ABSTRACT
OBJECTIVES: To characterize blaIMP-4-carrying plasmids originating from inpatients in Hong Kong. METHODS: Sixteen blaIMP-4-carrying plasmids identified among Enterobacteriaceae (nine Escherichia coli, four Klebsiella pneumoniae, two Citrobacter freundii and one Enterobacter cloacae) recovered from 15 patients were characterized. The isolates, collected during January 2010 to December 2013, were retrospectively investigated by plasmid sequencing, molecular and fitness studies. RESULTS: The blaIMP-4-carrying plasmids belonged to the IncN ST7 lineage (â¼50 kb). Twelve of the 16 plasmids were epidemiologically linked to seven different regions in China. Alignment of the complete plasmid sequences showed identical plasmid backbones and two highly similar resistance regions, each carrying one of two resistance genes (blaIMP-4 and qnrS1). The blaIMP-4 was detected in a class 1 integron (containing blaIMP-4 and intron Kl.pn.13) that is part of an IS6100-IS26 transposon-like structure. The nine E. coli carrying the epidemic plasmid belonged to multiple multilocus STs (six ST542, one ST131, one ST657 and one ST3177). Fitness assays performed on E. coli J53 recipients showed that the presence of the epidemic plasmid did not have a significant biological cost. CONCLUSIONS: This study identified a blaIMP-4-carrying IncN ST7 plasmid disseminated among multiple enterobacterial species originating from patients with epidemiological links to different regions in China.
Subject(s)
Cross Infection/microbiology , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae/enzymology , Plasmids/analysis , Topography, Medical , Aged , Aged, 80 and over , Cross Infection/epidemiology , Enterobacteriaceae/genetics , Enterobacteriaceae/isolation & purification , Enterobacteriaceae Infections/epidemiology , Female , Gene Transfer, Horizontal , Hong Kong/epidemiology , Humans , Infant , Male , Middle Aged , Plasmids/classification , Retrospective Studies , Sequence Alignment , Sequence Analysis, DNAABSTRACT
Increasing consumption of nitrofurantoin (NIT) for treatment of acute uncomplicated urinary tract infections (UTI) highlights the need to monitor emerging NIT resistance mechanisms. This study investigated the molecular epidemiology of the multidrug-resistant efflux gene oqxAB and its contribution to nitrofurantoin resistance by using Escherichia coli isolates originating from patients with UTI (n = 205; collected in 2004 to 2013) and food-producing animals (n = 136; collected in 2012 to 2013) in Hong Kong. The oqxAB gene was highly prevalent among NIT-intermediate (11.5% to 45.5%) and -resistant (39.2% to 65.5%) isolates but rare (0% to 1.7%) among NIT-susceptible (NIT-S) isolates. In our isolates, the oqxAB gene was associated with IS26 and was carried by plasmids of diverse replicon types. Multilocus sequence typing revealed that the clones of oqxAB-positive E. coli were diverse. The combination of oqxAB and nfsA mutations was found to be sufficient for high-level NIT resistance. Curing of oqxAB-carrying plasmids from 20 NIT-intermediate/resistant UTI isolates markedly reduced the geometric mean MIC of NIT from 168.9 µg/ml to 34.3 µg/ml. In the plasmid-cured variants, 20% (1/5) of isolates with nfsA mutations were NIT-S, while 80% (12/15) of isolates without nfsA mutations were NIT-S (P = 0.015). The presence of plasmid-based oqxAB increased the mutation prevention concentration of NIT from 128 µg/ml to 256 µg/ml and facilitated the development of clinically important levels of nitrofurantoin resistance. In conclusion, plasmid-mediated oqxAB is an important nitrofurantoin resistance mechanism. There is a great need to monitor the dissemination of this transferable multidrug-resistant efflux pump.
