Linkage of determinants for streptogramin A, macrolide-lincosamide-streptogramin B, and chloramphenicol resistance on a conjugative plasmid in Enterococcus faecium and dissemination of this cluster among streptogramin-resistant enterococci

2000 ◽  
Vol 290 (6) ◽  
pp. 543-548 ◽  
Author(s):  
Guido Werner ◽  
Bianca Hildebrandt ◽  
Ingo Klare ◽  
Wolfgang Witte
Keyword(s):  
Enterococcus Faecium ◽  
Conjugative Plasmid ◽  
Chloramphenicol Resistance

scholarly journals The Enterococcus Cassette Chromosome, a Genomic Variation Enabler in Enterococci

mSphere ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
A. Sivertsen ◽  
J. Janice ◽  
T. Pedersen ◽  
T. M. Wagner ◽  
J. Hegstad ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Enterococcus Faecium ◽  
Genomic Variation ◽  
Conjugative Plasmid ◽  
Natural Habitats ◽  
Chloramphenicol Resistance ◽  
Content Type ◽  
Serine Recombinases ◽  
Accessory Genes

ABSTRACT Enterococcus faecium has a highly variable genome prone to recombination and horizontal gene transfer. Here, we have identified a novel genetic island with an insertion locus and mobilization genes similar to those of staphylococcus cassette chromosome elements SCCmec. This novel element termed the enterococcus cassette chromosome (ECC) element was located in the 3′ region of rlmH and encoded large serine recombinases ccrAB similar to SCCmec. Horizontal transfer of an ECC element termed ECC::cat containing a knock-in cat chloramphenicol resistance determinant occurred in the presence of a conjugative reppLG1 plasmid. We determined the ECC::cat insertion site in the 3′ region of rlmH in the E. faecium recipient by long-read sequencing. ECC::cat also mobilized by homologous recombination through sequence identity between flanking insertion sequence (IS) elements in ECC::cat and the conjugative plasmid. The ccrABEnt genes were found in 69 of 516 E. faecium genomes in GenBank. Full-length ECC elements were retrieved from 32 of these genomes. ECCs were flanked by attR and attL sites of approximately 50 bp. The attECC sequences were found by PCR and sequencing of circularized ECCs in three strains. The genes in ECCs contained an amalgam of common and rare E. faecium genes. Taken together, our data imply that ECC elements act as hot spots for genetic exchange and contribute to the large variation of accessory genes found in E. faecium. IMPORTANCE Enterococcus faecium is a bacterium found in a great variety of environments, ranging from the clinic as a nosocomial pathogen to natural habitats such as mammalian intestines, water, and soil. They are known to exchange genetic material through horizontal gene transfer and recombination, leading to great variability of accessory genes and aiding environmental adaptation. Identifying mobile genetic elements causing sequence variation is important to understand how genetic content variation occurs. Here, a novel genetic island, the enterococcus cassette chromosome, is shown to contain a wealth of genes, which may aid E. faecium in adapting to new environments. The transmission mechanism involves the only two conserved genes within ECC, ccrABEnt, large serine recombinases that insert ECC into the host genome similarly to SCC elements found in staphylococci.

Studies on the role of IS1216E in the formation and dissemination of poxtA-carrying plasmids in an Enterococcus faecium clade A1 isolate

2020 ◽  
Vol 75 (11) ◽  
pp. 3126-3130
Author(s):  
Xinxin Shan ◽  
Xin-Sheng Li ◽  
Nannan Wang ◽  
Stefan Schwarz ◽  
Su-Mei Zhang ◽  
...  
Keyword(s):  
Enterococcus Faecium ◽  
Blot Hybridization ◽  
Tetracycline Resistance ◽  
Southern Blot Hybridization ◽  
Conjugative Plasmid ◽  
Fusion Event ◽  
Mobilizable Plasmid ◽  
Antimicrobial Resistance Genes ◽  
Positive Isolate

