<p>Excessive antibiotic use in cattle farming contributes to residues in animal products and the emergence of antimicrobial resistance (AMR). This study assessed the knowledge, attitudes, and practices (KAP) of large animal farmers regarding antibiotic use in Bangladesh’s major dairy and beef fattening regions. Data were collected through a pre-tested questionnaire and farm record observations. Associations between KAP scores and farmer demographics were analyzed using multivariable logistic regression. Almost all farmers (98.3%) recognized antibiotics, but only 55% understood withdrawal periods, 58.3% recognized residues, and 35.8% knew these could transfer through milk or meat. Awareness of AMR was poor: 64.2% denied its link to dosage, and 52.9% were unaware of associated public health risks. Misuse was common: 68.7% used antibiotics without prescriptions, 69.2% ignored dosage guidelines, and 55.4% failed to complete treatment courses. Only 20% kept antibiotic records, and 31.1% had AMR-related training. Education and training strongly influenced KAP. Farmers with secondary education and AMR training had better knowledge (OR: 11.70, 95% CI: 2.54–54.04), while higher secondary education was associated with positive attitudes (OR: 25.28, 95% CI: 3.67–174.76) and improved practices (OR: 24.81, 95% CI: 4.45–138.25). Dairy farmers exhibited lower KAP than beef-fatteners, whereas mixed farmers showed superior knowledge (OR: 14.73, 95% CI: 2.85–76.36), attitudes (OR: 29.30, 95% CI: 2.19–39.07), and practices (OR: 8.00, 95% CI: 1.00-64.07). These findings reveal critical knowledge gaps, misconceptions, and extensive misuse of antibiotics. Targeted education, farmer training, and stricter regulations — especially for high-risk groups such as dairy farmers — are essential to promote responsible antibiotic use and mitigate AMR risks.</p>