Analysis of virulence genes and molecular typing of Listeria monocytogenes isolates from human, food, and livestock from 2008 to 2016 in Iran

Abstract

The frequency of Listeria monocytogenes isolates collected from a total of 1150 samples including food (n = 300), livestock (n = 50), and human clinical (n = 800) was evaluated during 2008–2016. Antimicrobial resistance patterns, virulence factors, and molecular characteristics of these isolates were analyzed using disk diffusion method, sequencing, serotyping, and pulsed-field gel electrophoresis (PFGE). The analysis of 44 L. monocytogenes isolates showed that 72.7% (32 of 44) of all the isolates belonged to Serotype 1/2c, and 15.9% (7 of 44) belonged to Serotype 3c. All 44 isolates were resistant to one or more antimicrobial agents with the most frequent resistance to penicillin (75%) and tetracycline (47.7%). Of the 44 L. monocytogenes strains, 100, 69.2, and 62.5% of livestock, human, and food strains were resistant to penicillin, respectively. Using pulsed-field gel electrophoresis (PFGE) technique, the isolates’ genetic diversity was determined, and 28 PFGE patterns with 8 common (CT) and 20 single types (ST) were identified. This study highlights the high prevalence of Serotype 1/2c in clinical and livestock samples, while different serotypes were observed in food samples. The presence of rare serotypes such as 4c, belonging to the Lineage III, as well as 4e and 1/2c which are infrequent in Iran indicates that paying attention to uncommon serotypes, especially 1/2c, during the listeriosis outbreaks is necessary.

This is a preview of subscription content, access via your institution.

Fig. 1

References

  1. Aarestrup, F.M. and H.C. Wegener, The effects of antibiotic usage in food animals on the development of antimicrobial resistance of importance for humans in Campylobacter and Escherichia coli. Microbes and infection, 1999. 1(8): p. 639-644.

    CAS  Article  Google Scholar 

  2. Ariza-Miguel, J., et al., Molecular epidemiology of invasive listeriosis due to Listeria monocytogenes in a Spanish hospital over a nine-year study period, 2006–2014. BioMed research international, 2015. 2015.

  3. Azizoglu, R.O., L. Gorski, and S. Kathariou, Isolation of Listeria monocytogenes from Food and Water: Official and Experimental Protocols. Current protocols in microbiology, 2014. 33(1): p. 9B. 5.1-9B. 5.19.

    Article  Google Scholar 

  4. Bennett, J.E., R. Dolin, and M.J. Blaser, Mandell, douglas, and bennett's principles and practice of infectious diseases: 2-volume set. Vol. 2. 2014: Elsevier Health Sciences.

  5. Buchanan, R., Risk Assessment of Listeria monocytogenes in Ready-to-eat Foods: Interpretative Summary. 2004: Food & Agriculture Org.

  6. Camargo, A.C., J.J. Woodward, and L.A. Nero, The continuous challenge of characterizing the foodborne pathogen Listeria monocytogenes. Foodborne pathogens and disease, 2016. 13(8): 405-416.

    Article  Google Scholar 

  7. Caplan, M.E., et al., Antibiotic susceptibility profiles of Listeria monocytogenes strains isolated from food products and clinical samples. Revista Romana de Medicina de Laborator, 2014. 22(2): 255-261.

    Article  Google Scholar 

  8. Conter, M., et al., Polymorphism of actA gene is not related to in vitro virulence of Listeria monocytogenes. International journal of food microbiology, 2010. 137(1): p. 100-105.

    CAS  Article  Google Scholar 

  9. Deng, M., et al., Molecular epidemiology and mechanisms of tigecycline resistance in clinical isolates of Acinetobacter baumannii from a Chinese university hospital. Antimicrobial agents and chemotherapy, 2014. 58(1): 297-303.

    Article  Google Scholar 

  10. Fox, E.M., N. Leonard, and K. Jordan, Molecular diversity of Listeria monocytogenes isolated from Irish dairy farms. Foodborne pathogens and disease, 2011. 8(5): p. 635-641.

    CAS  Article  Google Scholar 

  11. Fox, E.M., et al., PFGE analysis of Listeria monocytogenes isolates of clinical, animal, food and environmental origin from Ireland. Journal of Medical Microbiology, 2012. 61(4): 540-547.

