Virulence genes and pulsed- field gel electrophoresis profiles of Shiga toxin-producing Escherichia coli isolated from different food samples and patients with acute diarrhea
,
Abstract
Background and Objectives: Escherichia coli O157: H7 is one of the most important causes of hemorrhagic colitis, and hemolytic uremic syndrome. The present study aimed to isolate E. coli O157: H7 from foods and patients with hemorrhagic colitis, and identify Shiga toxin genes, phylogenetic comparison, and antibiotic resistance of the isolates.
Materials and Methods: In total 400 samples, including patients stool and food were taken in Isfahan-Iran province. Phenotypic tests and PCR were performed to identify Shiga toxin-producing E. coli. The isolated strains were compared phylogenetically by PFGE. Agar disk diffusion was performed to identify the antibiotic resistance of the isolates.
Results: Totally, 5 isolates of fecal samples were E. coli O157, but only 2 isolates carried H7 gene. Also, 9 isolates of E. coli O157 were isolated from food samples that 3 isolates were E. coli O157: H7. The isolates carried stx1, stx2, hlyA and eaeA genes. Also, E. coli non-O157: H7 identified from samples that contained stx1, stx2, hlyA genes. The highest susceptibility to imipenem and the highest resistance to ampicillin and ciprofloxacin were observed. There was a similarity of 100% between the E. coli O157: H7 strains isolated from patients and raw milk and minced beef samples.
Conclusion: Serotypes other than the O157 of E. coli are more prevalent in patients and food. The E. coli O157: H7 isolates from patients had 100% genetic similarity with minced meat and cow milk isolates, which indicates cattle are the most important reservoir of this bacterium in Iran.
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4. Caprioli A, Morabito S, Brugère H, Oswald E. Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res 2005; 36: 289-311.
5. Strachan NJ, Fenlon DR, Ogden ID. Modeling the vector pathway and infection of humans in an environmental outbreak of Escherichia coli O157. FEMS Microbiol Lett 2001; 203: 69-73.
6. Kang'ethe EK, Onono JO, McDermott B, Arimi M. Isolation of E. coli 0157: H7 from milk and cattle feces from urban dairy farming and non-dairy farming neighbor households in Dagoretti Division, Nairobi, Kenya: Prevalence and risk factors. East Afr Med J 2007; 84(11 Suppl): S65-75.
7. Ojo OE, Ajuwape AT, Otesile EB, Owoade AA, Oyekunle MA, Adetosoye AI. Potentially zoonotic Shiga toxin-producing Escherichia coli serogroups in the feces and meat of food-producing animals in Ibadan, Nigeria. Int J Food Microbiol 2010; 142: 214-221.
8. Mersha G, Asrat D, Zewde BM, Kyule M. Occurrence of Escherichia coli O157: H7 in feces, skin, and carcasses from sheep and goats in Ethiopia. Lett Appl Microbiol 2010; 50: 71-76.
9. El-Sayed A, Ahmed S, Awad W. Do camels (Camelus dromedarius) play an epidemiological role in the spread of Shiga Toxin producing Escherichia coli (STEC) infection? Trop Anim Health Prod 2008; 40: 469-473.
10. Tuyet DN, Yassibanda S, Nguyen Thi PL, Koyenede MR, Gouali M, Békondi C, et al. Enteropathogenic Escherichia coli O157 in Bangui and N’Goila, Central African Republic: a brief report. Am J Trop Med Hyg 2006; 75: 513-515.
11. Benkerroum N, Bouhlal Y, El-Attar A, Marhaben A. Occurrence of Shiga toxin-producing Escherichia coli O157 in selected dairy and meat products marketed in the city of Rabat, Morocco. J Food Prot 2004; 67: 1234-1237.
12. Bennani M, Badri S, Baibai T, Oubrim N, Hassar M, Cohen N, et al. First detection of Shiga toxin-producing Escherichia coli in shellfish and coastal environments of Morocco. Appl Biochem Biotechnol 2011; 165: 290-299.
13. Chigor VN, Umoh VJ, Smith SI, Igbinosa EO, Okoh AI. Multidrug resistance and plasmid patterns of Escherichia coli O157 and other E. coli isolated from diarrhoeal stools and surface waters from some selected sources in Zaria, Nigeria. Int J Environ Res Public Health 2010; 7: 3831-3841.
14. Feng P, Weagant SD, Grant MA (2020). BAM Chapter 4: Enumeration of Escherichia coli and the coliform bacteria. Food and Drug Administrations. https://www.fda.gov/food/laboratory-methods-food/bam-chapter-4-enumeration-escherichia-coli-and-coliform-bacteria
15. Mora A, Blanco M, Blanco JE, Dahbi G, López C, Justel P, et al. Serotypes, virulence genes and intimin types of Shiga toxin (verocytotoxin)- producing Escherichia coli isolates from minced beef in Lugo (Spain) from 1995 through 2003. BMC Microbiol 2007; 7: 13.
