PERTANIKA JOURNAL OF TROPICAL AGRICULTURAL SCIENCE

Astroviruses have been associated with enteric and extra-intestinal disorders in many animal species, including chickens. Here, we describe the detection and characterisation of chicken astrovirus (CAstV) in broilers and its seroprevalence in broiler breeder flocks. Based on PCR protocol, viral confirmation was carried out on clinical tissue samples from broiler chickens suffering from uneven growth and poor performance. The tissues were molecularly detected for CAstV with differential diagnostic testing against the Newcastle disease virus, infectious bronchitis virus, avian nephritis virus, avian rotavirus, fowl adenovirus and avian reovirus. Polymerase gene-based phylogenetic analyses of the twenty samples detected positive for CAstV indicate they belong to Group I and are related to strains from the US, UK, India and Poland. From these 20 samples, CastV could be isolated from 3 samples upon inoculation in 5-day-old specific-pathogen-free (SPF) embryonated chicken eggs (ECE); virus-infected embryos showed dwarfing, haemorrhages, oedema and gelatinous lesions at harvest. The enzyme-linked immunosorbent assay (ELISA) results revealed a high prevalence of antibodies against CAstV amongst the broiler breeder flocks tested. It is the first study that describes the detection and prevalence of CAstV in broiler chickens and broiler breeder flocks in Malaysia.

• Adzhar, A., Gough, R. E., Haydon, D., Shaw, K., Britton, P., & Cavanagh, D. (1997). Molecular analysis of the 793/B serotype of infectious bronchitis virus in Great Britain. Avian Pathology, 26(3), 625-640. https://doi.org/10.1080/03079459708419239

• Baxendale, W., & Mebatsion, T. (2004). The isolation and characterisation of astroviruses from chickens. Avian Pathology, 33(3), 364-370. https://doi.org/10.1080/0307945042000220426

• Bosch, A., Guix, S., Krishna, N. K., Méndez, E., Monroe, S. S., Pantin-Jackwood, M., & Schultz-Cherry, S. (2012). Family-Astroviridae. In A. M. Q. King, E. Lefkowitz, M. J. Adams, & E. B. Carstens (Eds.), Ninth Report of the International Committee on Taxonomy of Viruses (pp. 953-959). Elsevier.

• Bulbule, N. R., Mandakhalikar, K. D., Kapgate, S. S., Deshmukh, V. V., Schat, K. A., & Chawak, M. M. (2013). Role of chicken astrovirus as a causative agent of gout in commercial broilers in India. Avian Pathology, 42(5), 464-473. https://doi.org/10.1080/03079457.2013.828194

• Chamings, A. (2016). Molecular epidemiology of avian nephritis virus in commercial chicken flocks (Doctoral dissertation). The University of Melbourne, Australia.

• Day, J. M., Spackman, E., & Pantin-Jackwood, M. (2007). A multiplex RT-PCR test for the differential identification of turkey astrovirus type 1, turkey astrovirus type 2, chicken astrovirus, avian nephritis virus, and avian rotavirus. Avian Diseases, 51(3), 681-684. https://doi.org/10.1637/0005-2086(2007)51[681:AMRTFT]2.0.CO;2

• Day, J. M., & Zsak, L. (2013). Recent progress in the characterization of avian enteric viruses. Avian Diseases, 57(3), 573-580. https://doi.org/10.1637/10390-092712-review.1

• De Benedictis, P., Schultz-Cherry, S., Burnham, A., & Cattoli, G. (2011). Astrovirus infections in humans and animals - Molecular biology, genetic diversity, and interspecies transmissions. Infection, Genetics and Evolution, 11(7), 1529-1544. https://doi.org/10.1016/j.meegid.2011.07.024

• de Wit, J. J., ten Dam, G. B., vande Laar, J. M. A. M., Biermann, Y., Verstegen, I., Edens, F., & Schrier, C. C. (2011). Detection and characterization of a new astrovirus in chicken and turkeys with enteric and locomotion disorders. Avian Pathology, 40(5), 453-461. https://doi.org/10.1080/03079457.2011.596813

• Erica, S., & Stephens, C. (2016). Virus isolation and propagation in embryonating eggs. In S. M. Williams, L. Dufour-Zavala, M. W. Jackwood, M. D. Lee, B. Lupiani, W. M. Reed, E. Spackman, & Peter R. Woolcock. (Eds.), A Laboratory Manual for the Isolation, Identification, and Characterization of Avian Pathogens (sixth, pp. 361-368). OmniPress Inc.

