Avian influenza (AI) is a global health obstacle of critical concern as novel viruses are capable of initiating a pandemic. Recent spillover events of AI into human populations have occurred at human-poultry food system interfaces. As Nepal´s poultry sector transitions to more intensified commercial production systems, it is important to examine the epidemiology of AI and the knowledge, attitudes and practices (KAP) of poultry sector workers. We conducted a cross-sectional KAP study utilizing a structured survey to interview 150 commercial poultry farmers in Chitwan District, Nepal. All commercial poultry farmers had knowledge of AI previous to the study and the majority farmers were able to identify farm-farm and poultry-human transmission mechanisms of AI. Farmers had more knowledge surrounding poultry AI symptoms as compared to human AI symptoms. Most farmers believe that AI is serious, contagious and a threat to everyone, yet only half believe it can be prevented. Individual-level personal protective equipment (PPE) uptake, such as facemask, glove and boot usage, on the enrolled farms was low and farm-level biosecurity practices varied greatly. Nine commercial poultry farms (6%) self-reported having an HPAI outbreak and 60 farms (40%) self-reported having an LPAI outbreak in the past 5 years. Layer farms had higher odds (OR: 5.4, 95% CI: 2.3-12.8) of self-reported LPAI as compared broiler farms. Poultry sector farmers face multiple obstacles when attempting to report AI to government authorities such as the fear of flock culling and the perceived lack of monetary compensation for culling. Our study provides updated KAP surrounding AI of farmers and self-reported AI farm-level epidemiology in Nepal´s highest density commercial poultry production district. Commercial poultry farmers are fairly knowledgeable on AI, but do not take further protective practice efforts to implement their knowledge and prevent AI. Due to the potential role that human-poultry interfaces may play in AI emergence, it is critical to collaborate with the commercial poultry industry when planning and conducting AI pandemic preparedness mechanisms.