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ISSN (Print): 2333-1119 ISSN (Online): 2333-1240 Website: http://www.sciepub.com/journal/jfnr Editor-in-chief: Prabhat Kumar Mandal
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Journal of Food and Nutrition Research. 2014, 2(6), 288-293
DOI: 10.12691/jfnr-2-6-4
Open AccessArticle

Isolation of Enterohaemorrhagic Escherichia coli O104 Strains from Raw Meat Products in the North West Province, South Africa

Collins Njie Ateba1, and Boitumelo Itiel Marumo1

1Department of Biological Sciences, School of Environmental and Health Sciences, Faculty of Agriculture, Science and Technology, North West University, Mafikeng Campus, Private Bag X2046, Mmabatho, South Africa

Pub. Date: June 10, 2014
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Cite this paper:
Collins Njie Ateba and Boitumelo Itiel Marumo. Isolation of Enterohaemorrhagic Escherichia coli O104 Strains from Raw Meat Products in the North West Province, South Africa. Journal of Food and Nutrition Research. 2014; 2(6):288-293. doi: 10.12691/jfnr-2-6-4

Abstract

Enterohaemorrhagic E. coli (EHEC) are Shiga toxin-producing E. coli strains that possess unique pathogenic properties and are characterized by certain seropathotypes that are frequently associated with outbreaks or sporadic cases of human infections. Although EHEC strains belonging to the serotype O104:H4 has rarely been associated with human diseases in the past, outbreaks of infections caused by this organism resulted to HUS and bloody diarrhoea in humans in countries that have advanced public health facilities. The aim of this study was to isolate E. coli O104 strains from meat samples obtained from supermarkets and retail shops in the North West Province, South Africa. A further objective was to determine the identities of the isolates using Gram staining, preliminary biochemical tests (oxidase test, Triple Sugar Iron (TSI) test, citrate utilization and Sorbitol fermentation test) and confirmatory (API 20E and PCR analysis) assays. A total of nineteen meat samples were collected from four butcheries and shops in the study area and Sorbitol MacConkey agar was used for selective isolation of bacteria. A total of 304 presumptive colonies were subjected to identification tests for E. coli. Large proportions (65.5% to 93%) of these isolates were oxidase negative; fermented the sugars in the TSI medium; utilized citrate and fermented sorbitol. On the contrary only a small proportion (27.3%) of the isolates produced hydrogen sulphide gas. Based on patterns obtained for the biochemical profiles of the isolates 102 (33.6%) were positively identified as E. coli. All the 304 E. coli isolates were subjected to specific PCR designed to amplify thewzxO104 gene fragment that facilitates identification of E. coli O104 strains and a total of 52 isolates were positively identified. Despite the fact that not all samples were positive for the pathogen, the presence of E. coli O104 strains in some of the meat samples was of great concern. These bacteria cells have the potential to cause severe health effects on consumers if present in undercooked food products.

Keywords:
Enterohaemorrhagic E. coli (EHEC) E. coli O104 strains food contamination beef diarrhoea wzxO104gene

Creative CommonsThis 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]  Harakeh, S., Yassine, H., Gharios, M., Barbour, E., Hajjar, S., El-Fadel, M., Toufeili, I. and Tannous, R., “Isolation, molecular characterization and antimicrobial resistance patterns of Salmonella and Escherichia coli isolates from meat-based fast food in Lebanon”. Sci Total Environ 134. 33-44. 2005.
 
[2]  Nettleman, M.D.,“Shiga toxin: E. coli O104:H4”. 2011.
 
[3]  Selander, R.K., Musser, J.M., Caugant, D.A., Gilmour, M.N., and Whittam, T.S. “Population genetics of pathogenic bacteria”. Microbial Path 3. 1-7. 1987.
 
[4]  Nataro, J.P. and Kaper, J.B., “Diarrhoeagenic Escherichia coli”.Clin Microbiol Rev11. 142-201. 1998.
 
[5]  Karmali, M. A., “Infection by verotoxin-producing Escherichia coli”. Clin Microbiol Rev 2. 15-38. 1989.
 
[6]  Wieler, L. H., Ilieff, A., Herbst, W., Bauer, C., Vieler, E., Bauerfeind, R., et al “Prevalence of enteropathogens in suckling and weaned piglets with diarrhoea in southern Germany”. J Vet Med B48. 151-159. 2001.
 
