The role of the major viral pathogens in a respiratory disease outbreak of broiler flocks in Eastern Iran

Document Type : Full paper (Original article)


1 Department of Biotechnology and Veterinary Research, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Mashhad, Iran

2 Ph.D. Student in Medical Virology, Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences (current address); Doctorate in Veterinary Medicine, South Khorasan Veterinary Service Head Office, Birjand, Iran

3 Doctorate in Veterinary Medicine, South Khorasan Veterinary Service Head Office, Birjand, Iran


Background: Infectious bronchitis virus (IBV), Newcastle disease virus (NDV), and avian influenza virus (AIV) H9N2 are major viral pathogens in broiler respiratory disease. Aims: Following a respiratory disease outbreak and economic losses in eastern Iran 2020-2021, we investigated the role of major viral pathogens and the implemented vaccination programs. Methods: Thirty-six respiratory disease affected broiler flocks in South Khorasan province were sampled, molecularly tested, and coinfections were investigated. The vaccination programs were obtained and the detected IBV were genotyped. Results: IBV, virulent NDV, and AIV H9N2 were detected in twenty-five, seven, and seven flocks, respectively. IBV+AIV, IBV+NDV, and NDV+AIV coinfections were respectively detected in six, five, and one flocks. Most IBV infected flocks (84%) had been immunized with a live IBV-Mass vaccine. All NDV infected flocks and 14.2% of AIV infected flocks had been vaccinated. IBV genotyping showed a high prevalence of variant 2 (83.3%), followed by Mass-type (12.5%), and Q1-type (4.2%). Variant 2 IB viruses were widely distributed in the province and half of them were mostly similar to the ones that had been detected in northern neighboring province, Khorasan Razavi. Conclusion: Single infection with variant 2 IBV was a major cause of the respiratory disease outbreak in which use of the Mass vaccine was probably not effective. The high coverage and multiple doses of vaccination against Newcastle disease possibly had reduced the prevalence of NDV. Considering the regional origin of IBV strains, strong biosecurity measures should be implemented and vaccination programs using appropriate vaccine strains should be used.


