Molecular Characterization and Phylogenetic Analysis of Canine Parvovirus 2 in Dogs, Mersin Province, Turkey

Öz

Canine parvovirus type 2 (CPV-2) is a highly contagious and lethal viral disease with acute gastroenteritis in young dogs. After it first appeared in USA in 1978, continuous evolution of virus genome has arisen new genetic and antigenic variants such as CPV-2a, CPV-2b, new CPV-2a, new CPV-2b and CPV-2c throughout the world. The aim of the present study was to detect CPV-2 genetic variants from blood samples of clinically ill dogs and to characterize CPV-2 genetic variants circulating in Mersin province (Turkey), by partial sequence analysis of VP2 gene. A total of 20 blood samples were collected from dogs with gastroenteritis from local veterinary clinics between April 2017 to September 2017. Out of 20, 7 samples were positive for CPV-2 genomic DNA using conventional PCR. Amino acid comparisons showed that Ile-324 mutation was detected in the VP2 of clinical CPV-2b samples, as differ previously studies Turkish genetic variants. A phylogenetic tree constructed based on VP2 gene sequences indicated that CPV-2b sequences from Mersin province were related to Indian genetic variants. Moreover, CPV-2b sequences from Mersin have clustered separately from other Turkish CPV-2b genetic variants previously reported. The results show that CPV-2b genetic variant is circulating among in dogs in Mersin province.

Anahtar Kelimeler

• Canine parvovirus
• dog
• phylogenetic analysis
• VP2 gene

Türkiye’de Mersin İlinde Köpeklerde Canine Parvovirus 2’nin Moleküler Karakterizasyonu ve Filogenetik Analizi

Öz

Canine parvovirus 2 (CPV-2) genç köpeklerde akut kanlı gastroenteritis ile seyreden oldukça bulaşıcı ve ölümcül bir viral hastalıktır. 1978’de ABD’de ilk ortaya çıktıktan sonra, virus genomunun sürekli evrimi, tüm dünyada CPV2a, CPV-2b, new CPV-2a, new CPV-2b ve CPV-2c gibi yeni genetik ve antijenik varyantların ortaya çıkmasına yol açmıştır. Bu çalışmanın amacı, Mersin (Türkiye) ilinde klinik olarak hasta köpeklerin kan örneklerinde CPV -2’yi tespit etmek ve dolaşımda olan CPV- 2 genetik varyatlarının karakterizasyonunu sağlamaktır. Bunun için 2017 Nisan ve 2017 Eylül ayları arasında, yerel kliniklerden 20 adet kan örneği gastroenteritisli köpeklerden toplandı. 20 örneğin 7’si PZR ile CPV- 2 yönünden pozitif olarak test edildi. Klinik CPV-2b örneklerinin VP2’sindeki amino asit karşılaştırmaları sonucunda daha önce çalışılan Türk soylarından farklı olarak Ile-324 mutasyonu tespit edildi. VP2 geni dizinlerinden köken alarak yapılan filogenetik ağaç, Mersin ilinden gelen CPV-2b dizilerinin Hindistan genetik varyantlarıyla ilişkili olduğunu göstermiştir. Aynı zamanda Mersin ilinden elde edilen CPV-2b dizinleri daha önce tespit edilen Türk genetik varyantlarından farklı bir dal şekillendirmiştir. Bu sonuçlar Mersin ilinde CPV-2b genetik varyantının köpekler arasında dolaştığını göstermiştir.

