Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Characterization of Drug-Resistant Salmonella enterica Serotype Typhimurium by Antibiograms, Plasmids, Integrons, Resistance Genes, and PFGE

  • Benacer, Douadi (Institute of Biological Sciences, Faculty of Science, University of Malaya) ;
  • Thong, Kwai Lin (Institute of Biological Sciences, Faculty of Science, University of Malaya) ;
  • Watanabe, Haruo (National Institute of Infectious Disease) ;
  • Puthucheary, Savithri Devi (Department of Medical Microbiology, Faculty of Medicine, University of Malaya)
  • Received : 2009.10.21
  • Accepted : 2010.02.18
  • Published : 2010.06.28
Download PDF
⟨ Previous Next ⟩

Abstract

Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4% of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2%), sulfonamides (57.4%), streptomycin (53.1%), ampicillin (29.7%), nalidixic acid (27.6%), kanamycin (23.4%), chloramphenicol (21.2%), and trimethoprim (19.1%). Resistance towards cephalosporins was noted for cephalothin (27.6%), cephradine (21.2%), amoxicillin clavulanic acid (17.0%), and cephalexin (17.0%). Resistance genes, $bla_{TEM}$, strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirtythree strains (70.2%) carried class 1 integrons, which were grouped in 9 different profiles. DNA sequencing identified sat, aadA, pse-1, and dfrA genes in variable regions on class 1 integrons. Thirty-five strains (74.4%) were subtyped to 22 different plasmid profiles, each with 1-6 plasmids (2.0 to 95 kb). PFGE subtyped the 47 strains into 39 profiles. In conclusion, high rates of multidrug resistance were found among the Malaysian Salmonella Typhimurium strains. The emergence of multidrug-resistant Salmonella Typhimurium to cephalosporin antibiotics was also observed. The strains were very diverse and no persistent clone was observed. The emergence of MDR Salmonella Typhimurium is a worldwide problem, and this report provides information for the better understanding of the prevalence and epidemiology of MDR S. Typhimurium in Malaysia.

Keywords

References

  1. Aarestrup, F. M., C. Wiuff, K. Molbak, and E. J. Threlfall. 2003. Is it time to change fluoroquinolone breakpoints for Salmonella spp.? Antimicrob. Agents Chemother. 47: 827-829. https://doi.org/10.1128/AAC.47.2.827-829.2003
  2. Aarestrup, F. M., M. Lertworapreecha, M. C. Evans, A. Bangtrakulnonth, T. Chalermchaikit, R. S. Hendriksen, and H. C. Wegener. 2003. Antimicrobial susceptibility and occurrence of resistance genes among Salmonella enterica serovar Weltevreden from different countries. J. Antimicrob. Chemother. 52: 715-718. https://doi.org/10.1093/jac/dkg426
  3. Adaska, J. M., A. J. Silva, A. C. Berge, and W. M. Sischo. 2006. Genetic and phenotypic variability among Salmonella enterica serovar Typhimurium isolates from California dairy cattle and humans. Appl. Environ. Microbiol. 72: 6632-6637. https://doi.org/10.1128/AEM.01038-06
  4. Alvarez, J., M. Sota, A. B. Vivanco, I. Perales, R. Cisterna, A. Rementeria, and J. Garaizar. 2004. Development of a multiplex PCR technique for detection and epidemiological typing of Salmonella in human clinical samples. J. Clin. Microbiol. 42: 1734-1738. https://doi.org/10.1128/JCM.42.4.1734-1738.2004
  5. Antunes, P., J. Machado, J. C. Sousa, and L. Peixe. 2005. Dissemination of sulfonamide resistance genes (sul1, sul2, and sul3) in Portuguese Salmonella enterica strains and relation with integrons. Antimicrob. Agents Chemother. 49: 836-839. https://doi.org/10.1128/AAC.49.2.836-839.2005
