An Experimental Study of Histopathological Lesions Caused by H9N2 Subtype of Avian Influenza Virus and Ornithobacterium Rhinotracheale Co-Infection

Document Type : Avian (Poultry) Health Management


1 Meshginshahr Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran

2 Department of Avian Diseases Research and Diagnosis, Razi Vaccine and Serum Research Institute, Karaj, Iran

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

4 4Department of Pathology, Razi Vaccine and Serum Research Institute, Karaj, Iran

5 Department of Clinical Sciences, Islamic Azad University, Tabriz Branch, Tabriz, Iran


BACKGROUND: Avian influenza virus H9N2 subtype and Ornithobacterium
(ORT) causes histopathological lesions in birds. Yet, there
is not any study on tissue lesions caused by these pathogens co-infection in
specific pathogen free (SPF) chicks. OBJECTIVES: The aim of this study
was to evaluate the type, severity and extent of histopathological lesions
induced by co-infection of H9N2 subtype avian influenza virus and Ornithobacterium
in SPF chickens. METHODS: Sixty, one-day-old SPF
chicks were divided randomly into three groups of twenty. At 21 days, the
chicks in the first group were infected with H9N2 subtype AI virus and the
second group was inoculated with H9N2 subtype AI virus and ORT simultaneously.
The third group was inoculated with PBS as control. Then, three birds from each
group were randomly selected and euthanized and autopsied at 2  till 16 days post-inoculation (DPI). The
tissue samples were collected from trachea, lungs, liver, spleen, thymus, kidneys,
cecal tonsil and bursa of fabricius. They were cut into 5 to 6 μm thickness
sections using paraffin embedding method and were stained by Hematoxylin and
Eosin (H&E). RESULTS: The severity of histopathological lesions in
the second group was higher than first group. Tissue changes were not observed
in control group. CONCLUSIONS: The results of this study showed that
infected chickens with H9N2 subtype AI virus and ORT simultaneously cause
exacerbated histopathological lesions compared to H9N2 subtype of AI virus


