SUMMARY: Mammary secretion (1·2–2·0 ml) was collected from the milk sinus of each mammary gland (quarter) of two pregnant heifers and eight pregnant cows (dry period 49–229 days), first at weekly intervals from 40 days before parturition, then with increasing frequency as parturition approached. The progressive changes in the concentration of lactose, glucose, casein, non-casein protein and fat in the mammary secretion were determined. Calves were separated from the cows immediately after birth and the yield and composition of milk from individual quarters were determined for 5 days after parturition. Two quarters (milked quarters) of each of a further five cows were milked throughout pregnancy, while the other two quarters (unmilked quarters) of each cow were allowed to involute (dry off) 63– 104 days before parturition. Small samples (5–10 ml) of secretion were collected from the unmilked quarters, first at weekly intervals after drying off and then every second day from 10 to 15 days before parturition. On the days that the unmilked quarters were sampled, corresponding composite milk samples were collected from the milked quarters and the progressive changes in the yield of milk, lactose and fat were determined. The changes in the concentration of lactose and glucose in the mammary secretion, during drying off in late lactation, were determined in an additional five cows. In most cows allowed a usual dry period, and in the unmilked quarters of cows in which two quarters were milked throughout pregnancy, the concentration of lactose gradually increased from about 8 to 12 days before parturition to reach levels of about half those found in normal milk just before parturition. However, in some cows the concentration of lactose was low until 1–2 days before parturition and then increased abruptly, whereas in others the concentration of lactose increased slowly from as early as 32 days before parturition. A further rapid increase in the concentration of lactose in the mammary secretion occurred between 0 and 4 days before parturition in all cows. This latter increase was accompanied by an abrupt increase in the yield of milk, lactose and fat in the milked quarters of the cows in which two quarters were milked throughout pregnancy. These observations suggest that the initiation of lactation in the cow develops in two phases, a limited secretion of milk constituents occurs in late pregnancy and then 0–4 days before parturition copious secretion (lactogenesis) occurs.