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Hwang, Song, Kim, Jeong, Kim, and Cheong: Prevalence and Mechanisms of Low Level Quinolone Resistance among Non-Typhoidal Salmonella Isolates from Human and poultry/Livestock in Korea: Usefulness of Nalidixic Acid Resistance Test
Orginal Article

Infect Chemother 2010;42(4):230-236.

Published online: 30 August 2010

DOI: https://doi.org/10.3947/ic.2010.42.4.230

Prevalence and Mechanisms of Low Level Quinolone Resistance among Non-Typhoidal Salmonella Isolates from Human and poultry/Livestock in Korea: Usefulness of Nalidixic Acid Resistance Test

In Sook Hwang1, Joon Young Song1, Woo Joo Kim1, Hye Won Jeong2, Moo Sang Kim3, Hee Jin Cheong1

1Division of Infectious Diseases, Department of Internal Medicine Korea University, College of Medicine, Seoul, Korea

2Division of Internal Medicine, Department of Medicine Chungbuk National University, Cheongju, Korea

3Seoul Metropolitan Government Research Institute of Public Health and Environment, Seoul, Korea

Correspondence to Hee-Jin Cheong, M.D., Ph.D. Division of Infectious Disease, Department of Internal Medicine, Korea University College of Medicine, 97 Guro Dong-gil, Guro-gu, Seoul 152-703, Korea Tel: +82-2-2626-3050, Fax: +82-2-2626-1105 E-mail: heejinmd@medimail.co.kr

This study was supported by Korea University BK21 (K0518051) grants.

Received 11 June 2010       Revised 9 August 2010       Accepted 9 August 2010

Copyright © 2010 The Korean Society of Infectious Diseases; Korean Society for Chemotherapy

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.

Abstract

Background

Non-typhoidal Salmonella (NTS) are important commensal microorganisms. We intended to investigate the prevalence and mechanisms of nalidixic acid resistance among NTS isolated from human and poultry/livestock.

Methods

A total of 151 Salmonella isolates (36 human and 115 livestock isolates, respectively) was tested for the Minimum inhibitory concentrations (MICs) of nalidixic acid, together with serotyping. As for the nalidixic acid resistant isolates, further studies were taken: MICs of ciprofloxacin, mutation analysis of gyrA and parC genes, and organic solvent tolerance test.

Results

Eighty-four isolates of 151 human and livestock isolates were resistant to nalidixic acid. The prevalence of nalidixic acid resistance and was 13.9% (5 of 36 isolates) in human isolates and 68.7% (79 of 151 isolates), in the livestock isolates respectively. Among 84 nalidixic acid-resistant isolates, the The prevalence of ciprofloxacin resistance in livestock isolates was 24.1% (1 resistant and 18 intermediate of 79 strains), but no ciprofloxacin resistance was found in 5 human isolates. Among 65 nalidixic acid resistant, ciprofloxacin-susceptible isolates, 3 (60%, of 5 human isolates) and 60 (100%, all livestock isolates) showed low level fluoroquinolone resistance (ciprofloxacin MIC, 0.125-1.0 μg/μL). Six types of point mutations were found in the analysis of DNA sequencing of the gyrA gene in the 84 isolates; 75 isolates showed point mutations on amino acid Ser 83 and/or Asp 87. On the other hand, no point mutation was found from the parC genes. Forty-seven nalidixic acid resistant isolates showed tolerance to organic solvents.

Conclusions

Nalidixic acid resistance was a good marker of low level fluoroquinolone resistance. As for the severe NTS infection, MIC test for nalidixic acid would be required.

Keywords: Non-typhoidal Salmonella, Nalidixic acid, Low level fluoroquinolone resistance

