; P = 0.001). Non-susceptibility rates among Enterobacteriaceae in Cohort B were higher for every class of antibiotics (P ≤ 0.003) with the exception of fosfomycin. Compared to Cohort A, significant increases were seen in isolates from Cohort B for multidrug-resistant (MDR) (43.9% vs. 67.8%, respectively; P = 0.001), extended-spectrum beta-lactamase (ESBL)-producing (6.6% vs. 26.1%; P = 0.001), and carbapenemase-producing Enterobacteriaceae strains (0.0% vs. 5.0%; P = 0.001). Such differences were mostly attributable to K. pneumoniae (as 54.5% and 13.4% of isolates in Cohort B were ESBL-producing and carbapenemase-producing, respectively). MDR isolates were responsible for 69.1% of episodes of symptomatic urinary tract infection in Cohort B. CONCLUSION: The increase in resistance rates among Enterobacteriaceae uropathogens is significant and may have an effect on KT programs.

Asymptomatic bacteriuria among antenatal women in Lagos.

This cross-sectional study was undertaken to determine the prevalence of asymptomatic bacteriuria (ASB), the commonest bacterial isolates and the antibiotic sensitivity pattern among 556 pregnant women in Lagos University Teaching Hospital (LUTH), Nigeria. Women with a bacterial count over 100,000 colony-forming units per millilitre of the same organisms in paired urine samples were considered to have ASB. The prevalence of ASB was 14.6%. Klebsiella was the commonest micro-organism (39.2%) isolated. ASB was significantly associated with marital status, body mass index and parity. There was a significant relationship between urinary nitrites and ASB. The isolated organisms showed remarkable resistance to commonly prescribed antibiotics such as amoxicillin, cloxacillin and trimethoprim but good sensitivity to ofloxacin, gentamycin and ceftazidime. These facts have implications for the management of ASB in pregnancy.

Epidemiology of Carbapenem-Resistant Enterobacteriaceae in 7 US Communities, 2012-2013.

IMPORTANCE: Carbapenem-resistant Enterobacteriaceae (CRE) are increasingly reported worldwide as a cause of infections with high-mortality rates. Assessment of the US epidemiology of CRE is needed to inform national prevention efforts. OBJECTIVE: To determine the population-based CRE incidence and describe the characteristics and resistance mechanism associated with isolates from 7 US geographical areas. DESIGN, SETTING, AND PARTICIPANTS: Population- and laboratory-based active surveillance of CRE conducted among individuals living in 1 of 7 US metropolitan areas in Colorado, Georgia, Maryland, Minnesota, New Mexico, New York, and Oregon. Cases of CRE were defined as carbapenem-nonsusceptible (excluding ertapenem) and extended-spectrum cephalosporin-resistant Escherichia coli, Enterobacter aerogenes, Enterobacter cloacae complex, Klebsiella pneumoniae, or Klebsiella oxytoca that were recovered from sterile-site or urine cultures during 2012-2013. Case records were reviewed and molecular typing for common carbapenemases was performed. EXPOSURES: Demographics, comorbidities, health care exposures, and culture source and location. MAIN OUTCOMES AND MEASURES: Population-based CRE incidence, site-specific standardized incidence ratios (adjusted for age and race), and clinical and microbiological characteristics. RESULTS: Among 599 CRE cases in 481 individuals, 520 (86.8%; 95% CI, 84.1%-89.5%) were isolated from urine and 68 (11.4%; 95% CI, 8.8%-13.9%) from blood. The median age was 66 years (95% CI, 62.1-65.4 years) and 284 (59.0%; 95% CI, 54.6%-63.5%) were female. The overall annual CRE incidence rate per 100<000 population was 2.93 (95% CI, 2.65-3.23). The CRE standardized incidence ratio was significantly higher than predicted for the sites in Georgia (1.65 [95% CI, 1.20-2.25]; P < .001), Maryland (1.44 [95% CI, 1.06-1.96]; P = .001), and New York (1.42 [95% CI, 1.05-1.92]; P = .048), and significantly lower than predicted for the sites in Colorado (0.53 [95% CI, 0.39-0.71]; P < .001), New Mexico (0.41 [95% CI, 0.30-0.55]; P = .01), and Oregon (0.28 [95% CI, 0.21-0.38]; P < .001). Most cases occurred in individuals with prior hospitalizations (399/531 [75.1%; 95% CI, 71.4%-78.8%]) or indwelling devices (382/525 [72.8%; 95% CI, 68.9%-76.6%]); 180 of 322 (55.9%; 95% CI, 50.0%-60.8%) admitted cases resulted in a discharge to a long-term care setting. Death occurred in 51 (9.0%; 95% CI, 6.6%-11.4%) cases, including in 25 of 91 cases (27.5%; 95% CI, 18.1%-36.8%) with CRE isolated from normally sterile sites. Of 188 isolates tested, 90 (47.9%; 95% CI, 40.6%-55.1%) produced a carbapenemase. CONCLUSIONS AND RELEVANCE: In this population- and laboratory-based active surveillance system in 7 states, the incidence of CRE was 2.93 per 100<000 population. Most CRE cases were isolated from a urine source, and were associated with high prevalence of prior hospitalizations or indwelling devices, and discharge to long-term care settings.

