Use of embryo transfer seven days after artificial insemination or transferring identical demi-embryos to increase twinning in beef cattle.

Our objectives were to determine pregnancy rate, fetal loss, and number of calves born in beef cattle after a fixed-time transfer of an embryo 7 d after a fixed-time artificial insemination (TAI) of cows (Exp. 1) and after transfer of 2 demi-embryos into a single heifer recipient (Exp. 2). In Exp. 1 after synchronization of ovulation, during 2 yr, 297 suckled beef cows were assigned randomly to 1 of 3 treatments: 1) on d 2 cows received a single TAI (TAI-2; n = 99), 2) a fixed-time direct transfer, frozen and thawed embryo placed in the uterine horn ipsilateral to the ovary containing a corpus luteum (CL) on d 9 embryo transfer (ET-9; n = 99), or 3) cows received TAI on d 2 and a frozen and thawed direct transfer embryo placed in the uterine horn ipsilateral to the ovary containing a CL on d 9 (TWIN) treatments (n = 99). Fetal number and viability were determined with ultrasonography at 33 to 35 d and 90 to 100 d after insemination. In Exp. 2, 74 crossbred recipient heifers were assigned randomly to receive either 1) a single whole fresh embryo (WHOLE; n = 37) or 2) 2 identical fresh demi-embryos (SPLIT; n = 37) in the uterine horn ipsilateral to the CL 7 d after an observed estrus. Ultrasonography was used on d 33, 69, and 108 to determine presence and number of embryos or fetuses. Palpation per rectum was used to determine pregnancy status on d 180 of gestation and number of live calves was recorded at birth. In Exp. 1 pregnancy rates on d 30 to 35 were greater (P < 0.05) for TWIN- (48.5%) and TAI-2- (47.5%) than for ET-9- (33.3%) treated cows. Of the 48 pregnant cows in the TWIN treatment, 21 were twin pregnancies whereas there was 1 twin pregnancy in the TAI-2 treatment. As a result, TWIN cows had more fetuses (P < 0.05) as a proportion of all treated cows (69.7%) than TAI-2- (48.5%) or ET-9-(33.3%) treated cows, and cows in the TWIN treatment gave birth to more (P < 0.01) calves (n = 55) compared with cows in the ET treatment (n = 23) whereas cows in the TAI-2 treatment (n = 40) were intermediate. In Exp. 2 heifers receiving SPLIT (81.1%) had greater (P < 0.05) pregnancy rates on d 33 than heifers receiving WHOLE (40.5%). Of the SPLIT heifers that were confirmed pregnant at d 33 after transfer, 57% were gestating twin fetuses. Embryonic or fetal loss from d 33 to birth was greater (P < 0.01) in heifers in the SPLIT treatment (40.0%) compared with the WHOLE treatment (0.0%), but number of calves per female treated was greater (P < 0.05) in heifers in the SPLIT treatment (75.0%) compared with heifers in the WHOLE treatment (40.5%). We conclude that transferring an embryo into a cow 7 d after TAI did not increase the pregnancy rate in Exp.1. However, transferring 2 demi-embryos into a single heifer recipient increased pregnancy rate at 33 d of gestation whereas both methods of inducing twinning resulted in a greater number of calves per female treated. In addition, embryonic or fetal loss associated with unilateral twin pregnancies in heifers occurred at rates greater than those associated with single-fetus pregnancies.

Administration of human chorionic gonadotropin at embryo transfer induced ovulation of a first wave dominant follicle, and increased progesterone and transfer pregnancy rates.

We hypothesized that administration of hCG to recipients at embryo transfer (ET) would induce accessory CL, increase serum progesterone concentrations, and reduce early embryonic loss (as measured by increased transfer pregnancy rates). At three locations, purebred and crossbred Angus, Simmental, and Hereford recipients (n = 719) were assigned alternately to receive i.m. 1,000 IU hCG or 1 mL saline (control) at ET. Fresh or frozen-thawed embryos were transferred to recipients with a palpable CL on Days 5.5 to 8.5 (median = Day 7) of the cycle (Locations 1 and 2), or on Day 7 after timed ovulation (Location 3). Pregnancy diagnoses (transrectal ultrasonography) were done 28 to 39 d (median = 35 d) and reconfirmed 58 to 77 d (median = 67 d) post-estrus. At Location 1 (n = 108), ovaries were examined at pregnancy diagnosis to enumerate CL. More (P < 0.001) pregnant hCG-treated cows (69.0%) had multiple CL than pregnant controls (0%). Serum progesterone (ng/mL) determined at Locations 1 and 2 (n = 471) at both pregnancy diagnoses in pregnant cows was greater (P ≤ 0.05) after hCG treatment than in controls (first: 8.1 ± 0.9 vs 6.1 ± 0.8; second: 8.8 ± 0.9 vs 6.6 ± 0.7), respectively. Unadjusted pregnancy rates at the first diagnosis were 61.8 and 53.9% for hCG and controls. At the second diagnosis, pregnancy rates were 58.6 and 51.3%, respectively. Treatment (P = 0.026), embryo type (P = 0.016), and BCS (P = 0.074) affected transfer pregnancy rates. Based on odds ratios, greater pregnancy rates occurred in recipients receiving hCG, a fresh embryo (66.3 vs 55.5%), and having BCS >5 (62.3 vs 55.3%). We concluded that giving hCG at ET increased incidence of accessory CL, serum progesterone in pregnant recipients, and transfer pregnancy rates. Furthermore, we inferred that increased progesterone resulting from hCG-induced ovulation reduced early embryonic losses after transfer of embryos to recipients.

