Prevalence and risk factors of Influenza Avian Virus in backyard pigeons, ducks, and chickens in Toba Tek Singh District, Pakistan

Influenza Avian virus (IAV) is a zoonotic pathogen that can be transmitted from birds to humans. Multiple IAV pandemics have had a devastating impact on the poultry industry and backyard birds (including ducks, chickens, and pigeons) worldwide, notably in Europe, United States, Africa, and Asia. In Pakistan, numerous outbreaks of H7, H5, and H9 subtypes have been documented in both commercial and rural areas, resulting in significant financial losses. However, the epidemiological status of various IAV subtypes in backyard birds in rural areas remains largely unknown. This study aimed to evaluate the prevalence of IAV and associated risk factors among domesticated birds in the Toba Tek Singh District, Pakistan. A cross-sectional study was conducted between 2017 and 2019 using multistage cluster sampling approach. Pooled tracheal and cloacal swab samples were collected and tested for IAV. Positive pooled swab samples were subsequently evaluated at the individual level. RNA was extracted using theTrizol method, followed by multiplex RT‒PCR with specific primers and probes to detect the IAV M-gene and its subtypes. Statistical analysis was performed using a multivariable logistic regression model. Overall, the prevalence of IAV in backyard chickens, pigeons, and ducks was 13.4%, 7.7%, and 11.4%, respectively. The most commonly detected IAV subtypes included H7, H9, and HA/Untyped. No statistically significant difference (p > 0.05) in IAV prevalence was observed across cities for any bird species. In the multivariable analysis, species type (particularly chickens and pigeons) was significantly associated with IAV prevalence, while fighting cocks showed a borderline association. Enhanced surveillance, improved biosecurity protocols, targeted educational initiatives, and the adoption of better farming practices are recommended to mitigate IAV transmission and safeguard both poultry production and public health in Pakistan.