scholarly journals Complex Genetic Structures with Repeated Elements, a sul-Type Class 1 Integron, and the blaVEB Extended-Spectrum β-Lactamase Gene

2006 ◽  
Vol 50 (5) ◽  
pp. 1745-1752 ◽  
Author(s):  
Thierry Naas ◽  
Daniel Aubert ◽  
Thierry Lambert ◽  
Patrice Nordmann
Keyword(s):  
Direct Repeat ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Gene Cassettes ◽  
Conserved Sequence ◽  
Extended Spectrum ◽  
Class 1 ◽  
Genetic Structures ◽  
Lactamase Gene ◽  
First Time

ABSTRACT Two clinical isolates of Pseudomonas aeruginosa, TL-1 and TL-2, were isolated from a patient transferred from Bangladesh and hospitalized for osteomyelitis in Paris, France. P. aeruginosa TL-1 expressed the extended-spectrum β-lactamase VEB-1a and was susceptible only to imipenem and colistin, while P. aeruginosa TL-2 expressed only the naturally occurring bla AmpC gene at a basal level and exhibited a wild-type β-lactam resistance phenotype. In TL-1, the typical 5′-end conserved sequence (5′-CS) region of class 1 integrons usually present upstream of the bla VEB-1a gene was replaced by a truncated 3′-CS and a 135-bp repeated element (Re). Downstream of the bla VEB-1a gene, an insertion sequence, ISPa31 disrupted by ISPa30, and an orf513 sequence, belonging to a common region (conserved region 1 [CR1]) immediately upstream of the aphA-6 gene, were present. Further downstream, a second truncated 3′-CS region in direct repeat belonged to In51, an integron containing two gene cassettes (aadA6 and the OrfD cassette). Thus, the overall structure corresponded to a sul-type class 1 integron termed In121. Genetic analyses revealed that both isolates were clonally related and differed by a ca. 100-kb fragment that contained In121. Both isolates contained another integron, In122, that carried three gene cassettes: aadB, dfrA1, and the OrfX cassette. This work identifies for the first time the spread of Re-associated bla VEB genes located on a sul-type integron. It also reports for the first time a CR1 element in P. aeruginosa that is associated with an aminoglycoside resistance aphA-6 gene that is expressed from a composite promoter.

scholarly journals Functional and Structural Characterization of the Genetic Environment of an Extended-Spectrum β-Lactamase blaVEB Gene from a Pseudomonas aeruginosa Isolate Obtained in India

2004 ◽  
Vol 48 (9) ◽  
pp. 3284-3290 ◽  
Author(s):  
Daniel Aubert ◽  
Delphine Girlich ◽  
Thierry Naas ◽  
Shanta Nagarajan ◽  
Patrice Nordmann
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Dna Sequences ◽  
Direct Repeat ◽  
Clinical Strain ◽  
Gene Encoding ◽  
Class 1 Integron ◽  
Genetic Environment ◽  
Conserved Sequence ◽  
Extended Spectrum ◽  
Class 1

ABSTRACT A Pseudomonas aeruginosa clinical strain isolated from a patient hospitalized in a New Delhi, India, hospital was resistant to expanded-spectrum cephalosporins, imipenem, and aztreonam. A bla VEB-1-like gene named bla VEB-1a, which codes for the extended-spectrum β-lactamase VEB-1a, was identified. The genetic environment of bla VEB-1a was peculiar: (i) no 5′ conserved sequence (5′-CS) region was present upstream of the β-lactamase gene, whereas bla VEB-1-like genes are usually associated with class 1 integrons; (ii) bla VEB-1a was inserted between two truncated 3′-CS regions in a direct repeat; and (iii) four 135-bp repeated DNA sequences (repeated elements) were located on each side of the bla VEB-1a gene. Expression of the bla VEB-1a gene was driven by a strong promoter located in one of these repeated sequences. In addition, cloning of the β-lactamase content of this P. aeruginosa isolate followed by expression in Escherichia coli identified the naturally occurring AmpC β-lactamase and a gene encoding an OXA-2-like β-lactamase located in a class 1 integron, In78, in which an insertion sequence, ISpa7, was inserted within its 5′-CS region.

