Short estrous cycles and associated serum progesterone levels in beef cows

Short estrous cycles in beef cows were investigated in two experiments. In trial 1, breeding dates from 2,854 fall-calving Angus cows were used to determine the incidence of short estrous cycles. Of 198 cycles of less than 17 days, 170 were between the first and second detected estrus. Estrous cycles of 7 to 10 days occurred more frequently (P<.005) than other short estrous cycles. Eight-day estrous cycles were most frequent. In the second experiment, calves were weaned from 25 of 33 anestrous spring-calving, crossbred Simmental cows. Compared to herdmates still nursing calves, an increased percentage of cows which had calves weaned were observed in estrus within 10 and 25 days after the date of weaning (P<.005) and a higher percentage had 7- to 10-day estrous cycles. Cows that were observed in estrus within 10 days after weaning were inseminated with Angus semen at their first estrus and with Simmental semen at their second estrus. No cows conceived on the first estrus of 7- to 10-day estrous cycles; 61.5% conceived on the second estrus. Serum progesterone was higher (P<.01) before the second estrus than the first.     

Pregnancy rate of Nelore females inseminated with male-sexed semen

Pregnancy rates of Nelore females inseminated with male-sexed semen and conventional semen from the same bulls were evaluated. The females included 433 heifers (2 years old) and 230 non-suckling cows, totaling 663 animals. Average body condition score was 3.5 (1-5 scale). Estrus was induced with prostaglandin F2α. The total pregnancy rate of females inseminated with male-sexed semen of bulls A, B and C was 38.8% (131/338) less (P<0.0001) than the total pregnancy rate observed for females inseminated with conventional semen from the same bulls (57.9% [188/325]). Pregnancy rates of non-suckling cows inseminated with male-sexed semen was 43.3% (49/113), which was similar (P≥0.05) to the values found for heifers inseminated with male-sexed semen from the same bulls (36.4% [82/225]). The pregnancy rate of females inseminated with male-sexed semen was less compared with females inseminated with conventional semen. In addition, there was no significant difference in the pregnancy rate of heifers versus non-suckling cows.     

Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows

The objective of this study was to compare fixed-time AI pregnancy rate in Angus crossbred beef cows inseminated with frozen-thawed or fresh-extended semen. Two ejaculates from each of two Angus bulls were collected by artificial vagina and pooled for each bull. The pooled semen from each bull was divided into two aliquots; Aliquot 1 was extended using Caprogen^® (LIC, Hamilton, New Zealand) to a concentration of 3 × 10⁶ sperm/straw and Aliquot 2 was extended using egg-yolk-glycerol extender to a concentration of 20 × 10⁶ sperm/straw. Semen extended with Caprogen^® was maintained at ambient temperature and semen extended with egg-yolk-glycerol extender was frozen and maintained at −196 °C until insemination. In each of two breeding seasons (Fall 2007 and Spring 2008), Angus-crossbeef cows (N = 1455) at 12 locations were randomly assigned within location to semen type [Fresh (N = 736) vs. Frozen (N = 719)] and sire [1 (N = 731) vs. 2 (N = 724)]. All cows were synchronized with 100 μg of GnRH im and a progesterone Controlled Internal Drug Release insert (CIDR) on Day 0, and on Day 7, 25 mg of PGF2_α im and CIDR removal. All cows received 100 μg of GnRH im and were inseminated at a fixed-time on Day 10, 66 h after CIDR removal. Timed-AI pregnancy rates were influenced by season (P < 0.05), cows detected in estrus prior to and at AI (P < 0.001), and dam age (P < 0.01). Pregnancy rates were not affected by semen type (Fresh = 51.5% vs. Frozen = 50.4%; P = 0.66) and there were no significant interactions of semen type by estrus expression, semen type by sire, or semen type by season (P > 0.1). In conclusion, commercial beef cows inseminated with fresh-extended semen (3 × 10⁶ sperm/straw) yielded comparable pregnancy rates to conventional frozen-thawed semen in a progesterone supplemented, CO-Synch fixed-time AI synchronization protocol and may provide an alternate to frozen semen for more efficient utilization of superior genetics.     

