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We have previously shown that the addition of eCG improved pregnancy rates following fixed-time artificial insemination (FTAI) in lactating dairy cows treated with estradiol benzoate (EB) and progesterone releasing devices (Veneranda et al. 2006 Reprod. Fertil. Dev. 18, 118). An experiment was designed to compare pregnancy rates in lactating dairy cows treated with progesterone-releasing devices and EB plus eCG with different gonadotropin-releasing hormone (GnRH)-based protocols. Five-hundred lactating dairy cows between 30 and 51 days postpartum (when prostaglandin F (PGF) treatments were given in the Pre-Synch groups), with a milk yield of 29.5 � 7.0 kg per day (range 13.0 to 46.0 kg) and a body condition score (BCS) between 2.5 to 3.5 out of 5 were used. Cows were blocked by days postpartum and randomly assigned to five treatment groups. Cows in the Pre-Synch group received PGF 28 and 14 days prior to the insertion of a DIB device (1 g progesterone, Syntex SA, Buenos Aires, Argentina) and the administration of 50 µg of Lecirelin (GnRH, Ovusin, Syntex SA) IM (Day 0). On Day 7, DIB were removed and cows received PGF (0.5 mg cloprostenol, Ciclase, Syntex SA). On Day 9, cows received a second GnRH treatment and were FTAI 16 h later (60 h after device removal). Cows in the Modified Pre-Synch group were treated similarly except that they received a DIB for 7 days prior to the second PGF injection and no DIB at the time of the first GnRH. Cows in the P4-Synch group were treated as those in the Pre-synch group but did not receive the two doses of PGF 28 and 14 days earlier. Cows in the P4 + EB + eCG groups received a DIB for 8 days, 2 mg of EB at DIB insertion (Day 0), and PGF and 400 IU eCG (Novormon, Syntex SA) IM at DIB removal (Day 8); they were subdivided to receive 1 mg of EB on Day 9 (P4 + EB + eCG + EB group) or GnRH on Day 10 (P4 + EB + eCG + GnRH group). Cows in these latter two groups were also FTAI 60 h after device removal. Pregnancy was determined by rectal palpation 50 days after FTAI and data were analyzed by logistic regression. Pregnancy rates were not affected by BCS, days postpartum, or milk yield (P > 0.5). However, there was a significant group effect due to a higher (P < 0.05) pregnancy rate in the P4 + EB + eCG + EB (49%) and P4 + EB + eCG + GnRH (44%) treatment groups than in those in the Modified Pre-Synch group (30%). Pregnancy rate in the P4-Synch group (39%) was lower (P < 0.05) than those in the P4 + EB + eCG + EB group but not different from the others. Finally, pregnancy rate in Pre-Synch group (41%) was intermediate and not different from the other treatment groups. Results of these experiments suggest that treatments with progesterone-releasing devices, EB, and eCG result in pregnancy rates that are comparable to or better than those obtained in GnRH-based protocols in lactating dairy cows. Further studies are needed to investigate whether eCG could increase pregnancy rates in cows treated with GnRH and progesterone-releasing devices. The authors thank Syntex SA for providing the hormones used in the study.

In Holstein cows, the diameter of the dominant follicle (DF) at the time of follicle deviation is 8.5 mm and the subordinate follicle (SF) is 7.2 mm (Ginther et al. 1996 Biol. Reprod. 55, 1187–1194). However, follicular responsiveness to an ovulatory treatment occurs only with 10.0-mm-diameter follicles (Sartori et al. 2001 Biol. Reprod. 65, 1403–1409). The current study tested the hypothesis that, in Bos indicus (Nelore and crossbred Nelore � Gir) females, the follicular diameters at the time of deviation and ovulation responsiveness are smaller than those in Holstein cows. The experiment was performed in two phases. In the first phase, 12 Nelore heifers were previously synchronized with a protocol using progestagen and estradiol benzoate. After implant removal, all heifers were evaluated by transrectal ultrasonography (Aloka SSD-500, Tokyo, Japan) every 12 h until Day 5 of the estrous cycle (Day 0 = Day of the ovulation) to assess the time of ovulation, the time of follicle deviation, and the follicular diameter at the deviation. In the second phase, 29 Bos indicus heifers (Nelore and crossbred Nelore � Gir) were previously synchronized with the same protocol as cited above. After the ovulations (Day 0), the follicles were evaluated by transrectal ultrasonography every 24 h, until they reached the diameter of 7.0–8.4 mm (n = 9); 8.5–10.0 mm (n = 10); and >10.0 mm (n = 10). In order to assess the ovulatory capacity, all animals were treated with 25 mg of LH (Lutropin-V�; Bioniche Animal Health, Inc., Belleville, Ontario, Canada) at these follicle diameter ranges. After the LH treatment, all animals were monitored by ultrasonography every 12 h for 48 h. ANOVA, Bartlett, and chi-square tests were used in the statistical analyses. In the first phase, the diameters of the DF and SF at the time of follicular deviation (61.9 � 4.9 h after ovulation) were 6.2 � 0.2 and 5.8 � 0.2 mm, respectively. In the second phase, the the average follicular diameters at the time of LH administration in the groups 7.0–8.4 mm, 8.5–10.0 mm, and >10.0 mm were 7.6a � 0.1 mm, 9.6b � 0.1 mm, and 10.9c � 0.2 mm; and their ovulation rates were 33.3%a (3/9), 80.0%b (8/10), and 90.0%b (9/10), respectively (P < 0.05). The interval from LH treatment to ovulation was 38.0 � 4.0 h, 31.5 � 2.7 h, and 30.0 � 2.0 h, respectively (P > 0.05). In conclusion, in Bos indicus heifers, follicle deviation occurred with smaller diameters than previously reported in Bos taurus breeds. In addition, Bos indicus heifers are able to ovulate in response to 25 mg of LH with smaller diameters compared to those of Bos taurus breeds. Moreover, in Bos indicus heifers, ovulatory capacity is acquired by follicles as small as 7.0–8.4 mm, but this responsiveness significantly increases after follicles reach 8.5–10.0 mm. This work was supported by FAPESP (Proc:03/10203-4); Bioniche Animal Health, Inc., Belleville, Ontario, Canada; and Tecnopec, S�o Paulo, Brazil.

