Systematic Mapping of Ruminant Milk Extracellular Vesicles using Text Mining and Topic Analysis approach

Ruminant milk-borne extracellular vesicles (EVs) have garnered significant 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, metabolic and stress regulation. These distinctive characteristics of EVs position them as potential indicators of physiological and disease conditions in both human and animal research. Further, thermal stress induced alterations in the biological system of the animal are mirrored in these milk-borne molecular structures, indicating them to be accessible, non-invasive markers of heat stress in ruminant livestock. However, a holistic comprehension of the role of ruminant milk-derived EVs in human and livestock health remains limited. In spite of rapid developments in this field, 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 ruminant milk-derived EV's isolation, function, cargo profiling, biomarker prospects and potential applications. 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 the structured search using the keywords "extracellular vesicle" AND milk, combined with additional terms including "heat stress," "cell culture," "buffalo," "cattle," "sheep," and "goat." Descriptive statistics, along with text mining and topic analysis, were performed. Publications on ruminant milk-derived EVs began in 2012, with marked increase after 2021 and significant peak in 2024 with majority of publications in the International Journal of Molecular Sciences and Journal of Dairy Science. Major proportion of publications originated from 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 EVs in the context of biomedicine, nutraceuticals and human health compared with the diminished concentration on livestock research aspects like heat stress. The 7 identified topics following topic analysis spanned distinct subjects including ruminant milk EV isolation, characterization, functioning, immune-modulatory role and applications. The analysis also revealed significant challenges in method standardization, characterization protocols clinical validation and supportive regulatory frameworks. Research skew toward miRNA, overlooking other biomolecules was acknowledged; amidst EVs applications in livestock, especially under climate stress, remaining unventured. Further, key gaps in toxicity, safety and bioavailability was also identified demanding inter-disciplinary collaboration for diagnostic and therapeutic deployment across species. Bridging these gaps remains crucial for unlocking the maximum potential of ruminant milk-borne EVs in advancing the One Health framework and for the attainment of sustainable development goals.