Main Subjects

Al-Ali, AM; El-Sabagh, IM; Mohamed, MH; Alluwaimi, AM and Arif, IA (2018). Molecular characterization of common respiratory viral infections in broilers in Al-Hassa, Eastern Province, Saudi Arabia. Thai. J. Vet. Med., 48: 235-245.
Alian Samakkhah, S; Bahonar, A; Zaynolabedini Tehrani, F; Ghafouri, SA; Sadrzadeh, A and Fallah Mehrabadi, MH (2019). Occurrence of Newcastle disease in Iranian broiler farms during 2013-2015. J. Vet. Res., 74: 1-10.
Allahyari, E; Allymehr, M; Molouki, A and Fallah, M (2022). Molecular characterisation and phylogenetic study of the fusion gene of Newcastle disease viruses isolated from broiler farms of Iran in 2018-2019. Bulg. J. Vet. Med., 25: 21-32.
Awad, F; Forrester, A; Baylis, M; Lemiere, S; Ganapathy, K; Hussien, HA and Capua, I (2015). Protection conferred by live infectious bronchitis vaccine viruses against variant Middle East IS/885/00-like and IS/1494/06-like isolates in commercial broiler chicks. Vet. Rec. Open. 2: e000111.
Cavanagh, D (1983). Coronavirus IBV glycopolypeptides: size of their polypeptide moieties and nature of their oligosaccharides. J. Gen. Virol., 64: 1187-1191.
Cavanagh, D; Davis, PJ and Mockett, AA (1988). Amino acids within hypervariable region 1 of avian coronavirus IBV (Massachusetts serotype) spike glycoprotein are associated with neutralization epitopes. Virus Res., 11: 141-150.
Cavanagh, D; Mawditt, K; Britton, P and Naylor, C (1999). Longitudinal field studies of infectious bronchitis virus and avian pneumovirus in broilers using type-specific polymerase chain reactions. Avian Pathol., 28: 593-605.
De Wit, J; Dijkman, R; Guerrero, P; Calvo, J; Gonzalez, A and Hidalgo, H (2017). Variability in biological behaviour, pathogenicity, protectotype and induction of virus neutralizing antibodies by different vaccination programmes to infectious bronchitis virus genotype Q1 strains from Chile. Avian Pathol., 46: 666-675.
De Wit, J; Guerrero, P; Calvo, J and Hidalgo, H (2012). Report of the genotyping, pathotyping, and protectotyping of recent strains from Chile. Paper presented at the 7th International Symposium on Avian Corona- and Pneumo-viruses and Complicating Pathogens. Rauischholzhausen, Germany. 18-21 June 2012.
Dimitrov, KM; Abolnik, C; Afonso, CL; Albina, E; Bahl, J; Berg, M; Briand, FX; Brown, IH; Choi, KS; Chvala, I; Diel, DG; Durr, PA; Ferreira, HL; Fusaro, A; Gil, P; Goujgoulova, GV; Grund, C; Hicks, JT; Joannis, TM; Torchetti, MK; Kolosov, S; Lambrecht, B; Lewis, NS; Liu, H; Liu, H; McCullough, S; Miller, PJ; Monne, I; Muller, CP; Munir, M; Reischak, D; Sabra, M; Samal, SK; Servan de Almeida, R; Shittu, I; Snoeck, CJ; Suarez, DL; Van Borm, S; Wang, Z and Wong, FYK (2019). Updated unified phylogenetic classification system and revised nomenclature for Newcastle disease virus. [J]. Infect. Genet. Evol., 74: 103917. doi: 10.1016/j.meegid. 2019.103917.
Dimitrov, KM; Afonso, CL; Yu, Q and Miller, PJ (2017). Newcastle disease vaccines—A solved problem or a continuous challenge?. Vet. Microbiol., 206: 126-136.
Ebrahimi, M; Grigorian, S; Fallah Mehrabadi, M and Shoushtari, A (2021). Serological survey of H9N2 influenza viruses in rural chicken of Northern provinces, Iran. Vet. Res. Biol. Pro., 34: 15-25.
Elbestawy, A; Ellakany, H; Sedeik, M; Gado, A; Abdel-Latif, M; Noreldin, A; Orabi, A; Radwan, I and El-Ghany, WA (2023). Superior efficacy of apathogenic genotype I (V4) over lentogenic genotype II (LaSota) live vaccines against Newcastle disease virus genotype VII.1.1 in pathogen-associated molecular pattern-H9N2 vaccinated broiler chickens. Vaccines. 11: 1-19.
Fallah Mehrabadi, MH; Motamed, N; Ghalyanchilangeroudi, A and Tehrani, F (2020). Avian influenza (H9N2 subtype) in Iranian broiler farms: A cross-sectional study. Arch. Razi Inst., 75: 359-366.
Fawzy, M; Ali, RR; Elfeil, WK; Saleh, AA and El-Tarabilli, MMA (2020). Efficacy of inactivated velogenic Newcastle disease virus genotype VII vaccine in broiler chickens. Vet. Res. Forum. 11: 113-120.
Ghalyanchilangeroudi, A; Hosseini, H; Mehrabadi, MHF; Ghafouri, SA; Hamdan, AM; Ziafati, Z; Dizaji, RE and Mohammadi, P (2019). Genotyping of avian infectious bronchitis virus in Iran: Detection of D274 and changing in the genotypes rate. Comp. Immunol. Microbiol. Infect. Dis., 65: 110-115.