Alexander, D. J. (2007) Summary of avian influenza activity in Europe, Asia, Africa, and Australasia, 2002-2006. Avian Dis. 51(s1): 161-166.##
Azizpour, A., Goudarzi, H., Charkhkar, S., Momayez, R., Hablolvarid, M. H. (2014) Experimental study on tissue tropism and dissemination of H9N2 avian influenza virus and Ornithobacterium rhinotracheale co-infection in SPF chickens. J Anim Plan Sci. 24(6):1655-1662.##
Banani, M., Khaki, P., Goudarzi, H., Vand-Yousefi, J., Pourbakhsh, S.A. (2000) Isolation and identification of Ornithobacterium rhinotracheale from a broiler and a pullet flock. Pajouhesh Va Sazandegi. (In Persian) 46: 106-109.##
Banani, M., Momayez, R., Pourbakhsh, S.A., Goodarzi, H., Bahmani Nejad, M.A. (2002) Simulataneous isolation of Ornithobacterium rhinotracheale and avian influenza virus subtype H9H2 from commercial poultry. Iran J Vet Res. 3(2): 100-115. ##
Banani, M., Pourbakhsh, S.A., Ghodsian, N., Karimi, V., Ashtari, A. (2011) The phylogenetic analysis of some Ornithobacterium rhinotracheale isolates from commercial chicken flocks in Iran. GenBank: JF810491.##
Bano, S., Naeem, K., Malik, S.A. (2003) Evaluation of pathogenic potential of avian influenza virus serotype H9H2 in Chickens. Avian Dis. 47(3 Suppl): 817-822. ##
Bijanzad, P., Momayez, R., Bozorgmehrifard, M.H., Hablolvarid, M.H., Pourbakhsh, S.A. (2013) Experimental study on histopathological changes and tissue tropism of iranian infectious bronchitis serotype 793/B-Like virus in spf chickens. J South Afr Vet Assoc. 84 (1): 1 -7.##
Costa-Hurtado, M., Afonso, C.L., Miller, P.J., Spackman, E., Kapczynski, D.R., Swayne, D.E., Shepherd, E., Smith, D., Zsak, A., Pantin-Jackwood. (2014) Virus interference between H7N2 low pathogenic avian influenza virus and lentogenic Newcastle disease virus in experimental co-infections in chickens and turkeys.Vet Res. 45:1##
Goudarzi, H., Azizpour, A., Banani, M., Nouri, A., Seifi, S. (2015) Survey of tissue tropism and dissemination of ORT-R87-7/1387 strain of Ornithobacterium rhinotracheale in SPF chickens. Pajouhesh Va Sazandegi. (In Persian) 27: 2-8. ## 
Hablolvarid, M. H., Sohraby Haghdost, I., Pourbakhsh, S. A., Gholami, M. R. (2003) Study on histopathologic changes in chicken following intravenous inoculation with avian influenza virus a/chicken/Iran/259/1998(H9N2). Arch Razi Inst. 55(1):  41-54.##
Hablolvarid, M. H., Sohraby Haghdost, I., Pourbakhsh, S. A., Gholami, M. R. (2004) Histopathological study of intranasally inoculated a/chicken/Iran/259/1998(H9N2) Influenza Virus in Chicken. Arch Razi Inst. 58: 51-62.##
Hadipour, M.M., Farjadian, S.H., Azad, F., Kamravan, M., Dehghan, A. (2011) Nephropathogenicity of H9N2 avian influenza virus in commercial broiler chickens following intratracheal inoculation. J Anim Vet Adv. 10(13):1706-1710.##
Haghighat-Jahromi, M., Asasi, K., Nili, H., Dadras, H., Shooshtari, A.H. (2008) Coinfection of avian influenza virus (H9n2 Subtype) with infectious bronchitis live vaccine. Arch Virol. 53(4): 651-655.##
Mosleh, N., Dadras, H., Mohammadi, A. (2009) Evaluation of H9N2 avian influenza virus dissemination in various organs of experimentally infected broiler chickens using RT-PCR. Iranian J Vet Res. 10(1): 12-20.##
Mo, I.P., Brugh, M., Fletcher, O. J., Rowland, G. N., Swayne, D.E. (1997) Comparative pathology of chickens experimentally inoculated with avian influenza viruses of low and high pathogenicity. Avian Dis. 41(1): 125-136.##
Naeem, K., Ullah, A., Manvell, R.J., Alexander, D.J. (1999) Avian influenza a subtype H9H2 in poultry in pakistan. Vet Rec. 145(19): 560-565.##
Nili, H., Asasi, K. (2003) Avian influenza (H9n2) outbreak in Iran. Avian Dise. 47(3 Suppl): 828-831.
Pan, Q., Liu, A., Zhang, F., Ling, Y., Ou, C., Hou, N., He, C. (2012) Co-infection of broilers with Ornithobacterium rhinotracheale and H9N2 avian influenza virus. BMC. Vet Res. 8:104. doi:10.1186/1746-6148-8-104. ##
Pazani, J., Vasfi-Marandi, M., Ashrafihelan, J., Marjanmehr, S. H., Ghods, F. (2008) Pathological studies of a / chicken / Tehran / Zmt - 173/99 (H9N2) influenza virus in commercial broiler chickens of Iran. Inter J Poul Scie. 7: 55-63. ## 
Slemons, R. D., Swayne, D. E. (1990) Replication of a waterfowl-origin influenza virus in the kidney and intestine of chickens. Avian Dis. 34(2): 277-284.##
Swayne, D.E., Suarez, D.L., Sims, L.D. (2013) Influenza. In: Diseases of Poultry.  Swayne, D.E., Glisson, J.R., McDougald, L.R., Nolan, L.K., Suarez, D.L., Venugopal, N. (eds.). (13th ed.) Wiley-Blackwell, Ames, USA. p. 181-218.##
Swayne, D. E., Slemons, R. D. (1990) Renal pathology in specific-pathogen-free chickens inoculated with a waterfowl-origin type a influenza virus. Avian Dis. 34(2): 285 -294.##
Swayne, D. E., Slemons, R. D. (1994) Comparative pathology of a chicken-origin and two duck-origin influenza virus isolates in chickens: the effect of route of inoculation. Vet Pathol. 31(2): 237-245.##
Thachil, A.J., Velayudhan, B.T., Shaw, D.P., Halvorson, D.A., Nagaraja, K.V. (2009) Pathogenesis of Ornithobacterium rhinotracheale in egg-laying hens with coexisting infectious bronchitis virus and Escherichia coli infections. J Appl Poult Res. 18:780-788.##
Van Empel, P., Hafez, H.M. (1999) Ornithobacteriumrhinotracheale: a review. Avian Dis. 28: 217- 227.
Vasfi Marandi, M., Bozorgmehri Fard, M. H. (2002) Isolation of H9N2 subtybe of avian influenza viruses during outbreak in chickens in Iran. Iran Biomed J. 6: 13-17.##