Önemli kanatlı solunum yolu viruslarının real-time polymerase chain reaction (rPCR) ile araştırılması
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Date
2021-06-02
Authors
Ardıçlı, Özge
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Bu çalışmada solunum yolu problemli tavuklarda Avian Coronavirus Infectious Bronchitis Virus (IBV), Infectious Laryngotracheitis Virus (ILTV), Avian Metapneumovirus (aMPV) ve Avian Reovirus (ARV) aranması ve IBV ve ILTV izolatlarının genetik karakterizasyonlarının yapılması hedeflendi. Bu amaçla Türkiye’de 15 ilde yer alan 45 yumurtacı ve 48 broyler kümeslerindeki tavuklara ait trakeal svablardan virus izolasyonu için ‘Specific Pathogen Free’ embriyolu yumurtalara ekim yapıldı. Bu yumurtaların koryoallantoik sıvılarında Newcastle Disease Virus (NDV) ve Avian Influenza Virus (AIV) varlığı Hemaglutinasyon ve Hemaglutinasyon İnhibisyon testleri ile saptandı. IBV, ILTV, aMPV ve ARV’ları ise Real-Time PCR (rPCR) testleri ile identifiye edildi. İdentifikasyon işlemleri sonucunda örneklerin hiçbirinde NDV ve AIV saptanmazken, yumurtacı örneklerinin 21’inde (%46,67) IBV, 10’unda (%22,22) ILTV, 6’sında (%13,33) aMPV ve 2’sinde (%4,44) ARV, broyler örneklerinin ise 28’inde (%58,33) IBV, 1’inde (%2,22) ILTV, 4’ünde (%8,33) aMPV ve 10’unda (%20,83) ARV varlığı tespit edildi. İncelenen yumurtacı ve broyler tavuklara ait örneklerin sırasıyla 5’inde (%11,11) ve 1’inde (%2,08) IBV+aMPV, 1’inde (%2,22) ve 5’inde (%10,42) IBV+ARV ikili olarak bulunurken, broyler örneklerin ise 2’sinde (%4,17) IBV+aMPV+ARV üçlü olarak saptandı. Birden fazla etken bulunan toplam örnek oranı %15,05 olarak hesaplandı. ILTV saptanan örneklerde başka bir virus belirlenmedi. Ayrıca yumurtacı ve broyler tavuk örneklerinin hiçbirinde ARV+aMPV ikili olarak saptanmadı. ILTV izolatlarının genetik karakterizasyonları sonucunda hepsinin saha suşu ve düşük virulensli oldukları kararına varıldı. IBV’ye ait S1 gen bölgesinin dizileme işlemi 5 örnekte başarılı oldu. Bu örneklerden 3’ünün 4/91 genotipi ile; diğer 2’sinin ise IS/1494/06 (Israel Var2) genotipi ile %99 oranında benzerlik gösterdiği belirlendi. Elde edilen sonuçların kanatlı endüstrisine faydalı olacağı düşünülmektedir.
In this study, it was aimed to search Avian Coronavirus Infectious Bronchitis Virus (IBV), Infectious Laryngotracheitis Virus (ILTV), Avian Metapneumovirus (aMPV), and Avian Reovirus (ARV) and to perform genetic characterization of IBV and ILTV isolates in chickens with respiratory problems. For this purpose, “Specific Pathogen Free” embryonated chicken eggs were inoculated for virus isolation from tracheal swabs of chickens in 45 layers and 48 broiler flocks located in 15 provinces in Turkey. The presence of Newcastle Disease Virus (NDV) and Avian Influenza Virus (AIV) in the chorioallantoic fluid of these eggs was detected by Hemagglutination and Hemagglutination Inhibition tests. IBV, ILTV, aMPV, and ARV were identified by Real-Time PCR (rPCR) tests. As a result of the identification process, NDV and AIV were not detected in any of the samples; concerning layer samples, the presence of 21 (46.67%) IBV, 10 (22.22%) ILTV, 6 (13.33%) aMPV, and 2 (% 4.44) ARV were detected, whereas, IBV in 28 (58.33%), ILTV in 1 (2.22%), aMPV in 4 (8.33%) and 10 (20.83%) ARV were detected in broiler samples. Of the layers and broiler chickens examined, respectively, IBV+aMPV in 5 (11.11%) and 1 (2.08%), IBV+ARV in 1 (2.22%), and 5 (10.42%) were found to be dual infections while IBV+aMPV+ARV were detected as triple in 2 (4.17%) broiler samples. The total sample rate with more than one agent was calculated as 15.05%. No other virus was detected in ILTV detected samples. In addition, ARV+aMPV was not detected as a dual infection in any of the layers and broiler chicken samples. Regarding the genetic characterization of ILTV isolates, it was concluded that they were all field strains and low virulence. Sequencing of the S1 gene region of IBV was successful in 5 samples. It was determined that 3 of these samples to 4/91 genotype and the other 2 were 99% similar to the IS / 1494/06 (Israel Var2) genotype. It is thought that the present results will be beneficial in the poultry industry.
In this study, it was aimed to search Avian Coronavirus Infectious Bronchitis Virus (IBV), Infectious Laryngotracheitis Virus (ILTV), Avian Metapneumovirus (aMPV), and Avian Reovirus (ARV) and to perform genetic characterization of IBV and ILTV isolates in chickens with respiratory problems. For this purpose, “Specific Pathogen Free” embryonated chicken eggs were inoculated for virus isolation from tracheal swabs of chickens in 45 layers and 48 broiler flocks located in 15 provinces in Turkey. The presence of Newcastle Disease Virus (NDV) and Avian Influenza Virus (AIV) in the chorioallantoic fluid of these eggs was detected by Hemagglutination and Hemagglutination Inhibition tests. IBV, ILTV, aMPV, and ARV were identified by Real-Time PCR (rPCR) tests. As a result of the identification process, NDV and AIV were not detected in any of the samples; concerning layer samples, the presence of 21 (46.67%) IBV, 10 (22.22%) ILTV, 6 (13.33%) aMPV, and 2 (% 4.44) ARV were detected, whereas, IBV in 28 (58.33%), ILTV in 1 (2.22%), aMPV in 4 (8.33%) and 10 (20.83%) ARV were detected in broiler samples. Of the layers and broiler chickens examined, respectively, IBV+aMPV in 5 (11.11%) and 1 (2.08%), IBV+ARV in 1 (2.22%), and 5 (10.42%) were found to be dual infections while IBV+aMPV+ARV were detected as triple in 2 (4.17%) broiler samples. The total sample rate with more than one agent was calculated as 15.05%. No other virus was detected in ILTV detected samples. In addition, ARV+aMPV was not detected as a dual infection in any of the layers and broiler chicken samples. Regarding the genetic characterization of ILTV isolates, it was concluded that they were all field strains and low virulence. Sequencing of the S1 gene region of IBV was successful in 5 samples. It was determined that 3 of these samples to 4/91 genotype and the other 2 were 99% similar to the IS / 1494/06 (Israel Var2) genotype. It is thought that the present results will be beneficial in the poultry industry.
Description
Keywords
Kanatlı solunum virusları, Tavuk, PCR, ILTV, IBV, Avian respiratory viruses, Chicken
Citation
Ardıçlı, Ö. (2021). Önemli kanatlı solunum yolu viruslarının real-time polymerase chain reaction (rPCR) ile araştırılması. Yayınlanmamış doktora tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.