Molecular characterization of common respiratory viral infections in broilers in Al-Hassa, Eastern Province, Saudi Arabia

Newcastle disease virus (NDV), Infectious bronchitis virus (IBV) and H9N2 avian influenza virus (AIV) are the most significant respiratory pathogens. Several outbreaks have been reported frequently in Saudi Arabia despite of the intensive use of vaccination programs. Ten broiler flocks suffering from complicated respiratory disease were subjected to necropsy and samples were collected for molecular identification of involved viruses and pathogenic bacterial infections. Seven out of the ten flocks were positive for single or combined viral infection. The seven flocks were positive for HA test. All positive samples were subjected for molecular identification, three flocks were positive for single NDV while one flock was positive for both NDV and IBV. Three flocks were positive for H9N2 and IBV. Three flocks were negative for viral infection and subjected for pathogenic bacteria detection. The sequence analysis of F gene of NDV revealed that all isolates exhibited the cleavage site ( 112 RRQKRF 117 ) of virulent NDVs. The neuraminidase-haemagglutinin gene revealed that all AI isolates belonged to H9N2 low pathogenic avian influenza virus (LPAIV) subtype. The S1 gene of IBV showed that one isolate was closely related to the Connecticut IBV strain while another isolate belonged to the H120 IBV strain. In conclusion, combined pathogens causing respiratory infection in broilers is very common, which leads to the increase in disease severity and economic losses. The continuous detection of virulent NDV as well as IBV from broiler flocks emphasize the need to review the vaccination strategies based on further epidemiological and molecular studies.

[1]  A. El-Sanousi,et al.  Molecular and antigenic traits on hemagglutinin gene of avian influenza H9N2 viruses: Evidence of a new escape mutant in Egypt adapted in quails. , 2017, Research in veterinary science.

[2]  Jianqiang Ye,et al.  Detection of influenza A virus from live-bird market poultry swab samples in China by a pan-IAV, one-step reverse-transcription FRET-PCR , 2016, Scientific Reports.

[3]  I. Kiss,et al.  Successive occurrence of recombinant infectious bronchitis virus strains in restricted area of Middle East , 2016, Virus evolution.

[4]  D. Damena,et al.  Characterization of Newcastle disease virus isolates obtained from outbreak cases in commercial chickens and wild pigeons in Ethiopia , 2016, SpringerPlus.

[5]  A. Ghalyanchilangeroudi,et al.  Genotyping of infectious bronchitis viruses from broiler farms in Iraq during 2014-2015 , 2016, Archives of Virology.

[6]  A. Omar,et al.  Pathogenesis and Diagnostic Approaches of Avian Infectious Bronchitis , 2016, Advances in virology.

[7]  R. Jafari,et al.  Molecular characterization and phylogenetic study of the fusion genes of Newcastle disease virus from the recent outbreaks in Ahvaz, Iran , 2016, VirusDisease.

[8]  T. Peacock,et al.  Antigenic mapping of an H9N2 avian influenza virus reveals two discrete antigenic sites and a novel mechanism of immune escape , 2016, Scientific Reports.

[9]  K. Ganapathy,et al.  Genotypes of infectious bronchitis viruses circulating in the Middle East between 2009 and 2014. , 2015, Virus research.

[10]  Ali Ahmed Abdelkader,et al.  Phylogentic analysis of recent infectious bronchitis virus isolates from broiler chicken farms in Kafrelsheikh, Egypt , 2015 .

[11]  F. Chen,et al.  Isolation and phylogenetic analysis of hemagglutinin gene of H9N2 influenza viruses from chickens in South China from 2012 to 2013 , 2015, Journal of veterinary science.

[12]  Hung-Jen Liu,et al.  Characterization of emerging Newcastle disease virus isolates in China , 2015, Virology Journal.

[13]  S. A. Pourbakhsh,et al.  Molecular characterization of H9N2 avian influenza viruses isolated from vaccinated broiler chickens in northeast Iran , 2015, Tropical Animal Health and Production.

[14]  M. Thrusfield,et al.  Reassortant Avian Influenza A(H9N2) Viruses in Chickens in Retail Poultry Shops, Pakistan, 2009–2010 , 2015, Emerging infectious diseases.

[15]  A. Arafa,et al.  Isolation and characterization of avian influenza from different species of ducks in delta region , 2015 .

[16]  A. Wajid,et al.  Identification of new sub-genotypes of virulent Newcastle disease virus with potential panzootic features. , 2015, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[17]  A. Rahimian,et al.  Phylogenetic characterization of the fusion genes of the Newcastle disease viruses isolated in Fars province poultry farms during 2009-2011 , 2014, Veterinary research forum : an international quarterly journal.

[18]  M. Baylis,et al.  Detection of variant infectious bronchitis viruses in broiler flocks in Libya , 2014 .

[19]  E. H. Chowdhury,et al.  Full-genome analysis of avian influenza virus H9N2 from Bangladesh reveals internal gene reassortments with two distinct highly pathogenic avian influenza viruses , 2014, Archives of Virology.

[20]  R. Webster,et al.  Genesis of avian influenza H9N2 in Bangladesh , 2014, Emerging Microbes & Infections.

