| Summary: | Disease prevalence and seasonal outbreaks are challenging the poultry industry in Ethiopia. Proper and sustainable implementation of biosecurity practices is important to reverse such problems. This study was conducted in commercial poultry farms in two zones of Ethiopia to investigate farm characteristics, implementation of biosecurity practices, and biosecurity status (BS) using a structured questionnaire. The variables were grouped into three biosecurity factors, including conceptual, structural, and operational biosecurity, based on their homogeneity. Descriptive and inferential statistics were used to summarize the results. Most commercial farms were owned by males (69.7%). The majority of the farms (40.3%) were located at a distance <50 m from residential areas. Farm owners do not provide biosecurity training to their employees (68.8%), which results in poor biosecurity implementation. The mean conceptual, structural, and operational BS were 50.4 ± 11.62, 63.27 ± 10.51, and 44.69 ± 13.04, respectively, indicating operational biosecurity measurements were less implemented. Overall, the BS indicated that 40.7% of the farms have BS < 50% questing for interventions. Farm characteristics and biosecurity measurements were positively associated with BS, which shows substantial room for improvement. Owners’ education, occupation, experience, farm flock size, and training were significantly associated with BS (<i>p</i> < 0.05). A disease prevention strategy through biosecurity improvement is an economical means for controlling poultry disease prevalence.
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