Molecular typing of Clostridium perfringens isolated from minced meat

Document Type : Research Articles

Authors

Ferdowsi University of Mashhad

Abstract

The aim of this study was to determine the presence of toxin genes in Clostridium perfringens isolated from 200minced meat samples using molecular typing. For this purpose, after isolation of Clostridium perfringens using conventional culture method and confirmation by specific 16Srd NA gene PCR, alpha (cpa), beta (cpb), beta 2 (cpb2),epsilon (etx), iota (iA) and enterotoxin (cpe) genes were investigated by multiplex PCR.Out of 200 minced meat samples, 25 (12.5%) were determined as contaminated with C. perfringens and m-PCR results demonstrated that, out of these 25 isolates, 18 (81 %) possessed only the cpa gene (type A), while 4 (18 %) carried the cpa and cpb2 toxin genes (type A-cpb2+). Furthermore, the results showed1 isolate (4 %) as type B, 1 isolate (4 %) as type D and 1 isolate (4 %) as type E. This study reports the first cpb2 positive type B of C. perfringens in minced meat. Our results also indicate that C. perfringens type A is the most common type in minced meat.

Keywords

Main Subjects


Atwa, E.I. and EI-Roos, N.A.A.) 2011( SIncidence of Clostridium perfringensin Meat Products at Some Egyptian Governorat. InternationalJournal of Microbiology Research 2, 196-203.
Augustynowicz, E., Gzyl, A. and Slusarczyk, J. (2000) Molecular epidemiology survey of toxinogenicClostridium perfringens strain types by multiplex PCR. Scandinavian Journal of Infectious Diseases 32, 637–641.
Baums, C.G., Schotte, U., Amtsberg, G., Goethe, R. (2004) Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringensisolates. Veterinary Microbiology 100, 11–16.
Bueschel, D.M., Jost, B.H., Billington, S.J., Trinh, H.T., Songer, J.G. (2003) Prevalence of cpb2, encoding beta2 toxin,inClostridium perfringensfield isolates: correlation ofgenotype with phenotype. Veterinary Microbiology 94,121-129.
Cakmak, O., Ormanci, F.S.B., Tayfur, M. and Erol, I. (2006) Presence and contamination level of Clostridiumperfringens in raw frozen ground poultry and poultry burgers. TurkishJournal of Veterinary and AnimalSciences 30,101–105.
Crespo, R., Fisher, D.J., Shivaprasad, H.L., Fernandez-Miyakawa, M.E., Uzal, F.A. (2007) Toxinotypes of Clostridiumperfringensisolated from sick and healthy avian species. Journal of Veterinary DiagnosticInvestigation 19, 329-333.
Daube, G., Simon, P., Limbourg, B., Manteca, C., Mainil, J., and Kaeckenbeeck, A. (1996) Hybridization of 2,659 Clostridium perfringensisolates with geneprobes for seven toxins and for sialidase.American Journal of VeterinaryResearch 57, 496–501.
Eisgruber, H. and Hauner, G. (2001) Minced beef heart associated with a Clostridium perfringens food poisoning in a Munich old people’s home. Journal of Food Safety and Food Quality 52, 63–66.
Erol, M., Goncuoglu, N.D., Ayaz, FS., Ormanci, B., and Hildebrandt, G. (2008) Molecular typing of Clostridium perfringens isolated from turkey meat by multiplex PCR. Letters in Applied Microbiology 47, 31–34.
Fisher, D.J. (2006) Clostridium perfringense B2 toxin: A potencial Accessory toxin in gastrointestinal disease of human and domestic animals, University of Pittsburg, USA.
Garmory, H.S., Chanter, N., French, N.P., Bueschel, D., Songer, J.G. and Titball, R.W. (2000) Occurrence of Clostridiumperfringens b2-toxin amongst animals, determinedusing genotyping and subtyping PCR assays. Epidemiology & Infection 124, 61–67.
Guran, H.S., Vural, A., Erkan, M.E. (2014) The prevalence and molecular typing of Clostridiumperfringens in ground beef and sheep meats. Journal für Verbraucherschutz und Lebensmittelsicherheit 9,121–128.
Hall, H.E. and Angelotti, R. (1965) Clostridium perfringens in Meat and Meat Products. Journal of Applied Microbiology 13.
