Seroprevalence of Newcastle disease virus and avian influenza virus in poultry and captive wild birds in poultry-dense regions of Pakistan


Newcastle disease virus, Avian influenza virus, seroprevalence, poultry, captive wild birds, poultry-dense regions, Pakistan

How to Cite

Aziz, U.-R., Shabbir, M. A. B. ., Iqbal, M. Z. ., Yasin, R. ., Ishaq, H. M., Mehmood, A. ., Yousaf, F. ., Rasheed, M. ., Rasul, S. ., Usman, M. ., & Raza, M. A. . (2023). Seroprevalence of Newcastle disease virus and avian influenza virus in poultry and captive wild birds in poultry-dense regions of Pakistan. Veterinaria Italiana, 59(1). https://doi.org/10.12834/VetIt.2449.17415.2


Newcastle disease virus (NDV) and avian influenza virus (AIV) are causing contagious diseases in chickens and wild birds worldwide; however, there is a paucity of information on the current status of seropositivity of Newcastle and avian influenza diseases in chickens and wild birds of Pakistan. Therefore, the present study aimed to investigate the serological evidence of both diseases in commercial poultry (broiler, layer chickens), backyard poultry, and captive wild birds in poultry‑dense regions of Punjab, Pakistan. Enzyme‑linked immunosorbent (ELISA) and haemagglutination inhibition (HI) assays were performed for the determination of antibodies against NDV and AIV and their genotyping and subtyping, respectively. Overall, 47.5% and 67.4% seroprevalence of NDV and AIV, respectively, was observed in both poultry and wild birds. Based on bird’s category, layer chickens had the highest seroprevalence of NDV (60.8%, 95% CI: 52.95‑68.22, OR: 0.71) followed by backyard poultry (56.8%, 95% CI: 47.92‑65.32, OR: 0.82), broilers (52.7%, 95% CI: 46.84‑58.64), pigeons (41.3%, 95% CI: 30.53‑52.81, OR: 1.59), peafowls (26.1%, 95% CI: 11.09‑48.69, OR: 3.16), ducks (23.8%, 95% CI: 12.59‑39.8, OR: 3.57), turkeys (16.7%, 95% CI: 4.41‑42.27, OR: 5.58), parrots (14.3%, 95% CI: 2.52‑43.85, OR: 6.70) and quails (2.3%, 95% CI: 0.2‑13.51, OR: 4.8). Comparatively, backyard chickens had the highest seroprevalence of AIV (78.8%, 95% CI: 70.64‑85.22, OR: 0.63) followed by ducks (73.8%, 95% CI: 57.68‑85.6, OR: 0.83), layers (73.5%, 95% CI: 65.98‑79.89, OR: 0.84), pigeons (72.5%, 95% CI: 61.2‑81.61, OR: 0.89), broilers (70.1%, 95% CI: 64.44‑75.29), turkeys (55.5%, 95% CI: 31.35‑77.6, OR: 1.87), peafowls (47.8%, 95% CI: 27.42‑68.9, OR: 2.56) and parrots (42.8%, 95% CI: 18.8‑70.3, OR: 3.1). Overall, 40.1%, 34.2%, 31.3%, and 25.1% sera were positive for H9 AIV, G‑VII NDV, H7 AIV, and G‑VI NDV, respectively. The current study revealed a widespread exposure to NDV and AIV in poultry and captive wild birds. Therefore, it is crucial to include captive wild birds in NDV and AIV surveillance programs to further strengthen disease control measures, particularly in endemic regions.



Abdelwhab, E.M., Veits, J., Ulrich, R., Kasbohm, E., Teifke, J.P. & Mettenleiter, T.C. 2016. Composition of the hemagglutinin polybasic proteolytic cleavage motif mediates variable virulence of H7N7 avian influenza viruses. Scientific Reports, 6, 39505.
Aiki-Raji, C.O., Adebiyi, A.I., Agbajelola, V.I., Adetunji, S.A., Lameed, Q., Adesina, M., Adekanye, G., Omidokun, F., Fagbohun, O. & Oluwayelu, D.O. 2015. Surveillance for low pathogenic avian influenza viruses in live-bird markets in Oyo and Ogun States, Nigeria. Asian Pacific J Tropical Dis, 5, 369-373.
