Isolation and Characterization of Shiga-toxigenic Escherichia coli Isolated from Various Food Samples

Usaal Tahir¹, Mariam Zameer¹, Nureen Zahra^1, , Maham Mazhar², Ayesha Rafique¹, Sobia Alyas¹, Aroosh Shabbir¹, Arifa Mehreen¹, Muhammad Imtiaz Shafiq², Modasrah Mazhar³ and Qaiser Akram⁴

¹Institute of Molecular Biology and Biotechnology, the University of Lahore, Defence Road, Lahore, Pakistan

²Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan

³Department of Eastern Medicine and Surgery, Qarshi University

⁴University of Veterinary and Animal Sciences Narrowal

Cite this paper:
Usaal Tahir, Mariam Zameer, Nureen Zahra, Maham Mazhar, Ayesha Rafique, Sobia Alyas, Aroosh Shabbir, Arifa Mehreen, Muhammad Imtiaz Shafiq, Modasrah Mazhar and Qaiser Akram. Isolation and Characterization of Shiga-toxigenic Escherichia coli Isolated from Various Food Samples. Journal of Food and Nutrition Research. 2022; 10(1):19-25. doi: 10.12691/jfnr-10-1-3

Abstract

Shiga toxigenic Escherichia coli (STEC) is a food-borne pathogen associated with community health concern. Contamination of food by STEC and toxin causes infectious diseases. Escherichia coli isolated from food samples such as yogurt, apples, tomatoes and milk by applying violet-red bile glucose agar, Triphenyl tetrazolium chloride agar (TTC) and eosin-methylene blue agar (EMB) test after pre-enrichment of food samples in buffered peptone water. Escherichia coli isolates were screened for the existence of shiga toxin genes Stx1 and Stx2 through Polymerase Chain Reaction (PCR). Apples (17%), tomatoes (26%) and milk (1%) of the samples examined showed positive results for Stx genes. Yoghurt samples examined in this study had no Stx1 or Stx2 gene. The presence of Shiga toxin that is protein in nature was confirmed by SDS-PAGE in milk sample. The present study has shown that foods from densely populated areas i.e. Harbanspura, Lahore could be sources of STEC.

