Türkiye'deki Köy Tavuklarında İnfeksiyöz Laringotrakeitis Virüsünün Moleküler Tespiti ve Karakterizasyonu

Abstract

Gallid herpesvirüs I (GaHV1), infeksiyöz laringotrakeitisin (ILT) etiyolojik ajanıdır ve ILT, tavukların solunum yolu enfeksiyonları arasında yer alan önemli hastalıklardan biridir. Türkiye'de çok az çalışma yapılmış ve köy tavuklarında ILT'nin varlığına dair veri bulunmamaktadır. Çalışmada, köy tavuklarında PCR, RFLP ve sekans verileri kullanarak GaHV1'in tespitini ve karakterizasyonu amaçlandı. Toplamda 43 köy tavuğu işletmesinden alınan 163 trakeal swap örneğinin 5'inde (% 3,07) ILT enfeksiyonu pozitif bulundu. Sürü düzeyinde pozitiflik % 4,65 (2/43) idi. ICP4 geninin nükleik asit dizileri GenBank'taki ILTV dizileri ile karşılaştırıldığında benzerlik % 96.82 ile % 100 arasında değiştiği görüldü. Örneklerin sekans dizileri TCO suşları ile karşılaştırıldığında % 99.77-100 homoloji gözlemlendi. HgaI enzimi ile yapılan sanal RFLP analizinde, çalışmadaki suşlar TCO aşı suşuna benzer bir paterne sahip olarak karakterize edildi. Bu araştırma, köy tavuklarında GaHV1'in varlığını ve karakterizasyonunu ortaya koyan ilk çalışmadır. Türkiye'de ILT prevalansını tahmin etmek için büyük ölçekli çalışmalara ihtiyaç vardır. ILTV salgınlarını kontrol etmek için tavuklar izlenmeli ve yetiştiriciler aşılanmamış tavuklarla aşılanmış olanların temas etmesinden kaçınmalıdır.

Keywords

• GaHV1
• infeksiyöz laringotrakeitis
• köy tavuğu
• RFLP
• sekans

Molecular Detection and Characterization of Infectious Laryngotracheitis Virus in Backyard Chickens in Turkey

Abstract

Gallid herpesvirus I (GaHV1) is etiological agent of infectious laryngotracheitis (ILT) and ILT is one of the important disease that included in respiratory infections of chickens. Few studies have been conducted in Turkey and there is no data about the existence of ILT in backyards. The purpose of the study was to document the detection and characterization of GaHV1 in backyard chickens using PCR, RFLP and sequencing. Of the163 tracheal swap samples which were taken from 43 backyard flocks 5 (3.07%) were found to be positive for ILT infection. Positivity was 4.65 % (2/43) at flock level. The nucleic acid sequences of the ICP4 gene compared with ILTV sequences in GenBank and the level of identity differed from 96.82 to 100%. When the sequences of samples compared with TCO strains 99.77-100% homology was observed. The virtual RFLP analysis with the HgaI restriction enzyme characterized strains as having a pattern similar to the vaccine strain TCO. This is the first study that presents presence and characterization of GaHV1 in backyards. Large scale studies are needed to estimate prevalence of ILT in Turkey. Chickens should be monitored and growers should avoid to contacting vaccinated birds with non-vaccinated chickens, to control ILTV outbreaks.

Keywords

• backyard
• GaHV1
• infectious laryngotracheitis
• RFLP
• sequencing

References

1. Alaraji F, Hammadi H, Abed AA, Khudhair YI. Molecular detection and phylogenetic tree of infectious laryngotracheitis virus in layers in Al-Diwaniyah province, Iraq. Veterinar World. 2019; 12(4): 605-608.
2. Aras Z, Yavuz O, Gölen GS. Occurrence of infectious laryngotracheitis outbreaks in commercial layer hens detected by ELISA. Journal of Immunoassay and Immunochemistry. 2018; 39:2, 190-195. doi: 10.1080/15321819.2018.1428991.
3. Bayoumi M, El-Saied M, Amer H, Bastami M, Sakr EE, El-Mahdy M. Molecular characterization and genetic diversity of the infectious laryngotracheitis virus strains circulating in Egypt during the outbreaks of 2018 and 2019. Arch Virol. 2020; 165(3):661-670. doi:10.1007/s00705-019-04522-4.
4. Blacker H, Kirkpatrick N, Rubite A, O'Rourke D, Noormohammadi A. Epidemiology of recent outbreaks of infectious laryngotracheitis in poultry in Australia. Australian Veterinary Journal. 2011; 89(3):89-94. doi:10.1111/j.1751-0813.2010.00665.x.
5. Blakey J, Stoute S, Crossley B, Mete A. Retrospective analysis of infectious laryngotracheitis in backyard chicken flocks in California, 2007-2017, and determination of strain origin by partial ICP4 sequencing. J Vet Diagn Invest. 2019; 31(3):350-358. doi:10.1177/1040638719843574.
6. Creelan JL, Calvert VM, Graham DA, McCullough SJ. Rapid detection and characterization from field cases of infectious laryngotracheitis virus by real-time polymerase chain reaction and restriction fragment length polymorphism. Avian Pathol. 2006; 35:173-179.
7. Gowthaman V, Kumar S, Koul M, Dave U, Murthy T, Munuswamy P, Tiwari R, Karthik K, Dhama K, Michalak I, Joshi SK. Infectious laryngotracheitis: Etiology, epidemiology, pathobiology, and advances in diagnosis and control - a comprehensive review. The veterinary quarterly. 2020; 40(1):140–161. doi:10.1080/01652176.2020.1759845.
8. Gülaçtı I, Bulut H, Eröksüz Y, Çeribaşı AO. Outbreak of clinical infectious laryngotracheitis in Turkey. Vet. Rec. 2007; 160: 554–555. doi: 10.1136/vr.160.16.554.

