This study examines farmers' knowledge and practices regarding antibiotic use in livestock production in Kenya, with a focus on antimicrobial resistance (AMR). It assesses farmers' understanding of AMR, safe antibiotic use, and factors influencing their knowledge and practices. A cross-sectional quantitative survey was conducted with 319 farming households in five counties in Kenya. Multivariate regression analysis was used to identify factors influencing farmers' knowledge and practices. About 80% of households use antibiotics in their livestock, with 58% administering them themselves. Most farmers buy antibiotics without a prescription, and antibiotics are used for both therapeutic and non-therapeutic purposes, particularly as growth promoters in poultry. Farmers report shorter withdrawal periods than recommended, and while most farmers are aware of AMR, there are still knowledge gaps. The number of livestock owned was the most significant factor positively influencing farmers' knowledge of AMR and safe use. Kenya has made progress in creating awareness about the risks and requirements of antibiotic use in livestock. However, farmers' antibiotic practices continue to pose a significant risk of further AMR development. Knowledge alone is not sufficient to ensure behavioral change. The study recommends effective policy interventions, research on alternatives like probiotics and vaccines, public awareness campaigns, and strengthened collaboration among stakeholders to address the complex issue of AMR. The study highlights the need for targeted education and awareness-raising to address specific knowledge gaps and misconceptions about antibiotic use and AMR. It also emphasizes the importance of regulatory mechanisms to ensure proper antibiotic control and monitoring. Promoting research into alternatives to antibiotics is essential to reduce reliance on them in animal production. Addressing the complex factors influencing antibiotic use in Kenyan livestock farming requires a multifaceted approach involving education, regulation, and targeted interventions to ensure the sustainable and responsible use of antibiotics, safeguarding animal, human, and environmental health.This study examines farmers' knowledge and practices regarding antibiotic use in livestock production in Kenya, with a focus on antimicrobial resistance (AMR). It assesses farmers' understanding of AMR, safe antibiotic use, and factors influencing their knowledge and practices. A cross-sectional quantitative survey was conducted with 319 farming households in five counties in Kenya. Multivariate regression analysis was used to identify factors influencing farmers' knowledge and practices. About 80% of households use antibiotics in their livestock, with 58% administering them themselves. Most farmers buy antibiotics without a prescription, and antibiotics are used for both therapeutic and non-therapeutic purposes, particularly as growth promoters in poultry. Farmers report shorter withdrawal periods than recommended, and while most farmers are aware of AMR, there are still knowledge gaps. The number of livestock owned was the most significant factor positively influencing farmers' knowledge of AMR and safe use. Kenya has made progress in creating awareness about the risks and requirements of antibiotic use in livestock. However, farmers' antibiotic practices continue to pose a significant risk of further AMR development. Knowledge alone is not sufficient to ensure behavioral change. The study recommends effective policy interventions, research on alternatives like probiotics and vaccines, public awareness campaigns, and strengthened collaboration among stakeholders to address the complex issue of AMR. The study highlights the need for targeted education and awareness-raising to address specific knowledge gaps and misconceptions about antibiotic use and AMR. It also emphasizes the importance of regulatory mechanisms to ensure proper antibiotic control and monitoring. Promoting research into alternatives to antibiotics is essential to reduce reliance on them in animal production. Addressing the complex factors influencing antibiotic use in Kenyan livestock farming requires a multifaceted approach involving education, regulation, and targeted interventions to ensure the sustainable and responsible use of antibiotics, safeguarding animal, human, and environmental health.