Subject(s)
Anti-Infective Agents, Urinary/pharmacology , Drug Resistance, Bacterial/genetics , Escherichia coli Proteins/genetics , Escherichia coli/genetics , Genes, MDR , Nitrofurantoin/pharmacology , Plasmids/metabolism , Animals , Anti-Infective Agents, Urinary/metabolism , Clone Cells , Escherichia coli/drug effects , Escherichia coli/isolation & purification , Escherichia coli/metabolism , Escherichia coli Infections/drug therapy , Escherichia coli Infections/epidemiology , Escherichia coli Infections/microbiology , Escherichia coli Proteins/metabolism , Gene Expression , Hong Kong/epidemiology , Humans , Molecular Epidemiology , Multilocus Sequence Typing , Mutation , Nitrofurantoin/metabolism , Nitroreductases/genetics , Nitroreductases/metabolism , Plasmids/chemistry , Replicon , Urinary Tract Infections/drug therapy , Urinary Tract Infections/epidemiology , Urinary Tract Infections/microbiologyABSTRACT
Of 137 Staphylococcus lugdunensis isolates collected from two nephrology centers in Hong Kong, 10 (7.3%) and 3 (2.2%) isolates had high-level and low-level mupirocin resistance, respectively. Isolates with high-level resistance contained the plasmid-mediated ileS2 gene, while isolates with low-level resistance contained the mutation V588F within the chromosomal ileS gene. All but one of the ileS2-positive isolates belong to the predominating clone HKU1. Plasmids carrying the ileS2 gene were mosaic and also cocarry multiple other resistance determinants.
Subject(s)
Drug Resistance, Multiple, Bacterial/genetics , Genes, Bacterial , Staphylococcus lugdunensis/drug effects , Staphylococcus lugdunensis/genetics , Drug Resistance, Multiple, Bacterial/drug effects , Hong Kong , Humans , Methicillin Resistance/genetics , Microbial Sensitivity Tests , Mupirocin/pharmacology , Mupirocin/therapeutic use , Mutation , Plasmids , Staphylococcus lugdunensis/isolation & purificationABSTRACT
The IncA/C plasmids are broad host-range vehicles which have been associated with wide dissemination of CMY-2 among Enterobacteriaceae of human and animal origins. Acquired metallo-ß-lactamases (MBLs) such as the IMP-type enzymes are increasingly reported in multidrug-resistant Gram-negative bacteria worldwide, particularly in Enterobacteriaceae. We described the complete sequence of the first IMP-4-encoding IncA/C2 plasmid, pIMP-PH114 (151,885 bp), from a sequence type 1 Klebsiella pneumoniae strain that was recovered from a patient who was hospitalized in the Philippines. pIMP-PH114 consists of a backbone from the IncA/C2 plasmids, with the insertion of a novel Tn21-like class 1 integron composite structure (containing the cassette array bla IMP-4-qacG-aacA4-catB3, followed by a class C ß-lactamase bla DHA-1 and the mercury resistance operon, merRTPCADE) and a sul2-floR encoding region. Phylogenetic analysis of the IncA/C repA sequences showed that pIMP-PH114 formed a subgroup with other IncA/C plasmids involved in the international spread of CMY-2, TEM-24 and NDM-1. Identical bla IMP-4 arrays have been described among different Enterobacteriaceae and Acinetobacter spp. in China, Singapore and Australia but the genetic context is different. The broad host range of IncA/C plasmids may have facilitated dissemination of the bla IMP-4 arrays among different diverse groups of bacteria.
Subject(s)
Drug Resistance, Multiple, Bacterial/genetics , Klebsiella pneumoniae/genetics , Plasmids/genetics , beta-Lactamases/genetics , Anti-Bacterial Agents/pharmacology , Gene Order , Genotype , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/drug effects , Microbial Sensitivity Tests , Phenotype , PhylogenyABSTRACT
The IncX family of plasmids has recently been expanded to include at least four subtypes, IncX1-IncX4. The revised classification provides an opportunity for improving our understanding of the sequence diversity of the IncX plasmids and the resistance genes they carried. We described the complete nucleotide sequence of a novel IncX3 plasmid, pKPC-NY79 (42,447 bp) from a sequence-type 258 Klebsiella pneumoniae strain that was isolated from a patient who was hospitalized in New York, United States. In pKPC-NY79, the plasmid scaffold and genetic load region were highly similar to homologous regions in pIncX-SHV (IncX3, JN247852) and the bla KPC carrying pKpQIL (IncFIIk, GU595196), respectively, indicating that it has possibly arisen through recombination of plasmids. The bla KPC-2 gene, as part of a transposon Tn4401a, was found within the genetic load region. The backbone of pKPC-NY79 differs from pIncX-SHV by a deletion involving the gene tandem hns-topB (encoding H-NS protein and topoisomerase III, respectively) and a putative ATPase gene. Unexpectedly, the impact of the hns-topB deletion on host fitness and plasmid stability was found to be small. In conclusion, the findings contribute to a better understanding of the plasmid platforms carrying bla KPC and of variations in the backbone of the IncX3 plasmids.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Klebsiella Infections/microbiology , Klebsiella pneumoniae/genetics , Plasmids/genetics , beta-Lactamases/genetics , Aged , Bacterial Proteins/metabolism , DNA Transposable Elements , Humans , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/isolation & purification , Male , Plasmids/metabolism , Urine/microbiology , beta-Lactamases/metabolismSubject(s)