Antimicrobial Resistance, Extended-Spectrum β-Lactamase Productivity, and Class 1 Integrons in Escherichia coli from Healthy Swine

2015 ◽  
Vol 78 (8) ◽  
pp. 1442-1450 ◽  
Author(s):  
KANJANA CHANGKAEW ◽  
APIRADEE INTARAPUK ◽  
FUANGFA UTRARACHKIJ ◽  
CHIE NAKAJIMA ◽  
ORASA SUTHIENKUL ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Intestinal Flora ◽  
Class 1 Integron ◽  
Potential Health ◽  
Gene Cassettes ◽  
E Coli ◽  
Class 1 Integrons ◽  
Extended Spectrum ◽  
Class 1

Administration of antimicrobials to food-producing animals increases the risk of higher antimicrobial resistance in the normal intestinal flora of these animals. The present cross-sectional study was conducted to investigate antimicrobial susceptibility and extended-spectrum β-lactamase (ESBL)–producing strains and to characterize class 1 integrons in Escherichia coli in healthy swine in Thailand. All 122 of the tested isolates had drug-resistant phenotypes. High resistance was found to ampicillin (98.4% of isolates), chloramphenicol (95.9%), gentamicin (78.7%), streptomycin (77.9%), tetracycline (74.6%), and cefotaxime (72.1%). Fifty-four (44.3%) of the E. coli isolates were confirmed as ESBL-producing strains. Among them, blaCTX-M (45 isolates) and blaTEM (41 isolates) were detected. Of the blaCTX-M-positive E. coli isolates, 37 carried the blaCTX-M-1 cluster, 12 carried the blaCTX-M-9 cluster, and 5 carried both clusters. Sequence analysis revealed blaTEM-1, blaTEM-135, and blaTEM-175 in 38, 2, and 1 isolate, respectively. Eighty-seven (71%) of the 122isolates carried class 1 integrons, and eight distinct drug-resistance gene cassettes with seven different integron profiles were identified in 43 of these isolates. Gene cassettes were associated with resistance to aminoglycosides (aadA1, aadA2, aadA22, or aadA23), trimethoprim (dfrA5, dfrA12, or dfrA17), and lincosamide (linF). Genes encoding β-lactamases were not found in class 1 integrons. This study is the first to report ESBL-producing E. coli with a class 1 integron carrying the linF gene cassette in swine in Thailand. Our findings confirm that swine can be a reservoir of ESBL-producing E. coli harboring class 1 integrons, which may become a potential health risk if these integrons are transmitted to humans. Intensive analyses of animal, human, and environmental isolates are needed to control the spread of ESBL-producing E. coli strains.

scholarly journals The aadB Gene Cassette Is Associated with blaSHV Genes in Klebsiella Species Producing Extended-Spectrum β-Lactamases

2005 ◽  
Vol 49 (2) ◽  
pp. 794-797 ◽  
Author(s):  
Louisa A. Jones ◽  
Christopher J. McIver ◽  
Mi-Jurng Kim ◽  
William D. Rawlinson ◽  
Peter A. White
Keyword(s):  
Gene Cassette ◽  
Amplicon Sequencing ◽  
Klebsiella Species ◽  
Class 1 Integron ◽  
Beta Lactamases ◽  
Gene Cassettes ◽  
Extended Spectrum ◽  
Class 1 ◽  
Extended Spectrum Beta Lactamases

ABSTRACT Integrons were detected in 37 (72.5%) of 51 Klebsiella spp. producing extended-spectrum beta-lactamases by PCR with primers that targeted integrase genes and cassette regions. PCR and amplicon sequencing of the cassette regions revealed aadB and aadA2 gene cassettes that confer resistance to a range of aminoglycosides. aadB was associated with a class 1 integron on a 28-kb plasmid, pES1, that also contained bla SHV-12 and IS26.

scholarly journals Class 1 Integron-Associated Tobramycin-Gentamicin Resistance in Campylobacter jejuni Isolated from the Broiler Chicken House Environment

2002 ◽  
Vol 46 (11) ◽  
pp. 3660-3664 ◽  
Author(s):  
Margie D. Lee ◽  
Susan Sanchez ◽  
Martha Zimmer ◽  
Umelaalim Idris ◽  
Mark E. Berrang ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Campylobacter Jejuni ◽  
Broiler Chicken ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Integrase Gene ◽  
Conserved Sequence ◽  
Chicken House ◽  
Class 1 ◽  
Gentamicin Resistance

ABSTRACT Using PCR, we screened 105 isolates of poultry-associated Campylobacter jejuni for the presence of class 1 integrons. Of those isolates, 21% (22 of 105) possessed the integrase gene, but only 5 isolates produced an amplicon in a 5′-3′ conserved sequence PCR directed toward amplification of the resistance cassettes. DNA sequencing demonstrated that all five isolates possessed the aminoglycoside resistance gene, aacA4.

