scholarly journals Characterization of the New Metallo-β-Lactamase VIM-13 and Its Integron-Borne Gene from a Pseudomonas aeruginosa Clinical Isolate in Spain

2008 ◽  
Vol 52 (10) ◽  
pp. 3589-3596 ◽  
Author(s):  
Carlos Juan ◽  
Alejandro Beceiro ◽  
Olivia Gutiérrez ◽  
Sebastián Albertí ◽  
Margalida Garau ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Pcr Amplification ◽  
Class 1 Integron ◽  
Negative Results ◽  
New Variant ◽  
Class B ◽  
Class 1 ◽  
Encoding Gene ◽  
Good Agreement

ABSTRACT During a survey conducted to evaluate the incidence of class B carbapenemase (metallo-β-lactamase [MBL])-producing Pseudomonas aeruginosa strains from hospitals in Majorca, Spain, five clinical isolates showed a positive Etest MBL screening test result. In one of them, strain PA-SL2, the presence of a new bla VIM derivative (bla VIM-13) was detected by PCR amplification with bla VIM-1-specific primers followed by sequencing. The bla VIM-13-producing isolate showed resistance to all β-lactams (except aztreonam), gentamicin, tobramycin, and ciprofloxacin. VIM-13 exhibited 93% and 88% amino acid sequence identities with VIM-1 and VIM-2, respectively. bla VIM-13 was cloned in parallel with bla VIM-1, and the resistance profile conferred was analyzed both in Escherichia coli and in P. aeruginosa backgrounds. Compared to VIM-1, VIM-13 conferred slightly higher levels of resistance to piperacillin and lower levels of resistance to ceftazidime and cefepime. VIM-13 and VIM-1 were purified in parallel as well, and their kinetic parameters were compared. The k cat/K m ratios for the antibiotics mentioned above were in good agreement with the MIC data. Furthermore, EDTA inhibited the activity of VIM-13 approximately 25 times less than it inhibited the activity of VIM-1. VIM-13 was harbored in a class 1 integron, along with a new variant (Ala108Thr) of the aminoglycoside-modifying enzyme encoding gene aacA4, which confers resistance to gentamicin and tobramycin. Finally, the VIM-13 integron was apparently located in the chromosome, since transformation and conjugation experiments consistently yielded negative results and the bla VIM-13 probe hybridized only with the genomic DNA.

scholarly journals Molecular Epidemiology and Mechanisms of Carbapenem Resistance in Pseudomonas aeruginosa Isolates from Spanish Hospitals

2007 ◽  
Vol 51 (12) ◽  
pp. 4329-4335 ◽  
Author(s):  
O. Gutiérrez ◽  
C. Juan ◽  
E. Cercenado ◽  
F. Navarro ◽  
E. Bouza ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Carbapenem Resistance ◽  
Clonal Diversity ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Carbapenem Resistant ◽  
Class B ◽  
Class 1 ◽  
Encoding Gene ◽  
Low Prevalence

ABSTRACT All (236) Pseudomonas aeruginosa isolates resistant to imipenem and/or meropenem collected during a multicenter (127-hospital) study in Spain were analyzed. Carbapenem-resistant isolates were found to be more frequently resistant to all β-lactams and non-β-lactam antibiotics than carbapenem-susceptible isolates (P < 0.001), and up to 46% of the carbapenem-resistant isolates met the criteria used to define multidrug resistance (MDR). Pulsed-field gel electrophoresis revealed remarkable clonal diversity (165 different clones were identified), and with few exceptions, the levels of intra- and interhospital dissemination of clones were found to be low. Carbapenem resistance was driven mainly by the mutational inactivation of OprD, accompanied or not by the hyperexpression of AmpC or MexAB-OprM. Class B carbapenemases (metallo-β-lactamases [MBLs]) were detected in a single isolate, although interestingly, this isolate belonged to one of the few epidemic clones documented. The MBL-encoding gene (bla VIM-2), along with the aminoglycoside resistance determinants, was transferred to strain PAO1 by electroporation, demonstrating its plasmid location. The class 1 integron harboring bla VIM-2 was characterized as well, and two interesting features were revealed: intI1 was found to be disrupted by a 1.1-kb insertion sequence, and a previously undescribed aminoglycoside acetyltransferase-encoding gene [designated aac(6′)-32] preceded bla VIM-2. AAC(6′)-32 showed 80% identity to AAC(6′)-Ib′ and the recently described AAC(6′)-31, and when aac(6′)-32 was cloned into Escherichia coli, it conferred resistance to tobramycin and reduced susceptibility to gentamicin and amikacin. Despite the currently low prevalence of epidemic clones with MDR, active surveillance is needed to detect and prevent the dissemination of these clones, particularly those producing integron- and plasmid-encoded MBLs, given their additional capacity for the intra- and interspecies spread of MDR.

