Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
권호 표지
DOI QR코드

DOI QR Code

Colistin resistance and plasmid-mediated mcr genes in Escherichia coli and Salmonella isolated from pigs, pig carcass and pork in Thailand, Lao PDR and Cambodia border provinces

  • Pungpian, Chanika (Research Unit in Microbial Food Safety and Antimicrobial Resistance, Department of Veterinary Public Health, Faculty of Veterinary Science, Chulalongkorn University) ;
  • Lee, Scarlett (Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University) ;
  • Trongjit, Suthathip (Research Unit in Microbial Food Safety and Antimicrobial Resistance, Department of Veterinary Public Health, Faculty of Veterinary Science, Chulalongkorn University) ;
  • Sinwat, Nuananong (Department of Farm Resources and Production Medicine, Faculty of Veterinary Medicine Kasetsart University) ;
  • Angkititrakul, Sunpetch (Department of Veterinary Public Health, Faculty of Veterinary Medicine, Khon Kaen University) ;
  • Prathan, Rangsiya (Research Unit in Microbial Food Safety and Antimicrobial Resistance, Department of Veterinary Public Health, Faculty of Veterinary Science, Chulalongkorn University) ;
  • Srisanga, Songsak (Research Unit in Microbial Food Safety and Antimicrobial Resistance, Department of Veterinary Public Health, Faculty of Veterinary Science, Chulalongkorn University) ;
  • Chuanchuen, Rungtip (Research Unit in Microbial Food Safety and Antimicrobial Resistance, Department of Veterinary Public Health, Faculty of Veterinary Science, Chulalongkorn University)
  • Received : 2021.04.10
  • Accepted : 2021.07.05
  • Published : 2021.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Colistin and carbapenem-resistant bacteria have emerged and become a serious public health concern, but their epidemiological data is still limited. Objectives: This study examined colistin and carbapenem resistance in Escherichia coli and Salmonella from pigs, pig carcasses, and pork in Thailand, Lao PDR, and Cambodia border provinces. Methods: The phenotypic and genotypic resistance to colistin and meropenem was determined in E. coli and Salmonella obtained from pigs, pig carcasses, and pork (n = 1,619). A conjugative experiment was performed in all isolates carrying the mcr gene (s) (n = 68). The plasmid replicon type was determined in the isolates carrying a conjugative plasmid with mcr by PCR-based replicon typing (n = 7). The genetic relatedness of mcr-positive Salmonella (n = 11) was investigated by multi-locus sequence typing. Results: Colistin resistance was more common in E. coli (8%) than Salmonella (1%). The highest resistance rate was found in E. coli (17.8%) and Salmonella (1.7%) from Cambodia. Colistin-resistance genes, mcr-1, mcr-3, and mcr-5, were identified, of which mcr-1 and mcr-3 were predominant in E. coli (5.8%) and Salmonella (1.7%), respectively. The mcr-5 gene was observed in E. coli from pork in Cambodia. Two colistin-susceptible pig isolates from Thailand carried both mcr-1 and mcr-3. Seven E. coli and Salmonella isolates contained mcr-1 or mcr-3 associated with the IncF and IncI plasmids. The mcr-positive Salmonella from Thailand and Cambodia were categorized into two clusters with 94%-97% similarity. None of these clusters was meropenem resistant. Conclusions: Colistin-resistant E. coli and Salmonella were distributed in pigs, pig carcasses, and pork in the border areas. Undivided-One Health collaboration is needed to address the issue.

Keywords

Acknowledgement

The authors wish to thank Dr. Tanittha Chatsuwan, department of microbiology, Chulalongkorn University, for providing control strains for blaKPC, blaNDM-1, blaOXA, blaIMP, and blaVIM.

