Swiss young generations prefer soft and fresh cheeses over semi-hard and hard varieties (Agristat, 2025). To produce these, cheesemakers rely on starter cultures- mixtures of defined or undefined bacterial strains - that drive milk acidification and develop specific aromas, flavors, and textures during relatively short ripening periods. The aim of this project was to isolate mesophilic lactic acid bacteria (LAB) from raw cow, goat, and sheep milk, in order to expand an existing strain collection and identify new isolates with desirable technological traits. A total of 91 milk samples were collected from various Swiss Cantons, sub-cultivated in milk, and initially selected based on acidification capacity and flavor. Identified strains included L. lactis subsp. lactis, L. cremoris, L. mesenteroides, S. thermophilus, L. raffinolactis, Lb. paracasei, and Lb. plantarum, as well as two wild species: Pseudolactococcus laudensis and L. taiwanensis. A high-throughput screening (HTS) assessed acidification speed, proteolytic activity, and inhibitory effects against L. innocua and S. aureus. Carbon source utilization profiles were analyzed on selected isolates using the Biolog™ PM01–PM02 assay. For L. cremoris, volatile compounds were identified through a semi-quantitative GC-MS analysis.Results revealed high variability among isolates, particularly in relation to sub-cultivation temperature and geographical origin. HTS proved effective in generating an overview of phenotypic profiles, enabling the selection of promising candidates for safety assessment and screening for transferable antibiotic resistance genes (ARGs). These steps represent the foundation for the development of novel starter cultures.