scholarly journals New Determinants of Aminoglycoside Resistance and Their Association with the Class 1 Integron Gene Cassettes in Trueperella pyogenes

2020 ◽  
Vol 21 (12) ◽  
pp. 4230
Author(s):  
Ewelina Kwiecień ◽  
Ilona Stefańska ◽  
Dorota Chrobak-Chmiel ◽  
Agnieszka Sałamaszyńska-Guz ◽  
Magdalena Rzewuska
Keyword(s):  
Resistance Genes ◽  
Resistance Mechanisms ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Gene Cassettes ◽  
Genetic Elements ◽  
Trueperella Pyogenes ◽  
Broth Microdilution Method ◽  
Class 1 ◽  
Aminoglycoside Resistance Genes

Trueperella pyogenes is an important opportunistic animal pathogen. Different antimicrobials, including aminoglycosides, are used to treat T. pyogenes infections. The aim of the present study was to evaluate aminoglycoside susceptibility and to detect aminoglycoside resistance determinants in 86 T. pyogenes isolates of different origin. Minimum inhibitory concentration of gentamicin, streptomycin, and kanamycin was determined using a standard broth microdilution method. Genetic elements associated with aminoglycoside resistance were investigated by PCR and DNA sequencing. All studied isolates were susceptible to gentamicin, but 32.6% and 11.6% of them were classified as resistant to streptomycin and kanamycin, respectively. A total of 30 (34.9%) isolates contained class 1 integrons. Class 1 integron gene cassettes carrying aminoglycoside resistance genes, aadA11 and aadA9, were found in seven and two isolates, respectively. Additionally, the aadA9 gene found in six isolates was not associated with mobile genetic elements. Moreover, other, not carried by gene cassettes, aminoglycoside resistance genes, strA-strB and aph(3’)-IIIa, were also detected. Most importantly, this is the first description of all reported genes in T. pyogenes. Nevertheless, the relevance of the resistance phenotype to genotype was not perfectly matched in 14 isolates. Therefore, further investigations are needed to fully explain aminoglycoside resistance mechanisms in T. pyogenes.

scholarly journals Chromosome-Encoded AmpC and CTX-M Extended-Spectrum β-Lactamases in Clinical Isolates ofProteus mirabilisfrom Korea

2011 ◽  
Vol 55 (4) ◽  
pp. 1414-1419 ◽  
Author(s):  
Wonkeun Song ◽  
Juwon Kim ◽  
Il Kwon Bae ◽  
Seok Hoon Jeong ◽  
Young Hee Seo ◽  
...  
Keyword(s):  
Chromosomal Location ◽  
Clinical Isolates ◽  
Class 1 Integron ◽  
Extended Spectrum ◽  
Hospital Environments ◽  
Class 1 ◽  
Genes Encoding ◽  
Unusual Phenomenon ◽  
M 12 ◽  
Lactamase Gene

ABSTRACTAmong 222Proteus mirabilisclinical isolates collected from 17 hospitals in Korea in 2008, 28 (12.6%) and 8 (3.6%) isolates exhibited extended-spectrum β-lactamase (ESBL) and AmpC phenotypes, respectively. The most common type of ESBL gene identified by PCR and sequencing experiments wasblaCTX-M-14a(n= 12). TheblaCTX-M-90(n= 4),blaCTX-M-15(n= 3),blaCTX-M-12(n= 3),blaCTX-M-2(n= 2),blaCTX-M-14b(n= 1),blaTEM-52(n= 5), andblaSHV-12(n= 1) genes were also detected. Eight isolates carried an AmpC β-lactamase gene, such asblaCMY-2(n= 6) orblaDHA-1(n= 2). Allblagenes encoding CTX-M-1- and CTX-M-9-type enzymes and allblaCMY-2genes were preceded by ISEcp1-like elements. TheblaCTX-M-2gene found in two isolates was located on a complex class 1 integron. TheblaDHA-1gene was preceded by a transcriptional regulator gene and was followed by phage shock protein genes. TheblaCTX-Mgenes were located on the chromosome in 21 isolates. A plasmid location for theblaCTX-Mgene was found in only four isolates: theblaCTX-M-14agene was located on ∼150-kbp IncA/C plasmids in three isolates and on a ∼50-kbp IncN plasmid in one isolate. TheblaTEM-52gene was located on ∼50-kbp IncN plasmids in all five isolates. The AmpC β-lactamase genes were located on the chromosome in seven of eight isolates; one isolate carried theblaCMY-2gene on a ∼150-kbp IncA/C plasmid. Our results show that a chromosomal location of CTX-M ESBL and AmpC β-lactamase genes inP. mirabilisis no longer an unusual phenomenon in hospital environments.

