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

Oh-Geun Kweon; Jin Seok Kim; Gou-Ok Kim; Chang-Il Lee; Kwang-Hyeon Jeong; Junyoung Kim

doi:10.5145/ACM.2014.17.2.50

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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

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 bla_CTX-M-27 gene was carried by an IncF conjugative plasmid in the S. Infantis isolate. The bla_CTX-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.

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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..

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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
Antibiotics	Breakpoints		MIC₅₀(μ g/mL)	MIC₉₀(μ g/mL)	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
Strain	Serotype	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
Strain	Serotype	bla _CTX-M gene	Other bla gene	Plasmid replicon	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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