Abstract
BACKGROUND: Inappropriate antimicrobial use (AMU) in livestock contributes to antimicrobial resistance (AMR), posing risks to both animal and human health. Despite growing awareness, antimicrobial stewardship (AMS) initiatives remain limited in Ethiopia’s dairy sector. This study evaluated the effects of farm-level AMS training on farmers’ knowledge and attitudes toward AMR and AMU, and disease incidence on dairy farms in Mekelle, Northern Ethiopia. METHODS: A prospective field intervention was conducted from December 2024 to June 2025 among 22 dairy farms with at least 20 lactating cows each. Farms were allocated to intervention (n = 11) and control (n = 11) groups based on geographical proximity. Baseline assessments preceded AMS training, with follow-up evaluations at three and six months. The AMS intervention included participatory training on prudent AMU, infection prevention, and record-keeping. Changes in farmers’ knowledge and attitudes were measured using structured questionnaires; AMU was quantified as treatments per 100 cow-days, and disease incidence as new cases per 100 cow-days. RESULTS: Following AMS training, mean knowledge scores in the intervention group increased from 48.2% to 78.6% (p < 0.05), while control farms did not significantly change (47.3% to 51.5%). Attitude scores changed from 3.1 to 4.2 (p < 0.05). Antimicrobial treatment rates changed from 4.5 to 2.0 per 100 cow-days in the intervention group (56% reduction), with minimal change in controls (4.1 to 3.9). Disease incidence declined from 7.9 to 4.5 cases per 100 cow-days, a 43% reduction, with the largest decreases observed for mastitis (52%) and respiratory disease (47%). CONCLUSION: Targeted AMS training was associated with change of farmers’ knowledge and attitudes, reduction of AMU, and decreased disease incidence on dairy farms. These findings may show the effect of participatory, farm-level stewardship programs in promoting responsible AMU and herd health, with potential benefits for One Health. Sustained follow-up and integration into veterinary extension services are recommended to maintain long-term impact. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12917-026-05385-z.