Fermented milk is the optimal vehicle for delivering probiotic bacteria. However, the viable count of probiotic bacteria such as some lactic acid bacteria and the post-acidification of fermented milk are a contradiction. The objective of this study was to restrict the post-acidification of the fermented milk containing living Lactobacillus rhamnosus hsryfm 1301. Mild heat stress treatment (46°C, 1h) was chosen to help control the post-acidification caused by L. rhamnosus hsryfm 1301. When fermented milk was produced by single L. rhamnosus hsryfm 1301, the heat stress treatment reduced the post-acidification from 0.39 to 0.11% lactic acid, and the viable cells were maintained above 2.0 × 108CFUmL-1 during 21days of storage. Although the post-acidification limitation of heat treatment was relatively weak in fermented milk produced by L. rhamnosus hsryfm 1301 and S. thermophilus grx02 (from 0.26 to 0.10% lactic acid), this treatment was still effective. Furthermore, no whey separation in the fermented milk was caused by the treatment. Mild heat stress treatment could limit the post-acidification caused by L. rhamnosus hsryfm 1301 by decreasing its metabolism and proliferation. This treatment is a promising strategy to improve the shelf life of probiotic fermented milk.