Dissemination of clonally related Escherichia coli strains expressing extended-spectrum [beta]-lactamase CTX-M-15.
We analyzed 43 CTX-M-15--producing Escherichia coli isolates and 6 plasmids encoding the [bla.sub.CTX-M-15] gene from Canada, India, Kuwait, France, Switzerland, Portugal, and Spain. Most isolates belonged to phylogroups B2 (50%) and D (25%). An EC-B2 strain of clonal complex sequence type (ST) 131 was detected in all countries; other B2 isolates corresponded to ST28, ST405, ST354, and ST695 from specific areas. EC-D strains were clonally unrelated but isolates from 3 countries belonged to ST405. All CTX-M-15 plasmids corresponded to IncFII group with overrepresentation of 3 Hpal-digested plasmid DNA profiles (A, B and C; 85-120kb, similarity [greater than or equal to]70%). Plasmid A was detected in EC-B2 strains (ST131, ST354, or ST405), plasmid C was detected in B2 and D strains, and plasmid B was confined to worldwide-disseminated ST131. Most plasmids contained [bla.sub.OXA-1], aac(6')-lb-cr, and [bla.sub.TEM TEM
1. transmission electron microscope.
3. transmissible encephalopathy of mink. .1]. Worldwide dissemination of CTX-M-15 seems to be determined by clonal complexes ST131 and ST405 and multidrug-resistant IncFII plasmids.
Plasmid-mediated CTX-M type expanded-spectrum [beta]-lactamases (ESBLs), which have been extensively reported for the past 10 years, are detected mostly in community-acquired pathogens and are associated mainly with Escherichia coli. These [beta]-lactamases compromise the efficacy of all [beta]-lactams, except carbapenems and cephamycins, and are associated with many non-[beta]-lactam resistance markers because of their locations on plasmids. Therefore, they may constitute a real threat for treating community-acquired E. coli-mediated urinary tract infections (1,2).
Different variants of CTX-M ESBLs are grouped in 5 clusters, although their distribution varies greatly depending on the geographic area (www.lahey.org/studies/webt. htm). CTX-M-15, which was first detected in isolates from India in 2001 (3), is now recognized as the most widely distributed CTX-M enzyme. It is derived from CTX-M3 by 1 amino acid substitution at position 240 (Asp-240 [right arrow] Gly), which apparently confers an increased catalytic activity to ceftazidime (4). Clonal outbreaks of CTX-M15-producing Enterobacteriaceae have been reported in France, Italy, Spain, Portugal, Austria, Norway, the United Kingdom, Tunisia, South Korea, and Canada, and E. coli is the most frequently involved species. Within E. coli, CTXM-15-producing strains of the B2 phylogenetic group are commonly found and frequently harbor multidrug resistance and virulence determinants (5-18).
Plasmids encoding [bla.sub.CTX-M-15] have been isolated from clinical isolates in France, Spain, Portugal, the United Kingdom, Canada, India, Pakistan, South Korea, Taiwan, the United Arabic Emirates, and Honduras (5-8, 10,11,15,19,20). Plasmid characterization, which has only been accomplished for those plasmids from Canada, France, Spain, and the United Kingdom, classified most of them as members of incompatibility group FII FII Federated Investors Inc. (Pittsburgh, PA)
FII Foreign Institutional Investor
FII Falling Into Infinity (Dream Theater album)
FII Fundación Instituto de Ingeniería (5,7,8,17,19).
Lack of detailed studies on isolates expressing particular CTX-Ms from different geographic areas has precluded identification of factors involved in recent and worldwide spread of specific CTX-M variants. In this article, through analysis of the population biology of CTX-M-15--producing isolates from 7 countries and characterization of their genetic elements, we provide a comprehensive picture of elements involved in international spread of a particularly widespread mechanism of antimicrobial drug resistance.
Materials and Methods
Bacterial Strains, Production of ESBL ESBL Extended Spectrum Beta Lactamase
ESBL East Staffordshire Badminton League (UK) , and Susceptibility Testing
We studied 43 CTX-M-15--producing E. coli clinical isolates from France (n = 17), Kuwait (n = 9), Switzerland (n = 7), Canada (n = 4), Portugal (n = 3) and Spain (n = 3), and 6 CTX-M-15 plasmids from India (3), all obtained from 2000 through 2006. These strains and plasmids were considered representative of these areas because they either caused outbreaks or were the first isolates recovered in those countries (3,11,16,19,21-23). Samples were isolated from urine (n = 33/43, 77%), wounds (n = 4/43, 9.%), respiratory tract infections (n = 3/43, 7%) and other sites (1 from feces, 1 from an intravenous catheter, and 1 from blood) in hospitalized patients. ESBL production was confirmed by a standard double-disk synergy test, and bla genes were characterized by PCR PCR polymerase chain reaction.
polymerase chain reaction
Polymerase chain reaction (PCR) and additional sequencing as described (19). Susceptibility patterns to 13 non-[beta]-lactam antimicrobial drugs were determined by the standard disk diffusion method following published standards (24). Strains with intermediate susceptibility were considered resistant.
