Document Type : Original Article


1 Department of Poultry and obstetrics Sciences and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Poultry and Sciences and Research Branch, Islamic Azad University, Tehran, Iran

3 Department of Poultry and Sciences and Research Branch, Islamic Azad University, Tehran, Iran.

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

5 Department of Basic Sciences, Faculty of Veterinary Medicine, Kazerun Branch, Islamic Azad University, Kazerun, Iran


The aim of this study was to provide information on the molecular characteristic and the phylogenic relationship of infectious bronchitis viruses (IBV) strains in Bushehr province in comparison to other strains reported in the Middle East. Samples were collected from broiler flocks in Bushehr province during 2014 - 2015. These flocks had respiratory problems such as gasping, sneezing and bronchial rales. A number of 135 tracheal swabs were taken from fifteen flocks (nine swabs per flock). Each three swabs collected from each flock were pooled in one tube (finally, we had three tubes for each flock). The samples were subjected to reverse transcription polymerase chain reaction (RT-PCR). The PCR products of positive samples were analyzed by sequencing of a (392 bp) segment of the spike gene and the related results were compared with the other IBV sequences in GenBank database. Samples from twelve farms (80.0%) were found to be positive. The viruses from seven farms (46.6%) were identified as field viruses closely related to variant 2. The viruses from three farms (20.0%) were characterized as Mass type and were related to vaccine strains. Two different IB viruses (variant 2 and Mass) were detected in samples from two farms (13.3%). The variant 2 genotype detected in Bushehr had high similarity to variant 2 reported from the Middle East.These variants displayed homologies ranging from 72.9% to 76.5%, and 78.8% to 80.0% with H120 and 4/91, respectively. It is necessary to design vaccination program of poultry farms using IBV strains circulating in the region.


