Buzağı ishal olgularında Bovine Rotavirus (BRV) ve Bovine Coronavirus (BCoV) tespiti ve karakterizasyonu
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Date
2023-08-28
Authors
Ateş, Özer
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Neonatal döneminde en önemli mortalite sebebi olan buzağı ishalleri birçok faktöre bağlı olarak gelişmekte ve büyük ekonomik kayıplara sebep olmaktadır. Türkiye’ de bugüne kadar yapılan epidemiyolojik çalışmalarda, BRV ve BCoV’ un yeni doğan buzağılarda önemli oranda ishal olgularına neden olduğu gösterilmiştir. Bu çalışmada Türkiye’ nin farklı illerinden toplanan dışkı örneklerinde BRV ve BCoV’un rolü tespit edilmek istenmiştir. Projede kapsamında toplanan ve ishal semptomu gösteren bir aylık ve daha küçük yaştaki toplam 213 dışkı örneğinden 69 adedi (%32,39) BRV yönünden ve 17 adedi (%7,98) BCoV yönünden pozitif bulundu (Tablo4.2). Örneklerinin 82 adedinde (% 38,5) araştırılan viruslardan en az birisi (BRV ve/veya BCoV) tespit edilirken, 65 (%30,52) adet dışkı örneğinde tek BRV, 13(%6,10) adet dışkı örneğinde tek BCoV ve 4 (%1,88) adet dışkı örneğinde BRV ve BCoV ikili enfeksiyonu saptandı. Hücre kültüründe virus izolasyon çalışması sonucuMA-104 hücre hatında 2 adet (RV-36, RV-38) saha izolatı başarılı bir şekilde izole edildi. BRV antijen ELISA kiti ile pozitif tespit edilen 69 adet dışkı örneğinin 54 adedi RT-PCR yöntemi ile BRV yönünden pozitif olarak saptandı. BCoV antijen ELISA kiti ile pozitif tespit edilen 17 adet dışkı örneğinin 14 adedi Nested-PCR yöntemi ile BCoV yönünden pozitif olarak tespit edildi. 69 adet dışkı örneğine uygulanan G genotiplendirme RT-PCR sonuçlarına göre, G6 genotipinin tespit edilmesi amaçlanan genotipler arasında daha yüksek olmasına karşın belirgin olarak baskın bir genotip olmadığı tespit edildi. Ayrıca RT-PCR uygulamaları sonucu tespit edilmesi hedeflenen P genotipleri arasında da baskın bir genotipin bulunmadığı, P[5] ve P[11] genotiplerinin birbirine yakın düzeylerde görüldüğü saptandı. Tespit edilen BRV genotip kombinasyonları sonucu baskın olan genotipin G10P[11] olduğu belirlendi. Çalışmada 69 adet dışkı örneğindeki BRV segmentlerinin poliakrilamid jel üzerinde 4/2/3/2 şeklinde sıralandığı gösterilmiştir. Bu sonuçlar, Ag ELISA ve PCR ile BRV yönünden pozitif tespit edilen örneklerin, SDS-PAGE testi ile bant profillerine göre grup A'da olduğunu göstermektedir. Türkiye’ nin farklı illerinden yapılmış örnekleme çalışması sonrası elde edilen veriler doğrultusunda neonatal buzağı ishallerine sebep olabilen önemli viral etkenlerden BRV ve BCoV’ un yaygınlığı tespit edilmiştir ve işletmelerde kayıplara neden olabileceği gösterilmiştir.