The effect of addition of equine chorionic gonadotropin to a progesterone-based estrous synchronization protocol in buffaloes (Bubalus bubalis) under tropical conditions

Poor estrus expression and anestrus decrease the reproductive efficiency of buffaloes. The objective of this study was to determine whether the addition of equine chorionic gonadotropin (eCG) to an estrous synchronization protocol and timed insemination could improve ovulation and pregnancy rates of anestrous buffalo cows under tropical conditions. The study population comprised 65 lactating Murrah buffalo cows which were assigned to CIDR (n = 33) or CIDR + eCG (n = 32) treatment groups. Cows in the CIDR group were fitted for 8 d with a controlled intravaginal drug release (CIDR) device containing 1.38 g progesterone, received GnRH (10 μg i.m.) on D 0, PGF_2α (750 μg i.m.) on D 7, and GnRH (10 μg i.m.) on D 9; whereas cows in the CIDR + eCG group received the same treatment plus eCG (500 IU, i.m.) at the time of PGF_2α treatment. All cows were inseminated 16-20 h after the second GnRH treatment. Blood samples were obtained 10 d before the start of synchronization treatment (Day -10) and at the onset of treatment (Day 0). Cows with plasma progesterone concentrations <1 ng/mL recorded in both samples (Low-Low levels of P4) were classified as non-cyclic cows. Similarly, when either one or both of the sample pair contained concentrations of serum progesterone ≥1 ng/mL (High-High, Low-High, or High-Low levels of P4), the buffaloes were classified as cyclic cows. Ovulation rate, defined as the number of buffaloes with at least one corpus luteum 10 days after insemination, was significantly higher (P = 0.018) in the CIDR + eCG (84.4%) cows than in the CIDR cows (57.6%). Pregnancy rate was numerically lower in CIDR (27.3%) than CIDR + eCG (40.6%) cows, though differences were not significant (P = 0.25). Pregnancy rates for CIDR + eCG cows were similar to that of cows inseminated after natural estrus (40.9%; 29/71). In the non-cyclic animals, higher ovulation rates (P = 0.026) were recorded for the CIDR + eCG (81%) than for the CIDR cows (47.4%). Our results indicate that the addition of eCG to a progesterone-based estrous synchronization regimen substantially improves the ovulation rate in non-cyclic buffaloes. When this treatment is followed by timed AI, pregnancy rates achieved in anestrous buffaloes, whether cyclic and non-cyclic, may approach the rates observed in cows inseminated at natural estrus.     

Effect of time interval between prostaglandin F2α and GnRH treatments on occurrence of short estrous cycles in cyclic dairy heifers and cows

Prostaglandin F_2α (PGF_2α) and GnRH treatments, when administered 24 h apart during early diestrus, cause short estrous cycles in some dairy cows and heifers [J. Taponen, M. Kulcsar, T. Katila, L. Katai, G. Huszenicza, H. Rodriguez-Martinez, Short estrous cycles and estrous signs after premature ovulations induced with cloprostenol and gonadotropin-releasing hormone in cyclic dairy cows, Theriogenology 2002; 58, 1291-1302]. We investigated the effect of a time interval between PGF_2α and GnRH administration on the appearance of short cycles. Estrus was induced in heifers with dexcloprostenol. A second luteolysis was induced similarly on day 7 after ovulation, and either 0 (T0) or 24 h (T24) later an injection of GnRH (0.1 mg of gonadorelin) was administered. We monitored ovarian activity with progesterone analyses from blood plasma samples and with ultrasonography. Fourteen cases (12 in T0 and 2 in T24) were excluded due to either incomplete luteolysis (2 cases) or unresponsiveness to GnRH (10 in T0 and 2 in T24). Short estrous cycles (7 to 8 d) were detected in 11/11 and 8/17 heifers in groups T0 and T24, respectively, with a significant difference in the incidence of short cycles (P < 0.01). In Experiment 2, estrus was induced in cows on day 8 (D8, n = 18), 9 (D9, n = 5), or 10 (D10, n = 3) with cloprostenol and gonadorelin administered simultaneously. Daily milk samples were collected for progesterone analysis until subsequent estrus was detected and ovarian ultrasound examinations were performed. Eight cases had to be excluded due to unresponsiveness to GnRH, leaving 18 cases eligible for the study. Short estrous cycles (7-12 d) were detected in 14/18 cows. In conclusion, shortening the time interval between PGF_2α and GnRH treatments increased the incidence of short estrous cycles and appeared to increase the proportion of females unresponsive to GnRH treatment.     