16. Donohue-Rolfe A, Kondova I, Oswald S, Hutto D, Tzipori S. Escherichia coli O157:H7 strains that express Shiga toxin (Stx) 2 alone are more neurotropic for gnotobiotic piglets than are isotypes producing only Stx1 or both Stx1 and Stx2. J Infect Dis 2000; 181: 1825-1829.
17. Siourimè SN, Isidore BOJ, Oumar T, Nestor BIH, Yves T, Nicolas B, et al, Serotyping and antimicrobial drug resistance of Salmonella isolated from lettuce and human diarrhea samples in Burkina Faso. Afr J Infect Dis 2017; 11: 24-30.
18. Bryce A, Hay AD, Lane IF, Thornton HV, Wootton M, Costelloe C. Global prevalence of antibiotic resistance in pediatric urinary tract infections caused by Escherichia coli and association with routine use of antibiotics in primary care: systematic review and meta-analysis. BMJ 2016; 352: i939.
19. Burow E, Simoneit C, Tenhagen B-A, Käsbohrer A. Oral antimicrobials increase antimicrobial resistance in porcine E. coli--a systematic review. Prev Vet Med 2014; 113: 364-375.
20. Davis MA, Hancock DD, Besser TE, Call DR. Evaluation of Pulsed-Field Gel Electrophoresis as a Tool for Determining the Degree of Genetic Relatedness between Strains of Escherichia coli O157:H7. J Clin Microbiol 2003; 41: 1843-1849.
21. Quinn PJ, Carter ME, Markey B, Carter GR (2002). Clinical Veterinary Microbiology, Mosby, Internal Ltd, London, UK, pp. 42-49.
22. US Food and Drug Administration (FDA). Bacteriological Analytical Manual (BAM), 2024. https://www.fda.gov/food/laboratory-methods-food/bacteriological-analytical-manual-bam
23. Blanco M, Blanco JE, Mora A, Rey J, Alonso JM, Hermoso M, et al. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from healthy sheep in Spain. J Clin Microbiol 2003; 41: 1351- 1356.
24. Rey J, Sanchez S, Blanco JE, Hermoso de Mendoza J, Hermoso de Mendoza M, García A, et al. Prevalence, serotypes, and virulence genes of Shiga toxin-producing E. coli isolated from ovine and caprine milk and other dairy products in Spain. Int J Food Microbiol 2006; 107: 212- 217.
25. Herschleb J, Ananiev G, Schwartz DC. Pulsed-field gel electrophoresis. Nat Protoc 2007; 2: 677-684.
26. Clinical and Laboratory Standards Institute (CLSI) (2018). Performance Standards for Antimicrobial Susceptibility Testing. CLSI Approved Standard M100-S15. Clinical and Laboratory Standards Institute, Wayne.
27. Bonyadian M, Barati S, Mahzonie MR. Phenotypic and genotypic characterization of antibiotic-resistant in Escherichia coli isolates from patients with diarrhea. Iran J Microbiol 2019; 11: 220-224.
28. Bonyadian M, Zahraei Salehi T, Mahzounieh MR, Akhavan Taheri F. Virulence genes of verotoxigenic E. coli isolated from raw milk and unpasteurized cheese. J Vet Res 2011; 66: 223-228.
29. Aslantas O, Erdogan S, Cantekin Z, Gulacti I, Evrendilek GA. Isolation and characterization of verocytotoxin-producing Escherichia coli O157 from Turkish cattle. Int J Food Microbiol 2006; 106: 338-342.
30. Sekhar MS, Sharif NM, Rao TS, Metta M. Genotyping of virulent Escherichia coli obtained from poultry and poultry farm workers using enterobacterial repetitive intergenic consensus-polymerase chain reaction. Vet World 2017; 10: 1292-1296.
31. Vanitha HD, Sethulekshmi C, Latha C. An epidemiological investigation on occurrence of enterohemorrhagic Escherichia coli in raw milk. Vet World 2018; 11: 1164-1170.
32. Prendergast DM, Lendrum L, Pearce R, Ball C, McLernon J, O'Grady D, et al. Verocytotoxigenic Escherichia coli O157 in beef and sheep abattoirs in Ireland and characterization of isolates by Pulsed-Field Gel Electrophoresis and Multi-Locus Variable Number of Tandem Repeat Analysis. Int J Food Microbiol 2011; 144: 519-527.