Keywords:
E. coli food samples shiga toxin STEC SDS-PAGE

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Mohammadi K. and Karim G.H., 2009. Study on the growth and survival of Escherichia coli O157:H7 during the manufacture and storage of Iranian white cheese in brine. Iran. J. Vet. Res., 10: 346.
[2]	Couturier B.L. 2011. Shiga-Toxigenic Escherichia coli Detection in Stool Samples Screened for Viral Gastroenteritis in Alberta, Canada. J. Microbiol., 49: 574-578.
[3]	Lim J.W.Y. and C.J. Hovde, 2019. Plasmid O157. A Brief Overview of Escherichia coli O157:H7 and Its Plasmid O157. J. Microbiol. Biotechnol., 20: 5-14.
[4]	Luna D.K.F.S., 2009. Characterization of virulence factors in Escherichiacoli isolated from diarrheic and healthy calves in Austria shedding various enteropathogenic agents. Vet. Med., 54(1): 1-11.
[5]	Tsutsuki H., K. Ogura, J. Moss and K. Yahiro, 2020. Host response to the subtilase cytotoxin produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli. Microbiol. Immunol., 64(10): 657-665.
[6]	Collis R.M., P.J. Biggs, A.C. Midwinter, A.S. Browne, D.A. Wilkinson, H. Irshad and A.L. Cookson, 2020. Genomic epidemiology and carbon metabolism of Escherichia coli serogroup O145 reflect contrasting phylogenies. PloS. one., 15(6): 0235066.
[7]	Tumer D.B., 2015. Do the A Subunits Contribute to the Differences in the Toxicity of Shiga Toxin 1 and Shiga Toxin 2. Toxin (basel)., 7: 1468.
[8]	Tanabe R.H.S., M.A. Vieira and N.A.B. Mariano, 2019. Identification and characterization of atypical enteropathogenic and Shiga toxin-producing Escherichia coli isolated from ground beef and poultry breast purchased in Botucatu, Brazil. Braz. J. Microbiol., 50: 1099-1103.
[9]	Liu D., 2009. Introduction. Molecular Detection of Foodborne Pathogens. 1-3.
[10]	Shahzad K.M., 2013. Isolation and molecular characterization of shiga toxin producing E. Coli O157. J. Ani. Plant. Sci., 23(6): 1618-1621.
[11]	Dunn J.R. 2003. Background Shiga-Toxigenic Escherichia coli (STEC).The epidermiology of Shiga-toxigenic Escherichia coli o157:h7 in louisiana dairy cattle, beef cattle, and white-tailed deer. 1.
[12]	Krishnaiah C.B., 2010. Detection of Escherichia coli O157:H7 prevalence in foods of animal origin by cultural methods and PCR technique. Vet. Wrld., 3: 14-15.
[13]	Ames L. 2009. Clinical signs. Enterohemorrhagic Escherichia coli Infections. 3.
[14]	Laura M.S., 2012. E. coli Alpha Hemolysin and Properties. Biochemistry, InTech., 108-110.
[15]	Menge C., 2020. The Role of Escherichia coli Shiga Toxins in STEC Colonization of Cattle. Toxins., 12(9): 607.
[16]	Awofisayo-Okuyelu A, J. Brainard, I. Hall and N. McCarthy, 2019. Incubation period of Shiga toxin–producing Escherichia coli. Epidemiol. rev., 41(1): 121-129.
[17]	Harrison L.M., D.W. Lacher, M.K. Mammel and S.R. Leonard, 2020. Comparative Transcriptomics of Shiga Toxin-Producing and Commensal Escherichia coli and Cytokine Responses in Colonic Epithelial Cell Culture Infections. Fron. Cell. Inf. Microbiol., 10.
[18]	Sambrook J., Amp and D.W. Russell, 2001. Molecular cloning:a laboratory manual (3 volume set)(Vol. 999): Cold spring harbor laboratory press Cold spring Harbor, New York.
[19]	World Health Organization. (2019). Shiga Toxin-producing Escherichia Coli (STEC) and Food: Attribution Characterization and Monitoring (Vol. 19). World Health Organization.
[20]	Abd El, S.S.H. Gany, G.G.F. Mahmoud, S.O. Mousa and R.A. Ibrahem, 2020. Characterization of verotoxigenic E. coli and enteropathogenic E. coli isolated from infants with diarrhea in combination with antimicrobial resistance pattern in Minia. Egypt. J. Adv. Biomed. Pharm. Sci., 3(2): 101-109.
[21]	Karunasagar H.S., 2008. Detection of Shiga-toxigenic Escherichia coli (STEC) in fresh seafood and meat marketed in Mangalore, India by PCR. J. App. Microbiol., 335.
[22]	Bai, X., Fu, S., Zhang, J., Fan, R., Xu, Y., Sun, H., ... & Xiong, Y. (2018). Identification and pathogenomic analysis of an Escherichia coli strain producing a novel Shiga toxin 2 subtype. Scientific reports, 8(1), 1-11.
[23]	Zhang J., Y. Xu, X. Ling, Y. Zhou, Z. Lin, Z. Huang and B. Kan, 2020. Identification of diarrheagenic Escherichia coli by a new multiplex PCR assay and capillary electrophoresis. Mol. Cell. Prob., 49: 101477.
[24]	Heuvelink A.E. 2003. Chapter 16 Review of media for the isolation of diarrhoeagenic Escherichia coli. Prog. Ind. Microbiol., 37: 229-247.
[25]	Shinde, D. B., Singhvi, S., Koratkar, S. S., & Saroj, S. D. (2020). Isolation and characterization of Escherichia coli serotype O157: H7 and other verotoxin-producing E. coli in healthy Indian cattle. Veterinary World, 13(10), 2269.
[26]	Souza M.R.S.M., G. Klassen, F.D. Toni, L.U. Rigo, C. Henkes, C.P. Pigatto, C.B. Dalagassa and C. MTFadel-Picheth, 2010. Biochemical properties, enterohaemolysin production and plasmid carriage of Shiga toxin-producing Escherichia coli strains. Mem. Inst. Oswaldo. Cruz., Rio. De. Janeiro., 105(3): 318-321.
[27]	Brandal, L. T., Wester, A. L., Lange, H., Løbersli, I., Lindstedt, B. A., Vold, L., & Kapperud, G. (2015). Shiga toxin-producing Escherichia coli infections in Norway, 1992-2012: characterization of isolates and identification of risk factors for haemolytic uremic syndrome. BMC infectious diseases, 15(1), 1-10.
[28]	Singh, P., Liu, Y., Bosilevac, J. M., & Mustapha, A. (2019). Detection of Shiga toxin-producing Escherichia coli, stx1, stx2 and Salmonella by two high resolution melt curve multiplex real-time PCR. Food Control, 96, 251-259.
[29]	Reischl U., M.T. Youssef, J. Kilwinski, et al., 2002. Real-time fluorescence PCR assays for detection and characterization of Shiga toxin, intimin, and enterohemolysin genes from Shiga toxin--producing Escherichia coli. J. Clin. Microbiol., 40: 2555-65.
[30]	Pulz M., A. Matussek, M. Monazahian, et al., 2003. Comparison of a Shiga toxin enzyme-linked immunosorbent assay and two types of PCR for detection of Shiga toxin-producing Escherichia coli in human stool specimens. J. Clin. Microbiol., 41: 4671-5.
[31]	El Sayed Z.M. and H. El-Adrosy, 2007. Diagnosis of Shiga toxin producing Escherichia coli infection, contribution of genetic amplification technique. Microbes. Infect., 9: 200-3.
[32]	Ombarak, R. A., Hinenoya, A., Awasthi, S. P., Iguchi, A., Shima, A., Elbagory, A. R. M., & Yamasaki, S. (2016). Prevalence and pathogenic potential of Escherichia coli isolates from raw milk and raw milk cheese in Egypt. International Journal of Food Microbiology, 221, 69-76.
[33]	Okechukwu, E. C., Amuta, E. U., Gberikon, G. M., Chima, N., Yakubu, B., Igwe, J. C., & Njoku, M. (2020). Molecular Identification of Virulence Genes of Escherichia coli Isolated from Cow Milk and Its Products in Abuja, Nigeria. Microbiology Research Journal International, 11-18.
[34]	Martin A. and L. Beutin, 2011. Characteristics of shiga toxin producing Escherichia coli from meat and milk products of different origins and association with food producing animals as main contamination sources. Int. J. Food Microbiol., 146(1): 99-104.
[35]	Brusa, V., Restovich, V., Galli, L., Teitelbaum, D., Signorini, M., Brasesco, H., ... & Leotta, G. A. (2017). Isolation and characterization of non-O157 Shiga toxin-producing Escherichia coli from beef carcasses, cuts and trimmings of abattoirs in Argentina. PLoS One, 12(8), e0183248.