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Kweon, Kim, Kim, Lee, Jeong, and Kim: Characterization of Salmonella spp. Clinical Isolates in Gyeongsangbuk-do Province, 2012 to 2013
Original Article

Ann Clin Microbiol 2014;17(2):50-57.

Published online: 5 June 2014

DOI: https://doi.org/10.5145/ACM.2014.17.2.50

Characterization of Salmonella spp. Clinical Isolates in Gyeongsangbuk-do Province, 2012 to 2013

Oh-Geun Kweon1, Jin Seok Kim2, Gou-Ok Kim1, Chang-Il Lee1, Kwang-Hyeon Jeong1, Junyoung Kim2

1Division of Microbiology, Gyeongsangbuk-do Provincial Government Institute of Health and Environment, Cheongwon, Korea

2Division of Enteric Diseases, Center for Infectious Diseases, Korea National Institute of Health, Cheongwon, Korea

Correspondence: Junyoung Kim, Division of Enteric Diseases, Center for Infectious Diseases, Korea National Institute of Health, Korea Centers for Disease Control and Prevention, 187 Osongsaengmyeong 2(i)-ro, Osong-eup, Cheongwon 363-700, Korea. (Tel) 82-43-719-8116, (Fax) 82-43-719-8149, (E-mail) jun49@hanmail.net

Received 28 February 20148 May 2014       Accepted 14 May 2014

Copyright © 2014 The Korean Society of Clinical Microbiology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

초록

Background

Extended-spectrum cephalosporins and fluoroquinolones are important antimicrobials for treat-ing invasive salmonellosis, and emerging resistance to these antimicrobials is of paramount concern.

Methods

A total of 30 Salmonella spp. clinical isolates recovered in Gyeongsangbuk-do from 2012 to 2013 were characterized using antibiotic resistance profiles and pulsed-field gel electrophoresis (PFGE).

Results

A high prevalence of multidrug-resistant isolates, mainly showing an ampicillin, nalidixic acid, chloramphenicol resistance pattern, was observed. Four extended-spectrum β-lactamase (ESBL)-producing isolates (3 CTX-M-15 isolates and 1 CTX-M-27 iso-late) were found. The blaCTX-M-27 gene was carried by an IncF conjugative plasmid in the S. Infantis isolate. The blaCTX-M-15 gene were carried by an IncF (2 isolates) or IncHI2 (1 isolate) conjugative plasmid in S. Enteritidis. In addition, a single mutation of GyrA, Ser83Thr (1 isolates), Asp87Tyr (9 isolates), Asp87Gly (4 isolates), and Asp87Leu (3 isolates), was detected in nalidixic acid-resistant Salmonella spp. isolates. XbaI PFGE analysis of all isolates revealed more than 19 different pulsotypes. The most common S. Enteritidis PFGE pattern (SEGX01.003) was associated with a larger number of cases of invasive salmonellosis than all other patterns.

Conclusion

The information from our study can as-sist in source attribution, outbreak investigations, and tailoring of interventions to maximize disease prevention.

Keywords: Drug resistance, Extended-spectrum be-ta-lactamase, Nalidixic acid, Pulsed-Field Gel Electrophoresis, Salmonella

