BACKGOUND The members of the so-called ESKAPE group (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) are a frequent cause of severe infection, ranking among the most relevant causes of hospital infections. In Peru, few studies, often focused in a single ESKAPE microorganism, have been performed, but none providing an overall and comprehensive long-time analysis of the antibiotic resistance of ESKAPE microorganisms. In the present study, the evolution of antimicrobial resistance levels of ESKAPE microorganisms isolated during 2009 - 2010 (Period 1) and 2012 - 2014 (Period 2) in a IV-level hospital in Lima was analyzed. MATERIALS AND METHODS ESKAPE microorganisms were isolated from inpatients clinical samples. Bacterial identification, as well as antimicrobial susceptibility levels for up to 29 antimicrobial agents and presence of Extended-Spectrum β-Lactamases (only established in K. pneumoniae) were determined using automatic methods. RESULTS Of 9,918 clinical isolates, 1,917/3,777 (50.8%) [JAN/2009-JUN/2010 (Period 1)] and 4764/6141 (46.4%) [JAN/2012-DEC/2014 (Period 2)] belonged to the ESKAPE group (P <0.0001). ESKAPE were more frequent in the intensive care unit (ICU) (P <0.0001). E. faecium decreased from 5.1% to 4.1% (P <0.5), S. aureus from 10.5% to 7.0% (P <0.05), and P. aeruginosa from 12.9% to 11.6% (P <0.05), while, A. baumannii increased from 5.0% to 6.7% (P <0.05), mainly related to an increase in ICU isolates (8.4% vs. 17.1%; P <0.05). Overall, high levels of antimicrobial resistance were detected, but with few exceptions (e.g. vancomycin in E. faecium), antibiotic resistance levels remained stable or lower in Period 2. Contrarily, A. baumannii showed significantly increased resistance to different cephalosporins, carbapenems and amoxicillin plus sulbactam. CONCLUSION The introduction of a successful extensively drug-resistant A. baumannii clone in the ICU is suspected. The isolation of ESKAPE and levels of antibiotic resistance levels have reduced over time.

/pdf/evolution-of-antimicrobial-resistance-levels-of-eskape-16lkk23o3c.pdf

Evolution of Antimicrobial Resistance Levels of ESKAPE Microorganisms in a Peruvian IV-Level Hospital

The emergence and transmission of metallo-β-lactamases are significant public health concerns that threaten the utility of antimicrobial therapy (1, 2).….

Characterization of the First Carbapenem-Resistant Pseudomonas aeruginosa Clinical Isolate Harboring blaSIM-1 from the United States.

Background: The opportunistic human pathogen, Pseudomonas aeruginosa, is a critical cause of nosocomial infection with high morbidity and mortality rate. Eradication of P. aeruginosa has been troublesome due to its high capacity to develop strong multidrug resistance (MDR). Objectives: The purposes of this study were to define the pattern of antimicrobial sensitivity, typing, and prevalence of metallo-β-lactamase (MBL) and detect the oprD, blaCTX-M, blaSHV, blaTEM, blaIMP, blaNDM, and blaVIM among clinical isolates of P. aeruginosa collected from Tehran hospitals. Methods: Clinical isolates were collected from hospitalized children in selected hospitals in Tehran from March 2019 to February 2020. The antimicrobial susceptibility test (AST) was performed by the Kirby-Bauer disk diffusion method. Composed disc diffusion tests were performed to screen MBL production. MBLs and extended-spectrum β-lactamases (ESBL) encoding genes were amplified by polymerase chain reaction (PCR). Amplification of the oprD gene were performed for carbapenem-resistant P. aeruginosa. Random amplified polymorphic DNA (RAPD-PCR) Fingerprinting was used for genotyping the isolates. Results: A total of 80 P. aeruginosa isolates were collected. Isolates were resistant to cefepime 35%, ceftazidime 20%, ciprofloxacin 22%, tazobactam 16%. Out of 80 isolates, 16 were carbapenems-resistant. Gentamicin, tobramycin, and amikacin had the highest susceptibilities of 85%,90%, and 90%, respectively. OprD, blaCTX-M, blaSHV, and blaTEM were detected in 80(100%), 36(45%),22 (27.5%), 17 (21.25%), and 1 (1.25%) blaIMP and blaNDM, respectively. In this study, the blaVIM gene was not detected in the isolates, and no mutation was observed regarding the presence of an insertion element in the OprD gene. Isolates were divided into 13 and 14 common and single types, respectively. Conclusions: P. aeruginosa isolates showed a high rate of β- lactamases production, and the prevalence of carbapenem-resistant, which can be related to different mechanisms, was alarming. On the other hand, the results demonstrated that there was beta-lactam antibiotic resistance and clonal spread among the hospital population. This shows the necessity of molecular surveillance in tracking beta-lactamase-producing strains.

