Antimicrobial susceptibility, biofilm formation, and virulence genes among atypical enteropathogenic Escherichia coli stool isolates in Tehran, Iran
Abstract
Background and Objectives: Enteropathogenic Escherichia coli (EPEC) strains are emerging pathogens around the world, particularly among pediatric patients in developing countries, such as Iran. This study aims to examine and compare the characteristics of EPEC isolates from patients, who suffer from diarrhea versus isolates from patients without diarrhea.
Materials and Methods: A total of 734 stool specimens [440 diarrheal (D), and 294 non-diarrheal (ND)] were examined. Thirty-six EPEC isolates (26 D, and 10 ND) were recovered by culture on MacConkey agar, followed by biochemical tests. Using PCR assay, eae+; stx1- and stx2-gene profiles of EPEC isolates were confirmed. The antimicrobial resistance was assessed by disk diffusion assay. Biofilm formation was assessed using a standard semi-quantitative microtiter plate assay. Virulence-associated genes, ehac, espA, fimA, flu, and sslE were detected.
Results: E. coli comprised 14% of all isolates were EPEC isolates that showed the highest sensitivity to imipenem (IPM) (100%) and gentamicin (GEN) (89%). However, susceptibility to ciprofloxacin and cotrimoxazole or trimethoprim\sulfamethoxazole (SXT) was only 28% and 39%, respectively. About 61% of isolates produced Moderate Biofilm (MB), and the frequency of Weak Biofilm (WB) formers (27%) was higher among D and ND isolates, which carried virulence genes more frequently than D isolates.
Conclusion: Preventive measures by public health authorities can thwart the imminent crisis of widespread zoonotic contamination of the food chain in Iran. Our results may help clinicians make optimal therapeutic choices during the treatment of patients with severe EPEC infections, and assist epidemiologists devise policies for effective control of outbreaks.
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24. Mohammadzadeh M, Oloomi M, Bouzari S. Genetic evaluation of Locus of enterocyte effacement pathogenicity island (LEE) in Enteropathogenic Escherichia coli isolates (EPEC). Iran J Microbiol 2013; 5: 345-349.
25. Aflatoonian MR, Alizade H, Jajarmi M, Ghanbarpour R, Bafti MS, Askari A, et al. Phylotyping of antibiotic resistant, shiga toxin-producing and atypical enteropathogenic Escherichia coli strains isolated from ovine and caprine carcasses in Iran. J Biochem Technol 2018; 9: 41-48.
26. Jomehzadeh N, Ahmadi K, Javaherizadeh H, Afzali M. The first evaluation relationship of integron genes and the multidrug-resistance in class A ESBLs genes in enteropathogenic Escherichia coli strains isolated from children with diarrhea in Southwestern Iran. Mol Biol Rep 2021; 48: 307-313.
27. Puvača N, de Llanos Frutos R. Antimicrobial resistance in Escherichia coli strains isolated from humans and pet animals. Antibiotics (Basel) 2021; 10: 69.
28. Tatar S, Rezatofighi SE, Akhoond MR. Prevalence and antibiotic resistance pattern of diarrheagenic Escherichia coli pathotypes among children under 5 Years in Khuzestan, Iran. Avicenna J Clin Microbiol Infect 2023; 10: 43-48.
29. Dawadi P, Khanal S, Prasai Joshi T, Kc S, Tuladhar R, Maharjan BL, et al. Antibiotic resistance, biofilm formation and sub-inhibitory hydrogen peroxide stimulation in uropathogenic Escherichia coli. Microbiol Insights 2022; 15: 11786361221135224.
30. Faridi F, Abbasi S, Shoja S, Bahador N. Antibacterial activity of chitosan and PAβN on MexAB expression in clinical isolates of ciprofloxacin resistant pseudomonas aeruginosa. Jundishapur J Microbiol 2021; 14(5): e115652.
31. Sicard J-F, Le Bihan G, Vogeleer P, Jacques M, Harel J. Interactions of intestinal bacteria with components of the intestinal mucus. Front Cell Infect Microbiol 2017; 7: 387.
32. Van der Woude MW, Henderson IR. Regulation and function of Ag43 (flu). Annu Rev Microbiol 2008; 62: 153-169.
33. Danese PN, Pratt LA, Dove SL, Kolter R. The outer membrane protein, antigen 43, mediates cell‐to‐cell interactions within Escherichia coli biofilms. Mol Microbiol 2000; 37: 424-432.
34. Ramirez K, Huerta R, Oswald E, Garcia-Tovar C, Hernandez JM, Navarro-Garcia F. Role of EspA and intimin in expression of proinflammatory cytokines from enterocytes and lymphocytes by rabbit enteropathogenic Escherichia coli-infected rabbits. Infect Immun 2005; 73: 103-113.
