Systematic mapping of ruminant milk extracellular vesicles using a text mining and topic analysis approach

Ruminant milk-borne extracellular vesicles (EV) have garnered substantial attention as assorted bioactive components of intercellular communication and key regulators of both physiological and stressful conditions. These membrane-bound vesicles transport diverse molecular cargo, contributing to immune modulation, cellular homeostasis, and metabolic and stress regulation. These distinctive characteristics of EV position them as potential indicators of physiological and disease conditions in both human and animal research. In addition, thermal stress-induced alterations in the biological system of the animal are mirrored in these milk-borne molecular structures, showing them to be accessible, noninvasive markers of heat stress in ruminant livestock. However, a holistic comprehension of the role of ruminant milk-derived EV in human and livestock health remains limited. In spite of rapid developments in this field, the variability in methodologies, investigations, and stated findings demands a comprehensive synthesis of existing knowledge. In this context, this work offers a systematic review of the aspects of the isolation, function, cargo profiling, biomarker prospects, and potential applications of ruminant milk-derived EV. Relevant articles were screened based on predefined inclusion criteria to identify significant research trends and thematic areas. A total of 124 studies published between 1990 and January 2026 were examined using Scopus data, following a structured search using the key words "extracellular vesicle" AND milk, combined with additional terms including "heat stress," "cell culture," "buffalo," "cattle," "sheep," and "goat." Descriptive statistics were calculated, and text mining and topic analysis were performed. Publications on ruminant milk-derived EV began in 2012, with a marked increase after 2021 and significant peak in 2024, with the majority of publications found in the International Journal of Molecular Sciences and Journal of Dairy Science. A major proportion of publications originated in China, followed by the United States, Australia, Italy, and Japan. The most frequent co-occurring terms were "extracellular vesicle," "milk," "exosomes," "bovine milk," and "drug delivery." Text mining results indicated strong research focus on ruminant milk-derived EV in the context of biomedicine, nutraceuticals, and human health compared with fewer studies on livestock research aspects such as heat stress. The 7 identified topics following topic analysis spanned distinct subjects including the isolation, characterization, functioning, immune-modulatory role, and applications of ruminant milk EV. The analysis also revealed substantial challenges in method standardization, characterization, protocols, clinical validation, and supportive regulatory frameworks. Research skewed toward microRNA, with less focus on other biomolecules, and EV applications in livestock, especially under climate stress, remaining unexplored. Further, key gaps in toxicity, safety, and bioavailability were also identified, demanding interdisciplinary collaboration for diagnostic and therapeutic deployment across species. Bridging these gaps remains crucial for unlocking the maximum potential of ruminant milk-borne EV in advancing the One Health framework and for the attainment of sustainable development goals.