Evaluation of viral load and expression level of apoptotic genes in selected tissues of two hybrids of commercial broiler chickens challenged with infectious bronchitis virus: a comparative study

Document Type : Full paper (Original article)


1 Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Karaj Branch, Islamic Azad University, Karaj, Iran

3 Department of Poultry Diseases, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran


Background: Infectious bronchitis virus (IBV) causes severe economic losses worldwide. IBV has a broad tissue distribution with different viral loads in different tissues. Additionally, IBV can induce apoptosis in infected cells. Aims: The present study aimed to evaluate the role of the genetic background of chickens in viral load and the expression level of apoptotic genes in different tissues of two hybrids of commercial broiler chickens (Ross 308 and Cobb 500) challenged with IBV. Methods: Chickens at 21 days of age were nasally challenged with 200 μL of allantoic fluid containing 104 EID50/ml of Iranian variant-2-like IBV (IS/1494). The expression level of apoptotic genes (Fas, FasL, Bax, and Bcl-2) in the tracheal and renal tissues and the amount of viral load in the tracheal, renal, and cloacal swab samples were investigated two, five, and seven days after IBV infection by RT-qPCR assay. Results: The amount of viral load and apoptotic the expression level of apoptotic genes in the tracheal (two and five days after infection) and renal samples (seven days after infection) were significantly higher in the Ross challenged group than in the Cobb challenged group. Furthermore, no difference was observed in the cloaca viral load on sampling days. Conclusion: To our knowledge, this is the first report that evaluated the role of the chickens’ genetic background in the amount of viral load and the expression level of apoptotic genes against IBV. Further studies are needed to investigate the pathogenic characteristics of IBV in Ross 308 and Cobb 500 chickens.