Al Shekaili, T., Clough, H., Ganapathy, K. & Baylis, M. 2015. Serosurveillance and risk factors for avian influenza and Newcastle disease virus in backyard poultry in Oman. Prev Vet Med, 122(1–2), 145–153. https://doi.org/10.1016/j.prevetmed.2015.09.011
Amarasinghe, G.K., Ayllón, M.A., Bào, Y., Basler, C.F., Bavari, S., Blasdell, K.R., Briese, T., Brown, P.A., Bukreyev, A., Balkema-Buschmann, A. & Buchholz, U.J. 2019. Taxonomy of the order Mononegavirales: update 2019. Arch Virol, 164, 1967–1980.
Aziz-ul-Rahman., Habib, M., Riaz, T., Hussain, B., Yousaf, F., Saqalein, M. & Rasool, M.H. 2017. Seroprevalence of newcastle disease virus (NDV) in commercial and domesticated birds: Pakistan during current surge of NDV. J Infect Mol Biol, 4(4), 54-59.
Aziz-ul-Rahman., Rohaim, M.A., El Naggar, R.F., Mustafa, G., Chaudhry, U. & Shabbir, M.Z. 2019a. Comparative clinico-pathological assessment of velogenic (sub-genotype VIIi) and mesogenic (sub-genotype VIm) Avian avulavirus 1 in chickens and pigeons. Avian Pathol, 48(6), 610-621.
Aziz-ul-Rahman., Yaqub, T., Imran, M., Habib, M., Sohail, T., Mukhtar, N., Shahid, M.F., Munir, M. & Shabbir, M.Z. 2019b. Sequence analysis and biological characterization of virulent avian avulavirus 1 isolated from asymptomatic migratory fowl. Acta Virol, 63, 223-228.
Aziz-ul-Rahman., Yaqub, T., Imran, M., Habib, M., Sohail, T., Furqan Shahid, M., Munir, M. & Shabbir, M.Z. 2018. Phylogenomics and infectious potential of Avian Avulaviruses species-type 1 isolated from healthy green-winged teal (Anas carolinensis) from a wetland sanctuary of indus river. Avian Dis, 62(4), 404-415.
Bertran, K., Dolz, R. & Majó, N. 2014. Pathobiology of avian influenza virus infection in minor gallinaceous species: a review. Avian Pathol, 43(1), 9-25.
Biswas, P.K., Barua, H., Uddin, G.M.N., Biswas, D., Ahad, A. & Debnath, N.C. 2009. Serosurvey of five viruses in chickens on smallholdings in Bangladesh. Prev Vet Med, 88, 67-71.
Brown, V.R. a7 Bevins, S.N. 2017. A review of virulent Newcastle disease viruses in the United States and the role of wild birds in viral persistence and spread. Vet Res, 48(1), 1-15.
Caron, A., Cappelle, J., Cumming, G.S., de Garine-Wichatitsky, M. & Gaidet, N. 2015. Bridge hosts, a missing link for disease ecology in multi-host systems. Vet Res, 46, 83.
Chatziprodromidou, I.P., Arvanitidou, M., Guitian, J., Apostolou, T., Vantarakis, G. & Vantarakis, A. 2018. Global avian influenza outbreaks 2010–2016: a systematic review of their distribution, avian species and virus subtype. Systematic Reviews, 7(1), 1-12.
Couacy-Hymann, E., Kouakou, A.V., Kouame, C.K., Kouassi, A.L., Koffi, Y.M., Godji, P., et al., 2012. Surveillance for avian influenza and Newcastle disease in backyard poultry flocks in Cote d’Ivoire, 2007–2009. Rev Sci Tech, 31, 821–8.
Dimitrov, K.M., Ramey, A.M., Qiu, X., Bahl, J. & Afonso, C.L. 2016. Temporal, geographic, and host distribution of avian paramyxovirus 1 (Newcastle disease virus). Infect Genet Evol, 39, 22-34.
East, I., Kite, V., Daniels, P. & Garner, G. 2006. A cross-sectional survey of Australian chicken farms to identify risk factors associated with seropositivity to Newcastle-disease virus. Prev Vet Med, 77(3-4), 199-214.