REFERENCES

1. San Martin B, Lapierre L, Toro C, Bravo V, Cornejo J, Hormazabal JC, Borie C. Isolation and molecular characterization of quinolone resistant Salmonella spp. from poultry farms. Vet Microbiol. 2005; 110:239–244.
2. Marimón JM, Gomáriz M, Zigorraga C, Cilla G, Pérez-Trallero E. Increasing prevalence of quinolone resistance in human nontyphoid Salmonella enterica isolates obtained in Spain from 1981 to 2003. Antimicrob Agents Chemother. 2004; 48:3789–3793.
3. Hakanen A, Kotilainen P, Jalava J, Siitonen A, Huovinen P. Detection of decreased fluoroquinolone susceptibility in Salmonellas and validation of nalidixic acid screening test. J Clin Microbiol. 1999; 37:3572–3577.
4. Breuil J, Brisabois A, Casin I, Armand-Lefèvre L, Frémy S, Collatz E. Antibiotic resistance in salmonellae isolated from humans and animals in France: comparative data from 1994 and 1997. J Antimicrob Chemother. 2000; 46:965–971.
crossref
5. Hakanen A, Siitonen A, Kotilainen P, Huovinen P. Increasing fluoroquinolone resistance in Salmonella serotypes in Finland during 1995-1997. J Antimicrob Chemother. 1999; 43:145–148.
6. Herikstad H, Hayes P, Mokhtar M, Fracaro ML, Threlfall EJ, Angulo FJ. Emerging quinolone-resistant Salmonella in the United States. Emerg Infect Dis. 1997; 3:371–372.
7. Mølbak K, Gerner-Smidt P, Wegener HC. Increasing quinolone resistance in Salmonella enterica serotype Enteritidis. Emerg Infect Dis. 2002; 8:514–515.
8. Prats G, Mirelis B, Llovet T, Muñoz C, Miró E, Navarro F. Antibiotic resistance trends in enteropathogenic bacteria isolated in 1985-1987 and 1995-1998 in Barcelona. Antimicrob Agents Chemother. 2000; 44:1140–1145.
crossref
9. Seral C, López L, Castillo FJ, Clavel A, Rubio MC. Quinolone resistance in Salmonella enterica. Rev Esp Quimioter. 1998; 11:43–46.
10. Threlfall EJ, Ward LR, Rowe B. Resistance to ciprofloxacin in non-typhoidal salmonellas from humans in England and Wales-the current situation. Clin Microbiol Infect. 1999; 5:130–134.
11. Walker RA, Saunders N, Lawson AJ, Lindsay EA, Dassama M, Ward LR, Woodward MJ, Davies RH, Liebana E, Threlfall EJ. Use of a LightCycler gyrA mutation assay for rapid identification of mutations conferring decreased susceptibility to ciprofloxacin in multiresistant Salmonella enterica serotype Typhimurium DT104 isolates. J Clin Microbiol. 2001; 39:1443–1448.
12. Hirose K, Hashimoto A, Tamura K, Kawamura Y, Ezaki T, Sagara H, Watanabe H. DNA sequence analysis of DNA gyrase and DNA topoisomerase IV quinolone resistance-determining regions of Salmonella enterica serovar Typhi and serovar Paratyphi A. Antimicrob Agents Chemother. 2002; 46:3249–3252.
13. Liebana E, Clouting C, Cassar CA, Randall LP, Walker RA, Threlfall EJ, Clifton-Hadley FA, Ridley AM, Davies RH. Comparison of gyrA mutations, cyclohexane resistance, and the presence of class I integrons in Salmonella enterica from farm animals in England and Wales. J Clin Microbiol. 2002; 40:1481–1486.
14. Reche MP, García de los Ríos JE, Jiménez PA, Rojas AM, Rotger R. gyrA Mutations associated with nalidixic acid-resistant salmonellae from wild birds. Antimicrob Agents Chemother. 2002; 46:3108–3109.
15. Clinical and Laboratory Standards Institute. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. 7th ed. Wayne, PA: 2006. Document M7-A7.
16. Popoff MY, Bockemühl J, Brenner FW. Supplement 1997 (no. 41) to the Kauffmann-White scheme. Res Microbiol. 1998; 149:601–604.
crossref
17. Asako H, Nakajima H, Kobayashi K, Kobayashi M, Aono R. Organic solvent tolerance and antibiotic resistance increased by overexpression of marA in Escherichia coli. Appl Environ Microbiol. 1997; 63:1428–1433.
18. Oethinger M, Kern WV, Goldman JD, Levy SB. Association of organic solvent tolerance and fluoroquinolone resistance in clinical isolates of Escherichia coli. J Antimicrob Chemother. 1998; 41:111–114.
19. Threlfall EJ. Antimicrobial drug resistance in Salmonella: problems and perspectives in food- and water-borne infections. FEMS Microbiol Rev. 2002; 26:141–148.
20. Hakanen AJ, Kotilainen P, Pitkänen S, Huikko S, Siitonen A, Huovinen P. Reduction in fluoroquinolone susceptibility among non-typhoidal strains of Salmonella enterica isolated from Finnish patients. J Antimicrob Chemother. 2006; 57:569–572.
21. Murray A, Coia JE, Mather H, Brown DJ. Ciprofloxacin resistance in non-typhoidal Salmonella serotypes in Scotland, 1993-2003. J Antimicrob Chemother. 2005; 56:110–114.
22. Chang CM, Lee HC, Lee NY, Huang GC, Lee IW, Ko WC. Cefotaxime-ciprofloxacin combination therapy for nontyphoid Salmonella bacteremia and paravertebral abscess after failure of monotherapy. Pharmacotherapy. 2006; 26:1671–1674.
23. Heisig P. High-level fluoroquinolone resistance in a Salmonella Typhimurium isolate due to alterations in both gyrA and gyrB genes. J Antimicrob Chemother. 1993; 32:367–377.
24. Bauernfeind A, Casellas JM, Goldberg M, Holley M, Jungwirth R, Mangold P, Röhnisch T, Schweighart S, Wilhelm R. A new plasmidic cefotaximase from patients infected with Salmonella Typhimurium. Infection. 1992; 20:158–163.
25. Digranes A, Solberg CO, Sjursen H, Skovlund E, Sander J. Antibiotic susceptibility of blood culture isolates of Enterobacteriaceae from six Norwegian hospitals 1991-1992. APMIS. 1997; 105:854–860.
crossref
26. Chung TH, Yun SK, Lee BK, Choi JD, Lee MW. Study on the Genus Salmonella cultures isolated in Korea 1982. J Korean Soc Microbiol. 1983; 18:31–38.
27. Stevenson JE, Gay K, Barrett TJ, Medalla F, Chiller TM, Angulo FJ. Increase in nalidixic acid resistance among non-Typhi Salmonella enterica isolates in the United States from 1996 to 2003. Antimicrob Agents Chemother. 2007; 51:195–197.
28. Choi SH, Woo JH, Lee JE, Park SJ, Choo EJ, Kwak YG, Kim MN, Choi MS, Lee NY, Lee BK, Kim NJ, Jeong JY, Ryu J, Kim YS. Increasing incidence of quinolone resistance in human non-typhoid Salmonella enterica isolates in Korea and mechanisms involved in quinolone resistance. J Antimicrob Chemother. 2005; 56:1111–1114.
29. Cheong HJ, Lee YJ, Hwang IS, Kee SY, Cheong HW, Song JY, Kim JM, Park YH, Jung JH, Kim WJ. Characteristics of non-typhoidal Salmonella isolates from human and broiler-chickens in southwestern Seoul, Korea. J Korean Med Sci. 2007; 22:773–778.
30. Helms M, Vastrup P, Gerner-Smidt P, Mølbak K. Excess mortality associated with antimicrobial drug-resistant Salmonella Typhimurium. Emerg Infect Dis. 2002; 8:490–495.
31. Asna SMZ, Haq JA, Rahaman MM. Nalidixic acid-resistant Salmonella enterica serovar Typhi with decreased susceptibility to ciprofloxacin caused treatment failure: a report from Bangladesh. Jpn J Infect Dis. 2003; 56:32–33.
32. Aarestrup FM, Wiuff C, Mølbak K, Threlfall EJ. Is it time to change fluoroquinolone breakpoints for Salmonella spp.? Antimicrob Agents Chemother. 2003; 47:827–829.
33. Parry CM. Antimicrobial drug resistance in Salmonella enterica. Curr Opin Infect Dis. 2003; 16:467–472.
34. Eaves DJ, Randall L, Gray DT, Buckley A, Woodward MJ, White AP, Piddock LJ. Prevalence of mutations within the quinolone resistance-determining region of gyrA, gyrB, parC, and parE and association with antibiotic resistance in quinolone-resistant Salmonella enterica. Antimicrob Agents Chemother. 2004; 48:4012–4015.
35. Mølbak K, Baggesen DL, Aarestrup FM, Ebbesen JM, Engberg J, Frydendahl K, Gerner-Smidt P, Petersen AM, Wegener HC. An outbreak of multidrug-resistant, quinolone-resistant Salmonella enterica serotype Typhimurium DT104. N Engl J Med. 1999; 341:1420–1425.