Clonal relationships were established by pulsed-field gel electrophoresis (PFGE PFGE Pulsed-Field Gel Electrophoresis ) of XbaI-digested genomic DNA (New England Biolabs New England Biolabs (NEB) produces and supplies reagents for the life science industry. NEB offers a large selection of recombinant and native enzymes for genomic research. It also offers products in the areas related to proteomics and drug discovery. , Ipswich, MA, USA) as described (25). Assignment of E. coli phylogenetic groups was conducted by using a multiplex PCR assay described by Clermont et al. (26). All E. coli isolates belonging to phylogroups B2 and D were characterized by multilocus sequence typing Multilocus sequence typing (MLST) is a technique in molecular biology for the typing of multiple loci. The procedure characterizes isolates of bacterial species using the DNA sequences of internal fragments of multiple (usually seven) housekeeping genes. (MLST MLST Multi Locus Sequence Typing
MLST Medical Logistics Support Team
MLST Mini Losi Super Truck (1/18th scale radio control vehicle) ) using the standard 7 housekeeping loci (www.mlst.net). All fumC sequences from E. coli isolates belonging to phylogroup D were analyzed for a C288T single nucleotide polymorphism Noun 1. single nucleotide polymorphism - (genetics) genetic variation in a DNA sequence that occurs when a single nucleotide in a genome is altered; SNPs are usually considered to be point mutations that have been evolutionarily successful enough to recur in a . This polymorphism is specific for a globally disseminated E. coli strain arbitrarily designated as E. coli clonal group A (CgA) that is associated with community-acquired urinary tract infections (27,28).
Transferability and Location of [bla.sub.CTX-M-15]
Transferability was tested by broth and filter mating assays using E. coli K12 strain BM21R (resistant to nalidixic acid and rifampin rifampin (rĭfăm`pĭn), antibiotic used in the treatment of tuberculosis. It is also used to eliminate the meningococcus microorganism from carriers and to treat leprosy, or Hansen's disease. , positive for lactose fermentation, and free of plasmids) as recipient at a 1:2 donor: recipient ratio. Transconjugants were selected on Luria-Bertani agar plates containing cefotaxime (1 mg/L) and rifampin (100 mg/L) and incubated at 37[degrees]C for 24-48 h. Transformation was performed for a subset of isolates by using conditions reported (3). Chromosomal or plasmid location of [bla.sub.CTX-M-15] genes was assessed by hybridization hybridization /hy·brid·iza·tion/ (hi?brid-i-za´shun)
1. crossbreeding; the act or process of producing hybrids.
2. molecular hybridization
3. of I-CeuI-digested genomic DNA with [bla.sub.CTX-M-15] and 16S rDNA probes and electrophoresis (5-25 s for 23 h and 60-120 s for 10 hat 14[degrees]C and 6 V/[cm.sup.2]) (25). Transfer and hybridization were performed by using standard procedures. Labeling and detection were conducted by using enhanced chemiluminescence chemiluminescence /chemi·lu·mi·nes·cence/ (kem?i-loo?mi-nes´ens) luminescence produced by direct transformation of chemical energy into light energy. (Amersham Life Sciences, Uppsala, Sweden) following manufacturer's instructions.
Plasmid DNA was obtained by using different midiprep plasmid purifcation kits (QIAGEN, Hilden, Germany, and Marlingen Biosciences, Ijamsville, MD, USA). Plasmids were classified according to their incompatibility group by a PCR-based replicon-typing scheme (29). Determination of plasmid size and confirmation of replicon rep·li·con
A genetic element that undergoes replication as an autonomous unit. content was established for transconjugants (or wild-type strains in the absence of transfer) by hybridization of S1 nuclease--digested genomic DNA with probes specific for [bla.sub.CTX-M-15] and for different F replicons (FII, FIA FIA
feline infectious anemia. , FIB), which were obtained by PCR as described (19). Relationships among plasmids were determined by comparison of EcoRI and HpaI digested DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. patterns and comparison of repFII sequences. Genescan software (Applied Biosystems, Foster City, CA, USA) was used for collection of gel images. Data of a subset of representative patterns were exported into Fingerprinting II Informatix version 3.0 software (Bio-Rad Laboratories, Hercules, CA, USA) for further interpretation. Cluster analysis was conducted by using the unweighted pair group method with arithmetic averages (optimization 0.5%, tolerance 1.00%).
Presence of genes previously associated with plasmids encoding CTX-M-15 as [bla.sub.OXA-1], [bla.sub.TEM-1], and aac(6')-Ib-cr was screened by PCR by using primers [bla.sub.OXA-1] (oxal FW: 5'-TTT TCT TCT The Capital Times (Madison, WI newspaper)
TCT Transcatheter Cardiovascular Therapeutics
TCT The Coroner's Toolkit
TCT Trans Canada Trail
TCT Tcl Core Team
TCT Tsukuba College of Technology (Japan) GTT GTT,
n See test, glucose tolerance.