An experiment was designed to evaluate alternative treatment protocols for fixed-time embryo transfer (FTET) in recipients in order to reduce cattle handling. Non-lactating, cycling Bos taurus � Bos indicus beef cows (n = 918), with a body condition score (BCS) of 2.5 to 3.5 out of 5, received a progesterone-releasing device (DIB; Syntex, Buenos Aires, Argentina) and 2 mg estradiol benzoate (EB; Syntex) IM on random days of the cycle (Day 0); DIB were removed on Day 8. Cows were placed randomly into 6 treatment groups in a 3 � 2 factorial design. The factors evaluated were time of administration of PGF and eCG [control treatment: PGF Day 5 and eCG Day 5 (Bo et al. 2002 Theriogenology 57, 53–72); PGF Days 0 and 8 and eCG Day 5; PGF Days 0 and 8 and eCG Day 8] and the time of administration of the second injection of EB (at DIB removal or 24 h later). Therefore, cows in the control treatment received 150 �g of d-Cloprostenol (PGF, Ciclase; Syntex) and 400 IU eCG (Novormon; Syntex) on Day 5, whereas cows in the other 2 treatments received 75 �g of PGF on Days 0 and 8 and 400 IU eCG on either Day 5 or Day 8. Each treatment group was further subdivided to receive 1 mg EB IM on either Day 8 (EB Day 8) or Day 9 (EB Day 9). All recipients, whether observed in estrus or not and with more than one CL with luteal area (total CL area minus cavity area) >76 mm2 on the day before embryo transfer, received frozen-thawed embryos (IETS Grade 1) on Day 16 (EB Day 8) or Day 17 (EB Day 9) by direct transfer. Pregnancy was determined by ultrasonography 30 days after FTET. Continuous data were analyzed by ANOVA and pregnancy rates were analyzed by logistic regression. No effects of body condition score or technician were detected (P > 0.1). Although there were no differences in conception rates between recipients treated with EB on Day 8 or Day 9 (168/321, 52.3% vs. 203/350, 58.0%, respectively; P > 0.1), treatment with EB on Day 8 resulted in fewer recipients selected/treated (321/460, 69.7% vs. 350/458, 76.4%; P < 0.05) and pregnant/treated (168/460, 36.5% vs. 203/458, 44.3%; P < 0.05) than 24 h later (EB Day 9), respectively. The mean interval from DIB removal to estrus was shorter (P < 0.05) for recipients in the EB Day 8 (28.1 � 7.2 h) than in the EB Day 9 (43.5 � 9.4 h) group, and the mean (� SEM) CL area was also less in the EB Day 8 group than in the EB Day 9 group (261.7 � 96.7 vs. 293.1 � 114. mm2; P < 0.05). The number of recipients selected/treated was higher in the group receiving PGF on Days 0 and 8 and eCG on Day 8 (243/305, 79.6%) than in the group receiving PGF on Days 0 and 8 and eCG on Day 5 (208/301, 69.1%) or the control group (220/312, 70.5%). Although the luteal area was greater (P < 0.05) in the PGF on Days 0 and 8 and eCG on Day 5 group (304.4 � 127.1 mm2) than in the groups receiving PGF and eCG on Day 5 (274.6 � 98.6 mm2) or PGF on Days 0 and 8 and eCG Day 8 (258.4 � 90.2 mm2), pregnancy rates did not differ between PGF and eCG treatment groups (P > 0.05). It was concluded that it may be possible to reduce animal handling by treating recipients with a half dose of PGF at the time of DIB insertion and removal, when eCG is also administered, without compromising pregnancy rates.

An experiment was designed to evaluate the effect of the interval between thawing to deposition of the embryo into the uterine horn on pregnancy rates of in vivo-produced frozen–thawed embryos in 1.5 M ethylene glycol (direct transfer). Data were collected from 1122 embryo transfers performed in the same farm (Estancia El Mangrullo, Lavalle, Santiago del Estero, Argentina) during the spring and summer of 2004/05 and 2005/06 (6 replicates, ambient temperature between 20 and 40�C). Recipients used in all replicates were non-lactating, cycling, multiparous Bos taurus � Bos indicus crossbred cows with body condition score between 3 and 4 (1 to 5 scale) that were synchronized using fixed-time embryo transfer protocols. Briefly, the synchronization treatments consisted of the insertion of a Crestar ear implant (Intervet, Sao Paulo, Brazil) or a progesterone-releasing device (DIB; Syntex SA, Buenos Aires, Argentina), plus 2 mg of estradiol benzoate (EB; Syntex) intramuscularly (IM) on Day 0, and 400 IU of eCG (Folligon 5000; Intervet, or Novormon 5000; Syntex) IM plus 150 �g d-cloprostenol IM (Preloban; Intervet, or Ciclase; Syntex) on Day 5. Progestin devices were removed on Day 8 and all cows received 1 mg of EB IM on Day 9. All cows were examined by ultrasonography on Day 16 and those with a luteal area >76 mm2 (by calculating the area of the CL minus the area of the cavity) received, on Day 17, frozen–thawed embryos by nonsurgical transfer. All embryos were Grade 1, and all were frozen in 1.5 M ethylene glycol at the Embryo Plus Laboratory (Brits, South Africa). After being stored in liquid nitrogen, the embryos were plunged directly (no air thawing) in a 30�C water bath for 30 s, and then transferred to the recipient cows by either one of two technicians. Based on the interval between thawing and transfer, the transfers were classified as being in one of 3 groups: Group 1: 0.6. These results may be interpreted to suggest that there is no significant effect of time from thawing to transfer (up to 16 min) in direct transfer embryos using Bos taurus � Bos indicus recipients transferred at a fixed time.

In embryo transfer programs, 2 inseminations per superstimulated donor are usually recommended. Based on studies of follicular dynamics in Bos indicus donors (Baruselli et al. 2006 Theriogenology 65, 77–88), we tested the hypothesis that a single fixed-time AI (FTAI) at 16 h after pituitary luteinizing hormone (pLH) treatment is as efficacious as 2 FTAI at 12 and 24 h after the pLH treatment. Suckling Nelore cows (n = 10), 60 � 12 days postpartum, were equally distributed in 2 experimental groups: G-1TAI vs. G-2TAI, in a crossover design. All animals received an intravaginal progesterone device (DIB; Syntex S.A., Buenos Aires, Argentina) and 2 mg of estradiol benzoate (IM, Ric Be; Syntex) at random stages of the estrous cycle (Day 0). Superstimulatory treatments started on Day 4 with pFSH (133 mg; Folltropin-V; Bioniche Animal Health Canada, Inc., Belleville, Ontario, Canada) in 8 decreasing doses, given IM 12 h apart. On Day 6, all cows received 2 (AM and PM) treatments of 0.15 mg of d-cloprostenol (Prolise; Syntex). The DIB devices were removed 36 h after the first PGF2� injection. The pLH treatment (25 mg IM; Lutropin-V; Bioniche) was given 48 h after the first PGF2� treatment (12 h after the last pFSH treatment). In the G-2TAI group, all cows received 2 inseminations (12 and 24 h after the pLH). Cows in the G-1TAI group received a single insemination (16 h after the pLH). All inseminations were done with the same batch of semen from a single bull. Ultrasound examinations (PIE Medical Scanner 200; Pie Medical Equipment, Maastricht, The Netherlands) were performed every 12 h from 0 to 72 h after pLH treatment to evaluate the number of ovulatory follicles (8 mm at the time of pLH administration (16.2 � 1.4 vs. 14.8 � 1.2; P > 0.05), ovulation rate (63.8 � 3.8% vs. 64.2 � 4.3%; P > 0.05), interval from the first to the last ovulation (32.4 � 1.8 vs. 33.6 � 1.6 h; P > 0.05), total ova/embryos (8.2 � 0.9 vs. 7.2 � 0.8; P > 0.05), Grade 1 embryos (2.0 � 0.5 vs. 2.3 � 0.4; P > 0.05), transferable embryos (Grades 1, 2, and 3) (4.3 � 0.7 vs. 4.2 � 0.6; P > 0.05), freezable embryos (Grades 1 and 2) (2.9 � 0.6 vs. 2.8 � 0.4; P > 0.05), unfertilized ova (0.6 � 0.2 vs. 0.8 � 0.2; P > 0.05), and degenerated embryos (3.3 � 0.9 vs. 2.2 � 0.3; P > 0.05). These results suggest no significant differences in embryo production between superstimulated Nelore donors inseminated once or twice by fixed time following administration of pLH. This work was supported by Tecnopec, Brazil.

