Şükrü KIRKAN, Ergün Ö. GÖKSOY, Serten TEKBIYIK, Osman KAYA

Detection of yersinia ruckeri by PCR in rainbow trout (Oncorhynchus mykıss-Walbaum) hatchery farms in the West of Turkey

Bu araştırmada, Gökkuşağı Alabalığı çiftliklerindeki Yersinia ruckeri varlığı Polimeraz Zincir Reaksiyonu (PZR) metodu ile incelendi. Bu çalışmada Kızıl Ağız Hastalığı şüpheli 25 adet Gökkuşağı Alabalığı materyal olarak kullanıldı. 25 örneğin 8 (% 32)'inden Yersinia ruckeri identifıye edildi. 25 örneğin lO'unda PZR'da Yersinia ruckeri yönllnden pozitiflik saptandı. PZR pozitif Ürünlerin hepsinde, Yersinia ruckeri için spesifik 575 bp uzunluğunda bantlar elde edildi.

Türkiye' nin batısındaki Gökkuşağı alabalık yetiştirme çiftliklerinde Yersinia ruckeri' nin PZR kullanılarak tanımlanması

The presence of Yersinia ruckeri in rainbow trout hatchery farms was investigated by PCR Assay. Twenty-five rainbow trout, suspected of Enteric Redmouth Disease, were examined in this study. Yersinia ruckeri strains were identified from 8 out of 25 samples (32%). Ten of 25 (40%) rainbow trout samples showed positive reactions for Yersinia ruckeri in the PCR. All PCR positive samples have shown specific amplification bands of the 575-bp fragment for Yersinia ruckeri.

