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

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.

2006 ◽  
Vol 135 (3) ◽  
pp. 502-504 ◽  
Author(s):  
T. CAETANO ◽  
S. FERREIRA ◽  
A. P. MONDEGO ◽  
A. CORREIA ◽  
S. MENDO

In99, a possible ancestor of In100, is a class 1 integron associated with carbenicillinase (blaPSE) and aminoglycoside resistance genes [aac(6′)-Ib and aadA2]. In99 was present in 8 of 81 clinical isolates of Pseudomonas aeruginosa from unrelated patients collected in different years. The strains fell into two clonal groups and exhibited resistance to β-lactams and aminoglycosides.


2012 ◽  
Vol 57 (1) ◽  
pp. 96-100 ◽  
Author(s):  
Tatsuya Tada ◽  
Tohru Miyoshi-Akiyama ◽  
Kayo Shimada ◽  
Masahiro Shimojima ◽  
Teruo Kirikae

ABSTRACTPseudomonas aeruginosaNCGM1588 has a novel chromosomal class 1 integron, In151, which includes theaac(6′)-Iajgene. The encoded protein, AAC(6′)-Iaj, was found to consist of 184 amino acids, with 70% identity to AAC(6′)-Ia.Escherichia colitransformed with a plasmid containing theaac(6′)-Iajgene acquired resistance to all aminoglycosides tested except gentamicin. Of note,aac(6′)-Iajcontributed to the resistance to arbekacin. Thin-layer chromatography revealed that AAC(6′)-Iaj acetylated all aminoglycosides tested except gentamicin. These findings indicated that AAC(6′)-Iaj is a functional acetyltransferase that modifies the amino groups at the 6′ positions of aminoglycosides and contributes to aminoglycoside resistance ofP. aeruginosaNCGM1588, including arbekacin.


2005 ◽  
Vol 49 (9) ◽  
pp. 3734-3742 ◽  
Author(s):  
Jun-ichiro Sekiguchi ◽  
Tsukasa Asagi ◽  
Tohru Miyoshi-Akiyama ◽  
Tomoko Fujino ◽  
Intetsu Kobayashi ◽  
...  

ABSTRACT We characterized multidrug-resistant Pseudomonas aeruginosa strains isolated from patients involved in an outbreak of catheter-associated urinary tract infections that occurred in a neurosurgery ward of a hospital in Sendai, Japan. Pulsed-field gel electrophoresis of SpeI-, XbaI-, or HpaI-digested genomic DNAs from the isolates revealed that clonal expansion of a P. aeruginosa strain designated IMCJ2.S1 had occurred in the ward. This strain possessed broad-spectrum resistance to aminoglycosides, β-lactams, fluoroquinolones, tetracyclines, sulfonamides, and chlorhexidine. Strain IMCJ2.S1 showed a level of resistance to some kinds of disinfectants similar to that of a control strain of P. aeruginosa, ATCC 27853. IMCJ2.S1 contained a novel class 1 integron, In113, in the chromosome but not on a plasmid. In113 contains an array of three gene cassettes of bla IMP-1, a novel aminoglycoside resistance gene, and the aadA1 gene. The aminoglycoside resistance gene, designated aac(6′)-Iae, encoded a 183-amino-acid protein that shared 57.1% identity with AAC(6′)-Iq. Recombinant AAC(6′)-Iae protein showed aminoglycoside 6′-N-acetyltransferase activity by thin-layer chromatography. Escherichia coli expressing exogenous aac(6′)-Iae showed resistance to amikacin, dibekacin, isepamicin, kanamycin, netilmicin, sisomicin, and tobramycin but not to arbekacin, gentamicins, or streptomycin. Alterations of gyrA and parC at the amino acid sequence level were detected in IMCJ2.S1, suggesting that such mutations confer the resistance to fluoroquinolones observed for this strain. These results indicate that P. aeruginosa IMCJ2.S1 has developed multidrug resistance by acquiring resistance determinants, including a novel member of the aac(6′)-I family and mutations in drug resistance genes.


2008 ◽  
Vol 52 (10) ◽  
pp. 3589-3596 ◽  
Author(s):  
Carlos Juan ◽  
Alejandro Beceiro ◽  
Olivia Gutiérrez ◽  
Sebastián Albertí ◽  
Margalida Garau ◽  
...  

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.


1999 ◽  
Vol 43 (12) ◽  
pp. 2925-2929 ◽  
Author(s):  
Lydia Bass ◽  
Cynthia A. Liebert ◽  
Margie D. Lee ◽  
Anne O. Summers ◽  
David G. White ◽  
...  