GTT Glucose tolerance test, see there GTT TGG TGG The Great Gatsby (novel F. Scott Fitzgerald; movie)
TGG Kuala Terengganu, Malaysia - Sultan Mahmood (Airport Code)
TGG Temporary Geographic Grid
TGG Third Generation Gyro
TGG Triple Graph Grammar GTT TT-3' and oxal RV: 5'TTT CTT CTT Correios (Portuguese Postal Service)
CTT Certified Technical Trainer
CTT Charity Technology Trust
CTT Cholesterol Treatment Trialists' (collaboration)
CTT Common Task Training GGC GGC Girl Guides of Canada
GGC Greenwood Genetic Center (South Carolina)
GGC Gwasanaeth Gwaed Cymru (Welsh Blood Service)
GGC Generalized Goppa Code
GGC Grosvenor Gallery Company TTT "Thought that too." See digispeak. TAT GCT (programming, tool) GCT - A test-coverage tool by Brian Marick <email@example.com>, based on GNU C. Version 1.4 was ported to Sun-3, Sun-4, RS/6000, 68000, 88000, HP-PA, IBM 3090, Ultrix, Convex, SCO but not Linux, Solaris, or Microsoft Windows. TG-3'), [bla.sub.TEM-1] (TEM-F: 5'-ATG AGT AGT antiglobulin test. ATT ATT
ammonia tolerance test. CAA Caa
See CCC. CAT TTC TTC Trying To Conceive
TTC Toronto Transit Commission
TTC Trans Texas Corridor
TTC Toutes Taxes Comprises (French)
TTC Trident Technical College (North Charleston, SC)
TTC Temporary Traffic Control CG-3' and TEM-R: 5'-CTG ACA ACA - Application Control Architecture GTT ACC See adaptive cruise control. AAT Alpha-1-antitrypsin (AAT)
A blood component that breaks down infection-fighting enzymes such as elastase.
Mentioned in: Chronic Obstructive Lung Disease GCT TA-3'), and aac(6')Ib-cr (aac-cr-F: 5'-TTG CGA TGC TGC The Golf Channel
TGC The Game Creators (forum)
TGC Trading Card Game
TGC Time-Gain Compensation
TGC The Gungan Council
TGC The Golden Compass (Phillip Pullman book)
TGC Take Good Care TCT ATG ATG antithymocyte globulin.
lymphocyte immune globulin (antithymocyte globulin equine, ATG, ATG equine, LIG)
Pharmacologic class: Immunoglobulin
Therapeutic class: Immunosuppressant AGT GG-3' and aac-cr-R: 5'-GCG TGT TGT Target
TGT Ticket Granting Ticket (Windows 2000 Kerberos security)
TGT Target Corp (stock symbol)
TGT Turbine Gas Temperature
TGT TDRSS Ground Terminal
TGT Tank Gunnery Trainer
TGT Target Tracker TCG (Trusted Computing Group, Beaverton, OR, www.trustedcomputinggroup.org) The successor to the Trusted Computer Platform Alliance (TCPA), announced in 2003 by founding members AMD, HP, IBM, Intel and Microsoft. CTC CTC - Cornell Theory Center GAA GAA Goals Against Average (Hockey)
GAA Gaelic Athletic Association
GAA Gravure Association of America (Rochester, NY)
GAA German Agro Action
GAA Global Aquaculture Alliance
GAA Gay Activists Alliance TGC C-3') (11,19,30). Additional sequencing was necessary to identify the corresponding genes.
Most CTX-M-15--producing E. coli isolates belonged to phylogroups B2 (50%) and D (25%), which are known to be associated with the hospital setting and extraintestinal pathogenic E. coli. Phylogroups A (18%) and B1 (7%), which are associated with animal or human commensal commensal /com·men·sal/ (kom-men´sil)
1. living on or within another organism, and deriving benefit without harming or benefiting the host.
2. a parasite that causes no harm to the host. strains, were less frequently represented. All isolates of phylogroups B2, A, and D corresponded to subgroups B23, [A.sub.1], and [D.sub.1], respectively, which are the most common ones within each phylogenetic group (31). The 43 clinical isolates were classified into 32 PFGE types ([B2.sub.3], 13; [D.sub.1], 10; [A.sub.l], 6; and B1, 3). Among [B2.sub.3] strains, 10 PFGE types (18 isolates from France, Canada, Spain, Portugal, Kuwait, and Switzerland) were possibly related according to criteria of Tenover et al. (32) (difference [less than or equal to] 6 bands, [greater than or equal to] 80% similarity) and were assigned to the sequence type (ST) ST131. The 4 unrelated B2 strains were classified within ST695 (1 from France), ST28 (1 from Switzerland), ST354 (1 from Portugal and Spain) and ST405 (1 from Portugal). All isolates of phylogroup [D.sub.1] were clonally unrelated by PFGE (difference [greater than or equal to] 6 bands), although MLST studies indicated that 4 PFGE types (5 isolates) from Kuwait, Switzerland, and Spain corresponded to ST405. The fumC sequences of the remaining 6 E. coli D strains were highly diverse (alleles 4, 13, 26, 88, and 132). None of the strains had the C288T single nucleotide polymorphism specific for E. coli strain CgA (28). All 3 B1 isolates were found in France. Among B2 E. coli isolates, all but 4 were isolated from urine and all but 2 belonged to ST131. These strains correspond to 2 isolates recovered from wounds and identified as ST28 and ST354 and 2 ST131 isolates from respiratory and fecal samples, respectively.