Fatima, Z., Khan, M.A., Ahmad, M.U.D., Muhammad, K., Khwaja, K.N., Khan, A., Anwar, Z., Ahad, A. & Mahmood, A. 2017. Cross sectional survey of live bird markets, and zoo birds for circulating influenza subtypes in Pakistan. Pakistan Vet J, 37, 185-189.
Gutierrez-Ruiz, E.J., Ramirez-Cruz, G.T., Camara Gamboa, E.I., Alexander, D.J. & Gough, R.E.A. 2000. Serological survey for avian infectious bronchitis virus and Newcastle disease virus antibodies in backyard (free-range) village chickens in Mexico. Trop Anim Health Prod, 32, 381-390.
Hassan, M.Z., Rahman, M.M., Das, B.C., Al Amin, M., Sultana, S., Ferdouse, H., Jaber, M., Rahman, M.S. & Hoque, M.F. 2018. Epidemiology of duck as reservoir of Avian Influenza Virus in Bangladesh. Asian J Med Biol Res, 4(1), 14-20.
Hua, Y.P., Chai, H.L., Yang, S.Y., Zeng, X.W. & Sun, Y. 2005. Primary survey of avian influenza virus and Newcastle disease virus infection in wild birds in some areas of Heilongjiang Province, China. J Vet Sci, 6(4).
Hurtado, R. & Vanstreels, R.E.T. 2016. Avian influenza in wild birds from South America: Review, implications and perspectives. Exploratory Res Hypothesis Med, 1(4), 62-74.
Kausar, A., Anwar, S., Siddique, N., Ahmed, S. & Dasti, J.I. 2018. Prevalence of avian influenza H9N2 virus among wild and domesticated bird species across Pakistan. Pakistan J Zool, 50(4), 1347-1354.
Lee, E.K., Kang, H.M., Song, B.M., Lee, Y.N., Heo, G.B., Lee, H.S., Lee, L.J. & Kim, J.H. 2017. Surveillance of avian influenza viruses in South Korea between 2012 and 2014. Virol J, 14, 54. https://doi.org/10.1186/s12985-017-0711-y
McQuiston, J.H., Garber, L.P., Porter-Spalding, B.A., Hahn, J.W., Pierson, F.W., Wainwright, S.H., Senne, D.A., Brignole, T.J., Akey, B.L. & Holt, T.J. 2005. Evaluation of risk factors for the spread of low pathogenicity H7N2 avian influenza virus among commercial poultry farms. J Amer Vet Med Assoc, 226(5), 767-772.
Miller, P.J., Haddas, R., Simanov, L., Lublin, A., Rehmani, S.F., Wajid, A., Bibi, T., Khan, T.A., Yaqub, T., Setiyaningsih, S. & Afonso, C.L. 2015. Identification of new sub-genotypes of virulent Newcastle disease virus with potential panzootic features. Infect Genet Evol, 29, 216-229.
Munir, M., Cortey, M., Abbas, M., Afzal, F., Shabbir, M.Z., Khan, M.T., Ahmed, S., Ahmad, S., Baule, C., Ståhl, K. & Zohari, S. 2012. Biological characterization and phylogenetic analysis of a novel genetic group of Newcastle disease virus isolated from outbreaks in commercial poultry and from backyard poultry flocks in Pakistan. Infect Genet Evol, 12(5), 1010-1019.
Munir, T., Aslam, A., Zahid, B., Ahmed, I., Imran, M.S. & Ijaz, M. 2015. Potential of commonly resident wild birds towards newcastle disease virus transmission. Pakistan Vet J, 35(1), 106-107.
Naeem, K., Siddique, N., Ayaz, M. & Jalalee, M.A. 2007. Avian influenza in Pakistan: outbreaks of low-and high-pathogenicity avian influenza in Pakistan during 2003–2006. Avian Dis, 51(s1), 189-193.
OIE. 2012. Newcastle Disease. Biological Standards Commission, Manual of Diagnostic Tests and Vaccines for Terrestrial Animals: Mammals, Birds andBees. World Organisation for Animal Health, Paris, France, 555–574.
Panda, A., Huang, Z., Elankumaran, S., Rockemann, D.D. & Samal, S.K. 2004. Role of fusion protein cleavage site in the virulence of Newcastle disease virus. Micro Pathogen, 36(1), 1-10.