Surveillance of antibiotic susceptibility of Enterobacteriaceae isolated from urine samples collected from community patients in a large metropolitan area, 2010-2012.

Antibiotic susceptibilities of large cohorts of Enterobacteriaceae isolated from urine collected in the community are scarce. We report the susceptibilities of Enterobacteriaceae isolated from urine of non-selected community populations in a metropolitan area (Leeds and Bradford, UK) over 2 years. Isolates (n = 6614) were identified as follows: Escherichia coli (n = 5436), Klebsiella spp. (n = 525), Proteus mirabilis (n = 305), and 15 other species (n = 290); 58 isolates were unidentified. Ampicillin resistance was observed in 53% E. coli and 28% P. mirabilis; ≥34% E. coli and P. mirabilis were non-susceptible to trimethoprim compared to 20% Klebsiella spp.; nitrofurantoin resistance was observed in 3% E. coli and 15% Klebsiella spp. The occurrence of extended-spectrum ß-lactamases (ESBL) was low (6%), as was non-susceptibility to carbapenems, cefipime and tigecycline (<2%). Further surveillance is required to monitor this level of resistance and additional clinical studies are needed to understand the impact on the outcome of current empirical prescribing decisions.

[Purple urine bag syndrome: report of one case]. / Síndrome de la bolsa de orina púrpura: un fenómeno inusual y muy llamativo.

Purple urine bag syndrome is an uncommon but particularly striking phenomenon observed in people with urinary catheters and co-existent urinary tract infections. A chemical reaction between plastic and certain bacterial enzymes results in an intense purple urine color. We report a 72 year-old male with a cystostomy. A purple coloration of his urinary drainage bag and tubing was noted in the context of a urinary tract infection caused by Citrobacter freundii.

Prevalence of urinary colonization by extended spectrum-beta-lactamase Enterobacteriaceae among catheterised inpatients in Italian long term care facilities.

