This study aims to identify spermatozoa kinematics and seminal plasma proteomic profiles as biomarkers of motility sperms in Bali-polled bull. Semen was collected using an artificial vagina twice a week until 5 batches. Kinematic sperms were analyzed using CASA and fertility was assessed by S/C of AI. Proteomic profiling of seminal plasma was confirmed using LCMS/MS. All the data were processed using the Uniprot protein database. Gene Ontology annotation determined the protein functions related to molecular function, biological processes, and cellular components, using Panther. Protein interactions were analyzed using STRING. The results showed that post-thawing kinematic sperm of Bali-polled bulls was considered a good category, with satisfactory fertility rate (S/C ratio of 1.52). The result of profile protein showed that the seminal plasma of Bali-polled bull showed a protein molecular weight distribution of 13 to 70 kDa identifying 138 proteins, 6 of which were found associated with sperm motility, including TEX101, BSP1, PRSS55, BSP3, SPADH2, and TPPP2. These proteins could serve as fertility biomarkers, providing additional parameters for bull selection. The study suggests that examining sperm kinematics and seminal plasma proteins in Bali-polled bulls could illuminate mechanisms controlling bull fertility and potentially lead to the development of superior local breeds.