[7]  Frank, C., Faber, M.S., Askar, M., Bernard, H., Fruth, A., Gilsdorf, A., Hohle, M., Karch, H., Krause, G., Prager, R., Spode, A., Stark, K. and Werber, D., “HUS investigation team. Large and ongoing outbreak of haemolytic uraemic syndrome, Germany, May 2011”. Euro Surveill16(21).19878. 2011.
 
[8]  Bielaszewska, M., Mellmann, A., Zhang, W., Kock, R., Fruth, A., Bauwens, A., Peters, G. and Karch, H., “Characterisation of the Escherichia coli strain associated with an outbreak of haemolytic uraemic syndrome in Germany, 2011: a microbiological study”. Lancet Infect Dis 11. 671-676. 2011.
 
[9]  Page, A.V. and Liles, W.C., “Enterohaemorrhagic Escherichia coli infections and the Haemolytic-Uraemic Syndrome”. Med Clin North Am 97(4). 681-695. 2013.
 
[10]  Karmali, M.A., Mascarenhas,M., Shen, S., Ziebell, K., Johnson, S., Reid-Smith, R., Isaac-Renton, J., Clark, C., Rahn, K. and Kaper, J.B., “Association of genomic O island 122 of Escherichia coli EDL 933 with verocytotoxin-producing Escherichia coli seropathotypes that are linked to epidemic and/or serious disease. J Clin Microbiol 41. 4930-4940. 2003.
 
[11]  ] Franz, E. and van Bruggen, A.H., “Ecology of E. coli O157:H7 and Salmonella enterica in the primary vegetable production chain. Crit Rev Microbiol 34. 143-161. 2008.
 
[12]  Fremaux, B., Prigent-Combaret, C. and Vernozy-Rozand, C., “Long-term survival of Shiga toxin-producing Escherichia coli in cattle effluents and environment: an updated review”. Vet Microbiol 132 1-18. 2008.
 
[13]  Berger, C.N., Sodha, S.V., Shaw, R.K., Griffin, P.M., Pink, D., Hand, P. and Frankel, G., “Fresh fruit and vegetables as vehicles for the transmission of human pathogens”. Environ Microbiol 12. 2385-2397. 2010.
 
[14]  Bugarel, M., Beutin, L., Martin, A., Gill, A. and Fach, P., “Micro-array for the identification of Shiga toxin-producing Escherichia coli (STEC) seropathotypes associated with Haemorrhagic Colitis and Haemolytic Uremic Syndrome in humans”. Int J Food Microbiol 142. 318-329. 2010.
 
[15]  Capriole, A., Morabito, S., Brugereb, H. and Oswald, E., “Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission”. Vet Res 36. 289-311. 2005.
 
[16]  Mohammed, M.A.M., “Molecular characterization of diarrheagenic Escherichia coli isolated from meat products sold at Monsoura city, Egypt”. Food Cont 25. 159-164. 2012.
 
[17]  Rangel, J.M., Sparling, P.H., Crowe, C., Griffin, P.M. and Swerdlow, D.L, “Epidemiology of Escherichia coli O157:H7 outbreaks, United States, 1982-2002”. Emerg Infect Dis 11. 603-609. 2005.
 
[18]  Erickson, M.C. and Doyle, M.P., “Food as a vehicle for transmission of Shiga toxin- producing Escherichia coli”. Food Protect 70. 2426-2449. 2007.
 
[19]  Miko, A., Delannoy, S., Fach, P., Strockbine, N.A., Lindstedt, B.A., Mariani-Kurkdjian, P., Reetz, J. and Beutin, L., “Genotypes and virulence characteristics of Shiga toxin-producing Escherichia coli O104 strains from different origins and sources”. Int J Med Microbiol 303(8). 410-421. 2013
 
[20]  Scharlach, M., Diercke, M., Dreesman, J., Jahn, N., Krieck, M., Beyrer, K., Claußen, K., Pulz, M. and Floride, R., “Epidemiological analysis of a cluster within the outbreak of Shiga toxin-producing Escherichia coli serotype O104:H4 in Northern Germany, 2011”. Int J Hyg Environ Health 216(3). 341-345. 2013.
 
