This cross-sectional study assessed biosecurity, poultry farm structure, and antibiotic use practices in Côte d’Ivoire between February and July 2024, based on a questionnaire survey administered to poultry farmers and descriptive and analytical statistical analyses. The results show a strong concentration of farms in urban and peri-urban areas, particularly in Abidjan, reflecting an intensification of poultry production oriented toward urban markets. Production systems are dominated by broiler farming (over 70%), followed by layer farming (approximately 28%). Most farms are medium-sized and managed by the owners themselves, with generally intermediate to low levels of education and technical knowledge. The biosecurity assessment reveals an overall inadequate level. Indeed, 58.8% of farms have low biosecurity and 26.1% have no structured biosecurity measures. Preventive measures remain poorly implemented, particularly footbaths (8.4%), fencing (30.2%), and farm isolation (42.9%), thereby increasing the risk of pathogen transmission and spread. Antibiotic use is high and often poorly regulated. About 62.5% of farmers report systematic antibiotic use, while 99.5% do not perform any laboratory diagnosis before treatment. Practices are largely empirical, with treatment durations generally ranging from 3 to 5 days. The main antibiotic classes used are tetracyclines (33%) and aminoglycosides (30%), followed by trimethoprim and fluoroquinolones. These findings highlight the need to strengthen biosecurity, technical support, and the rational use of antimicrobials within a One Health approach.