Ghalyanchi-Langeroudi, A; Karimi, V; Jannat, A; Hashemzadeh, M; Fallah, M; Gholami, F; Zabihi, M and Heidarzadeh, M (2015). Genotyping of infectious bronchitis viruses in the East of Iran, 2015. Iran. J. Virol., 9: 31-35.
Ghalyanchilangeroudi, A; Najafi, H; Mehrabadi, MF; Kafi, ZZ; Sadri, N; Rajeoni, AH; Modiri, A; Safari, A and Hosseini, H (2020). The emergence of Q1 genotype of avian infectious bronchitis virus in Iran, 2019: the first report. Iran. J. Vet. Res., 21: 230-233.
Gholami, F; Karimi, V; Ghalyanchi Langeroudi, A; Hashemzadeh, M and Vasfi Marandi, M (2018). Genotyping of infectious bronchitis viruses isolated from broiler chicken farms in Iran during 2015-2016. Iran. J. Vet. Med., 12: 9-17.
Ghorbiani, M; Boroomand, Z; Mayahi, M and Seyfi Abad Shapouri, MR (2020). Molecular identification of infectious bronchitis virus isolated from respiratory diseases in some Iranian broiler flocks. Mol. Biol. Rep., 47: 7161-7168.
Haji-Abdolvahab, H; Ghalyanchilangeroudi, A; Bahonar, A; Ghafouri, SA; Vasfi Marandi, M; Mehrabadi, MHF and Tehrani, F (2019). Prevalence of avian influenza, Newcastle disease, and infectious bronchitis viruses in broiler flocks infected with multifactorial respiratory diseases in Iran, 2015-2016. Trop. Anim. Health Prod., 51: 689-695.
Hall, T; Biosciences, I and Carlsbad, C (2011). BioEdit: an important software for molecular biology. GERF Bull. Biosci., 2: 60-61.
Hamadan, AM; Ghalyanchilangeroudi, A; Hashemzadeh, M; Hosseini, H; Karimi, V; Yahyaraeyat, R and Najafi, H (2017). Genotyping of avian infectious bronchitis viruses in Iran (2015-2017) reveals domination of IS-1494 like virus. Virus Res., 240: 101-106.
Hejazi, Z; Tabatabaeizadeh, SE; Toroghi, R; Farzin, H and Saffarian, P (2022). First detection and characterisation of sub-genotype XIII. 2.1 Newcastle disease virus isolated from backyard chickens in Iran. Vet. Med. Sci., 8: 2521-2531.
Homayounimehr, A; Pakbin, A; Momayyez, R and Fatemi, SMR (2016). Detection and identification of infectious bronchitis virus by RT-PCR in Iran. Trop. Anim. Health Prod., 48: 973-978.
Hosseini, H; Fard, MHB; Charkhkar, S and Morshed, R (2015). Epidemiology of avian infectious bronchitis virus genotypes in Iran (2010-2014). Avian Dis., 59: 431-435.
Hosseini, H; Ghalyanchilangeroudi, A; Fallah Mehrabadi, MH; Sediqian, MS; Shayeganmehr, A; Ghafouri, SA; Maghsoudloo, H; Abdollahi, H and Farahani, RK (2017). Phylogenetic analysis of H9N2 avian influenza viruses in Afghanistan (2016-2017). Arch. Virol., 162: 3161-3165.
ICTV (2018). Virus Taxonomy. Retrieved from https://talk.
Kant, A; Koch, G; Van Roozelaar, D; Balk, F and Huurne, AT (1997). Differentiation of virulent and non-virulent strains of Newcastle disease virus within 24 hours by polymerase chain reaction. Avian Pathol., 26: 837-849.
Khabiri, A; Toroghi, R; Mohammadabadi, M and Tabatabaeizadeh, SE (2023). Introduction of a Newcastle disease virus challenge strain (sub-genotype VII.1.1) isolated in Iran. Vet. Res. Forum. 14: 221-228.
Kumar, S; Stecher, G; Li, M; Knyaz, C and Tamura, K (2018). MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol., 35: 1547-1549.
Lee, MS; Chang, PC; Shien, JH; Cheng, MC and Shieh, HK (2001). Identification and subtyping of avian influenza viruses by reverse transcription-PCR. J. Virol. Methods. 97: 13-22.
Lupiani, B and Reddy, SM (2009). The history of avian influenza. Comp. Immunol. Microbiol. Infect. Dis., 32: 311-323.
Maki, F; Boroomand, Z; Mayahi, M and Seyfi Abad Shapouri, M (2022). The role of Newcastle disease virus in respiratory syndromes of broiler flocks in Khuzestan and Isfahan provinces, Iran. Vet. Res. Biol. Pro., 35: 72-81.
Malik, YS; Patnayak, DP and Goyal, SM (2004). Detection of three avian respiratory viruses by single-tube multiplex reverse transcription-polymerase chain reaction assay. J. Vet. Diagn., 16: 244-248.
Meir, R; Rosenblut, E; Perl, S; Kass, N; Ayali, G; Hemsani, E and Perk, S (2004). Identification of a novel nephropathogenic infectious bronchitis virus in Israel. Avian Dis., 48: 635-641.
Miller, PJ; Estevez, C; Yu, Q; Suarez, DL and King, DJ (2009). Comparison of viral shedding following vaccination with inactivated and live Newcastle disease vaccines formulated with wild-type and recombinant viruses. Avian Dis., 53: 39-49.
Miller, PJ; King, DJ; Afonso, CL and Suarez, DL (2007). Antigenic differences among Newcastle disease virus strains of different genotypes used in vaccine formulation affect viral shedding after a virulent challenge. Vaccine. 25: 7238-7246.
