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Objective: Antimicrobial resistance (AMR), together with multidrug resistance (MDR), mainly among Gram-negative bacteria, has been on the rise. Colistin (polymyxin E) remains one of the primary available last resorts to treat infections caused by MDR bacteria during the rapid emergence of global resistance. As the exact mechanism of bacterial resistance to colistin remains undetermined, this study warranted elucidation of the underlying mechanisms of colistin resistance and heteroresistance among carbapenem-resistant Klebsiella pneumoniae isolates., Methods: Molecular analysis was carried out on the resistant isolates using a genome-wide characterisation approach, as well as MALDI-TOF mass spectrometry, to identify lipid A., Results: Among the 32 carbapenem-resistant K. pneumoniae isolates, several isolates showed resistance and intermediate resistance to colistin. The seven isolates with intermediate resistance exhibited the "skip-well" phenomenon, attributed to the presence of resistant subpopulations. The three isolates with full resistance to colistin showed ions using MALDI-TOF mass spectrometry at m/z of 1840 and 1824 representing bisphosphorylated and hexa-acylated lipid A, respectively, with or without hydroxylation at position C'-2 of the fatty acyl chain. Studying the genetic environment of mgrB locus revealed the presence of two insertion sequences that disrupted the mgrB locus in the three colistin-resistant isolates: IS1R and IS903B., Conclusions: Our findings show that colistin resistance/heteroresistance was inducible with mutations in chromosomal regulatory networks controlling the lipid A moiety and insertion sequences disrupting the mgrB gene, leading to elevated minimum inhibitory concentration values and treatment failure. Different treatment strategies should be employed to avoid colistin heteroresistance-linked treatment failures, mainly through combination therapy using colistin with carbapenems, aminoglycosides, or tigecycline., Competing Interests: Declaration of competing interests The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

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., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Enterobacter cloacae complex (ECC) members are rapidly emerging as successful nosocomial pathogens, especially, with the emergence of carbapenem-resistant clones. In this study, we performed a comprehensive molecular characterization of a carbapenem-resistant E. hormaechei ssp. xiangfangensis LAU_ENC1. hsp60 and average nucleotide identity (ANI) were used for its identification. The repertoire of resistance genes and phage content were analyzed. Plasmid sequences were extracted and compared to closest references. The isolate LAU_ENC1 was identified as an E. hormaechei ssp. xiangfangensis and belonged to ST-114A sub-cluster. bla NDM-1 , bla CTX-M-15 , bla OXA-1 , and bla ACT-16 genes were detected as β-lactam resistance determinants. A chromosomal hybrid intact phage, Enterobacter phage LAU1, with bla CTX-M-15 integrated in its direct vicinity within an ISEcp1 - bla CTX-M-15 - wbuC - ∆Tn2 rare cassette was detected. bla NDM-1 was integrated within a novel IncFII conjugative plasmid, pLAU_ENC1, through an IS3000- ΔISAba125-bla NDM-1 -ble MBL -//-Tn5403 cassette. To our knowledge, this is the first report of a multi-drug resistant (MDR) E. hormaechei ssp. xiangfangensis carrying a bla CTX-M-15 integrated within the proximity of a provirus chromosomal region. Treatment options for MDR ECC members are becoming scarce, thus warranting an increased monitoring of the dissemination of these pathogens in clinical settings., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The prevalence of antibiotic-resistant bacteria in surface water in Lebanon is a growing concern and understanding the mechanisms of the spread of resistance determinants is essential. We aimed at studying the occurrence of resistant bacteria and determinants in surface water sources in Lebanon and understanding their mobilization and transmission. Water samples were collected from five major rivers in Lebanon. A total of 91 isolates were recovered by incubating at 37°C on Blood and MacConkey agar out of which 25 were multi-drug resistant (MDR) and accordingly were further characterized. Escherichia coli and Klebsiella pneumoniae were the most common identified MDR isolates. Conjugation assays coupled with in silico plasmid analysis were performed and validated using PCR-based replicon typing (PBRT) to identify and confirm incompatibility groups and the localization of β-lactamase encoding genes. Escherichia coli EC23 carried a blaNDM-5 gene on a conjugative, multireplicon plasmid, while blaCTX-M-15 and blaTEM-1B were detected in the majority of the MDR isolates. Different sequence types (STs)were identified including the highly virulent E. coli ST131. Our results showed a common occurrence of bacterial contaminants in surface water and an increase in the risk for the dissemination of resistance determinants exacerbated with the ongoing intensified population mobility in Lebanon and the widespread lack of wastewater treatment., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Carbapenem-resistant Acinetobacter baumannii (CRAB) is an important opportunistic pathogen linked to a variety of nosocomial infections and hospital outbreaks worldwide. This study aimed at investigating and characterizing a CRAB outbreak at a large tertiary hospital in Lebanon. A total of 41 isolates were collected and analyzed using pulsed-field gel electrophoresis (PFGE). Whole-genome sequencing (WGS) was performed on all the isolates, and long-read PacBio sequencing was used to generate reference genomes. The multilocus sequence types (MLST), repertoire of resistance genes, and virulence factors were determined from the sequencing data. The plasmid content was analyzed both in silico and using the A. baumannii PCR-based replicon typing (AB-PBRT) method. Genome analysis initially revealed two clones, one carrying bla OXA-23 on Tn 2006 (ST-1305, ST-195, and ST-218) and another carrying bla OXA-72 on pMAL-1 (ST-502 and ST-2059, a new ST), with the latter having two subclones, as revealed using the Bayesian transmission network. All isolates were extensively drug resistant (XDR). WGS analysis revealed the transmission pathways and demonstrated the diversity of CRAB isolates and mobile genetic elements in this health care setting. Outbreak detection using WGS and immediate implementation of infection control measures contribute to restraining the spread and decreasing mortality. IMPORTANCE Carbapenem-resistant Acinetobacter baumannii (CRAB) has been implicated in hospital outbreaks worldwide. Here, we present a whole-genome-based investigation of an extensively drug-resistant CRAB outbreak rapidly spreading and causing high incidences of mortality at numerous wards of a large tertiary hospital in Lebanon. This is the first study of its kind in the region. Two circulating clones were identified using a combination of molecular typing approaches, short- and long-read sequencing and Bayesian transmission network analysis. One clone carried bla OXA-23 on Tn 2006 (ST-1305, ST-195, and ST-218), and another carried bla OXA-72 on a pMAL-1 plasmid (ST-502 and ST-2059, a new ST). A pMAL-2 plasmid was circulating between the two clones. The approaches implemented in this study and the obtained findings facilitate the tracking of outbreak scenarios in Lebanon and the region at large., (Copyright © 2020 Makke et al.)