BACKGROUND: Long Term Care Facilities (LTCFs) play a key role in guaranteeing care to patients in developed countries. Many patients, mostly elderly, access LTCFs at some time in their lives, and their healthcare pathways often require them to move back and forth between hospital and outpatient settings. These patterns bring about new challenges regarding infection control, especially healthcare associated infections. METHODS: A point prevalence study was conducted in 23 Italian LTCFs, to identify colonization in patients with urinary catheter (>24 hours). Species identification, susceptibility tests and extended spectrum beta lactamase (ESBL) production screenings were performed using Vitek 2 System. Enterobacteria identified by Vitek 2 System as ESBL-producers or suspected AmpC hyperproducers on the basis of cephamycin resistance, were sent to a research laboratory where they underwent a double-disk synergy test. Finally, ESBL-producers were screened for bla resistance genes by PCR assay. RESULTS: 211 patients with catheter were screened, 185 out of 211 patients showed positive samples for the presence of Enterobacteriaceae, 114 of these 185 patients were colonized by extended spectrum cephalosporins resistant microorganisms. We identified a total of 257 Gram negative pathogens, of which 51.8% (133/257) were extended spectrum cephalosporins resistant. 7 out of 133 cephamycin not susceptible strains proved to be AmpC-type beta-lactamases and 125/133 ESBL-producers; 1 was not further characterized. 43 out of 257 (16.7%) E. coli, 37/257 (14.4%) P. mirabilis, 20/257 (7.8%), P. stuartii, 14/257 (5.4%) M. morganii, 7/257 (2.7%), K. pneumoniae, 4/257 (1.6%) C. koseri proved to be overall ESBL-producers by double-disk synergy test. Third and fourth generation cephalosporin resistant P. mirabilis, P. stuartii and M. morganii strains mainly harboured a blaTEM gene (95.9%), while 89.1% of E. coli were positive for the blaCTX-M determinant by PCR and sequencing. Patients with decubitus had a higher risk of colonization by at least one resistant isolate (p < 0.01). Samples of patients undergoing antibiotic therapy and patients with decubitus showed a higher risk (p < 0.05) of colonization by beta-lactam resistant microorganisms. CONCLUSIONS: These data confirm the presence of high percentages of ESBL-positive Enterobacteria in Italian LTCFs and the predominance of CTX-M type ESBL in E. coli. The alarming presence of ESBL-producing Enterobacteriaceae in Italian LTCFs can seriously compromise the effectiveness of antibiotic therapy.acilities (LTCFs), Antimicrobial resistance.

[Evaluation of the ChromID ESBL agar for the detection of ESBL-positive Enterobacteriaceae and vancomycin-resistant enterococcus isolates from urine cultures]. / GSBL Pozitif Enterobacteriaceae ve Vankomisine Dirençli Enterokoklarin Idrar Kültürlerinden Erken Tespitinde ChromID ESBL Agarin Degerlendirilmesi.

Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae strains are frequent causative agents both in community-acquired infections and in nosocomial infections. The newly developed ChromID ESBL agar (bioMerieux, Marcy I'Etoile, France) is a chromogenic medium that helps rapid identification of ESBL-positive Enterobacteriaceae species from the clinical samples. The aim of this study was to evaluate the performance of ChromID ESBL agar in the rapid identification of ESBL-positive pathogens from the urine samples of the patients with urinary tract infections. A total of 672 urine samples (437 outpatients, 235 inpatients) were included in the study. All of the samples were inoculated simultaneously to 5% sheep blood agar, McConkey agar and ChromID ESBL agar media, and evaluated after incubation at 37°C for 18-24 hours. Gram-negative pathogens were tested for ESBL both by the standard combined double-disk diffusion (CDD) method using ceftazidime and cefotaxime disks and by doubledisk synergy (DDS) test. Among 672 urine cultures, 199 yielded microbial growth in routine media (sheep blood agar and/or McConkey agar), whereas 57 yielded bacterial growth in ChromID ESBL agar. When CDD method was accepted as the reference method according to Clinical and Laboratory Standards Institute (CLSI) recommendations, the sensitivity, specificity, positive and negative predictive values for ChromID ESBL agar for the detection of ESBL-positive bacteria in urinary tract infections were estimated as 97%, 92.9%, 89.1%, and 98.1%, respectively. Additionally, we also discovered that Chrom ID ESBL agar could detect vancomycin-resistant enterococci (VRE) as well as ESBL-positive bacteria, in our study. In order to investigate this observation we inoculated a total of 203 stock strains of Enterococcus spp. (118 vancomycin-sensitive, 85 vancomycin-resistant) to this medium. None of the vancomycinsensitive Enterococcus spp. did grow in ChromID ESBL medium, while 83 of the 85 resistant isolates (97.6%) did grow in the medium. As a result, it was concluded that ChromID ESBL agar medium was advantageous since it led to the growth of VRE and ESBL-positive Enterobacteriaceae isolates in different colors and helped in early identification of these two problematic bacteria. We thought that especially early detection of VRE will accelerate the establishment of necessary measures to prevent the nosocomial spread of this microorganism.