[21]  Soon, J.M., Seaman, P. and Baines, R.N., “Escherichia coliO104:H4 outbreak from sprouted seeds”. Int J Hyg Environ Health 216(3). 346-354. 2013.
 
[22]  Cramer, J.P., “Enterohaemorrhagic Escherichia coli (EHEC): Haemorrhagic Colitis and Haemolytic Uraemic Syndrome (HUS). Emerg Infect Dis 213-227. 2014.
 
[23]  Rhoades, J.R., Duffy, G. and Koutsoumanis, K., “Prevalence and concentration of verocytotoxigenic Escherichia coli, Salmonella enterica and Listeria monocytogenes in the beef production chain: a review”. Food Microbiol 26. 357-376. 2009.
 
[24]  Karmali, M.A., Gannon, V. and Sargeant, J.M., “Verocytotoxin-producing Escherichia coli (VTEC)”. Vet Microbiol 140. 360-370. 2010.
 
[25]  Browning, N.G., Botha, J.R., Sacho, H. and Moore, P.J., “Escherichiacoli O157:H7 haemorrhagic colitis. Report of the first South African case”. South Afr J Surg 28(1). 28-29. 1990.
 
[26]  Müller, E.E., Ehlers, M.M. and Grabow, W.O.K., “The occurrence of E. coli O157:H7 in Southern African water sources intended for direct and indirect human consumption”. Water Res 35. 3085-3088. 2001.
 
[27]  Müller, E.E., Taylor, M.B., Grabow, W.O.K. and Ehlers, M.M., “Isolation and characterisation of Escherichia coli O157:H7 and shiga toxin-converting bacteriophages from strains of human, bovine and porcine origin”. Water Sup 2(3). 29-38. 2002.
 
[28]  Müller, E.E., Grabow, W.O.K. and Ehlers, M,M., “Immunomagnetic separation of Escherichiacoli O157:H7 from environmental and wastewater in South Africa”. Water SA 29(4). 427-432. 2003.
 
[29]  Müller, E.E. and Ehlers, M.M., “Biolog identification of non-sorbitol fermenting bacteria isolated on E. coli O157 selective CT-SMAC agar”. Water SA 31(2). 247-251. 2005.
 
[30]  Ateba, C.N., Mbewe, M. and Bezuidenhout, C.C., “The Prevalence of Escherichia coli O157 strains in cattle, pigs and humans in the North-West Province, South Africa”. SAJS 104. 78. 2008.
 
[31]  Ateba, C.N. and Bezuidenhout, C.C., “Characterization of Escherichia coli O157 strains from humans, cattle and pigs in the North-WestProvince, South Africa”. Int J Food Microbiol, 128. 181-188. 2008.
 
[32]  Momba, M.N.B., Abongo, B.O. and Mwambakana, J.N., “Prevalence of enterohaemorrhagic Escherichia coli O157:H7 in drinking water and its predicted impact on diarrhoeic HIV/AIDS patients in the Amathole District, Eastern Cape Province, South Africa”. Water SA 34 (3). 365-372.2008.
 
[33]  Ateba, C.N. and Mbewe, M.,“Detection of Escherichia coli O157:H7 virulence genes in isolates from beef, pork, water, human and animal species in the North-West Province, South Africa: public health implications”. Res Microbiol162. 240-248. 2011.
 
[34]  Ateba, C.N. and Mbewe, M.,“Determination of the genetic similarities of fingerprints from Escherichia coli O157:H7 isolated from different sources in the North West Province, South Africa using ISR, BOXAIR and REP-PCR analysis”. Microbiol Res168. 438-446. 2013.
 
[35]  European Centre for Disease Prevention and Control and European Food Safety Authority, 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: ECDC. Available from:
 
[36]  http://ecdc.europa.eu/en/publications/Publications/1106_TER_EColi_joint_EFSA.pdf.
 
[37]  Askar, M., Faber, M.S., Frank, C., Bernard, H., Gilsdorf, A., Fruth, A., Prager, R., Hohle, M., Suess, T., Wadl, M., Krause, G., Stark, K. and Werber, D., “Update onthe ongoing outbreak of haemolytic uraemic syndrome due to Shiga toxinproducing Escherichia coli (STEC) serotype O104, Germany, May 2011”. Euro Surveill 16(22). 19883. 2011.
 