It has been shown than it is possible to AI embryo donors at a fixed time (FTAI) by removing the progestin-releasing device at the time of the last FSH treatment and inducing ovulation with gonadotropin-releasing hormone (GnRH) or porcine luteinizing hormone (pLH) at either 12 h (dairy cows: Bos indicus) or 24 h (beef cows: Bos taurus) after device removal (Bo et al. 2006 Theriogenology 65, 89–101). An experiment was designed to determine the best interval from device removal to GnRH or pLH treatment in Angus (beef cow) donors. Red Angus donors (43 cows and 56 heifers) were blocked by parity and randomly assigned to 1 of 4 treatment groups in a 2 × 2 factorial design. On Day 0, all donors received a DIB vaginal device (Syntex, Buenos Aires, Argentina) plus 2.5 mg estradiol-17β and 50 mg progesterone (P4; Laboratorio Rio de Janeiro, Argentina) IM. Superstimulatory treatments were initiated on Day 4, with a total dose of 320 mg (cows) or 200 mg (heifers) NIH-FSH-P1 of Folltropin-V (Bioniche Animal Health Inc., Belleville, Ontario, Canada) in twice daily IM injections over 4 days. All cows received prostaglandin-F treatment (150 µg d-cloprostenol; Bioprost; Biotay, Buenos Aires, Argentina) on Day 6 (AM and PM); DIB devices were removed with the last Folltropin-V treatment on Day 7 PM. Donors received 0.050 mg GnRH (Lecirelina; Gonasyn; Syntex, Buenos Aires, Argentina) or 25 mg Armour pLH (Lutropin-V; Bioniche) 12 h (Day 8 AM) or 24 h (Day 8 PM) later and were FTAI 12 and 24 h after GnRH or pLH. Ova/embryos were collected nonsurgically and classified according to IETS recommendations on Day 15 AM for donors receiving GnRH or pLH on Day 8 AM and on Day 15 PM for those receiving GnRH or pLH on Day 8 PM. Data were analyzed by two-way ANOVA. Although there was no effect of induction agent (GnRH vs. pLH; P > 0.2) and/or interaction between the induction agent and the time of treatment (P > 0.13) on ova/embryo production, treatment with GnRH or pLH on Day 8 AM (i.e. 12 h after DIB removal) resulted in a significantly lower (P < 0.05) number of fertilized ova and Grades 1 and 2 embryos than treatment on Day 8 PM (i.e. 24 h after DIB removal; Table 1). It was concluded that GnRH and pLH were equally efficacious for inducing ovulation in superstimulated donors that were fixed-time AI. Furthermore, a 24-h interval from progestin device removal to GnRH or pLH administration was preferable for Bos taurus beef donors. Table 1. Ova/embryo production in Angus donors treated with GnRH or pLH 12 or 24 h after removal of a progestin-releasing device and FTAI 12 and 24 h later

Various intravaginal progesterone-releasing devices are commercially available and each is impregnated with different amounts of progesterone. An experiment was designed to characterize plasma progesterone release profiles from Cue-Mate in comparison with other commercially available progesterone-releasing devices. Cycling, lactating Holstein cows (n = 60; BCS between 2.0 and 3.0 out of 5) from commercial dairy farms in Argentina were used in 2 replicates. All cows received 2 injections of D+ cloprostenol (PGF, Bioprost-D�; Biotay, Buenos Aires, Argentina) 14 days apart and were randomly assigned to one of four groups to receive intravaginal devices as follows: Cue-Mate (1.56 g progesterone; Bioniche Animal Health, A/Asia Pty, Armidale, Australia; n = 10), DIB (1 g progesterone; Syntex SA, Argentina; n = 10), CIDR with 1.9 g progesterone (Pfizer Salud Animal S.A., Buenos Aires, Argentina; n = 10), or CIDR with 1.38 g progesterone (Pfizer Animal Health, Groton, CT, USA; n = 10). All devices were inserted 24 hours after the second PGF and were left in place for 31 days. Blood samples were taken for progesterone analysis at 6-hour intervals for the first 24 hours, then daily for a week, and every 2 days for an additional 25 days. Plasma progesterone concentrations were measured with a modified human double-antibody RIA kit (DPC Coat-A-Count�; Diagnostic Products Corporation, Los Angeles, CA, USA). Cows were observed twice daily to confirm that vaginal inserts were still in place. Time-series hormone data were analyzed using the MIXED procedure for repeated measures (SAS Institute, Inc., Cary, NC, USA). Main effects of device and time, the highest mean concentration of progesterone, and the mean day on which this occurred were compared by a protected LSD test. There was no evidence of cow discomfort with any of the vaginal devices, and loss rates did not differ. There was an effect of group (P < 0.001) and day (P < 0.001) for progesterone concentrations, but there was no interaction (P = 0.89). Peak concentrations of progesterone (ng mL-1) did not differ among groups (Cue-Mate: 5.0 � 1.1; DIB: 3.9 � 0.6; 1.9-g CIDR: 4.6 � 0.6; 1.38-g CIDR: 3.7 � 0.4; P = 0.51). Mean (� SEM) concentrations of progesterone over the 31-day insertion period differed between 1.9-g CIDR (1.8 � 0.2 ng mL-1) and DIB (1.5 � 0.1 ng mL-1, P < 0.006), with Cue-Mate (1.5 � 0.2 ng mL-1) and 1.38-g CIDR (1.6 � 0.1 ng mL-1) intermediate and not different from either. When data were evaluated from insertion until Day 7, mean plasma progesterone concentrations did not differ among groups (Cue-Mate: 2.2 � 0.2 ng mL-1; DIB: 2.2 � 0.1 ng mL-1; 1.9-g CIDR: 2.6 � 0.2 ng mL-1; 1.38-g CIDR: 2.1 � 0.1 ng mL-1). However, progesterone concentrations from insertion until Day 21 were higher (P < 0.03) in cows with a 1.9-g CIDR (2.1 � 0.1 ng mL-1) than in those with a DIB (1.6 � 0.1 ng mL-1) or a 1.38-g CIDR (1.6 � 0.1 ng mL-1), with Cue-Mate (1.7 � 0.1 ng mL-1) intermediate and not different. Based on progesterone release patterns, it was concluded that Cue-Mate intravaginal devices can be used in protocols for synchronization of estrus and ovulation as with other progesterone-releasing devices.

High temperatures can be harmful to the competence of cumulus–oocyte complexes and to embryo development (Al-Katanani et al. 2002 J. Dairy Sci. 85, 390–396). The aim of this study was to evaluate the effect of maternal heat stress on in vitro embryo yield. Ten multiparous nonlactating Gir (Bos indicus) cows were kept in tie stalls for an adaptive period of 28 days [pre-heat-stress period (PRE-HS)/Days -28 to -1]. Cows were subjected to 2 OPU (ovum pickup) sessions (Days -14 and -7). In the heat-stress period (HS; Days 0 to 28), cows were divided into control (C: n = 5) and heat-stressed (HS: n = 5) groups. During this period, OPU sessions were performed once a week from Days 0 to 28. The C group remained in a thermo-neutral environment, and the HS group was kept in a climatic chamber with controlled temperature and humidity (38�C and 80% during the day and 30�C and 80% during the night). In the post-heat-stress period (POST-HS; Days 28 to 147), all cows returned to thermo-neutral conditions. Then 17 OPU sessions were performed once a week from Days 35 to 147. In all periods, blood samples were collected weekly for progesterone (P4) analysis, and ovarian follicles were counted, measured, and aspirated. The COCs were evaluated and selected for the IVF procedure. Data were analyzed by ANOVA (PROC MIXED of SAS) and a chi-squared test. The luteal phase was defined as the period between 2 samples with P4 below 1.0 ng mL-1. A handling accident caused the exclusion of an HS cow after the sixth session. The C and HS groups were subjected to 125 and 107 OPU sessions, respectively. Means � SEM for the C vs. HS groups, in the PRE-HS, HS, and POST-HS periods, respectively, were visualized follicles: 25.5 � 2.5 vs. 28.5 � 2.8, 24.2 � 1.1 vs. 24.0 � 1.9, and 15.3 � 0.6 vs. 15.8 � 0.8; largest follicle diameter: 12.1 � 1.5 vs. 11.1 � 1.7, 13.3 � 0.8 vs. 13.0 � 0.6, and 11.4 � 0.4b vs. 14.0 � 0.4a; P < 0.05; 2nd largest follicle diameter: 6.2 � 1.3 vs. 6.0 � 1.2, 5.9 � 0.6 vs. 7.1 � 0.8, and 6.3 � 0.3b vs. 8.7 � 0.5a; recovered COCs: 11.2 � 2.8 vs. 14.3 � 2.5, 9.6 � 1.0 vs. 11.0 � 1.3, and 8.6 � 0.7 vs. 7.9 � 0.6; COCs selected for IVF: 69/112 (61.6%)b vs. 108/143 (75.5%)a, 164/241 (68%) vs. 172/265 (64.9%), and 426/712 (59.8%) vs. 305/535 (75.0%); cleavage: 44/59 (74.5%) vs. 87/105 (82.9%), 72/101 (71.3%) vs. 74/121 (61.2%), and 226/317 (71.3%) vs. 159/230 (69.1%); embryos per cow/OPU: 2.1 � 1.1y vs. 4.1 � 1.0x, 0.4 � 0.3 vs. 0.5 � 0.3, and 0.9 � 0.2x vs. 0.4 � 0.1y; P < 0.1; and blastocyst yield: 16/59 (27.1%) vs. 33/105 (31.5%), 11/31 (35.5%) vs. 13/52 (25.0%), and 76/279 (27.2%)a vs. 25/188 (13.3%)b. In conclusion, maternal heat stress increased the percentage of short estrous cycles, decreased the P4 concentrations, and decreased the number of embryos produced by Bos indicus cows, mainly from 28 to 147 days post-heat-stress, showing long-term deleterious effects on blastocyst development.

