Complete Genome Sequence of Escherichia coli MT102, a Plasmid-Free Recipient Resistant to Rifampin, Azide, and Streptomycin, Used in Conjugation Experiments

Valček, Adam; Overballe‐Petersen, Søren; Hansen, Frank; Dolejská, Monika; Hasman, Henrik

doi:10.1128/mra.00383-19

Cited by 5 publications

(3 citation statements)

References 15 publications

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“…Total cellular DNA from transformants was digested with S1 nuclease and subjected to pulsed-field gel electrophoresis to confirm plasmid carriage. Conjugative transfer of HI2 positive transformants to a plasmid-free, rifampicin and sodium-azide resistant E. coli MT102 recipient strain [ 60 ] was performed using filter-mating method with incubation for 4 h at 28, 30 and 37 °C followed by selection of transconjugants on LB agar plates supplied with cefotaxime (2 mg/L), rifampicin (25 mg/L) and sodium azide (100 mg/L) and incubation overnight at 37 °C. The presence of bla IMP and the HI2 plasmid in four transconjugants was confirmed by PCR [ 34 ] and replicon typing [ 59 ], respectively.…”

Section: Methodsmentioning

confidence: 99%

Epidemic HI2 Plasmids Mobilising the Carbapenemase Gene blaIMP-4 in Australian Clinical Samples Identified in Multiple Sublineages of Escherichia coli ST216 Colonising Silver Gulls

et al. 2021

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Escherichia coli ST216, including those that carry blaKPC-2, blaFOX-5, blaCTX-M-15 and mcr-1, have been linked to wild and urban-adapted birds and the colonisation of hospital environments causing recalcitrant, carbapenem-resistant human infections. Here we sequenced 22 multiple-drug resistant ST216 isolates from Australian silver gull chicks sampled from Five Islands, of which 21 carried nine or more antibiotic resistance genes including blaIMP-4 (n = 21), blaTEM-1b (n = 21), aac(3)-IId (n = 20), mph(A) (n = 20), catB3 (n = 20), sul1 (n = 20), aph(3”)-Ib (n = 18) and aph(6)-Id (n = 18) on FIB(K) (n = 20), HI2-ST1 (n = 11) and HI2-ST3 (n = 10) plasmids. We show that (i) all HI2 plasmids harbour blaIMP-4 in resistance regions containing In809 flanked by IS26 (HI2-ST1) or IS15DI (HI2-ST3) and diverse metal resistance genes; (ii) HI2-ST1 plasmids are highly related to plasmids reported in diverse Enterobacteriaceae sourced from humans, companion animals and wildlife; (iii) HI2 were a feature of the Australian gull isolates and were not observed in international ST216 isolates. Phylogenetic analyses identified close relationships between ST216 from Australian gull and clinical isolates from overseas. E. coli ST216 from Australian gulls harbour HI2 plasmids encoding resistance to clinically important antibiotics and metals. Our studies underscore the importance of adopting a one health approach to AMR and pathogen surveillance.

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Section: Methodsmentioning

confidence: 99%

Epidemic HI2 Plasmids Mobilising the Carbapenemase Gene blaIMP-4 in Australian Clinical Samples Identified in Multiple Sublineages of Escherichia coli ST216 Colonising Silver Gulls

et al. 2021

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“…A conjugation assay based on filter mating method was performed on qnrE1 -positive IncHI2-ST3 (pDR162-CEF-A) and bla CMY-2 -positive IncI1-Iα (pDR162-CEF-B) plasmids. The plasmid-free, rifampicin- and azide-resistant E. coli MT102 ( 42 ) was used as a recipient strain for the conjugative transfer of pDR162-CEF-A and pDR162-CEF-B. Incubation of mating strains was performed on Luri (LB) agar plates at 28°C for pDR162-CEF-A and 37°C for pDR162-CEF-B for 4 h. This was followed by selection of transconjugants on LB plates with ciprofloxacin (0.05 mg/L), rifampicin (25 mg/L), and sodium azide (100 mg/L) for pDR162-CEF-A and cefotaxime (2 mg/L), rifampicin (25 mg/mL), and sodium azide (100 mg/L) for pDR162-CEF-B with incubation overnight at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Clinically relevant antibiotic resistance in Escherichia coli from black kites in southwestern Siberia: a genetic and phenotypic investigation

