Identification of Phenotypic and Genotypic Characteristics Diversity of Isolated Flavobacterium psychrophilum in Rainbow Trout Farms in Mediterranean Region

Year 2018, Volume: 14 Issue: 4, 265 - 275, 01.12.2018

Ahmet Tahir Ersoy Seçil Metin Ertan Emek Onuk

https://doi.org/10.22392/egirdir.400641

Cited By: 2

Abstract

In this study, it was aimed to investigate the phenotypic and genotypic characteristics with presence of
Flavobacterium psychrophilum in fourteen rainbow trout farms in Mediterranean Region. For this purpose,
sperm, eyed egg, egg, yolk-sac larvae and 0,2-5 g fry were sampled from commercial trout farms between
November 2012- March 2013. Isolation of potential F.psychrophilum from samples was made using
conventional microbiological methods and a total of 172 obtained strains. Enzyme activity of all isolates was
identified according to the API ZYM test system. As a result of the phenotypic tests, 65 isolates thought to be
F. psychrophilum were identified. PCR assay was used in molecular confirming of these strains. It was
determined that, 17 strains isolated from diseased fish (11 of them liver, 3 of them spleen, 3 of them kidney)
gave specific 971 bp band which was specific to F.psychrophilum. RAPD-PCR assay was used to identify the
clonal relationship between strains. F. psychrophilum isolates were yielded five different RAPD band patterns
(F1-F5) according to this method. This strains detected to be grouped into 1 cluster and according to similarity
coefficient %85 "unique" type (F1 the genotype). According to antibiotic sensitivity tests, isolates were found
to be sensitive to trimetroprim/sulfamethoxazole, clindamycin, ampicillin, tetracycline, enrofloxacin,
chloramphenicol, oxytetracycline, florfenicol and tobramycine. In conclusion, the findings from this study showed that the M13 primer used in RAPD-PCR can be used to determine the clonal relationship between F.
psychrophilum isolates 

Keywords

Oncorhynchus mykiss, Flavobacterium psychrophilum, identification, API ZYM, PCR

Ege ve Akdeniz Bölgesindeki Gökkuşağı Alabalıklarından İzole Edilen Flavobacterium psychrophilum Suşlarının Fenotipik Ve Genetik Farklılıklarının Belirlenmesi

Abstract

Bu çalışma ile Akdeniz bölgesinde bulunan 14 farklı gökkuşağı alabalığı (Oncorhynchus mykiss
Walbaum, 1792) işletmesinde Flavobacterium psychrophilum’un varlığı ile fenotipik ve genotipik
özelliklerinin araştırılması hedeflenmiştir. Bu amaçla Kasım 2012-Mart 2013 tarihleri arasında ticari alabalık
işletmelerinden sperm, yumurta, gözlenmiş yumurta, keseli larva ve 0.2-5 gr ağırlığındaki larvalardan örnekler
alınmıştır. Örneklerden olası F. pyschrophilum’un izolasyonu konvansiyonel mikrobiyolojik metotlar
kullanılarak yapılmış ve toplamda 172 adet izolat elde edilmiştir. Elde edilen tüm izolatların enzim aktivitesi
API ZYM test sistemine göre tespit edilmiştir. Yapılan fenotipik testler sonucunda F. psychrophilum olduğu
düşünülen 65 izolat belirlenmiştir. Bu izolatların moleküler olarak doğrulanmasında F. pyschrophilum spesifik
Polimeraz Zincir Reaksiyonu (PCR) metodu kullanılmıştır. Hasta balıklardan izole edilen 17 izolatın (11
karaciğer, 3 dalak ve 3 böbrek) F. psychrophilum’a özgü 971 bp’lik bant verdiği saptanmıştır. İzolatlar arası
klonal ilişkinin belirlenmesi amacıyla RAPD-PCR metodu kullanılmıştır. Bu metoda göre F. psychrophilum
izolatlarının beş farklı RAPD bant paterni (F1-F5) verdiği saptanmıştır. İzolatların % 85 benzerlik katsayısına
göre bir “unique” tip (F1 nolu genotip) ve bir küme içerisinde gruplandığı belirlenmiştir. Yapılan
antibiyogram testi sonucu, izolatların trimetroprim/sulfametakzol, klindamisine, ampisillin, tekrasiklin,
enrofloksasin, kloramfenikole, oksitetrasiklin, florfenikol ve tobramisin’e duyarlı olduğu belirlenmiştir. Sonuç
olarak, bu çalışmadan elde edilen bulgular RAPD-PCR'da kullanılan M13 primerinin F. psychrophilum
izolatları arasındaki klonal ilişkinin belirlenmesinde kullanılabilir olduğunu göstermiştir

Keywords

Oncorhynchus mykiss, Flavobacterium psychrophilum, identifikasyon, API ZYM, PCR

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