Drug Resistance, Multiple, Bacterial/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Fosfomycin/pharmacology , Genes, Bacterial/genetics , Plasmids/genetics , Animals , Cats , Cattle , Chickens , Dogs , Escherichia coli/isolation & purification , Escherichia coli Infections , Livestock , Microbial Sensitivity Tests , Multilocus Sequence Typing , SwineSubject(s)
Bacterial Proteins/genetics , Klebsiella pneumoniae/genetics , Plasmids/genetics , beta-Lactam Resistance/genetics , beta-Lactamases/genetics , Anti-Bacterial Agents/therapeutic use , Bacterial Proteins/metabolism , China , Humans , Klebsiella Infections/drug therapy , Klebsiella Infections/microbiology , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/isolation & purification , Male , Medical Tourism , Middle Aged , Multilocus Sequence Typing , Sequence Analysis, DNA , beta-Lactamases/metabolismSubject(s)
Enterococcus faecium/drug effects , Enterococcus faecium/isolation & purification , Gram-Positive Bacterial Infections/veterinary , Livestock , Vancomycin Resistance , Animals , China , Gram-Positive Bacterial Infections/epidemiology , Gram-Positive Bacterial Infections/microbiology , PrevalenceABSTRACT
Antibiotic resistance is an emerging public health issue. Plasmids are one of the popular carriers to disseminate resistance genes among pathogens. However, the response of plasmid-carrying bacteria to antibiotic treatment and how these bacteria evolve to increase their resistance remain elusive. In this study, we conjugated plasmid pNDM-HK to E. coli J53 recipient cells and selected survivors using different concentrations of the broad spectrum antibiotic meropenem. After selection, transconjugants conferred varying minimum inhibitory concentrations with respect to carbapenems. We sequenced and compared the transcriptomes of transconjugants that exhibited distinct carbapenem susceptibilities, and found that the loss of outer membrane proteins led to antibiotic resistance. Moreover, we identified a novel mutation, G63S, in transcription factor OmpR which moderates the expression of outer membrane proteins. The loss of porins was due to incapability of phosphorylation, which is essential for porin transcription and carbapenem resistance. We also characterized other genes that are regulated by ompR in this mutant, which contributed to bacterial antibiotic resistance. Overall, our studies suggest antibiotic pressure after conjugation might be an alternative pathway to promote antimicrobial resistance.
Subject(s)
Bacterial Proteins/genetics , Carbapenems/therapeutic use , Conjugation, Genetic/physiology , Drug Resistance, Bacterial/genetics , Porins/metabolism , Trans-Activators/genetics , Bacterial Proteins/metabolism , Conjugation, Genetic/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Organisms, Genetically Modified , Permeability , Selection, Genetic , Trans-Activators/metabolismABSTRACT
Small RNAs (sRNAs) play significant roles in regulating gene expression post-transcriptionally in response to environmental changes in bacteria. In this work, we identified and characterized six novel sRNAs from an emerging multidrug-resistance (MDR) plasmid pNDM-HK, a New Delhi metallo-ß-lactamase 1 gene (blaNDM-1)-carrying IncL/M plasmid that has caused worldwide threat in recent years. These sRNAs are located at different regions of pNDM-HK, such as replication, stability, and variable regions. Moreover, one of the plasmid-encoded sRNAs (NDM-sR3) functions in an Hfq-dependent manner and possibly plays roles in the fitness of pNDM-HK carrying bacteria. In addition, we attempted to construct the phylogenetic tree based on these novel sRNAs and surprisingly, the sRNA-phylogenetic tree provided significant information about the evolutionary pathway of pNDM-HK, including possible gene acquisition and insertion from relevant plasmids. Moreover, the sRNA-phylogenetic tree can specifically cluster the IncM2 type and distinguish it from other IncL/M subtypes. In summary, this is the first study to systematically identify and characterize sRNAs from clinically-isolated MDR plasmids. We believe that these newly found sRNAs could lead to further understanding and new directions to study the evolution and dissemination of the clinically MDR bacterial plasmids.