Ertapenem for the treatment of urinary tract infections caused by extended-spectrum ß-lactamase-producing bacteria in children.

BACKGROUND: Urinary tract infections (UTIs) are a problem frequently encountered by paediatric healthcare providers. Recent data suggest that extended-spectrum ß-lactamase (ESBL)-producing bacteria are an emerging cause of UTIs in non-hospitalized patients. We report our experience of ertapenem use in 50 patients with complicated UTIs, mainly pyelonephritis, caused by ESBL-producing organisms. METHODS: Fifty patients aged <16 y who had a complicated UTI caused by ESBL-producing organisms and who were treated with ertapenem at our hospital from 1 January 2009 to 31 December 2009, were included in the study. RESULTS: There were 20 (40%) males and 30 (60%) females with a mean ± standard deviation age of 38.6 ± 36.9 months (range 6-156 months). Twenty-eight patients had no urological abnormality. In 40 patients ertapenem was initiated after results of microbiological cultures became available. Ertapenem was initiated empirically for 10 patients known to be colonized with ESBL-producing bacteria. Urine cultures were negative at 3.3 ± 0.7 days (range 2-5 days) after starting ertapenem treatment. The mean duration of ertapenem treatment was 7.8 ± 1.2 days (range 7-14 days). No laboratory or clinical side effects were observed. CONCLUSIONS: Ertapenem is promising for the culture-guided treatment of ESBL-producing Gram-negative complicated UTIs. Well-designed prospective studies are needed to define the role of ertapenem in treating complicated paediatric UTIs, especially upper UTIs.

Detection and genomic characterization of mcr-9 in Enterobacter hormaechei recovered from a pediatric patient in Lebanon.

BACKGROUND: The emergence and spread of mobilized colistin resistance (mcr) genes are a global health concern. OBJECTIVES: In this study, we report the detection and genomic characterization of mcr-9 in a colistin-susceptible Enterobacter hormaechei (EH23) recovered from a pediatric patient in Lebanon. RESULTS: EH23 was susceptible to colistin with a minimal inhibitory concentration (MIC) of 0.25 mg/L. Studying the mcr-9 genetic environment revealed that it was chromosomal and was bracketed by IS903 and IS26. QseCB, a two-component regulatory system, mediating the inducible expression of mcr-9 gene was not detected within the mcr-9 cassette but elsewhere on the genome. EH23 was 99.96% similar based on average nucleotide identity (ANI) to another mcr-negative E. hormaechei OIPH-N069 isolate recovered from Japan. wgSNP-based phylogenetic analysis divided all mcr-9 positive E. hormaechei isolates into five clades (I to V), with isolates from the same ST being clustered together. CONCLUSION: The silent spread of mcr-9, particularly in the globally successful ST-78 Enterobacter lineage, is worrisome and requires close monitoring in humans and animals.

Epidemiology of Carbapenem-resistant Enterobacteriaceae in Egyptian intensive care units using National Healthcare-associated Infections Surveillance Data, 2011-2017.

