KÜLTÜR GÖKKUŞAĞI ALABALIĞI (Oncorhynchus mykiss)’NDAN İZOLE EDİLEN GRAM-NEGATİF PATOJENLERİN LİPOPOLİSAKKARİT PROFİLLERİ

Yıl 2015, Cilt: 1 Sayı: 2, 80 - 89, 02.02.2015

Tülay Akaylı , Özgür Çanak , Çiğdem Ürkü

https://doi.org/10.3153/JAEFR15008

Öz

Bu çalışma, Türkiye’nin farklı bölgelerindeki kültür gökkuşağı alabalıkları
(Oncorhynchus mykiss)’ndan izole
edilen patojenik Gram-negatif bakterilerin lipo­polisakkarit (LPS) profillerinin
belirlenmesi amacıyla yürütülmüştür. Bu amaçla, hasta balık örneklerinden izole
edilen Vibrio anguillarum, Aeromonas hydrophi­la, A. schubertii,
Pseudomonas fluorescens ve Yersi­nia
ruckeri izolatlarından LPS numuneleri elde edil­miştir. LPS profilleri, Sodyum
Dodesil Sülfat-Poliakrilamid Jel Elektroforezi (SDS-PAGE) ve gümüş-nitrat
boyama metodu ile belirlenmiştir. Bu çalışmadan elde edilen sonuçlar, LPS
analizi çalışma­larının bu bakterilerin karakterizasyonunda ve değişik coğrafik
bölgelerden izole edilen suşlar arasındaki farklılıkların tespit edilmesinde
kullanılabilecek fay­dalı araçlar olduğunu göstermiştir.

Anahtar Kelimeler

Gökkuşağı alabalığı, O. mykiss, SDS-PAGE, LPS

LIPOPOLYSACCHARIDE PROFILES OF GRAM-NEGATIVE PATHOGENS RECOVERED FROM CULTURED RAINBOW TROUT (Oncorhynchus mykiss)

Öz

The aim of this
study was to determine the lipopoly­saccharide (LPS) profiles of pathogenic
Gram-negative bacteria recovered from cultured rainbow trout (Oncorhynchus mykiss) in different
regions of Turkey. For this purpose, LPS samples were obtained from 10
bacterial isolates of Vibrio anguillarum, Aer­omonas hydrophila, A.
schubertii, Pseudomonas fluo­rescens
and Yersinia ruckeri recovered from
diseased fish. LPS profiles were determined by using Sodium Dodecyl Sulphate
Polyacrilamide Gel Electrophoresis (SDS-PAGE) and silver-nitrate staining
method. Re­sults of the study demonstrated that LPS profiling is a useful tool
for characterization of these bacteria and geographic discrimination of their
isolates.

