The results of the presented study highlighted that the processing conditions such as the SCT and the IFpH altered the physicochemical and organoleptic properties of buffalo milk yogurt samples containing 3 g/100 g of milk fat. The IFpH mainly affected the activities of the strains in the mixed starter cultures. The probiotic cultures prolonged the fermentation time and decreased the reaction rate constant of postacidification during cold storage. Moreover, the extent of decrease in reaction rate constant varied depending on the IFpH. The absence of L. delbrueckii subsp. bulgaricus in the starter culture increased the viscosity for IFpH of 4.80. It also increased the syneresis for both IFpH 4.80 and 4.55. The IFpH did not affect the viscosity of T1 and T2, while it was vice versa for the other samples. Twenty days cold storage decreased the syneresis of the buffalo milk yogurt samples. L*, a*, and b* values of all the buffalo milk yogurt samples were not significantly affected by the IFpH and the SCT. The sensory results showed that a more acceptable buffalo milk yogurt was probiotic ABY‐2 culture added to yogurt samples. The organic acid and fatty acid profiles of the buffalo milk yogurt samples containing ABY‐2 culture were further analyzed and compared to the control yogurt containing YC‐X11 culture. The SCT significantly affected the production of lactic acid, acetic acid, and citric acid during cold storage. However, there was no significant effect of the SCT on the proportions of saturated and unsaturated fatty acids in the buffalo milk yogurt samples.