T. Tansel TANRIKUL, HAŞMET ÇAĞIRGAN, Erol TOKŞEN

Levrek' lerden (Dicentrarchus labrax., 1758) izole edilen Vibrio türlerinin API 20E yöntemiyle identifikasyonu

APİ 20E sistemi standartize edilmiş hızlı identifikasyon için önerilen bir test sistemidir.Bu minyatür multitest sistemi klinik laboratuvarlarda Gram (-) enterik bakterilerin identifikasyonu için geliştirilmiştir.Son yıllarda bu sistemin deniz ve tatlı su balığı patojenlerinin identifikasyonunda kullanımı gittikçe artmaktadır. Bu çalışmada da kültürü yapılan levreklerden (Dicentrarchus labrax L., 1758) izole edilen Vibrio türlerinin identifikasyonunda APİ 20E test kitleri kullanılmıştır. Ölmek üzere olan balıkların böbreklerinden sıvapla alınan numune %1.5 tuz içeren trypton soy agara (TSA) ve brain heart infusion agara (BHIA) ekilerek 21°C de 72 saate kadar inkube edildi. Koloniler thiosulphate citrate bile salts sucrose agara (TCBS) ekilerek saflaştırıldı. TCBS agarda üreyen koloniler yeniden %1.5 tuzlu TSA’ya geçildi. İdentifikasyon için APİ 20E test kitleri kullanıldı. Bakterilerin APİ 20E test kitlerine inokulasyonunda steril %1.5 tuzlu su kullanıldı.İzole edilen dokuz türden sekizi Vibrio anguillarum, biri ise Vibrio ordalii olarak identifiye edildi.Vibrio anguillarum türleri içersinde indol testi, inositol, sorbitol, amigdalin ve arabinozdan asit üretimi temel alınarak farklı türlerin olduğu görüldü.Vibrio ordalii ise Vibrio anguillarum’dan fenotipik testler ile kolaylıkla ayrılabildi.

Anahtar Kelimeler:

Vibrio, Vibrio ordalii, levrek, izolasyon, Dicentrarchus labrax, Listonella anguillarum, izolasyon teknikleri

Identification of isolated Vibrio sp. from sea bass (Dicentrarchus labrax L., 1758) using API 20E system

A standartdized and rapid identification system is offered by the API 20E system. Miniaturized multitest system was originally developed for the identification of Gram-negative enteric bacteria in clinical laboratories. During the last decade, this system has been increasingly used for the identification of marine and fresh water fish pathogens. In this study, API 20E strips were used for identification of Vibrio anguillarum and Vibrio ordalii which were isolated from cultured sea bass (Dicentrarchus labrax L., 1758). Samples were taken from kidney of moribund fish and spread over the surface of tryptone soya agar (TSA) which was supplemented with 1.5% sodium chloride. The plates were incubated at 25°C for 72 hours. Colonies were purified by three times streaking on thiosulphate citrate bile salts sucrose agar (TCBS). Pure colony was passaged to NaCl added TSA before to start identification with API 20E strips. Sterile 1.5% saline water was used inoculation of bacterium to API 20E strips. Eight of isolated nine strains were Vibrio anguillarum one strain was Vibrio ordalii. Arginin dihydrolase (ADH) gelatine degradation and acid production from glukoz, mannitol, saccharose were found positive; lysine decarboxylase (LDC), ornithine decarboxylase (ODC), H2S production, degradation of urea, acid production from rhamnose and melibiose were found negative in all V. anguillarum strains. Gelatine degradation, acid production from glucose, mannitol and sucrose were found positive in Vibrio ordalii but negative in ONPG, ADH, LDH, ODC, utilization of citrate, H2S production, degradation of urea, TDA, indole production, VP, acid production from inositole, sorbitole, rhamnose melibiose amygdaline and arabinise were found negative.

Keywords:

Vibrio, Vibrio ordalii, sea bass, isolation, Dicentrarchus labrax, Listonella anguillarum, isolation techniques,

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