Subject(s)
Cystitis/epidemiology , Drug Resistance, Bacterial , Escherichia coli Infections/epidemiology , Escherichia coli/classification , Escherichia coli/drug effects , Adolescent , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/pharmacology , China/epidemiology , Cystitis/microbiology , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Female , Humans , Microbial Sensitivity Tests , Middle Aged , Phenotype , Prevalence , Serotyping , Young AdultABSTRACT
The dissemination of extended-spectrum ß-lactamases (ESBLs) genes among bacteria is commonly achieved by plasmid conjugation. In the last decade, the CTX-M type enzyme was the most widespread and prevalent ESBLs in the world. In Hong Kong and mainland China, among the commonly found CTX-M-carrying plasmids were pHK01 and pHK01-like plasmids, which belong to incompatibility group FII (IncFII). In this work, we studied the physiological effect caused by the pHK01 plasmid in bacterial host Escherichia coli J53. The plasmid did not affect cell growth of the host but reduced their motility. The reduction of host motility was attributed to downregulation of genes that encode the flagellar system. We also identified several plasmid-encoded sRNAs, and showed that the overexpression of one of them, AS-traI, in the presence of pHK01 plasmid shortened the lag phase of host growth. In addition to the study of pHK01 in bacteria, we also developed a fast and incompatibility group-specific curing method using countertranscribed RNA, which could be of general usage for studying plasmid-host interaction in clinical aspects.
Subject(s)
Drug Resistance, Multiple, Bacterial/genetics , Escherichia coli/genetics , Plasmids/genetics , RNA, Bacterial/genetics , beta-Lactamases/genetics , Base Sequence , China , Escherichia coli/drug effects , Escherichia coli/growth & development , Escherichia coli Infections/microbiology , Escherichia coli Proteins/genetics , Hong Kong , Humans , Microbial Sensitivity TestsABSTRACT
Aminoglycoside resistance determinants among 188 aminoglycoside-resistant blood culture Escherichia coli isolates from a tertiary hospital in Hong Kong, from 2004 to 2010 were investigated. Overall, 91% had aac(3)-II, 12.2% had aac(6')-Ib/Ib-cr, and 5.4% had the methylase genes (rmtB, armA). Aminoglycoside-resistant isolates with aac(')-Ib/Ib-cr, rmtB, and armA often had coresistance to multiple other antibiotics.
Subject(s)
Acetyltransferases/genetics , Aminoglycosides/pharmacology , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Escherichia coli/drug effects , Escherichia coli/genetics , tRNA Methyltransferases/genetics , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Hong Kong , Humans , Prevalence , Tertiary Care CentersABSTRACT
This study investigated 248 extended-spectrum ß-lactamase-producing Escherichia coli isolates from 2012 to 2013 for hybrid blaCTX-M genes. blaCTX-M genes were detected in 228 isolates of which 14 isolates were hybrid blaCTX-M positive (6 blaCTX-M-123, 6 blaCTX-M-64, and 2 blaCTX-M-132). The 14 hybrid blaCTX-M-carrying isolates (8 from chickens, 2 each from pigs and cattle, 1 each from dog and rodent) were genetically diverse. All but 2 hybrid blaCTX-M were carried on IncI1 (5 blaCTX-M-123) and IncI2 (6 blaCTX-M-64 and one blaCTX-M-132) plasmids. Our IncI1 and IncI2 plasmids had pHNAH4-1-like and pHN1122-1-like restriction fragment length polymorphism patterns, respectively. Genetic relatedness of the plasmids to pHNAH4-1 and pHN1122-1 were confirmed by complete sequencing of 3 plasmids, pCTXM123_C0996, pCTXM64_C0967, and pCTXM132_P0421. Plasmids closely related to pHNAH4-1 and pHN1122-1 and carrying different blaCTX-M alleles have been reported from multiple geographic areas in China previously. The findings highlighted the wide dissemination of hybrid blaCTX-M variants in different parts of China.
