Abstract
Milk represents not only a source of essential nutrients but also a versatile matrix for the development of functional foods when combined with lactic acid bacteria (LAB) fermentation. While cow milk is the most widely consumed, alternative matrices such as goat and donkey milk possess distinctive compositional features that may influence the bioactivity of fermented products. In this work, fifteen different strains belonging to LAB and bifidobacteria were initially tested for their ability to ferment cow, goat, and donkey milk. Fermented samples showing the best acidification rate were further screened for total phenolic content (TPC), radical scavenging activity (RSA), and effects in human colon epithelial cells and Caco-2 adenocarcinoma cells. The results showed that fermentation modulated TPC in a strain- and matrix-dependent manner, with donkey milk showing the most consistent increases. RSA was significantly enhanced in fermented samples, particularly those inoculated with Lacticaseibacillus casei BGP93 and LC4P1 strains and Limosilactobacillus reuteri ATCC23272. Cell viability assays confirmed the absence of cytotoxicity, and fermented milk extracts reduced ROS under induced oxidative stress. Antimicrobial assay showed that Lactiplantibacillus plantarum LPAL selectively inhibited Listeria monocytogenes, with the strongest effect observed in donkey milk. These findings suggest that LAB-fermented milks from goats and donkeys may represent safe functional foods with improved antioxidant and antimicrobial activities.