Isolation of Clostridium perfringens from Chickens and Detection of the Alpha Toxin Gene by Polymerase Chain Reaction (PCR)

This study was carried out to isolate Clostridium perfringens from chickens and to detect the gene encoding the alpha toxin produced by all types of C. perfringens by polymerase chain reaction (PCR). Intestinal contents of 160 slaughtered chickens from 8 different farms in Elazığ province were analyzed. C. perfringens was isolated from 8 (5%) of the samples. DNA samples extracted from suspected isolates grown on selective agar were amplified by PCR using a pair of primers derived from the alpha toxin gene. All of 8 suspected isolates were found to be C. perfringens by conventional methods and PCR. Isolates were typed by the toxin neutralization test. Of the 8 isolates, 6 were type A and 2 could not be typed. Since C. perfringens type A is associated with food poisoning in humans, the isolation of C. perfringens type A from chickens in this study is important for public health.

Isolation of Clostridium perfringens from Chickens and Detection of the Alpha Toxin Gene by Polymerase Chain Reaction (PCR)

This study was carried out to isolate Clostridium perfringens from chickens and to detect the gene encoding the alpha toxin produced by all types of C. perfringens by polymerase chain reaction (PCR). Intestinal contents of 160 slaughtered chickens from 8 different farms in Elazığ province were analyzed. C. perfringens was isolated from 8 (5%) of the samples. DNA samples extracted from suspected isolates grown on selective agar were amplified by PCR using a pair of primers derived from the alpha toxin gene. All of 8 suspected isolates were found to be C. perfringens by conventional methods and PCR. Isolates were typed by the toxin neutralization test. Of the 8 isolates, 6 were type A and 2 could not be typed. Since C. perfringens type A is associated with food poisoning in humans, the isolation of C. perfringens type A from chickens in this study is important for public health.

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  • Arda, M., Minbay, A., Leloğlu, N., Aydın, N., Kahraman, M., Akay, Ö., Ilgaz, A., İzgür, M., Diker, K.S.: Özel Mikrobiyoloji. Epidemiyoloji, Bakteriyel ve Mikotik İnfeksiyonlar. Medisan Yayın Serisi No: 26. Medisan Yayınevi. Ankara, 1997
  • Götze, U.: Vorkomen von Clostridium perfringens bei lebenden masthöhnchen und miglichkeiten der kontamination bein schlachten. Fleischwirtschaft, 1976; 56: 231-235. 6. Latinovic, V.: Study of characteristics of Clostridium perfringens strains isolated from broilers with enteritis. Veterinaria Yugoslavia, 1983; 32: 267-275.
  • Al-Sheikhly, F., Al-Saieg, A.: Role of coccidia in occurrence of necrotic enteritis of chickens. Avian Dis., 1980; 24: 324-333.
  • Baba, E., Ikemeto, T., Fukata, T., Sasai, K., Arakawa, A., McDonald, L.R.: Clostridial population and the intestinal lesions in chickens infected with Clostridium perfringens and Eimeria necatrix. Vet. Microbiol., 1997; 54: 301-308.
  • Demirözü, K.: Sağlıklı tavukların ince bağırsak içeriklerinde Clostridium welchii mikroorganizmalarının aranması ve bunun tavuk enterotoksemileri bakımından öneminin saptanması. Pendik Vet. Kont. Araflt. Enst., 1989; 20: 15-34.
  • Fach, P., Guillou, J.P.: Detection by in vitro amplification of the alpha-toxin (phospholipase C) gene from Clostridium perfringens. J. Appl. Bacteriol., 1993; 74: 61-66.
  • Engström, B.E., Fermer, C., Lindberg, A., Saarinen, E., Baverud, V., Gunnarsson, A.: Molecular typing of isolates of Clostridium perfringens from healthy and diseased poultry. Vet. Microbiol., 2003; 9: 225-235.
  • Yamagashi, T., Sugitani, K., Tanishima, K., Nakamura, S.: Polymerase chain reaction test for differentiation of five toxin types of Clostridium perfringens. Microbiol. Immunol., 1997; 41: 295-299.
  • Baums, C.G., Schotte, U., Amtsberg, G., Goethe, R.: Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. Vet. Microbiol., 2004; 20: 11-16
  • Tschirdewahn, B., Notermans, S., Wernars, K., Untermann, F.: The presence of enterotoxigenic Clostridium perfringens strains in faeces of various animals. Int. J. Food Microbiol., 1991; 14: 175- 178.
  • Long, J. R.: Studies on necrotic enteritis in broiler chickens with emphasis on the role of Clostridium perfringens. Disser. Abs. Int., 1974; 35: 2503.
  • Craven, S.E., Stern, N.J., Barley, J.S., Cox, N.A.: Incidence of Clostridium perfringens in broiler chickens and their environment during production and processing. Avian Dis., 2001; 45: 887- 896.
  • Nadas, Ü.G., Berkoğlu, A., Alp, R.: Sağlıklı tavukların kan serumlarında Clostridium welchii alfa antitoksininin aranması ve bunun tavuk enterotoksemileri yönünden değerlendirilmesi. Pendik Hay. Hast. Araflt. Enst. Derg., 1991; 22: 94-106.
  • Alp, R.: Kanatlı yemlerinde ve yem ham maddelerinde Clostridium welchii aranması ve bunun tavuk enterotoksemileri yönünden önemi. Pendik Vet. Kont. Araflt. Enst. Derg., 1992; 23: 147-154.
  • Russo, T.A., Gorbach, S.L.: Anaerobic Infections: Sporoformers, Diagnostic Procedures for Bacterial Infections. Ed: Wentworth, B.B. Seventh Edition. American Public Health Association. Inc., Washington, 1987
  • Çetinkaya, B., Karahan, M., Atil, E., Kalin, R., De Baere, T., Vaneechoutte, M: Study of prevalence of caseous lymphadenitis in sheep and goats in the east of Turkey. Vet. Microbiol., 2002; 88: 75-83.
  • Kanakaraj, R., Haris, D.C., Soger, J.G., Bosworth, B.: Multiplex PCR assay for detection of Clostridium perfringens in feces and intestinal contents of pigs and swine feed. Vet. Microbiol., 1998; 63: 29-38.
  • Miwa, N., Nishina, T., Kubo, S., Atsumi, M.: Most probable number method combined with nested polymerase chain reaction for detection and enumeration of enterotoxigenic Clostridium perfringens in intestinal contents of cattle, pigs and chickens. J. Vet. Med. Sci., 1997; 59: 89-92.
  • Arslan, N., Öztürk, G., Kalender, H., Katı, M.N.: Elazığ bölgesindeki ishalli koyun ve kuzularda Escherichia coli, Campylobacter ve Clostridium perfringens’in izolasyon ve identifikasyonu üzerine çalıflmalar. Pendik Vet. Kont. Araflt. Enst. Derg., 1996; 27: 43-53.
  • Holfshagen, M., Stenwig, H.: Toxin production by Clostridium perfringens isolated from broiler chickens and capercaillies (Tetrao urugalls) with and without necrotizing enteritis. Avian Dis., 1992; 36: 837-843.
  • Lukinmaa, S., Takkunen, E., Siitonen, A.: Molecular epidemiology of Clostridium perfringens related to foodborne outbreaks of disease in Finland from 1984 to 1999. Appl. Environ. Microbiol., 2002; 68: 3744-3749.