scholarly journals Molecular Characterization of a Novel Class 1 Integron Containing blaGES-1 and a Fused Product of aac(3)-Ib/aac(6")-Ib" Gene Cassettes in Pseudomonas aeruginosa

2002 ◽  
Vol 46 (3) ◽  
pp. 638-645 ◽  
Author(s):  
Véronique Dubois ◽  
Laurent Poirel ◽  
Caroline Marie ◽  
Corinne Arpin ◽  
Patrice Nordmann ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Diffusion Method ◽  
Fusion Product ◽  
Resistance Pattern ◽  
Clinical Strain ◽  
Homologous Sequence ◽  
Class 1 Integron ◽  
Gene Cassettes ◽  
Extended Spectrum ◽  
Class 1

ABSTRACT As seen by the disk diffusion method, the clinical strain of Pseudomonas aeruginosa Pa695, resistant to all extended-spectrum cephalosporins and aminoglycosides, exhibited an unusual synergistic effect between ceftazidime and imipenem. This isolate produced an extended-spectrum β-lactamase (ESBL) with a pI of 5.8 that appeared to be chromosomally encoded. Cloning experiments revealed that this ESBL was encoded by bla GES-1, previously described in an integron from Klebsiella pneumoniae. In P. aeruginosa Pa695, a higher level of resistance to ceftazidime than to ticarcillin was observed, and no synergy between the β-lactamase inhibitors and extended-spectrum cephalosporins was detected, in contrast to the resistance pattern observed in K. pneumoniae. Further sequence analysis demonstrated that the bla GES-1 gene cassette was located in a class 1 integron, which contained another sequence corresponding to the fused aac(3)-Ib and aac(6")-Ib" gene cassettes. The fusion product was functional, as was the product of each gene cloned separately: AAC(3)-I, despite the deletion of the four last amino acids, and AAC(6"), which carried three amino acid changes compared with the most homologous sequence. The AAC(3)-I protein conferred an expected gentamicin and fortimicin resistance, and the AAC(6"), despite the Leu-119→Ser substitution, yielded resistance to kanamycin, tobramycin, and dibekacin, but slightly affected netilmicin and amikacin, and had no apparent effect on gentamicin. The fusion product conveyed a large profile of resistance, combining the AAC(6") activity with a higher level of gentamicin resistance without accompanying fortimicin resistance.

scholarly journals Characterization of Class 1 Integrons from Pseudomonas aeruginosa That Contain the blaVIM-2Carbapenem-Hydrolyzing β-Lactamase Gene and of Two Novel Aminoglycoside Resistance Gene Cassettes

2001 ◽  
Vol 45 (2) ◽  
pp. 546-552 ◽  
Author(s):  
Laurent Poirel ◽  
Thierry Lambert ◽  
Salih Türkoglü ◽  
Esthel Ronco ◽  
Jean-Louis Gaillard ◽  
...  
Keyword(s):  
Amino Acids ◽  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Resistance Gene ◽  
Gene Cassette ◽  
Amino Acid Sequences ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Gene Cassettes ◽  
Class 1

ABSTRACT Two clonally unrelated Pseudomonas aeruginosa clinical strains, RON-1 and RON-2, were isolated in 1997 and 1998 from patients hospitalized in a suburb of Paris, France. Both isolates expressed the class B carbapenem-hydrolyzing β-lactamase VIM-2 previously identified in Marseilles in the French Riviera. In both isolates, thebla VIM-2 cassette was part of a class 1 integron that also encoded aminoglycoside-modifying enzymes. In one case, two novel aminoglycoside resistance gene cassettes,aacA29a and aacA29b, were located at the 5′ and 3′ end of the bla VIM-2 gene cassette, respectively. The aacA29a and aacA29b gene cassettes were fused upstream with a 101-bp part of the 5′ end of theqacE cassette. The deduced amino acid sequence AAC(6′)-29a protein shared 96% identity with AAC(6′)-29b but only 34% identity with the aacA7-encoded AAC(6′)-I1, the closest relative of the AAC(6′)-I family enzymes. These aminoglycoside acetyltransferases had amino acid sequences much shorter (131 amino acids) than the other AAC(6′)-I enzymes (144 to 153 amino acids). They conferred resistance to amikacin, isepamicin, kanamycin, and tobramycin but not to gentamicin, netilmicin, and sisomicin.