Anahtar Kelimeler

Rainbow trout, O. mykiss, SDS-PAGE, LPS

Kaynakça

• Acosta, F., Lockhart, K., Gahlawat, S.K., Real, F., Ellis, A.E. (2004): Mx expression in Atlantic salmon (Salmo salar L.) parr in response to Listonella anguillarum bacterin, lipopolysaccharide and chromosomal DNA. Fish & Shellfish Immunology, 17: 255-263.
• Akaylı, T. (2001). Kültür Çipura Balıklarında (Sparus auratus, L 1758) Vibriosis’in ELISA ve Bakteriyolojik Yöntemlerle Teşhisi. Doktora Tezi, İstanbul Üniversitesi Fen Bilimleri Enstitüsü.
• Akaylı, T., Çanak, Ö., Başaran, B. (2011a): Gökkuşağı alabalıklarında (Oncorhynchus mykiss Walbaum, 1792) görülen Aeromonas schubertii enfeksiyonu üzerine bir çalışma. Biyoloji Bilimleri Araştırma Dergisi, 4(1): 99-106.
• Akaylı, T., Çanak, Ö., Başaran, B. (2011b): Yavru kültür gökkuşağı alabalıklarında (Oncorhynchus mykiss Walbaum, 1792) görülen yeni bir Pseudomonas türü: Pseudomonas plecoglossicida. Biyoloji Bilimleri Araştırma Dergisi, 4(1): 107-111.
• Alsina, M., Martinez-Picado, J., Jofre, J., Blanch, A.R. (1994): A medium for presumptive identification of Vibrio anguillarum. Applied and Environmental Microbiology, 60(5): 1681-1683.
• Altınok, İ., Balta, F., Çapkın, E., Kayış, Ş. (2007): Diseases of rainbow trout caused by Pseudomonas luteola. Aquaculture, 273(4): 393-397.
• Altınok, İ., Kayış, Ş., Çapkın, E. (2006): Pseudomonas putida infection in rainbow trout. Aquaculture, 261(3): 850-855.
• Altun, S., Kubilay, A., Diler, Ö. (2010): Yersinia ruckeri suşlarının fenotipik ve serolojik özelliklerinin incelenmesi. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 16(Suppl-B): 223-229.
• Austin, B., Austin, D.A. (2012). Bacterial Fish Pathogens: Disease of Farmed and Wild Fish, 5th edition. Springer, New York. 978-94-007-4884-2.
• Baran, İ., Timur, M., Aydın, N., İstanbulluoğlu, E., Aydıntuğ, M.K. (1981): Çifteler-Sakaryabaşı balık üretim ve araştırma istasyonunda alabalıklarda (Salmo gairdneri irideus) görülen bakteriyel hemorajik septisemi hastalığı üzerine incelemeler. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 27: 467-473.
• Bastardo, A., Sierralta, V., Leon, J., Ravelo, C., Romalde, J.L. (2011): Phenotypical and genetic characterization of Yersinia ruckeri strains isolated from recent outbreaks in farmed rainbow trout Oncorhynchus mykiss (Walbaum) in Peru. Aquaculture, 317: 229-232.
• Boesen, H.T., Pedersen, K., Larsen, J.L., Koch, C., Ellis, A.E. (1999): Vibrio anguillarum resistance to rainbow trout (Oncorhynchus mykiss) serum: role of O-antigen structure of lipopolysaccharide. Infection and Immunity, 67(1): 294-301.
• Bos, M.P., Tomassen, J. (2004): Biogenesis of the Gram-negative bacterial outer membrane. Current opinion in Microbiology, 7: 610-616.
• Buller, N. (2004). Bacteria from Fish and other Aquatic Animals: a practical identification manual. CABI Publishing, Oxford, U.K., 0-85199-738-4.
• Chart, H., Trust, T. J. (1984): Characterization of the surface antigens of the marine fish pathogens Vibrio anguillarum and Vibrio ordalii. Canadian Journal of Microbiology, 30(5): 703-710.
• Çağırgan, H., Yüreklitürk, O. (1991): First isolation of Yersinia ruckeri from a rainbow trout farm in Turkey. In: EAFP 5th International Conference on Diseases of Fish and Shellfish 1991, Budapest, Book of Abstracts, pp.131.
• Çelikkale, M.S., Düzgüneş, E., Okumus, İ. (1999). Türkiye Su Ürünleri Sektörü Potansiyeli, Mevcut Durumu, Sorunları ve Çözüm Önerileri. İstanbul Ticaret Odası, Yay. No. 1999-2. İstanbul. 975-512-321-0.
• Davies, R.L. (1989). Biochemical and Cell-Surface Characteristics of Yersinia ruckeri in Relation to the Epizootiology and Pathogenesis of Infections in Fish. Doktora tezi, Institute of Aquaculture, University of Stirling.
• Diler, O., Altun, S. (1994): Gökkuşağı alabalıklarından (Oncorhynchus mykiss) hemorajik septisemi etkeni olarak izole edilen bazı Aeromonas hydrophila suşlarının biyokimyasal özelliklerinin belirlenmesi. Süleyman Demirel Üniversitesi Eğirdir Su Ürünleri Fakültesi Dergisi, 4: 169-178.
• Dooley, J.S., Trust, T.J. (1988): Surface protein composition of Aeromonas hydrophila strains virulent for fish: identification of a surface array protein. Journal of Bacteriology, 170(2): 499-506.
• Dooley, J.S.G., Lallier, R., Shaw, D. H., Trust, T.J. (1985): Electrophoretic and immunochemical analyses of the lipopolysaccharides from various strains of Aeromonas hydrophila. Journal of Bacteriology, 164(1): 263-269.
• Fulop, M., Manchee, R., Titball, R. (1995): Role of lipopolysaccharide and a major outer membrane protein from Francisella tularensis in the induction of immunity against tularemia. Vaccine, 13(13): 1220-1225.
• Hitchcock, P.J., Brown, T.M. (1984): Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. Journal of Bacteriology, 154: 269-277.
• Ingram, G.A. (1993). Diffusion in Gel Techniques. In: Stolen, J. S., Fletcher, T. C., Anderson, D. P., Roberson, B. S., van Muiswinkel, W. B., (Eds.), Techniques in Fish Immunology: FITC1, 2nd edition. SOS Publications, New Jersey. 0-9625505-0-7.
