¹Shihezi University, Faculty of Animal Science and Technology, Veterinary Medicine Department, 832000, CHINA
²Changji Hui Autonomous Prefecture Animal Disease Prevention and Control Center, 831100, CHINA
³Animal Husbandry and Veterinary General Station of Xinjiang Production and Construction Corps, 830063, CHINA
⁴Aheqi Animal Husbandry and Agriculture Development Co., Ltd, 843500, CHINA DOI : 10.9775/kvfd.2026.37459 Intercellular communication mediated by extracellular vesicles (EVs) is a key component of host-pathogen interactions. Emerging evidence suggests that Outer Membrane Vesicles (OMVs) enter host cells via multiple pathways and release cargo, including proteins and small RNAs (sRNAs), that regulate gene expression and function in recipient cells. In this study, we used Brucella melitensis 16M and its OMVs to construct an OMV-specific sRNA dataset and investigate their unique role in B. melitensis 16M intracellular survival. A total of 1,783,364 unique sRNA sequences were identified, and the ten most abundant sequences were determined by sequencing sRNAs extracted and purified from B. melitensis 16M and its OMVs. Bioinformatic analysis showed that eight sRNAs within OMVs exhibited stable secondary structures, and six were predicted to interact with mouse mRNAs. Notably, sRNA t0000835 was predicted to target the G protein-coupled receptor 45 (GPR45) gene and suppress its expression by binding to Argonaute 2 (AGO2). Transfection of sRNA t0000835 markedly enhanced intracellular survival of B. melitensis 16M. Collectively, these findings support the hypothesis that sRNAs t0000835 of B. melitensis 16M OMVs modulates host gene expression to promote the intracellular viability of B. melitensis 16M. Keywords : B. melitensis 16M, OMVs, Regulatory mechanisms, sRNA