CTX-M-15 clinical strains were considered resistant to different antimicrobial drugs: amoxicillin-clavulanate (98%), tobramycin tobramycin /to·bra·my·cin/ (to?brah-mi´sin) an aminoglycoside antibiotic derived from a complex produced by Streptomyces tenebrarius, (89%), kanamycin kanamycin /kan·a·my·cin/ (kan?ah-mi´sin) an aminoglycoside antibiotic derived from Streptomyces kanamyceticus, effective against aerobic gram-negative bacilli and some gram-positive bacteria, including mycobacteria; used as the (87%), tetracycline tetracycline (tĕ'trəsī`klēn), any of a group of antibiotics produced by bacteria of the genus Streptomyces. They are effective against a wide range of Gram positive and Gram negative bacteria, interfering with protein (84%), gentamicin gentamicin /gen·ta·mi·cin/ (jen?tah-mi´sin) an aminoglycoside antibiotic complex isolated from bacteria of the genus Micromonospora, (82%), nalidixic acid (74%), streptomycin streptomycin (strĕp'tōmī`sĭn), antibiotic produced by soil bacteria of the genus Streptomyces and active against both gram-positive and gram-negative bacteria (see Gram's stain), including species resistant to other (68%), sulfonamides Sulfonamides Definition
Sulfonamides are medicines that prevent the growth of bacteria in the body.
Sulfonamides are used to treat many kinds of infections caused by bacteria and certain other microorganisms. (61%), ciprofloxacin (61%), trimethoprim trimethoprim /tri·meth·o·prim/ (-meth´o-prim) an antibacterial closely related to pyrimethamine; almost always used in combination with a sulfonamide, primarily for the treatment of urinary tract infections. (58%), chloramphenicol chloramphenicol (klōr'ămfĕn`əkŏl'), antibiotic effective against a wide range of gram-negative and gram-positive bacteria (see Gram's stain). It was originally isolated from a species of Streptomyces bacteria. (21%), nitrofurantoin nitrofurantoin /ni·tro·fu·ran·to·in/ (-fu-ran´to-in) an antibacterial effective against many gram-negative and gram-positive organisms; used in urinary tract infections.
n. (12%), and amikacin (11%). All CTX-M-15 transconjugants expressed resistance to aminoglycosides, tetracycline, or trimethoprim. All but 2 strains contained [bla.sub.OXA-1] and aac(6')-Ib-cr; 1 contained only aac(6')-Ib-cr, and 1 contained [bla.sub.OXA-1] and aacA4, which confers reduced susceptibility to amikacin and kanamycin.
Location and Transferability of [bla.sub.CTX-M-15]
The [bla.sub.CTX-M-15] gene was located on plasmids in all but 6 strains and was positively transferred by conjugation conjugation, in genetics
conjugation, in genetics: see recombination.
conjugation, in grammar
conjugation: see inflection. or transformation in 37% of the strains tested. In 8 clinical isolates corresponding to 7 PFGE types, the probe for [bla.sub.CTX-M-15] hybridized in chromosomal bands (2 belonging to [B2.sub.3] ST131, 2 to [D.sub.1], 1 to [D.sub.1] ST405, and 1 to [A.sub.1]). In 2 other strains, the [bla.sub.CTX-M-15] probe hybridized both with plasmid and chromosomal bands (1 strain from D ST405 and 1 from phylogroup B1).
Plasmids Encoding CTX-M-15
Plasmids positive for the [bla.sub.CTX-M-15] gene showed variable sizes (85-160 kb), belonged to the narrow host range incompatibility group IncF, and had replicon FII alone or in association with the FIA or FIB replicons (online Appendix Table, available from www.cdc.gov/EID/content/14/2/ 195-appT.htm). Many restriction fragment length polymorphism restriction fragment length polymorphism
n. Abbr. RFLP
Intraspecies variations in the length of DNA fragments generated by the action of restriction enzymes and caused by mutations that alter the sites at which these enzymes act, changing (RFLP RFLP
restriction fragment length polymorphism
restriction fragment length polymorphism.
RFLP ) patterns were observed, with overrepresentation of 3 profiles corresponding to 3 plasmids arbitrarily designated as plasmid A (85 kb), plasmid B (120 kb), and plasmid C (85 kb). Plasmid A, which was isolated from B2 E. coli strains from 4 countries (India, France, Portugal, and Spain), was associated with different STs (ST131, ST354, or ST405). Plasmid C was also detected in clonally unrelated E. coli of phylogroups B2 and D from Switzerland, Canada and France. Plasmid B, which was only associated with E. coli ST131, was widely disseminated in all countries studied. Sequence analysis of the replicons showed 4 repFII types: repFII(1), which was identical to that of plasmids R100, NR1, or pC15-1a, and was the most represented and identified in 23 plasmids; repFII(2), which had 99%-100% homology with plasmid pRSB 107 (GenBank accession no. AJ851089), was identified in 6 plasmids; and repFII(3) and repFII(4), which were detected in 2 and 7 plasmids, respectively, and showed >93% homology with repFII(1). All repFIA and repFIB sequences were 99% and 100% homologous, respectively, with that of pRSB107 (GenBank accession no. AJ851089).
Computer analysis of representative RFLP patterns and repFII sequences grouped CTX-M-15 plasmids within 3 major clusters with similarity >70%. Cluster I comprises most plasmids, including plasmids A and B, most containing repFII(1) and showing variable replicon content. Cluster II comprised only plasmid C derivatives showing slightly different repFII sequences, and cluster III included 2 plasmids carrying repFII(2), FIA, and FIB replicons (Figure).
