Canine Coronavirusun Tespiti ve Kısmi Membran Gen Dizisi Temelli Moleküler Karakterizasyonu

Yıl 2024, Cilt: 35 Sayı: 1, 27 - 31, 29.03.2024

Zeynep Karapınar Mehmet Ozkan Timurkan

https://doi.org/10.36483/vanvetj.1302205

Öz

Köpeklerin Koronavirus (CCoV) enfeksiyonu tüm dünyada yaygın olarak görülmekte ve gastroenteritis bulguları ile seyretmektedir. Sekonder etkenlerle komplikasyon sonucunda enfeksiyon, özellikle yavru köpeklerde ölümle sonuçlanabilir. Dışkı ile atılan virus, dışkının yeme, suya ve çevreye bulaşmasıyla indirekt olarak yayılır. Yapılan bu çalışmada CCoV enfeksiyonunun ortaya konması, enfeksiyona ilişkin güncel moleküler bilgilerin elde edilmesi amaçlanmıştır. Ayrıca, bölgede dolaşan CCoV suşlarının M (membran proteini) genine dayalı moleküler karakterizasyonu da yapılmıştır. Çalışma materyalini, gastroenteritis semptomları gösteren 12 adet köpeğe ait gaita örneği oluşturdu. PCR ürünleri saflaştırdıktan sonra sekans analizine tabi tutuldu ve GenBank veri tabanından sağlanan farklı referans CCoV izolatları ile karşılaştırılarak filogenetik ağaç oluşturuldu. Filogenetik analiz sonucu 5 pozitif örnekten, 1 adedinin CCoV-I, 4 adet örneğin ise CCoV-IIa olduğu tespit edildi. Elde edilen suşların kendi aralarında %85.4-%97.7 oranında, GenBank veri tabanından elde edilen diğer suşlarla aralarında %82,7-%98 oranında benzerlik olduğu tespit edildi. Çalışma sonucunda, Balıkesir bölgesinde sirküle olan CcoV’ye yönelik güncel moleküler bilgiler elde edilmiştir. Bu çalışma ile, Türkiye’de CCoV enfeksiyonunun varlığına ve moleküler epidemiyolojisine ilişkin yapılacak yeni araştırmaların aşı çalışmalarına ve enfeksiyonun kontrolüne önemli katkılar sağlayacağı düşünülmektedir.

Anahtar Kelimeler

Köpek koronavirus, filogenetik analiz, PZR.

Proje Numarası

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Detection and Molecular Characterization of Canine Coronavirus Based on Partial Membrane Gene Sequences

Öz

Canine coronavirus (CCoV) infection in dogs is common all over the world and progresses with gastroenteritis findings. Infection as a result of complications with secondary factors may result in death, especially in puppies. The virus, which is excreted in the feces, spreads indirectly through the contamination of food, water, and the environment. This study, it was aimed at revealing the CCoV infection and obtaining current molecular information about the infection. In addition, molecular characterization of CCoV strains circulating in the region was made based on the M (membrane protein) gene. The study material consisted of stool samples from 12 dogs with gastroenteritis findings. The amplified PCR products were subjected to sequence analysis and a phylogenetic tree was constructed by comparing them with different reference CCoV isolates from GenBank. In the phylogenetic tree, 1 of the 5 positive samples was determined to be CCoV-I, and 4 samples were determined to be CCoV-IIa. It was determined that the strains obtained were 85.4 - 97.7% similar among themselves and 82.7-98% similar to other strains obtained from GenBank. As a result of study, current molecular information about CCoV circulating in the Balıkesir region was obtained. With this study, it is thought that new research on the existence and molecular epidemiology of CCoV infection in Türkiye will make important contributions to vaccine studies and the control of infection.

Anahtar Kelimeler

Canine coronavirus, PCR, phylogenetic analysis.