References

  1. Peng Z, Li X, Hu Z, Li Z, Lv Y, Lei M, et al. Characteristics of carbapenem-resistant and colistin-resistant Escherichia coli co-producing NDM-1 and MCR-1 from pig farms in China. Microorganisms. 2019;7(11):482. https://doi.org/10.3390/microorganisms7110482
  2. Atterby C, Osbjer K, Tepper V, Rajala E, Hernandez J, Seng S, et al. Carriage of carbapenemase- and extended-spectrum cephalosporinase-producing Escherichia coli and Klebsiella pneumoniae in humans and livestock in rural Cambodia; gender and age differences and detection of blaOXA-48 in humans. Zoonoses Public Health. 2019;66(6):603-617. https://doi.org/10.1111/zph.12612
  3. Long H, Feng Y, Ma K, Liu L, McNally A, Zong Z. The co-transfer of plasmid-borne colistin-resistant genes mcr-1 and mcr-3.5, the carbapenemase gene blaNDM-5 and the 16S methylase gene rmtB from Escherichia coli. Sci Rep. 2019;9(1):696. https://doi.org/10.1038/s41598-018-37125-1
  4. Liu L, Feng Y, Zhang X, McNally A, Zong Z. New variant of mcr-3 in an extensively drug-resistant Escherichia coli clinical isolate carrying mcr-1 and blaNDM-5. Antimicrob Agents Chemother. 2017;61(12):e01757-17.
  5. Wang Z, Fu Y, Schwarz S, Yin W, Walsh TR, Zhou Y, et al. Genetic environment of colistin resistance genes mcr-1 and mcr-3 in Escherichia coli from one pig farm in China. Vet Microbiol. 2019;230:56-61. https://doi.org/10.1016/j.vetmic.2019.01.011
  6. Dominguez JE, Redondo LM, Figueroa Espinosa RA, Cejas D, Gutkind GO, Chacana PA, et al. Simultaneous carriage of mcr-1 and other antimicrobial resistance determinants in Escherichia coli from poultry. Front Microbiol. 2018;9:1679. https://doi.org/10.3389/fmicb.2018.01679
  7. Litrup E, Kiil K, Hammerum AM, Roer L, Nielsen EM, Torpdahl M. Plasmid-borne colistin resistance gene mcr-3 in Salmonella isolates from human infections, Denmark, 2009-17. Euro Surveill. 2017;22(31):30587.
  8. Liu YY, Wang Y, Walsh TR, Yi LX, Zhang R, Spencer J, et al. Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China: a microbiological and molecular biological study. Lancet Infect Dis. 2016;16(2):161-168. https://doi.org/10.1016/S1473-3099(15)00424-7
  9. Garcia-Menino I, Diaz-Jimenez D, Garcia V, de Toro M, Flament-Simon SC, Blanco J, et al. Genomic characterization of Prevalent mcr-1, mcr-4, and mcr-5 Escherichia coli within swine enteric colibacillosis in Spain. Front Microbiol. 2019;10:2469. https://doi.org/10.3389/fmicb.2019.02469
  10. European Food Safety AuthorityEuropean Centre for Disease Prevention and Control. The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2017. EFSA J 2019;17(2):e05598.
  11. Olaitan AO, Diene SM, Kempf M, Berrazeg M, Bakour S, Gupta SK, et al. Worldwide emergence of colistin resistance in Klebsiella pneumoniae from healthy humans and patients in Lao PDR, Thailand, Israel, Nigeria and France owing to inactivation of the PhoP/PhoQ regulator mgrB: an epidemiological and molecular study. Int J Antimicrob Agents. 2014;44(6):500-507. https://doi.org/10.1016/j.ijantimicag.2014.07.020
  12. Pungpian C, Sinwat N, Angkititrakul S, Prathan R, Chuanchuen R. Presence and transfer of antimicrobial resistance determinants in Escherichia coli in pigs, pork, and humans in Thailand and Lao PDR border provinces. Microb Drug Resist. 2021;27(4):571-584. https://doi.org/10.1089/mdr.2019.0438
  13. Trongjit S, Angkititrakul S, Tuttle RE, Poungseree J, Padungtod P, Chuanchuen R. Prevalence and antimicrobial resistance in Salmonella enterica isolated from broiler chickens, pigs and meat products in Thailand-Cambodia border provinces. Microbiol Immunol. 2017;61(1):23-33. https://doi.org/10.1111/1348-0421.12462
