Development of dairy products containing bifido bacteria is one of the main focuses in the food industry, considering the health benefits attributed to its survival through gut intestinal tract and the liberation of biogenic compounds. In order to evaluate the effect of process conditions on survival of Bifidobacterium animalis subsp. Lactis HN019 and release of biogenic compounds during cold storage two technological processes were employed: (i) addition of Probiotic (unfermented bifido milk; UFBM) and (ii) fermentation (fermented bifido milk; FBM). B. Lactis HN019 counts were stable only in fermented milks during seven weeks of cold storage. Moreover, dairy matrices associated with cold storage appeared to protect bifidobacteria during exposition to gastric conditions, assuring the correct Probiotic counts at gut entrance. The distribution of fatty acids was affected by the different Probiotic products. In contrast, fermentation process increased the relative contents of some bioactive fatty acids, such as elaidic acid (+11%) and conjugated linoleic acids (+20%). It was also noted an increase in polyunsaturated fatty acids in the FBM and a higher relative content of monounsaturated fatty acids in the UFBM, as compared to control milk. In addition, fermentation process by bifido bacteria enhanced the long chain fatty acids fraction in bifido milks as compared to control and unfermented bifido milks. Control milk and unfermented bifido milk showed the same peptides even after seven days of storage in contrast with fermented milks that had increase bioactive peptides. It is possible to suggest that opioid peptides are formed during fermentation process, increasing the source of bioactive peptides. Finally, the cold conservation process modified peptides and could improve antibacterial activity.