scholarly journals bla VIM-2-Harboring Integrons Isolated in India, Russia, and the United States Arise from an Ancestral Class 1 Integron Predating the Formation of the 3′ Conserved Sequence

2007 ◽  
Vol 51 (7) ◽  
pp. 2636-2638 ◽  
Author(s):  
Mark A. Toleman ◽  
Hemalatha Vinodh ◽  
Uma Sekar ◽  
Vijaylakshmi Kamat ◽  
Timothy R. Walsh
Keyword(s):  
United States ◽  
Pseudomonas Aeruginosa ◽  
The United States ◽  
Class 1 Integron ◽  
Conserved Sequence ◽  
Class 1 ◽  
Lactamase Gene

ABSTRACT The metallo-β-lactamase gene bla VIM-2 was identified in a strain of Pseudomonas aeruginosa isolated in India. The integron encoding bla VIM-2 was virtually identical to those recently found in the United States and Russia. These unusual structures are likely to have arisen from an ancestral integron predating the formation of the 3′ conserved sequence.

scholarly journals Biochemical Sequence Analyses of GES-1, a Novel Class A Extended-Spectrum β-Lactamase, and the Class 1 Integron In52 from Klebsiella pneumoniae

2000 ◽  
Vol 44 (3) ◽  
pp. 622-632 ◽  
Author(s):  
Laurent Poirel ◽  
Isabelle Le Thomas ◽  
Thierry Naas ◽  
Amal Karim ◽  
Patrice Nordmann
Keyword(s):  
Amino Acid ◽  
Klebsiella Pneumoniae ◽  
Amino Acid Identity ◽  
Class 1 Integron ◽  
Chloramphenicol Resistance ◽  
Acid Identity ◽  
Gene Cassettes ◽  
Class A ◽  
Extended Spectrum ◽  
Class 1

ABSTRACT Klebsiella pneumoniae ORI-1 was isolated in 1998 in France from a rectal swab of a 1-month-old girl who was previously hospitalized in Cayenne Hospital, Cayenne, French Guiana. This strain harbored a ca. 140-kb nontransferable plasmid, pTK1, that conferred an extended-spectrum cephalosporin resistance profile antagonized by the addition of clavulanic acid, tazobactam, or imipenem. The gene for GES-1 (Guiana extended-spectrum β-lactamase) was cloned, and its protein was expressed in Escherichia coli DH10B, where this pI-5.8 β-lactamase of a ca. 31-kDa molecular mass conferred resistance to oxyimino cephalosporins (mostly to ceftazidime). GES-1 is weakly related to the other plasmid-located Ambler class A extended-spectrum β-lactamases (ESBLs). The highest percentage of amino acid identity was obtained with the carbenicillinase GN79 from Proteus mirabilis; with YENT, a chromosome-borne penicillinase fromYersinia enterocolitica; and with L-2, a chromosome-borne class A cephalosporinase from Stenotrophomonas maltophilia(36% amino acid identity each). However, a dendrogram analysis showed that GES-1 clustered within a class A ESBL subgroup together with ESBLs VEB-1 and PER-1. Sequencing of a 7,098-bp DNA fragment from plasmid pTK1 revealed that the GES-1 gene was located on a novel class 1 integron named In52 that was characterized by (i) a 5′ conserved segment containing an intI1 gene possessing two putative promoters, P1 and P2, for coordinated expression of the downstream antibiotic resistance genes and an attI1 recombination site; (ii) five antibiotic gene cassettes, bla GES-1,aac(6′)Ib′ (gentamicin resistance and amikacin susceptibility), dfrXVb (trimethoprim resistance), a novel chloramphenicol resistance gene (cmlA4), andaadA2 (streptomycin-spectinomycin resistance); and (iii) a 3′ conserved segment consisting of qacEΔ1 andsulI. The bla GES-1 andaadA2 gene cassettes were peculiar, since they lacked a typical 59-base element. This work identified the second class A ESBL gene of a non-TEM, non-SHV series which was located in the plasmid and integron, thus providing it additional means for its spread and its expression.

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