Electrophoretic characterization for detection of Clostridium perfringens enterotoxin type ( A ) in meat. | ||||
Assiut Veterinary Medical Journal | ||||
Article 1, Volume 53, Issue 115 - Serial Number 4, October 2007, Page 1-11 PDF (3.36 MB) | ||||
Document Type: Research article | ||||
DOI: 10.21608/avmj.2007.167008 | ||||
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Author | ||||
, A.M.O El-Mahrouk | ||||
Microbial Toxins Research Unit Animal Health Research Institute, Dokki, Giza | ||||
Abstract | ||||
A total of 100 buffalo meat samples were collected from different markets in Giza govemorate. They were subjected to physical and bacteriological examination. Thirty nine percentage of the samples tested were found to be contaminated with Clostridium perfringens type A. The most probable number (MPN/gram) values ranging from 0 to 35. Also, the spore count of Clostridium perfringens type A at 100°C at different time interval was investigated. Electrophoretic analysis and immunobloting for detection of Clostridium perfringens enterotoxin type (A) was carried out for its characterization. The immunogenic band was determined with molecular | ||||
Keywords | ||||
Key words: Electrophoresis Cl. perfringens; enterotoxin; meat | ||||
Full Text | ||||
Assiui Vet. Med. J. Voi. 53 No. 115 October 2007 Microbial Toxins Research Unit _ ELECTROPHORETIC CHARACTERIZATION FOR ENTEROTOXIN TYPE (A) IN MEAT By cond 8 ye ayes plus ISI Cag Se Ppa gtd fl gS) Cine gill Lael pat dans cible Bes) yo aes oo Gagdall pal Gye Ake Mls we ole Coll Ie gai SUMMARY A total of 100 buffalo meat samples were collected from different markets in Key words: Electrophoresis Cl. perfringens, enterotoxin, meat a INTRODUCTION Most species of the genus Clostridium are saprophytes that Clostridium perfringens type A isolates can carry the enterotoxin In vitro toxin production is an important tool not only for The prevalence of the enterotoxin gene in a well-characterized Meat and fish are sensitive to contamination and support growth Clostridium perfringens type A is one of the four most important Cl. perfringens carried in the human and animal intestine, soil, The isolation of Cl. botulinum from foods is generally considered Seven types of CL botulinum (A, B, C, D, E F and G) are a on the basis of antigenic Specificity of their toxins; Pierson ef eee The objective of this study was to investigate the role of several 2-Detection of the ability of of Clostridium perferingens type A to 3-Detection of the of Clostridium perferingens type A in foods of 4-Western Blotting or immunoblotting for identification of MATERIALS and METHODS Samples: A total of 100 buffalo meat samples were collected from Cultural conditions: Processing of each food sample started with a Each FTG enrichment culture showing growth was streaked onto 3
Administration, 1998. A loopful of each culture was stabbed into a tube Determination of MPN of CL. perfringens per gram in meat: A three-lube most probable number (MPN) method was used to Statistical analyses were performed; Koburger, 1975. To evaluate the heat resistance of Cl perfringens meat isolates, Briefly, sporulating cultures of C/ perfringens were prepared by The remainder of each heat-shocked DS culture was then heated determine The number of viable spores present at th ime point per Detection of enterotoxins producing isolates by reversed passive A porlion of the sporulated culture (about 5 m!) was centrifuged Solubilized cells (0.4 gm/ml) were mixed with an equal volume After electrophoresis, the separated proteins were transferred to RESULTS
aus No. of samples
Table 2: Spore count of Cl. perfringens type A after heating at 100°C at
Isolates 0 30 min. 60 min. 90min. | 120 min. I a | 3.4 | 0 0 10 2 2 3.31 22 1S id 3 17 | 4.0 LI 10 0
Western Blot analysis of CPE, Lane A: Molecular weight marker; Lane DISCUSSION Clostridium perfringens isolates are commonly classified into 6 To date, there are at least two explanations for the strong As shown in Table (1) 39.0% of meat samples tested in the In the provisional, 39.0% of meat samples grew Cl. perfringens Little is known about the mechanisms responsible for the specific eZ Assiut Vet. Med. J. Vol. 53 No. 115 October 2007 Raju and Sarker, 2005 showed that Gi) the heat resistance of With the ultimate goal of better controlling Ci. perfringens type The enteropathogenic effects of CPE are primarily mediated Deaths from C perfringens type A food poisoning are not Results from the present study provide an explanation for the REFERENCES Abdel-Rahman, M.; Abd-Allah, W.H. and Abd El-Aziz, S (1996): 8 Barnes, E.M. (1985): Isolation methods for anaerobes in food. Int. J. Carman, RJ. (1997): Clostridium perfringens in spontaneous and Collie, R.E.; Kokai-Kun, JF. and McClane, B.A. (1998): Phenotypic Cornillot, E.B.; Saint-Joanis, B.; Daube, G.; Katayama, S.; Granum, Fernandez-Miyakawa M.E.; Marcellino, R. and Uzal, F.A. (2007): Food and Drug Administration 1998: Bacteriologic analytical manual, Guennadi, A.; Khoudoli, L; Porter, M.; Julian Blow, M. and Jason R.S. Hobbs, G.; Grouther, GS. and Neaves, P. (1982): Detection-and Immerseel, F.V.; Buck, J.D.; Pasmans, F.; Huyghebaert, G.; Pasmans, Jihong, Li, and Bruce, A. McClane (2006): Further Comparison of Koburger, J.A.. (1975): Understanding and teaching the most probable Assiut Vet, Med. J. Voi_53 No. 115 October 2007 Lin, ¥.T. and Labbe, R. (2003): Enterotoxigenicity and genetic Li, J. and McClane, B.A. (2006): Further comparison of temperature McClane, B.A. (2001) : Clostridium perfringens, In M. P. Doyle, L. R. Mead, G.C. (1992): Principles involved in the detection and enumeration Narayan, K.G. (1982): Food borne infection with Clostridium Pierson, M.D. and Reddy, N.R. (1988): Clostridium botulinum, Food Raju, D. and Sarker, M.R. (2005): Comparison of the levels of heat Ridell, J.; Bjorkroth, J; Eisgruber, H.; Schalch, B.; Stolle, A. and Sarker, M.R.; Carman, R.J. and McClane, B.A. (1999): Inactivation of Segner, W.P. (1979): Mesophilic aerobic spore forming bacteria in the Skjelkvale, R. and Uemura, T. (1977): Experimental diarrhea in human Songer, J.G. (1996): Clostridial enteric diseases of domestic animals. Stern, B.H. and Batty, 1. (1975): Pathogenic clostridia. 1* E.D. butter 10 Assiut Vet. Med J. Vol 53 No. 115 October 2007 Varga, J.; Stirewalt, VL. and Melville, S.B. (2004): The Cep proteins is Vernon, E.; Coyne, M;; Diane, J.; Sharon, JR. and Edward, P.R. (1996): Wen, O. and McClane, B.A. (2004): Detection of enterotoxigenic and enterotoxin gene (cpe)
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