Detection of Vt1 and Vt2 Genes in E. Coli O157:H7 Isolated from Soft Cheese in Basrah, Iraq Using Duplex Pcr
Keywords:
E. coli, duplex PCR, VT genes, BasrahAbstract
During the period extended from November 2010 to March 2011, One hundred fifty soft cheese samples were collected from three different markets in Basrah city. All samples were cultured in selective media to detect the presence of non-sorbitol fermenting colonies (NSF). 94 (62.66%) isolates were found non-sorbitol fermenting E. coli. Latex agglutination test was used to detect serotype O157:H7 in non- sorbitol fermenting isolates for 15 (33.34%) isolates. Duplex PCR were done to all E. coli O157:H7 isolates and the result showed 5/150 (3.34%) were positive to this test. All the E. coli O157:H7 isolates were positive to vt1 gene which was observed in 100% of isolates but none of the isolates were detected to having a vt2 gene.
References
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Nataro, J. P., Kaper, J. B. (1998). Diarrheagenic Escherichia coli published erratum in Clin Microbiol Rev. Clin. Microbiol. Rev. 11: 142-201.
Niazi, A. D. (2000). Statistical Analysis in Medical Research. Republic of Iraq. Al-Nehrien University. P. 148.
Ogden, I. D.; MacRae, M. and Strachan, N. J. C. (2004). Is the prevalence and shedding concentration of E.coli O157:H7 in beef cattle in Scotland seasonal? F.E.M.S. Microbiol. Lett. 233: 297 – 300.
Paneto, B. R.; Schocken-Iturrino, R. P.; Macedo, C.; Santo, E. and Marin, J. M. (2007). Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil. Arq. Bras. Med. Vet. Zootec. 59: 508-512.
Paton, J. C. and Paton, A. W. (1998). Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli infections. Clin. Microbiol. Rev. 11 (3): 450-479.
Pollard, D. R.; Johnson, W. M.; Lior, H.; Tyler, S. D. and Rozee, K. R. (1990). Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction. J. Clinic. Microbiol. 28: 540-545.
Rahimi, E.; Momtaz, H. and Hemmatzadeh, F.(2008). The prevalence of Escherichia coli O157:H7,Listeria monocytogenes and Compylobacter SPP. on bovine carcasses in Isfahan, Iran. Iranian, J.Vet. Res., Shiraz University 4, 9 (25): 365- 369.
Riley, L. W.; Remis, R. S.; Helgerson, S. D.; McGee, H. B.; Wells, J.G. and Davis, B. R.(1983). Hemorrhagic colitis associated with a rare Escherichia coli serotype. New Engl. J. Med. 308:681-685.
Robert, M. D. (2008). Serotypes and virutypes of enteropathogenic and enterohaemorrhagic E.coli strins from stool samples of children with diarrhea in Germany . J. Appl. Microbiol. 10: 5320-325.
Ropnarine, R. R.; Ammons, D.; Rampersad, J. and Adesiyun, A. A. (2007). Occurance and characterization of verocytotoxigenic Escherichia coli(VTEC) strains from dairy farms in Trindad. J. compilation, Zoonoses Public Health Blackwell verlag, Berlin. 54: 78-85.
Sambrook, J.; Fritsch, E. F. and Maniatis. (1989). Molecular cloning 2nd ed. Cold spring Harbor Laboratory Press, N.Y. 1746-1753.
Sidjabat-Tambunan, H. and Bensink, J. C. (1997). Verotoxin-producing Escherichia coli from the faeces of sheep, calves and pigs. Aust. Vet. J. 75: 292–293.
Spano, G.; Goffredo, E.; Beneduce, L.; Tarantino, D.; Dupuy, A. and Massa, S. (2003). Fate of Escherichia coli O157:H7 during the manufacture of mozzarella cheese. 36:73- 76.
Spano, G.; Goffredo, E.; Beneduce, L.; Tarantino, D.; Dupuy, A. and Massa, S. (2003). Fate of Escherichia coli O157:H7 during the manufacture of mozzarella cheese. 36:73- 76.
Vernozy-Rozand, C.; Montet, M. P.; Beradin, M.; Bavai, C. and Beutin, L. (2005). Isolation and characterization of Shiga toxin-producing Escherichia coli strains from raw milk cheeses in France. Lett. Appl. Microbiol. 41: 235-241.
Vivegnis, J.; Lioui, M.; Leclercq, A. B.; Decallonne, L. J. (1999). Detection of Shiga-like toxin producing Escherichia coli from raw milk cheeses produced in Wallonia. Biotechnol. Agron. Soc. Environ. 3: 159-164.
Yoon, J. W.; . Hovde, C. J. (2008). All blood, no stool: enterohemorrhagi Escherichia coli O157:H7 infection. J. Vet. Sci. 9: 219-231.