Introduction: Over the past decade methicillin-resistant Staphylococcus aureus (MRSA) has been recognized as a major cause of healthcare associated infections. Recently, however, epidemiology of this pathogen has changed drastically with the emergence of new clones in the community. Efficient epidemiological typing methods are essential to monitor and limit the occurrence and spread of epidemic clones., Methodology: A total of sixty S. aureus isolates were collected from the Jordan University hospital in Amman-Jordan. All isolates were characterized using Staphylococcus protein A (spa) typing and pulsed-field gel electrophoresis (PFGE). Samples were tested for their susceptibility patterns against seven antimicrobial agents and for their potential to form biofilms., Results: spa typing showed that spa type t044 was the most common representing 28% of the isolates studied and 38% of the MRSA population. PFGE revealed fourty-six pulsotypes among the sixty tested isolates clustering similar spa types together. The predominant resistance was detected against levofloxacin, chloramphenicol and clindamycin. One MSSA isolate typed as spa t955 showed biofilm formation potential through protein deposition.., Conclusion: The study results are based on one hospital, but the findings of this and other studies conducted in the region indicate that there is an urgent need for standardized surveillances combined with the application of well-validated typing methods to assess the occurrence of MRSA and to control its spread.

Objectives: The occurrence and dissemination of methicillin-resistant Staphylococcus aureus (MRSA) in healthcare settings and the community and its risk of being introduced into hospitals are matters of great concern. The purpose of this study was to conduct a miniaturized epidemiological analysis of S. aureus-associated infections and to characterize the isolates by a variety of molecular typing techniques. Ongoing molecular surveillance is essential to prevent S. aureus strains from becoming endemic in the Lebanese healthcare setting., Methods: A total of 132 S. aureus from different clinical specimens were isolated over a 6-month period. Characterization of the isolates was done by detection of the mecA gene, Panton-Valentine leukocidin determinant detection, staphylococcal chromosomal cassette (SCCmec) typing of MRSA, S. aureus protein A (spa) typing, multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and antibiogram analysis., Results: MRSA represented 30% of the isolates, with PVL being detected in 54% of MRSA and 12% of methicillin-susceptible S. aureus (MSSA). A difference between MRSA and MSSA was observed in the spa types. Clustering SCCmec with MLST identified seven MRSA and 20 MSSA clones, with PVL-positive ST80-MRSA-IV being the dominant clone (7%), while PFGE revealed 32 groups with 80% cutoff similarity., Conclusions: Although the results of this study are based on samples collected from one hospital, the high diversity observed along with the lack of any equivalence in the genetic backgrounds of the major MSSA and MRSA clones, emphasizes the urgent need for standardized surveillance combined with the application of well-validated typing methods to assess the occurrence of MRSA and subsequently to control its spread., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

AimExtended-spectrum β-lactamase-producing (ESBL) Escherichia coli are a public threat worldwide. This study aimed at analyzing the genomic and functional attributes of nine ESBLs taken from rectal swabs.Materials & methodsSamples were isolated from patients admitted for gastrointestinal and urological procedures at the University Medical Center-Rizk Hospital (UMCRH) in Lebanon. Illumina paired-end libraries were prepared and sequenced.ResultsThe isolates were distributed into five lineages: ST131, ST648, ST405, ST73 and ST38, and harbored bla OXA-1, bla TEM-1B, bla TEM-1C and aac(6')Ib-cr. ST131 isolates were carriers of stx2 converting I phage.ConclusionThis is the first comprehensive genomic analysis performed on ESBLs in Lebanon.

AimExtended-spectrum β-lactamase-producing (ESBL) Escherichia coli are a public threat worldwide. This study aimed at analyzing the genomic and functional attributes of nine ESBLs taken from rectal swabs.Materials & methodsSamples were isolated from patients admitted for gastrointestinal and urological procedures at the University Medical Center-Rizk Hospital (UMCRH) in Lebanon. Illumina paired-end libraries were prepared and sequenced.ResultsThe isolates were distributed into five lineages: ST131, ST648, ST405, ST73 and ST38, and harbored bla OXA-1, bla TEM-1B, bla TEM-1C and aac(6')Ib-cr. ST131 isolates were carriers of stx2 converting I phage.ConclusionThis is the first comprehensive genomic analysis performed on ESBLs in Lebanon.