Molouki, A; Mehrabadi, MHF; Bashashati, M; Akhijahani, MM; Lim, SHE and Hajloo, SA (2019). NDV subgenotype VII (L) is currently circulating in commercial broiler farms of Iran, 2017-2018. Trop. Anim. Health Prod., 51: 1247-1252.
Molouki, A; Sotani, M; Fallah Mehrabadi, MH; Shoushtari, A; Abtin, A; Mahmoudzadeh Akhijahani, M; Abdoshah, M; Pourbakhsh, SA; Allahyari, E and Ghalyanchilangeroudi, A (2021). Predominance of fourth panzootic Newcastle disease virus subgenotype VII. 1.1 in Iran and its relation to the genotypes circulating in the region. Curr. Microbiol., 78: 3068-3078.
Naeem, K; Siddique, N; Ayaz, M and Jalalee, M (2007). Avian influenza in Pakistan: outbreaks of low- and high-pathogenicity avian influenza in Pakistan during 2003-2006. Avian Dis., 51: 189-193.
Nili, H and Asasi, K (2003). Avian influenza (H9N2) outbreak in Iran. Avian Dis., 47: 828-831.
Pantin-Jackwood, MJ and Spackman, E (2020). Multicausal respiratory diseases. In: Swayne, DE; Boulianne, M; Logue, CM; McDougald, LR; Nair, V and Suarez, DL (Eds.), Diseases of poultry (14th Edn.), Ames, Iowa, Iowa State University Press. PP: 1386-1390.
Rahimi, A; Zamani-moghadam, AK and Shoshtari, AH (2018). Phylogenetic and genotypic study of infectious bronchitis virus isolated from broiler herds by RT-PCR and S1 gene sequence analysis. Iran. J. Vet. Clin. Sci., 12: 81-87.
Roohani, K; Tan, SW; Yeap, SK; Ideris, A; Bejo, MH and Omar, AR (2015). Characterisation of genotype VII Newcastle disease virus (NDV) isolated from NDV vaccinated chickens, and the efficacy of LaSota and recombinant genotype VII vaccines against challenge with velogenic NDV. J. Vet. Sci., 16: 447-457.
Roussan, D; Haddad, R and Khawaldeh, G (2008). Molecular survey of avian respiratory pathogens in commercial broiler chicken flocks with respiratory diseases in Jordan. Poult. Sci., 87: 444-448.
Schalk, AF and Hawn, MC (1931). An apparently new respiratory disease of baby chicks. J. Am. Vet. Med. Assoc., 78: 413-422.
Sid, H; Benachour, K and Rautenschlein, S (2015). Co-infection with multiple respiratory pathogens contributes to increased mortality rates in Algerian poultry flocks. Avian Dis., 59: 440-446.
Suarez, DL; Miller, PJ; Koch, G; Mundt, E and Rautenschlein, S (2020). Newcastle disease, other avian
paramyxoviruses, and avian metapneumovirus infections. In: Swayne, DE; Boulianne, M; Logue, CM; McDougald, LR; Nair, V; Suarez, DL; De Wit, S; Grimes, T; Johnson, D; Kromm, M; Prajitno, TY; Rubinoff, I and Zavala, G (Eds.), Diseases of poultry. (14th Edn.), Ames, Iowa, Iowa State University Press. PP: 109-166.
Tabatabaeizadeh, SE (2021). Immunoinformatics analysis and antibody epitope comparison of Newcastle disease virus classical vaccines with a virus involved in the fifth NDV panzootic. Res. Sq. [Preprint]. Available from:
Tabatabaeizadeh, SE; Toroghi, R; Margan Azghadi, N; Farzin, H; Sharghi, S; Sarani, M; Jamshidian Mojaver, M; Alami Abardeh, J; Ghorbanzadeh, M and Torabi, M (2021). Detection of mutant infectious bronchitis viruses of GI-23 lineage from commercial chicken flocks in Khorasan Razavi province, Iran in 2019. Iran. J. Vet. Sci. Technol., 13: 14-21.
Toffan, A; Bonci, M; Bano, L; Valastro, V; Vascellari, M; Capua, I and Terregino, C (2013). Diagnostic and clinical observation on the infectious bronchitis virus strain Q1 in Italy. Vet. Ital., 49: 347-355.
Umar, S; Teillaud, A; Aslam, HB; Guerin, JL and Ducatez, MF (2019). Molecular epidemiology of respiratory viruses in commercial chicken flocks in Pakistan from 2014 through to 2016. BMC Vet. Res., 15: 1-12.
Valastro, V; Holmes, EC; Britton, P; Fusaro, A; Jackwood, MW; Cattoli, G and Monne, I (2016). S1 gene-based phylogeny of infectious bronchitis virus: an attempt to harmonize virus classification. Infect. Genet. Evol., 39: 349-364.
Worthington, KJ; Currie, R and Jones, RC (2008). A reverse transcriptase-polymerase chain reaction survey of infectious bronchitis virus genotypes in Western Europe from 2002 to 2006. Avian Pathol., 37: 247-257.
Yu, L; Jiang, Y; Low, S; Wang, Z; Nam, SJ; Liu, W and Kwang, J (2001). Characterization of three infectious bronchitis virus isolates from China associated with proventriculus in vaccinated chickens. Avian Dis., 45: 416-424.