Objective: To describe the epidemiology of carbapenem-resistant Enterobacteriaceae (CRE) healthcare-associated infections (HAI) in Egyptian hospitals reporting to the national HAI surveillance system. Methods: Design: Descriptive analysis of CRE HAIs and retrospective observational cohort study using national HAI surveillance data. Setting: Egyptian hospitals participating in the HAI surveillance system. The patient population included patients admitted to the intensive care unit (ICU) in participating hospitals. Enterobacteriaceae HAI cases were Klebsiella, Escherichia coli, and Enterobacter isolates from blood, urine, wound or respiratory specimen collected on or after day 3 of ICU admission. CRE HAI cases were those resistant to at least one carbapenem. For CRE HAI cases reported during 2011-2017, a hospital-level and patient-level analysis were conducted using only the first CRE isolate by pathogen and specimen type for each patient. For facility, microbiology, and clinical characteristics, frequencies and means were calculated among CRE HAI cases and compared with carbapenem-susceptible Enterobacteriaceae HAI cases through univariate and multivariate logistic regression using STATA 13. Results: There were 1598 Enterobacteriaceae HAI cases, of which 871 (54.1%) were carbapenem resistant. The multivariate regression analysis demonstrated that carbapenem resistance was associated with specimen type, pathogen, location prior to admission, and length of ICU stay. Between 2011 and 2017, there was an increase in the proportion of Enterobacteriaceae HAI cases due to CRE (p-value = 0.003) and the incidence of CRE HAIs (p-value = 0.09). Conclusions: This analysis demonstrated a high and increasing burden of CRE in Egyptian hospitals, highlighting the importance of enhancing infection prevention and control (IPC) programs and antimicrobial stewardship activities and guiding the implementation of targeted IPC measures to contain CRE in Egyptian ICU's .

Prevalence of non-extended spectrum ß-lactamases SHV-1 and TEM-1 or -2 types in multidrug-resistant Enterobacteriaceae in northern Iran.

The present study aimed to evaluate TEM-1 or -2 and SHV-1 ß-lactamases frequency in multidrug-resistant (MDR) Enterobacteriaceae isolated from patients' urine in northern Iran. The resistance pattern to 20 antibiotics and ESBL production in 200 MDR Enterobacteriaceae was detected using the disk diffusion test and double-disk synergy test (DDST), respectively. Multiplex PCR was applied to detect blaTEM-1 or -2 and blaSHV genes in isolates. DDST findings were inconsistent with multiplex PCR results. The distribution of each of blaTEM-1 or -2 and blaSHV genes, either alone or in combination, in the ESBL-producing isolates was higher than the non-ESBL-producing isolates. There was a significant effect of the presence of blaTEM-1 or -2 gene on resistance to cephalotin at the p < 0.01 level and cefepime, tetracycline, and streptomycin at the P < 0.05 level, and the presence of blaSHV-1 gene on resistance to fosfomycin at the P < 0.05 level as well as the presence both blaTEM-1 or -2 and blaSHV-1 genes on resistance to cephalotin and fosfomycin at the P < 0.01 level. In all isolates, ESBL production, except for cephalotin resistance, did not improve resistance to other antibiotics used and even non-ESBL-producing isolates showed higher resistance to antibiotics compared to ESBL-producing isolates. It seems that mechanisms other than production of ESBL to be involved as part of the resistance mechanisms of the studied isolates against the used antibiotics. For epidemiological studies, both phenotypic and molecular tests must be included to identify the blaTEM-1 or -2 and blaSHV-1 genotypes to ensure infection prevention and control.

Bathroom contamination by antibiotic-resistant Enterobacterales (ESBLPE and CPE): an experimental study.

BACKGROUND: Extended-spectrum ß-lactamase-producing Enterobacterales (ESBLPE) and carbapenemase-producing Enterobacterales (CPE) cause serious infections. Their presence in urine may lead to environmental contamination potentially responsible for cross-transmission. AIM: To evaluate the level of spraying and contamination after emptying urine in the toilet and rinsing in the sink, a common practice in the healthcare setting. METHODS: For each test, the procedure was similar: seat raised, emptying urinal bottle into the toilet at the height of the bowl, rinsing in the sink and flushing. To study splash-drops, water and fluorescein were mixed in the urinal bottle. In each area, the splash-drops frequency and level were assessed with UV. To study contamination, three ESBLPE and one CPE were diluted in saline, 106/mL. Contamination was assessed by sampling before, immediately after and 3 h after the test. The swabs were cultured and the colonies counted and identified. FINDINGS: The areas at the highest risk of spraying were the toilet bowl contour (N = 36/36), the underside of the toilet seat (N = 34) and the inside of the sink (N = 34). Except for gloves (N = 14), there was low clothing contamination. The most frequently contaminated areas were inside the sink (40/48), where the highest levels of contamination were found (14/48). CONCLUSION: Emptying the urinal bottles in the toilet followed by sink rinsing is associated with a significant risk of projection and contamination, depending on the area (highest risk at the sink), but the bacteria did not survive beyond 3 h. This practice, which carries a risk of cross-transmission, should be reviewed.

