Shiga toxin (STX) gene analysis and verotoxigenic potentials of Escherichia coli isolated from ‘bobozi’- an indigenous Nigerian ready to eat fermented cassava chip

  • Authors

    • Stephen Enabulele Benson Idahosa University, Benin City
    • Ebakota Daniel Benson Idahosa University
    2016-08-29
    https://doi.org/10.14419/ijbr.v4i2.6543
  • Bobozi, Verotoxins, Escherichia coli, Shiga toxins, Vero cells.
  • Abstract

    Many strains of E. coli have been identified as food borne pathogens inducing serious gastrointestinal diseases and even causing death in humans. Amongst these are those referred to as Shiga or Vero toxin producing E. coli. Bobozi is an Indigenous Nigerian ready to eat snack food made from cassava and is widely consumed in most parts of the country. The objective of this study was to Isolate and Identify E. coli from Bobozi, analyze their Shiga toxin (stx) gene profile and thereafter determine their verotoxin capabilities. A total of 248 samples of Bobozi were purchased from hawkers in the metropolis of Benin City, Edo state, Nigeria. Isolation and Identification was done using standard microbiological methods. Analysis for the presence of Shiga toxin (stx1 and stx2) genes was done using PCR technique and for their verotoxigenic potentials using tissue culture assay on Vero cells. Result of the investigation reveals that of the total 248 samples collected, 107(43.15%) had E. coli isolated from them. Total number of E. coli isolated from the 107 samples was 386.Shiga toxin gene analysis of the isolates reveal that 169(43.78%) had stx1 gene alone, 112 (29.02%) had stx2 gene alone, 3(0.78%) had both stx 1 and stx 2 genes while 102 (26.42%) had none of the two genes. Tissue culture assay on Vero cells indicate that there is a strong relationship between the presence of stx genes and their degree of cytotoxic effect on vero cells as over 80% isolates without the genes exhibited weak cytotoxic effect whereas over 80% of those with the genes exhibited more than strong cytotoxic effect.

  • References

    1. [1] Bennett J. and Bettelheim K. A. (2002). Serotypes of non-O157 verocytotoxigenic Escherichia coli isolated from meat in New Zealand. Comp Immunol Microbiol Infect Dis. 25(2):77-84http://dx.doi.org/10.1016/S0147-9571(01)00024-8.

      [2] Beutin L, Krause G, Zimmermann S, Kaulfuss S, Gleier K. (2004). Characterization ofShiga toxin-producing Escherichia coli strains isolated from human patients in Germany over a 3-year period. J ClinMicrobiol. 42(3):1099-108http://dx.doi.org/10.1128/JCM.42.3.1099-1108.2004.

      [3] Blanco M., Blanco J.E., Mora A., Ray J., Alonso J. M., Hermoso M., Hermoso J., Alonso M.P., Dhabi G., Gonzalez E. A., Bernardez E. I. and Blanco J.(2003). Serotypes, virulence genes and intimin types of Shiga toxin (verotoxin) producing Escherichia coli isolates from healthy sheep in Spain. J. Clin. Microbiol. 42: 645 – 651.http://dx.doi.org/10.1128/JCM.42.2.645-651.2004.

      [4] Egan, M.B., Raats, M.M., Grubb, S.M., Eves, A., Lumbers, M.L., Dean M.S. and Adams, M.R. (2007). A review of food safety and food hygiene training studies in the commercial sector. Food Control, 18: 1180-1190.http://dx.doi.org/10.1016/j.foodcont.2006.08.001.

      [5] Enabulele, S. A. and Uraih, N. (2009). Enterohaemorrhagic Escherichia coli 0157:H7Prevalence in meat and vegetables sold in Benin City, Nigeria. Afri. J. of Microbiol. Res. 3(5): 276-279.