• Imada, T., Yamaguchi, S., Mase, M., Tsukamoto, K., Kubo, M., & Morooka, A. (2000). Avian nephritis virus (anv) as a new member of the family astroviridae and construction of infectious ANV cDNA. Journal of Virology, 74(18), 8487-8493. https://doi.org/10.1128/jvi.74.18.8487-8493.2000

• Kang, K. I., Linnemann, E., Icard, A. H., Durairaj, V., Mundt, E., & Sellers, H. S. (2018). Chicken astrovirus as an aetiological agent of runting-stunting syndrome in broiler chickens. Journal of General Virology, 99(4), 512-524. https://doi.org/10.1099/jgv.0.001025

• Kang, K. I., El-Gazzar, M., Sellers, H. S., Dorea, F., Williams, S. M., Kim, T., Collett, S., & Mundt, E. (2012). Investigation into the aetiology of runting and stunting syndrome in chickens. Avian Pathology, 41,1, 41-50. https://doi.org/10.1080/03079457.2011.632402

• Koci, M. D., & Schultz-Cherry, S. (2002). Avian astroviruses. Avian Pathology, 9781461447(2002), 213-227. https://doi.org/10.1007/978-1-4614-4735-1_9

• Kumar, S., Stecher, G., Li, M., Knyaz, C., & Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution, 35(6), 1547-1549. https://doi.org/10.1093/molbev/msy096

• Meulemans, G., Boschmans, M., Van den Berg, T. P., & Decaesstecker, M. (2001). Polymerase chain reaction combined with restriction enzyme analysis for detection and differentiation of fowl adenoviruses. Avian Pathology, 30(6), 655-660. https://doi.org/10.1080/03079450120092143

• Nuñez, L. F. N., Parra, S. H., Mettifogo, E., Catroxo, M. H., Astolfi-Ferreira, C. S., & Piantino Ferreira, A. J. (2015). Isolation of chicken astrovirus from specific pathogen-free chicken embryonated eggs. Poultry Science, 94(5), 947-954. https://doi.org/http://dx.doi.org/10.3382/ps/pev086

• Nuñez, L. F. N., Santander-Parra, S. H., Kyriakidis, N. C., Astolfi-Ferreira, C. S., Buim, M. R., De la Torre, D., & Ferreira, A. J. P. (2020). Molecular characterization and determination of relative cytokine expression in naturally infected day-old chicks with chicken astrovirus associated to white chick syndrome. Animals, 10(7), 1-18. https://doi.org/10.3390/ani10071195

• Palomino-Tapia, V., Mitevski, D., Inglis, T., van der Meer, F., Martin, E., Brash, M., Provost, C., Gagnon, C. A., & Abdul-Careem, M. F. (2020). Chicken astrovirus (CAstV) molecular studies reveal evidence of multiple past recombination events in sequences originated from clinical samples of white chick syndrome (WCS) in Western Canada. Viruses, 12(10), Article 1096. https://doi.org/10.3390/v12101096

• Pang, Y., Wang, H., Girshick, T., Xie, Z., & Khan, M. I. (2002). Development and application of a multiplex polymerase chain reaction for avian respiratory agents. Avian Diseases, 46(3), 691-699. https://doi.org/10.1637/0005-2086(2002)046[0691:DAAOAM]2.0.CO;2

• Pantin-Jackwood, M. J., Day, J. M., Jackwood, M. W., & Spackman, E. (2008). Enteric viruses detected by molecular methods in commercial chicken and turkey flocks in the United States between 2005 and 2006. Avian Diseases, 52(2), 235-244. https://doi.org/10.1637/8174-111507-Reg.1

