Erzurum Yöresinde Koyunlarda Clostridium Perfringens Toksin Varlığının Toksin Nötralizasyon, ELI- SA ve Lateks Aglütinasyon Test Yöntemleri ile Araştırılması

Öz

Bu çalışmada, Erzurum yöresinde bulunan enterotoksemi şüpheli 70 adet koyuna ait bağırsak içeriğinin ELISA, toksin nötralizasyon ve lateks aglütinasyon testi (LAT) ile incelenerek; klostridial toksinlerin araştırılması ve tiplendirilmesi amaçlandı. Toksin testinde 25 (%35,7) örnekte C.perfringens yönünden pozitif sonuç alınırken, 45 (%64,3) adet örnek negatif olarak değerlendirildi. Bu test sonuçlarına göre, 14 örnek C.perfringens tip C, 2 örnek C.perfringens tip A, 9 örnek C.perfringens tip D olarak identifiye edildi. İncelenen örneklerin ELISA sonuçları değerlendirildiğinde 70 adet bağırsak içeriğinin 37 (%52,8) adeti C.perfringens yönünden pozitif bulunurken, 33 (%42,7) adet örnek negatif bulundu. Yapılan ELISA tiplendirilmesinde 12 adet numunede alfa toksin (tip A), 7 adet örnekte alfa, beta, epsilon toksin (tip B), 9 tanesinde alfa ve beta toksin (tip C) ve 9 adet örnek de alfa ve epsilon toksini (tip D) belirlendi. Lateks aglütinasyon sonuçlarında ise 2 örnek alfa toksin yönünden pozitif sonuç verirken, 16 örnekte epsilon toksini belirlendi. Sonuç olarak Erzurum ilinde enterotoksemi kontrolünde aşılamaların kontrollü, yaygın bir şekilde yapılmasının ve uygulanacak aşılarda başta tip A olmak üzere, tip C, tip D ile tip B’nin de bulundurulmasının gerekli olduğu belirlendi.

Anahtar Kelimeler

• Etlik Veteriner Mikrobiyoloji Dergisi
• Enterotoksemi
• Elisa
• Lateks Aglütinasyon
• Toksin Nötralizasyon
• Şenay SEYİTOĞLU
• Seyda CENGİZ
• Serap KILIÇ ALTUN
• Ömer Faruk KÜÇÜKKALEM
• İbrahim SÖZDUTMAZ
• 2012

Investigation of Clostridium Perfringens Toxins in Sheeps in the Region of Erzurum by Toxin Neutralization, ELISA and Lateks Agglütination Tests

Öz

Investigation and typing of clostridial toxins after ELISA, serum neutralization and latex agglutination tests (LAT) of intestinal contents from 70 enterotoxemia suspected sheeps in Erzurum region was aimed in this study. According to toxin neutralization results; 25 (35,7 %) and 45 (64,3 %) of the samples were determined as positive and negative, respectively. According to typing by toxin neutralization test; 14, 2 and 9 of the samples were typed as C, A and D, respectively. Thirty-seven (52,8 %) of the samples were positive for C.perfringens in the ELISA results. According to ELISA typing, 12 of the samples had alpha toxin (Type A), 7 of the samples had alpha, beta and gamma toxins (Type B), 9 of the samples had alpha and beta toxins (Type C), and 9 of the samples had alpha and epsilon toxins (Type D). According to latex agglutination test results, 2 of the samples had alpha and 16 of the samples had gamma toxins. As a result, the control of the province of Erzurum enterotoxemia, vaccination should be controlled and widely. Vaccines to be applied, especially in type A, type B, C and D must be present.

