The dairy sector is essential for the world's food systems, playing a vital role in agricultural production. Yet, mastitis, an inflammation of the mammary gland, affects a considerable portion of the dairy cow population annually, leading to reduced productivity and sustainability. Insight into therapeutic protocols is essential for a better understanding of the situation on farms with the aim of developing new or harmonizing existing protocols in the mastitis treatment. This study conducted on dairy farm in Serbia aimed to explore the most commonly used antibiotics during bovine mastitis treatment and their association with daily milk yield. Data from 100 lactating cows, including therapeutic protocols, lactation numbers, mastitis episodes, milk yield, and antibiotic usage, were obtained from a computerized database and statistically analysed. The results demonstrated a high prevalence of narrow‐spectrum antibiotics usage, with the majority of cows treated with combination therapy. The route of antibiotic application did not significantly impact daily milk yield, while treatment duration had no discernible effect. Nonetheless, specific antibiotics were associated with milk yield variations, suggesting the need for careful antibiotic selection and management in mastitis therapy. These findings emphasize the importance of prudent antibiotic use to safeguard animal health and long‐term milk production sustainability.