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SEROLOGICAL AND MOLECULAR DETECTION OF HIGHLY PATHOGENIC AVIAN INFLUENZA OF LAYER HENS CIRCULATING IN DIYALA GOVERNORATE

Authors :

Zainab Abd Awan Al-Talabani¹

1- Department of Microbiology, Collage of Veterinary Medicine, University of Diyal.

Keywords :

avian influenza virus (AIV) H5N8،ELISA،layer chickens،GenBank،RT-PCR

Abstract :

This study was conducted at the College of Veterinary Medicine, University of Diyala, to evaluate the seroprevalence and molecular detection of avian influenza virus (AIV) H5N8 in layer chickens. Samples were collected from eight commercial flocks displaying severe respiratory symptoms, facial edema, followed by cyanosis in all parts of unfeather skin predominantly the wattles comb. Serological analysis was performed using ELISA on 364 serum samples to detect IgG antibodies specific to AIV H5N8. The results showed a significant increase of IgG antibodies in such serum samples at days 70 and 200 of age. Additionally, 48 postmortem tissue samples from trachea, lung, and liver were collected from birds at 200 days of age for RT-PCR analysis by using specific primers for H5N8 strain. Results indicated that 32 out of 48 samples (66.6%) were positive for H5N8, confirmed by sequencing of the 320 bp amplicon. The local H5N8 strain exhibited 99% sequence identity with reference sequences and was registered in GenBank under accession number (ON247929.1.). Phylogenetic analysis showed that the local sample (ON247929.1) was closely related to reference isolates from the NCBI, including MW961428.1, MW961444.1, MW961476.1, MW961436.1, and MW961484.1, which were deposited from Nigeria.