J Vet Sci.  2011 Mar;12(1):41-48. 10.4142/jvs.2011.12.1.41.

Molecular characterization of tetracycline- and quinolone-resistant Aeromonas salmonicida isolated in Korea

• Kim JH
• Hwang SY
• Son JS
• Han JE
• Jun JW
• Shin SP
• Choresca C
• Choi YJ
• Park YH
• Park SC

• KMID: 1067336
• DOI: http://doi.org/10.4142/jvs.2011.12.1.41

Abstract

The antibiotic resistance of 16 Aeromonas (A.) salmonicida strains isolated from diseased fish and environmental samples in Korea from 2006 to 2009 were investigated in this study. Tetracycline or quinolone resistance was observed in eight and 16 of the isolates, respectively, based on the measured minimal inhibitory concentrations. Among the tetracycline-resistant strains, seven of the isolates harbored tetA gene and one isolate harbored tetE gene. Additionally, quinolone-resistance determining regions (QRDRs) consisting of the gyrA and parC genes were amplified and sequenced. Among the quinolone-resistant A. salmonicida strains, 15 harbored point mutations in the gyrA codon 83 which were responsible for the corresponding amino acid substitutions of Ser83-->Arg83 or Ser83-->Asn83. We detected no point mutations in other QRDRs, such as gyrA codons 87 and 92, and parC codons 80 and 84. Genetic similarity was assessed via pulsed-field gel electrophoresis, and the results indicated high clonality among the Korean antibiotic-resistant strains of A. salmonicida.

Keyword

Aeromonas salmonicida; minimal inhibitory concentration; pulsed-field gel electrophoresis; quinolone-resistance determining region; tetracycline-resistance

MeSH Terms

Aeromonas salmonicida/classification/*drug effects/*genetics/i
Animals
Anti-Bacterial Agents/*pharmacology
Bacterial Proteins/genetics
*Drug Resistance, Bacterial
Environment
Fish Diseases/*microbiology
Fishes
Gram-Negative Bacterial Infections/microbiology/*veterinary
Microbial Sensitivity Tests
Point Mutation
Polymerase Chain Reaction
Quinolones/*pharmacology
Republic of Korea
Sequence Analysis
Tetracycline/*pharmacology
Tetracycline Resistance

Figure

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  Fig. 1 Multiplex PCR assay of tetracycline resistance genes (tetA of 211 bp and tetE of 744 bp) in two reference strains and 16 isolates of Aeromonas (A.) salmonicida. Lane M: molecular mass marker; lane 1 to 18: strains AS01, AS02, AS03, AS04, AS05, AS06, AS07, AS08, AS09, AS10, AS11, AS12, AS13, AS14, AS15, ASS, ASM, and AS16, respectively. Marker sizes (bp) are indicated.

• 

  Fig. 2 Pulsed-field gel electrophoresis profiles of 18 A. salmonicida strains and UPGMA dendrogram. The vertical dotted line denotes a hypothetical node of 70% Dice coefficient of similarity (2.0% position tolerance).

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