Effects of resynchronization strategies for lactating Holstein cows on pattern of reinsemination, fertility, and economic outcome

The objectives were to evaluate the pattern of re-insemination, pregnancy outcomes to re-insemination in estrus and at fixed time, and economic outcomes of lactating Holstein cows submitted to three resynchronization protocols. Cows were enrolled in the Experiment at 32 ± 3 d after pre-enrollment Artificial Insemination (AI), 7 d before pregnancy diagnosis, and randomly assigned to three resynchronization protocols. All cows diagnosed not pregnant at 39 ± 3 d after pre-enrollment AI were submitted to the Cosynch72 (Day 0 GnRH, Day 7 prostaglandin F_2α, and Day 10 GnRH and fixed time AI). Cows assigned to the control treatment received no further treatment, cows assigned to the GGPG treatment received a GnRH injection on Day −7, and cows assigned to the CIDR treatment received a controlled internal drug release (CIDR) insert containing 1.38 g of progesterone from Days 0-7. Cows observed in estrus were re-inseminated on the same day. Pregnancy was diagnosed at 39 ± 3 and 67 ± 3 d after re-insemination. Costs of the resynchronization protocols were calculated for individual cows enrolled in the study and pregnancies generated were given a value of $275. The GGPG treatment resulted in the slowest (P ≤ 0.06) rate of re-insemination. Overall pregnancy per AI (P/AI) at 39 ± 3 (P = 0.50) and 67 ± 3 (P = 0.49) d after re-insemination were not affected by treatment. Although cost of the control protocol was (P < 0.01) the smallest, return per cow resynchronized was (P < 0.01) greater for GGPG and CIDR protocols. We concluded that presynchronizing the estrous cycle of cows with GnRH or treating cows with a CIDR insert during resynchronization altered the pattern of re-insemination and improved the economic return of resynchronized cows.     

Compared to dinoprost tromethamine, cloprostenol sodium increased rates of estrus detection, conception and pregnancy in lactating dairy cows on a large commercial dairy

Using two PGF_2α treatments 14 days apart as a way to enhance estrus detection rate following the 2nd treatment is a reproductive management tool that continues to be used on large dairy farms. In one study, in cows with a functional CL and a dominant follicle, treatment with cloprostenol vs. dinoprost resulted in greater peripheral estradiol concentrations. The objective of the present study was to determine if cloprostenol could enhance pregnancy rates of cows in a large dairy herd using a PGF_2α program for 1st artificial insemination (AI). Lactating dairy cows (n = 4549) were randomly assigned to receive two treatments of either 500 μg cloprostenol or 25 mg dinoprost 14 days apart, with the 2nd treatment on the 1st day of the voluntary waiting period (57 DIM). Cows detected in estrus within 5 days after the 2nd treatment were inseminated. There was no effect of treatment on day of estrus detection, with 78% of cows inseminated on Days 3 or 4 following treatment. Cloprostenol increased (P < 0.01) estrus detection rates in 1st parity cows compared to dinoprost, 42.4 vs. 34.0%. In cows inseminated on Days 3 or 4 after treatment, cloprostenol increased (P = 0.05) conception rates compared to dinoprost, 38.3 vs. 34.4%. When treatments and parities were combined, conception rates increased (P < 0.02) with interval after treatment (27.0, 36.4, and 44.5% for Days 1 or 2, Days 3 or 4, and Day 5, respectively). Cloprostenol increased (P = 0.02) overall pregnancy rate compared to dinoprost, 14.4 vs. 12.2%. In summary, cloprostenol increased fertility in 1st parity cows inseminated on Days 3 or 4 following treatment and subsequently enhanced pregnancy rates of 1st parity lactating dairy cows compared to dinoprost. Fertility appeared greater in cows expected to have had a young antral ovarian follicle at treatment.     