In the 8 strains with chromosomal location of [bla.sub.CTX-M-15], repFII plasmids were identified but these plasmids were negative for the [bla.sub.CTX-M-15] gene. Several strains that were also positive for additional plasmids and negative for the [bla.sub.CTX-M-15] gene were assigned to different incompatibility groups or were untypeable by the PCR-based replicon typing scheme used.
Our study indicates that current worldwide spread of the [bla.sub.CTX-M-15] gene is driven mainly by 2 epidemic E. coli strains belonging to phylogroups B2 (ST131) and D (ST405) and by its location on IncF plasmids harboring multiple antimicrobial drug-resistance determinants, including the recently described aac(6')-Ib-cr gene. The presence of [bla.sub.CTX-M-15] has previously been associated with E. coli strains of phylogroups B2 and D, and in some instances, with specific PFGE types (9-12,16). We detected an emerging and globally disseminated CTX-M-15 phylogroup B2 E. coli strain corresponding to the ST 131 that was responsible for clonal outbreaks in Canada, France, Spain, and Portugal (11,14,16,23). Other CTX-M-15 B2 strains belong to clonal complexes ST695, ST405, ST354, or ST28, which have previously been detected in different geographic areas among isolates that do not express CTX-M-15 (online Appendix Figure, available from www.cdc.gov/EID/content/14/2/195-appG.htm).
Globally disseminated E. coli strains associated with acute, uncomplicated, community-acquired cystitis and pyelonephritis pyelonephritis: see nephritis.
Infection (usually bacterial) and inflammation of kidney tissue and the renal pelvis. Acute pyelonephritis is usually localized and may have no apparent cause. , designated in community patients as clone CgA (ST69), have only been occasionally associated with CTX-M-15 production in Canada (16,27,28). Although the isolates in our study do not belong to clone CgA, they were isolated mainly from urine samples, and an association of ST131 E. coli isolates with urinary tract infections might be inferred. Although most CTX-M-15 isolates studied were recovered from hospitalized patients, these microorganisms are now widely spread in the community setting, including long-term care facilities in the countries from which isolates included in this study originated (2,5,14,33). Our study has increased knowledge of the number of epidemic E. coli clonal complexes causing urinary tract infections.
All plasmids carrying [bla.sub.CTX-M-15] included in this study corresponded to incompatibility group F, and all had the FII replicon, which was assorted mainly in multireplicon plasmids with additional replicons of the FIA and FIB types. Association of the [bla.sub.CTX-M-15] gene with IncFII replicons has been described in studies conducted in Canada, France, Spain, and the United Kingdom (5,7,8,17,19). Although we observed intercontinental dissemination of 3 major IncFII plasmid scaffolds (A, B, and C) carrying [bla.sub.CTX-M-15], similarity >70% among all variants studied and presence of genes also found in pC15-1a, a CTX-M-15 plasmid (GenBank accession no. AY458016) that has a 28.4-kb multidrug resistance region containing [bla.sub.TEM-1], [bla.sub.OXA-1], the aac(6')-Ib-cr gene (aminoglycoside aminoglycoside /ami·no·gly·co·side/ (-gli´ko-sid) any of a group of antibacterial antibiotics (e.g., streptomycin, gentamicin) derived from various species of Streptomyces 6'-N-acetyltransferase type Ib-cr variant responsible for reduced susceptibility to both aminoglycosides and certain fluoroquinolones), and genetic determinants coding for resistance to tetracycline and aminoglycosides (5,30), suggest a common origin or a common particular plasmid scaffold involved in the dissemination of CTX-M-15.
Because IncF plasmids are a heterogeneous and largely diffused family of plasmids in E. coli, they could acquire the [bla.sub.CTX-M-15] gene. IncF plasmids negative for the [bla.sub.CTX-M-15] gene in strains with this gene at a chromosomal location also suggest dynamic horizontal exchanges between the chromosome and resident plasmids. Extensive recombination events among IncF plasmids are frequent and may have contributed to their apparent high diversity (variable rep content, plasmid size, transferability, antimicrobial drug-resistance genes), driving their evolution and enabling them to persist in diverse E. coli populations (34,35). Such recombination events among plasmids of the same incompatibility group within the same cell occur frequently (34,35). This hypothesis is supported by the results of Lavollay et al. (17), who described mosaicism in a CTXM-15 plasmid isolated in France that contained genes from 2 different IncFII plasmids, pC15-1a and pRSB107 (from IncFII plasmids first isolated from persons in Canada and activated sludge bacteria from a wastewater treatment plant Wastewater treatment plant also called wastewater treatment works
Spread and maintenance of conjugative plasmids across bacterial populations have been intensively studied from a theoretical point of view, but data from natural populations are scarce (34,37,38). Recovery of related plasmids from clonally unrelated B2 strains might reflect efficient transfer of these elements among different B2 E. coli populations. Sharing the same environment, successive immigrant B2 strains might sweep through the population, enabling plasmid hitchhiking Hitchhiking (also known as lifting, thumbing, hitching, autostop or thumbing up a ride) is a means of transportation that is gained by asking people (usually strangers) for a ride in their automobile to travel a distance that may either be a short or long distance. at a high frequency in each selective sweep. However, we lack detailed information on the specificity and stability of different plasmid groups in specific hosts. An evolutionary convergent relationship among B2 genetic background and IncFII plasmids cannot be ruled out and should be studied because it might explain successful dissemination of CTX-M-15 plasmids within this E. coli lineage. In addition, our study is one of the few that have identified [bla.sub.ESBL] genes in the chromosome, which might respond either to plasmid integration or transposition transposition /trans·po·si·tion/ (trans?po-zish´un)
1. displacement of a viscus to the opposite side.