Three experiments were designed to evaluate the effect of the duration of treatment with progesterone (P4) vaginal devices (1 g of P4, Triu-B; Biog�nesis, Buenos Aires, Argentina) on pregnancy rates in crossbred zebu cattle artificially inseminated at a fixed time (FTAI). In Exp. 1, 371 non-lactating multiparous crossbred zebu cows with a body condition score (BCS) between 2.0 and 3.0 out of 5 were randomly allocated to one of nine treatment groups in a 3 � 3 factorial design. On Day 0, all cows received 2 mg of estradiol benzoate (EB, Bioestrogen; Biog�nesis) i.m. and were divided into groups to receive a new Triu-B device, a once-previously-used Triu-B (2nd-use), or a Triu-B that was previously used twice (3rd-use) and in which three silicone rings (2 cm in length) impregnated with 100 mg P4 each were threaded through the wings of the device. Triu-B devices are currently approved for re-use up to three times. Cows were further subdivided to have their Triu-B devices removed on Days 7, 8, or 9. PGF (150 �g D(+)cloprostenol, Enzaprost-DC; Biog�nesis) was administered i.m. at the time of Triu-B removal, followed by 1 mg EB 24 h later; all cows were FTAI 52 to 56 h after Triu-B removal. In Exp. 2, 417 crossbred zebu heifers, 20 to 26 months of age and BCS between 2.5 and 3.5, were treated similarly to the cows in Exp. 1, except that only new or 3rd-use Triu-B were used. In Exp. 3, 836 crossbred zebu heifers, similar to those used in Exp. 2, were also treated similarly to the cows in Exp. 1, but in this case, Triu-B devices were inserted for 8, 9, or 10 days. Pregnancy rates were determined by ultrasonography 50 to 60 days after FTAI and were analyzed by logistic regression. In Exp. 1, there were no significant effects of duration of treatment [7 days: 69/137 (50.4%), 8 days: 68/139 (48.9%), or 9 days: 41/95 (43.2%); P = 0.28], or type of Triu-B used (P = 0.91; new Triu-B: 47.7%, 2nd-use: 48.9%, or 3rd-use: 47.2%). In Exp. 2, there were no significant effects of duration of treatment [7 days: 76/139 (54.7%), 8 days: 63/143 (44.1%), or 9 days: 73/135 (54.1%); P = 0.98] or type of Triu-B used (new Triu-B: 48.6% and 3rd-use Triu-B: 53.2%; P = 0.33). In Exp. 3, there was an interaction between duration of treatment and pregnancy rates (P < 0.05). Lower pregnancy rates were obtained in 9- and 10-day treatments than in 8-day treatments, but only when previously used Triu-B devices were used. Pregnancy rates were: new Triu-B for 8 days: 42/89 (47.2%), 9 days: 37/90 (41.1%), and 10 days: 41/91 (45.1%); 2nd-use Triu-B for 8 days: 51/103 (49.5%), 9 days: 39/104 (37.5%), and 10 days: 39/109 (35.8%); 3rd-use Triu-B for 8 days: 52/90 (57.4%), 9 days: 31/90 (34.4%), and 10 days: 25/70 (35.7%); P < 0.01. Results demonstrate that treatment of crossbred zebu cattle with P4 devices for 7 to 10 days results in comparable pregnancy rates when new Triu-B devices are used. However, pregnancy rates decreased when previously used Triu-B devices were inserted for more than 8 days.

An experiment was designed to determine plasma progesterone (P4) concentrations in ovariectomized cows treated with either CIDR-B devices (Pfizer Animal Health, Hamilton, New Zealand) impregnated with 1.9 g of P4 or DIB devices (Syntex, Argentina) impregnated with 0.5 or 1.0 g of P4. Fifteen ovariectomized beef cows, weighing 350 to 450 kg and with a body condition score (BCS) 2.5 to 3.0 out of 5 were used. All cows were ovariectomized 30 days before the beginning of the trial and had plasma P4 concentrations 0.13) or a day by treatment interaction (P > 0.16; Table 1). The area under the curve did not differ (P = 0.95) among groups (DIB 0.5: 2236.4 ng, DIB 1.0: 2164.6 ng, and CIDR-B: 2266.8 ng). In all treatments, plasma P4 profiles were characterized by a sudden increase in P4 concentrations within 12 h of device insertion and a gradual decrease during the following several days. It was concluded that the use of CIDR-B devices impregnated with 1.9 g of P4 or DIB devices impregnated with 1.0 or 0.5 g of P4 did not result in different plasma P4 concentrations in ovariectomized cows. Further studies are needed to confirm these results in lactating and dry intact (not ovariectomized) cattle. Table 1. Mean plasma P4 concentrations (ng/mL ± SEM) in ovariectomized cows treated with intravaginal devices containing 0.5 g (DIB 0.5), 1.0 g (DIB 1.0), or 1.9 g (CIDR-B) of P4*

Three experiments were conducted to compare pregnancy rates in cows and heifers treated with intravaginal progesterone (P4)-releasing devices impregnated with either 0.5 or 1.0 g of P4 and inseminated at a fixed time (FTAI). In Experiment 1, 102 postpartum beef cows (Hereford and Angus crossbred with Tuli, 60 to 90 d postpartum) with a body condition score (BCS) of 2.5 to 3.0 (1 to 5 scale) were used. On Day 0, all cows received 2 mg of estradiol benzoate (EB; Syntex, Argentina) i.m. and were randomly assigned to receive an intravaginal progesterone-releasing device with 0.5 g of P4 (DIB 0.5 g; Syntex) or 1.0 g of P4 (DIB 1.0 g; Syntex). On Day 7, DIB devices were removed and all cows received 150 µg D(+)cloprostenol (Ciclase; Syntex) i.m. On Day 8, all cows received 1 mg EB and were FTAI 52 to 56 h after DIB removal. In Experiment 2, 93 Holstein cows that were 90 to 110 days in milk (DIM), producing on average 5000 kg of milk per lactation and with a BCS of 2.5 to 3.0 received the same treatments as those in Experiment 1. In Experiment 3, 100 Holstein heifers, 20 to 22 months old, with a BCS of 2.5 to 3.0, also received the same treatments as those in Experiment 1. Pregnancy rates in all experiments were determined by ultrasonography 35 days after FTAI and compared by chi-square test. No differences in pregnancy rates were detected between lactating beef cows, dairy cows, or dairy heifers treated with DIB devices impregnated with 0.5 or 1.0 g of P4 (Table 1). It was concluded that vaginal P4 releasing devices impregnated with 0.5 g or 1.0 g of P4 results in comparable pregnancy rates in FTAI protocols in beef and dairy cattle. Table 1. Pregnancy rates in postpartum beef cows, Holstein cows, and Holstein heifers treated with DIB devices impregnated with 0.5 or 1.0 g of P4 and subjected to FTAI