Luteal competency during the resumption of ovarian cyclicity in postpartum Brahman cows

Twenty pluriparous, spring-calving Brahman cows were used to determine luteal competency, as measured by serum progesterone concentrations, during the first and the second postpartum estrous cycles. Prior to and after calving, all cows were maintained in good body condition on Coastal bermudagrass pasture (IFN 1-00-703). The calves were allowed to suckle ad libitum, and sterile marker bulls were maintained with the cow herd as an aid in estrus detection throughout the trial. Cow weight and body condition score were recorded within 24 hours after calving and again at the first behavioral estrus observed. From day 1 through day 14 (day 0 = estrus) of both the first and the second postpartum estrous cycles, blood samples were collected from each cow, processed to yield serum and analyzed by radioimmunoassay for progesterone concentrations. There was a higher incidence of abnormal estrous cycles following the first postpartum estrus (35%) than following the second (5%) postpartum estrus (P<0.05). The abnormal first estrous cycles were characterized by either a short luteal phase (four cows) or by standing estrus behavior without luteal tissue formation (three cows). When serum progesterone concentrations were compared for all cows during the first estrous cycle with those during the second estrous cycle, there was less progesterone released during the cycle (P<0.05) and lower peak progesterone concentrations (P<0.10) during the first estrous cycle. However, if the abnormal cows were excluded from the analyses, there was no difference (P>0.10) in either progesterone concentrations through the 14 days measured or in peak progesterone concentrations between the first and the second postpartum estrous cycles. It can be concluded from this study that the higher incidence of abnormal luteal function following the first postpartum estrus may contribute to the decreased conception rates observed when cows are bred at their first postpartum estrus.     

Risk factors for resumption of postpartum estrous cycles and embryonic survival in lactating dairy cows

The objectives of this study were to evaluate factors associated with resumption of postpartum estrous cycles and embryonic survival in lactating dairy cows. Holstein cows, 6396 from four dairy farms were evaluated to determine the relationships among parity, body condition score (BCS) at calving and at AI, season of year when cows calved, and milk yield on resumption of postpartum estrous cycles by 65 days postpartum, and all the previous variables, estrual or anestrus and AI protocol on conception rates and embryonic survival at the first postpartum insemination. Cows had their estrous cycle pre-synchronized with two PGF_2α injections given 14 days apart and were inseminated between 69 and 82 days postpartum following either an estrous or ovulation synchronization protocol initiated 12–14 days after the presynchronization. Blood was sampled and analyzed for progesterone twice, 12–14 days apart, to determine whether cows had initiated onset of estrous cycles after calving. Cows were scored for body condition in the week after calving, and again at AI, between 69 and 82 days postpartum. Pregnancy was diagnosed at 30 ± 3 and 58 ± 3 days after AI. Farm influenced all reproductive outcomes evaluated. More (P < 0.0001) multiparous than primiparous cows had initiated estrous cycles. Onset of estrous cycles was also influenced (P < 0.01) by BCS at calving and at AI, BCS change, season, and milk yield. More (P < 0.001) cows that had initiated estrous cycles than anestrous cows were pregnant at 30 and 58 days after AI, but anestrus did not affect pregnancy loss. Conception rates were also influenced (P < 0.01) by parity, BCS at calving and AI, BCS change, and season; however, milk yield and insemination protocol were not associated with conception rates at 30 and 58 days after AI. Factors that reduced conception rate on day 30 after AI also increased pregnancy loss between 30 and 58 days of gestation. Improving BCS at calving and AI, minimizing losses of BCS after calving, and hastening onset of estrous cycles early postpartum are all expected to increase conception because of enhanced embryonic survival.     

Estrus synchronization and pregnancy rates in cyclic and noncyclic beef cows and heifers treated with syncro-mate B or Norgestomet and Alfaprostol

Postpartum lactating cows (N=118) and virgin heifers (N=60) were treated with subcutaneous Norgestomet implants for nine days and received either an intramuscular injection (im) of 5 mg estradiol valerate and 3 mg Norgestomet at the time of implant insertion or an im injection of 5 mg Alfaprostol 24 hr before implant removal. Animals were artificially inseminated 12 hr after detection of estrus. Of the cows and heifers, 78% and 88%, respectively, were in estrus within five days after implant removal (P<0.09). There was no difference between treatments in the proportion of animals in estrus or in the timing of estrus (P<0.85). Estrus was detected in a greater (P<0.05) proportion of animals that were cyclic prior to treatment (88%) than among those that were anestrous prior to treatment (77%). Pregnancy rates after five days were similar between heifers that were cyclic (42%) or anestrous (47%) prior to treatment; however, the five-day pregnancy rate in cows that were anestrous prior to treatment was 38% lower than that in cows that were cyclic prior to treatment (17 vs 55%, P<0.01). Although the treatments synchronized or induced estrus in both cyclic and anestrous animals, marked variability in estrous response and fertility among previously cyclic or anestrous postpartum cows limited the effectiveness of the treatments.     