Abstract Objectives To analyse the role of IS1216E in the dissemination of the phenicol-oxazolidinone-tetracycline resistance gene poxtA in an Enterococcus faecium clade A1 isolate. Methods MICs were determined by broth microdilution. The poxtA-positive isolate was typed by MLST. The two plasmids were characterized by PCR, conjugation, S1-PFGE, Southern blot hybridization and WGS analysis. The presence of translocatable units (TUs) was examined by PCR and sequencing. Results Isolate E1077 contains the 217661 bp conjugative plasmid pE1077-217 and the 23710 bp mobilizable plasmid pE1077-23. pE1077-217 harbours erm(B), aac(A)-aph(D), aadE, spw, lsa(E), lnu(B), aphA3 and dfrG, whereas pE1077-23 carries a Tn6657-like transposon containing poxtA and fexB. pE1077-23 was apparently formed by an IS1216E-mediated composite transposon–plasmid fusion event, involving a replicative transposition process. Conjugation experiments showed that pE1077-23 is mobilizable by pE1077-217. Moreover, a novel 31742 bp plasmid, pT-E1077-31, was found in a transconjugant. WGS analysis indicated that pT-E1077-31 was formed by the integration of a Tn6657-derived, IS1216E-based translocatable unit, which carried fexB and poxtA, into a copy of pE1077-23. Conclusions This study showed the presence of two cointegrate formation events in the formation and spread of a poxtA/fexB-carrying plasmid in E. faecium. One was the integration of a transposon into a plasmid while the other was the integration of a TU into a different site of the same type of plasmid-borne transposon from which it originated. In both events, IS1216E played a major role, suggesting that IS1216E-mediated transposition and translocation processes aid the dissemination and persistence of important antimicrobial resistance genes, such as poxtA, among enterococci.

scholarly journals tcrB, a Gene Conferring Transferable Copper Resistance in Enterococcus faecium: Occurrence, Transferability, and Linkage to Macrolide and Glycopeptide Resistance

2002 ◽  
Vol 46 (5) ◽  
pp. 1410-1416 ◽  
Author(s):  
Henrik Hasman ◽  
Frank M. Aarestrup
Keyword(s):  
Enterococcus Faecium ◽  
Antimicrobial Agents ◽  
Brain Heart Infusion ◽  
Copper Resistance ◽  
Conjugative Plasmid ◽  
Feed Additive ◽  
Strongly Correlated ◽  
Glycopeptide Resistance ◽  
Copper Resistance Gene ◽  
Infusion Agar

ABSTRACT A newly discovered gene, designated tcrB, which is located on a conjugative plasmid conferring acquired copper resistance in Enterococcus faecium, was identified in an isolate from a pig. The tcrB gene encodes a putative protein belonging to the CPx-type ATPase family with homology (46%) to the CopB protein from Enterococcus hirae. The tcrB gene was found in E. faecium isolated from pigs (75%), broilers (34%), calves (16%), and humans (10%) but not in isolates from sheep. Resistant isolates, containing the tcrB gene, grew on brain heart infusion agar plates containing up to 28 mM CuSO4 compared to only 4 mM for the susceptible isolates. Copper resistance, and therefore the presence of the tcrB gene, was strongly correlated to macrolide and glycopeptide resistance in isolates from pigs, and the tcrB gene was shown to be located on the same conjugative plasmid as the genes responsible for resistance to these two antimicrobial agents. The frequent occurrence of this new copper resistance gene in isolates from pigs, where copper sulfate is being used in large amounts as feed additive, suggests that the use of copper has selected for resistance.

scholarly journals Conjugative transfer of high-level mupirocin resistance from Staphylococcus haemolyticus to other staphylococci.

1997 ◽  
Vol 41 (3) ◽  
pp. 693-695 ◽  
Author(s):  
E E Udo ◽  
L E Jacob ◽  
E M Mokadas
Keyword(s):  
Staphylococcus Aureus ◽  
Laboratory Strain ◽  
Conjugative Plasmid ◽  
Aureus Strain ◽  
Conjugative Transfer ◽  
Coagulase Negative Staphylococci ◽  
Chloramphenicol Resistance ◽  
Staphylococcus Haemolyticus ◽  
Mupirocin Resistance ◽  
High Level

A conjugative plasmid, pXU10, encoding high-level mupirocin resistance was transferred from a Staphylococcus haemolyticus isolate, CN216, to other coagulase-negative staphylococci and a restriction deficient Staphylococcus aureus strain, XU21, but not to clinical isolates or a restriction-proficient laboratory strain (strain WBG541) of S. aureus. However, from XU21 it was cotransferred with a 3.5-kb chloramphenicol resistance plasmid to WBG541. The results demonstrated the ability of pXU10 to mobilize nonconjugative plasmids.

