Flow cytometric profiling of water buffalo milk EVs: how lactation stage drives phenotypic evolution and macrophage modulation.

@article{canonico2026flow,
  title = {{Flow cytometric profiling of water buffalo milk EVs: how lactation stage drives phenotypic evolution and macrophage modulation.}},
  author = {Barbara Canonico and Mariele Montanari and Ludovica Di Fabrizio and Giulia Franzoni and Giovanna De Matteis and Katia Cappelli and Francesco Grandoni and Maria Carmela Scatà and Daniele Lopez and Samanta Mecocci and Michela Battistelli and Tania Vanzolini and Giovanna Cappelli and Anna Donniacuo and Orlando Paciello and Caterina Ciacci and Stefano Papa and Michele Guescini and Paola Lanuti and Alessandra Martucciello},
  journal = {Frontiers in veterinary science},
  year = {2026},
  month = oct,
  doi = {10.3389/fvets.2026.1922591},
  url = {https://europepmc.org/article/MED/42904089},
  note = {CC BY},
}

TY  - JOUR
TI  - Flow cytometric profiling of water buffalo milk EVs: how lactation stage drives phenotypic evolution and macrophage modulation.
AU  - Barbara Canonico
AU  - Mariele Montanari
AU  - Ludovica Di Fabrizio
AU  - Giulia Franzoni
AU  - Giovanna De Matteis
AU  - Katia Cappelli
AU  - Francesco Grandoni
AU  - Maria Carmela Scatà
AU  - Daniele Lopez
AU  - Samanta Mecocci
AU  - Michela Battistelli
AU  - Tania Vanzolini
AU  - Giovanna Cappelli
AU  - Anna Donniacuo
AU  - Orlando Paciello
AU  - Caterina Ciacci
AU  - Stefano Papa
AU  - Michele Guescini
AU  - Paola Lanuti
AU  - Alessandra Martucciello
PY  - 2026
DA  - 2026/10/06
JO  - Frontiers in veterinary science
DO  - 10.3389/fvets.2026.1922591
UR  - https://europepmc.org/article/MED/42904089
AB  - Introduction  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 (Bubalus bubalis) 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' physical properties and their ability to modulate the immune response in macrophages.Material and methods  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.Results and conclusion  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's 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-1α 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.
SN  - 2297-1769
ER  -