33. Zhang S, Zhu X, Wu O, Zhang J, Xu X, Li H. Prevalence and characterization of Escherichia coli O157 and O157:H7 in retail fresh raw meat in South China. Ann Microbiol 2015; 65: 1993-1999.
34. Cho SH, Lim YS, Park MS, Kim SH, Kang YH. Prevalence of antibiotic resistance in Escherichia coli fecal isolates from healthy persons and patients with diarrhea. Osong Public Health Res Perspect 2011; 2: 41-45.
35. Selim SA, Ahmed SF, Abdel Aziz MH, Zakaria AM, Klena JD, Pangallo D. Prevalence and characterization of Shiga-toxin O157:H7 and Non-O157:H7 enterohemorrhagic Escherichia coli isolated from different sources. Biotechnol Biotechnol Equip 2013; 27: 3834-3842.
36. Ribot EM, Fair MA, Gautam R, Cameron DN, Hunter SB, Swaminathan B, et al. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne Pathog Dis 2006; 3: 59-67.
37. Swaminathan B, Barrett TJ, Hunter SB, Tauxe RV; CDC PulseNet Task Force. PulseNet: the molecular subtyping network for foodborne bacterial disease surveillance, United States. Emerg Infect Dis 2001; 7: 382-389.
38. Gerner-Smidt P, Hise K, Kincaid J, Hunter S, Rolando S, Hyytiä-Trees E, et al. PulseNet USA: a five-year update. Foodborne Pathog Dis 2006; 3: 9-19.
39. Guh A, Phan Q, Nelson R, Purviance K, Milardo E, Kinney S, et al. Outbreak of Escherichia coli O157 associated with Raw Milk, Connecticut, 2008. Clin Infect Dis 2010; 51: 1411-1417.
40. Jaros P, Cookson AL, Campbell DM, Duncan GE, Prattley D, Carter P, et al. Geographic Divergence of Bovine and Human Shiga Toxin-Producing Escherichia coli O157:H7 Genotypes, New Zealand. Emerg Infect Dis 2014; 20: 1980-1989.
41. Shabana II. Escherichia coli pathotypes associated with diarrhea in human and domestic animals. Am J Anim Vet Sci 2014; 9: 155-161.
42. Gutema FD, Rasschaert G, Agga GE, Jufare A, Duguma AB, Abdi RD, et al. Occurrence, Molecular Characteristics, and Antimicrobial Resistance of Escherichia coli O157 in Cattle, Beef, and Humans in Bishoftu Town, Central Ethiopia. Foodborne Pathog Dis 2021; 18: 1-7.
2. Joensen KG, Engsbro ALØ, Lukjancenko O, Kaas RS, Lund O, Westh H, et al. Evaluating next-generation sequencing for direct clinical diagnostics in diarrhoeal disease. Eur J Clin Microbiol Infect Dis 2017; 36: 1325-1338.
3. Howie H, Mukerjee A, Cowden J, Leith J, Reid T. Investigation of an outbreak of Escherichia coli O157 infection caused by environmental exposure at a scout camp. Epidemiol Infect 2003; 131: 1063-1069.
4. Caprioli A, Morabito S, Brugère H, Oswald E. Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res 2005; 36: 289-311.
5. Strachan NJ, Fenlon DR, Ogden ID. Modeling the vector pathway and infection of humans in an environmental outbreak of Escherichia coli O157. FEMS Microbiol Lett 2001; 203: 69-73.
6. Kang'ethe EK, Onono JO, McDermott B, Arimi M. Isolation of E. coli 0157: H7 from milk and cattle feces from urban dairy farming and non-dairy farming neighbor households in Dagoretti Division, Nairobi, Kenya: Prevalence and risk factors. East Afr Med J 2007; 84(11 Suppl): S65-75.
7. Ojo OE, Ajuwape AT, Otesile EB, Owoade AA, Oyekunle MA, Adetosoye AI. Potentially zoonotic Shiga toxin-producing Escherichia coli serogroups in the feces and meat of food-producing animals in Ibadan, Nigeria. Int J Food Microbiol 2010; 142: 214-221.
8. Mersha G, Asrat D, Zewde BM, Kyule M. Occurrence of Escherichia coli O157: H7 in feces, skin, and carcasses from sheep and goats in Ethiopia. Lett Appl Microbiol 2010; 50: 71-76.
9. El-Sayed A, Ahmed S, Awad W. Do camels (Camelus dromedarius) play an epidemiological role in the spread of Shiga Toxin producing Escherichia coli (STEC) infection? Trop Anim Health Prod 2008; 40: 469-473.
