Kültür Levreklerinde (Dicentrarchus labrax) Vibrio parahaemolyticus Enfeksiyonu

Bu çalışmanın amacı, Türkiye’nin Doğu Karadeniz Bölgesinde yüzer ağ kafeslerde yetiştirilen levrek balıklar (Dicentrarchus labrax, L. 1758)’nda görülen hastalık salgınlarından izole edilen suşların tanımlanması gerçekleştirilmiştir. Hastalık etkenlerine karşı antibiyotiklerin profilleri belirlenmiştir. Tipik hastalık semptomları gösteren levrek balıkların böbrek ve dalaklarından %1,5 tuz içeren triptik soy agara (%1,5 NaCl-TSA=T-TSA) ekimleri yapılmış ve 20±1°C de 48 saat soğutmalı etüvde inkübasyona bırakılmıştır. İzole edilen bakterileri tanımlamak için klasik mikrobiyolojik testler, API 20E hızlı test kitleri ve PZR metodu kullanılarak gerçekleştirilmiştir. Bakteri izolatları steril %1,5 tuzlu suda McFarland 0,5 standart bulanıklığında APİ 20E test kitlerine inokulasyonu yapılmıştır. PZR testi V. parahaemolyticus tespit etmek için 16S rRNA gen primerleri kullanılarak gerçekleştirilmiştir. İzolatların hepsi kanlı agarda (%5 koyun kanlı ilaveli) β homoliz oluşturduğu ve %7 NaCl içeren peptonlu suda iyi ürediği tespit edilmiştir. İzolatların hepsi mikrobiyolojik testler ve API 20E test kiti sonuçlarına göre V. parahaemolyticus olduğu tespit edilmiştir. Universal 16S rRNA primerleri kullanılarak yapılan PZR test sonucuna göre suşların %98 V. parahaemolyticus olduğu doğrulanmıştır. Antibiyogramı yapılan izolatlar sulphamethoxazole %100, ampicilline %84,4, eritromisine %71,9, oksitetrasiklin %62,5, trimetoprim-sulfametoksazol %56,3 ve streptomisin %46,9 dirençli, fakat suşların hepsi oksolinik asit, enrofloksasine ve florfenikol’e duyarlı olduğu tespit edildi

Anahtar Kelimeler:

Levrek, V. parahaemolyticus, API 20E, PZR

Infection of Vibrio parahaemolyticus in culture sea bass (Dicentrarchus labrax)

The aim of this study is to identify of the isolated bacteria strains from disease outbreaks in cultured sea bass (Dicentrarchus labrax L., 1758) in floating net cages in the Eastern Black Sea Region of Turkey. The profiles of antibiotics against disease agents were determined. The kidney and spleen of diseased sea bass that showed typical disease symptoms was streaked on the surface of tryptic soy agar (TSA) added with 1.5 % sodium chloride, and was incubated in the cooled incubator at 20±1°C for 48 hours. To identify of isolated bacteria was carried out by using conventional biochemical tests, the API 20E system tests and the PCR method. All isolates were inoculated in API 20E system test kits by adjusting to a turbidity matching a 0.5 McFarland standard in sterile 1.5% saline water. PCR assay was subjected to using a universal 16s rRNA gene primers to detect V. parahaemolyticus. All isolates showed β hemolysis on blood agar (with 5% sheep blood), and good growth in peptone water containing 7% NaCl. According to the 20E system and conventional biochemical test results were determined that all isolates were V. parahaemolyticus. All strains were confirmed as 98% V. parahaemolyticus by using PCR assay with 16S rRNA. Results of the testing susceptibility to antibiotics showed that V. parahaemolyticus isolates were resistant to 100% sulphamethoxazole, 84.4% ampicillin, 71.9% erythromycin, 62.5% oxytetracycline 56.3% trimethoprim-sulfamethoxazole and 46.9% streptomycin, but all strains were found susceptible to oxolinic acid, enrofloxacin and florphenicol.

Keywords:

Sea bass, V. parahaemolyticus, API 20E, PCR,

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