• Patel, A. K., Pandit, R. J., Thakkar, J. R., Hinsu, A. T., Pandey, V. C., Pal, J. K., Prajapati, K. S., Jakhesara, S. J., & Joshi, C. G. (2017). Complete genome sequence analysis of chicken astrovirus isolate from India. Veterinary Research Communications, 41(1), 67-75. https://doi.org/10.1007/s11259-016-9673-6

• Pérot, P., Lecuit, M., & Eloit, M. (2017). Astrovirus diagnostics. Viruses, 9(1), 1-14. https://doi.org/10.3390/v9010010

• Sajewicz-Krukowska, J., Pać, K., Lisowska, A., Pikuła, A., Minta, Z., Króliczewska, B., & Domańska-Blicharz, K. (2016). Astrovirus-induced “white chicks” condition - Field observation, virus detection and preliminary characterization. Avian Pathology, 45(1), 2-12. https://doi.org/10.1080/03079457.2015.1114173

• Shah, J. D., Desai, P. T., Zhang, Y., Scharber, S. K., Baller, J., Xing, Z. S., & Cardona, C. J. (2016). Development of the intestinal RNA virus community of healthy broiler chickens. PLoS ONE, 11(2), 1-13. https://doi.org/10.1371/journal.pone.0150094

• Skibinska, A., Lee, A., Wylie, M., Smyth, V. J., Welsh, M. D., & Todd, D. (2015). Development of an indirect enzyme-linked immunosorbent assay test for detecting antibodies to chicken astrovirus in chicken sera. Avian Pathology, 44(6), 436-442. https://doi.org/10.1080/03079457.2015.1084411

• Smyth, V. J. (2017). A review of the strain diversity and pathogenesis of chicken astrovirus. Viruses, 9(2), 1-10. https://doi.org/10.3390/v9020029

• Smyth, V. J., Jewhurst, H. L., Adair, B. M., & Todd, D. (2009). Detection of chicken astrovirus by reverse transcriptase-polymerase chain reaction. Avian Pathology, 38(4), 293-299. https://doi.org/10.1080/03079450903055397

• Smyth, V., Trudgett, J., Wylie, M., Jewhurst, H., Conway, B., Welsh, M., Kaukonen, E., & Perko-Mäkelä, P. (2013). Chicken astrovirus detected in hatchability problems associated with ‘white chicks.’ Veterinary Record, 173(16), 403-404. https://doi.org/10.1136/vr.f6393

• Tang, Y., Ismail, M. M., & Saif, Y. M. (2006). Development of antigen-capture enzyme-linked immunosorbent assay and RT-PCR for detection of turkey astroviruses. Avian Diseases, 49(2), 182-188. https://doi.org/10.1637/7255-080504r

• Todd, D., Smyth, V. J., Ball, N. W., Donnelly, B. M., Wylie, M., Knowles, N. J., & Adair, B. M. (2009a). Identification of chicken enterovirus-like viruses, duck hepatitis virus type 2 and duck hepatitis virus type 3 as astroviruses. Avian Pathology, 38(1), 21-29. https://doi.org/10.1080/03079450802632056

• Todd, D., Wilkinson, D. S., Jewhurst, H. L., Wylie, M., Gordon, A. W., & Adair, B. M. (2009b). A seroprevalence investigation of chicken astrovirus infections. Avian Pathology, 38(4), 301-309. https://doi.org/10.1080/03079450903055421

• Xue, J., Han, T., Xu, M., Zhao, J., & Zhang, G. (2017). The first serological investigation of Chicken astrovirus infection in China. Biologicals, 47, 22-24. https://doi.org/10.1016/j.biologicals.2017.03.005

• Xue, J., Han, T., Zhao, Y., Yang, H., & Zhang, G. (2020). Complete genome sequence and phylogenetic analysis of novel avastroviruses circulating in China from 2016 to 2018. Virus Research, 278, Article 197858. https://doi.org/10.1016/j.virusres.2020.197858

• Zsak, L., Cha, R. M., & Day, J. M. (2013). Chicken parvovirus-induced runting-stunting syndrome in young broilers. Avian Diseases, 57(1), 123-127. https://doi.org/10.1637/10371-091212-ResNote.1