    CAS  Article  Google Scholar 

  12. Fugett, E.B., et al., Pulsed-field gel electrophoresis (PFGE) analysis of temporally matched Listeria monocytogenes isolates from human clinical cases, foods, ruminant farms, and urban and natural environments reveals source-associated as well as widely distributed PFGE types. Journal of clinical microbiology, 2007. 45(3): 865-873.

    CAS  Article  Google Scholar 

  13. Gasanov, U., D. Hughes, and P.M. Hansbro, Methods for the isolation and identification of Listeria spp. and Listeria monocytogenes: a review. FEMS microbiology reviews, 2005. 29(5): 851-875.

    CAS  Article  Google Scholar 

  14. Graves, L.M. and B. Swaminathan, PulseNet standardized protocol for subtyping Listeria monocytogenes by macrorestriction and pulsed-field gel electrophoresis. International journal of food microbiology, 2001. 65(1): p. 55-62.

    CAS  Article  Google Scholar 

  15. Haidar-Ahmad, N., et al., Genotypic and virulence characteristics of Listeria monocytogenes recovered from food items in Lebanon. The Journal of Infection in Developing Countries, 2016. 10(07): p. 712-717.

    CAS  Article  Google Scholar 

  16. Indrawattana, N., et al., Prevalence of Listeria monocytogenes in raw meats marketed in Bangkok and characterization of the isolates by phenotypic and molecular methods. Journal of health, population, and nutrition, 2011. 29(1): p. 26.

    Article  Google Scholar 

  17. Jalali, M. and D. Abedi, Prevalence of Listeria species in food products in Isfahan, Iran. International journal of food microbiology, 2008. 122(3): p. 336-340.

    CAS  Article  Google Scholar 

  18. Jamali, H., B. Radmehr, and K.L. Thong, Prevalence, characterisation, and antimicrobial resistance of Listeria species and Listeria monocytogenes isolates from raw milk in farm bulk tanks. Food Control, 2013. 34(1): p. 121-125.

    CAS  Article  Google Scholar 

  19. Jamali, H., et al., Prevalence, antimicrobial susceptibility and virulotyping of Listeria species and Listeria monocytogenes isolated from open-air fish markets. BMC microbiology, 2015. 15(1): p. 144.

    Article  Google Scholar 

  20. Jeffers, G.T., et al., Comparative genetic characterization of Listeria monocytogenes isolates from human and animal listeriosis cases. Microbiology, 2001. 147(5): p. 1095-1104.

    CAS  Article  Google Scholar 

  21. Jensen, A.K., et al., Molecular typing and epidemiology of human listeriosis cases, Denmark, 2002–2012. Emerging infectious diseases, 2016. 22(4): p. 625.

  22. Kathariou, S., Listeria monocytogenes virulence and pathogenicity, a food safety perspective. Journal of food protection, 2002. 65(11): p. 1811-1829.

    Article  Google Scholar 

  23. Kaur, S., et al., Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCR. Journal of applied microbiology, 2007. 103(5): p. 1889-1896.

    CAS  Article  Google Scholar 

  24. Laksanalamai, P., et al., Genomic characterization of novel Listeria monocytogenes serotype 4b variant strains. PloS one, 2014. 9(2):e89024.

    Article  Google Scholar 

  25. Liu, D., Identification, subtyping and virulence determination of Listeria monocytogenes, an important foodborne pathogen. Journal of medical microbiology, 2006. 55(6): p. 645-659.

    Article  Google Scholar 

  26. Liu, D., et al., A multiplex PCR for species-and virulence-specific determination of Listeria monocytogenes. Journal of Microbiological Methods, 2007. 71(2): p. 133-140.

    CAS  Article  Google Scholar 

  27. Lotfollahi, L., et al., Prevalence and antimicrobial resistance profiles of Listeria monocytogenes in spontaneous abortions in humans. African Journal of Microbiology Research, 2011. 5(14): p. 1990-1993.

    Google Scholar 

  28. Lyautey, E., et al., Characteristics and frequency of detection of fecal Listeria monocytogenes shed by livestock, wildlife, and humans. Canadian journal of microbiology, 2007. 53(10): p. 1158-1167.

    CAS  Article  Google Scholar 

  29. Mauro, C., et al., Antimicrobial susceptibility of Listeria monocytogenes isolated from food and food-processing environment. Ann Fac Medic Vet di Parma, 2007. 27: p. 157-164.