Linkage of a novel mercury resistance operon with streptomycin resistance on a conjugative plasmid in Enterococcus faecium

Plasmid ◽  
2005 ◽  
Vol 54 (1) ◽  
pp. 26-38 ◽  
Author(s):  
Ian J. Davis ◽  
Adam P. Roberts ◽  
Derren Ready ◽  
Hilary Richards ◽  
Michael Wilson ◽  
...  
Keyword(s):  
Enterococcus Faecium ◽  
Streptomycin Resistance ◽  
Conjugative Plasmid ◽  
Mercury Resistance

scholarly journals Presence of a vanA-Carrying Pheromone Response Plasmid (pBRG1) in a Clinical Isolate of Enterococcus faecium

2003 ◽  
Vol 47 (5) ◽  
pp. 1571-1576 ◽  
Author(s):  
Gloria Magi ◽  
Roberta Capretti ◽  
Claudia Paoletti ◽  
Marco Pietrella ◽  
Luigi Ferrante ◽  
...  
Keyword(s):  
Sex Pheromone ◽  
Clinical Isolate ◽  
Enterococcus Faecium ◽  
Conjugative Plasmid ◽  
Pheromone Response ◽  
Cell Infection ◽  
Cell Monolayers ◽  
Aggregation Substance ◽  
Pcr Products ◽  
The One

ABSTRACT Sex pheromone plasmids, frequently found in Enterococcus faecalis, have rarely been detected in Enterococcus faecium. pBRG1 is an approximately 50-kb vanA-carrying conjugative plasmid of an E. faecium clinical isolate (LS10) that is transferable to E. faecalis laboratory strains. In cell infection experiments, E. faecium LS10 exhibited remarkably high invasion efficiency and produced cytopathogenic effects in Caco-2 cell monolayers. Growth in the presence of sex pheromones produced by E. faecalis JH2-2 was found to cause self-aggregation of both E. faecium LS10 and E. faecalis JH-RFV(pBRG1) (a transconjugant obtained by transfer of pBRG1 to E. faecalis JH2-2) and to increase the cell adhesion and invasion efficiencies of both E. faecium LS10 and E. faecalis JH-RFV(pBRG1). Sex pheromone cCF10 caused clumping of E. faecalis OG1RF(pBRG1) (a transconjugant obtained by transfer of pBRG1 to E. faecalis OG1RF) at a concentration ∼100-fold higher than the one required for the control strain E. faecalis OG1RF(pCF10). PCR products of the expected sizes were obtained with primers internal to aggregation substance genes of E. faecalis pheromone response plasmids pAD1, pPD1, and pCF10 and primers internal to ash701 of E. faecium pheromone plasmid pHKK701. These findings suggest that pBRG1 of E. faecium LS10 is a sex pheromone response plasmid.

IS26 cannot move alone

2021 ◽  
Author(s):  
Christopher J Harmer ◽  
Ruth M Hall
Keyword(s):  
Insertion Sequence ◽  
Conjugative Plasmid ◽  
Gram Negative Bacteria ◽  
Gibson Assembly ◽  
Transposase Gene ◽  
Chloramphenicol Resistance ◽  
Resistance Determinants ◽  
End Products ◽  
Insertion Sequence Is26 ◽  
Plasmid Backbone

Abstract Background IS26 plays a major role in the dissemination of antibiotic resistance determinants in Gram-negative bacteria. Objectives To determine whether insertion sequence IS26 is able to move alone (simple transposition) or if it exclusively forms cointegrates. Methods A two-step PCR using outward-facing primers was used to search for circular IS26 molecules. Gibson assembly was used to clone a synthetic IS26 containing a catA1 chloramphenicol resistance gene downstream of the tnp26 transposase gene into pUC19. IS activity in a recA−  Escherichia coli containing the non-conjugative pUC19-derived IS26::catA1 construct and the conjugative plasmid R388 was detected using a standard mating-out assay. Transconjugants were screened for resistance. Results Circular IS26 molecules that would form with a copy-out route were not detected by PCR. The synthetic IS26::catA1 construct formed CmRTpR transconjugants (where CmR and TpR stand for chloramphenicol resistant and trimethoprim resistant, respectively), representing an R388 derivative carrying the catA1 gene at a frequency of 5.6 × 10−7 CmRTpR transconjugants per TpR transconjugant, which is comparable to the copy-in activity of the unaltered IS26. To test for simple transposition of IS26::catA1 (without the plasmid backbone), 1200 CmRTpR colonies were screened and all were resistant to ampicillin, indicating that the pUC19 backbone was present. Hence, IS26::catA1 had only formed cointegrates. Conclusions IS26 is unable to move alone and cointegrates are the exclusive end products of the reactions mediated by the IS26 transposase Tnp26. Consequently, when describing the formation of complex resistance regions, simple ‘transposition’ of a single IS26 should not be invoked.