  6. Antunes, P., J. Machado, and L. Peixe. 2006. Characterization of antimicrobial resistance and class 1 and 2 integrons in Salmonella enterica isolates from different sources in Portugal. J. Antimicrob. Chemother. 58: 297-304. https://doi.org/10.1093/jac/dkl242
  7. Biendo, M., G. Laurans, D. Thomas, B. Canarelli, F. Hamdad-Daoudi, F. Rousseau, S. Castelain, and F. Eb. 2005. Molecular characterisation and mechanisms of resistance of multidrug-resistant human Salmonella enterica serovar Typhimurium isolated in Amiens (France). Int. J. Antimicrob. Agents 26: 219-229. https://doi.org/10.1016/j.ijantimicag.2005.05.003
  8. Boyd, D. A., G. A. Peters, L.-K. Ng, and M. R. Mulvey. 2000. Partial characterization of a genomic island associated with the multidrug resistance region of Salmonella enterica Typhimurium DT104. FEMS Microbiol. Lett. 189: 285-291. https://doi.org/10.1111/j.1574-6968.2000.tb09245.x
  9. Cardoso, M. O., A. R. Ribeiro, L. R. dos Santos, F. Pilotto, H. L. S. de Moraes, C. T. P. Salle, S. L. S. Rocha, and V. P. do Nascimento. 2006. Antibiotic resistance in Salmonella Enteritidis isolated from broiler carcasses. Braz. J. Microbiol. 37: 368-371. https://doi.org/10.1590/S1517-83822006000300030
  10. Chen, S., S. Zhao, D. G. White, C. M. Schroeder, R. Lu, H. Yang, P. F. McDermott, S. Ayers, and J. Meng. 2004. Characterization of multiple-antimicrobial-resistant Salmonella serovars isolated from retail meats. Appl. Environ. Microbiol. 70: 1-7. https://doi.org/10.1128/AEM.70.1.1-7.2004
  11. Clinical and Laboratory Standards Institute. 2005. Performance Standards for Antimicrobial Susceptibility Testing Approved Standard. M100-S15. Clinical and Laboratory Standards Institute, Wayne, PA.
  12. Daly, M., J. Buckley, E. Power, C. O'Hare, M. Cormican, B. Cryan, P. G. Wall, and S. Fanning. 2000. Molecular characterization of Irish Salmonella enterica serotype Typhimurium: Detection of class I integrons and assessment of genetic relationships by DNA amplification fingerprinting. Appl. Environ. Microbiol. 66: 614-619. https://doi.org/10.1128/AEM.66.2.614-619.2000
  13. Foley, S. L., D. G. White, P. F. McDermott, R. D. Walker, B. Rhodes, P. J. Fedorka-Cray, S. Simjee, and S. Zhao. 2006. Comparison of subtyping methods for differentiating Salmonella enterica serovar typhimurium isolates obtained from food animal sources. J. Clin. Microbiol. 44: 3569-3577. https://doi.org/10.1128/JCM.00745-06
  14. Guerra, B., S. Soto, S. Cal, and M. C. Mendoza. 2000. Antimicrobial resistance and spread of class 1 integrons among Salmonella serotypes. Antimicrob. Agents Chemother. 44: 2166-2169. https://doi.org/10.1128/AAC.44.8.2166-2169.2000
  15. Guerra, B., S. M. Soto, J. M. Argüelles, and M. C. Mendoza. 2001. Multidrug resistance is mediated by large plasmids carrying a class 1 integron in the emergent Salmonella enterica serotype [4,5,12:i:-]. Antimicrob. Agents Chemother. 45: 1305-1308. https://doi.org/10.1128/AAC.45.4.1305-1308.2001
  16. Hansen, J. B. and R. H. Olsen. 1978. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J. Bacteriol. 135: 227-238.
  17. Heurtin-Le Corre, C., P. Y. Donnio, M. Perrin, M. F. Travert, and J. L. Avril. 1999. Increasing incidence and comparison of nalidixic acid-resistant Salmonella enterica subsp. enterica serotype Typhimurium isolates from humans and animals. J. Clin. Microbiol. 37: 266-269.