Subject(s)
Animals, Domestic/microbiology , Escherichia coli Infections/veterinary , Escherichia coli/enzymology , Escherichia coli/genetics , Rodentia/microbiology , Urinary Tract Infections/microbiology , beta-Lactamases/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Animals , China/epidemiology , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Female , Humans , Male , Middle Aged , Plasmids/analysis , Plasmids/classification , Prevalence , Recombination, Genetic , Restriction Mapping , Sequence Analysis, DNA , Urinary Tract Infections/epidemiology , Young AdultABSTRACT
This territory-wide study investigated the occurrence of faecal carriage of extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli among wild rodents from the 18 districts in Hong Kong. Individual rectal swabs were obtained from the trapped animals and cultured in plain and selective media. A total of 965 wild rodents [148 chestnut spiny rats (Niviventer fulvescens), 326 Indo-Chinese forest rats (Rattus andamanensis), 452 brown rats (Rattus norvegicus) and 39 black rats (Rattus rattus)] were sampled. ESBL carriage was 0â% in chestnut spiny rats, 0.6â% in Indo-Chinese forest rats, 7.7â% in black rats and 13.9â% in brown rats. Among brown rats, the prevalence of ESBL carriage differed markedly by geographical location: absent in two districts, low (7-10â%) in six districts, moderate (11-19â%) in seven districts and high (21-50â%) in three districts. Nonetheless, there was no correlation between the prevalence of ESBL in brown rats and human population density in the 18 districts. CTX-M-type enzymes were detected in 92.0â% of the ESBL-producing isolates, of which 83.1â% were resistant to three or more non-ß-lactam drugs. The CTX-M producing isolates were genetically diverse but a large proportion (47.8â%) were included in six successful clones that are strongly associated with human diseases and CTX-M dissemination, viz. sequence type complex [STC]10/phylogroup A, STC23/phylogroup B1, STC38/phylogroup D, STC155/phylogroup B1, ST405/phylogroup D and ST131/phylogroup B2. In conclusion, our results show that brown rats often carry potentially zoonotic clones of CTX-M producing, multidrug-resistant E. coli. The potential for rats to be a source of CTX-M producing E. coli for humans deserves further consideration.
Subject(s)
Drug Resistance, Multiple, Bacterial , Escherichia coli Infections/veterinary , Escherichia coli/classification , Escherichia coli/enzymology , Molecular Typing , Rodentia/microbiology , beta-Lactamases/metabolism , Animals , Carrier State/microbiology , Carrier State/veterinary , Escherichia coli/genetics , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Feces/microbiology , Female , Genetic Variation , Genotype , Hong Kong , Male , RatsABSTRACT
Previous work on the subclones within Escherichia coli ST131 predominantly involved isolates from Western countries. This study assessed the prevalence and antimicrobial resistance attributed to this clonal group. A total of 340 consecutive, non-duplicated urinary E. coli isolates originating from four clinical laboratories in Hong Kong in 2013 were tested. ST131 prevalence among the total isolates was 18.5â% (63/340) and was higher among inpatient isolates (23.0â%) than outpatient isolates (11.8â%, P<0.001), and higher among isolates from patients aged ≥65 years than from patients aged 18-50 years and 51-64 years (25.4 vs 3.4 and 4.0â%, respectively, P<0.001). Of the 63 ST131 isolates, 43 (68.3â%) isolates belonged to the H30 subclone, whereas the remaining isolates belonged to H41 (nâ=â17), H54 (nâ=â2) and H22 (nâ=â1). All H30 isolates were ciprofloxacin-resistant, of which 18.6â% (8/43) belonged to the H30-Rx subclone. Twenty-six (41.3â%) ST131 isolates were ESBL-producers, of which 19 had blaCTX-M-14 (12 non-H30-Rx, two H30-Rx and five H41), six had blaCTX-M-15 (five non-H30-Rx and one H30-Rx) and one was blaCTX-M-negative (H30). In conclusion, ST131 accounts for a large share of the antimicrobial-resistant E. coli isolates from geriatric patients. Unlike previous reports, ESBL-producing ST131 strains mainly belonged to non-H30-Rx rather than the H30-Rx subclone, with blaCTX-M-14 as the dominant enzyme type.