    Google Scholar 

  30. Monteiro, L.R.L.D, et al., Molecular characterization of Listeria monocytogenes isolated from animal products in a city of Northern Brazil. Ciência Rural, 2013. 43(8): p. 1443-1448.

    Article  Google Scholar 

  31. Morvan, A., et al., Antimicrobial resistance of Listeria monocytogenes strains isolated from humans in France. Antimicrobial agents and chemotherapy, 2010. 54(6): p. 2728-2731.

    CAS  Article  Google Scholar 

  32. Neves, E., et al., Pulsed-field gel electrophoresis (PFGE) analysis of Listeria monocytogenes isolates from different sources and geographical origins and representative of the twelve serovars. Systematic and applied microbiology, 2008. 31(5): p. 387-392.

    CAS  Article  Google Scholar 

  33. Orsi, R.H., H.C. den Bakker, and M. Wiedmann, Listeria monocytogenes lineages: Genomics, evolution, ecology, and phenotypic characteristics. International Journal of Medical Microbiology, 2011. 301(2): p. 79-96.

    CAS  Article  Google Scholar 

  34. Pan, Y., F. Breidt, and S. Kathariou, Competition of Listeria monocytogenes serotype 1/2a and 4b strains in mixed-culture biofilms. Applied and environmental microbiology, 2009. 75(18): p. 5846-5852.

    CAS  Article  Google Scholar 

  35. Park, S., et al., Molecular Characterization of Listeria monocytogenes Based on the PFGE and RAPD in Korea. Advances in Microbiology, 2012. 2(04): p. 605.

    Article  Google Scholar 

  36. Park, S., et al., Detection of Listeria monocytogenes in Foods and Characterization by PFGE. Advances in Microbiology, 2016. 6(04): p. 343.

    CAS  Article  Google Scholar 

  37. Reniere, M.L., A.T. Whiteley, and D.A. Portnoy, An in vivo selection identifies Listeria monocytogenes genes required to sense the intracellular environment and activate virulence factor expression. PLoS pathogens, 2016. 12(7): e1005741.

    Article  Google Scholar 

  38. Shen, J., et al., Molecular subtyping and virulence gene analysis of Listeria monocytogenes isolates from food. Food microbiology, 2013. 35(1): p. 58-64.

    CAS  Article  Google Scholar 

  39. Soni, D.K., et al., Characterization of Listeria monocytogenes isolated from Ganges water, human clinical and milk samples at Varanasi, India. Infection, Genetics and Evolution, 2013. 14: p. 83-91.

    CAS  Article  Google Scholar 

  40. Strydom, A., et al., Subtyping of Listeria monocytogenes isolated from a South African avocado processing facility using PCR-RFLP and PFGE. Food control, 2013. 31(2): p. 274-279.

    CAS  Article  Google Scholar 

  41. Wagner, M. and F. Allerberger, Characterization of Listeria monocytogenes recovered from 41 cases of sporadic listeriosis in Austria by serotyping and pulsed-field gel electrophoresis. FEMS Immunology & Medical Microbiology, 2003. 35(3): p. 227-234.

    CAS  Article  Google Scholar 

  42. Walsh, D., et al., Antibiotic resistance among Listeria, including Listeria monocytogenes, in retail foods. Journal of Applied Microbiology, 2001. 90(4): p. 517-522.

    CAS  Article  Google Scholar 

  43. Zeinali, T., et al., Serogroup identification and Virulence gene characterization of Listeria monocytogenes isolated from chicken carcasses. Iranian Journal of Veterinary Science and Technology, 2016. 7(2): p. 9-19.

    Google Scholar 

Download references

Acknowledgments

The authors have a special thanks to the staff of Urmia Healthcare Center and Ph.D student of Iran University of Medical Sciences Mrs. Narimani.

Funding

This study was supported by Cellular and Molecular Research Centers of Urmia University of Medical Sciences.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Lida Lotfollahi.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Heidarlo, M.N., Lotfollahi, L., Yousefi, S. et al. Analysis of virulence genes and molecular typing of Listeria monocytogenes isolates from human, food, and livestock from 2008 to 2016 in Iran. Trop Anim Health Prod 53, 127 (2021). https://doi.org/10.1007/s11250-021-02569-7

Download citation

Keywords

  • Listeria monocytogenes
  • Pulsed-field gel electrophoresis
  • Human
  • Food
  • Livestock