9. Guy JS, Barnes HJ, Smith L. Increased virulence of modifed-live infectious laryngotracheitis vaccine virus following bird to-bird passage. Avian Dis. 1990; 35:348 https ://doi.org/10.2307/15911 88.
10. Guy JS, Garcia M. Laryngotracheitis. In: Disease of poultry, Ed; Saif YM, Fadly AM, Glisson JR, McDougald LR, Nolan LK, Swayne DE, 11th ed. Ames: Iowa State University Press, 2008; pp. 137-152 35.
11. Hughes CS, Williams RA, Gaskell RM, Jordan FTW, Bradbury JM, Bennett M, et al. Latency and reactivation of infectious laryngotracheitis vaccine virus. Arch Virol. 1991; 121:213-218.
12. Kaya BI and Akan M. First report of avian infectious laryngotracheitis infection in broiler breeders in Turkey. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2018; 65: 331-334.
13. Kearse M, Moir R, Wilson A, et al. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012; 28(12):1647-1649. doi:10.1093/bioinformatics/bts199.
14. King AMQ, Lefkowitz EJ, Mushegian AR. et al. Changes to taxonomy and the international code of virus classification and nomenclature ratified by the international committee on taxonomy of viruses. Arch Virol. 2018; 163, 2601–2631. https://doi.org/10.1007/s00705-018-3847-1.
15. Kotiw M, Wilks CR, May JT. The effect of serial in vivo passage on the expression of virulence and DNA stability of an infectious laryngotracheitis virus strain of low virulence. Vet Microbiol. 1995; 45:71–80. https://doi.org/10.1016/0378-1135(94)00115-D.
16. Menendez KR, Garcia M, Spatz S, Tablante NL. Molecular epidemiology of infectious laryngotracheitis: a review. Avian Pathol. 2014; 43(2):108–117.
17. Molini U, Aikukutu G, Khaiseb S. et al. Investigation of infectious laryngotracheitis outbreaks in Namibia in 2018. Trop Anim Health Prod. 2019; 51: 2105–2108 https://doi.org/10.1007/s11250-019-01918-x.
18. Oldoni I, García M. Characterization of infectious laryngotracheitis virus isolates from the US by polymerase chain reaction and restriction fragment length polymorphism of multiple genome regions. Avian Pathol. 2007; 36: 167-176.
19. Ou SC, Giambrone JJ. Infectious laryngotracheitis virus in chickens. World J Virol. 2012; 1(5): 142-149. doi: http://dx.doi.org/10.5501/wjv.v1.i5.142.
20. Thureen DR and Keeler CL. Psittacid herpesvirus 1 and infectious laryngotracheitis virus: Comparative genome sequence analysis of two avian alphaherpesviruses. J. Virol. 2006; 80:7863–7872.
21. Wanasawaeng W and Chansiripornchai N. Molecular classification of infectious laryngotracheitis virus from chick embryo origin vaccine, tissue culture origin vaccine and field isolates. Thai J. Vet. Med. 2010; 40:393–398.
22. Williams RA, Bennett M, Bradbury JM, Gaskell RM, Jones RC, Jordan FTW. Demonstration of sites of latency of infectious laryngotracheitis virus using the polymerase chain reaction. Journal of General Virology. 1993; 73: 2415–242.
23. World Organisation for Animal Health (OIE). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals 2019 Chapter 3.3.3. Avian infectious laryngotracheitis. OIE, Paris. Available at: https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.03.03_AVIAN_INF_LARYNGO.pdf. Accession date: 02.07.2020.
24. Yan Z, Li S, Xie Q, Chen F, Bi Y. Characterization of field strains of infectious laryngotracheitis virus in China by restriction fragment length polymorphism and sequence analysis. J Vet Diagn Invest. 2016; 28(1):46-49. doi:10.1177/1040638715618230.
25. Zhao Y, Kong C, Cui X, Cui H, Shi X, Zhang X, Hu S, Hao L, Wang Y. Detection of infectious laryngotracheitis virus by real-time PCR in naturally and experimentally infected chickens. PLoS One. 2013; 8(6):e67598.