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Fig. 1.
Clustering of XbaI digested PFGE patterns for Salmonella spp..
acm-17-50f1.tif
Table 1.
MIC50 and MIC90 of antibiotics for Salmonella spp. isolates (n=30)
Antibiotics Breakpoints MIC50(μ g/mL) MIC90(μ g/mL) No. (%) of isolates
S I R
Ampicillin S≤8 R≥32 128 128 12 (40) 0 18 (60)
Ceftriaxone S≤1 R≥4 0.06 64 26 (86.7) 0 4 (13.3)
Cefotaxime S≤1 R≥4 0.06 8 26 (86.7) 0 4 (13.3)
Cefoxitin S≤8 R≥32 0.5 4 30 (100) 0 0
Ampicillin/Sulbactam S≤8 R≥32 16 32 12 (40) 8 (26.7) 10 (33.3)
Amoxicillin/Clavulanic acid S≤8 R≥32 8 16 25 (83.3) 5 (16.7) 0
Imipenem S≤1 R≥4 0.06 0.06 30 (100) 0 0
Amikacin S≤16 R≥64 0.5 0.5 30 (100) 0 0
Gentamicin S≤4 R≥16 0.5 64 24 (80) 0 6 (20)
Nalidixic acid S≤16 R≥32 256 256 13 (43.3) 0 17 (56.7)
Ciprofloxacin S≤1 R≥4 0.06 0.25 30 (100) 0 0
Tetracycline S≤4 R≥16 0.5 128 23 (76.7) 0 7 (23.3)
Chloramphenicol S≤8 R≥32 8 64 19 (63.3) 0 11 (36.7)
Trimethoprim/Sulfamethoxazole S≤2 R≥4 1 1 28 (93.3) 0 2 (6.7)
Table 2.
MICs of Salmonella spp. isolates
Strain Serotype MICs (μ g/mL) of antibiotics
AMP CRO CTX FOX SAM AMC IMP AMK GEN NAL CIP TCY CHL SXT
1 Typhi 0.5 0.06 0.06 4 1 1 0.06 0.5 0.5 0.5 0.06 0.5 4 1
2 Typhimurium 128 0.06 0.06 0.5 16 (I) 8 0.06 0.5 0.5 256 0.5 256 8 1
3 Sandiego 0.5 0.06 0.06 0.5 1 1 0.06 0.5 0.5 4 0.06 0.5 8 1
4 Enteritidis 128 0.06 0.06 0.05 32 8 0.06 0.5 0.5 256 0.06 0.5 64 1
5 Enteritidis 128 0.06 0.06 0.5 32 8 0.06 0.5 0.5 256 0.25 0.5 64 1
6 Enteritidis 128 64 8 0.5 16 8 0.06 0.5 64 256 0.25 64 4 1
7 Enteritidis 128 0.06 0.06 0.5 16 8 0.06 0.5 0.5 256 0.25 0.5 64 1
8 Senftenberg 0.5 0.06 0.06 4 1 1 0.06 0.5 0.5 4 0.06 0.5 8 1
9 Typhimurium 128 0.06 0.06 0.5 32 8 0.06 0.5 16 4 0.06 128 4 1
10 Enteritidis 128 64 8 0.5 16 8 0.06 0.5 64 256 0.25 128 8 1
11 Typhimurium 0.5 0.06 0.06 0.5 1 1 0.06 0.5 0.5 4 0.06 0.5 8 1
12 Enteritidis 0.5 0.06 0.06 0.5 1 1 0.06 0.5 0.5 256 0.06 0.5 8 1
13 Enteritidis 128 0.06 0.06 0.5 32 16 0.06 0.5 0.5 256 0.06 0.5 64 1
14 Enteritidis 0.5 0.06 0.06 0.5 1 1 0.06 0.5 0.5 256 0.25 0.5 8 1
15 Typhimurium 0.5 0.06 0.06 0.5 2 1 0.06 0.5 0.5 256 0.5 0.5 8 1
16 Enteritidis 128 0.06 0.06 0.5 32 16 0.06 0.5 0.5 256 0.25 0.5 64 1
17 Bardo 0.5 0.06 0.06 0.5 2 2 0.06 0.5 0.5 4 0.06 0.5 8 1
18 Enteritidis 128 0.06 0.06 0.5 32 8 0.06 0.5 0.5 128 0.06 0.5 64 1
19 Thomson 0.5 0.06 0.06 0.5 2 2 0.06 0.5 0.5 4 0.06 0.5 8 1
20 Paratyphi B 128 0.06 0.06 0.5 32 16 0.06 0.5 0.5 0.5 0.06 32 64 1
21 Infantis 128 64 8 4 64 16 0.06 0.5 0.5 4 0.06 0.5 8 32
22 Paratyphi B 128 0.06 0.06 0.5 16 8 0.06 0.5 0.5 256 0.25 0.5 64 1
23 Enteritidis 128 0.06 0.06 0.5 16 8 0.06 0.5 0.5 256 0.06 0.5 64 1
24 Typhimurium 128 0.06 0.06 4 16 8 0.06 0.5 64 256 0.5 4 64 32
25 Typhimurium 0.5 0.06 0.06 4 1 1 0.06 0.5 0.5 4 0.06 0.5 8 1
26 Typhimurium 0.5 0.06 0.06 4 1 1 0.06 0.5 0.5 4 0.06 0.5 8 1
27 Enteritidis 128 0.06 0.06 0.5 32 8 0.06 0.5 0.5 256 0.25 0.5 64 1
28 Enteritidis 128 64 8 0.5 16 4 0.06 0.5 64 256 0.25 128 8 1
29 Stanley 0.5 0.06 0.06 0.5 1 1 0.06 0.5 0.5 0.5 0.06 0.5 8 1
30 Typhimurium 128 0.06 0.06 0.5 64 16 0.06 0.5 128 8 0.06 256 8 1

Abbreviations: AMP, Ampicillin; CRO, Ceftriaxone; CTX, Cefotaxime; FOX, Cefoxitin; SAM, Ampicillin/Sulbactam; AMC, Amoxicillin/ Clavulanic acid; IMP, Imipenem; AMK, Amikacin; GEN, Gentamicin; NAL, Nalidixic acid; CIP, Ciprofloxacin; TCY, Tetracycline; CHL, Chloramphenicol; SXT, Trimethoprim/Sulfamethoxazole. Values in boldface indicate higher value than the breakpoint of CLSI.

Table 3.
Molecular characteristics of Salmonella spp. resistant to third-generation cephalosporins and quinolones
Strain Serotype bla CTX-M gene Other bla gene Plasmid replicon Mutations in QRDR
gyrA parC
2 S. Typhimurium – – – Asp87→ Leu –
4 S. Enteritidis – – – Asp87→ Tyr –
5 S. Enteritidis – – – Asp87→ Tyr –
6 S. Enteritidis CTX-M-15 – IncHI2 Asp87→ Tyr –
7 S. Enteritidis – – – Asp87→ Tyr –
10 S. Enteritidis CTX-M-15 – IncF Asp87→Tyr –
12 S. Enteritidis – – – Asp87→ Gly –
13 S. Enteritidis – – – Asp87→ Gly –
14 S. Enteritidis – – – Asp87→ Tyr –
15 S. Typhimurium – – – Asp87→ Leu –
16 S. Enteritidis – – – Asp87→ Tyr –
18 S. Enteritidis – – – Asp87→Gly –
21 S. Infantis CTX-M-27 TEM-1 IncF – –
22 S. Paratyphi B – – – Ser83→ Thr –
23 S. Enteritidis – – – Asp87→ Gly –
24 S. Typhimurium – – – Asp87→ Leu –
27 S. Enteritidis – – – Asp87→ Tyr –
28 S. Enteritidis CTX-M-15 – IncF Asp87→ Tyr –
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