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Carbapenemase-producing Enterobacterales (CPEs) pose a considerable threat to public health because of antimicrobial resistance. In France, the most prevalent carbapenemases are OXA-48-like type. More than 55 variants of OXA-48-like enzymes [...]

The Enterobacter cloacae complex (ECC) is highly diverse; its many species and subspecies can be distinguished by using phenotypic methods or matrix-assisted laser desorption/ionization time-offlight mass spectrometry. Whole-genome sequencing enables [...]

Few last-line antimicrobial agents effectively treat infections caused by New Delhi metallo-[beta]-lactamase (NDM)-producing Enterobacterales (1). Cefiderocol is a novel synthetic conjugate siderophore cephalosporin that is more stable against [beta]-lactamase hydrolysis [...]

The massive dissemination of carbapenemase-producing Enterobacterales poses a global threat to public health. Carbapenem antibiotics remain the last line of defense against highly resistant Enterobacterales. Carbapenemases have been identified in [...]

Carbapenems are the last line antimicrobial drugs for treating infections caused by multidrug-resistant Enterobacterales. The global dissemination of carbapenemase-producing Enterobacterales (CPE; formerly known as carbapenemase-producing Enterobacteriaceae) pose a serious threat [...]

In Klebsiella pneumoniae bacteria, resistance to carbapenems results in 2 main mechanisms: the production of an extended spectrum [beta]-lactamase or plasmid-borne cephalosporinase associated with a decrease in permeability of the [...]

This article discusses the emergence of carbapenem resistance in Enterobacterales, specifically through the dissemination of carbapenemase-producing bacteria. The study characterizes two new variants of the VIM-1 enzyme, called VIM-29 and VIM-86, which were isolated in multidrug-resistant Enterobacterales in France. The isolates were resistant to all β-lactam antibiotics except aztreonam/avibactam. The study also analyzes the genetic environment of the VIM-29 and VIM-86 genes and identifies common resistance determinants. The plasmids carrying these genes were found to be self-conjugative. The hydrolytic abilities of VIM-29 and VIM-86 were compared to other prevalent VIM variants, and it was found that they displayed similar resistance profiles. The study concludes that these new variants contribute to the growing problem of carbapenem resistance. [Extracted from the article]

Background Carbapenem-resistant Klebsiella pneumoniae (CRKP) strains are of particular concern, especially strains with mobilizable carbapenemase genes such as bla KPC, bla NDM or bla OXA-48, given that carbapenems are usually the last line drugs in the β-lactam class and, resistance to this sub-class is associated with increased mortality and frequently co-occurs with resistance to other antimicrobial classes. Objectives To characterize the genomic diversity and international dissemination of CRKP strains from tertiary care hospitals in Lisbon, Portugal. Methods Twenty CRKP isolates obtained from different patients were subjected to WGS for species confirmation, typing, drug resistance gene detection and phylogenetic reconstruction. Two additional genomic datasets were included for comparative purposes: 26 isolates (ST13, ST17 and ST231) from our collection and 64 internationally available genomic assemblies (ST13). Results By imposing a 21 SNP cut-off on pairwise comparisons we identified two genomic clusters (GCs): ST13/GC1 (n  = 11), all bearing bla KPC-3, and ST17/GC2 (n  = 4) harbouring bla OXA-181 and bla CTX-M-15 genes. The inclusion of the additional datasets allowed the expansion of GC1/ST13/KPC-3 to 23 isolates, all exclusively from Portugal, France and the Netherlands. The phylogenetic tree reinforced the importance of the GC1/KPC-3-producing clones along with their rapid emergence and expansion across these countries. The data obtained suggest that the ST13 branch emerged over a decade ago and only more recently did it underpin a stronger pulse of transmission in the studied population. Conclusions This study identifies an emerging OXA-181/ST17-producing strain in Portugal and highlights the ongoing international dissemination of a KPC-3/ST13-producing clone from Portugal. [ABSTRACT FROM AUTHOR]

To the Editor: Multidrug resistance in bacteria isolated from animals is an emerging phenomenon, mirroring what is happening among humans. During the past decade, expanded-spectrum [beta]-lactamases in Enterobacteriaceae from humans [...]

While there are numerous variants of CIM that have been proposed [[1]], the "rCIM-A" name has already been published almost a year ago to define a rapid CIM (rCIM) protocol that optimizes results for chromosomal cephalosporinase (AmpC) hyper-producing Enterobacterales [[3]]. The Carbapenem Inactivation Method became "rapid" with the work by Muntean et al. [[6]] with final test results in less than 3 h, a time-to-result similar to those of other rapid CPE confirmation methods (colorimetric, spectrometry, etc.), thus changing the paradigm of CIM testing. As stated by Gauthier et al. interpretation of growth of I E. coli i ATCC 25922 on Mueller-Hinton agar in relation to the antibiotic is not always interpretable at 6 h [[5]]. References 1 Mitteregger D., Wessely J., Barisi'c I., Bedeni'c B., Kosak D., Kundi M. A Variant Carbapenem Inactivation Method (CIM) for Acinetobacter Baumannii Group with Shortened Time-to-Result: RCIM-A. Pathogens. [Extracted from the article]