/pdf/evaluation-of-b-lactamases-and-molecular-typing-of-207pc4ju.pdf

Evaluation of β-lactamases and Molecular Typing of Pseudomonas aeruginosa Clinical Strains Isolated from Hospitalized Children in Tehran

This chapter covers five main elements which are: the ions including the divalent cations, the antibiotics particularly those affected by the presence of the divalent cations and the ionophor antibiotics, the enzymes involved in the antibiotic resistance, and the macromolecules and their interaction with the ions. It describes the relationship between the divalent cation and the antibiotic resistance through different related mechanisms. The chapter addresses some case studies concerning the role of the divalent cations in the resistance and the sensitivity of the bacteria during the treatment with the antibiotics. The biological system is sensitive to the chemical structure, particularly the metals. P. aeruginosa is always reported as a multidrug resistant microbe. Divalent cations have an important role in prokaryote cells especially in cell protection or in infection adaptation. A better understanding of the role of the divalent cations in antibiotic resistance will enable better pathogen control. 

The Role of Divalent Cations in Antibiotic Sensitivity

In vitro activity of imipenem/relebactam plus aztreonam against metallo-β-lactamase producing, OprD-deficient Pseudomonas aeruginosa with varying levels of Pseudomonas-derived cephalosporinase production.

The purpose of this study was to analyze the metallo-β-lactamases (MBLs) genotype and oprD mutations of the β-lactam antibiotic-resistant Pseudomonas aeruginosa (PA) strains isolated from southern China. We collected 110 strains of β-lactam antibiotic-resistant PA from 2 hospitals during January 2016–December 2017 from Dongguan, South China. MBLs were detected, amplified, and typed using EDTA disc synergy test, PCR, and Sanger gene sequencing. The mutations and expression levels of oprD were detected using Sanger gene sequencing and qPCR. A total of 16.36% (18/110) β-lactam antibiotic-resistant PA strains produced MBLs, and the main genotypes of MBLs were IMP-25, VIM-2, and SIM-2. Sanger gene sequencing results showed that 107 of the 110 strains harbored mutations in oprD sequence, while 3 strains were negative for oprD amplification (2.73%). Among the 107 strains with positive amplification (97.27%), the rate of intentional mutations (including deletions, insertions, and premature stop codons) was 93.46% (100/107) and that of no disrupted mutation was 6.54% (7/107). qPCR analysis confirmed that the expression level of the OprD protein in the 7 strains of no disrupted mutation was significantly reduced. Among the β-lactam antibiotic-resistant PA strains in southern China, 16.36% were positive for MBLs. The loss rate of oprD was 2.73%, and almost all PA strains showed oprD amplification variation or transcription downregulation. Thus, impaired oprD expression and MBLs production may be some of the mechanisms of β-lactam antibiotic-resistance of PA strains in southern China.

/pdf/analysis-of-metallo-b-lactamases-oprd-mutation-and-multidrug-3a3ribemya.pdf

Analysis of Metallo-β-lactamases, oprD Mutation, and Multidrug Resistance of β-lactam Antibiotic-Resistant Strains of Pseudomonas aeruginosa Isolated from Southern China.

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Analysis of Metallo-β-lactamases, oprD Mutation, and Multidrug Resistance of β-lactam Antibiotic-Resistant Strains of Pseudomonas aeruginosa Isolated from Southern China.