  18. Lailler, R., F. Grimont, Y. Jones, P. Sanders, and A. Brisabois. 2002. Subtyping of Salmonella Typhimurium by pulsed-field gel electrophoresis and comparisons with phage types and resistance types. Pathol. Biol. 50: 361-368. https://doi.org/10.1016/S0369-8114(02)00321-8
  19. Lauderdale, T. L., F. M. Aarestrup, P. C. Chen, J. F. Lai, H. Y. Wang, Y. R. Shiau, I. W. Huang, C. L. Hung, and TSAR hospitals. 2006. Multidrug resistance among different serotypes of clinical Salmonella isolates in Taiwan. Diagn. Microbiol. Infect. Dis. 55: 149-155. https://doi.org/10.1016/j.diagmicrobio.2006.01.002
  20. Levesque, C., L. Pyche, C. Larose, and P. H. Roy. 1995. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob. Agents Chemother. 39: 185-191. https://doi.org/10.1128/AAC.39.1.185
  21. Liebana, E., L. Garcia-Migura, C. Clouting, F. A. Clifton-Hadley, E. Lindsay, E. J. Threlfall, S. W. McDowell, and R. H. Davies. 2002. Multiple genetic typing of Salmonella enterica serotype Typhimurium isolates of different phage types (DT104, U302, DT204b, and DT49) from animals and humans in England, Wales, and Northern Ireland. J. Clin. Microbiol. 40: 4450-4456. https://doi.org/10.1128/JCM.40.12.4450-4456.2002
  22. Ng, L. K., M. R. Mulvey, I. Martin, G. A. Peters, and W. Johnson. 1999. Genetic characterization of antimicrobial resistance in Canadian isolates of Salmonella serovar Typhimurium DT104. Antimicrob. Agents Chemother. 43: 3018-3021.
  23. Oliver, A., L. M. Weigel, J. K. Rasheed, J. E. McGowan, P. Raney, and F. C. Tenover. 2002. Mechanisms of decreased susceptibility to cefpodoxime in Escherichia coli. Antimicrob. Agents Chemother. 46: 3829-3836. https://doi.org/10.1128/AAC.46.12.3829-3836.2002
  24. Olsen, J. E., D. J. Brown, D. L. Baggesen, and M. Bisgaard. 1994. Stability of plasmids in five strains of Salmonella maintained in stab culture at different temperatures. J. Appl. Bacteriol. 77: 155-159. https://doi.org/10.1111/j.1365-2672.1994.tb03059.x
  25. Randall, L. P., S. W. Cooles, M. K. Osborn, L. J. Piddock, and M. J. Woodward. 2004. Antibiotic resistance genes, integrons and multiple antibiotic resistance in thirty-five serotypes of Salmonella enterica isolated from humans and animals in the U.K. J. Antimicrob. Chemother. 53: 208-216. https://doi.org/10.1093/jac/dkh070
  26. Tassios, P. T., M. Gazouli, E. Tzelepi, H. Milch, N. Kozlova, S. Sidorenko, N. J. Legakis, and L. S. Tzouvelekis. 1999. Spread of a Salmonella typhimurium clone resistant to expanded-spectrum cephalosporins in three European countries. J. Clin. Microbiol. 37: 3774-3777.
  27. Taylor, D. N., I. K. Wachsmuth, Y. H. Shangkuan, E. V. Schmidt, T. J. Barrett, J. S. Shchrader, et al. 1982. Salmonellosis associated with marijuana: A multistate outbreak traced by plasmid fingerprinting. N. Engl. J. Med. 306: 1249-1253. https://doi.org/10.1056/NEJM198205273062101
  28. Thong, K. L., K. S. Lai, and Y. L. Goh. 2003. Technical improvement in DNA preparation in pulsed field electrophoresis. Malaysian J. Sci. 22: 55-59.