Two experiments were designed to investigate the use of eCG in a fixed-time embryo transfer (FTET) protocol for Chinese Yellow crossbred recipients receiving in vitro-produced (IVP) bovine embryos. In Experiment 1, cows were observed for spontaneous estrus (Group 1) or following 500 �g cloprostenol (PGF; Schering-Plough Animal Health, Montreal, Quebec, Canada; Group 2) with nonsurgical embryo transfer 7.5 or 8.0 days later. Cows in Groups 3, 4, and 5 were treated with a CIDR insert (Bioniche Animal Health, Beijing, China), 2 mg estradiol benzoate (EB), and 50 mg progesterone (Sigma-Aldrich, Canada) i.m. on Day 0, PGF on Day 5, CIDR removal on Day 7, and 1 mg EB i.m. 24 h later with nonsurgical transfer of Holstein IVP embryos 8.5 days later, without estrus detection. In addition, cows in Group 3 received no further treatment, whereas cows in Group 4 received 400 IU eCG i.m. (Pregnecol; Bioniche) on Day 5, and cows in Group 5 received eCG on Day 7. Holstein embryos were produced utilizing slaughterhouse ovaries and standard IVF procedures. Expanded blastocysts of quality grade 1 (IETS) were cryopreserved in 10% ethylene glycol and 20% fetal bovine serum using standard procedures. Straws were thawed in a 30�C water bath, and embryos were expelled directly into holding medium and evaluated prior to nonsurgical transfer (NT) ipsilateral to the corpus luteum (CL). Pregnancy was diagnosed ultrasonically 30 d later. Although pregnancy rates, based on numbers of recipients synchronized, numerically favored the administration of eCG on Day 5, there were no significant differences (P = 0.40) among groups (Group 1: 15.4%, n = 52; Group 2: 20.0%, n = 50; Group 3: 19.2%, n = 99; Group 4: 28.1%, n = 96; Group 5: 21.3%, n = 75). In Experiment 2, Chinese Yellow crossbred cattle were synchronized with a CIDR insert plus estradiol and progesterone on Day 0, PGF on Day 5, CIDR removal on Day 7 and estradiol on Day 8, and were randomly assigned to received no further treatment (Group 1; n = 400) or an injection of 400 IU eCG on Day 5 (Group 2; n = 391). Recipients with a detectable CL received a frozen-thawed IVP Holstein embryo by NT 8.5 days after the second injection of estradiol without estrus detection, as in the first experiment. Pregnancy diagnosis was done ultrasonically 30 days later. Although CL size, cow age, and embryo quality, prior to transfer, were recorded, no effects on pregnancy rates were demonstrated (P = 0.30). Pregnancy rates, based on recipients receiving embryos, did not differ (P = 0.5) between groups (Group 1: 21.4%, n = 154; Group 2: 24.5%, n = 290). Overall pregnancy rates (based on the total number of recipients synchronized) were significantly higher (P < 0.001) in Group 2 (eCG; 18.2%) than in Group 1 (no eCG; 8.3%), because of the significantly higher (P < 0.03) percentage of recipients used following treatment with eCG (74.2% vs. 38.5%). Results indicate that the administration of eCG on Day 5 of an 8-d synchronization protocol for FTET of frozen-thawed bovine IVP embryos will improve pregnancy rates in beef recipients, especially those of marginal quality.

Two experiments were designed to compare the effects of eCG treatment and temporary weaning on ovulation and pregnancy rates in postpartum cows. In Experiment 1, 39 lactating multiparous crossbred Bos indicus cows, 60 to 80 d postpartum with a body condition score (BCS) between 2.0 to 2.5 out of 5 were randomly allocated to one of four treatment groups in a 2 � 2 factorial design. On Day 0, all cows received a progesterone (P4) intravaginal device (DIB; Syntex, Argentina) and 2 mg estradiol benzoate (EB) i.m. (Syntex). On Day 8, DIB devices were removed and all cows received 150 �g D(+)cloprostenol i.m. (Ciclase; Syntex) and were randomly divided to receive 400 IU eCG i.m. (Novormon 5000; Syntex) at the same time or no treatment. In addition, half of the cows in each group had their calves weaned temporarily (TW) for 56 h at the time of DIB removal; the other half remained with their calves. All cows received 1 mg EB i.m. on Day 9 and were examined every 8 h by ultrasonography from the time of DIB removal until ovulation. Data were analyzed by ANOVA and chi-square test. The diameter of the dominant follicle on Day 8 (eCG: 7.8 � 0.5 mm vs. no eCG: 8.2 � 0.4 mm, and TW: 7.6 � 0.4 mm vs. no TW 8.4 � 0.5 mm), the proportion of cows that ovulated (eCG: 12/20, 60% vs. no eCG: 9/19, 47%; and TW: 13/20, 65% vs. no TW: 8/19, 42%, and the interval to ovulation (eCG: 72.0 � 1.4 h vs. no eCG: 75.6 � 2.0 h, and TW: 73.8 � 1.6 h vs. no TW: 73.0 � 1.8 h) did not differ among groups (P > 0.05). Although there was no effect of eCG treatment or eCG by TW interaction (P > 0.3) on the size of the preovulatory follicle (eCG: 11.1 � 0.4 mm vs. no eCG: 10.1 � 0.6 mm), it was smaller in cows TW (9.9 � 04 mm), compared to those not TW (11.8 � 0.3 mm; P < 0.05). Nevertheless, the growth rate of the ovulatory follicle was greater (P < 0.02) in cows treated with eCG (1.1 � 0.1 mm/day) than in those not treated with eCG (0.6 � 0.1 mm/day). In Experiment 2 769 lactating multiparous crossbred Bos indicus cows received the same treatments as those in Experiment 1 and were fixed-time inseminated (FTAI) 52 to 56 h after DIB removal. Cows were examined by ultrasonography on Day 0 (22.5% with corpora lutea (CL), 30.0% with follicles >8 mm, and 47.5% with follicles 0.7) by TW (TW: 141/397, 37.2% vs. no TW: 141/390, 36.1%) or the TW by eCG interaction (P > 0.7). However, pregnancy rates were higher (P < 0.05) in cows treated with eCG (154/377, 40.8%) than in those not treated with eCG (128/392, 32.6%). The use of eCG but not TW improved pregnancy rates following FTAI in postpartum Bos indicus cows. Results also suggest that the eCG-related increase in pregnancy rates may be due to the final growth rate of the ovulatory follicle.