Calf diarrhea, which is the most important cause of mortality in the neonatal period, develops due to many factors and causes great economic losses. In epidemiological studies conducted in Turkey to date, it has been shown that BRV and BCoV cause diarrhea in newborn calves at a significant rate. In this study, it was aimed to determine the role of BRV and BCoV in stool samples collected from different provinces of Turkey. Out of 213 stool samples, one month old and younger, collected within the scope of the project and showing diarrhea symptoms, 69 (32.39%) were found to be positive for BRV and 17 (7.98%) for BCoV. At least one of the investigated viruses (BRV and/or BCoV) was detected in 82 (38.5%) of the samples, 65 (30.52%) stool samples had single BRV, 13 (6.10%) stool samples had single BCoV and BRV and BCoV dual infection was detected in 4 (1.88%) stool samples. As a result of the virus isolation study in cell culture, 2 (RV-36, RV-38) field isolateswere successfully isolated from the MA-104 cell line. Fifty-four of 69 stool samples, which were detected positive with the BRV antigen ELISA kit, 54 were found to be positive for BRV by the RT-PCR method. Of the 17 stool samples that were detected positive with the BCoV antigen ELISA kit, 14 were detected as positive for BCoV by the Nested-PCR method. According to the G genotyping RT-PCR results applied to 69 stool samples, it was determined that although the G6 genotype was higher among the genotypes intended to be detected, it was not clearly a dominant genotype. In addition, it was determined that there was no dominant genotype among the P genotypes targeted to be detected as a result of RT-PCR applications, and P[5] and P[11] genotypes were observed at close levels.It was determined that the dominant genotype among the BRV genotyped samples was G10P[11]. In the study, it was shown that BRV segments in 69 stool samples were arranged as 4/2/3/2 on polyacrylamide gel. These results show that the samples detected positive for BRV by Ag ELISA and PCR are in group A according to the band profiles by SDS-PAGE test. The prevalence of BRV and BCoV, which are important viral agents that can cause neonatal calf diarrhea, has been determined in line with the data obtained after the sampling study conducted from different provinces of Turkey and it has been shown that they can cause losses in the enterprises.
Calf diarrhea, which is the most important cause of mortality in the neonatal period, develops due to many factors and causes great economic losses. In epidemiological studies conducted in Turkey to date, it has been shown that BRV and BCoV cause diarrhea in newborn calves at a significant rate. In this study, it was aimed to determine the role of BRV and BCoV in stool samples collected from different provinces of Turkey. Out of 213 stool samples, one month old and younger, collected within the scope of the project and showing diarrhea symptoms, 69 (32.39%) were found to be positive for BRV and 17 (7.98%) for BCoV. At least one of the investigated viruses (BRV and/or BCoV) was detected in 82 (38.5%) of the samples, 65 (30.52%) stool samples had single BRV, 13 (6.10%) stool samples had single BCoV and BRV and BCoV dual infection was detected in 4 (1.88%) stool samples. As a result of the virus isolation study in cell culture, 2 (RV-36, RV-38) field isolateswere successfully isolated from the MA-104 cell line. Fifty-four of 69 stool samples, which were detected positive with the BRV antigen ELISA kit, 54 were found to be positive for BRV by the RT-PCR method. Of the 17 stool samples that were detected positive with the BCoV antigen ELISA kit, 14 were detected as positive for BCoV by the Nested-PCR method. According to the G genotyping RT-PCR results applied to 69 stool samples, it was determined that although the G6 genotype was higher among the genotypes intended to be detected, it was not clearly a dominant genotype. In addition, it was determined that there was no dominant genotype among the P genotypes targeted to be detected as a result of RT-PCR applications, and P[5] and P[11] genotypes were observed at close levels.It was determined that the dominant genotype among the BRV genotyped samples was G10P[11]. In the study, it was shown that BRV segments in 69 stool samples were arranged as 4/2/3/2 on polyacrylamide gel. These results show that the samples detected positive for BRV by Ag ELISA and PCR are in group A according to the band profiles by SDS-PAGE test. The prevalence of BRV and BCoV, which are important viral agents that can cause neonatal calf diarrhea, has been determined in line with the data obtained after the sampling study conducted from different provinces of Turkey and it has been shown that they can cause losses in the enterprises.
Description
Keywords
Neonatal buzağı ishalleri, BRV, BCoV, Virus izolasyonu, ELISA, PCR, Genotiplendirme, PAGE, Neonatal calf diarrhea, Virus isolation, Genotyping
Citation
Ateş, Ö. (2023). Buzağı ishal olgularında Bovine Rotavirus (BRV) ve Bovine Coronavirus (BCoV) tespiti ve karakterizasyonu. Yayınlanmamış doktora tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.