  14. Sinwat N, Angkittitrakul S, Coulson KF, Pilapil FM, Meunsene D, Chuanchuen R. High prevalence and molecular characteristics of multidrug-resistant Salmonella in pigs, pork and humans in Thailand and Laos provinces. J Med Microbiol. 2016;65(10):1182-1193. https://doi.org/10.1099/jmm.0.000339
  15. Trongjit S, Angkittitrakul S, Chuanchuen R. Occurrence and molecular characteristics of antimicrobial resistance of Escherichia coli from broilers, pigs and meat products in Thailand and Cambodia provinces. Microbiol Immunol. 2016;60(9):575-585. https://doi.org/10.1111/1348-0421.12407
  16. FAO. Monitoring and Surveillance of Antimicrobial Resistance in Bacteria from Healthy Food Animals Intended for Consumption. Bangkok: Food and Agriculture Organization of the United Nations; 2019, 9-26.
  17. U.S. Food and Drug Administration. Enumeration of Escherichia coli and the coliform bacteria [Internet]. Silver Spring: U.S. Food and Drug Administration; https://www.fda.gov/food/laboratory-methods-food/bam-chapter-4-enumeration-escherichia-coli-and-coliform-bacteria. Updated 2017.
  18. ISO. Microbiology of Food and Animal Feeding Stuffs - Horizontal Method for the Detection of Salmonella Spp: ISO6579-Fourth Edition. Geneva: International Organization for Standardization; 2002.
  19. Popoff M, LeMinor L. Antigenic Formulas of the Salmonella Serovars. Paris: Institut Pasteur; 1992.
  20. CLSI. Performance Standards for Antimicrobial Disk and Dilution Susceptibility Test for Bacteria Isolated from Animals 3rd VET01-S3. Wayne: Clinical and Laboratory Standards Institute; 2015.
  21. Dafopoulou K, Zarkotou O, Dimitroulia E, Hadjichristodoulou C, Gennimata V, Pournaras S, et al. Comparative evaluation of colistin susceptibility testing methods among carbapenem-nonsusceptible Klebsiella pneumoniae and Acinetobacter baumannii clinical isolates. Antimicrob Agents Chemother. 2015;59(8):4625-4630. https://doi.org/10.1128/AAC.00868-15
  22. EUCAST. Breakpoint Tables for Interpretation of MICs and Zone Diameters, Version 7.1, Valid from 2017-03-10. Basel: European Committee on Antimicrobial Susceptibility Testing; 2017.
  23. Levesque C, Piche L, Larose C, Roy PH. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob Agents Chemother. 1995;39(1):185-191. https://doi.org/10.1128/AAC.39.1.185
  24. Rebelo AR, Bortolaia V, Kjeldgaard JS, Pedersen SK, Leekitcharoenphon P, Hansen IM, et al. Multiplex PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes. Euro Surveill. 2018;23(6):
  25. Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis. 2011;70(1):119-123. https://doi.org/10.1016/j.diagmicrobio.2010.12.002
  26. Khemtong S, Chuanchuen R. Class 1 integrons and Salmonella genomic island 1 among Salmonella enterica isolated from poultry and swine. Microb Drug Resist. 2008;14(1):65-70. https://doi.org/10.1089/mdr.2008.0807
  27. Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. Identification of plasmids by PCR-based replicon typing. J Microbiol Methods. 2005;63(3):219-228. https://doi.org/10.1016/j.mimet.2005.03.018
  28. Sukhnanand S, Alcaine S, Warnick LD, Su WL, Hof J, Craver MP, et al. DNA sequence-based subtyping and evolutionary analysis of selected Salmonella enterica serotypes. J Clin Microbiol. 2005;43(8):3688-3698. https://doi.org/10.1128/JCM.43.8.3688-3698.2005
  29. Burland TG. DNASTAR's Lasergene sequence analysis software. Methods Mol Biol. 2000;132:71-91.