Anahtar Kelimeler

• Canine parvovirus
• köpek
• filogenetik analiz
• VP2 gen

Kaynakça

1. 1. Apaa TT, Daly JM, Tarlinton RE, (2016). Canine parvovirus (CPV- 2) variants circulating in Nigerian dogs. Vet Rec Open. 3, 1 -4. 2. Buonavoglia C, Martella V, Pratelli A, Tempesta M, Cavalli A, Buonavoglia D, Bozzo G, Elia G, Decaro N, & Carmichael L, (2001). Evidence for evoluation of canine parvovirus type 2 in Italy. J Gen Virol. 82, 3021 - 3025. 3. Charmichael LE, (1994). Canine parvovirus type -2.An evolving pathogen of dogs.Ann Vet Med. 135, 459- 464. 4. Decaro N, Martella V, Desario C, Bellacicco AL, Camero M, Manna L, D’Aloja D, Buonavoglia C, (2006). First detection of canine parvovirus type C in pups with haemorragica enteritis in Spain. J Vet Med. 53, 468-472. 5. Decaro N, Desario C, Addie DD, Martella V, Vieira JM, Elia G, Zicola A, Davis C, Thompson G, Thiry E, Truyen U, and Buonavoglia C, (2007). Molecular Epidemiology of Canine Parvovirus, Europe. Emerg Infect Dis. 13, 1222 - 124. 6. Dei Giudici S, Cubeddu T, Giagu A, Sanna G, Rocca S, Oggiano A, (2017). First molecular characterization of canine parvovirus strains in Sardinia, Italy. Arch Virol. 017, 3457-3. 7. Hong C, Decaro N, Desario C, Tanner P, Pardo CM, Sanchez S, Buonavoglia C, Saliki TJ, (2007). Occurrence of canine parvovirus type 2c in the United States. J Vet Diagn Invest. 19, 535-539. 8. Horiuchi M, Goto N, Ishiguro M, Shinagawa J,(1994). Mapping of determinants of the host range fort he canine cells in the genome of the canine parvovirus using canine parvovirus/mink enteritis virus chimeric viruses.J Gen Virol.75, 1319 - 1328. 9. Lin CN, Chien CH, Chio MT, Chueh LL, Hung MY, Hsu HS, (2014). Genetic characterization of type 2a canine parvovirus from Twaiwan reveals the emergence of an ıle324 mutation in VP2. Virol J. 11, 39. 10. Jeoung S, Ahn S, Kim D, (2008). Genetic analysis of VP2 gene canine parvovirus isolates in Korea. J Vet Med Sci. 70, 719 - 722. 11. Ntafis V, Xylouri E, Kalli I, Desario C, Mari V, Decaro N, Buonavoglia C,(2010). Characterization of Canine parvovirus 2 variants circulating in Greece J Vet Diagn Invest. 22, 737-740. 12. Martella V, Decaro N, Elia G, Buonavoglia C, (2005). Surveillance activity for canine parvovirus in Italy. J Vet Med. 52, 312-315. 13. Meers J, Kyaw-Tanner M, Benzink Z, Zwijnenberg R, (2007). Genetic analysis of canine parvovirus from dogs in Australia. Aust Vet J. 85, 392-396. 14. Miranda C, and Thompson G, (2016). Canine parvovirus: the worldwide occurrence of antigenic variants. J Gen Virol, 97, 2043 - 2057. 15. Mittal M, Chakravarti S, Mohapatra KJ, Chug KP, Dubey R, Upmanuyu V, Narwal PS, Kumar A, Churamani CP, Kanwar SN,(2014). Molecular typing of canine parvovirus strains circulating from 2008 to 2012 in an organized kennel in India reveals the possibility of vaccination failure. Infec Genet Evol. 23, 1- 6. 16. Mukhopadhyay HK, Matta SL, Amsaveni S, Antony PX, Thanislass J, Pillai RM, (2014). Phylogenetic analysis of canine parvovirus partial VP2 gene in India. Virus Genes. 48, 89 - 95. 17. Raj MJ, Mukhopadhyay KH, Thanislass J, Antony XP, Pillai MR, (2010). Isolation, molecular characterization and phylogenetic analysis if canineparvovirus. Infec Genet Evol. 10, 1237 - 1241. 18. Soma T, Taharaguchi S, Ohinata T, Ishii H, Hara M, (2013). Analysis of the VP2 protein gene of canine parvovirus strains from affected dogs in Japan. Res Vet Sci. 94, 368 - 371. 19. Tamura K, Stecher G, Peterson D, Filipski A, and Kumar S, (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol. 30, 2725 - 2729. 20. Timurkan M, Oguzoglu T, (2015). Molecular characterization of canine parvovirus (CPV) infection in Turkey. Vet Ital. 51, 39 - 44. 21. U. Truyen, (2006). Evolution of canine parvovirus-a need for new vaccines? Vet Microbiol. 117, 9-13 22. Yesilbag K, Yılmaz Z, Ozkul A, & Pratelli A, (2007). Aetiological role of viruses in puppies with diarrhoea. Vet Rec. 161, 169 - 170. 23. Yılmaz Z, Pratelli A, &Torun S, (2005). Distribution of antigen types of canine parvovirus type 2 in dogs with hemorrhagic enteritis in Turkey. Turk J. Vet Anim Sci. 29, 1073 - 1076. 24. Zhang R, Yang S, Zhang W, Zhang T, Xie Z, Feng H, Wang S, Xia X, (2010). Phylogenetic analysis of the VP2 gene of canine parvoviruses circulating in China.Virus Genes. 40, 397- 402. 25. Zhao Z, Liu H, Ding K, Peng C, Xue Q, Yu Z, Xue Y, (2016). Occurance of canine parvovirus in dogs from Henan province of China. Vet Res. 12, 138 - 145. 26. Zienius D, Lelesius R, Kavaliauskis H, Stankevicius A, Salomskas A, (2016). Pol J Vet Sci. 19, 187-196. 27. Zourkas E, Ward MP, Kelman M, (2015). Canine parvovirus in Australia: A comperative study of reported rural and urban cases. Vet Microbiol. 121, 198 - 203.