Hatakka, M. and Halonen, H. (2000) Food-borne and Waterborne Outbreaks in Finland in 1999. National Food Administration Research Notes 7.
Hatheway, C.L. (1990) Toxigenic clostridia. Clinical Microbiology Review 3, 66-98.
Heikinheimo, A., Lindström, M., Korkeala, H. (2004) Enumerationand isolation of cpe-positive Clostridium perfringensspores from feces. Journal of ClinicalMicrobiology 42, 3992-3997.
Hughes, C., Gillespie, I.A. and O’Brien, S.J. (2007) Foodborne transmission of intestinal disease in England and Wales, 1992–2003. Food Control 18, 766–772.
Kadra, B., Gouillou, J.P., Popoff, M. and Bourlioux, P. (1999) Typing of sheep clinical isolates and identification of enterotoxigenic Clostridium perfringens strains by classical methods and by polymerase chain reaction (PCR). FEMS Immunologyand Medical Microbiology 24, 259–266.
Kanakaraj, R., Harris, D.L., Songer, J.G. and Bosworth, B. (1998) Multiplex PCR assay for detection of Clostridium perfringens in feces and intestinal contents of pigs and inswine feed. Veterinary Microbiology 63, 29–38.
Kamber, U., Gokce, H.I., Elmali, M. (2007) Clostridium perfringens and its toxins in minced meat from Kars.Food Additives & Contaminants 24, 673-688.
McClane, B.A. (2001) Clostridium perfringens. In Food Microbiology: Fundamentals and Frontiers ed. Doyle, M.P., Beuchat, L.R. and Montville, T.J. Washington,DC: ASM Press, pp. 351–372.
McDonel, J.L. (1986) Toxins of Clostridium perfringens types A, B, C, D, and E. In Pharmacology of Bacterial Toxins ed. Dorner, F and Drews, H. Oxford: Pergamon Press, pp. 477–517.
Meer, R. and Songer, J.G. (1997) Multiplex polymeraseChain reaction assay for genotyping Clostridium perfringens. American Journal of Veterinary Research 58, 702-705.
Novak, J.S., Juneja, V.K. (2002) Clostridium perfringens: hazards in new generation foods. Innov. Food Science Emerg. Technology 3, 127_132.
Oakley, C. and Wayrack, G. (1953) Routine typing of Clostridium welchii. Journal of Hygiene 51, 102–107.
Razmyar, J., Kalidari, G.A., Tolooe, A., Rad, M. and Movassaghi, A.R. (2014). Genotyping of Clostridium perfringens isolated from healthy and diseased ostriches (Struthiocamelus). Iranian Journal of Microbiology 6, 31-36.
Ridell, J., Bjo¨rkroth, J., Eisgru¨ber, H., Schalch, B., Stolle, A. and Korkeala, H. (1998) Prevalence of the enterotoxin gene and clonality of Clostridium perfringens strains associated with food-poisoning outbreaks. Journal of Food Protection 61, 240–243.
Songer, J.G. (1996) Clostridial enteric diseases of domestic animals. Clinical Microbiology Review 9, 216-234.
Songer, J.G. and Meer, R.M. (1996) Genotyping of Clostridium perfringens by polymerase chain reaction is a useful adjunct to diagnosis of clostridial enteric disease in animals. Anaerobe 2, 197–203.
Sterne, M. and Batty, I. (1975) Criteria for diagnosing clostridial infection. In Pathogenic Clostridia ed. Sterne, M. and Batty, L. London: Butterworths, pp. 79–122.
Torky, A.A.S (2004). Trials for inhibition of some food poisoning microorganisms in meat products, Cairo University, Egypt.
Wen, Q., and McClane, B.A. (2004) Detection of enterotoxicogenic clostridium perfringens Type A isolates in American retail foods. Applied Environmental Microbiology 70, 2685–2691.
Wu, J., Zhang, W., Xie, B., Wu, M., Tong, X., Kalpoe, J., Zhang, D. (2009) Detection and Toxin Typing of Clostridium perfringensin Formalin-Fixed, Paraffin-Embedded Tissue Samples by PCR. Journal of Clinical Microbiology 47, 807-810.
Yoo, H.S., Lee, S.U., Park, K.Y., Park, Y.H. (1997). Moleculartyping and epidemiological survey of prevalence ofClostridium perfringens types by multiplex PCR. Journal of Clinical. Microbiology 35, 228–232.
 
CAPTCHA Image