The novelty of being handled during milking contributes to peripartum stress in dairy heifers with possi-bly adverse effects on their well-being and health. The present study investigated (1) heifers’ behaviouralresponses to handling over the course of four standardized handling sessions and (2) whether this han-dling just before parturition led to reduced heifers’ avoidance distances and reduced agitation behaviourin the milking parlour as well as improved udder health. The standardized handling was based on Telling-ton TTouch®and always started with touching the heifers’ hind quarters (HQ), proceeding over dorsum,neck, flank and belly (rest of the body) to hind legs and udder (HLU) as the target region. Altogether 13and 14 heifers on three farms were handled or served as control, respectively. Behavioural responses tothe handling in the four sessions were observed from videos in 10 of the handled heifers. All animalswere tested for their avoidance distance towards an experimenter before handling and 3 to 4 days p.p.Additionally, their agitation behaviour (stepping and kicking) during two milkings at 2 to 4 days p.p.,as well as somatic cell scores of the first three monthly milk test recordings (average and differencebetween first and third test day) were recorded and analysed with mixed models. In the four handlingsessions the relative duration of aversive responses to touching at HQ decreased after the first handling.Also the proportion of time in which touching of HLU was tolerated increased in the second and thirdhandling session compared to the first session. However, no significant differences between handled andcontrol heifers were found regarding their behaviour after parturition and their udder health, with largevariance in responses during milking. Possibly the handling was not intensive or long-lasting enough andagitation during the first milkings is not only related to fear of humans. Further research is needed tofind recommendable and time-efficient on-farm handling methods in order to reduce peripartum stressin heifers.