Aslam, M.; Hogan, J. and Smith, K.L. (2003). Development of a PCR-based assay to detect Shiga toxin-producing Escherichia coli, Listeria monocytogenes, and Salmonella in milk. Food Microbiol. 20: 345-350.
Buchanan, R.L. and M.P. Doyle. (1997). Foodborne Disease Significance of Escherichia coli O157:H7 and other Enterohemorrhagic E. coli. Food Technol. 51: 69-76.
Centers for Disease Control and prevention (2010). Preliminary Food Net data on the incidence of infection with pathogens transmitted commonly through food-10 states, Morb. and Mortal. Wkly. Rep. (59) 14: 418-422.
Chotar, M.; Vidova, B. and Godany, A. ( 2006). Development of specific and rapid detection of bacterial pathogens in dairy products by PCR. Folia Microbiol. 51: 639-646.
Coia, J.; Johnston, Y.; Steers, N. and Hanson, M. (2001). A survey of the prevalence of Escherichia coli O157:H7 in raw meats, raw cow’s milk and raw-milk cheeses in south-east Scotland. International. J. Food Microbiol. 66: 63-69.
Diez-Gonzalez, F. and Russell, J. B. (1997). The ability of Escherichia coli 0157:H7 to decrease its intracellular pH and resist the toxicity of acetic acid. Microbiol. 143: 1175-1180.
Dontorou, C.; Papadopoulou, C. and Filioussis, G. (2003). Isolation of Esherichia coli O157:H7 from foods in Greece. International. J. Food Microbiol. 82: 273–279.
Doyle, P. M. and Beuchat, L. R. (2007). Food Microbiology: Fundamentals and Frontiers. ASM Press, Washington D.C. Pp. 249-269.
Fagan, P.; Hornitzky, M. A.; Bettelheim, K. A. and Djordjevic, S. P. (1999). Detection of Shiga-like toxin (stx1 and stx2), intimin (eaeA) and enterohemorrhagic Escherichia coli (EHEC) hemolysin (EHEC hlyA) genes in animal feces by multiplex PCR. Appli. and Enviro. Microbiol. 65: 868–872.
Fratamico, P. M. and Smith, J. L. (2006). Escherichia coli infections. In: Riemann, H. P. and Cliver, D. O., Foodborne infections and intoxications. 3rd ed., Florida, Academic Press, an imprint of Elsevier. Pp. 205-208.
Fratamico, P. M.; Sackitey, S. K.; Wiedmann, M. and Deng, M. Y.(1995). Detection of Escherichia coli O157:H7 by multiplex PCR. J. Clin. Microbiol. 33: 2188- 2191.
Hassan, S. A. and Elmalt, L. M. (2008). Informally raw milk and kareish cheese investigation on the occurrence of toxigenic Escherichia coli in Qena city,Egypt with emphasis on molecular characterization. Assiut University. Bull.Environ.Res.11 (2): 35-42.
Ihekweazu, C.; Barlow, M.; Christensen, S. H.; Guttridge, B. and Lewis, D. (2006). Outbreak of E. coli O157 infection in the south west of the UK: risks from streams crossing seaside beaches. Euro. Surveill. 11(4): 128–130.
Karch, H. and Bielaszewska, M. (2001). Sorbitol-fermenting Shiga toxin-producing Escherichia coli O157:H(-) strains: epidemiology, phenotypic and molecular characteristics, and microbiological diagnosis. J. Clinic. Microbiol. 39 (6): 2043-9.
Lim, J. Y.; Yoon, J. W. and Hovde, C. J. (2010). A brief overview of Escherichia coli O157:H7 and its plasmid O157. J. of Microbiol. Biotechnol. 20 (1):1-10.
Maher, M.M. and Murphy,P.M. (2000). Microbiological changes that occur during the ripening of two Irish, smear ripened cheese produced from raw milk. Irish J. Food Res. 39:107-121.
Mashood, A. R.; Minga, U. and Machangu ,R. (2006). Current epidemiological status of enterohaemorrhagic Escherichia coli O157:H7 in Africa. J. Chin. Med. 119 (3): 217–222.
McFadden, J. F. (2000). Biochmical tests .for identification of medical bacteria (3ed), Lippincott Williams and wilkins, USA Microbiol. Rev. 2: 15-38.
Meng, J.; Doyle, M. P.; Zhao, T. and Zhao, S. (2007). Enterohemorrhagic Escherichia coli. in Food Microbiology Fundamentals and Frontiers, 3rd ed. Doyle, M. P. and Beuchat, L. R. ASM Press, Washington D.C. Pp. 249-269.
Muehlherr, J. E.; Zweifel, C.; Corti, S.; Blanco, J. E. and Stephan, R. (2003). Microbiological quality of raw goat’s and ewe’s bulk-tank milk in Switzerland. J. Dairy sci. 86: 3849-3856.