Background Polymyxins are currently considered a last-resort treatment for infections caused by MDR Gram-negative bacteria. Recently, the emergence of carbapenemase-producing Enterobacteriaceae has accelerated the use of polymyxins in the clinic, resulting in an increase in polymyxin-resistant bacteria. Polymyxin resistance arises through modification of lipid A, such as the addition of phosphoethanolamine (pETN). The underlying mechanisms involve numerous chromosome-encoded genes or, more worryingly, a plasmid-encoded pETN transferase named MCR. Currently, detection of polymyxin resistance is difficult and time consuming. Objectives To develop a rapid diagnostic test that can identify polymyxin resistance and at the same time differentiate between chromosome- and plasmid-encoded resistances. Methods We developed a MALDI-TOF MS-based method, named the MALDIxin test, which allows the detection of polymyxin resistance-related modifications to lipid A (i.e. pETN addition), on intact bacteria, in <15 min. Results Using a characterized collection of polymyxin-susceptible and -resistant Escherichia coli, we demonstrated that our method is able to identify polymyxin-resistant isolates in 15 min whilst simultaneously discriminating between chromosome- and plasmid-encoded resistance. We validated the MALDIxin test on different media, using fresh and aged colonies and show that it successfully detects all MCR-1 producers in a blindly analysed set of carbapenemase-producing E. coli strains. Conclusions The MALDIxin test is an accurate, rapid, cost-effective and scalable method that represents a major advance in the diagnosis of polymyxin resistance by directly assessing lipid A modifications in intact bacteria. [ABSTRACT FROM AUTHOR]

To the Editor: Acinetobacter baumannii is an opportunistic pathogen that is a source of nosocomial infections, mostly pneumonia (7). Treatment of infections caused by A. baumannii is becoming a serious [...]

Background: There is an urgent need for accurate and rapid diagnostic tests to identify carbapenemase producing enterobacteria (CPE). Here, we have evaluated the Carbapenem Inactivation Method (CIM) test to detect CPEs from cultured colonies. Methods: A total of 256 enterobacterial isolates were used to evaluate the performance of the CIM in comparison to Carba NP test and molecular detection used a reference method. Ninety three well-characterized isolates (including 29 non-CPE and 63 CPEs of worldwide origin) with decreased susceptibility to at least one carbapenem were used to (i) evaluate the efficacy of CIM test and (ii) to compare it to the Carba NP test. We also tested different carbapenems to determine the best substrate for this test. Finally, the CIM test was then evaluated prospectively against 164 isolates referred to the French National Reference Center (NRC) for Antimicrobial Resistance from may 2016 to july 2016. Results: Based on the results of this retrospective study, sensitivity and specificity of the CIM and the Carba NP test were 92.1% and 100%, respectively. We demonstrated that the meropenem was the best substrate to perform the CIM test since sensitivity and specificity were 81.1% and 100% using ertapenem disk, and 100% and 65,6% using imipenem disk, and respectively. Taking in account the results of retrospective and prospective studies, CIM and Carba NP tests have similar sensitivity, specificity, positive predictive value and negative predictive values being 96.3%, 98.9%, 99.0% and 98.4% for the CIM test versus 96.9%, 100%, 100% and 100% for the Carba NP test. Conclusions: Our results confirm that the CIM test may be a useful tool for the reliable confirmation of carbapenemase-activity in enterobacterial isolates, especially in clinical microbiological laboratories with limited resources, no trained personnel, and no specialized equipment. [ABSTRACT FROM AUTHOR]

Cefiderocol is a novel siderophore cephalosporin, which has proven in vitro activity against carbapenem-resistant (CR) Gram-negative pathogens and stability towards all carbapenemases. The aim of this study was to describe the first cases of prescriptions and the efficacy of cefiderocol for compassionate use in the 2 months following its access in France. We performed a national retrospective study of all patients who received at least one dose of cefiderocol from 2 November 2018 to 5 November 2019. We collected clinical characteristics and outcome through a standard questionnaire. Bacterial isolates from 12 patients were centralized and analyzed in the French National Reference Center for Antimicrobial Resistance, and sequenced using Illumina technology. Finally, 13 patients from 7 French university hospitals were included in the study. The main type of infection treated by cefiderocol was respiratory tract infections (RTI, n = 10). The targeted bacteria were Pseudomonas aeruginosa (n = 12), including carbapenemase-producing P. aeruginosa (n = 9), Acinetobacter baumannii (n = 2), Klebsiella pneumoniae (n = 1), and Enterobacter hormaechei (n = 1). Overall, of the 12 patients whose samples were analyzed, 5 P. aeruginosa strains were not susceptible to cefiderocol (4 categorized as resistant and 1 as intermediate) according to Clinical and Laboratory Standards Institute (CLSI) breakpoints. If considering susceptible strains, the cure rate was 6/7, while being 0/5 among not-susceptible strains. This study underlines the necessity to test strains in adequate conditions. [ABSTRACT FROM AUTHOR]

[english] Carbapenem-resistant are increasingly reported worldwide and cause therapeutic problem in health care facilities. In this study 28 imipenem-resistant were examined for expression of carbapenemases by phenotypic and genotypic methods. Modified Hodge Test (MHT), CarbaNP test were used for phenotypic detection, and PCR using specific primers for the detection of and type carbapenemases with specific primers were performed. MHT and CarbaNP tests were positive for all of imipenem-resistant . The gene was detected in 27/28 isolates. One isolate was positive for the presence of the gene. According to our results NP test and MHT have high sensitivity and specificity for detection of those carbapenemases. This study reports the first cases of OXA-48-producing in Iran.