Pedersen, J.C., Senne, D.A., Woolcock, P.R., Kinde, H., King, D.J., Wise, M.G., Panigrahy, B. & Seal, B.S. 2004. Phylogenetic relationships among virulentNewcastle disease virus isolates from the 2002–2003 outbreak in Californiaand other recent outbreaks in North America. J Clin Microbiol, 42, 2329–2334.
Rahman, A.U., Munir, M. & Shabbir, M.Z. 2019. A comparative genomic and evolutionary analysis of circulating strains of avian avulavirus 1 in Pakistan. Mol Genet Genom, 294(5), 1289-1309.
Saadat, Y., Ghafouri, S.A., Tehrani, F. & Langeroudi, A.G. 2014. An active serologicalsurvey of antibodies to newcastle disease and avian influenza (H9N2) virusesin the unvaccinated backyard poultry in Bushehr province, Iran, 2012–2013. Asian Pacific J Trop Biomed, 4, S213–216.
Sarwar, M., Muhammad, K., Rabbani, M., Younus, M., Sarwar, N., Ali, M.A. & Ahad, A. 2013. Prevalence of avian influenza viruses in live bird markets of Lahore. J Anim Plant Sci, 23, 388-92.
Shabbir, M.Z., Zohari, S., Yaqub, T., Nazir, J., Shabbir, M.A.B., Mukhtar, N., Shafee, M., Sajid, M., Anees, M., Abbas, M. & Khan, M.T. 2013. Genetic diversity of Newcastle disease virus in Pakistan: a countrywide perspective. Virol J, 10(1), 170.
Sonia, M. D., Hernandez, P., Villegas, F., Prieto, J. C., Unda, N., Stedman, B., Ritchie, R. & Stephen, J. 2006. A survey of selected avian pathogens of backyard poultry in northwestern Ecuador. J Avian Med Surgery, 20, 147–158.
Ssematimba, A., Hagenaars, T.J., de Wit, J.J., Ruiterkamp, F., Fabri, T.H., Stegeman, J.A. & de Jong, M.C. 2013. Avian influenza transmission risks: analysis of biosecurity measures and contact structure in Dutch poultry farming. Pre Vet Med, 109(1-2), 106-15.
Swayne, D.E. & King, D.J. 2003. Avian influenza and Newcastle disease. J Amer Vet Med Asso, 222, 1534–1540.
Tadesse, S., Ashenafi, H. & Zeleke, A. 2005. Seroprevalence study of Newcastle disease in local chickens in Central Ethiopia. Inter J Applied Res Vet Med, 3(1), 25-29.
Terregino, C., De Nardi, R., Guberti, V., Scremin, M., Raffini, E., Moreno Martin, A., Cattoli, G., Bonfanti, L. & Capua, I. 2007. Active surveillance for avian influenza viruses in wild birds and backyard flocks in Northern Italy during 2004 to 2006. Avian Pathol, 36(4), 337-344.
Tombari, W., Paul, M., Bettaieb, J., Larbi, I., Nsiri, J., Elbehi, I., Gribaa, L. & Ghram, A. 2013. Risk factors and characteristics of low pathogenic avian influenza virus isolated from commercial poultry in Tunisia. PLoS One, 8(1), e53524.
Wajid, A., Dundon, W.G., Hussain, T. & Babar, M.E. 2018. Pathotyping and genetic characterization of avian avulavirus-1 from domestic and wild waterfowl, geese and black swans in Pakistan, 2014 to 2017. Arch Virol, 163(9), 2513-2518.
Wang, Y., Jiang, Z., Jin, Z., Tan, H. & Xu, B. 2013. Risk factors for infectious diseases in backyard poultry farms in the Poyang Lake area, China. PLoS One, 8(6), e67366.
Wong, J.K., Wilcox, B.R., Fojtik, A., Poulson, R.L. & Stallknecht, D.E. 2016. Antibodies to influenza A viruses in wintering snow geese (Chen caerulescens) in Texas. Avian Dis, 60, 337-340. https://doi.org/10.1637/11104-042115-RegR
Yongolo, M.G., Maeda-machang’u, S.A.D. & Minga, U.M. 2001. Newcastle disease and infectious bursal disease among free-range village chickens in Tanzania. Proceeding Assoc. Inst. Tropic. Vet. Med. (AIMVT). Assoc. Inst. Tropic. Vet. Med. Copenhagen, Denmark.
Copyright (c) 2023 Ul-Rahman Aziz