      [6] Enabulele S.A., Eghafona N.O. and Dahiru M.(2015). Molecular Characterisation and Verotoxigenic Potentials of Enterohaemorrhagic Escherichia coli 0157:H7 Isolated from Fermented Fresh Cow Milk (Nunu) sold in Selected Cities in Nigeria. BIU Journal of basic and Applied Sciences. 1(1): 51-62.

      [7] European Centre for Disease Prevention and Control and European Food Safety Authority (ECDC) (2011). Shiga toxin/verotoxin-producing Escherichia coli in humans, food and animals in the EU/EEA, with special reference to the German outbreak strain STEC O104. Stockholm.

      [8] Fey, P. D., Wickert, R. S., Rupp, M. E., Safranek, T.J. and Hinrichs, S.H. (2000). "Prevalence of Non-O157:H7 Shiga Toxin-Producing Escherichia coli in Diarrheal Stool Samples from Nebraska" Public Health Resources. Paper 21. http://digitalcommons.unl.edu/publichealthresources/21.

      [9] Gyles C. L. (2007). Shiga toxin- producing Escherichia coli: An overview. J. Anim. Sci. 85: E45-E62http://dx.doi.org/10.2527/jas.2006-508.

      [10] Holt, J. G, Krevy, H. S, Sneathe, R. H. A, Williams S. T. (1994). Bergey’s Manual of Determinative Bacteriology 9th Edition. Williams and Wilkens Company, Baltimore, USA.

      [11] Hussein H. S. (2007). Prevalence and Pathogenicity of Shiga- toxin producing Escherichia coli in beef cattle and their products. J. Anim. Sci. 85: E63-E72.http://dx.doi.org/10.2527/jas.2006-421.

      [12] Kaplan B. S, Cleary T. G, Obrig T. G. (1990). Recent advances in understanding the pathogenesis of the haemolytic uremic syndrome. Pediatr Nephrol. 4: 276-83.http://dx.doi.org/10.1007/BF00857676.

      [13] Konowalchuk, J., Speirs, J. I., and Stavric, S. (1977). Vero response to a cytotoxin of Escherichia coli. Infect. Immun. 18:775–779.

      [14] Mathusa E. C., Chen Y., Enache E. Hontz L. (2010). Non-O157 Shiga toxin producing Escherichia coli in foods. J. Food Prot. 73(9):1721-36.

      [15] Mensah, P., Yeboah, M. D., Owusu, D. K. and Ablordey, A. (2002). Street foods in ACCRA, Ghana: how safe are they? Bulletin of the World Health Organization 80(7): 546-554.

      [16] Nataro, J. P. and Kaper J. B. (1998). Diarrheagenic Escherichia coli Clin. Microbiol. Rev. 11: 142 – 201

      [17] Oranusi, U. S. and Braide, W. (2012). A study of microbial safety of Ready-to-eat foods vended on highways: Onitsha-Owerri, south east Nigeria. International Research Journal of Microbiology 3(2):006-071.

      [18] Rahn K., Wilson J. B. Mcfadden K. A. Read S. C. Ellis A. G.Renwick S. A., Clarke R. C and Johnson R. P.(1996). Comparison of Vero Cell Assay and PCR as Indicators of the Presence of Verocytotoxigenic Escherichia coli in Bovine and Human Fecal Samples. Appl. Environ. Microbiol. 62(12): 4314 – 4317.

      [19] Sambrook J. and Russell, D. W.(2001). Molecular cloning: A Laboratory Manual vol 2. Cold Spring Habour Lab. Press New York. p126.

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  • How to Cite

    Enabulele, S., & Daniel, E. (2016). Shiga toxin (STX) gene analysis and verotoxigenic potentials of Escherichia coli isolated from ‘bobozi’- an indigenous Nigerian ready to eat fermented cassava chip. International Journal of Biological Research, 4(2), 177-180. https://doi.org/10.14419/ijbr.v4i2.6543

    Received date: 2016-07-29

    Accepted date: 2016-08-24

    Published date: 2016-08-29