An experiment was designed to evaluate the effect of the time of removal of an intravaginal progesterone (P4) releasing device and GnRH treatment on the interval to and the distribution of ovulations in beef cows superstimulated with Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada). Red Angus donors (22 cows and 12 heifers) with a body condition score between 2.5 to 3.5 out of 5 were blocked by parity and randomly assigned to one of four treatment groups in a 2 × 2 factorial design. On Day 0, all donors received an i.m. DIB vaginal device (Syntex, Argentina) plus 2.5 mg estradiol-17β and 50 mg P4 (Laboritorio Rio de Janeiro, Argentina). Superstimulatory treatments were initiated on Day 4, with a total dose of 320 mg (cows) or 200 mg (heifers) NIH-FSH-P1 (Folltropin-V; Bioniche Animal Health, Inc., Belleville, Ontario, Canada) in twice daily i.m. injections over 4 days. All cows received PGF treatment (150 μD(+)cloprostenol: Ciclase; Syntex) in the morning and afternoon of Day 6 and were randomly divided to have DIB devices removed at the time of the second PGF treatment (Day 6.5) or 12 h later (Day 7). On Day 8, donors were further subdivided to receive GnRH (0.050 mg Lecirelina, Gonasyn; Syntex) or no further treatment, and all cows were fixed-time AI either 60 and 72 h after the first PGF injection (0 h). All cows and heifers were examined by ultrasonography every 8 h for a period of 120 h commencing at the time of DIB removal, to determine the time and distribution of ovulations. For each cow, mean time of ovulation was used for the statistical analysis. The distributions of ovulations were compared among groups by Levene's test, and then by ANOVA (equal variances) or Friedman two-way non-parametric ANOVA (unequal variances). No significant differences were detected between cows and heifers (P > 0.13) for the parameters evaluated, and data were combined. There was no effect (P > 0.22) of treatment on the number of ovulations (GnRH: 12.6 ± 1.6 vs. no GnRH: 15.1 ± 1.1 and Day 6.5: 13.7 ± 1.3 vs. Day 7: 14.3 ± 1.5), or numbers of transferable embryos (GnRH: 5.1 ± 1.1 vs. no GnRH: 3.1 ± 0.8 and Day 6.5: 3.5 ± 0.8 vs. Day 7: 4.81 ± 0.1). However, there was a significant effect of GnRH treatment and a GnRH-by-time-of-DIB-removal interaction on the distribution of ovulations (P < 0.05; Table 1). The administration of GnRH in a superstimulatory protocol using P4 releasing devices results in earlier and more synchronous ovulations than when GnRH is not used. Table 1. Distribution of ovulations (hours after the first PGF) in superstimulated Red Angus donors treated with progesterone-releasing devices (DIB) for 6.5 or 7 days with and without GnRH treatment on Day 8

Two experiments were designed to evaluate the effect of different treatments for the synchronization of ovulation on pregnancy rates following fixed-time AI (FTAI) of lactating dairy cows. In Experiment 1, 394 Holstein cows that were 61.7 � 13.6 days postpartum (range 35 to 94 days), with a milk yield of 30.7 � 6.8 liters per day (range 12.0 to 52.4 liters) and a body condition score (BCS) between 2.5 to 3.5 out of 5, were used. Cows were blocked by days postpartum and milk yield and randomly assigned to one of four treatment groups. Cows in P4+EB treatment groups received an intravaginal DIB device (1 g P4; Syntex, Argentina) and 2 mg of estradiol benzoate (EB; Syntex) i.m. on Day 0. On Day 8, DIB devices were removed, and cows received PGF (150 �g D+cloprostenol: Ciclase; Syntex), and were subdivided to receive 400 IU eCG (Novormon 5000, Syntex) i.m. or no further treatment at that time. On Day 9, all cows received 1 mg of EB and were FTAI 60 h after DIB removal. Cows in P4-Synch groups received a DIB device and 50 �g of GnRH (Lecirelina, Gonasyn; Syntex) i.m. on Day 0. On Day 7, DIB devices were removed; cows received PGF and were divided to receive 400 IU eCG i.m. or no further treatment. On Day 9, all cows received a second GnRH treatment and were FTAI 60 h after DIB removal. Blood samples were taken on Days -10 and 0 to determine plasma P4 concentrations; 93% of the cows had >1 ng/mL P4 in at least one sample. In Experiment 2, 200 lactating cows from the same farm were treated with the P4+EB+eCG and P4-Synch without eCG treatments with either a DIB or a CIDR-B (1.9 g P4; Pfizer Animal Health, Groton, CT, USA) in a 2 � 2 factorial design. Cows were examined by rectal palpation 50 days after FTAI to determine pregnancy status, and data were analyzed by chi-square Mantel-Haenszel test. There was an EB/GnRH by eCG interaction (P < 0.05) which was attributed to a higher pregnancy rate in the P4+EB+eCG group (44/98; 44.9%) than in the P4+EB without eCG group (30/100; 30.0%) and P4-Synch+eCG group (30/98; 30.6%); the P4-Synch without eCG group had an intermediate pregnancy rate that did not differ from the other treatment groups (37/98; 38.8%). In Experiment 2, no significant differences (P = 0.40) in pregnancy rates were detected between cows treated with DIB (51/100; 51.0%) or CIDR-B (42/100; 42.0%), and there was no difference (P = 0.18) between P4+EB+eCG (52/100; 52.0%) and P4-Synch without eCG (41/100; 41.0%) treated groups. Results suggest that the addition of eCG will improve pregnancy rates following FTAI in lactating dairy cows treated with EB but not in those treated with GnRH at the time of insertion and after removal of a P4 releasing device. Although treatment with P4+EB+eCG resulted in numerically higher pregnancy rates, results were not different from those obtained in the GnRH-based treatment, without the addition of eCG.

Although treatments with progesterone (P4) releasing devices, estradiol benzoate (EB) and eCG have been shown to result in acceptable pregnancy rates after embryo transfer, the treatment requires that the cows be run through the chute at least four times for treatments. An experiment was designed to compare pregnancy rates in cows treated with P4 releasing devices plus EB and eCG, and induced to ovulate with EB given at device removal or 24 h later. Non-lactating Bos taurus × Bos indicus crossbred beef cows (n = 165), with a body condition score between 2.5 to 3.5 (1 to 5 scale), were treated with a P4-device (DIB, Syntex, Argentina) and 2 mg EB i.m. (Syntex), on Day 0 and 400 IU of eCG i.m. (Novormon, Syntex) plus 150 μg D(+)cloprostenol i.m. (Ciclase, Syntex) on Day 5. On Day 8, DIB devices were removed and cows were randomly divided into two groups to receive either 1 mg EB i.m. at the time of DIB removal (EB0) or 24 h later (Day 9; EB24). Recipients were observed for signs of estrus for 48 h after DIB removal. On Day 16, all recipients observed in estrus and with >1 CL or a single CL with an area >256 mm2 were selected to receive fresh embryos on Day 16 (EB0) or Day 17 (EB24). Furthermore, 20 recipients not observed in estrus but with a CL >256 mm2 were randomly selected and transferred. The embryos used were 28 Grade 1, 40 Grade 2, and 24 Grade 3. Ovarian ultrasonography was performed on Day 0, to determine ovarian status (only cows with a CL or a follicle >10 mm and uterine tone were used), on Day 16 to measure CL area, and 60 days after embryo transfer to determine pregnancy status. Quantitative data were analyzed by Student's t-test and qualitative data were analyzed by logistic regression. There were no differences between groups in the mean (± SEM) CL area on Day 16 (EB0: 300.3 ± 12.0 mm2 and EB24: 324.9 ± 11.7 mm2; P = 0.14), the proportion of recipients in estrus (EB0: 38/82, 46.3%, and EB24: 34/83, 41.0%; P = 0.49), the number of recipients with >1 CL or a CL >256 mm2 (EB0: 68/82, 82.9%, and EB24: 72/83, 86.7%; P = 0.49) and the number of recipients pregnant/transferred (EB0: 31/49, 63.4%, and 20/43, 46.5%; P = 0.23). Furthermore, there were no significant effects of embryo quality (P = 0.31) or technician (P = 0.12) on pregnancy rates. The mean interval from DIB removal to estrus was shorter (P = 0.001) for recipients in the EB0 group (22.7 ± 1.0 h) than for those in the EB24 group (37.4 ± 1.2 h). Nevertheless, pregnancy rates did not differ (P = 0.14) between recipients seen in estrus (EB0: 21/38, 55.3%, and EB 24: 16/34, 47.1%) and those not seen in estrus but with a CL >256 mm2 on Day 16 (EB0: 10/11, 90.9%, and EB24 4/9, 44.4%). It was concluded that the two treatments evaluated are equally efficacious for synchronizing Bos taurus × Bos indicus recipients. Furthermore, the use of EB at device removal could reduce the number of trips through the chute without affecting pregnancy rates. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.

