Molecular investigation of the prevalence and detection of enterohemorrhagic Escherichia coli isolated from traditional cheese in Iran

authors:

avatar Maryam Maleki parvari , avatar Mahnoosh Parsaeimehr , * , avatar Hamid Staji , avatar Ashkan Jebelli Javan

Koomesh: Vol.23, issue 1; 174-179
published online: December 01, 2021
article type: Research Article
received: February 05, 2020
accepted: June 01, 2020

how to cite: Maleki parvari M, Parsaeimehr M, Staji H, Jebelli Javan A. Molecular investigation of the prevalence and detection of enterohemorrhagic Escherichia coli isolated from traditional cheese in Iran. koomesh. 2021;23(1):e153253. 

Abstract

Introduction: Shiga toxin-producing Escherichia coli (STEC) strains are one of the main causes of food poisoning and diseases such as hemorrhagic colitis and hemolytic uremic syndrome.  Food contamination mainly occurs through food of animal origin such as meat and dairy products. The aim of this study was to investigate prevalence and distribution of virulence genes in Escherichia coli isolates from traditional cheese from Semnan city, using multiplex PCR method. Materials and Methods: In this study, a total of 100 traditional cheese samples were screened for E. coli strains through conventional microbiological tests. Then, screening for virulence genes (Stx1, Stx2, eaeA and hlyA genes) was carried out by using PCR method. Results: Fifty-nine Escherichia coli strains were identified among 100 cheese samples. Of these, 55 isolates exhibited for one or more virulence genes (93.2%) which, 44 isolates carrying more than of one gene and 11 isolates were positive for the carrier of one gene. Stx2 showed the highest frequency of virulence gene in E. coli isolates. Conclusion: Therefore, due to the high prevalence of virulence genes in pathogenic E. coli isolates in traditional cheese, it is necessary to use molecular methods to control the microbial quality of these products.  

References

  • 1.

    Fox PF, Guinee TP, Cogan T, McSweeney PL. Fundamentals of cheese Science. New York 2017.

  • 2.

    Parsaeimehr M, Khazaei M, Jebellijavan A, Staji H. Isolation and identification of dominant lactic acid bacteria by sequencing the 16s rRNA from Semnan traditional cheese (Khiki). J Human Environ Health Prom 2019; 5: 15-20.

  • 3.

    Shekarforoush S, Karim G, Razavi Rohani SM, Kiaie M, Rokni N, Abbasvali M. Study on the overview on foodborne bacteria in foodstuffs with animal origin in Iran; Part one: milk and dairy products. J Food Hygiene 2012; 2.

  • 4.

    Bonyadian M, Zahraei Salehi T, Mahzounie MR, Akhavan Taheri F. Virulence genes of verotoxigenic E. coli isolated from raw milk and unpasturized. J Vet Res 2011; 66: 223-228.

  • 5.

    Conedera G, Dalvit P, Martini M, Galiero G, Gramaglia M, Goffredo E. Verocytotoxin-producing escherichia coli O157 in minced beef and dairy products in Italy. Int J Food Microbiol 2004; 96: 67-73.

  • 6.

    Elhadidy M, Mohammed MA. Shiga toxin producing Escherichia coli from raw milk cheese in Egypt: prevalence, molecular characterization and survival to stress conditions. Lett Appl Microbial 2012; 56: 120-127.

  • 7.

    Rey J, Sanchez S, Blanco JE, Hermoso de Mendoza J, Hermoso de Mendoza M, Garcia A, et al. Prevalence, serotypes and virulence genes of Shiga toxin producing Escherichia coli isolated from ovine and caprine milk and other dairy products in Spain. Int J Food Microbiol 2006; 107: 212-217.

  • 8.

    Solomakos N, Govaris A, Angelidis AS, Pournaras S, Rothi A, Kritas SK, Papageorgiou DK. Occurrence, virulence genes and antibiotic resistance of Escherichia coli O157 isolated from raw bovine, caprine and ovine milk in Greece. Food Microbiol 2009; 26: 865-871.

  • 9.