Main Subjects


    1. Butcher GD, Shapiro DP, Miles RD. Infectious bronchitis virus: Classical and variant strains. One of a series of the Veterinary medicine-large animal clinical sciences department, Florida cooperative extension service. UF/IFAS Extension 2011; VM127.
    2. Ignjatovic J, Sapats S. Avian infectious bronchitis virus. Rev Sci Tech 2000; 19: 493-508.
    3. Pohuang T, Chansiripornchai N, Tawatsin A, et al. Detection and molecular characterization of infectious bronchitis virus isolated from recent outbreaks in broiler flocks in Thailand. J Vet Sci 2009; 10: 219-223.
    4. Cavanagh D. Severe acute respiratory syndrome vaccine development experiences of vaccination against avian infectious bronchitis coronavirus. Avian Pathol 2003; 3(2): 567-582.
    5. Spaan W, Cavanagh D, Horzinek MC. Coronaviruses: Structure and genome expression. J Gen Virol 1988; 69: 2939-2952.
    6. Sutou S, Sato S, Okabe T, et al. Cloning and sequencing of genes encoding structural proteins of avian infectious bronchitis virus. Virology 1988; 165: 589-595.
    7. Cavanagh D. Coronavirus IBV: Structural characterization of the spike protein. J Gen Virol 1983; 64: 2577-2583.
    8. Cavanagh D. The coronavirus surface glycoprotein. In: Siddel SG (Ed). The Coronaviridae. New York, USA: Plenum Press 1995; 73- 103.
    9. Cavanagh D, Davis PJ, Mockett AP. Amino acids within hypervariable region 1 of avian coronavirus IBV (Massachusetts serotype) spike glycoprotein are associated with neutralization epitopes. Virus Res 1988; 11: 141-150.
    10. Koch G, Hartog L, Kant A, et al. Antigenic domains on the peplomer protein of avian infectious bronchitis virus: correlation with biological functions. J Gen Virol 1990; 71. 1929-1935.
    11. Moore KM, Jackwood MW, Hilt DA. Identification of amino acids involved in a serotype and neutralization specific epitope within the S1 subunit of avian infectious bronchitis virus. Arch Virol 1997; 142: 2249-2256.
    12. Cavanagh D. Structural polypeptides of coronavirus IBV. J Gen Virol 1981; 53: 93.
    13. Stern DF, Sefton BM. Coronavirus proteins: Structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins. J Virol 1982; 44: 804-812.
    14. Cavanagh D. Structural Characterization of IBV Glycoproteins. In: Rottier P.J.M., van der Zeijst B.A.M., Spaan W.J.M., Horzinek M.C. (Eds) Molecular Biology and Pathogenesis of Coronaviruses. Advances in Experimental Medicine and Biology, vol 173. Springer, Boston, MA, 1984; 95-108.
    15. Mockett APA, Cavanagh D, Brown TDK. Monoclonal antibodies to the S1 spike and membrane proteins of avian infectious bronchitis virus coronavirus strain Massachusetts M41. J Gen Virol 1984; 65: 2281-2286.
    16. Cavanagh D, Davis PJ. Coronavirus IBV: Removal of spike glycopolypeptide S1 by urea abolishes infectivity and hemagglutination but not attachment to cells. J Gen Virol 1986; 67: 1443-1448.
    17. Holmes KV, Lai MM. Coronaviridae: The virus and their replication. In: Fields BN, Knipe DM, Howley PM (Eds). Fundamental virology. 4th ed. Philadelphia, USA: Lippincott-Raven 2001; 1163-1203.
    18. Cavanagh D, Davis PJ, Cook JKA, et al. Location of the amino-acid differences in the S1 spike glycoprotein subunit of closely related serotypes of infectious bronchitis virus. Avian Pathol 1992; 21: 33-43.
    19. Kant A, Koch G, Van Roozelaar DG, et al. Location of antigenic sites defined by neutralizing monoclonal antibodies on the S1 avian infectious bronchitis virus glycopolypeptide. J Gen Virol 1992; 73: 591-596.
    20. Bochkov YA, Batchenko GV, Shcherbakova LO, et al. Molecular epizootiology of avian infectious bronchitis in Russia. Avian Pathol 2006; 35: 379-393.
    21. Ammayappan A, Upadhyay C, Gelb J, et al. Complete genomic sequence analysis of infectious bronchitis virus Ark DPI, strain and its evolution by recombination. Virol J 2008; 5: 157.
    22. Cavanagh D. Coronavirus avian infectious bronchitis virus. Vet Res 2007; 38 (2): 281-297.
    23. Cook JK, Orbell SJ, Woods MA, et al. A survey of the presence of a new infectious bronchitis virus designated 4/91 (793B). Vet Rec 1996; 13 (8): 178-180.
    24. Liu SW, Zhang QX, Chen JD, et al. Genetic diversity of avian infectious bronchitis coronavirus strains isolated in China between 1995 and 2004. Arch Virol 2006; 151 (6): 1133-1148.
    25. Aghakhan SM, Abshar N, Fereidouni SRN, et al. Studies on avian viral infections in Iran. Arch Razi Ins 1997; 44: 1-10.
    26. Vasfi Marandi M, Bozogmehri Fard MH. Isolation and identification of infectious bronchitis viruses in chickens between 1997-2000 in Iran. J Vet Res 2001; 56: 119-124.
    27. Hosseini H, Fard MH, Charkhkar S, et al. Epidemiology of avian infectious bronchitis virus genotypes in Iran (2010-2014). Avian Dis 2015; 59(3): 431-435.
    28. Najafi H, Langeroudi AG, Hashemzadeh M, et al. Molecular characterization of infectious bronchitis viruses isolated from broiler chicken farms in Iran. 2014-2015. Arch Virol 2016; 161(1):53-62.
    29. Homayounimehr A, Pakbin A, Momayyez R, et al. Detection and identification of infectious bronchitis virusby RT-PCR in Iran. Trop Anim Health Prod 2016; 48(5): 973-978.
    30. Jones RC, Worthington KJ, Gough, RE. Detection of the Italy 02 strain of infectious bronchitis virus in the UK. Vet Rec 2005; 156-260.
    31. Tamura K, Peterson D, Peterson N, et al. MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Mol Biol Evol. 2011; 28(10):2731-2739.
    32. Adznar AK, Cough RE, Haydon D, et al. Molecular analysis of the 793/B serotype of IBV in Great Britain. Avian Pathol 1997; 26: 625-640.
    33. Cavanagh D, Ellis MM, Cook JKA. Relationship between sequence variation in the S1 spike protein of infectious bronchitis virus and the extent of cross-protection in vivo. Avian Pathol 1997; (26): 63-74.
    34. Katarzyna D, Krzysztof S, Zenon M. Molecular studies on infectious bronchitis virus isolated in Poland. Bull Vet Inst Pulawy 2007; 51: 449-452.
    35. de Wit JJ, Cook JK. Factors influencing the outcome of infectious bronchitis vaccination and challenge experiments. Avian Pathol 2014; 43: 485-497.
    36. Ganapathy K, Ball C, Forrester A. Genotypes of infectious bronchitis viruses circulating in the Middle East between 2009 and 2014. Virus Res 2015; 210: 198-204.
    37. Mahmood ZH, Sleman R, Uthman AU. Isolation and molecular characterization of Sul/01/09 avian infectious bronchitis virus indicates the emergence of a new genotype in the Middle East. Vet Microbiol 2011; 150: 21-27.
    38. Seger W, Ghalyanchi Langeroudi A, Karimi V, et al. Genotyping of infectious bronchitis viruses from broiler farmsin Iraq during 2014-2015. Arch Virol 2016; 161(5):1229-1237.
    39. Abdel-Moneim AS, Afifi MA, El-Kady MF. Emergence of a novel genotype of avian infectious bronchitis virus in Egypt. Arch Virol 2012; 157: 2453-2457.
    40. Ababneh M, Dalab AE, Al saad S. Presence of infectious bronchitis virus strain CK/CH/LDL/97I in the Middle East. ISRN Vet Sci 2012; 1-6.
    41. Kahya S, Coven F, Temelli S, et al. Presence of IS/1494/06 genotype-related infectious bronchitis virus in breeder and broiler flocks in Turkey. Ankara Univ Vet Fak Derg 2013; 60: 27-31.
    42. Selim K, Arafa AS, Hussein HA. Detection and molecular characterization of infectious bronchitis viruses isolated from broiler and layer chicken farms in Egypt during 2012. Int J Virol 2014; 10(1): 70-76.
    43. Awad F, Baylis M, Ganapathy K. Detection of variant infectious bronchitis viruses in broiler flocks in Libya. Int J Vet Sci Med 2014; (2): 78-82.
    44. Meir R, Rosenblut E, Perl S, et al. Identification of a novel nephropathogenic infectious bronchitis virus in Israel. Avian Dis 2004; 48: 635-641.
    45. Hashemzadeh M, Karimi V, Masoudi S, et al. Phylogenetic study of Iranian infectious bronchitis virus isolates during 2010 - 2011 using glycoprotein S1 gene. J Vet Res 2013; (68):135-141.
    46. Seyfi Abad Shapouri MR, Mayahi M, Assasi K, et al. A survey of the prevalence of infectious bronchitis virus type 4/91 in Iran. Acta Vet Hung 2004; 52: 163-166.
    47. Rauf HS. Investigation and molecular characterization of avian infectious bronchitis virus in suspected broiler farms in Slemani Governorate. MS Thesis, University of Sulaimani. Sulaymaniyah, Iraq: 2014.