Evaluation of timed insemination during summer heat stress in lactating dairy cattle

We wished to compare the effect of summer heat stress on pregnancy rate in cows that were inseminated at a set interval associated with a synchronized ovulation vs those inseminated upon routine estrus detection. The study was carried out on a commercial dairy farm in Florida from May to September 1995. Lactating dairy cows were given PGF2 alpha (25 mg i.m.) at 30 + 3 d postpartum and randomly assigned to be inseminated at a set time (Timed group) or when estrus was detected (Control group). Cows in the Timed group were synchronized by sequential administration of Buserelin (8 micrograms i.m.) on Day 0 at 1600 h, PGF2 alpha (25 mg i.m.) on Day 7 at 1600 h and Buserelin (8 micrograms i.m.) on Day 9 at 1600 h. They were inseminated on Day 10 between 0800 and 0900 h (Day 9 + 16 h). Cows in the Control group were given PGF2 alpha at 57 + 3 d postpartum and inseminated when detected in estrus. Estrus detection or insemination rate for control insemination cows was 18.1 +/- 2.5% versus 100% for time inseminated cows (P < 0.01). Mean interval from PGF2 alpha to insemination was shorter for time inseminated cows (3 +/- 2.1 d < 35.5 +/- 1.9 d; P < 0.01). Pregnancy rate was greater for time inseminated cows (13.9 +/- 2.6 > 4.8 +/- 2.5%; P < 0.01) as was overall pregnancy rate by 120 d postpartum (27.0 +/- 3.6 > 16.5 +/- 3.5%; P < 0.05). Number of days open for cows conceiving by 120 d postpartum was less for time inseminated cows (77.6 +/- 3.8 < 90.0 +/- 4.2 d; P < 0.05), as was interval to first service (58.7 +/- 2.1 < 91.0 +/- 1.9 d; P < 0.01). Services per conception were greater for time inseminated cows (1.63 +/- 0.10 > 1.27 +/- 0.11; P < 0.05). The timed insemination program did improve group reproductive performance. However, the timed insemination program will not protect the embryo from temperature-induced embryonic mortality, but management limitations induced by heat stress on estrus detection are eliminated. An economical evaluation of the timed insemination program indicates an increase in net revenue per cow with implementation of timed insemination for first service during the summer months.     

Timed artificial insemination plus heat I: effect of estrus expression scores on pregnancy of cows subjected to progesterone–estradiol-based protocols

Expression of estrus near timed artificial insemination (TAI) is associated with greater fertility, and estrus detection could improve TAI fertility or direct TAI management, although accurate estrus detection can be difficult and time-consuming using traditional methods. The aim of this study is to evaluate influence of estrus on pregnancy (artificial insemination pregnancy rates (P/AI)) and to validate an alternative method to classify estrus/heat expression using tail chalking (HEATSC) in postpartum Bos indicus cows subjected to TAI in progesterone–estrogen-based protocols. In experiment 1 (Exp. 1), cows (5491) were subjected to visual observation of estrus after progesterone device removal, before TAI, and P/AI was evaluated according to estrus and body condition score (BCS). Cows received a progesterone device and 2 mg estradiol benzoate (EB). After 8 days, the device was removed and 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotrophin was given. Later, animals in Exp. 1 received 1 mg EB and TAI 44 to 48 h. In the Exp. 2 – 3830 cows using similar protocol, received different ovulation inducers: 1 mg EB (n=1624) or 1 mg estradiol cypionate (EC; n=2206) on day 8 (D8). Cows were then marked with chalk, and HEATSC evaluated at TAI on D10 (HEATSC1 – no chalk removal=no estrus expression; HEATSC2 – partial chalk removal=low estrus expression; HEATSC3 – near complete/complete chalk removal=high estrus expression). In Exp. 1, cows showing estrus presented greater P/AI (48.4% v. 40.2%, P<0.05). In Exp. 2, P/AI (HEATSC1 – 40.0%; HEATSC2 – 49.7%; HEATSC3 – 60.9%; P<0.001), and larger follicle timed artificial insemination (LFTAI) (<0.001) varied according to HEATSC. There was no difference in P/AI (P=0.41) or LFTAI (P=0.33) according to ovulation inducer. Cows with greater BCS showed greater P/AI in both experiments (P<0.05). Estrus presence and greater HEATSC improved P/AI, and EC v. EB used promoted differential estrus manifestation (cows showing HEATSC2 and HEATSC3: 79.5% with EB v. 69.98% with EC use, P<0.001), however, with similar P/AI. The use of HEATSC in B. indicus cows subjected to TAI is useful to identify cows with greater estrus expression and consequently improved pregnancy rates in TAI, allowing the cows with low HEATSC to be targeted for additional treatments aimed at improving P/AI.     