  30. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018;35(6):1547-1549. https://doi.org/10.1093/molbev/msy096
  31. Zhang X, Zhang B, Guo Y, Wang J, Zhao P, Liu J, et al. Colistin resistance prevalence in Escherichia coli from domestic animals in intensive breeding farms of Jiangsu Province. Int J Food Microbiol. 2019;291:87-90. https://doi.org/10.1016/j.ijfoodmicro.2018.11.013
  32. Huang X, Yu L, Chen X, Zhi C, Yao X, Liu Y, et al. High prevalence of colistin resistance and mcr-1 Gene in Escherichia coli isolated from food animals in China. Front Microbiol. 2017;8:562.
  33. Xu F, Zeng X, Hinenoya A, Lin J. MCR-1 confers cross-resistance to bacitracin, a widely used in-feed antibiotic. MSphere. 2018;3(5):e00411-18.
  34. Strom G, Halje M, Karlsson D, Jiwakanon J, Pringle M, Fernstrom LL, et al. Antimicrobial use and antimicrobial susceptibility in Escherichia coli on small- and medium-scale pig farms in north-eastern Thailand. Antimicrob Resist Infect Control. 2017;6:75. https://doi.org/10.1186/s13756-017-0233-9
  35. Langata LM, Maingi JM, Musonye HA, Kiiru J, Nyamache AK. Antimicrobial resistance genes in Salmonella and Escherichia coli isolates from chicken droppings in Nairobi, Kenya. BMC Res Notes. 2019;12(1):22. https://doi.org/10.1186/s13104-019-4068-8
  36. Varga C, Rajic A, McFall ME, Reid-Smith RJ, Deckert AE, Pearl DL, et al. Comparison of antimicrobial resistance in generic Escherichia coli and Salmonella spp. cultured from identical fecal samples in finishing swine. Can J Vet Res. 2008;72(2):181-187.
  37. Touati M, Hadjadj L, Berrazeg M, Baron SA, Rolain JM. Emergence of Escherichia coli harbouring mcr-1 and mcr-3 genes in North West Algerian farmlands. J Glob Antimicrob Resist. 2020;21:132-137. https://doi.org/10.1016/j.jgar.2019.10.001
  38. Mulvey MR, Bharat A, Boyd DA, Irwin RJ, Wylie J. Characterization of a colistin-resistant Salmonella enterica 4,[5],12:i:- harbouring mcr-3.2 on a variant IncHI-2 plasmid identified in Canada. J Med Microbiol. 2018;67(12):1673-1675. https://doi.org/10.1099/jmm.0.000854
  39. Wise MG, Estabrook MA, Sahm DF, Stone GG, Kazmierczak KM. Prevalence of mcr-type genes among colistin-resistant Enterobacteriaceae collected in 2014-2016 as part of the INFORM global surveillance program. PLoS One. 2018;13(4):e0195281. https://doi.org/10.1371/journal.pone.0195281
  40. Yin W, Li H, Shen Y, Liu Z, Wang S, Shen Z, et al. Novel Plasmid-Mediated Colistin Resistance Gene mcr-3 in Escherichia coli. MBio. 2017;8(3):e00543-17.
  41. Liu L, Feng Y, Zhang X, McNally A, Zong Z. New variant of mcr-3 in an extensively drug-resistant Escherichia coli clinical isolate carrying mcr-1 and blaNDM-5. Antimicrob Agents Chemother. 2017;61(12):e01757-17.
  42. Khine NO, Lugsomya K, Kaewgun B, Honhanrob L, Pairojrit P, Jermprasert S, et al. Multidrug resistance and virulence factors of Escherichia coli harboring plasmid-mediated colistin resistance: mcr-1 and mcr-3 genes in contracted pig farms in Thailand. Front Vet Sci. 2020;7:582899. https://doi.org/10.3389/fvets.2020.582899
  43. Jin L, Wang R, Wang X, Wang Q, Zhang Y, Yin Y, et al. Emergence of mcr-1 and carbapenemase genes in hospital sewage water in Beijing, China. J Antimicrob Chemother. 2018;73(1):84-87. https://doi.org/10.1093/jac/dkx355
  44. Poirel L, Kieffer N, Brink A, Coetze J, Jayol A, Nordmann P. Genetic features of MCR-1-producing colistinresistant Escherichia coli isolates in South Africa. Antimicrob Agents Chemother. 2016;60(7):4394-4397. https://doi.org/10.1128/AAC.00444-16