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India's livestock sector has been facing significant losses due to episodes of disease outbreaks since time immemorial. Hence, biosecurity measures are very important to maintain and improve animal health along with prevention of disease outbreak. Keeping these facts into consideration, the study was proposed with an objective to assess the existing biosecurity practices adopted by the commercial dairy, pig and poultry farms. The current study was undertaken in the state of Uttar Pradesh as it is the leading state in milk and meat production. A total of 120 farmers were selected randomly including 40 each practicing commercial dairy, pig and poultry farming. An ex-post facto research methodology was used with face-to-face interview and observation to collect data. The biosecurity practices were assessed under seven dimensions such as, location and design of farm, restricted access, isolation and quarantine, cleaning and disinfection, management of feed and water, disposal of carcass, manure and waste, and health management. Results elicited that about 50% of the farmers had medium level of adoption who adopted 18-34 practices out of 51 practices. The average overall adoption score was 34.17 out of 51 (67%) which makes an overall adoption gap of 33%. Maximum adoption gap was seen in case of restricted access (43%) whereas minimum gap in adoption was seen in case of management of feed and water (27%). Pig and poultry farmers showed significantly higher biosecurity measures than dairy farmers (p < 0.05). The more significant contributors to the adoption of biosecurity measures were the level of knowledge of the farmers (p < 0.01). Other factors such as education, income, herd/flock size, Information and Communication Technology utilization, number of trainings also had a significant contribution (p < 0.05) in actual implementation of biosecurity. Hence, better understanding of these measures among the farmers must be ensured by hands on training along with proper demonstration of various procedures involved in maintaining farm biosecurity is need of the hour., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)