Characterization of VIM-4 Producing Clinical Pseudomonas aeruginosa Isolates from Western Algeria: Sequence Type and Class 1 Integron Description

2020 ◽  
Vol 26 (12) ◽  
pp. 1437-1441
Author(s):  
Fatma Zohra Zaidi ◽  
Radia Dali-Yahia ◽  
Karima Zenati ◽  
Leila Yazi ◽  
Manon Lounes ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Sequence Type ◽  
Class 1 Integron ◽  
Class 1 ◽  
Western Algeria

scholarly journals Detection and Characterization of VIM-31, a New Variant of VIM-2 with Tyr224His and His252Arg Mutations, in a Clinical Isolate of Enterobacter cloacae

2012 ◽  
Vol 56 (6) ◽  
pp. 3283-3287 ◽  
Author(s):  
Pierre Bogaerts ◽  
Carine Bebrone ◽  
Te-Din Huang ◽  
Warda Bouchahrouf ◽  
Yves DeGheldre ◽  
...  
Keyword(s):  
Catalytic Efficiency ◽  
Enterobacter Cloacae ◽  
Colonic Adenocarcinoma ◽  
Beta Lactam ◽  
Class 1 Integron ◽  
Content Type ◽  
New Variant ◽  
Class 1 ◽  
Blood Specimens

ABSTRACTWe report the first description of the metallo-β-lactamase VIM-31, a new variant of VIM-2 with Tyr224His and His252Arg mutations, inEnterobacter cloacae11236, which was isolated from blood specimens of a patient with colonic adenocarcinoma in Belgium.blaVIM-31was found on a class 1 integron located on a self-transferable but not typeable 42-kb plasmid. Compared to values published elsewhere for VIM-2, the purified VIM-31 enzyme showed weaker catalytic efficiency against all the tested beta-lactam agents (except for ertapenem), resulting from lowerkcat(except for ertapenem) and higherKmvalues for VIM-31.

scholarly journals Genetic characterization of Shigella spp. isolated from diarrhoeal and asymptomatic children

2014 ◽  
Vol 63 (7) ◽  
pp. 903-910 ◽  
Author(s):  
Santanu Ghosh ◽  
Gururaja P. Pazhani ◽  
Swapan Kumar Niyogi ◽  
James P. Nataro ◽  
Thandavarayan Ramamurthy
Keyword(s):  
Virulence Genes ◽  
Multidrug Resistant ◽  
Class 1 Integron ◽  
Genetic Characteristics ◽  
Asymptomatic Children ◽  
Class 1 ◽  
Shigella Spp ◽  
Encoding Gene ◽  
Encoding Genes

Phenotypic and genetic characteristics of Shigella spp. isolated from diarrhoeal and asymptomatic children aged up to 5 years were analysed in this study. In total, 91 and 17 isolates were identified from diarrhoeal (case) and asymptomatic (control) children, respectively. All the isolates were tested for antimicrobial resistance, the presence of integrons, plasmid-mediated quinolone resistance (PMQR), virulence-associated genes and Shigella pathogenicity island (SH-PAI). The majority of the Shigella spp. from cases (68.1 %) and controls (82.3 %) were found to be resistant to fluoroquinolones. Integron carriage was detected more in cases (76.9 %) than in controls (35.5 %). Atypical class 1 integron was detected exclusively in Shigella flexneri from cases but not from the controls. PMQR genes such as aac(6′)-Ib-cr and qnrS1 were detected in 82.4  and 14.3 % of the isolates from cases and in 53 and 17.6 % in controls, respectively. Shigella isolates from cases as well as from controls were positive for the invasive plasmid antigen H-encoding gene ipaH. The other virulence genes such as virF, sat, setA, setB, sen and ial were detected in Shigella isolates in 80.2, 49.4, 27.4, 27.4, 80.2 and 79.1 % of cases and in 64.7, 52.9, 17.6, 17.6, 64.7 and 64.7 % of controls, respectively. The entire SH-PAI was detected in S. flexneri serotype 2a from cases and controls. In an isolate from a control child, the SH-PAI was truncated. Integrons, PMQR and virulence-encoding genes were detected more frequently in cases than in controls. In diarrhoea endemic areas, asymptomatic carriers may play a crucial role in the transmission of multidrug-resistant Shigella spp. with all the putative virulence genes.

