ABSTRACT Background Bovine mastitis (BM) is a major source of morbidity among cattle, imposing a significant burden on the dairy industry. Understanding the molecular mechanisms that confer susceptibility to bovine mastitis offers potential for therapeutic targeting, but these mechanisms remain complex and incompletely understood. Objectives This study aims to identify key molecular players and regulatory networks involved in bovine mastitis by integrating multiple expression datasets and applying network analysis. Methods We integrated data from three BM expression datasets and applied weighted gene co‐expression network analysis (WGCNA) to identify specific modules and hub genes related to BM. Within the module that best correlated with mastitis, we constructed a network of mRNA–miRNA and protein–protein interactions. Results Our analysis revealed that TNFRSF1A and NCF1 are implicated as hub genes in the host's immune response to BM. Additionally, we identified bta‐mir‐2881 as a potential regulator of both TNFRSF1A and NCF1 . These genes are crucial in regulating the immune response and defence against bacterial infections, highlighting their importance in mastitis susceptibility. Conclusion Identifying miRNAs like bta‐mir‐2881 that may target key genes involved in mastitis susceptibility can shed light on the molecular mechanisms underlying the disease. This provides new targets for BM prevention and treatment, potentially alleviating the significant burden of the disease on the dairy industry.