{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T14:50:17Z","timestamp":1791298217066,"version":"4.1.0"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Milk is more than a traditional nutrient source; it serves as a crucial carrier for milk-derived extracellular vesicles (mEVs), food-derived nanoparticles with diverse effects connecting animal health and human nutrition to ecological stability. The primary goal of this study was to characterize the phenotype and functional impact of mEVs from water buffalo (\n                      <jats:italic>Bubalus bubalis<\/jats:italic>\n                      ) across the lactation curve. The researchers aimed to determine how the transition from colostrum (Colo EVs) to early milk (EM-EVs) and mature milk (MM-EVs) alters the vesicles\u2019 physical properties and their ability to modulate the immune response in macrophages.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Material and methods<\/jats:title>\n                    <jats:p>The study utilized Flow Cytometry (FC) as a core tool, first validating specific antibody clones for use in the buffalo species. The functional impact was tested on both a bovine cell line (BoMac) and primary water buffalo macrophages, focusing on cellular uptake, mitochondrial oxidative stress (ROS) and phagocytic activity.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results and conclusion<\/jats:title>\n                    <jats:p>MM-EVs play the most important role in boosting phagocytosis in bubaline macrophages, accompanied by the highest increase in CD14 surface density, whereas Colo-EVs demonstrated the highest degree of internalization and the strongest antioxidant activity. In conclusion, the study demonstrates that mEVs provide a biological bridge during the calf\u2019s development. While Colo-EVs focus on antioxidant protection and systemic signaling, MM-EVs actively enhance the innate immune defense. This provides a mechanistic explanation for why continued consumption of maternal milk beyond the colostral phase is vital for the immunological resilience and survival of buffalo calves. Furthermore, BoMac cells and bubaline macrophages exhibit distinct cytokine profiles, as IL-1\u03b1 displays an increasing trend that did not reach statistical significance in MM-EV-treated BoMac cells, whereas a non-significant increasing trend is observed in macrophages treated by both Colo EVs and MM-EVs. Ultimately, the conserved nature of tetraspanins and miRNA profiles underscores the broad cross-species translational potential of buffalo mEVs, extending to human applications.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2026.1922591","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T13:50:22Z","timestamp":1791294622000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Flow cytometric profiling of water buffalo milk EVs: how lactation stage drives phenotypic evolution and macrophage modulation"],"prefix":"10.3389","volume":"13","author":[{"given":"Barbara","family":"Canonico","sequence":"first","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, 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Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniele","family":"Lopez","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samanta","family":"Mecocci","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Perugia","place":["Perugia, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michela","family":"Battistelli","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tania","family":"Vanzolini","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanna","family":"Cappelli","sequence":"additional","affiliation":[{"name":"National Reference Centre for Hygiene and Technology of Breeding and Buffalo Production, Istituto Zooprofilattico Sperimentale del Mezzogiorno","place":["Salerno, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Donniacuo","sequence":"additional","affiliation":[{"name":"National Reference Centre for Hygiene and Technology of Breeding and Buffalo Production, Istituto Zooprofilattico Sperimentale del Mezzogiorno","place":["Salerno, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Orlando","family":"Paciello","sequence":"additional","affiliation":[{"name":"National Reference Centre for Hygiene and Technology of Breeding and Buffalo Production, Istituto Zooprofilattico Sperimentale del Mezzogiorno","place":["Salerno, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caterina","family":"Ciacci","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefano","family":"Papa","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michele","family":"Guescini","sequence":"additional","affiliation":[{"name":"Department of Biomolecular Sciences, DISB, University of Urbino Carlo Bo","place":["Urbino, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Lanuti","sequence":"additional","affiliation":[{"name":"Department of Medicine and Aging Sciences, University \u201cG.D\u2019Annunzio\u201d Chieti-Pescara","place":["Chieti, Italy"]},{"name":"Center for Advanced Studies and Technology (CAST), University \u201cG.D\u2019Annunzio\u201d Chieti-Pescara","place":["Chieti, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Martucciello","sequence":"additional","affiliation":[{"name":"National Reference Centre for Hygiene and Technology of Breeding and Buffalo Production, Istituto Zooprofilattico Sperimentale del Mezzogiorno","place":["Salerno, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,10,6]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/vetsci10050358","article-title":"Colostrum composition, characteristics and Management for Buffalo Calves: a review","volume":"10","author":"Lotito","year":"2023","journal-title":"Vet Sci"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"806","DOI":"10.1186\/S12864-015-2044-9","article-title":"MicroRNA 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