This study investigated the effect of stroking vs. simple human presence on later reactions of dairy cows to routine veterinary handling. While in two groups of cows the experimenter stroked the ventral part of the neck (Neck, N = 14) or the withers (Withers, N = 15) for three consecutive weeks, the third group was exposed to close visual presence (Control, N = 14). After the treatment period the cows were subjected to rectal palpation. The three groups differed significantly in stepping during rectal palpation, which occurred less often in Neck- and Withers-animals than in control animals. Heart rate increase was significantly higher in the control group than in the two stroking groups. Previous stroking led to fewer stress reactions during the rectal palpation, possibly due to a combined effect of improved relationship towards and thus perception of humans and lasting anti-stress effects of tactile stimulation.