Destekleyen Kurum

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Proje Numarası

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Teşekkür

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Kaynakça

• Akkutay-Yoldar ZA, Koç BT, Oguzoglu TC (2020). Phylogenetic analysis of partial membrane protein gene of canine coronaviruses detected in Turkey. Ankara Univ Vet Fak, 67 (3), 265-271.
• Ataseven VS, Ucar H, Akca Y (2005). Canine coronavirus antibodies in stray dogs. Indian Vet J, 82 (7), 782-783.
• Avci O, Bulut O, Yapici O, Hasircioglu S, Simsek A (2016). Canine coronavirus infection in dogs in Turkey: Virological and serological evidence. Indian J Anım Res, 50 (4), 565-568.
• Benetka V, Kolodziejek J, Walk K, Rennhofer M, Möstl K (2006). M gene analysis of atypical strains of feline and canine coronavirus circulating in an Austrian animal shelter. Vet Rec, 159 (6), 170-174.
• Binn LN, Lazar EC, Keenan KP, et al. (1974). Recovery and characterization of a coronavirus from military dogs with diarrhea. Proc Annu Meet U S Anim Health Assoc, 78, 359-66.
• Buonavoglia C, Decaro N, Martella V et al. (2006). Canine coronavirus highly pathogenic for dogs. Emerg Infect Dis, 12 (3), 492-494.
• Brownlie J, Whittaker G (2017). Coronaviridae. In; Fenner’s veterinary virology 5. Edition, Elsevier Inc, United Kingdom.
• Decaro N, Buonavoglia C (2008). An update on canine coronaviruses: viral evolution and pathobiology. Vet Microbiol; 132:221–34.
• Decaro N, Mari V, Campolo M et al. (2009). Recombinant canine coronaviruses related to transmissible gastroenteritis virus of Swine are circulating in dogs. J Virol, 83 (3), 1532-1537.
• Decaro N, Buonavoglia C (2011). Canine coronavirus: not only an enteric pathogen. Vet Clin N Am-Small, 41 (6), 1121-1132.
• Decaro N, Lorusso A (2020). Novel human coronavirus (SARSCoV-2): A lesson from animal coronaviruses. Vet Microbiol, 244, 1-18.
• de Vries AAF, Horzinek MC, Rottier PJM, de Groot RJ (1997). The Genome Organization of the Nidovirales: Similarities and Differences between Arteri-, Toro-, and Coronaviruses. Semin Virol, 8 (1), 33-47.
• Doğan F, Köse Sİ (2022). Alt solunum yolu enfeksiyonu olan köpeklerde canine coronavirusun tespiti ve moleküler karakterizasyonu. Vet Hek Der Derg, 93 (2), 124-132.
• Dong B, Zhang X, Bai J et al. (2022). Epidemiological investigation of canine coronavirus infection in Chinese domestic dogs: A systematic review and data synthesis. Prev Vet Med, 209, 105792.
• Gür S, Civelek T (2007). Vaka Raporu: Bir köpek barınağında Kanin Koronavirus varlığının serolojik olarak araştırılması. Eurasian J Vet Sci, 21, 103-106.
• Gür S, Gençay A, Doğan N (2008). A Serologic investigation for canine corona virus infection in individually reared dogs in central Anatolia. J Fac Vet Med Univ Erciyes, 5, 67-71.
• Haake C, Cook S, Pusterla N, Murphy B (2020). Coronavirus infections in companion animals: virology, epidemiology, clinical and pathologic features. Viruses, 12 (9), 1023.
• Hall TA (1999). BioEdit: A User-Friendly Biological Sequence Alignment Editor and Analysis Program for Windows 95/98/NT. Nucleic Acids Symp Ser, 41, 95-98.
• He HJ, Zhang W, Liang J et al. (2020). Etiology and genetic evolution of canine coronavirus circulating in five provinces of China, during 2018–2019. Mic path, 145, 104209.
• Herrewegh AA, Smeenk I, Horzinek MC, Rottier PJ, De Groot RJ (1998). Feline coronavirus type II strains 79-1683 and 79-1146 originate from a double recombination between feline coronavirus type I and canine coronavirus. J Virol, 72 (5), 4508-4514.
• Lai MMC, Holmes KV (2001). Coronaviridae: the viruses and their replication. Fields virology. 4. Edition. Williams and Wilkins, Philadelphia: Lippincott.
• Lednicky JA, Tagliamonte MS, White SK et al. (2022). Isolation of a novel recombinant canine coronavirus from a visitor to Haiti: further evidence of transmission of coronaviruses of zoonotic origin to humans. Clin Infect Dis, 75 (1), e1184-e1187.
• Li N, Bai Y, Liu R et al. (2023). Prevalence and genetic diversity of canine coronavirus in northeastern China during 2019–2021. Comp Immunol Microbiol Infect Dis, 94, 101956.
• Ozan E, Tamer C (2020). Evcil hayvanların önemli coronavirüs enfeksiyonları. Avrasya Sağlık Bilim Derg, 3, 102-110.
• Pratelli A, Tempesta M, Greco G, Martella V, Buonavoglia C (1999). Development of a nested PCR assay for the detection of canine coronavirus. J Virol Methods, 80 (1), 11-15.
• Pratelli A, Martella V, Elia G et al. (2001). Severe enteric disease in an animal shelter associated with dual infections by canine adenovirus type 1 and canine coronavirus. J Vet Med, 48, 385-392.
• Pratelli A, Martella V, Pistello M et al. (2003). Identification of coronaviruses in dogs that segregate separately from the canine coronavirus genotype. J Virol Methods, 107 (2), 213-222.
• Pratelli A (2006). Genetic evolution of canine coronavirus and recent advances in prophylaxis. Vet Res, 37 (2), 191-200.
• Pratelli A, Tempesta M, Elia G et al. (2022). The knotty biology of canine coronavirus: A worrying model of coronaviruses' danger. Res Vet Sci, 144, 190-195.
• Radford AD, Singleton DA, Jewell C, et al. (2021). Outbreak of severe vomiting in dogs associated with a canine enteric coronavirus, United Kingdom. Emerg Infect Dis, 27 (2), 517.
• Singhal T (2020). A review of coronavirus disease-2019 (COVID-19). Indian J Pediatr, 87 (4), 281-286.
• Soma T, Ohinata T, Ishii H et al. (2011). Detection and genotyping of canine coronavirus RNA in diarrheic dogs in Japan. Res Vet Sci, 90 (2), 205-207.
• Tamura K, Stecher G, Kumar S (2021). MEGA11: Molecular Evolutionary Genetics Analysis version 11. Mol Biol Evol, 38, 3022-3027.
• Timurkan MO, Aydin H, Dincer E, Coskun N (2021). Molecular characterization of canine coronaviruses: an enteric and pantropic approach. Arch Viro, 166 (1), 35-42.
• Tizard IR (2020). Vaccination against coronaviruses in domestic animals. Vaccine, 38 (33), 5123-5130.
• Yeşilbağ K, Yılmaz Z, Torun S, Pratelli A (2004). Canine coronavirus infection in Turkish dog population. J Vet Med, 51, 353-355.