Kaynakça

1. Akan M, (2006). Clostridium Hastalıkları. Aydın N, Paracıkoğlu J. Eds. Veteriner Mikrobiyoloji (Bakteriyel Hastalıklar). İlke Emek Yayınları, Ankara. p. 73-86.
2. Gökçe Hİ, Genç O, Sözmen M, Gökçe G, (2007). Determination of Clostridium perfringens toxins types in sheeps with suspected enterotoxemia in Kars province. Turk. J. Vet. Anim. Sci. 31, 355-360.
3. Gökçe E, Ünver A, Erdoğan HM, (2010). İshalli neonatal kuzularda enterik patojenlerin belirlenmesi. Kafkas Univ Vet Fak Derg. 16, 717-722.
4. Idrissi AH, Ward GE, (1992). Evaluation of enzyme-linked immunosorbent assay for diagnosis of Clostridium perfringens enterotoxemias. Vet Microbiol. 31, 389-396.
5. Islam KBMS, Rahman MS, Ershauzzaman M, Taimur MJFA, Jang H, Song H, (2010). Detection of C.perfringens and its toxinotypes by ELISA from enterotoxaemic goats in Bangladesh. Korean J Vet Serv. 33, 37-44.
6. Kadra B, Guillou JP, Popoff M, Bourlioux P, (1999). Typing of sheep clinical isolates and identification of enterotoxigenic C.perfringens strains by classical methods and polymerase chain reaction (PCR). FEMS Immunol Med Microbiol. 24, 259-66.
7. Kalender H, Ertas HB, Cetınkaya B, Muz A, Arslan N, Kılıc A, (2005). Typing of isolates of Clostridium perfringens from healthy and diseased sheep by multiplex PCR. Vet Med Czech. 50, 439-442.
8. Koç R, Gökçe Hİ, (2007). Determination of the toxins and biotypes of C.perfringens in diarrhoeic calves in the Kars district of Turkey. Turk J Vet Anim Sci. 31, 207-11.

9. Laboratuvar Metot Birliği. Koruma-Kontrol Genel Müdürlüğü, Ankara. Erişim adresi: http://www.kkgm.gov. tr/genel/birimfaal.html
10. McClane B, Strouse RJ, (1983). Rapid detection of C.perfringens type A enterotoxin by Enzyme-Linked Immunosorbent Assay. J Clin Microbiol. 19, 112-115.
11. Miyakawa MEF, Uzal FA, (2005). Morphologic and physiologic changes induced by Clostridium perfringens type A alfa toxin in the intestine of sheep. AJVR. 66, 251-255.
12. Miyakawa MEF, Fisher DJ, Poon R, Sayeed S, Adams V, Rood J, McClane BA, Uzal FA, (2007). Both Epsilon toxin and Beta toxin are important for the lethal properties of C.perfringens type B isolates in the Mouse intravenous injection model. Infect Immun. 75, 1443-1452.
13. Moller K, Ahrens P, (1996). Comparison of toxicity neutralization- ELISA and PCR tests for typing of Clostridium perfringens and detection of the enterotoxin gene by PCR. Anaerobe. 2, 103-110.
14. Naylor RD, Martin PK, Barker LT, (1997). Detection of Clostridium perfringens alfa toksin by enzyme-linked immunosorbent assay. Res Vet Sci. 63, 101-112.
15. Niilo L, (1980). C.perfringens in animal disease: a review of current knowledge. Can Vet J. 21, 141-148.
16. Niilo L, (1986). Experimental production of hemorragic enterotoxemia by C.perfringens tip C in maturing lamb. Can J Vet Res. 50, 32-35.
17. Niilo L, (1987). Toxigenic characteristics of C.perfringens tip C in enterotoxemia in domestic animals. Can J Vet Res. 51, 224-228.
18. OIE (2004). Epsilon toxin of Clostridium perfringens. Institute for International Cooperation in animal biologics. Iowa State University.
19. Özcan C, Gürçay M, (2000). Elazığ ve çevresinde 1994- 1998 yılları arasında küçük ruminantlarda enterotoksemi ınsidensi. Turk J Vet Anim Sci. 24, 283-286.
20. Rosskopf-Streicher U, Volkers P, Werner E, (2003). Control of Clostridium perfringens vaccines using an indirect competitive ELISA for the epsilon toxin component. Pharmeuropa Bio. 2, 91-96.
21. Uzal FA, Niealsen K, Kelly WR, (1997). Detection of Clostridium perfringens type D epsilon antitoxin in serum of goats by competitive and indirect ELISA. Vet Microbiol. 51, 223-231.
22. Uzal FA, Kellay WR, Thomas R, Hornitzy M, Galea F, (2003). Comparison of four techniques for the detection of Clostridium perfringens type D epsilon toxin in intestinal contents and other body fluids of sheep and goats. J Vet Diagn Invest. 15, 94-99.
23. Watson PJ, Scholes SFE, (2009). Clostridium perfringens type D epsilon intoxication in one day old calves. Vet Rec. 164, 816-818.