Table 1.
Serotype Distribution of Human and Livestock among Non-Typhoidal Salmonella Isolates
Serotypes No. of human isolate (%) No. of livestock isolate (%)
Salmonella Enteritidis 17 (47.2) 15 (13.1)
Salmonella Typhimurium 8 (22.2) 16 (13.9)
Salmonella Brandenburg 1 (2.8)
Salmonella Heidelberg 1 (2.8)
Salmonella Istanbul 1 (2.8)
Salmonella Kambole 1 (2.8)
Salmonella London 1 (2.8)
Salmonella Montevideo 1 (2.8) 6 (5.2)
Salmonella Newport 1 (2.8)
Salmonella Ohio 1 (2.8)
Salmonella Rissen 1 (2.8)
Salmonella Schwarzengrund 1 (2.8)
Salmonella Tennessee 1 (2.8) 13 (11.3)
Salmonella Bovismorbificans 3 (2.6)
Salmonella Bsilla 2 (1.7)
Salmonella Kisii 1 (0.9)
Salmonella Salamae 9 (7.8)
Salmonella Virginia 4 (3.5)
Salmonella Yovokome 43 (37.4)
Non-identifed 3 (2.6)
Total 36 (100) 115 (100)
Table 2.
Ciprofloxacin MIC of the Nalidixic Acid-Resistant Non-Typhoidal Salmonella Isolates
Human isolates (n=5/36, 13.9%) Livestock isolates (n=79/115, 68.7%)
No. of nalidixic acid resistant isolates (n=84) 5 79
MIC range (μg/mL) 32 - >128 128 - >128
Classification based on ciprofloxacin S I R S I R
 No. (%) 5 (100) 0 (0) 0 (0) 60 (75.9) 18 (22.8) 1 (1.3)
 MIC range (μg/mL) 0.06 - 1 - - 0.125 - 1 2 4
 No. of low level quinolone resistance (%) 3 (60) - - 60 (100) - -