Prediction of Uropathogens by Flow Cytometry and Dip-stick Test Results of Urine Through Multivariable Logistic Regression Analysis.

PURPOSE: Multidrug-resistant Enterobacteriaceae in urinary tract infection (UTI) has spread worldwide; one cause is overuse of broad-spectrum antimicrobial agents such as fluoroquinolone antibacterials. To improve antimicrobial agent administration, this study aimed to calculate a probability prediction formula to predict the organism strain causing UTI in real time from dip-stick testing and flow cytometry. METHODOLOGY: We examined 372 outpatient spot urine samples with observed pyuria and bacteriuria using dip-stick testing and flow cytometry. We performed multiple logistic-regression analysis on the basis of 11 measurement items and BACT scattergram analysis with age and sex as explanatory variables and each strain as the response variable and calculated a probability prediction formula. RESULTS: The best prediction formula for discrimination of the bacilli group and cocci or polymicrobial group was a model with 5 explanatory variables that included percentage of scattergram dots in an angular area of 0-25° (P<0.001), sex (P<0.001), nitrite (P = 0.002), and ketones (P = 0.133). For a predicted cut-off value of Y = 0.395, sensitivity was 0.867 and specificity was 0.775 (cross-validation group: sensitivity = 0.840, specificity = 0.760). The best prediction formula for P. mirabilis and other bacilli was a model with percentage of scattergram dots in an angular area of 0-20° (P<0.001) and nitrite (P = 0.090). For a predicted cut-off value of Y = 0.064, sensitivity was 0.889 and specificity was 0.788 (cross-validation group: sensitivity = 1.000, specificity = 0.766). CONCLUSION: Simultaneous use of the calculated probability prediction formula with urinalysis results facilitates real-time prediction of organisms causing UTI, thus providing helpful information for empiric therapy.

Prevalence of extended-spectrum beta-lactamase-producing enterobacterial urinary infections and associated risk factors in small children of Garoua, Northern Cameroon.

INTRODUCTION: the emergence of extended-spectrum beta-lactamase-producing Entero bacteriaceae (E-ESBLs) is currently a major public health problem in the world and, in particular, in developing countries. In Cameroon, data on E-ESBLs are rare, especially in Garoua and in the northern region of the country. The objective of this study is to document the epidemiology of E-ESBL infections in small children and to explore their associations with possible risk factors. METHODS: this was a cross-sectional, descriptive study conducted from June 14 to September 30, 2018, including small children with suspected urinary tract infections (UTI) attending the outpatient pediatric departments of two health facilities in the city of Garoua. Urine samples were analyzed at the Bacteriology Laboratory of the Pasteur Center of Cameroon, Annex Garoua. Bacterial culture was carried out on Bio-Rad UriSelect® chromogenic agar and the identification was confirmed by bioMérieux API 20E. The antibiotic susceptibility was determined using the bioMérieux ATB UR gallery and the ESBL phenotype was detected by the double disk synergy method according to the CA-SFM 2013 recommendations. The data was analyzed with the R Statistical Software version 2.15.2. RESULTS: a total of 57 urine samples were collected from children aged from one month to two years, 37 boys and 20 girls. Bacteria were detected by culture in 20 samples: Escherichia coliwas the most frequently (75 %) isolated species followed by Klebsiella pneumoniae(25%). More than half of the infected samples (55%) contained E-ESBL. The presence of an ESBL was significantly associated with previous antibiotic intake up to 3 months prior current UTI (p=0.01664). The E-ESBL strains showed co-resistance to different antibiotics. CONCLUSION: this study reveals the important dissemination of E-ESBLs among small children in the community and a high rate of co-resistance to the different antibiotic families commonly used.