Although treatments with progesterone (P4)-releasing devices and estradiol benzoate (EB) have been extensively used in fixed-time AI (FTAI) programs in beef cattle, pregnancy rates in postpartum Bos indicus cows kept on pasture often have been lower than expected because of poor body condition score (BCS) and a high incidence of anestrus. Temporary weaning and eCG treatment have been shown to increase pregnancy rates in suckled beef cows. Therefore, this experiment was designed to compare the effect of eCG treatment and temporary weaning on pregnancy rates in postpartum cows in fair to poor BCS. Lactating multiparous crossbred Bos indicus cows (n = 393), 60 to 90 d postpartum with a BCS 2.0 (1 to 5 scale) were randomly allocated to 1 of 4 treatment groups, in a 2 × 2 factorial design. At the beginning of the experiment (Day 0), all cows received a P4 intravaginal device (DIB, Syntex, Argentina) and 2 mg EB i.m. (Syntex). On Day 8, DIB devices were removed and all cows received 150 μg D (+) cloprostenol i.m. (Ciclase, Syntex) and were randomly divided to receive 400 IU eCG (Novormon 5000, Syntex) or no treatment at the same time. Furthermore, half of the cows in each treatment group have their calves weaned temporarily, from the time of DIB removal until the end of the FTAI, whereas the other half remained with their calves all the time. Finally, all cows received 1 mg EB i.m. on Day 9 and were FTAI 52 to 56 h after DIB removal. Cows were examined by ultrasonography 42 d after FTAI to determine pregnancy status. Data were analyzed by logistic regression and the effects of treatment and semen used were considered in the model. Ovarian activity was estimated by rectal palpation on Day 0: there were 72/393 (18.2%) cows with a CL, 140/393 (35.6%) cows with palpable medium size follicles, and 181/393 (46.1%) cows with ovaries containing no detectable structures. There was no effect of ovarian status at the time of treatment (P = 0.91) or semen (P = 0.91) on pregnancy rates. Treatment with eCG tended (P = 0.08) to increase pregnancy rates (94/192, 48.9% and 79/201, 39.3% for cows treated or not treated with eCG, respectively). However, there was no effect of weaning or eCG by weaning interaction (P = 0.98) on pregnancy rates (86/191, 45.1% and 87/202, 43.1% for cows that have their calves weaned or not weaned for 56 h, respectively). Results confirm those of previous studies that demonstrated that the use of eCG in a P4/EB/FTAI program improved pregnancy rates in postpartum Bos indicus crossbred cows that were in fair-to-poor BCS. However, no improvement in pregnancy rates was observed after temporary weaning in the present study. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.

Previous studies have shown that the use of eCG given at the time of removal of a progesterone (P4) releasing device improved pregnancy rates to fixed-time AI (FTAI) in postpartum Bos indicus cows (Bo GA et al. 2004 Reprod. Fertil. Dev. 16, 127). Suckling restriction with the placement of nose tags in calves has also been shown to induce cyclicity and increase pregnancy rates in postpartum Bos indicus cows (Stahringer RC 2003 Taurus 18, 21–23). An experiment was designed to compare the effect of eCG treatment and restricted suckling on pregnancy rates in postpartum cows in fair to poor body condition score (BCS). A secondary objective was to evaluate the effect of restricted suckling on weaning weights. Lactating primiparous crossbred Bos indicus cows (n = 399), 60 to 90 days postpartum with a BCS 2.0 (1 to 5 scale), were randomly allocated to 1 of 4 treatment groups, in a 2 × 2 factorial design. On Day 0, all cows received a P4 intravaginal device (DIB, Syntex, Argentina) and 2 mg estradiol benzoate (EB) i.m. (Syntex). On Day 8, DIB devices were removed and all cows received 150 μg D(+)cloprostenol i.m. (Ciclase, Syntex) and were randomly divided to receive 400 IU eCG (Novormon, Syntex) or no treatment at the same time. On Day 9, all cows received 1 mg EB i.m. and were FTAI 52 to 56 h after DIB removal. Nose tags were placed in half of the calves from Day 0 to the time of FTAI, whereas the other half of the calves remained untreated. All cows were examined by ultrasonography 30 d after FTAI to determine pregnancy status. Pregnancy data were analyzed by logistic regression, and birth and weaning weights were analyzed by Student's t-test. Ovarian activity was estimated by rectal palpation on Day 0: there were 57/399 (14.3%) cows with a CL, 203/399 (50.8%) cows with palpable medium size follicles, and 139/399 (34.8%) cows with ovaries containing no detectable structures. There was no effect of ovarian status at the time of treatment (P = 0.52) or semen (P = 0.89) on pregnancy rates. Suckling restriction increased (P = 0.03) pregnancy rates (91/195, 46.6% and 81/204, 39.7% for cows not suckled or suckled during the treatment, respectively). However, there was no effect of eCG or eCG by weaning interaction (P = 0.82) on pregnancy rates (82/192, 42.7% and 90/207, 43.4% for cows treated or not treated with eCG, respectively). Although mean (±SEM) birth weights were not different (29.4 ± 0.4 vs. 29.1 ± 0.4; P = 0.6), weaning weights were significantly affected (P = 0.001) by the suckling restriction (190.1 ± 1.9 vs. 200.4 ± 2.1 for calves treated or not treated with the nose tags, respectively). Although restricting suckling, by placement of a nose tag from Day 0 to FTAI, may increase pregnancy rates in primiparous Bos indicus cows in fair to poor BCS, the adverse effect of this treatment on weaning weights of the calves has to be considered. Furthermore, eCG did not increase pregnancy rates in the present study, as it has been previously reported. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.

Although several studies have investigated the relationship between circulating progesterone and pregnancy rates in cattle, the beneficial effect of treatments that increase progesterone concentrations, by insertion of a progesterone (P4) releasing device or induction of an accessory CL with hCG, GnRH, or LH treatment, has resulted in inconsistent effects on pregnancy rates in embryo recipients. An experiment was designed to evaluate the effect of hCG or GnRH treatment, given at the time of embryo transfer without estrus detection, on pregnancy rates in recipients treated with intrauterine P4-releasing devices, estradiol benzoate (EB), and eCG. The experiment was performed in two replicates; non-lactating Bos taurus × Bos indicus crossbred beef cows with a body condition score between 2.5 to 3.5 (1-to-5 scale) were used (replicate 1, n = 180; replicate 2, n = 140). All cows received 1 g of P4 via a P4-releasing device (DIB, Syntex, Argentina) and 2 mg EB i.m. (Syntex) on Day 0, and 400 IU of eCG i.m. (Novormon 5000, Syntex) plus 150 μg d(+)cloprostenol i.m. (Ciclase, Syntex) on Day 5. DIBs were removed on Day 8 and all cows received 1 mg EB i.m. on Day 9. Recipients were not observed for signs of estrus, and those >1 CL, or a single CL with an area >256 mm2, received 195 Grade 1 and 46 Grade 2 frozen/thawed “direct transfer” embryos on Day 17. At the time of embryo transfer, recipients were randomly allocated to 1 of 3 treatment groups to receive 1500 IU hCG (Ovusyn, Syntex), 50 μg Lecirelina (GnRH, Gonasyn, Syntex), or no treatment (control) at that time. Ovarian ultrasonography was performed on Day 0 to determine ovarian status (only cows with a CL or a follicle >10 mm and uterine tone were used), on Day 17 to measure CL area, and 40 days after embryo transfer to determine pregnancy status. Data were analyzed by logistic regression and the effects of replication, technician, treatment, and embryo quality were considered in the model. From the 320 recipients treated with a DIB plus EB and eCG, 241 (75.3%) were selected to receive an embryo. Nine (3.7%) and 1 (0.4%) of the selected recipients had 2 and 3 CL, respectively. Pregnancy rates did not differ between replicates (replicate 1: 80/140, 57.1%; and replicate 2: 57/101, 56.4%; P = 0.84), technicians (technician 1: 65/118, 55.1%; and technician 2: 72/123, 58.5%; P = 0.64), or treatments (hCG: 43/80, 53.8%; GnRH: 45/83, 54.2%; and control: 49/78, 62.8% P = 0.99). However, pregnancy rates were higher (P = 0.001) in recipients receiving Grade 1 embryos (121/195, 62.1%) than in those receiving Grade 2 embryos (16/46, 34.8%). GnRH or hCG treatment at the time of embryo transfer did not increase pregnancy rates in recipients synchronized with P4 releasing devices, EB, and eCG. Research was supported by Syntex S.A., Estancia El Mangrullo S.A., and Agencia Cordoba Ciencia S.E.