[21]  M. Najafi,et al.  Molecular characterization and phylogenetic study of velogenic Newcastle disease virus isolates in Iran , 2013, Virus Genes.

[22]  P. Collins,et al.  Newcastle Disease Virus Fusion Protein Is the Major Contributor to Protective Immunity of Genotype-Matched Vaccine , 2013, PloS one.

[23]  Guo Zhao,et al.  Full Genome Sequence of a Natural Reassortant H9N2 Avian Influenza Virus Isolated from Domestic Ducks in Jiangsu Province, China , 2013, Genome Announcements.

[24]  M. Radwan,et al.  Isolation and molecular characterization of Newcastle disease virus genotypes II and VIId in Egypt between 2011 and 2012 , 2013, Virus Genes.

[25]  A. Anjum,et al.  Genetic diversity of Newcastle disease virus in Pakistan: a countrywide perspective , 2013, Virology Journal.

[26]  I. Brown,et al.  Phylogenetic and molecular characteristics of Eurasian H9 avian influenza viruses and their detection by two different H9-specific RealTime reverse transcriptase polymerase chain reaction tests. , 2013, Veterinary microbiology.

[27]  M. Jackwood,et al.  Evaluation of Infectious Bronchitis Virus Arkansas-Type Vaccine Failure in Commercial Broilers , 2013, Avian diseases.

[28]  M. Hedayati,et al.  Detection of infectious bronchitis virus serotypes by reverse transcription polymerase chain reaction in broiler chickens , 2013, SpringerPlus.

[29]  M. Ayim-Akonor,et al.  Molecular Based Survey of Pathogens Associated with Respiratory Disease Outbreaks in Broiler Chickens in Accra , 2013 .

[30]  A. Panshin,et al.  Genetic characterization of HA gene of low pathogenic H9N2 influenza viruses isolated in Israel during 2006–2012 periods , 2013, Virus Genes.

[31]  A. Ghram,et al.  S1 gene sequence analysis of new variant isolates of avian infectious bronchitis virus in Tunisia. , 2012, Veterinary medicine.

[32]  A. Bozorgi,et al.  Molecular characterization and phylogenetic analysis of hemagglutinin and neuraminidase genes of H9N2 avian influenza viruses isolated in Iran in 1999 and 2009 , 2012 .

[33]  Jinliang Wang,et al.  A multiplex RT-PCR assay for detection and differentiation of avian H3, H5, and H9 subtype influenza viruses and Newcastle disease viruses. , 2012, Journal of virological methods.

[34]  Youn-Jeong Lee,et al.  Genetic evolution of the H9N2 avian influenza virus in Korean poultry farms , 2012, Virus Genes.

[35]  A. Barin,et al.  Full-length characterization and phylogenetic analysis of hemagglutinin gene of H9N2 virus isolated from broilers in Iran during 1998–2007 , 2012, Comparative Clinical Pathology.

[36]  R. H. Al-Rifai,et al.  Ornithobacterium rhinotracheale and Mycoplasma synoviae in broiler chickens in Jordan. , 2011, Revue scientifique et technique.

[37]  E. Holmes,et al.  Phylogeography and Evolutionary History of Reassortant H9N2 Viruses with Potential Human Health Implications , 2011, Journal of Virology.

[38]  S. Samal,et al.  Sequence analysis of fusion protein gene of Newcastle disease virus isolated from outbreaks in Egypt during 2006 , 2011, Virology Journal.

[39]  Ping Liu,et al.  Genetic analysis revealed LX4 genotype strains of avian infectious bronchitis virus became predominant in recent years in Sichuan area, China , 2010, Virus Genes.

[40]  H. Amer,et al.  First year of the Highly Pathogenic Avian Influenza H5N1 outbreak in Egypt: rapid antigenic/molecular diagnosis and virus isolation. , 2010 .

[41]  K. Asasi,et al.  Natural co-infection caused by avian in fl uenza H 9 subtype and infectious bronchitis viruses in broiler chicken farms , 2010 .

[42]  S. Malik,et al.  Multiplex polymerase chain reaction for the detection and differentiation of avian influenza viruses and other poultry respiratory pathogens. , 2009, Poultry science.

[43]  D. A. Roussan,et al.  Avian influenza virus H9 subtype in poultry flocks in Jordan. , 2009, Preventive veterinary medicine.

[44]  S. Gharaibeh Pathogenicity of an Avian Influenza Virus Serotype H9N2 in Chickens , 2008, Avian diseases.

[45]  C. Ros,et al.  Molecular epizootiology of infectious bronchitis virus in Sweden indicating the involvement of a vaccine strain , 2002, Avian pathology : journal of the W.V.P.A.

[46]  P. Jørgensen,et al.  Detection and strain differentiation of infectious bronchitis virus in tracheal tissues from experimentally infected chickens by reverse transcription-polymerase chain reaction. Comparison with an immunohistochemical technique. , 1999, Avian pathology : journal of the W.V.P.A.

[47]  J. Gelb,et al.  Differentiation of infectious bronchitis virus serotypes using polymerase chain reaction and restriction fragment length polymorphism analysis. , 1993, Avian diseases.