    Farrokh C, Jordan K, Auvray F, Glass K, Oppegaard H, Raynaud S, Cerf O. Review of Shigatoxin producing Escherichia coli (STEC) and their signi fi cance in dairy production. Int J Food Microbiol 2013; 162: 190-212.

  • 10.

    Baylis CL. Raw milk and raw milk cheeses as vehicles for infection by Verocytotoxin producing Escherichia coli. Int J Dairy Technol 2009; 62: 293-307.

  • 11.

    Hamid S, Salimi Bejestani MR, Changizi E, Javaheri Vayeghan A. Distribution of Escherichia coli Shiga toxin encoding genes (stx1, stx2) in Sangesari lambs suffering from diarrhea by Multiplex PCR technique. Koomesh 2015; 17: 84-91. (Persian).

  • 12.

    Osmundson T, Eyre C, Hayden K. An evaluation of NaOH and alternative rapid DNA extraction protocols for DNA barcoding, genotyping, and disease diagnostics from fungal and oomycete samples. Mol Ecol Resour 2013; 13: 66-74.

  • 13.

    Staji H, Rassouli M, Jourablou S. Comparative virulotyping and phylogenomics of Escherichia coli isolates from urine samples of men and women suffering urinary tract infections. Iranian J Basic Med Sci 2019; 22.

  • 14.

    Paton AW, Paton JC. Direct detection and characterization of Shiga toxigenic escherichia coli by multiplex PCR for stx1, stx 2, eae, ehxA, and saa. J Clin Microbiol 2002; 40: 271-274.

  • 15.

    Safarpoor Dehkordi F, Rahimi E, Ghobadi M, Yahaghi E. Shiga-toxigenic Escherichia oli in sheep cheeses. Iran J Infect Dis 2014; 19: 25-29. (Persian).

  • 16.

    Stephan R, Schumacher S, Corti S, Krause G, Danuser J, Beutin L. Prevalence and characteristics of Shiga toxin-producing Escherichia coli in Swiss Raw milk cheeses collected at producer level. J Dairy Sci 2008; 91: 2561-2565.

  • 17.

    Rezaei M, Yahyaei M, Parviz M, Khodaei motlagh M. A Survey of microbial contamination in traditional cheese distributed in Markazi province in 2010. J Health Environ 2014; 7: 115-122. (Persian).

  • 18.

    Vaziri S, Naghshbandi N. Investigating contamination of traditional non-pasteurized cheeses with Enterobacteriaceae in Maragheh. Iran J Med Microbial 2012; 5: 23-28. (Persian).

  • 19.

    Shadan MR. Investigation of Salmonella Spp, E. coli, listeria monocytogenese, and coagulase positive Staphylococcus in traditional cheese of retail shops in Zahedan. J Res Med Sci 2003; 4: 31-39. (Persian).

  • 20.

    Parsaeimehr M, Jebellijavan A, Azizkhani M, Keykhosravi K, Mahdavi A, Khazaei M. Microbiological and chemichal evaluation of traditional Semnan province khiki cheeses. Appl Animal Sci Res J 2015; 15: 57-64. (Persian).

  • 21.

    Mehrabiyan S, Tahmasby H, Momtaz H, Khosravi N, Kaboli Boroujeni H, Najafzadeh V, et al. Multiplex PCR detection of stx1, stx2 and eaeA genes in Escherichia coli isolated from lambs in Chaharmahal va Bakhtiari, Iran. Biol J Mic 2013; 6: 8-11. (Persian).

  • 22.

    Mahdavi S. Study of frequency of eaeA, stx1 and stx2 genes in Escherichia coli isolated from local cheeses in Maragheh city by multiplex PCR. Iran Food Sci Technol Res J 2016; 12: 388-393.

  • 23.

    Sarimehmetoglu B, Aksoy MH, Ayaz ND, Aya, Y, Kuplulu O, Kaplan YZ. Detection of Escherichia coli O157 H7 in ground beef using immunomagnetic separation and multiplex PCR. Food Cont 2009; 20: 357-361.##.