10. Tuyet DN, Yassibanda S, Nguyen Thi PL, Koyenede MR, Gouali M, Békondi C, et al. Enteropathogenic Escherichia coli O157 in Bangui and N’Goila, Central African Republic: a brief report. Am J Trop Med Hyg 2006; 75: 513-515.
11. Benkerroum N, Bouhlal Y, El-Attar A, Marhaben A. Occurrence of Shiga toxin-producing Escherichia coli O157 in selected dairy and meat products marketed in the city of Rabat, Morocco. J Food Prot 2004; 67: 1234-1237.
12. Bennani M, Badri S, Baibai T, Oubrim N, Hassar M, Cohen N, et al. First detection of Shiga toxin-producing Escherichia coli in shellfish and coastal environments of Morocco. Appl Biochem Biotechnol 2011; 165: 290-299.
13. Chigor VN, Umoh VJ, Smith SI, Igbinosa EO, Okoh AI. Multidrug resistance and plasmid patterns of Escherichia coli O157 and other E. coli isolated from diarrhoeal stools and surface waters from some selected sources in Zaria, Nigeria. Int J Environ Res Public Health 2010; 7: 3831-3841.
14. Feng P, Weagant SD, Grant MA (2020). BAM Chapter 4: Enumeration of Escherichia coli and the coliform bacteria. Food and Drug Administrations. https://www.fda.gov/food/laboratory-methods-food/bam-chapter-4-enumeration-escherichia-coli-and-coliform-bacteria
15. Mora A, Blanco M, Blanco JE, Dahbi G, López C, Justel P, et al. Serotypes, virulence genes and intimin types of Shiga toxin (verocytotoxin)- producing Escherichia coli isolates from minced beef in Lugo (Spain) from 1995 through 2003. BMC Microbiol 2007; 7: 13.
16. Donohue-Rolfe A, Kondova I, Oswald S, Hutto D, Tzipori S. Escherichia coli O157:H7 strains that express Shiga toxin (Stx) 2 alone are more neurotropic for gnotobiotic piglets than are isotypes producing only Stx1 or both Stx1 and Stx2. J Infect Dis 2000; 181: 1825-1829.
17. Siourimè SN, Isidore BOJ, Oumar T, Nestor BIH, Yves T, Nicolas B, et al, Serotyping and antimicrobial drug resistance of Salmonella isolated from lettuce and human diarrhea samples in Burkina Faso. Afr J Infect Dis 2017; 11: 24-30.
18. Bryce A, Hay AD, Lane IF, Thornton HV, Wootton M, Costelloe C. Global prevalence of antibiotic resistance in pediatric urinary tract infections caused by Escherichia coli and association with routine use of antibiotics in primary care: systematic review and meta-analysis. BMJ 2016; 352: i939.
19. Burow E, Simoneit C, Tenhagen B-A, Käsbohrer A. Oral antimicrobials increase antimicrobial resistance in porcine E. coli--a systematic review. Prev Vet Med 2014; 113: 364-375.
20. Davis MA, Hancock DD, Besser TE, Call DR. Evaluation of Pulsed-Field Gel Electrophoresis as a Tool for Determining the Degree of Genetic Relatedness between Strains of Escherichia coli O157:H7. J Clin Microbiol 2003; 41: 1843-1849.
21. Quinn PJ, Carter ME, Markey B, Carter GR (2002). Clinical Veterinary Microbiology, Mosby, Internal Ltd, London, UK, pp. 42-49.
22. US Food and Drug Administration (FDA). Bacteriological Analytical Manual (BAM), 2024. https://www.fda.gov/food/laboratory-methods-food/bacteriological-analytical-manual-bam
23. Blanco M, Blanco JE, Mora A, Rey J, Alonso JM, Hermoso M, et al. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from healthy sheep in Spain. J Clin Microbiol 2003; 41: 1351- 1356.
24. Rey J, Sanchez S, Blanco JE, Hermoso de Mendoza J, Hermoso de Mendoza M, García A, et al. Prevalence, serotypes, and virulence genes of Shiga toxin-producing E. coli isolated from ovine and caprine milk and other dairy products in Spain. Int J Food Microbiol 2006; 107: 212- 217.
25. Herschleb J, Ananiev G, Schwartz DC. Pulsed-field gel electrophoresis. Nat Protoc 2007; 2: 677-684.
26. Clinical and Laboratory Standards Institute (CLSI) (2018). Performance Standards for Antimicrobial Susceptibility Testing. CLSI Approved Standard M100-S15. Clinical and Laboratory Standards Institute, Wayne.
