| The Identıfıcatıon Of Aeromonas Hydrophila Assocıated Wıth Hemorraghıc Septıcemıa In Freshwater Fısh By 16s Rdna Rflp And Rapd Analysıs.
The aim of this study was to identify Aeromonas hydrophila associated with hemorraghic septicemia in freshwater fish by rapid and sensitive 16S rDNA RFLP (Restriction Fragment Length Polymorphism) and RAPD (Randomly Amplified Polymorphic DNA) analysis instead of time-consuming conventional bacteriological methods.
For this purpose, 12 Aeromonas hydrophila strains isolated from diseased cultured rainbow trout and various aquarium fish were identified by conventional bacteriologic methods. The majority of the fish that examined had haemorrhages on the body surface, in the mouth and at the base of fins, pale gills, enlarged spleen and haemorrhages in the visceral organs.
DNA of isolated Aeromonas hydrophila strains and referance strain was extracted by using DNA Purification Kit (Roche Inc.) according to the manufacturer’s protocol. PCR for 16S rDNA RFLP analysis was performed by using specific primers. Enzymatic digestion were performed by incubating the amplication products with MboI and AluI at 37ºC for 3 hours. The RAPD analysis of Aeromonas hydrophila strains were performed using three primers. Aliquot of restriction reaction mixture and amplified RAPD DNA fragments were electrophoresed on a 2,5% agarose gel. After electrophoresis, gels were stained with ethidium bromide, examined and photographed under a UV transilluminator.
All strains showed the same RFLP pattern and identified as Aeromonas hydrophila. On the other hand, the RAPD profiles of Aeromonas hydrophila differed between isolates. This findings reveal genetic variety in Aeromonas hydrophila strains.
DELEN Derya ,
Danışman : Prof. Dr. Gülşen TİMUR
Anabilim Dalı : Su Ürünleri Yetiştiriciliği Anabilim Dalı
Programı (Varsa) : Hastalıklar Programı
Mezuniyet Yılı : 2007
Tez Savunma Jürisi : Prof. Dr. Gülşen TİMUR (Danışman)
Prof. Dr. Akın CANDAN
Doç. Dr. Süheyla KARATAŞ
Doç. Dr. Gülşen TİMUR
Yard. Doç. Dr. Tülay AKAYLI.
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