Gökkuşağı Alabalığında Karşılaşılan Bazı Bakteriyel Hastalık Etkenlerinin Hızlı Teşhisi

Year 2018, , 23 - 27, 31.12.2018

İfakat Tulay Çağatay , Erkan Gümüş

https://doi.org/10.31797/vetbio.425026

Abstract

Bu
çalışmada, PCR yöntemi ile gökkuşağı alabalığında (Oncorhynchus mykiss) hastalığa neden olan beş farklı patojenin (Aeromonas salmonicida, Aeromonas hydophila, Yersinia ruckeri, Flavobacterium psychrophila ve Renibacterium salmoninarum) hızlı teşhis
edilebilmesi için beşyüz elli balık doku (solungaç, böbrek, ağız, deri ve
yüzgeç) örneğinden direk olarak saflaştırılan DNA’lar kullanılmıştır.
Çalışmamızdaki balık hastalık etkenlerini tespit edebilmek için beş farklı
hedef gen bölgesi (gryA, gyrB,
ompTS, n-DNA ve YER) kullanılmıştır ve bunların PCR reaksiyonları
gerçekleştirilmiştir. Çalışmamızda sonuç olarak, hastalıklara neden olan etken
bakteriler, dokulardan izole
edilen DNA’lardaki hedef genlerin PCR methodu kullanılarak tespit
edilmesiyle 3-4 saat içerisinde tanımlanmıştır. 

Keywords

PCR, Balık hastalığı

Rapid Detection of Some Bacterial Diseases in Rainbow Trout

Abstract

In this study, we used five hundred fifty
different tissue samples (gill, kidney, mouth, skin and finn) to detect five
pathogens (A. salmonicida, A. hydophila, Y. ruckeri, F. psychrophila
and R. salmoninarum)
in rainbow trout (Oncorhynchus
mykiss) with using direct PCR methods. Five targeted genes (gryA,
gyrB, ompTS, n-DNA and YER) have been chosen to detect the causative agents of
diseases. As a result, five infection agents of rainbow trout were identified
and determinated within 3-4 hours time by using direct PCR from fish tissues,
without any microbiological examination methods.

Keywords

PCR, Fish Diseases

References

• Austin, B., Austin, D.A. (2007). Bacterial Fish Pathogens Diseases of Farmed and Wild Fish. Springer, Praxis Publishing Chichester, UK.
• Balcazar, J.L., Vendrell, D., de Blas, I., Ruiz-Zarzuela, I., Gironés, O., Múzquiz, J.L. (2007). Quantitative detection of A.salmonicida in fish tissue by real-time PCR using self-quenched, fluorogenic primers. J. Med. Mic., 56(3), 323-8.
• Cerro, A., Marquez, I., Guıjarro, J.A. (2002). Cerro, A., Marquez, I., Guıjarro, J.A. (2002). Simultaneous detection of A.salmonicida. Appl. Envir. Mic., 68(10), 5177-5180.
• Chu, W.H., Lu, C.P. (2005). Multiplex PCR assay for the detection of pathogenic A.hydrophila. J. Fish Dis. 28(7), 437-441.
• Diler, Ö. Altun, S., Çalıkuşu, F., Diler, A. (2000). Gökkuşağı alabalığı (O. mykiss)’nin yaşadığı ortam ile ilişkili kalitatif ve kantitatif bakteriyel florası üzerine bir araştırma, Turkish J. Vet. Anim. Sci., 24, 251-259.
• Gibello, A., Blanco, M.M., Moreno, M.A., Cutuli, M.T., Domenech, A., Domínguez, L., Fernández-Garayzábal, J.F. (1999). Development of a PCR assay for detection of Y. ruckeri in tissues of inoculated and naturally infected trout. Appl. Envir. Mic., 65(1), 346-350.
• Gustafson, C.E., Thomas, C.J., Trust, T.J. (1992). Detection of A.salmonicida from fish by using PCR amplification of the virulence surface array protein gene. App. Envir.Mic., 58(12), 3816-25.
• Hidalgo, R., Magi, G.E., Balboa, S., Barja, J.L., Romalde, J.L. (2008).Development of a PCR protocol for the detection of A.salmonicida in fishby ampli. of the fstA gene. Vet. Mic., 128(3), 386-394.
• Khushiramani, R., Girisha, S.K., Karunasagar, I., Karunasagar, I.,(2007). Cloning and expression of an outer membrane protein ompTS ofA.hydrophila and study of immunogenicity in fish. Pro. Exp. Pur.51(2):303-7.
• Kingombe, C.I.B., D'Aoust, J.Y., Huys, G., Hofmann, L., Rao, M., Kwan, J. (2010). Multiplex PCR method for detection of three A. enterotoxin genes. Appl. Envir. Mic., 76, 425-433.
• Miriam, A., Griffiths, S.G., Lovely, J.E., Lynch, W.H. (1997). PCR and probe-PCR assays to monitor broodstock Atlantic salmon ovarian fluid and kidney for presence of DNA of the fish pathogen R.salmoninarum. J. Clin. Mic., 35(6), 1322-1326.
• Murcia, M.A.J., Soler, L., Saavedra, M.J., Chacón, M.R., Guarro, J., Stackebrandt, E. (2005). Phenotypic, genotypic, and phylogenetic discrepancies to differentiate A.salmonicida from A. bestiarum. In. Mic., 8(4), 259-269.
• Onuk, E.E., Ciftci, A., Findik, A., Durmaz, Y. (2010). Development and evaluation of a multiplex PCR assay for simultaneous detection of F.psychrophilum, Y.ruckeri and A.salmonicida in culture fisheries. J. Vet. Sci., 11(3), 235-241.
• Powell, M., Overturf, K., Hogge, C., Johnson, K. (2005). Detection of R.salmoninarum in chinook salmon, O. tshawytscha, using q-PCR. J. Fish Dis., 28(10), 615-622.
• Rhodes, L.D., Coady, A.M., .Deinhard, R.K. (2004). Identification of a msa gene in R.salmoninarum and the associated virulence phenotype. Appl.Envir.Mic., 70(11), 6488-94.
• Roozbahani, M.R., Bandehpour, M., Haghighi-Khiabanian-Asl, A.Abdollahi, H., Kazemi, B., (2009). PCR-Based detection of Y. ruckeriInfection in Rainbow Trout Fish. Asian J. Ani. Vet. Adv. 4(5):258-62.
• TUİK, 2017. Türkiye İstatistik Kurumu http://www.tuik.gov.tr. Alıntı tarihi: 2018
• Uma, A., Rebecca, G., Meena, S., Saravanabava, K. (2010). PCR Detection of putative aerolysin and hemolysin genes in an A.hydrophila isolate fromi Koi carp (Cyprinus carpio). Tamilnadu J. Vet. Anim. Sci., 6(1), 31-33.
• Yeh, H. Y., Shoemaker, C.A., Klesius P.H. (2006). Sensitive and rapid detection of F.columnare in channel catfish Ictalurus punctatus by a loop‐mediated isothermal amplification method. J. Appl. Mic., 100, 919-925.
• Yugueros, J., Temprano, A., Luengo, J.M., Naharro, G. (2001). Molecular cloning of Y.ruckeri aroA gene: a useful taxonomic tool. J. Fish Dis., 24(7), 383-90.