27. Bonyadian M, Barati S, Mahzonie MR. Phenotypic and genotypic characterization of antibiotic-resistant in Escherichia coli isolates from patients with diarrhea. Iran J Microbiol 2019; 11: 220-224.
28. Bonyadian M, Zahraei Salehi T, Mahzounieh MR, Akhavan Taheri F. Virulence genes of verotoxigenic E. coli isolated from raw milk and unpasteurized cheese. J Vet Res 2011; 66: 223-228.
29. Aslantas O, Erdogan S, Cantekin Z, Gulacti I, Evrendilek GA. Isolation and characterization of verocytotoxin-producing Escherichia coli O157 from Turkish cattle. Int J Food Microbiol 2006; 106: 338-342.
30. Sekhar MS, Sharif NM, Rao TS, Metta M. Genotyping of virulent Escherichia coli obtained from poultry and poultry farm workers using enterobacterial repetitive intergenic consensus-polymerase chain reaction. Vet World 2017; 10: 1292-1296.
31. Vanitha HD, Sethulekshmi C, Latha C. An epidemiological investigation on occurrence of enterohemorrhagic Escherichia coli in raw milk. Vet World 2018; 11: 1164-1170.
32. Prendergast DM, Lendrum L, Pearce R, Ball C, McLernon J, O'Grady D, et al. Verocytotoxigenic Escherichia coli O157 in beef and sheep abattoirs in Ireland and characterization of isolates by Pulsed-Field Gel Electrophoresis and Multi-Locus Variable Number of Tandem Repeat Analysis. Int J Food Microbiol 2011; 144: 519-527.
33. Zhang S, Zhu X, Wu O, Zhang J, Xu X, Li H. Prevalence and characterization of Escherichia coli O157 and O157:H7 in retail fresh raw meat in South China. Ann Microbiol 2015; 65: 1993-1999.
34. Cho SH, Lim YS, Park MS, Kim SH, Kang YH. Prevalence of antibiotic resistance in Escherichia coli fecal isolates from healthy persons and patients with diarrhea. Osong Public Health Res Perspect 2011; 2: 41-45.
35. Selim SA, Ahmed SF, Abdel Aziz MH, Zakaria AM, Klena JD, Pangallo D. Prevalence and characterization of Shiga-toxin O157:H7 and Non-O157:H7 enterohemorrhagic Escherichia coli isolated from different sources. Biotechnol Biotechnol Equip 2013; 27: 3834-3842.
36. Ribot EM, Fair MA, Gautam R, Cameron DN, Hunter SB, Swaminathan B, et al. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne Pathog Dis 2006; 3: 59-67.
37. Swaminathan B, Barrett TJ, Hunter SB, Tauxe RV; CDC PulseNet Task Force. PulseNet: the molecular subtyping network for foodborne bacterial disease surveillance, United States. Emerg Infect Dis 2001; 7: 382-389.
38. Gerner-Smidt P, Hise K, Kincaid J, Hunter S, Rolando S, Hyytiä-Trees E, et al. PulseNet USA: a five-year update. Foodborne Pathog Dis 2006; 3: 9-19.
39. Guh A, Phan Q, Nelson R, Purviance K, Milardo E, Kinney S, et al. Outbreak of Escherichia coli O157 associated with Raw Milk, Connecticut, 2008. Clin Infect Dis 2010; 51: 1411-1417.
40. Jaros P, Cookson AL, Campbell DM, Duncan GE, Prattley D, Carter P, et al. Geographic Divergence of Bovine and Human Shiga Toxin-Producing Escherichia coli O157:H7 Genotypes, New Zealand. Emerg Infect Dis 2014; 20: 1980-1989.
41. Shabana II. Escherichia coli pathotypes associated with diarrhea in human and domestic animals. Am J Anim Vet Sci 2014; 9: 155-161.
42. Gutema FD, Rasschaert G, Agga GE, Jufare A, Duguma AB, Abdi RD, et al. Occurrence, Molecular Characteristics, and Antimicrobial Resistance of Escherichia coli O157 in Cattle, Beef, and Humans in Bishoftu Town, Central Ethiopia. Foodborne Pathog Dis 2021; 18: 1-7.
| Files | ||
| Issue | Vol 16 No 3 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v16i3.15764 | |
| Keywords | ||
| Food; Human; Shiga toxin-producing E. coli; Pulsed-field gel electrophoresis; Antibiotic resistance | ||
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How to Cite
1.
Bonyadian M, Isvand Haidari F, Sami M. Virulence genes and pulsed- field gel electrophoresis profiles of Shiga toxin-producing Escherichia coli isolated from different food samples and patients with acute diarrhea. Iran J Microbiol. 2024;16(3):329-336.