Murinda, S. E.; Nguyen, L. T.; Ivey, S. J.; Gillespie, B. E.; Almeida, R. A.; Draughon, F. A. and Oliver, S. P. (2002). Prevalence and molecular characterization of Escherichia coli O157:H7 in bulk tank milk and fecal samples from cull cows: a 12-month survey of dairy farms in east Tennessee. J. of Food Protec. 65: 752–759.
Nataro, J. P., Kaper, J. B. (1998). Diarrheagenic Escherichia coli published erratum in Clin Microbiol Rev. Clin. Microbiol. Rev. 11: 142-201.
Niazi, A. D. (2000). Statistical Analysis in Medical Research. Republic of Iraq. Al-Nehrien University. P. 148.
Ogden, I. D.; MacRae, M. and Strachan, N. J. C. (2004). Is the prevalence and shedding concentration of E.coli O157:H7 in beef cattle in Scotland seasonal? F.E.M.S. Microbiol. Lett. 233: 297 – 300.
Paneto, B. R.; Schocken-Iturrino, R. P.; Macedo, C.; Santo, E. and Marin, J. M. (2007). Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil. Arq. Bras. Med. Vet. Zootec. 59: 508-512.
Paton, J. C. and Paton, A. W. (1998). Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli infections. Clin. Microbiol. Rev. 11 (3): 450-479.
Pollard, D. R.; Johnson, W. M.; Lior, H.; Tyler, S. D. and Rozee, K. R. (1990). Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction. J. Clinic. Microbiol. 28: 540-545.
Rahimi, E.; Momtaz, H. and Hemmatzadeh, F.(2008). The prevalence of Escherichia coli O157:H7,Listeria monocytogenes and Compylobacter SPP. on bovine carcasses in Isfahan, Iran. Iranian, J.Vet. Res., Shiraz University 4, 9 (25): 365- 369.
Riley, L. W.; Remis, R. S.; Helgerson, S. D.; McGee, H. B.; Wells, J.G. and Davis, B. R.(1983). Hemorrhagic colitis associated with a rare Escherichia coli serotype. New Engl. J. Med. 308:681-685.
Robert, M. D. (2008). Serotypes and virutypes of enteropathogenic and enterohaemorrhagic E.coli strins from stool samples of children with diarrhea in Germany . J. Appl. Microbiol. 10: 5320-325.
Ropnarine, R. R.; Ammons, D.; Rampersad, J. and Adesiyun, A. A. (2007). Occurance and characterization of verocytotoxigenic Escherichia coli(VTEC) strains from dairy farms in Trindad. J. compilation, Zoonoses Public Health Blackwell verlag, Berlin. 54: 78-85.
Sambrook, J.; Fritsch, E. F. and Maniatis. (1989). Molecular cloning 2nd ed. Cold spring Harbor Laboratory Press, N.Y. 1746-1753.
Sidjabat-Tambunan, H. and Bensink, J. C. (1997). Verotoxin-producing Escherichia coli from the faeces of sheep, calves and pigs. Aust. Vet. J. 75: 292–293.
Spano, G.; Goffredo, E.; Beneduce, L.; Tarantino, D.; Dupuy, A. and Massa, S. (2003). Fate of Escherichia coli O157:H7 during the manufacture of mozzarella cheese. 36:73- 76.
Spano, G.; Goffredo, E.; Beneduce, L.; Tarantino, D.; Dupuy, A. and Massa, S. (2003). Fate of Escherichia coli O157:H7 during the manufacture of mozzarella cheese. 36:73- 76.
Vernozy-Rozand, C.; Montet, M. P.; Beradin, M.; Bavai, C. and Beutin, L. (2005). Isolation and characterization of Shiga toxin-producing Escherichia coli strains from raw milk cheeses in France. Lett. Appl. Microbiol. 41: 235-241.
Vivegnis, J.; Lioui, M.; Leclercq, A. B.; Decallonne, L. J. (1999). Detection of Shiga-like toxin producing Escherichia coli from raw milk cheeses produced in Wallonia. Biotechnol. Agron. Soc. Environ. 3: 159-164.
Yoon, J. W.; . Hovde, C. J. (2008). All blood, no stool: enterohemorrhagi Escherichia coli O157:H7 infection. J. Vet. Sci. 9: 219-231.
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2013-06-30
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ABBAS, B. A., & KHUDOR, M. H. (2013). Detection of Vt1 and Vt2 Genes in E. Coli O157:H7 Isolated from Soft Cheese in Basrah, Iraq Using Duplex Pcr. Science Journal of University of Zakho, 1(1), 58–64. Retrieved from https://sjuoz.uoz.edu.krd/index.php/sjuoz/article/view/37
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