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The onset of lactation and the subsequent habituation to the milking routine is a stressful period, particularly for primiparous (PRI) cows. The objective of this study was to describe the dynamics of milking unit kick-off (KO) behavior in PRI cows during the first 3 mo of lactation, considering multiparous (MUL) cows as a reference for comparison. In addition, the potential associations between KO and milk yield and mastitis presentation were investigated. A total of 869 cows (PRI = 199; MUL = 670) on a dairy farm in northern CO were included in the analysis. Cows calving between August and November 2020 were enrolled from 3 DIM until 90 DIM. Participants were milked 3×/day in a 60-unit rotary parlor and data from each milking session were downloaded from parlor management software. Milking unit kick-off was used as a proxy for habituation to the milking procedure. Kick-off events were reported by the milking system and defined as an abrupt interruption in the milk flow during the milking process. Cow KO events were recorded for each milking session. Subsequently, occurrence of KO was analyzed by grouping the 3 consecutive milking sessions in each day and categorized as yes or no, indicating whether or not an individual kicked at least once in a given day. Data were analyzed by repeated measures ANOVA and logistic regression, including parity category, calving season, occurrence of dystocia, and their potential interactions in the models. Least squares means for daily proportions of KO were calculated considering the number of cows with KO events per day in the PRI and MUL categories. Subsequently, odds ratios for the occurrence of KO in PRI versus MUL were calculated at multiple periods of time. When KO was analyzed by DIM, proportions of KO were greater in PRI than in MUL during the whole monitoring period. In PRI, proportions of KO increased from 0.10/d to 0.20/d between 3 DIM and 15 DIM, to start decreasing around 30 DIM and remaining above MUL up to 90 DIM. On the contrary, in MUL cows, proportions of KO remained close to 0.05/d during the 90-d period. Overall, the odds of KO were greater for PRI versus MUL cows (odds ratio [95% confidence interval] = 2.07 [1.58-2.73]). No differences in milk yield were established among KO categories, while the percentage of cows affected with mastitis was greater in cows grouped in the quartile with more frequent KO events. We concluded that the relationship between days in milk and the proportion of PRI cows displaying KO was not linear, but rather KO increased during the first 2 wk postpartum before decreasing after the first month of lactation. First-parity cows had greater levels of KO than MUL cows, which is most likely associated with the process of habituation to milking during their first lactation.