[38]  Brzuszkiewicz, E., Thürmer, A., Schuldes, J., Leimbach, A., Liesegang, H., Meyer, F.D., Boelter, J., Petersen, H., Gottschalk, G. and Daniel, R.,“Genome sequence analyses of two isolates from the recent Escherichia coli outbreak in Germany reveal the emergence of a new pathotype: Entero-Aggregative- Haemorrhagic Escherichia coli (EAHEC)”. Arch Microbiol 193. 883-91. 2011.
 
[39]  Frank, C., Werber, D., Cramer, JP et al.“Epidermic profile of Shiga-toxin-producing Escherichia coli O104:H4 outbreak in Germany- preliminary report”. N Engl J Med. 2011.
 
[40]  Kett, D.H., Quartin, A.A., Mangino, J.E., Zervos, M.J., Ramirez, J.A. and IMPACT – HAP study groups. Escherichia coli O104:H4 South – West France. 2011.
 
[41]  Mellmann, A., Harmsen, D., Cummings, C.A., Zentz, E.B., Leopold, S.R., Rico, A., Prior, K., Szczepanowski, R., Ji, Y., Zhang, W., McLaughlin, S.F., Henkhaus, J.K., Leopold, B., Bielaszewska, M., Prager, R., Brzoska, P.M., Moore, R.L., Guenther, S., Rothberg, J.M. and Karch, H.,“Prospective genomic characterization of the German enterohemorrhagic Escherichiacoli O104:H4 outbreak by rapid next generation sequencing technology”. PLoS One 6(7).e22751.2011.
 
[42]  Borgatta, B., Kmet-Lunaˇcekb, N. and Relloc,J., E. coli O104:H4 outbreak and haemolytic uraemic syndrome”. Med Inten36(8). 576-583. 2012.
 
[43]  Goodridge, L.D., LeJeune, J.T. and Beutin, L., “What was the source of the 2011outbreak of Escherichia coli in Germany and France?”.Case Studies Food Safety Authenticity 43-53. 2012.
 
[44]  Cruishank, R., Duguid, J.P., Marmain, B.P. and Swain, R.H., Medical Microbiology, 12th Edition, New York. Longman Group Limited 2: 34. 1975.
 
[45]  Forbes, A.B. and Weissfeld, A.S., Bailey and Scott’s Diagnostic Microbiology, 10th Edition. Mosby, St. Louis, MO.1998.
 
[46]  Kiiyukia, C., Venkateswaran, K., Navarro, I.M., Nakano, H., Kawakami, H. and Hashimoto, H., “Seasonal distribution of Vibrio parahaemolyticus serotypes along the oyster beds in Hiroshima coast” J Fac Appl Biol Sci, Hiroshima Univ. 28. 49-61. 1989.
 
[47]  Tunung, R., Chai, L.C., Usha, M.R., Lee, H.Y., Fatimah, A.B., Farinazleen, A.B. and Son, R., “Characterization of Salmonella enterica isolated from street food and clinical samples in Malaysia”. ASIAN Food J 14. 161-173.2007.
 
[48]  Bugarel, M., Beutin, L., Martin, A., Gill, A. and Fach, P.,“Micro-array for the identification of Shiga toxin-producing Escherichia coli (STEC) seropathotypes associated with Hemorrhagic Colitis and Hemolytic Uremic Syndrome in humans”. Int J Food Microbiol 142. 318-329. 2010.
 
[49]  Sambrook, J., Fritsch, E.F. and Maniatis, T.’ Molecular Cloning, A Laboratory Manual. 2nd Ed. New York: Cold Spring Harbour Laboratory Press. 1989.
 
[50]  Newell, D.G., Koopmans, M., Verhoef, L., Duizer, E., Aidara-Kane, A., Sprong, H., Opsteegh, M., Langelaar, M., Threfall, J., Scheutz,F., der Giessen, J.V. and Kruse, H., “Foodborne diseases - the challenges of 20 years ago still persist while new ones continue toemerge”. IntJ Food Microbiol 139. 3-15. 2010.
 
[51]  Hussein, H.S., “Prevalence and pathogenicity of Shiga toxin-producing Escherichia coli in beef cattle and their products”. J Ani Sci 85. 63-72. 2007.
 