MIC, Minimal inhibitory concentration; S, Susceptible; I, Intermediate; R, Resistant

Ciprofloxacin MIC value: Susceptible (≤1 μg/mL), Intermediate (2 μg/mL), Resistance (≥4 μg/mL).

Nalidixic acid MIC value: Susceptible (≤16 μg/mL), Intermediate (-), Resistance (≥32 μg/mL).

Low level quinolone resistance MIC range 0.125 to 1.0 μg/mL.

Table 3.
Sequence of a Portion of QRDR in gyrA and parC of the Nalidixic Acid-Resistant Non-Typhoidal Salmonella
gyrA mutation
 parC mutation
 No. of isolates (%) MIC range (μg/mL)
Codon 83 Codon 87 Codon 57, 66, 80 NA CIP
Ser83 (TCG) → Phe (TTC) Wild type Wild type 59 (70.2) 128 - >128 0.125 - 4
Ser83 (TCG) → Phe (TTC or TAC) Wild type Wild type 5 (6.0) >128 0.5 - 2
Ser83 (TCG) → Phe (TTC or TAC) Asp87 (GAC) → Glu (GAA or GAG) Wild type 6 (7.1) 128 - >128 0.25 - 2
Ser83 (TCG) → Phe (TTC) Asp87 (GAC) → Arg (CGA) Wild type 1 (1.2) >128 2
Ser83 (TCG) → Phe (TTC) Asp87 (GAC) → Tyr (TAC) Wild type 2 (2.4) 128 1
Ser83 (TCG) → ? Wild type Wild type. 1 (1.2) 128 1
Wild type Asp87 (GAC) → Asn (AAC) Wild type 1 (1.2) >128 0.06
Wild type Wild type Wild type. 9 (10.7) 32 - >128 0.06 - 1

A total of 84 isolates was examined.

CIP, Ciprofloxacin; NA, Nalidixic acid

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