Buzağıların Rotavirus Enfeksiyonunun Prevalansının Poliakrilamid Jel Elektroforez Yöntemi ile Araştırılması

Abstract

Sığır rotavirus enfeksiyonları yenidoğanların ölümlerine neden olduğundan yetiştiriciler için önemli ekonomik kayıplara sebep olmaktadır. Reovirales takımı, Sedoreoviridae ailesi ve Rotavirus genusu içerisinde sınıflandırılan sığır rotavirusları en çok VP7 ve VP4 gen bölgelerine göre G ve P genotipleri olarak sınıflandırılmaktadır. Bunun haricinde diğer bir major antijen olan VP6 genetik ve antijenik özellikleri temel alınarak 10 farklı tür (grup A-J) belirlenmiştir. Grup A rotaviruslar sığırlarda en fazla ishale neden olurken grup B ve C enfeksiyonları da bilinmektedir. Buzağıların korunması için sürü bazında rotavirus taraması yapılmalı ve sığırların enfeksiyon durumu ortaya konulmalıdır. Bu amaçla Atatürk Üniversitesi Veteriner Fakültesi Viroloji Anabilim Dalı envanterinde bulunan ishal semptomu olan 100 adet buzağıya ait gaita örneği kullanıldı. Virus varlığını tespit etmek için grup ayrımına imkân veren poliakrilamid jel elektroforezi (PAGE) kullanıldı. Elektroforez öncesi gaita örneklerine nükleik asit ekstraksiyonu işlemi yapıldı ve sonrasında hazırlanan poliakrilamid jele yüklenerek yürütüldü. Yürütülen örnekler gümüş nitrat boyanarak segment paternleri belirlenerek rotavirus varlığına bakıldı. İncelenen örneklerin 27’sinde pozitiflik saptanırken 5 örnek şüpheli ve 68 örnek negatif olarak belirlendi. Pozitif bulunan örneklerin segment paterni grup A ile uyumlu bulundu ve tümünün bu grup içerisinde sınıflandığı görüldü. Her ne kadar aynı grup içerisinde yer alsa da tespit edilen pozitif örneklerin 3 farklı elektroforetipe sahip olduğu tespit edildi. Sonuç olarak rotavirusların buzağı ishalleri etiyolojisinde halen önemli rolü olduğu, tespit edilen rotavirusların grup A içerisinde yer almakla beraber varyasyon gösterdiği ve bölgede yetiştiricilerin kontrol ve hijyen önlemlerine dikkat etmesi gerektiği tespit edilmiştir.

Keywords

• Poliakrilamid jel elektroforezi
• Rotavirüs enfeksiyonları
• Sığır
• Tanı

Project Number

THD-2021-9090

Investigation of the Prevalence of Rotavirus Infection in Calves using Polyacrylamide Gel Electrophoresis

Abstract

Bovine rotaviruses cause loss of calves and cause great financial losses to breeders. Bovine rotaviruses, which are classified in the Reovirales order, Sedoreoviridae family and Rotavirus genus, are mostly classified as G and P genotypes according to VP7 and VP4 gene regions. In addition, 10 different species (group A-J) have been identified according to genetic and antigenic properties of another major antigen, VP6. Group A rotaviruses are the most common cause of diarrhea in calves, while group B and C infections are also known. For the protection of calves, rotavirus screening should be performed on a herd basis and the infection status of cattle should be revealed. For this purpose, stool samples of 100 calves with diarrhea symptoms in the inventory of Ataturk University, Faculty of Veterinary Medicine, Department of Virology were used. Polyacrylamide gel electrophoresis (PAGE), which allows the examination of segments of the genome, was used to check for the presence of the virus. Nucleic acid extraction was performed on the stool samples before electrophoresis and then extracts were loaded into the prepared polyacrylamide gel and run. The samples were stained with silver nitrate stain, segment patterns were determined, and the presence of rotavirus was analyzed. While 27 of the analyzed samples were positive, 5 samples were suspicious, and 68 samples were negative. The segment pattern of the positive samples was compatible with group A and all of them were classified in this group. Although they were in the same group, it was determined that the positive samples had 3 different electrophoretypes. As a result, it was determined that rotaviruses still have an important role in the etiology of calf diarrhea. Besides, the detected rotaviruses showed variation, although they were in group A, and breeders in the region should pay attention to control and hygiene measures.

Keywords

• Cattle
• Diagnosis
• Polyacrylamide gel electrophoresis
• Rotavirus infections

Supporting Institution

Bu araştırma Atatürk Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü tarafından “THD-2021-9090” nolu proje olarak desteklenmiştir.

Project Number

THD-2021-9090

Ethical Statement

Atatürk Üniversitesi Hayvan Deneyleri Yerel Etik Kurulu’ndan 19/02/2024 tarihli ve 2024/04-sayılı izin alınarak yapılmıştır.

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