scholarly journals bla VIM-2 Cassette-Containing Novel Integrons in Metallo-β-Lactamase-Producing Pseudomonas aeruginosa and Pseudomonas putida Isolates Disseminated in a Korean Hospital

2002 ◽  
Vol 46 (4) ◽  
pp. 1053-1058 ◽  
Author(s):  
Kyungwon Lee ◽  
Jong Back Lim ◽  
Jong Hwa Yum ◽  
Dongeun Yong ◽  
Yunsop Chong ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Pseudomonas Putida ◽  
Tertiary Care ◽  
Care Hospital ◽  
Class 1 Integron ◽  
Class B ◽  
Class 1 ◽  
Horizontal Spread ◽  
Imipenem Resistance ◽  
O 11

ABSTRACT We investigated the phenotypic and genetic properties of metallo-β-lactamase-producing Pseudomonas isolates collected at a tertiary-care hospital in Korea since 1995. The prevalence of imipenem resistance among Pseudomonas aeruginosa isolates reached 16% in 1997, when 9% of the resistant organisms were found to produce VIM-2 β-lactamase, a class B enzyme previously found only in P. aeruginosa isolates from Europe. VIM-2-producing isolates of Pseudomonas putida were also detected. Resistance was transferable from both these species to P. aeruginosa PAO4089Rp by filter mating, although the resistance determinant could not be found on any detectable plasmid. Serotyping showed that many of the VIM-2-producing P. aeruginosa isolates belonged to serotypes O:11 and O:12, and pulsed-field gel electrophoresis of XbaI-digested genomic DNA revealed that many had identical profiles, whereas the P. putida isolates were diverse. Sequencing showed that the bla VIM-2 genes resided as cassettes in class 1 integrons. In contrast to previous VIM-encoding integrons, the integron sequenced from a P. aeruginosa isolate had bla VIM located downstream of a variant of aacA4. bla VIM also lay in a class 1 integron in a representative P. putida strain, but the organization of this integron was different from that sequenced from the P. aeruginosa strain. In conclusion, the metallo-β-lactamase produced by these imipenem-resistant Pseudomonas isolates was VIM-2, and the accumulation of producers reflected clonal dissemination as well as horizontal spread. Strict measures are required in order to control a further spread of resistance.

INTEGRON DIVERSITY IN BLAVIM-2-CARRYING CARBAPENEM-RESISTANT CLINICAL PSEUDOMONAS AERUGINOSA ISOLATES

2019 ◽  
Vol 64 (8) ◽  
pp. 497-502 ◽  
Author(s):  
T. A. Savinova ◽  
Yu. A. Bocharova ◽  
A. V. Lazareva ◽  
I. V. Chebotar ◽  
N. A. Mayanskiy
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Gene Cassette ◽  
Variable Regions ◽  
Class 1 Integrons ◽  
Carbapenem Resistant ◽  
Class 1 ◽  
Encoding Gene ◽  
Clonal Spread ◽  
Pathogen Populations

The growing prevalence of metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa in nosocomial pathogen populations has been attributed to their clonal spread, and/or horizontal transfer of MBL determinants in mobile genetic elements, including integrons. To characterize the genetic background of the beta-lactamase VIM-2 encoding gene in the population of carbapenem-resistant (Carba-R) P. aeruginosa clinical isolates.The detection of class 1 integrons was performed by PCR. Typing of the class 1 integrons containing the blaVIM gene cassette was performed by the PCR-restriction fragment length polymorphism (RFLP) approach followed by sequencing of variable regions of class 1 integrons. Five types of the blaVIM-2-carrying integrons were identified: ST654-isolates accounting for more than 50% of the Carba-R population harbored In56; ST235-isolates contained In559 (26% Carba-R isolates); ST111-isolates (19% Carba-R isolates) were characterized by carrying In59-like integron; two ST235-isolates harbored In59 and In249 each. Except In56, carrying the only blaVIM-2-gene cassette, all other identified integron types harbored the genes of resistance to trimethoprim and/or aminoglycosides. No new types of integrons were identified in the P. aeruginosa clinical isolates. The observed correlation of the integron type with specific STs indicates a clonal dissemination of significant resistance determinant producers - ST111, ST654 and ST235 epidemic lines. The features of the integron variable regions can be used for the epidemiological characterization of clinical P. aeruginosa isolates.