Subject(s)
Anti-Infective Agents/pharmacology , Drug Resistance, Bacterial/genetics , Escherichia coli Infections/epidemiology , Escherichia coli/isolation & purification , Urinary Tract Infections/epidemiology , Adolescent , Adult , Aged , Alleles , Bacterial Typing Techniques , DNA Primers/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli Infections/microbiology , Escherichia coli Proteins/genetics , Female , Geriatrics , Humans , Male , Middle Aged , Prevalence , Species Specificity , Urinary Tract/microbiology , Urinary Tract Infections/microbiology , Young Adult , beta-Lactamases/geneticsABSTRACT
This study investigated the prevalence of IncX plasmid subtypes in commensal and pathogenic Escherichia coli isolates and the biological features of the IncX4 subtype. Two hundred and twenty-five E. coli isolates from multiple sources (47 chickens, 41 pigs, 30 cattle and 107 humans) obtained during the period 2006-2012 were tested for the presence of IncX1 to IncX5. Overall, the prevalence of IncX plasmids in chicken, pig, cattle and human isolates were 21.2â% (10/47), 19.5â% (8/41), 3.3â% (1/30) and 4.8â% (5/107), respectively. IncX4 was the most common subtype, followed by IncX1 and IncX3, while no IncX2 or IncX5 were found. Seven out of 16 (43.8â%) IncX4 plasmids were found to carry blaCTX-M genes and six of them originating from different host sources (four chickens, one pig and one human) had identical or highly similar RFLP patterns. Three IncX4 plasmids carrying blaCTX-M from different host sources were investigated further. It was found that the IncX4 plasmids had little effect on bacterial host growth parameters after their introduction to J53 recipients. Conjugation experiments demonstrated that the IncX4 plasmids could be efficiently transferred at 30-42 °C at rates which were generally 10(2)-10(5)-fold higher than those for the epidemic IncFII plasmid carrying blaCTX-M (pHK01). In conclusion, the IncX plasmids are more common than previously recognized. The efficient transfer of IncX4 plasmid at different temperatures and the lack of fitness burden on bacterial hosts highlight the ability of this plasmid replicon to be an important vehicle for dissemination of antimicrobial resistance.
Subject(s)
Escherichia coli Proteins/metabolism , Escherichia coli/metabolism , Livestock , beta-Lactamases/metabolism , Animals , Escherichia coli/genetics , Escherichia coli Infections/microbiology , Escherichia coli Infections/veterinary , Escherichia coli Proteins/genetics , Gene Expression Regulation, Bacterial/physiology , Humans , Molecular Sequence Data , Plasmids , beta-Lactamases/geneticsABSTRACT
A total of 1878 non-duplicate clinical Escherichia coli isolates (comprising 1711 urinary isolates and 167 blood-culture isolates), which were collected from multiple centres in Hong Kong during 1996-2008, were used to investigate the prevalence and molecular epidemiology of plasmid-mediated fosfomycin (fos) resistance genes. Eighteen of the 1878 clinical E. coli isolates were fosfomycin resistant, of which six were fosA3 positive and two were positive for another fosA variant (designated fosKP96). No isolates had the fosC2 gene. The clones of the eight isolates were diverse: sequence type (ST) 95 (nâ=â2), ST118 (nâ=â1), ST131 (nâ=â1), ST617 (nâ=â1), ST648 (nâ=â1), ST1488 (nâ=â1) and ST2847 (nâ=â1). In the isolates, fosA3 and blaCTX-M genes were co-harboured on conjugative plasmids with F2:A-:B- (nâ=â2), N (nâ=â1), F-:A-:B1 and N (nâ=â1) and untypable (nâ=â2) replicons. Both fosKP96-carrying plasmids belonged to replicon N. RFLP analysis showed that the two F2:A-:B- plasmids carrying fosA3 and blaCTX-M-3 genes shared the same pattern. Complete sequencing of one of the two F2:A-:B- plasmids, pFOS-HK151325 (69â768 bp) demonstrated it to be >99â% identical to the previously sequenced plasmid pHK23a originating from a pig E. coli isolate in the same region. This study demonstrated the dissemination of fosA3 genes in diverse E. coli clones on multiple blaCTX-M-carrying plasmid types, of which F2:A-:B- plasmids closely related to pHK23a were shared by isolates from human and animal sources.