BACKGROUND: Avibacterium paragallinarum is a causative agent of infectious coryza (IC), a disease that affects the upper respiratory tracts and paranasal sinuses of chickens, resulting significant economic losses in the poultry industry. The objective of this study was to isolate and identify Av. paragallinarum using bacteriological and molecular methods between February 2022 and April 2024. A total of 74 swab samples were collected from chickens showing ocular and nasal discharges and swelling of the infraorbital sinuses. METHOD: Clinical samples were collected from chickens showing symptoms of IC from six locations of Ethiopia for the isolation and identification of the causative agent. Swab samples from the nasal cavity and cheesy material from the infraorbital sinus were screened using conventional PCR and inoculated onto chocolate agar enriched with 5% sheep blood. Colonies suspected of being Av. paragallinarum were transferred to brain heart agar supplemented with horse serum. Gram staining was used to examine the morphology of bacteria in pure colonies grown on chocolate and brain heart infusion agar. RESULTS: The isolation of Av. paragallinarum on chocolate and brain heart infusion agar resulted in the observation of small, translucent, dewdrop-shaped colonies after 24 h of incubation at 37 °C in a 5% CO CONCLUSION: The current investigation confirmed that the outbreaks were caused by Avibacterium paragallinarum and provided scientific evidence on the presence of different strains of Av. paragallinarum in Ethiopia. This is the first study in Ethiopia to detect and identify Av. paragallinarum from diseased chickens using molecular approaches. Further molecular characterization of locally circulating Av. paragallinarum isolates is recommended to be used as a vaccine strain for the prevention and control of infectious coryza.