[52]  Karch, H., Tarr, P.I. and Bielaszewska, M., 2005. Enterohaemorrhagic Escherichia coliin human medicine. Int J Med Microbiol 295, 405-418.
 
[53]  Fach, P., Perelle, S., Dilasser, F. and Grout, J., « Comparison between a PCR-ELISA test and the Vero cell assay for detecting Shiga toxin-producing Escherichia coli in dairy products and characterization of virulence traits of the isolated strains”. J Appl Microbiol 90. 809-818. 2001.
 
[54]  Auvray, F., Lecureuil, C., Tache, J., Leclerc, V., Deperrois, V. and Lombard, B., “Detection, isolation and characterization of Shiga toxin-producing Escherichia coli in retailminced beef using PCR-based techniques, immunoassays and colony hybridization”. Lett ApplMicrobiol 45. 646-651. 2007.
 
[55]  Beutin, L., Miko, A., Krause, G., Pries, K., Haby, S., Steege, K. and Albrecht, N., “Identification of human-pathogenic strains of Shiga toxin-producing Escherichiacoli from food by a combination of serotyping and molecular typing of Shiga toxin genes”. Appl Environ Microbiol 73. 4769-4775. 2007.
 
[56]  Madic, J., Lecureuil, C., Dilasser, F., Derzelle, S., Jamet, E., Fach, P. and Auvray, F., «Screening of food raw materials for the presence of Shiga toxin-producing Escherichia coli O91:H21. Lett Appl Microbiol 48. 447-451. 2009.
 
[57]  Fratamico, P.M., Bagi, L.K., Cray, W.C., Narang, N., Yan, X., Medina, M. and Liu, Y., “Detection by multiplex real-time polymerase chain reaction assays and isolation of Shiga toxin-producing Escherichia coli serogroups O26, O45, O103, O111, O121, and O145 in ground beef”. Foodborne Path Dis 8. 601-607. 2011.
 
[58]  Madic, J., Vingadassalon, N., de Garam, C.P., Marault, M., Scheutz, F., Brugere, H., Jamet, E. and Auvray, F., “Detection of Shiga toxin-producing Escherichia coli serotypes O26: H11, O103:H2, O111:H8, O145:H28, and O157:H7 in raw-milk cheeses by using multiplex real-time PCR”. Appl Environ Microbiol 77. 2035-2041. 2011.
 
[59]  Scheutz, F., Teel, L.D., Beutin, L., Piérard, D., Buvens, G., Karch, H., Mellmann, A., Caprioli, A., Tozzoli, R., Morabito, S., Strockbine, N.A., Melton-Celsa, A.R., Sanchez, M., Persson, S. and O'Brien, A.D.,“Multicenter evaluation of a sequence-based protocol for subtyping Shiga toxins and standardizing Stx nomenclature”. J Clin Microbiol50. 2951-63. 2012.
 
[60]  Maruzumi, M., Morita, M., Matsouka, Y., Uekawa, A., Nakamura, T. and Fugi, K., « Mass food poisoning caused by beef offalcontaminated by Escherichia coli O157”. Jap J Infect Dis 58. 397. 2005.
 
[61]  Auvray, F., Dilasser, F., Bibbal, D., Kérourédan, M., Oswald, E. and Brugère H., “French cattle is not a reservoir of the highly virulent enteroaggregative Shiga toxin-producing Escherichia coli of serotype O104:H4”. Vet Microbiol 158. 443-445. 2012.
 
[62]  Elder, R.O., Keen, J.E., Siragusa, G.R., Barkocy-Gallagher, G.A., Koohmaraie, M. and Laegried, W.W., “Correlation of enterohaemorrhagic Escherichia coli O157 prevalence in faeces, hides and carcasses of beef cattle during processing”. Proc Nat Acad Sci 97. 2999-3003. 2000.
 
[63]  Islam, M.S., Mahmud, Z.H., Gope, P.S., Zaman, R.U., Hossain, Z., Islam, M.S., Mondal, D., Sharker, M.A.Y., Islam, K., Jahan, H., Bhuiya, A., Endtz, H.P., Cravioto, A., Curtis, V., Tour, O., and Cairncross, S., “Hygiene intervention reduces contamination of weaning food in Bangladesh”. Trop Med Int Health 18(3). 250-258. 2013.
 
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