Effect of estrous cycle stage at Syncro-Mate B treatment on conception and time to estrus in cattle

Two trials involving 85 heifers and 67 cows were conducted to determine the effect of estrous cycle stage at the time of Syncro-Mate-B((R)) (SMB) treatment on interval to estrus following implant removal and on conception rate at the synchronized estrus. In Trial 1, 57 beef and 28 dairy heifers were treated with SMB on each representative day of a 22-d estrous cycle (estrus = Day 0). Beef heifers were artificially inseminated approximately 48 h after implant removal, whereas dairy heifers were inseminated 0 to 12 h after detection of estrus. Inseminations were scored by the inseminator according to their difficulty. Interval to the onset of estrus was not different between heifers treated early ( Day 11) in the cycle (35.2 +/- 7.2 h). Conception rate at the synchronized estrus was slightly higher in early-cycle heifers (22 47 = 47% ) compared to late-cycle heifers (14 38 = 37% , P = 0.2). Heifers that were difficult to inseminate had lower (P < 0.01) conception rates (2 11 = 18% ) at the synchronized estrus than heifers considered normal (21 51 = 41% ) or easier than normal to inseminate (13 23 = 57% ). In Trial 2, of the 131 beef cows synchronized, 67 that were estimated to be either early or late in the estrous cycle by progesterone analysis were utilized. Cows were treated with SMB and inseminated without regard to estrus 48-h after implant removal. Inseminations were scored as in Trial 1. Calves were separated from cows from the time of implant removal to insemination. Conception rate was higher (P < 0.05) in cows treated with SMB early ( Day 11, 16 35 = 46% ). Cows that were difficult to inseminate had a lower (P < 0.01) conception rate (0 8 = 0% ) than cows that were normal (43 94 = 46% ) or easier than normal to inseminate (13 29 = 45% ).     

Estrous expression during a fixed-time artificial insemination protocol enhances development and interferon-tau messenger RNA expression in conceptuses from Bos indicus beef cows

Expression of estrus (EST) near the time of fixed-time artificial insemination (FTAI) increases pregnancy success in beef females. This outcome has been associated with improved pregnancy establishment and maintenance, although research is still warranted to validate this theory. Hence, this experiment compared ovarian, uterine and conceptus factors associated with pregnancy establishment in Bos indicus beef cows according to estrous expression during a FTAI protocol. One hundred lactating multiparous Nelore cows received a 2 mg injection of estradiol benzoate and an intravaginal progesterone (P4) releasing device on day −11, a 12.5 mg injection of prostaglandin F_2α on day −4, P4 device removal in addition to 0.6 mg injection of estradiol cypionate and 300 IU injection of equine chorionic gonadotropin on day −2, and FTAI on day 0. An estrous detection patch was attached to the tailhead of each cow on day −2, and estrous expression was defined as removal of >50% of the rub-off coating from the patch at FTAI. Overall, 39 cows expressed EST, 55 did not express EST (NOEST), and six cows lost their patch and were discarded from the experiment. Ovarian ultrasonography was performed at FTAI, and on days 7 and 15 of the experiment. Blood samples were also collected on days 7 and 15. Only cows without a corpus luteum (CL) on day 0, and with a CL on days 7 and 15 remained in the experiment (EST, n=36; NOEST, n=48). On day 15, cows were randomly selected within each group (EST, n=29; NOEST, n=30) for conceptus collection via transcervical flushing, followed by endometrial biopsy in the uterine horn ipsilateral to the CL. Within cows not assigned to conceptus collection, blood samples were collected for whole blood RNA extraction (day 20) and pregnancy status was verified by transrectal ultrasonography (day 30). Diameter of dominant follicle on day 0 and plasma P4 concentrations on day 7 were greater (P⩽0.02) in EST v. NOEST cows. Conceptus length and messenger RNA (mRNA) expression of prostaglandin E synthase and interferon-tau were greater (P⩽0.04) in EST v. NOEST cows. Moreover, EST cows diagnosed as pregnant on day 30 had greater (P<0.01) blood mRNA expression of myxovirus resistance 2 on day 20 compared with NOEST. In summary, estrous expression near the time of FTAI enhanced pregnancy establishment factors in B. indicus cows, including conceptus development and mRNA expression of interferon-tau.     