scholarly journals AAC(6′)-Iaf, a Novel Aminoglycoside 6′-N-Acetyltransferase from Multidrug-Resistant Pseudomonas aeruginosa Clinical Isolates

2009 ◽  
Vol 53 (6) ◽  
pp. 2327-2334 ◽  
Author(s):  
Tomoe Kitao ◽  
Tohru Miyoshi-Akiyama ◽  
Teruo Kirikae
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Decubitus Ulcer ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Upstream Region ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Class 1 ◽  
Layer Chromatography

ABSTRACT We report here the characterization of a novel aminoglycoside resistance gene, aac(6′)-Iaf, present in two multidrug-resistant (MDR) Pseudomonas aeruginosa clinical isolates. These isolates, IMCJ798 and IMCJ799, were independently obtained from two patients, one with a urinary tract infection and the other with a decubitus ulcer, in a hospital located in the western part of Japan. Although the antibiotic resistance profiles of IMCJ798 and IMCJ799 were similar to that of MDR P. aeruginosa IMCJ2.S1, which caused outbreaks in the eastern part of Japan, the pulsed-field gel electrophoresis patterns for these isolates were different from that for IMCJ2.S1. Both IMCJ798 and IMCJ799 were found to contain a novel chromosomal class 1 integron, In123, which included aac(6′)-Iaf as the first cassette gene. The encoded protein, AAC(6′)-Iaf, was found to consist of 183 amino acids, with 91 and 87% identity to AAC(6′)-Iq and AAC(6′)-Im, respectively. IMCJ798, IMCJ799, and Escherichia coli transformants carrying a plasmid containing the aac(6′)-Iaf gene and its upstream region were highly resistant to amikacin, dibekacin, and kanamycin but not to gentamicin. The production of AAC(6′)-Iaf in these strains was confirmed by Western blot analysis. Thin-layer chromatography indicated that AAC(6′)-Iaf is a functional acetyltransferase that specifically modifies the amino groups at the 6′ positions of aminoglycosides. Collectively, these findings indicate that AAC(6′)-Iaf contributes to aminoglycoside resistance.

scholarly journals Characterization of DIM-1, an Integron-Encoded Metallo-β-Lactamase from a Pseudomonas stutzeri Clinical Isolate in the Netherlands

2010 ◽  
Vol 54 (6) ◽  
pp. 2420-2424 ◽  
Author(s):  
Laurent Poirel ◽  
Jose-Manuel Rodríguez-Martínez ◽  
Nashwan Al Naiemi ◽  
Yvette J. Debets-Ossenkopp ◽  
Patrice Nordmann
Keyword(s):  
Gc Content ◽  
Pseudomonas Stutzeri ◽  
Amino Acid Identity ◽  
Class 1 Integron ◽  
Acid Identity ◽  
Carbapenem Resistant ◽  
Class B ◽  
Class 1 ◽  
Dutch Patient

ABSTRACT A carbapenem-resistant Pseudomonas stutzeri strain isolated from a Dutch patient was analyzed in detail. This isolate produced a metallo-β-lactamase (MBL) whose gene, with 43.5% GC content, was cloned and expressed in Escherichia coli. β-Lactamase DIM-1 (for Dutch imipenemase) was weakly related to other Ambler class B β-lactamases, sharing <52% amino acid identity with the most closely related MBL, GIM-1, and 45% identity with IMP-type MBLs. The β-Lactamase DIM-1 significantly hydrolyzed broad-spectrum cephalosporins and carbapenems and spared aztreonam. This MBL gene was embedded in a class 1 integron containing two other gene cassettes, encoding resistance to aminoglycosides and disinfectants, that was located on a 70-kb plasmid.

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