• Kayış, Ş., Çapkın, E., Balta, F., Altınok, İ. (2009): Bacteria in rainbow trout (Oncorhynchus mykiss) in the southern Black Sea region of Turkey. The Israeli Journal of Aquaculture – Bamidgeh, 61(4): 339-344.
• Kırkan, S., Göksoy, E.O., Kaya, O.(2003): Isolation and antimicrobial susceptibility of Aeromonas salmonicida in rainbow trout (Oncorhynchus mykiss) in Turkey hatchery farms. Journal of Veterinary Medicine, Series B, 50(7): 339-342.
• Kimberley, A. W., Macnair, N.G. (2004). Finfish and Shellfish Bacteriology Manual: Techniques and Procedures. Iowa State Press, Iowa, USA. 0-8183-1952-0.
• Knappskog, D.H., Rodseth, O.M., Slinde, E., Endersen, C. (1993): Immunochemical analyses of Vibrio anguillarum strains isolated from cod, Gadus morhua L., suffering from Vibriosis. Journal of Fish Diseases, 16: 327-338.
• Koebnik, R., Locher, K.P., Van Gelder, P. (2000): Structure and function of bacterial outer membrane proteins: barrels in a nutshell. Molecular Microbiology, 37(2): 239-253.
• Kokka, R.P., Lindquist, D., Abbott, S.L., Janda, J.M. (1992): Structural and pathogenic properties of Aeromonas schubertii. Infection and Immunity, 60(5): 2075-2082.
• Köprücü, S., Sarıeyyüpoğlu, M. (2010): Aeromonas hydrophila ile enfekte edilen gökkuşağı alabalığında (Oncorhynchus mykiss) histopatolojik bir araştırma. Fırat Üniversitesi Fen Bilimleri Dergisi, 22(1): 11-17.
• Kubilay, A., Timur, G. (2001): Yersinia ruckeri bakterini ile immunize edilen gökkuşağı alabalıklarında (Oncorhynchus mykiss) antikor üretiminin IFAT ve ELISA teknikleri ile saptanması. Turkish Journal of Veterinary and Animal Sciences, 25: 437-445.
• Kubilay, A., Altun, S., Diler, O., Ekici, S. (2008): Isolation of Flavobacterium columnare from cultured rainbow trout (Oncorhynchus mykiss) fry in Turkey. Turkish Journal of Fisheries and Aquatic Sciences, 8: 165-169.
• Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
• Lima De Faria, A. (1969). Handbook of Molecular Cytology. North-Holland Publications, Londra.
• Pazos, F., Santos, Y., Magarinos, B., Bandin, I., Nunez, S., Toranzo, A.E. (1993): Phenotypic characteristics and virulence of Vibrio anguillarum related organisms. Applied and Environmental Microbiology, 59(9): 2969-2976.
• Pedersen, K., Grisez, L., Houdt, R.V., Tiainen, T., Ollevier, F., Larsen, J.L. (1999): Extended serotyping scheme for Vibrio anguillarum with the definition and characterization of seven provisional O-serogroups. Current Microbiology, 38: 183-189.
• Roberts, R.J. (2012). Fish Pathology 4th edition. Wiley-Blackwell. 978-1444332827.
• Sakai, M. (1999): Current reseach status of fish immunostimulants. Aquaculture, 172: 63-92.
• Santos, Y., Pazos, F., Bandin, I., Toranzo, A.E. (1995): Analysis of antigens present in the extracellular products and cell surface of Vibrio anguillarum serotypes O1, O2 and O3. Applied and Environmental Microbiology, 61(7): 2493-2498.
• Sousa, J.A., Magarinos, B., Eiras, J.C., Toranzo, A.E., Romalde, J.L. (2001): Molecular characterization of Portuguese strains of Yersinia ruckeri isolated from fish culture systems. Journal of Fish Diseases, 24: 151-159.
• Solem, S.T., Jorgensen, J.B., Robertsen, B. (1995): Stimulation of respiratory burst and phagocytic activity in Atlantic salmon (Salmo salar L.) macrophages by lipopolysaccharide. Fish & Shellfish Immunology, 5: 475-491.
• Sorensen, U.B.S., Larsen, J.L. (1986): Serotyping of Vibrio anguillarum. Applied and Environmental Microbiology, 51(3): 593-597.
• Swain, P., Nayak, S.K., Sahu, A., Meher, P.K., Mishra, B.K. (2003): High antigenic cross-reaction among the bacterial species responsible for diseases of cultured freshwater fishes and strategies to overcome it for specific serodiagnosis. Comperative Immunology, Microbiology & Infectious Diseases, 26: 199-211.
• Tanrıkul, T.T. (2007): Vibriosis as an epizootic disease of rainbow trout (Onchorhynchus mykiss) in Turkey. Pakistan Journal of Biological Sciences, 10(10): 1733-1737.
• Tinsley, J.W., Lyndon, A. R., Austin, B. (2011): Antigenic and cross-protection studies of biotype 1 and biotype 2 isolates of Yersinia ruckeri in rainbow trout, Oncorhyncus mykiss (Walbaum). Journal of Applied Microbiology, 111:8-16.
• Timur, G., Korun, J. (2004): First outbreak of vibriosis in farmed rainbow trout (Oncorhynchus mykiss) in Turkey. İ. Ü. Su Urunleri Fakultesi Dergisi, 18(18): 1-9.
• Timur, G., Timur, M. (1991): An outbreak of enteric redmouth disease in farmed rainbow trout (O. mykiss) in Turkey. Bulletin of European Association of Fish Pathologists, 11(5): 182-183.
• Timur, G., Timur, M. (2003). Balık Hastalıkları. İstanbul Üniversitesi Su Ürünleri Fakültesi, Yayın No. 5, İstanbul. 975-404-699-9.
• Tsai, C.M., Frasch, C.E. (1982): A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Analytical Biochemistry, 119(1): 115-119.
• Türkiye İstatistik Kurumu (TUİK) [Online]: İstatistiksel Tablolar ve Dinamik Sorgulama - Kültür Balıkları Üretim Miktarı tablosu, http://www.tuik.gov.tr/PreTablo.do?alt_id=1005 [Ziyaret Tarihi: 6 Kasım 2013]