2. driven by ISEcpl located upstream from the [bla.sub.CTX-M-15] gene (25,39,40).
In conclusion, worldwide dissemination of [bla.sub.CTX-M-15] is driven by B2 or D E. coli clones associated mainly with urinary tract infections or IncFII plasmids containing a multiple antimicrobial drug-resistance platform that contributes to spread of CTX-M-15. Further studies to test the stability/variability and fitness of particular plasmids among different bacterial hosts will be relevant in developing additional strategies to control dissemination of antimicrobial drug resistance.
We thank C. de Champs, A. Wenger, and V. Rotimi for providing some of the strains used in this study.
This work was partially supported by research grants from the Ministerio de Ciencia y Tecnologia of Spain (grant SAF SAF Safety
SAF Society of American Foresters
SAF Society of American Florists
SAF Secretary of the Air Force
SAF Second Amendment Foundation
SAF Singapore Armed Forces
SAF Students for Academic Freedom
SAF Store And Forward 2003-09285) to T.M.C. and the European Commission (grants LSHM-CT-2003-503335 and LSHM-CT-2005-018705). A.N. was supported by fellowships from the Ministerio de Ciencia y Tecnologia of Spain (SAF 2003-09285) and the Centro de Investigacion Biomedica en Red de Epidemiologia y Salud Publica.
Dr Coque is a senior scientist in the Microbiology Department at the Hospital Universitario Ramon y Cajal Ra·mòn y Ca·jal , Santiago 1852-1934.
Spanish histologist. He shared a 1906 Nobel Prize for research on the nervous system. in Madrid. Her research interests include the molecular epidemiology, ecology, and evolution of antimicrobial drug--resistant Enterobacteriaceae and Enterococcus enterococcus /en·tero·coc·cus/ (en?ter-o-kok´us) pl. enterococ´ci an organism belonging to the genus Enterococcus.
Enterococcus /En·tero·coc·cus/ ( .
(1.) Canton R, Coque TM. The CTX-M beta-lactamase pandemic. Curr Opin Microbiol. 2006;9:466-75.
(2.) Livermore DM, Canton R, Gniadkowski M, Nordmann P, Rossolini GM, Arlet G, et al. CTX-M: changing the face of ESBLs in Europe. J Antimicrob Chemother. 2007;59:165-74.
(3.) Karim A, Poirel L, Nagarajan S, Nordmann P. Plasmid-mediated extended-spectrum beta-lactamase (CTX-M-3 like) from India and gene association with insertion sequence ISEcpl. FEMS Microbiol Lett. 2001;201:237-41.
(4.) Poirel L, Gniadkowski M, Nordmann P. Biochemical analysis of the ceftazidime-hydrolysing extended-spectrum beta-lactamase CTXM-15 and of its structurally related beta-lactamase CTX-M-3. J Antimicrob Chemother. 2002;50:1031-4.
(5.) Boyd DA, Tyler S, Christianson S, McGeer A, Muller MP, Willey BM, et al. Complete nucleotide sequence of a 92-kilobase plasmid harboring the CTX-M-15 extended-spectrum beta-lactamase involved in an outbreak in long-term-care facilities in Toronto, Canada. Antimicrob Agents Chemother. 2004;48:3758-64.
(6.) Eisner A, Fagan EJ, Feierl G, Kessler HH, Marth E, Livermore DM, et al. Emergence of Enterobacteriaceae isolates producing CTX-M extended-spectrum beta-lactamase in Austria. Antimicrob Agents Chemother. 2006;50:785-7.
(7.) Hopkins KL, Liebana E, Villa L, Batchelor M, Threlfall EJ, Carattoli A. Replicon typing of plasmids carrying CTX-M or CMY (Cyan Magenta Yellow) The color space used for printing. In theory, equal amounts of all three colors produce black. In practice, a separate black ink is required for quality printing. See CMYK. beta-lactamases circulating among Salmonella and Escherichia coli isolates. Antimicrob Agents Chemother. 2006;50:3203-6.
(8.) Karisik E, Ellington MJ, Pike R, Warren RE, Livermore DM, Woodford N. Molecular characterization of plasmids encoding CTX-M-15 beta-lactamases from Escherichia coli strains in the United Kingdom. J Antimierob Chemother. 2006;58:665-8.
(9.) Lavigne JP, Marchandin H, Delmas J, Moreau J, Bouziges N, Lecaillon E, et al. CTX-M-producing Escherichia coli in French hospitals: prevalence, molecular epidemiology, and risk factors. J Clin Microbiol. 2006;45:620-6.