Main Subjects

Ampomah, PB and Lim, LH (2020). Influenza A virus-induced apoptosis and virus propagation. Apoptosis. 25: 1-11.
Bande, F; Arshad, SS; Omar, AR; Bejo, MH; Abubakar, MS and Abba, Y (2016). Pathogenesis and diagnostic approaches of avian infectious bronchitis. Adv. Virol., 2016: 4621659-4621669.
Barjesteh, N; O’Dowd, K and Vahedi, SM (2020). Antiviral responses against chicken respiratory infections: focus on avian influenza virus and infectious bronchitis virus. Cytokine. 127: 154961-154971.
Belkasmi, SF; Fellahi, S; Touzani, CD; Faraji, FZ; Maaroufi, I; Delverdier, M; Guérin, JL; Fihri, OF; El Houadfi, M and Ducatez, MF (2020). Co-infections of chickens with avian influenza virus H9N2 and Moroccan Italy 02 infectious bronchitis virus: effect on pathogenesis and protection conferred by different vaccination programmes. Avian Pathol., 49: 21-28.
Callison, SA; Hilt, DA; Boynton, TO; Sample, BF; Robison, R; Swayne, DE and Jackwood, MW (2006). Development and evaluation of a real-time Taqman RT-PCR assay for the detection of infectious bronchitis virus from infected chickens. J. Virol. Methods. 138: 60-65.
Cavanagh, D (2007). Coronavirus avian infectious bronchitis virus. Vet. Res., 38: 281-297.
Chen, Z; Zhu, Y; Ren, Y; Tong, Y; Hua, X; Zhu, F; Huang, L; Liu, Y; Luo, Y and Lu, W (2011). Hepatitis C virus protects human B lymphocytes from Fas-mediated apoptosis via E2-CD81 engagement. PLoS One. 6: e18933.
Chhabra, R; Kuchipudi, SV; Chantrey, J and Ganapathy, K (2016). Pathogenicity and tissue tropism of infectious bronchitis virus is associated with elevated apoptosis and innate immune responses. Virology. 488: 232-241.
Clarke, P and Tyler, KL (2009). Apoptosis in animal models of virus-induced disease. Nat. Rev. Microbiol., 7: 144-155.
De Wit, J; Cazaban, C; Dijkman, R; Ramon, G and Gardin, Y (2018). Detection of different genotypes of infectious bronchitis virus and of infectious bursal disease virus in European broilers during an epidemiological study in 2013 and the consequences for the diagnostic approach. Avian Pathol., 47: 140-151.
Dolz, R; Vergara-Alert, J; Pérez, M; Pujols, J and Majó, N (2012). New insights on infectious bronchitis virus pathogenesis: characterization of Italy 02 serotype in chicks and adult hens. Vet. Microbiol., 156: 256-264.
Elmore, S (2007). Apoptosis: a review of programmed cell death. Toxicol. Pathol., 35: 495-516.
Fadhilah, A; Kai, T; Lokman, H; Yasmin, N; Hafandi, A; Hasliza, A; Rinalfi, TT and Hezmee, M (2020). Molecular and pathogenicity of infectious bronchitis virus (Gammacoronavirus) in Japanese quail (Coturnix japonica). Poult. Sci., 99: 2937-2943.
Fu, Q; He, C and Mao, Zr (2013). Epstein-Barr virus interactions with the Bcl-2 protein family and apoptosis in human tumor cellsJ. Zhejiang Univ., Sci., B., 14: 8-24.
Ganapathy, K; Wilkins, M; Forrester, A; Lemiere, S; Cserep, T; McMullin, P and Jones, R (2013). QX-like infectious bronchitis virus isolated from proventriculitis in commercial broilers in England. Vet. Rec., 171: 597.
Han, X; Tian, Y; Guan, R; Gao, W; Yang, X; Zhou, L and Wang, H (2017). Infectious bronchitis virus infection induces apoptosis during replication in chicken macrophage HD11 cells. Viruses. 9: 198-211.
Hatakeyama, S; Sakai-Tagawa, Y; Kiso, M; Goto, H; Kawakami, C; Mitamura, K; Sugaya, N; Suzuki, Y and Kawaoka, Y (2005). Enhanced expression of an α2, 6-linked sialic acid on MDCK cells improves isolation of human influenza viruses and evaluation of their sensitivity to a neuraminidase inhibitor. J. Clin. Microbiol., 43: 4139-4146.
Jordan, B (2017). Vaccination against infectious bronchitis virus: a continuous challenge. Vet. Microbiol., 206: 137-143.
Lan, Y; Zhao, K; Wang, G; Dong, B; Zhao, J; Tang, B; Lu, H; Gao, W; Chang, L and Jin, Z (2013). Porcine hemagglutinating encephalomyelitis virus induces apoptosis in a porcine kidney cell line via caspase-dependent pathways. Virus Res., 176: 292-297.
Liao, H; Xu, J and Huang, J (2010). FasL/Fas pathway is involved in dengue virus induced apoptosis of the vascular endothelial cells. J. Med. Virol., 82: 1392-1399.
Liu, C; Xu, H and Liu, D (2001). Induction of caspase-dependent apoptosis in cultured cells by the avian coronavirus infectious bronchitis virus. J. Virol., 75: 6402-6409.
Liu, H; Yang, X; Zhang, Z; Li, J; Zou, W; Zeng, F and Wang, H (2017). Comparative transcriptome analysis reveals induction of apoptosis in chicken kidney cells associated with the virulence of nephropathogenic infectious bronchitis virus. Microb. Pathog., 113: 451-459.
Livak, KJ and Schmittgen, TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 25: 402-408.
Madu, IG; Chu, VC; Lee, H; Regan, AD; Bauman, BE and Whittaker, GR (2007). Heparan sulfate is a selective attachment factor for the avian coronavirus infectious bronchitis virus Beaudette. Avian Dis., 51: 45-51.
Marandino, A; Vagnozzi, A; Craig, MI; Tomás, G; Techera, C; Panzera, Y; Vera, F and Pérez, R (2019). Genetic and antigenic heterogeneity of infectious bronchitis virus in South America: implications for control programmes. Avian Pathol., 48: 270-277.
Najafi, H; Hashemzadeh, M; Madadgar, O; Karimi, V; Farahani, R; Abdollahi, H; Maghsoudsloo, H and Seifouri, P (2016a). Pathogenicity characteristics of an Iranian variant-2 (IS-1494) like infectious bronchitis virus in experimentally infected SPF chickens. Acta Virol., 60: 393-399.
Najafi, H; Langeroudi, AG; Hashemzadeh, M; Karimi, V; Madadgar, O; Ghafouri, SA; Maghsoudlo, H and
Farahani, RK
(2016b). Molecular characterization of infectious bronchitis viruses isolated from broiler chicken farms in Iran, 2014-2015. Arch. Virol., 161: 53-62.
Najimudeen, SM; Hassan, MSH; Cork, SC and Abdul-Careem, MF (2020). Infectious bronchitis coronavirus infection in chickens: multiple system disease with immune suppression. Pathogens. 9: 779-795.
Reed, LJ and Muench, H (1938). A simple method of estimating fifty per cent endpoints. Am. J. Epidemiol., 27: 493-497.
Ren, G; Liu, F; Huang, M; Li, L; Shang, H; Liang, M; Luo, Q and Chen, R (2020). Pathogenicity of a QX-like avian infectious bronchitis virus isolated in China. Poult. Sci., 99: 111-118.
Tan, YX; Tan, TH; Lee, MJR; Tham, PY; Gunalan, V; Druce, J; Birch, C; Catton, M; Fu, NY and Victor, CY (2007). Induction of apoptosis by the severe acute respiratory syndrome coronavirus 7a protein is dependent on its interaction with the Bcl-XL protein. J. Virol., 81: 6346-6355.
Winter, C; Schwegmann-Weßels, C; Cavanagh, D; Neumann, U and Herrler, G (2006). Sialic acid is a receptor determinant for infection of cells by avian infectious bronchitis virus. J. Gen. Virol., 87: 1209-1216.
Yuan, S; Wu, B; Yu, Z; Fang, J; Liang, N; Zhou, M; Huang, C and Peng, X (2016). The mitochondrial and endoplasmic reticulum pathways involved in the apoptosis of bursa of Fabricius cells in broilers exposed to dietary aflatoxin B1. Oncotarget. 7: 65295-65306.
Zegpi, R; Breedlove, C; Gulley, S and Toro, H (2020). Infectious bronchitis virus immune responses in the harderian gland upon initial vaccination. Avian Dis., 64: 92-95.
Zhao, G; Shi, SQ; Yang, Y and Peng, JP (2006). M and N proteins of SARS coronavirus induce apoptosis in HPF cells. Cell Biol. Toxicol., 22: 313-322.
Zhong, Y; Liao, Y; Fang, S; Tam, JP and Liu, DX (2012). Up-regulation of Mcl-1 and Bak by coronavirus infection of human, avian and animal cells modulates apoptosis and viral replication. PLoS One. 7: e30191.