Donkey milk represents a valuable alternative for diversifying dairy resources. The mixture of donkey milk with milk from other species may help overcome its technological limitations while creating opportunities for the development of innovative cheeses based on non-conventional milk sources and indigenous lactic acid cultures. The effects of adjunct Lacticaseibacillus and ripening time on the physicochemical, microbiological, proteolytic, volatile, and sensory characteristics of cheese were evaluated. Cheeses made from a mixture of sheep milk (70%) and donkey milk (30%) were manufactured using a commercial starter culture and served as the control. Experimental cheeses were produced from the same milk blend inoculated with the commercial starter culture and supplemented with autochthonous Lacticaseibacillus strains. Their addition accelerated acidification and modulated the volatile profile, increasing alcohol and ester concentrations while reducing aldehydes and branched-chain fatty acids. These changes may have contributed to enhanced aroma intensity and complexity, together with increased hardness and friability of the cheeses during sensory evaluation. The use of indigenous Lacticaseibacillus cultures may contribute to the production of innovative mixed-milk cheeses, supporting the sustainable valorisation of donkey milk in dairy systems.

Plant-derived coagulants are increasingly explored as alternatives to animal rennet. This study provides the first evaluation of the coagulation kinetics and technological properties of Onopordum platylepis Murb. in ewe’s milk. Response surface methodology was applied to optimize temperature (30, 33, and 36 °C), pH (5.5, 6.0, and 6.5), and calcium chloride concentration (2, 4, and 6 mM). The optimal conditions for minimizing milk-clotting time were 36 °C, pH 5.5, and 6 mM CaCl₂. Under standardized activity (50 IMCU/L), the Rheological properties of gels produced by Onopordum platylepis were compared with Cynara cardunculus, Cynara humilis, animal rennet, a commercial plant coagulant, and fermentation-produced chymosin. Onopordum platylepis showed slower curd-firming rates than animal rennet and Cynara cardunculus, but similar behavior to Cynara humilis. Gels produced with Onopordum platylepis exhibited firmness comparable to commercial plant coagulants. The water-holding capacity was similar to other coagulants, though protein losses were higher for Onopordum platylepis, Cynara humilis, and Cynara cardunculus than animal rennet. Overall, Onopordum platylepis demonstrates potential for ewe’s milk cheese production, in which highly proteolytic coagulants are used.