Estradiol and progesterone have been used to synchronize follicular wave emergence and ovulation in a two-dose prostaglandin (PGF)-based synchronization program (Martínez et al. 2004 Theriogenology 62, 363–372). However, it was observed that some heifers displayed estrus prior to the second PGF, suggesting that premature luteolysis may have occurred. An experiment was designed to determine the effects of dose of estradiol benzoate (EB) and/or progesterone (P) on follicular and luteal dynamics in a two dose PGF-based protocol in beef heifers. In two replicates, beef heifers (n = 28; Simmental, Hereford, and Charolais crosses, 350 to 450 kg) received 500 μg cloprostenol (Schering-Plough Animal Health, Pointe-Claire, PQ, Canada) on Day −7. On Day 0, heifers were randomly allocated to nine treatment groups to receive 0, 1, or 2 mg of EB and 0, 50, or 100 mg of P i.m. in canola oil in a 3 × 3 factorial design. A second PGF treatment was administered on Day 14. Ultrasonography was done once daily from Days −5 to 9, and every 12 h thereafter until ovulation. Blood samples were collected at 12-hour intervals from Day 0 to 5 for estradiol and FSH concentrations, and every 24 h for progesterone. The effects of EB and P and their interaction on corpus luteum (CL), follicles, and hormone profiles were analyzed by analysis of variance, and means were compared by LSD or Tukey's test. All variables were normally distributed (Wilk-Shapiro test and rankit plots). The day of follicular wave at the time of treatment tended to vary among groups (P = 0.08) and the diameter of the dominant follicle also differed (P < 0.05). The interval from treatment to wave emergence was shorter (P < 0.05) in heifers that received 2 mg EB (4.6 ± 0.3 d) than in those that did not receive EB (5.9 ± 0.6 d), while the 1 mg EB group (5.1 ± 0.6 d) was intermediate. The interval to wave emergence in the 2 mg EB group was the least variable (P < 0.05). There was no effect of EB (P = 0.72) on the diameter of the CL at the time of the second PGF, but there was an effect of P treatment (P = 0.01). The variability of the interval from the second PGF to ovulation may have been influenced by treatment group but was statistically not significant (P < 0.1). There was an effect of time (P < 0.01) on plasma progesterone concentrations and a P × time interaction (P = 0.06) can also be assumed. Estradiol concentrations were affected by EB dose, time, and EB × time interaction (all P < 0.01). FSH concentrations were modified by time (P < 0.01). In summary, treatment with EB 7 days after a single injection of PGF affected follicular development, while P treatment at that time appeared to influence CL function.

It has been shown recently that treatments with progesterone (P4)-releasing devices combined with estradiol benzoate (EB) plus P4 on Day 0, eCG and PGF on Day 5 and a second application of EB one day after device removal (Day 9) can be used successfully to transfer bovine embryos at a self-appointed time, without the necessity of estrus detection. Although the treatment solved one of the major problems in recipient management, estrus detection, it requires handling the recipients at least five times for treatments and embryo transfer. An experiment was designed to evaluate whether reducing one day of handling, by the administration of eCG and PGF at the time of removal of the P4 device (Day 8), results in comparable pregnancy rates than giving eCG on Day 5. A secondary objective was to determine the effect of injectable P4 at the time of device insertion plus EB treatment. Crossbred Bos taurus × Bos indicus beef heifers (n = 301) were randomly assigned to 4 treatment groups in a 2 by 2 factorial design. All Heifers received a P4 device (DIB, Syntex, Argentina) plus 2 mg EB i.m. (Syntex) at unknown stages of the estrous cycle (Day 0), with or without 50 mg of P4 given i.m. at the same time. Heifers were further subdivided to receive PGF (0.150 mg d-cloprostenol, Prolise, Tecnopec, Sao Paulo, Brazil) and 400 IU of eCG (Novormon, Syntex) i.m. on Days 5 or 8. In all heifers, DIB devices were removed on Day 8 and 1 mg EB was administered i.m. on Day 9. Day 10 was arbitrarily considered as the day of estrus. On Day 17, heifers were bled for plasma P4 concentrations and examined by ultrasonography to determine the number of CL and their diameter. Heifers that had >1 CL or a single CL with diameter ≥18 mm received an in vitro-produced (IVP) embryo by nonsurgical transfer performed by the same veterinarian. Pregnancy rates were determined by ultrasonography 30 days later. The effects of Day of eCG administration (Day 5 or Day 8), P4 of treatment (E2 or E2 + P4) and the day-by-P4 treatment interaction on the numbers of CL and plasma P4 were analyzed by ANOVA, and the proportion of recipients selected and pregnant were analyzed using non-parametric tests (NPAR1WAY, SAS). There was no significant effect of P4 treatment or the P4-by-day of eCG interaction in any of the parameters evaluated. However, there was a significant effect of day of eCG administration on plasma P4 concentrations (Day 5 = 2.4 ± 0.3 v. Day 8 = 1.7 ± 0.2; P = 0.03) and the number of CL (Day 5 = 1.4 ± 0.1 v. Day 8 = 1.1 ± 0.0; P = 0.02) on Day 17. Furthermore, the proportion of recipients pregnant/treated tended (P = 0.1) to be higher in heifers in the Day 5 Group (71/151, 47.0%) than in those in the Day 8 Group (61/150, 40.7%). Although delaying the eCG and PGF administration from Day 5 to Day 8 saves one trip through the chute for treatments, it resulted in lower plasma P4 concentrations and tended to decrease pregnancy rates in bovine embryo recipients synchronized with DIB devices and EB and transferred at a fixed time. Furthermore, the administration of injectable P4 at the time of DIB insertion did not affect pregnancy rates.

Treatments with progesterone (P4) releasing devices and estradiol benzoate (EB) have been extensively used in fixed-time AI (FTAI) programs in beef cattle. However, pregnancy rates in postpartum cows kept on pasture often have been lower than expected because of poor body condition score (BCS) and a high incidence of anestrous. A recent study (Cutaia L et al., 2003 Theriogenology 59, 216) suggested that the addition of eCG to a P4/EB FTAI program may improve pregnancy rates in cows with fair to poor BCS, but results were not conclusive. Therefore, this experiment was designed to further investigate the effect of eCG treatment on pregnancy rates in postpartum beef cows in those conditions. The experiment was performed at 2 locations with lactating Angus cows (n=93), 60 to 90d postpartum with a BCS 1.9 (1 to 5 scale; Location 1), and crossbred Zebu cows (n=290), 60 to 90d postpartum with a BCS 2.0 (Location 2). In each Location, cows were randomly allocated to 1 of the following treatment groups: Control, eCG Day6, or eCG Day8. All cows received a P4 intravaginal device (DIB, Syntex, Argentina) and 2mg EB i.m. (Syntex) on Day0, 500mg cloprostenol (Estroplan, Syntex) at the time of DIB removal (Day8), 1mg EB i.m. on Day9 and FTAI 52 to 56h after DIB removal. Cows in the Control group received no further treatment, whereas cows in the eCG-treated groups received 400 IU eCG (Novormon 5000, Syntex) on Day6 or Day8. Cows were examined on Day0 by rectal palpation (Location 1) or by ultrasonography (US; Location 2) and were classified as those with a CL or without a CL, with either large (>8mm) or small (