Abstract.To avoid the problems associated with twinning in dairy cattle, one of the embryos may be eliminated. This study compares the effect on pregnancy maintenance of two embryo reduction techniques, manual rupture (MR) and transvaginal ultrasound-guided aspiration (TUGA) of allanto-amniotic fluid, in Holstein-Friesian cows with multiple pregnancies. In the first experiment, 61 lactating cows bearing unilateral twins (n=27), bilateral twins (n=30) or triplets/quadruplets (n=4) were subjected to MR (n=45) or TUGA using a 17-G neddle (n=16) on day 28-34 of gestation. In 21 and 10 cows undergoing MR and TUGA embryo reduction, respectively, pregnancy loss occurred before day 90 (46.7 vs. 62.5%, P= 0.28). Through binary logistic regression, the type of pregnancy was identified as the only variable significantly affecting pregnancy maintenance (P=0.03). Based on the odds ratio, the risk of pregnancy loss was 4.1 times higher for unilateral twins than for bilateral twins (70.4 vs. 36.7%, respectively, P=0.01). No effect was detected on pregnancy maintenance of the technique used (P=0.17) or of the interaction technique by type of pregnancy (P=0.22). In the second experiment, a 22-G needle was used to perform TUGA on 22 lactating cows. The pregnancy loss rates were 44.4% (4/9), 18.2% (2/11) and 50% (1/2) for cows bearing unilateral twins, bilateral twins and triplets, respectively. The total pregnancy loss rate following TUGA using the 22-G needle tended to be lower than that using the 17-G needle (31.8 vs. 62.5%; P=0.06). Our results suggest that TUGA using a 22-G needle could be the method of choice to perform embryo reduction in cows carrying multiple pregnancies. M ultiple pregnancies (twins or higher-order pregnancies) are undesirable in dairy herds. Whilst cows carrying triplets and quadruplets are unlikely to maintain their pregnancies to term [1], cows delivering twins are more likely to suffer peripartum diseases and to be culled [2]. Twinning increases not only the risk of pregnancy loss [3,4] but also the risks of dystocia, stillbirth and calf mortality [5][6][7][8]. Over the past 20 years, twinning rates in dairy cattle have increased alongside milk production [2,5,9]. Estimates currently run at 9%, with rates of 0.3% to 12% reported among herds [10], and it is foreseeable that this twinning rate will continue to increase.Transforming a multiple pregnancy into a singleton pregnancy by inducing embryo reduction should in theory avoid the negative effects of multiple pregnancies in dairy cattle. However, the embryo reduction technique itself may carry an additional risk of pregnancy loss. Embryo reduction by manual rupture (MR) of the amniotic vesicle during the late embryonic/early fetal period has been assessed in unilateral [11,12] and bilateral twin pregnancies [12]. In these studies, pregnancy loss rates of 28.6 to 100% were reported. Transvaginal ultrasound-guided aspiration (TUGA) is an alternative method to MR for embryo reduction in twin-pregnant mares [13][14][15], although as far as we know, TUGA...