Synchronization protocols for fixed-time embryo transfer (ET) contribute significantly to the overall cost of an in vitro-produced-ET program, primarily through the cost of drugs and the labour required. Optimization of synchronization protocols to reduce cost, while providing high fertility, have the potential to improve overall efficiency and profitability. The objective of the present study was to evaluate the effect of dose and schedule of administration of prostaglandin F_2α (PGF) during a synchronization protocol for fixed-time ET. Holstein and cross-bred Holstein heifers (n = 3766) were synchronized using a modified 5-day CIDR Synch as follows: Day 0: CIDR inserted; Day 5: CIDR removed, PGF_2α treatment; Day 8: gonadotropin-releasing hormone (GnRH; 100 μg of gonadorelin). On Day 5, at the time of CIDR removal, heifers were randomly assigned to a 2 × 2 factorial design to receive either a full or half dose of PGF (Cloprostenol; 500 v. 250 μg) and 1 (Day 5) or 2 (Day 6) administrations resulting in the following treatments: full dose Day 5 + Day 6 (n = 938); full dose Day 5 (n = 938); half dose Day 5 + Day 6 (n = 946); and half dose Day 5 (n = 944). Heifers were evaluated by ultrasonography 5 days after GnRH to determine presence and size of the corpus luteum. Heifers with a corpus luteum received a fresh in vitro-produced embryo 7 ± 1 days after GnRH administration, and pregnancy was determined by ultrasonography 32 and 60 days after GnRH. Fertility data were analysed by logistic regression and included the fixed effects of dose, time, and their interaction. Fertility results are shown in Table 1. Utilisation rate (transferred/treated) was not affected by dose (P = 0.66), time (P = 0.19), or their interaction (P = 0.17). The percentage of heifers detected in oestrus was not affected by dose (P = 0.13), time (P = 0.72), or their interaction (P = 0.89). There were no significant differences between doses of PGF (P = 0.32), time (P = 0.71), or their interaction (P = 0.80) on pregnancies per ET on Day 32. Similarly, no differences were found on pregnancies per ET on Day 60 between doses (P = 0.35), time (P = 0.96), or their interaction (P = 0.89). In addition, pregnancy loss between Day 32 and 60 was not affected by dose (P = 0.76), time (P = 0.66), or their interaction (P = 0.54). In conclusion, the use of a half dose of PGF once on Day 5 resulted in comparable utilisation rate and fertility as the observed with 2 full dose applications 24 h apart. As a result, the overall cost of the fixed-time ET program can be reduced by eliminating the need for a second PGF treatment and by decreasing the dose without compromising fertility. Table 1. Utilisation rate, oestrus expression, pregnancies per ET (P/ET), and pregnancy loss in recipients receiving either a full or half dose of prostaglandin F_2α on Days (D) 5 and 6 or once on D 5

Dose	Time	Utilisation rate, % (no./no.)	Oestrus, % (no./no.)	P/ET D32, % (no./no.)	P/ET D60, % (no./no.)	Pregnancy loss, % (no./no.)
Full	D5 + D6	89.5 (839/938)	72.5 (680/938)	43.2 (362/838)	34.7 (291/838)	19.6 (71/362)
	D5	86.6 (812/938)	69.8 (655/938)	43.2 (351/812)	34.8 (283/812)	19.4 (68/351)
Half	D5 + D6	89.0 (842/946)	73.5 (695/946)	44.5 (375/842)	36.6 (308/842)	17.9 (67/375)
	D5	89.2 (842/944)	73.2 (691/944)	45.3 (381/842)	36.0 (303/842)	20.5 (78/381)