The objectives of this observational study were (1) to assess the time from the appearance of the amniotic sac (AS) or feet outside the vulva to birth in Holstein cows (primiparous and multiparous) with (dystocia) or without assistance (eutocia) at calving, and (2) to estimate reference times to be used as guidelines for obstetric intervention in Holstein cows that need assistance during difficult births. Cows (n=92) from 1 commercial dairy operation were used in this study. Periparturient dairy cows (primiparous, n=58; multiparous, n=34) were placed in a maternity pen and constantly monitored until birth. The calving ease of cows, time from AS or feet appearance to birth, calving progress from a subset of 15 cows (frequency and duration of abdominal contractions during labor), calf birth weight, calf sex, and stillbirths (born dead or died within 24h after birth) were recorded. The reference times for obstetric intervention during dystocia were estimated based on values from unassisted births (normal). The normal range of times from the appearance of AS or feet outside the vulva to birth was estimated based on the mean+2 standard deviations (SD) of unassisted births. According to farm protocol, assistance was provided to cows without calving progress 80 min after AS appearance or earlier (e.g., to correct malpositions). Cows with dystocic births had a longer time from AS appearance to birth and increased incidence of stillbirth compared with cows with eutocic calvings. After the appearance of the AS, calving progress was evident every 15 min for eutocic births. The estimated reference times (mean+2 SD) from AS appearance to birth were 69.7 min and from feet appearance to birth were 64.6 min for eutocic births. Findings from this study suggested that calving personnel should start assisting cows 70 min after AS appearance (or 65 min after feet appearance) outside the vulva. The time spent in labor (straining) combined with the time from the appearance of the AS or feet to birth, and the assessment of calving progress (as described for eutocic births) should be used as guidelines for obstetric intervention during difficult births under field conditions. These reference times should be interpreted in combination with adequate obstetrical knowledge and examination.