(10.) Leflon-Guibout V, Jurand C, Bonacorsi S, Espinasse F, Guelfi MC, Duportail F, et al. Emergence and spread of three clonally related virulent isolates of CTX-M-15--producing Escherichia coli with variable resistance to aminoglycosides and tetracycline in a French geriatric hospital. Antimicrob Agents Chemother. 2004;48: 3736-42.
(11.) Machado E, Coque TM, Canton R, Baquero F, Sousa JC, Peixe L. Dissemination of Enterobacteriaceae harboring [bla.sub.CTX-M-15], [bla.sub.OXA-1], [bla.sub.TEM-1] and aac(6')-Ib-cr gene in Portugal. Antimicrob Agents Chemother. 2006;50:3220-1.
(12.) Mamlouk K, Boutiba-Ben Boubaker I, Gautier V, Vimont S, Picard B, Ben Redjeb S, et al. Emergence and outbreaks of CTX-M betalactamase--producing Escherichia coli and Klebsiella pneumoniae strains in a Tunisian hospital. J Clin Microbiol. 2006;44:4049-56.
(13.) Mugnaioli C, Luzzaro F, de Luca F, Brigante G, Perilli M, Amicosante G, et al. CTX-M-Type extended-spectrum beta-lactamases in Italy: molecular epidemiology of an emerging countrywide problem. Antimicrob Agents Chemother. 2006;50:2700-6.
(14.) Oteo J, Navarro C, Cercenado E, Delgado-Iribarren A, Wilhelmi I, Orden B, et al. Spread of Escherichia coli strains with high-level cefotaxime and ceftazidime resistance between the community, long-term care facilities, and hospital institutions. J Clin Microbiol. 2006;44:2359-66.
(15.) Pai H, Kim MR, Seo MR, Choi TY, Oh SH. A nosocomial nosocomial /noso·co·mi·al/ (nos?o-ko´me-il) pertaining to or originating in a hospital.
1. Of or relating to a hospital.
2. outbreak of Escherichia coli producing CTX-M-15 and OXA-30 beta-lactamase. Infect Control Hosp Epidemiol. 2006;27:312-4.
(16.) Pitout JD, Laupland KB, Church DL, Menard ML, Johnson JR. Virulence factors of Escherichia coli isolates that produce CTX-M-type extended-spectrum beta-lactamases. Antimicrob Agents Chemother. 2005;49:4667-70.
(17.) Lavollay M, Mamlouk K, Frank T, Akpabie A, Burghoffer B, Ben Redjeb RS, et al. Clonal dissemination of a CTX-M-15 beta-lactamase-producing Escherichia coli strain in the Paris area, Tunis, and Bangui. Antimicrob Agents Chemother. 2006;50:2433-8.
(18.) Naseer U, Natas OB, Haldorsen BC, Bue B, Grundt H, Walsh TR, et al. Nosocomial outbreak of CTX-M-15-producing E. coli in Norway. APMIS APMIS Acta Pathologica, Microbiologica et Immunologica Scandinavica
APMIS Automated Project Management Information System
APMIS Automated Project Management System . 2007;115:120-6.
(19.) Noyais A, Canton R, Moreira R, Peixe L, Baquero F, Coque TM. Emergence and dissemination of Enterobacteriaceae isolates producing CTX-M-1-like enzymes in Spain are associated with IncFII (CTX-M-15) and broad-host-range (CTX-M-1, -3, and -32) plasmids. Antimicrob Agents Chemother. 2007;51:796-9.
(20.) Sonnevend A, Al Dhaheri K, Mag T, Herpay M, Kolodziejek J, Nowotny N, et al. CTX-M-15--producing multidrug-resistant enteroaggregative Escherichia coli in the United Arab Emirates United Arab Emirates, federation of sheikhdoms (2005 est. pop. 2,563,000), c.30,000 sq mi (77,700 sq km), SE Arabia, on the Persian Gulf and the Gulf of Oman. . Clin Microbiol Infect. 2006; 12:582-5.
(21.) Lartigue MF, Fortineau N, Nordmann E Spread of novel expanded-spectrum beta-lactamases in Enterobacteriaceae in a university hospital in the Paris area, France. Clin Microbiol Infect. 2005;11: 588-91.
(22.) Lartigue MF, Zinsius C, Wenger A, Bille J, Poirel L, Nordmann P. Extended-spectrum beta-lactamases of the CTX-M type now in Switzerland. Antimicrob Agents Chemother. 2007;51:2855-60.
(23.) Brasme L, Nordmann P, Fidel F, Lartigue MF, Bajolet O, Poirel L, et al. Incidence of class A extended-spectrum (beta)-lactamases in Champagne-Ardenne (France): a 1-year prospective study. J Antimicrob Chemother. 2007;60:956-64.
(24.) Clinical and Laboratory Standards Institute. Performance standards for antimicrobial disk susceptibility tests. Approved standard. 9th ed. Document M2-A9. Wayne (PA): The Institute; 2006.
(25.) Novais A, Canton R, Valverde A, Machado E, Galan JC, Peixe L, et al. Dissemination and persistence of [bla.sub.CTX-M-9] are linked to class integrons containing CRI CRI
constant-rate infusion. associated with defective transposon transposon /trans·po·son/ (trans-po´zon) a small mobile genetic (DNA) element that moves around the genome or to other genomes within the same cell, usually by copying itself to a second site but sometimes by splicing itself out of its derivatives from Tn402 located in early antibiotic resistance plasmids of IncHI2, IncP1-alpha, and IncFI groups. Antimicrob Agents Chemother. 2006;50:2741-50.