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1. Argenton, F., De Mas, S., Malocco, C, Dalla Valle, L., Giorgetti, G., Colombo, L.: Use of random DNA amplification to generate specific molecular probes for hybridization tests and PCRbased diagnosis of Yersinia ruckeri. Dis. Aquat. Organ., 1996; 24: 121-127.
2. Austin, B., Austin, D.A.: Bacterial fish pathogens: diseases in farmed and wild fish. 2nd edn. Ellis Horwood Limited, Chichester, UK., 1993; 208-227.
3. Busch, R.A., Lingg, A.J.: Establishment of an asymptomatic carrier state infection of enteric redmouth disease in rainbow trout {Salmo gairdneri). J. Fish Res. Board. Can., 1975; 32: 2429-2432.
4. Busch, R.A.: Enteric redmouth disease (Hagerman Strain). Mar. Fish Rev., 1978; 40: 42-51.
5. Ewing, W.H., Roos, A.J., Fahne, G.R.: Yersinia ruckeri sp. nov., the redmouth bacterium. Int.Syst. Bacteriol., 1978; 28: 37-44.
6. Fernandez-Garayzabal, J.F.: Development of a PCR assay for detection of Yersinia ruckeri in tissues of inoculated and naturally infected trout. Appl. Environ. Microbiol., 1999; 65: 46-350.
7. Fernandez-Garayzabal, J.F.: Molecular identification of pathogenic actinomycetes: a new approach. Microbiol. Sem., 1995; 11: 403-404.
8. Furones, M.D., Rodgers, C.J., Munn, C.B.: Yersinia ruckeri, the causal agent of enteric redmouth disease (ERM) in fish. Annu. Rev. Fish Dis., 1993; 3: 105-125.
9. Gendel, S.M.: Computational analysis of the specificity of 16S rRNA derived signature sequences for identifying food-related microbes. Food Microbiol., 1996; 13: 1-15.
10. Gibello, A., Blanco, M.M., Moreno, M.A., Cutuli, M.T., Domenech, A., Dominguez, L.,Fernandez-Garayzabal, J.F.: Development of a PCR assay for detection of Yersinia ruckeri in tissues of inoculated and naturally infected trout. Appl. Environ. Microbiol., 1999; 65: 346-350.
11. Greisen, K., Loeffelholz, M., Purohit, A., Leong, D.: PCR primers and probes for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid. J. Clin.Microbiol., 1994;32:335-351.
12. Holt, J.G., Krieg, N.R., Sneath, P.H.A., Stanley, J.T., Williams, T.S.: Bergey's Manual of Determinative Bacteriology. 9th ed„ William and Wilkins, Baltimore, Maryland, USA, 1994; pp.187.
13. Hunter, V.A., Knittel, M.D., Fryer, J.L.: Stress-induced transmission of Yersinia ruckeri infection from carriers to recipient steelhead trout Salmo gairdneri Richardson. J. Fish Dis., 1980;3: 467-472.
14. Ibrahim, A., Goebel, B.M., Liesack, W., Griffiths, M., Stackebrandt, E.: The phylogeny of the genus Yersinia based on 16S rDNA sequences. Fems. Microbiol. Lett., 1993; 114: 173-178.
15. Kirkan, S., Goksoy, E.O., Kaya, O.: The isolation of Yersinia ruckeri from rainbow trouts in Aydin region. The J. Pendik Vet. Microbiol., 2000; 31: 27-30
16. Kwok, S., Higuchi, R.: Avoiding false positives with PCR. Nature (London), 1989; 339: 237-238.
17. Lawson, P.A., Gharbia, S.E., Shah, H.N., Clark, D.R.: Recognition of Fusobacterium nucleatum subgroups Fn-1, Fn-2 and Fn-3 ribosomal RNA gene restriction patterns. FEMS. Microbiol. Lett.1989; 65: 41-46.
18. Leclercq, A., Wauters, G., Decallonne, J., El-Lioui, M., Vivegnis, J.: Usefulness of celluler fatty acid patterns for identification and pathogenicity of Yersinia species. Med Microbiol, lett. 1996; 5:182-194.
19. Lejeune, J.T., Rurangirwa, F.R.: Polymerase chain reaction for definitive identification of Yersinia ruckeri. J. Vet. Diagn. Investig., 2000; 12: 558-561.
20. Linde, H.J., Neubauer, II., Meyer, H., Aleksic, S., Lehn, N.: Identification of Yersinia species by the Vitek GNI card. J. Clin. Microbiol., 1999; 37: 211-214.
21. Mcintosh, D., Meaden, P.G., Austin, B.: A simplified PCR-based method for the detection of Renibacterium salmoninarum utilizing preparations of rainbow trout (Oncorhynchus mykiss,Walbaum) lymphocytes. Appl. Environ. Microbiol., 1996; 62: 3929-3932.
22. Romalde, J.L., Magarinos, B., Barja, J.L., Toranzo, A.E.: Antigenic and molecular characterization of Yersinia ruckeri: proposal for a new intraspecies classification. Syst. Appl.Microbiol., 1993; 16:411-419.
23. Rucker, R.: Redmouth disease of rainbow trout (Salmo gairdneri). Bull. Off. Int. Epi., 1966; 65:825-830.
24. Rys, P.N., Persing, D.H.: Preventing false positives: quantitative evaluation of three protocols for inactivation of polymerase chain reaction amplification products. J. Clin. Microbiol., 1993; 31:2356-2360.
25. Santos, Y., Romalde, J.L., Bandin, I., Magarinos, S., Nunez, S., Barja, J.L., Toranzo, A.E.:Usefulnes of the API-20E system for the identification of bacterial fish pathogens. Aquaculture,1993; 116: 111-120.
26. Saulnier, D., De Kinkelin, P.: Polymerase chain reaction primers for investigations on the causative agent of proliferative kidney disease of salmonids. J. Fish Dis., 1997; 20: 467-470.
27. Timur, G., Timur, M.: An outbreak of Enteric Redmouth Disease in farmed rainbow trout (O.mykiss) in Turkey. Bull. Euro. Ass. Fish Pathol., 1991; 11: 182-183.
28. Urdaci, M.C., Chakroun, C, Faurl, D., Bernardet, J.F.: Development of a polymerase chain reaction assay for detection and identification of the fish pathogen. Flavobacterium psychrophilum.Res. Microbiol., 1998; 149: 519-530.
29. White, T.J., Madej, R., Persing, D.H.: The polymerase chain reaction: clinical applications. Adv.Clin. Chem., 1992; 29: 161-196.
30. Zlotkin, A., Eldar, A„ Ghittino, C, Bercovier, H.: Identification of Lactococcus garviae by PCR.J. Clin. Microbiol., 1998; 36: 983-985.