Tarabai

Krejčí

Karyakin³

et al. 2023

mSphere

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Wild birds including raptors can act as vectors of clinically relevant bacteria with antibiotic resistance. The aim of this study was to investigate the occurrence of antibiotic-resistant Escherichia coli in black kites ( Milvus migrans ) inhabiting localities in proximity to human-influenced environments in southwestern Siberia and investigate their virulence and plasmid contents. A total of 51 E. coli isolates mostly with multidrug resistance (MDR) profiles were obtained from cloacal swabs of 35 (64%, n = 55) kites. Genomic analyses of 36 whole genome sequenced E. coli isolates showed: (i) high prevalence and diversity of their antibiotic resistance genes (ARGs) and common association with ESBL/AmpC production (27/36, 75%), (ii) carriage of mcr-1 for colistin resistance on IncI2 plasmids in kites residing in proximity of two large cities, (iii) frequent association with class one integrase (IntI1, 22/36, 61%), and (iv) presence of sequence types (STs) linked to avian-pathogenic (APEC) and extra-intestinal pathogenic E. coli (ExPEC). Notably, numerous isolates had significant virulence content. One E. coli with APEC-associated ST354 carried qnrE1 encoding fluoroquinolone resistance on IncHI2-ST3 plasmid, the first detection of such a gene in E. coli from wildlife. Our results implicate black kites in southwestern Siberia as reservoirs for antibiotic-resistant E. coli . It also highlights the existing link between proximity of wildlife to human activities and their carriage of MDR bacteria including pathogenic STs with significant and clinically relevant antibiotic resistance determinants. IMPORTANCE Migratory birds have the potential to acquire and disperse clinically relevant antibiotic-resistant bacteria (ARB) and their associated antibiotic resistance genes (ARGs) through vast geographical regions. The opportunistic feeding behavior associated with some raptors including black kites and the growing anthropogenic influence on their natural habitats increase the transmission risk of multidrug resistance (MDR) and pathogenic bacteria from human and agricultural sources into the environment and wildlife. Thus, monitoring studies investigating antibiotic resistance in raptors may provide essential data that facilitate understanding the fate and evolution of ARB and ARGs in the environment and possible health risks for humans and animals associated with the acquisition of these resistance determinants by wildlife.

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“…The fast5 data from ONT MinION platform were base-called, demultiplexed and converted to fastq format using Albacore v2.0.2 (ONT) and adaptor trimmed using Porechop v0.2.2 [ 18 ]. The leftover chromosomal reads were filtered out by mapping the reads on the E. coli TOP10 chromosome using BWA-MEM algorithm [ 19 , 20 ]. The fastq files and trimmed short reads of pHP103 were then used to perform a hybrid assembly using Unicycler v0.4.0 [ 21 ].…”

Section: Methodsmentioning

confidence: 99%

Genomic analysis of qnr-harbouring IncX plasmids and their transferability within different hosts under induced stress

et al. 2022

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Background Conjugative plasmids play a major role in the dissemination of antibiotic resistance genes. Knowledge of the plasmid characteristics and behaviour can allow development of control strategies. Here we focus on the IncX group of plasmids carrying genes conferring quinolone resistance (PMQR), reporting their transfer and persistence within host bacteria of various genotypes under distinct conditions and levels of induced stress in form of temperature change and various concentrations of ciprofloxacin supplementation. Methods Complete nucleotide sequences were determined for eight qnr-carrying IncX-type plasmids, of IncX1 (3), IncX2 (3) and a hybrid IncX1-2 (2) types, recovered from Escherichia coli of various origins. This data was compared with further complete sequences of IncX1 and IncX2 plasmids carrying qnr genes (n = 41) retrieved from GenBank and phylogenetic tree was constructed. Representatives of IncX1 (pHP2) and IncX2 (p194) and their qnrS knockout mutants, were studied for influence of induced stress and genetic background on conjugative transfer and maintenance. Results A high level of IncX core-genome similarity was found in plasmids of animal, environmental and clinical origin. Significant differences were found between the individual IncX plasmids, with IncX1 subgroup plasmids showing higher conjugative transfer rates than IncX2 plasmids. Knockout of qnr modified transfer frequency of both plasmids. Two stresses applied simultaneously were needed to affect transfer rate of wildtype plasmids, whereas a single stress was sufficient to affect the IncX ΔqnrS plasmids. The conjugative transfer was shown to be biased towards the host phylogenetic proximity. A long-term cultivation experiment pointed out the persistence of IncX plasmids in the antibiotic-free environment. Conclusions The study indicated the stimulating effect of ciprofloxacin supplementation on the plasmid transfer that can be nullified by the carriage of a single PMQR gene. The findings present the significant properties and behaviour of IncX plasmids carrying antibiotic resistance genes that are likely to play a role in their dissemination and stability in bacterial populations.

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Complete Genome Sequence of Escherichia coli MT102, a Plasmid-Free Recipient Resistant to Rifampin, Azide, and Streptomycin, Used in Conjugation Experiments

Cited by 5 publications

References 15 publications

Epidemic HI2 Plasmids Mobilising the Carbapenemase Gene blaIMP-4 in Australian Clinical Samples Identified in Multiple Sublineages of Escherichia coli ST216 Colonising Silver Gulls

Epidemic HI2 Plasmids Mobilising the Carbapenemase Gene blaIMP-4 in Australian Clinical Samples Identified in Multiple Sublineages of Escherichia coli ST216 Colonising Silver Gulls

Clinically relevant antibiotic resistance in Escherichia coli from black kites in southwestern Siberia: a genetic and phenotypic investigation

Genomic analysis of qnr-harbouring IncX plasmids and their transferability within different hosts under induced stress

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