Evaluating recipient and embryo factors that affect pregnancy rates of embryo transfer in beef cattle.

The objectives of this experiment were to determine the effects of corpus luteum characteristics, progesterone concentration, donor-recipient synchrony, embryo quality, type, and developmental stage on pregnancy rates after embryo transfer. We synchronized 763 potential recipients for estrus using one of two synchronization protocols: two doses of PGF2alpha (25 mg i.m.) given 11 d apart (Location 1); and, a single norgestomet implant for 7 d with one dose of PGF2alpha (25 mg i.m.) 24 h before implant removal (Location 2). At embryo transfer, ovaries were examined by rectal palpation and ultrasonography. Of the 526 recipients presented for embryo transfer, 122 received a fresh embryo and 326 received a frozen embryo. Pregnancy rates were greater (P < 0.05) with fresh embryos (83%) than frozen-thawed embryos (69%). Pregnancy rates were not affected by embryo grade, embryo stage, donor-recipient synchrony, or the palpated integrity of the CL. Corpus luteum diameter and luteal tissue volume increased as days post-estrus for the recipients increased. However, pregnancy rates did not differ among recipients receiving embryos 6.5 to 8.5 days after estrus (P > 0.1). There was a significant, positive simple correlation between CL diameter or luteal tissue volume and plasma progesterone concentration (r = 0.15, P < 0.01 and r = 0.18, P < 0.01, respectively). There were no significant differences in mean CL diameter, luteal volume or plasma progesterone concentration among recipients that did or did not become pregnant after embryo transfer. We conclude that suitability of a potential embryo transfer recipient is determined by observed estrus and a palpable corpus luteum, regardless of size or quality.

Effect of molybdenum-induced copper deficiency on in vivo and in vitro measures of neutrophil chemotaxis both before and following an inflammatory stressor.

Twelve Angus x Hereford heifers (avg wt = 183.6 kg) were allotted by initial liver copper (Cu) concentrations into one of two treatments. Control (n = 6) heifers were fed a basal diet supplemented to provide a dietary Cu level of 10 ppm. Molybdenum (Mo)-induced Cu-deficient heifers (n = 6) were fed an identical basal diet supplemented with sodium molybdate (Cu:Mo ratio = 1:2.5), with dietary sulfur at .3% of the total diet. Dietary treatments were delivered for 120 d, at which time Mo-supplemented heifers were considered Cu-deficient (286 and 49 ppm liver Cu for control and Mo-induced Cu-deficient, respectively). Peripheral blood neutrophils were enumerated both before and after the administration of an inflammatory stressor, a subcutaneous injection (1.5 mL) of Freund's complete adjuvant. In vitro and in vivo measures of neutrophil chemotaxis were evaluated and the expression of two adhesion molecules, CD18 and L-selectin, were analyzed by flow cytometric procedures. Molybdenum-induced Cu deficiency increased (P < .01) the number of peripheral blood neutrophils; however, in vitro neutrophil chemotaxis was not affected. In vivo neutrophil chemotaxis tended (P < .08) to be increased in Mo-induced Cu-deficient heifers (1.55 vs 2.26 x 10(6) cells/ sponge for control and Mo-supplemented, respectively). No differences in CD18 or L-selectin expression were detected between treatments. However, CD18 expression was decreased (P < .05) in both treatments following adjuvant injection. These data suggest that Mo-induced Cu deficiency results in an increase in peripheral blood neutrophil number, without altering chemotactic ability and adhesion molecule expression.

Acid-base status of a pulmonate land snail (Helix aspersa) and a prosobranch amphibious snail (Pomacea bridgesi) during dormancy.

Changes in metabolism and acid-base status were compared during dormancy in the pulmonate land snail Helix aspersa and a prosobranch amphibious snail Pomacea bridgesi. The typical condition of higher blood PCO2 and bicarbonate levels for air-breathing versus water-breathing vertebrates was shown for the two snail species. When exposed to dry air for 24 hr, both species depressed oxygen uptake by about 65%. In Pomacea, hypercapnia (increase in hemolymph PCO2 from 5.5 to 18 torr) resulting from dormancy produced no significant change in pH due to large increases in bicarbonate (over 17 mmol/l). In Helix, on the other hand, hypercapnia (increase in hemolymph PCO2 from 13 to 18 torr) resulting from dormancy produced a significant decrease in pH and a less than 7 mmol/l increase in bicarbonate. Pre-existing high levels of bicarbonate in Helix may prevent compensation of hypercapnia resulting from dormancy, similar to the case described for air-breathing vertebrates. Complete compensation of respiratory acidosis during the first 24 hr of dormancy in Pomacea suggests that metabolic rate suppression is independent of pH.