35. Manning SD, Motiwala AS, Springman AC, Qi W, Lacher DW, Ouellette LM, et al. Variation in virulence among clades of Escherichia coli O157: H7 associated with disease outbreaks. Proc Natl Acad Sci U S A 2008; 105: 4868-4873.
2. Mokomane M, Kasvosve I, de Melo E, Pernica JM, Goldfarb DM. The global problem of childhood diarrhoeal diseases: emerging strategies in prevention and management. Ther Adv Infect Dis 2018; 5: 29-43.
3. Hu J, Torres AG. Enteropathogenic Escherichia coli: foe or innocent bystander? Clin Microbiol Infect 2015; 21: 729-734.
4. Ochoa TJ, Barletta F, Contreras C, Mercado E. New insights into the epidemiology of enteropathogenic Escherichia coli infection. Trans R Soc Trop Med Hyg 2008; 102: 852-856.
5. Mora FX, Avilés-Reyes RX, Guerrero-Latorre L, Fernández-Moreira E. Atypical enteropathogenic Escherichia coli (aEPEC) in children under five years old with diarrhea in Quito (Ecuador). Int Microbiol 2016; 19: 157-160.
6. Mahmoudi-Aznaveh A, Bakhshi B, Najar-Peerayeh S. The trend of enteropathogenic Escherichia coli towards atypical multidrug resistant genotypes. J Chemother 2017; 29: 1-7.
7. Cerutti MF, Vieira TR, Zenato KS, Werlang GO, Pissetti C, Cardoso M. Escherichia coli in chicken carcasses in southern Brazil: absence of shigatoxigenic (STEC) and isolation of atypical enteropathogenic (aEPEC). Braz J Poult Sci 2020; 22: 10.1590/1806-9061-2019-1093.
8. Kostakioti M, Hadjifrangiskou M, Hultgren SJ. Bacterial biofilms: development, dispersal, and therapeutic strategies in the dawn of the postantibiotic era. Cold Spring Harb Perspect Med 2013; 3: a010306.
9. Hernandes RT, De la Cruz MA, Yamamoto D, Girón JA, Gomes TA. Dissection of the role of pili and type 2 and 3 secretion systems in adherence and biofilm formation of an atypical enteropathogenic Escherichia coli strain. Infect Immun 2013; 81: 3793-3802.
10. Llor C, Bjerrum L. Antimicrobial resistance: risk associated with antibiotic overuse and initiatives to reduce the problem. Ther Adv Drug Saf 2014; 5: 229-241.
11. Sirous M, Hashemzadeh M, Keshtvarz M, Amin M, Shams N, Dastoorpoor M, et al. Molecular characterization and antimicrobial resistance of enteropathogenic Escherichia coli in Children from Ahvaz, Iran. Jundishapur J Microbiol 2020; 13(7): e100877.
12. Mahon CR, Lehman DC, Manuselis G (2104). Textbook of diagnostic microbiology: 5th ed. https://shop.elsevier.com/books/textbook-of-diagnostic-microbiology/mahon/978-0-323-08989-0
13. Sjöling Å, Sadeghipoorjahromi L, Novak D, Tobias J. Detection of major diarrheagenic bacterial pathogens by multiplex PCR panels. Microbiol Res 2015; 172: 34-40.
14. Abbey TC, Deak E. What's new from the CLSI subcommittee on antimicrobial susceptibility testing M100, 29th Edition. Clin Microbiol Newsl 2019; 41: 203-209.
15. Rezaei A, Hashemi FB, Heshteli RR, Rahmani M, Halimi S. Frequency of Salmonella serotypes among children in Iran: Antimicrobial susceptibility, biofilm formation, and virulence genes. BMC Pediatr 2022; 22: 557.
16. Abreu AG, Bueris V, Porangaba TM, Sircili MP, Navarro-Garcia F, Elias WP. Autotransporter protein-encoding genes of diarrheagenic Escherichia coli are found in both typical and atypical enteropathogenic E. coli strains. Appl Environ Microbiol 2013; 79: 411-414.
17. Geurtsen J, de Been M, Weerdenburg E, Zomer A, McNally A, Poolman J. Genomics and pathotypes of the many faces of Escherichia coli. FEMS Microbiol Rev 2022; 46: fuac031.
18. Kalule JB, Bester LA, Banda DL, Derra FA, Msefula C, Smith AM, et al. Molecular Epidemiology and AMR Perspective of Diarrhoeagenic Escherichia coli in Africa: A Systematic Review and Meta-analysis. J Epidemiol Glob Health 2024; 14: 1381-1396.
19. Bouzari S, Farhang E, Hosseini SM, Alikhani MY. Prevalence and antimicrobial resistance of shiga toxin-producing Escherichia coli and enteropathogenic Escherichia coli isolated from patients with acute diarrhea. Iran J Microbiol 2018; 10: 151-157.
