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Yum, Shin, Yong, and Chong: Diversity of Integrons Carrying blaVIM-2 Cassette in Pseudomonas spp. and Acinetobacter spp.
Original Article
Korean Journal of Clinical Microbiology

Korean Journal of Clinical Microbiology 2012; 15(4): 131-138.

Published online: 21 December 2012

DOI: https://doi.org/10.5145/KJCM.2012.15.4.131

Diversity of Integrons Carrying blaVIM-2 Cassette in Pseudomonas spp. and Acinetobacter spp.

Jonghwa Yum1, Hee Bong Shin2, Dongeun Yong3, Yunsop Chong3

1Department of Clinical Laboratory Science, College of Nursing and Healthcare Sciences, Dong-eui University, Busan, Korea.

2Department of Laboratory Medicine, Soonchunhyang University Hospital, Bucheon, Korea.

3Department of Laboratory Medicine and Research Institute of Bacterial Resistance, College of Medicine, Yonsei University, Seoul, Korea.

Correspondence: Jonghwa Yum, Department of Clinical Laboratory Science, College of Nursing and Healthcare Sciences, Dong-eui University, 995, Eomgwang-ro, Busanjin-gu, Busan 614-714, Korea. (Tel) 82-51-890-2682, (Fax) 82-51-890-1469, auxotype@deu.ac.kr

Received 6 August 2012       Revised 6 October 2012       Accepted 8 October 2012

Copyright © 2012 The Korean Society of Clinical Microbiology

Abstract

Background

Metallo-β-lactamase-mediated carbapenem resistance has been increasingly reported in Pseudomonas, Acinetobacter and other Gram-negative bacilli (GNB) in many countries. A few studies showed highly variable structure of MBL-gene cassette-carrying integrons. The aim of this study was to determine the structure of blaVIM-2-carrying integrons in Pseudomonas and Acinetobacter.

Methods

blaVIM-2-carrying GNB were isolated at a Korean hospitals during the years 1995-1999 and 2005. The size of blaVIM-2-carrying integrons was estimated by the PCR products. Representative integrons were sequenced by the dideoxy-chain termination method. The MICs of antimicrobial agents were tested by the CLSI agar dilution methods.

Results

During the years 1995-1999 and 2005, the approximate size of the blaVIM-2-carrying class 1 integrons was 3-7 kb in 35 Pseudomonas isolates and 3-5 kb in 24 Acinetobacter isolates. The integrons carried one-five resistance gene cassettes in addition to the blaVIM-2 cassette. Other resistance gene cassettes found were blaOXA-1, aacA1, aac(6')-I, and aac(6')-II. Interestingly, sequences homologous to part of a putative class II intron were inserted into the recombination site of the last cassette in four of nine integrons. The class 1 integron from P. aeruginosa isolates had fused orf/IntI1 in a downstream leftward inverted repeat (IRi).

Conclusion

According to period, the size and structure of blaVIM-2-carrying integrons are quite variable, but an identical one is also present in a different genus, indicating high mobility of the blaVIM-2 cassette and horizontal transfer of the whole integron. We suggest that the class 1 integron containing the blaVIM-2 gene is spreading horizontally among Gram-negative bacilli and is undergoing continuous development in Korea.

Keywords: Gram-negative bacilli, Integron, Metallo-β-lactamase

Figures and Tables

Fig. 1
Comparison of the structures of variable regions of class 1 integrons determined in this study. Hatched rectangles indicate the 5'- and 3'-conserved segments (5'-CS and 3'-CS), black arrows the cassette-borne resistance genes, empty rectangles the attC sites of the cassettes, small rectangle the IRi sites of the integron, and gray circles the recombination core sites and inverse core sites. The insertion and orientation of part of putative class II intron in the recombination site of the last cassette of In106 and integron in P. putida YMC 99/10/431, P. aeruginosa YMC 95/1/704, and P. aeruginosa YMC 05/5/1103 are indicated.
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Fig. 2
Deduced amino acid changes of aac(6')-I detected in this study, compared to aac(6')-I deduced amino acid reported by Nobuta et al. [10], P. aeruginosa YMC 95/1/2105 and P. putida YMC 99/10/431 had a mutation T269C and deduced amino acid change of L90S. P. aeruginosa YMC 95/1/2105 also had G298A and T512A mutations, resulting in amino acid changes of G100S and V171D. Isolates with mutations in this study: Study No. 3, 6 in Table 2. P. aeruginosa YMC 95/1/704 and P. putida YMC 99/10/431; No. 2. Acinetobacter genomospecies 3 YMC 99/11/U160; No. 4. P. aeruginosa YMC 95/1/2105.
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Table 1
Charteristics of P. aeruginosa, P. putida and Acinetoabcter isolates with blaVIM-2 carrying integrons
kjcm-15-131-i001

Abbreviations: STR, streptomycin; SPT, spectinomycin; KAN, kanamycin; AMK, amikacin; GEN, gentamicin; ISP, isepamicin; NET, netilmicin; TOB, tobramycin; S, susceptible; I, intermediate; R, resistant; NT, not tested because probably due to presence of the gene in other genetic elements.

Table 2
MIC of antimicrobial agents for blaVIM-2-positive gram-negative bacilli with different position of blaVIM-2 cassette in class 1 integrons
kjcm-15-131-i002

Abbreviations: PIP, piperacillin; CTX, cefotaxime; CAZ, ceftazidime; ATM, aztreonam; IPM, imipenem; MEM, meropenem.

Table 3
Amino acid substitutions in aac(6')-I type aminoglycoside modifying enzymes
kjcm-15-131-i003

Abbreiations: Leu, leucine; Gly, glycine; Val, valine; Ser, serine; Asp, asparagine.

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