Mycoplasma bovis is a globally significant pathogen of cattle associated with a wide range of clinical syndromes, including respiratory disease, mastitis, arthritis, otitis, and reproductive failure. Since its detection in Ireland in 1994, M. bovis has become a significant contributor to morbidity and mortality in Irish cattle. This study aimed to investigate herd-level risk factors associated with M. bovis seropositivity in replacement dairy heifers, using data from 105 Irish dairy herds. Ten heifers per herd were sampled on three occasions: spring 2018, spring 2019, and autumn 2019. Seropositivity was evaluated using two thresholds: ≥1 positive heifer (Model ≥ 1POS) and ≥3 positive heifers (Model ≥ 3POS). M. bovis seropositivity varied over time, with at least one positive heifer in 50.4% (95% confidence interval (CI): 40.5–60.3) of herds in spring 2018, 35.2% (95% CI: 26.2–45.1) in spring 2019, and 45.7% (95% CI: 36.0–55.7) in autumn 2019. Herds with three or more positive heifers increased from 31.4% (95%CI: 22.7–41.2) in spring 2018 to 42.9% (95% CI: 33.2–52.9) by autumn 2019. Risk factors for M. bovis seropositivity included the purchase of cattle, which significantly raised the odds of seropositivity across multiple visit periods (Model ≥ 1POS: Odds ratio (OR) 3.84, p = 0.02; Model ≥ 3POS: OR 3.69, p = 0.02). Managing more than three land parcels, housing heifer calves separately from bull calves, and sharing airspace between calves and older animals also increased seropositivity risks. Conversely, more colostrum feeds reduced the risk of seropositivity (Model ≥ 1POS: OR 0.81, p = 0.05), while colostrum quality assessment and feeding waste milk showed a trend toward increased risk. These findings suggest the importance of robust biosecurity measures, including limiting cattle purchases, improving calf management, and enhancing colostrum feeding practices, to control the spread of M. bovis. This study provides valuable insights into the epidemiology of M. bovis in Irish dairy herds, emphasising the need for targeted biosecurity and surveillance to safeguard herd productivity.