  29. Threlfall, E. J. 2000. Multiresistant Salmonella Typhimurium DT 104: A truly international multiresistant clone. J. Antimicrob. Chemother. 46: 7-10. https://doi.org/10.1093/jac/46.1.7
  30. Threlfall, E. J. 2002. Antimicrobial drug resistance in Salmonella: Problems and perspectives in food- and water-borne infections. FEMS Microbiol. Rev. 26: 141-148. https://doi.org/10.1111/j.1574-6976.2002.tb00606.x
  31. Wain, J., L. T. Diem Nga, C. Kidgell, K. James, S. Fortune, T. Song Diep, et al. 2003. Molecular analysis of incHI1 antimicrobial resistance plasmids from Salmonella serovar Typhi strains associated with typhoid fever. Antimicrob. Agents Chemother. 47: 2732-2739. https://doi.org/10.1128/AAC.47.9.2732-2739.2003
  32. Woo, Y. K. and S. H. Lee. 2006. Genetic diversity of multiresistant Salmonella enterica serotype Typhimurium isolates from animals and humans. J. Microbiol. 44: 106-112.
  33. Yang, S. J., K.Y. Park, S. H. Kim, K. M. No, T. E. Besser, H. S. Yoo, S. H. Kim, B. K. Lee, and Y. H. Park. 2002. Antimicrobial resistance in Salmonella enterica serovars Enteritidis and Typhimurium isolated from animals in Korea: Comparison of phenotypic and genotypic resistance characterization. Vet. Microbiol. 86: 295-301. https://doi.org/10.1016/S0378-1135(02)00009-3
  34. Yasin, R. M., M. M. Jegathesan, and C. C. Tiew. 1997. Salmonella serotypes isolated in Malaysia over the ten-year period 1983-1992. Asia Pac. J. Public Health 9: 1-5. https://doi.org/10.1177/101053959700900101

Cited by

  1. Molecular and comparative analysis of Salmonella enterica Senftenberg from humans and animals using PFGE, MLST and NARMS vol.11, pp.None, 2011, https://doi.org/10.1186/1471-2180-11-153
  2. Antimicrobial Resistance inSalmonellaSerovars Isolated from Meat Shops at the Markets in North Vietnam vol.9, pp.11, 2010, https://doi.org/10.1089/fpd.2011.1121
  3. High prevalence of multidrug resistance in bacterial uropathogens from Kathmandu, Nepal vol.5, pp.None, 2010, https://doi.org/10.1186/1756-0500-5-38
  4. Molecular Characterization of Antibiotic-Resistant Salmonella Isolates from Retail Meat from Markets in Northern Vietnam vol.75, pp.9, 2010, https://doi.org/10.4315/0362-028x.12-101
  5. The antibiotic resistance characteristics of non-typhoidal Salmonella enterica isolated from food-producing animals, retail meat and humans in South East Asia vol.154, pp.3, 2010, https://doi.org/10.1016/j.ijfoodmicro.2011.12.032
  6. Isolation and Determination of Antibiotic Resistance Patterns in Non-typhoid Salmonella spp isolated from chicken vol.1, pp.1, 2010, https://doi.org/10.17795/ijep9416
  7. Resistance Genes, Phage Types and Pulsed Field Gel Electrophoresis Pulsotypes in Salmonella enterica Strains from Laying Hen Farms in Southern Italy vol.10, pp.8, 2010, https://doi.org/10.3390/ijerph10083347
  8. High Resolution Melting Analysis for Rapid Mutation Screening in Gyrase and Topoisomerase IV Genes in Quinolone-Resistant Salmonella enterica vol.2014, pp.None, 2010, https://doi.org/10.1155/2014/718084
  9. Investigation of Class I Integron in Salmonella infantis and Its Association With Drug Resistance vol.7, pp.5, 2010, https://doi.org/10.5812/jjm.10019
  10. Detection ofSalmonella entericain Magellanic penguins (Spheniscus magellanicus) of Chilean Patagonia: evidences of inter-species transmission vol.143, pp.6, 2010, https://doi.org/10.1017/s0950268814002052
  11. Multiple antibiotic resistances among Shiga toxin producing Escherichia coli O157 in feces of dairy cattle farms in Eastern Cape of South Africa vol.15, pp.None, 2010, https://doi.org/10.1186/s12866-015-0553-y