Metallo-beta-lactamase-producing Escherichia coli in the sewage of Mexico City: where do they come from?

2021 ◽  
Author(s):  
Kathia Lüneberg ◽  
Carlos F. Amabile-Cuevas ◽  
Eduardo Mucito-Varela ◽  
Leticia Martínez ◽  
Eva Salinas ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mexico City ◽  
Municipal Wastewater ◽  
Conjugative Plasmid ◽  
Beta Lactamase ◽  
Chloramphenicol Resistance ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Genes Encoding ◽  
Wastewater Bacteria

While monitoring the presence of antibiotic resistance in municipal wastewater bacteria from Mexico City, five Escherichia coli isolates were detected to be resistant to carbapenems, antibiotics of “last resort” used mostly in hospitals. Further analysis revealed that these carbapenem-resistant isolates carried the gene for a metallo-beta-lactamase, NDM-5. The gene was found to be beared by a large, ~145 kb conjugative plasmid, which also carries putative genes encoding resistance to sulfonamides, trimethoprim, tetracycline, ciprofloxacin, chloramphenicol (although no phenotypic chloramphenicol resistance was detected) and quaternary-ammonium compounds. The plasmid also carried gene mobility determinants, such as an integron integrase, and two transposases. In addition to the direct public health threat posed by the presence of such multi-resistant organisms in wastewater released into the environment and used for crop irrigation; it is particularly concerning that carbapenem-resistant E. coli is rather rare in Mexican hospitals (<1%), but was found in small, 100-mL samples of municipal wastewater. This could suggest that, either these organisms are under-reported by clinical microbiology laboratories, underlining the usefulness of wastewater monitoring; or that there is an unknown source of such carbapenem-resistant organisms that are being dumped into the wastewater. The source of these bacteria must be assessed and controlled to prevent the further spread of this multi-resistance plasmid among other environmental and clinical microorganisms.

scholarly journals Linezolid resistance in Enterococcus faecium and Enterococcus faecalis from hospitalized patients in Ireland: high prevalence of the MDR genes optrA and poxtA in isolates with diverse genetic backgrounds

2020 ◽  
Vol 75 (7) ◽  
pp. 1704-1711 ◽  
Author(s):  
Sarah A Egan ◽  
Anna C Shore ◽  
Brian O’Connell ◽  
Grainne I Brennan ◽  
David C Coleman
Keyword(s):  
Enterococcus Faecalis ◽  
Resistance Genes ◽  
Enterococcus Faecium ◽  
Illumina Miseq ◽  
Conjugative Plasmid ◽  
Sequence Coverage ◽  
Linezolid Resistance ◽  
Oxford Nanopore ◽  
High Prevalence ◽  
Mdr Genes

Abstract Objectives To investigate the prevalence of the optrA, poxtA and cfr linezolid resistance genes in linezolid-resistant enterococci from Irish hospitals and to characterize associated plasmids. Methods One hundred and fifty-four linezolid-resistant isolates recovered in 14 hospitals between June 2016 and August 2019 were screened for resistance genes by PCR. All isolates harbouring resistance genes, and 20 without, underwent Illumina MiSeq WGS. Isolate relatedness was assessed using enterococcal whole-genome MLST. MinION sequencing (Oxford Nanopore) and hybrid assembly were used to resolve genetic environments/plasmids surrounding resistance genes. Results optrA and/or poxtA were identified in 35/154 (22.7%) isolates, the highest prevalence reported to date. Fifteen isolates with diverse STs harboured optrA only; one Enterococcus faecium isolate harboured optrA (chromosome) and poxtA (plasmid). Seven Enterococcus faecalis and one E. faecium harboured optrA on a 36 331 bp plasmid with 100% identity to the previously described optrA-encoding conjugative plasmid pE349. Variations around optrA were also observed, with optrA located on plasmids in five isolates and within the chromosome in three isolates. Nine E. faecium and 10 E. faecalis harboured poxtA, flanked by IS1216E, within an identical 4001 bp region on plasmids exhibiting 72.9%–100% sequence coverage to a 21 849 bp conjugative plasmid. E. faecalis isolates belonged to ST480, whereas E. faecium isolates belonged to diverse STs. Of the remaining 119 linezolid-resistant isolates without linezolid resistance genes, 20 investigated representatives all harboured the G2576T 23S RNA gene mutation associated with linezolid resistance. Conclusions This high prevalence of optrA and poxtA in diverse enterococcal lineages in Irish hospitals indicates significant selective pressure(s) for maintenance.

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