Highlights • Development of a rapid and reliable multiplex polymerase chain reaction for mcr -1 to mcr- 5 gene detection. • Sensitivity and specificity of 100% using a collection of mcr -positive isolates. • Prospective evaluation revealed 100% positive and negative predictive values. ABSTRACT Plasmid-mediated colistin resistance is increasingly described worldwide in Enterobacteriaceae from animal and human isolates. Diffusion of these resistance traits among carbapenem-resistant enterobacterial isolates is of particular concern as colistin has become the last resort antibiotic for treating human infections with these organisms. Therefore, being able to monitor the presence of these transferable colistin resistance genes (mcr-1 to mcr-5 -variants) is crucial. This paper describes the development of a multiplex polymerase chain reaction (PCR) protocol for detection of all currently known transferable colistin resistance genes in Enterobacteriaceae. Five primer pairs were designed to amplify mcr-1, mcr-2, mcr-3, mcr-4 and mcr -5 gene products in a multiplex PCR. This assay was validated retrospectively on colonies of 50 Escherichia coli , 44 Klebsiella pneumoniae and 12 Salmonella enterica isolates of animal and human origin, all well characterized, and validated prospectively on 450 carbapenem-resistant enterobacterial isolates received by the French National Reference Centre. In addition, 82 Aeromonas spp. and 10 Shewanella spp. known to be the progenitors of mcr-3 and mcr-4 alleles, respectively, were screened. Mcr-multiplex PCR assay displayed 100% specificity, sensitivity, negative predictive value and positive predictive value. The assay was able to detect all variants of the different mcr alleles, and was able to detect chromosomally encoded mcr-4- like variants present in two Shewanella bicestrii JAB-1 and Shewanella woodyi S539. In conclusion, a rapid and robust multiplex PCR assay able to detect all known mcr gene families described in Enterobacteriaceae was developed and validated. This type of test is critical for the epidemiological surveillance of plasmid-encoded resistance, especially in carbapenem-resistant bacteria. [ABSTRACT FROM AUTHOR]

OXA-48-producing Enterobacterales have now widely disseminated throughout the world. Several variants have now been reported, differing by just a few amino-acid substitutions or deletions, mostly in the region of the loop β5-β6. As OXA-48 hydrolyzes carbapenems but lacks significant expanded-spectrum cephalosporin (ESC) hydrolytic activity, ESCs were suggested as a therapeutic option. Here, we have characterized OXA-517, a natural variant of OXA-48- with an Arg214Lys substitution and a deletion of Ile215 and Glu216 in the β5-β6 loop, capable of hydrolyzing at the same time ESC and carbapenems. MICs values of E. coli expressing bla OXA-517 gene was located on a ca. 31-kb plasmid identical to the prototypical IncL bla OXA-517 gene was located on a ca. 31-kb plasmid identical to the prototypical IncL bla OXA-48 -carrying plasmid except for an IS 1R -mediated deletion of 30.7-kb in the tra operon. The crystal structure of OXA-517, determined to 1.86 Å resolution, revealed an expanded active site compared to that of OXA-48, which allows for accommodation of the bulky ceftazidime substrate. Our work illustrates the remarkable propensity of OXA-48-like carbapenemases to evolve through mutation/deletion in the β5-β6 loop to extend its hydrolysis profile to encompass most β-lactam substrates.

A multidrug-resistant Klebsiella pneumoniae 1210 isolate with reduced carbapenem susceptibility revealed the presence of a novel plasmid-encoded bla OXA-48-like The 60.7-kb plasmid (pOXA-519) was similar to the IncL-OXA-48 prototypical plasmid except for a ca. 2-kb deletion due to an IS bla insertion. OXA-519 differed from OXA-48 by a Val120Leu substitution, which resulted in an overall reduced β-lactam-hydrolysis profile, except those for ertapenem and meropenem, which were increased. Thus, detection of OXA-519 producers using biochemical tests that monitor imipenem hydrolysis will be difficult.OXA-519 The 60.7-kb plasmid (pOXA-519) was similar to the IncL-OXA-48 prototypical plasmid except for a ca. 2-kb deletion due to an IS 1R insertion. OXA-519 differed from OXA-48 by a Val120Leu substitution, which resulted in an overall reduced β-lactam-hydrolysis profile, except those for ertapenem and meropenem, which were increased. Thus, detection of OXA-519 producers using biochemical tests that monitor imipenem hydrolysis will be difficult., (Copyright © 2018 American Society for Microbiology.)