ABSTRACT Antibiotic resistance among avian bacterial isolates is common and is of great concern to the poultry industry. Approximately 36% (n = 100) of avian, pathogenic Escherichia coli isolates obtained from diseased poultry exhibited multiple-antibiotic resistance to tetracycline, oxytetracycline, streptomycin, sulfonamides, and gentamicin. Clinical avian E. coli isolates were further screened for the presence of markers for class 1 integrons, the integron recombinase intI1 and the quaternary ammonium resistance gene qacEΔ1, in order to determine the contribution of integrons to the observed multiple-antibiotic resistance phenotypes. Sixty-three percent of the clinical isolates were positive for the class 1 integron markersintI1 and qacEΔ1. PCR analysis with the conserved class 1 integron primers yielded amplicons of approximately 1 kb from E. coli isolates positive for intI1 andqacEΔ1. These PCR amplicons contained the spectinomycin-streptomycin resistance gene aadA1. Further characterization of the identified integrons revealed that many were part of the transposon Tn21, a genetic element that encodes both antibiotic resistance and heavy-metal resistance to mercuric compounds. Fifty percent of the clinical isolates positive for the integron marker gene intI1 as well as for theqacEΔ1 and aadA1 cassettes also contained the mercury reductase gene merA. The correlation between the presence of the merA gene with that of the integrase and antibiotic resistance genes suggests that these integrons are located in Tn21. The presence of these elements among avianE. coli isolates of diverse genetic makeup as well as inSalmonella suggests the mobility of Tn21 among pathogens in humans as well as poultry.


2020 ◽  
Vol 26 (12) ◽  
pp. 1437-1441
Author(s):  
Fatma Zohra Zaidi ◽  
Radia Dali-Yahia ◽  
Karima Zenati ◽  
Leila Yazi ◽  
Manon Lounes ◽  
...  

2017 ◽  
Vol 32 (2) ◽  
pp. 109-115 ◽  
Author(s):  
Hamid Reza Goli ◽  
Mohammad Reza Nahaei ◽  
Mohammad Ahangarzadeh Rezaee ◽  
Alka Hasani ◽  
Hossein Samadi Kafil ◽  
...  

2008 ◽  
Vol 75 (4) ◽  
pp. 1192-1196 ◽  
Author(s):  
Ashraf A. Khan ◽  
Elizabeth Ponce ◽  
M. S. Nawaz ◽  
Chorng-Ming Cheng ◽  
Junaid A. Khan ◽  
...  

ABSTRACT A total of 210 Salmonella isolates, representing 64 different serovars, were isolated from imported seafood samples, and 55/210 isolates were found to be resistant to at least one antibiotic. Class 1 integrons from three multidrug-resistant Salmonella enterica strains (Salmonella enterica serovars Newport [strain 62], Typhimurium var. Copenhagen [strain 629], and Lansing [strain 803], originating from Hong Kong, the Philippines, and Taiwan, respectively) were characterized. Southern hybridization of plasmids isolated from these strains, using a class 1 integron probe, showed that trimethoprim-sulfamethoxazole and streptomycin resistance genes were located on a megaplasmid in strain 629. Our study indicates that imported seafood could be a reservoir for Salmonella isolates resistant to multiple antibiotics.


2019 ◽  
Vol 40 (6) ◽  
pp. 972-978 ◽  
Author(s):  
Siamak Heidarzadeh ◽  
Yasamin Enayati Kaliji ◽  
Reza Pourpaknia ◽  
Alireza Mohammadzadeh ◽  
Mehran Ghazali-Bina ◽  
...  

Abstract The role of integrons has been highlighted in antibiotic resistance among Pseudomonas aeruginosa isolates. Therefore, we here reviewed the prevalence of class 1 integrons and their correlations with antibiotic resistance of P. aeruginosa isolated from Iranian burn patients. This review was conducted according to the guidelines of Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA). Cross-sectional and cohort studies published from January 1, 2000 until December 31, 2018 were enrolled. Meta-analysis was performed by Comprehensive Meta-Analysis (CMA) software using the random effects model, Cochran’s Q, and I2 tests. Publication bias was estimated by Funnel plot and Egger’s linear regression test. Nine out of 819 studies met the eligibility criteria. The overall combined prevalence of class 1 integrons in P. aeruginosa isolates was 69% (95% confidence interval [CI]: 50.5–83%). The highest combined resistance was reported against Cloxacillin (87.7%), followed by Carbenicillin (79.1%) and Ceftriaxone (77.3%). The combined prevalence of multidrug-resistant (MDR) isolates was 79.3% (95% CI: 31.1–97%). Also, a significant correlation was noted between the presence of class 1 integrons and antibiotic resistance in 55.5% of the included studies (P < .05). The results showed high prevalence of class 1 integrons, antibiotic resistance, and MDR strains in P. aeruginosa isolated from Iranian burn patients. Also, most of the included studies showed a significant correlation between the presence of class 1 integrons and antibiotic resistance.


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