(26.) Clermont O, Bonacorsi S, Bingen E. Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol. 2000;66:4555-8.
(27.) Johnson JR, Murray AC, Kuskowski MA, Schubert S, Prere MF, Picard B, et al.; Trans-Global Initiative for Antimicrobial Resistance Initiative (TIARA) Investigators. Distribution and characteristics of Escherichia coli clonal group A. Emerg Infect Dis. 2005; 11:141-5.
(28.) Tartof SY, Solberg OD, Manges AR, Riley LW. Analysis of a uropathogenic Escherichia coli clonal group by multilocus sequence typing. J Clin Microbiol. 2005;43:5860-4.
(29.) Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. Identification of plasmids by PCR-based replicon typing. J Microbiol Methods. 2005;63:219-28.
(30.) Robicsek A, Strahilevitz J, Jacoby GA, Macielag M, Abbanat D, Park CH, et al. Fluoroquinolone-modifying enzyme: a new adaptation of a common aminoglyeoside acetyltransferase. Nat Med. 2006;12:83-8.
(31.) Branger C, Zamfir O, Geoffroy S, Laurans G, Arlet G, Thien HV, et al. Genetic background of Escherichia eoli and extended-spectrum beta-lactamase type. Emerg Infect Dis. 2005; 11:54-61.
(32.) Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DE, et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995;33:2233-9.
(33.) Kassis-Chikhani N, Vimont S, Asselat K, Tivalle C, Minassian B, Sengelin C, et al. CTX-M beta-lactamase--producing Escherichla coli in long-term care facilities in France. Emerg Infect Dis. 2004; 10:1697-8.
(34.) Levin BR. Conditions for evolution of multiple antibiotic resistant plasmids: a theoretical and experimental excursion. In: Baumberg S, Young PW, Wellington EM, Saunders JR, editors. Population genetics of bacteria. Cambridge: Cambridge University Press Cambridge University Press (known colloquially as CUP) is a publisher given a Royal Charter by Henry VIII in 1534, and one of the two privileged presses (the other being Oxford University Press). ; 1995. p. 175-92.
(35.) Osborn AM, da Silva Tatley FM, Steyn LM, Pickup RW, Saunders JR. Mosaic plasmids and mosaic replicons: evolutionary lessons from the analysis of genetic diversity in IncFII-related replicons. Microbiology. 2000; 146:2267-75.
(36.) Szczepanowski R, Braun S, Riedel V, Schneiker S, Krahn I, Puhler A, et al. The 120 592 bp IncF plasmid pRSB107 isolated from a sewage-treatment plant encodes nine different antibiotic-resistance determinants, two iron-acquisition systems and other putative virulence-associated functions. Microbiology. 2005;151:1095-111.
(37.) Bergstrom CT, Lipsitch M, Levin BR. Natural selection, infectious transfer and the existence conditions for bacterial plasmids. Genetics. 2000;155:1505-19.
(38.) Dionisio F, Conceicao IC, Marques Marques may refer to:
(39.) Poirel L, Lartigue MF, Decousser JW, Nordmann P. ISEcp1B-mediated transposition of [bla.sub.CTX-M] in Escherichia coli. Antimicrob Agents Chemother. 2005;49:447-50.
(40.) Yagi ya·gi
n. pl. ya·gis
A directional radio and television antenna consisting of a horizontal conductor with several insulated dipoles parallel to and in the plane of the conductor. T, Kurokawa H, Senda K, Ichiyama S, Ito H, Ohsuka S, et al. Nosocomial spread of cephem-resistant Escherichia coli strains carrying multiple Toho-l-like beta-lactamase genes. Antimicrob Agents Chemother. 1997;41:2606-11.
Address for correspondence: Teresa M. Coque, Servicio de Microbiologia, Hospital Universitario Ramon y Cajal, Carretera de Colmenar, Km 9, Madrid 28034, Spain; email: firstname.lastname@example.org
Teresa M. Coque, * ([dagger])([double dagger]) Angela Novais, * ([dagger])([double dagger]) Alessandra Carattoli,([section]) Laurent Poirel, ([paragraph]) Johann Pitout, (#) ** Luisa Peixe, ([dagger])([dagger]) Fernando Baquero, * ([dagger])([double dagger]) Rafael Canton, * ([dagger])([double dagger]) and Patrice Nordmann ([paragraph])
* Hospital Universitario Ramon y Cajal, Madrid, Spain; ([dagger]) Unidad de Resistencia a Antibioticos y Virulencia Bacteriana Asociada al Consejo Superior de Investigaciones Cientificas, Madrid, Spain; ([double dagger]) El Consorcio de Investigacion Biomedica en Red de Epidemiologia y Salud Publica, Madrid, Spain; ([section]) Istituto Superiore di Sanita, Rome, Italy; ([paragraph]) Hospital Bicetre, Paris, France; (#) Calgary Laboratory Services, Calgary, Alberta, Canada; ** University of Calgary, Calgary, Alberta, Canada; and ([dagger])([dagger]) Universidade do Porto, Porto, Portugal