  12. Serotypes, seasonal trends, and antibiotic resistance of non-typhoidal Salmonella from human patients in Guangdong Province, China, 2009–2012 vol.15, pp.None, 2010, https://doi.org/10.1186/s12879-015-0784-4
  13. Non-Typhoidal Salmonella in poultry meat and diarrhoeic patients: prevalence, antibiogram, virulotyping, molecular detection and sequencing of class I integrons in multidrug resistant strains vol.7, pp.None, 2015, https://doi.org/10.1186/s13099-015-0081-1
  14. Epidemiology, Clinical Presentation, Laboratory Diagnosis, Antimicrobial Resistance, and Antimicrobial Management of Invasive Salmonella Infections vol.28, pp.4, 2010, https://doi.org/10.1128/cmr.00002-15
  15. Genotypic diversity, pathogenic potential and the resistance profile of Salmonella Typhimurium strains isolated from humans and food from 1983 to 2013 in Brazil vol.64, pp.11, 2010, https://doi.org/10.1099/jmm.0.000158
  16. Prevalence of tetracycline resistance determinants in broiler isolatedEscherichia coliin Iran vol.57, pp.6, 2010, https://doi.org/10.1080/00071668.2016.1232478
  17. Quinolone Resistance Mechanisms Among Salmonella enterica in Malaysia vol.22, pp.4, 2010, https://doi.org/10.1089/mdr.2015.0158
  18. Virulence‐associated genes, antimicrobial resistance and molecular typing of Salmonella Typhimurium strains isolated from swine from 2000 to 2012 in Brazil vol.120, pp.6, 2010, https://doi.org/10.1111/jam.13110
  19. Intestinal carriage of multidrug-resistant gram-negative bacteria in preterm-infants during hospitalization in neonatal intensive care unit (NICU) vol.110, pp.6, 2010, https://doi.org/10.1080/20477724.2016.1229884
  20. The effects of pig manure application on the spread of tetracycline resistance in bulk and cucumber rhizosphere soils: a greenhouse experiment vol.63, pp.7, 2017, https://doi.org/10.1139/cjm-2016-0767
  21. Characterization of Quinolone Resistance in Salmonella enterica from Farm Animals in China vol.80, pp.10, 2010, https://doi.org/10.4315/0362-028x.jfp-17-068
  22. Phenotypic and Genotypic Antibiotic Resistance of Salmonella from Chicken Carcasses Marketed at Ibague, Colombia vol.19, pp.2, 2010, https://doi.org/10.1590/1806-9061-2016-0405
  23. Prevalence and characterization of Salmonella among humans in Ghana vol.45, pp.1, 2017, https://doi.org/10.1186/s41182-017-0043-z
  24. Data on antibiogram and resistance genes harboured by Salmonella strains and their Pulsed-field gel electrophoresis clusters vol.17, pp.None, 2018, https://doi.org/10.1016/j.dib.2018.01.098
  25. Prediction of Phenotypic Antimicrobial Resistance Profiles From Whole Genome Sequences of Non-typhoidal Salmonella enterica vol.9, pp.None, 2010, https://doi.org/10.3389/fmicb.2018.00592
  26. Antimicrobial Resistance and Molecular Characterization of Salmonella Enterica Serotypes Isolated from Poultry Sources in Brazil vol.21, pp.1, 2010, https://doi.org/10.1590/1806-9061-2018-0827
  27. Prevalence and characterization of multidrug resistance and variant Salmonella genomic island 1 in Salmonella isolates from cattle, poultry and humans in Iran vol.66, pp.6, 2010, https://doi.org/10.1111/zph.12608
  28. Prevalence and Antibiotic Resistance of Salmonella Enteritidis and Salmonella Typhimurium in Ground Beef and Meatball Samples in Samsun, Turkey vol.26, pp.2, 2010, https://doi.org/10.1089/mdr.2018.0481
  29. Molecular Epidemiology of Infectious Zoonotic and Livestock Diseases vol.8, pp.2, 2010, https://doi.org/10.1128/microbiolspec.ame-0011-2019
  30. Antibiogram Profiles and Risk Factors for Multidrug Resistance of Salmonella enterica Recovered from Village Chickens ( Gallus gallus domesticus Linnaeus) and Other Environmental Sources in the Cen vol.9, pp.10, 2020, https://doi.org/10.3390/antibiotics9100701