20. Mansouri M, Mobarez AM, Nojoomi F. Comprehensive study of common Enterogenic E. coli in Iran during 2010–2020: A systematic review. Gene Rep 2021; 25: 101316.
21. Kalantar E, Soheyli F, Salimi H, Soltan DMM. Frequency, antimicrobial susceptibility and plasmid profiles of Escherichia coli pathotypes obtained from children with acute diarrhea. Jundishapur J Microbiol 2011; 4: 23-28.
22. Afset JE, Bevanger L, Romundstad P, Bergh K. Association of atypical enteropathogenic Escherichia coli (EPEC) with prolonged diarrhoea. J Med Microbiol 2004; 53: 1137-1144.
23. Snehaa K, Singh T, Dar SA, Haque S, Ramachandran VG, Saha R, et al. Typical and atypical enteropathogenic Escherichia coli in children with acute diarrhoea: changing trend in East Delhi. Biomed J 2021; 44: 471-478.
24. Mohammadzadeh M, Oloomi M, Bouzari S. Genetic evaluation of Locus of enterocyte effacement pathogenicity island (LEE) in Enteropathogenic Escherichia coli isolates (EPEC). Iran J Microbiol 2013; 5: 345-349.
25. Aflatoonian MR, Alizade H, Jajarmi M, Ghanbarpour R, Bafti MS, Askari A, et al. Phylotyping of antibiotic resistant, shiga toxin-producing and atypical enteropathogenic Escherichia coli strains isolated from ovine and caprine carcasses in Iran. J Biochem Technol 2018; 9: 41-48.
26. Jomehzadeh N, Ahmadi K, Javaherizadeh H, Afzali M. The first evaluation relationship of integron genes and the multidrug-resistance in class A ESBLs genes in enteropathogenic Escherichia coli strains isolated from children with diarrhea in Southwestern Iran. Mol Biol Rep 2021; 48: 307-313.
27. Puvača N, de Llanos Frutos R. Antimicrobial resistance in Escherichia coli strains isolated from humans and pet animals. Antibiotics (Basel) 2021; 10: 69.
28. Tatar S, Rezatofighi SE, Akhoond MR. Prevalence and antibiotic resistance pattern of diarrheagenic Escherichia coli pathotypes among children under 5 Years in Khuzestan, Iran. Avicenna J Clin Microbiol Infect 2023; 10: 43-48.
29. Dawadi P, Khanal S, Prasai Joshi T, Kc S, Tuladhar R, Maharjan BL, et al. Antibiotic resistance, biofilm formation and sub-inhibitory hydrogen peroxide stimulation in uropathogenic Escherichia coli. Microbiol Insights 2022; 15: 11786361221135224.
30. Faridi F, Abbasi S, Shoja S, Bahador N. Antibacterial activity of chitosan and PAβN on MexAB expression in clinical isolates of ciprofloxacin resistant pseudomonas aeruginosa. Jundishapur J Microbiol 2021; 14(5): e115652.
31. Sicard J-F, Le Bihan G, Vogeleer P, Jacques M, Harel J. Interactions of intestinal bacteria with components of the intestinal mucus. Front Cell Infect Microbiol 2017; 7: 387.
32. Van der Woude MW, Henderson IR. Regulation and function of Ag43 (flu). Annu Rev Microbiol 2008; 62: 153-169.
33. Danese PN, Pratt LA, Dove SL, Kolter R. The outer membrane protein, antigen 43, mediates cell‐to‐cell interactions within Escherichia coli biofilms. Mol Microbiol 2000; 37: 424-432.
34. Ramirez K, Huerta R, Oswald E, Garcia-Tovar C, Hernandez JM, Navarro-Garcia F. Role of EspA and intimin in expression of proinflammatory cytokines from enterocytes and lymphocytes by rabbit enteropathogenic Escherichia coli-infected rabbits. Infect Immun 2005; 73: 103-113.
35. Manning SD, Motiwala AS, Springman AC, Qi W, Lacher DW, Ouellette LM, et al. Variation in virulence among clades of Escherichia coli O157: H7 associated with disease outbreaks. Proc Natl Acad Sci U S A 2008; 105: 4868-4873.
| Files | ||
| Issue | Vol 17 No 1 (2025) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v17i1.17799 | |
| Keywords | ||
| Enteropathogenic Escherichia coli (EPEC); Antimicrobial resistance; Biofilm virulence genes | ||
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How to Cite
1.
Halimi S, Rezaei A, Mohebi S, Bonakdar Hashemi F. Antimicrobial susceptibility, biofilm formation, and virulence genes among atypical enteropathogenic Escherichia coli stool isolates in Tehran, Iran. Iran J Microbiol. 2025;17(1):32-40.