Comparison of long-term CIDR-based protocols to synchronize estrus in beef heifers

Two experiments evaluated long-term controlled internal drug release (CIDR) insert-based protocols to synchronize estrus and compare differences in their potential ability to facilitate fixed-time artificial insemination (FTAI) in beef heifers. In Experiment 1 estrous cycling heifers (n = 85) were assigned to one of two treatments by age and body weight (BW). Heifers with T1 received a CIDR from days 0 to 14, gonadotropin releasing hormone (GnRH) on day 23, and prostaglandin F_2α (PG) on day 30. Heifers with T2 received a CIDR from days 2 to 16, GnRH on day 23, and PG on day 30. Ovaries were evaluated by ultrasonography on days 23 and 25 to determine ovulatory response to GnRH. In Experiment 2 heifers (n = 353) were assigned within reproductive tract scores by age and BW to one of four treatments. Heifers in T1 and T2 received the same treatments described in Experiment 1. Heifers in T3 and T4 received the same treatments as T1 and T2, respectively, minus the addition of GnRH. In Experiments 1 and 2, heifers were fitted with HeatWatch transmitters for estrous detection and AI was performed 12 h after estrus. In Experiment 1 heifers assigned to T1 had larger dominant follicles at GnRH compared to T2 (P < 0.01) but response to GnRH, estrous response after PG, mean interval to estrus, and variance for interval to estrus after PG did not differ (P > 0.10). AI conception and final pregnancy rate were similar (P > 0.50). In Experiment 2 estrous response after PG did not differ (P > 0.70). Differences in mean interval to estrus and variance for interval to estrus (P < 0.05) differed based on the three-way interaction of treatment length, GnRH, and estrous cyclicity status. AI conception and final pregnancy rates were similar (P > 0.10). In summary, the greater estrous response following PG and resulting AI conception and final pregnancy rates reported for heifers assigned to the two treatments in Experiment 1 and among the four treatments in Experiment 2 suggest that each of these long-term CIDR-based protocols was effective in synchronizing estrus in prepubertal and estrous cycling beef heifers. However, the three-way interaction involving treatment length, GnRH, and estrous cyclicity status in Experiment 2 clearly suggests that further evaluation of long-term CIDR-based protocols is required with and without the addition of GnRH and on the basis of estrous cyclicity status to determine the efficacy of these protocols for use in facilitating FTAI.     

Timed artificial insemination plus heat II: gonadorelin injection in cows with low estrus expression scores increased pregnancy in progesterone/estradiol-based protocol

The use of tail chalk and estrus/heat expression scores (HEATSC) evaluation is instrumental in identifying cows with greater estrus expression and greater artificial insemination pregnancy rates (P/AI) in cows submitted to timed artificial insemination (TAI), and cows with low or no estrus expression present lower P/AI. It was intended in this study to improve the pregnancy rates in TAI for Bos indicus beef cows, and gonadotrophin-releasing hormone (GnRH) injection was hypothesized to increase pregnancy rates in a TAI program for cows submitted to progesterone–estradiol-based protocols with low or no estrus expression, evaluated by HEATSC. Cows (n= 2284) received a progesterone device and 2 mg estradiol benzoate, after 8 days the device was removed and 1 mg estradiol cypionate, 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotropin was administered. All cows were marked with chalk and HEATSC evaluated (scales 1 to 3) at TAI performed on day 10. Animals with HEATSC1 and HEATSC2 (n= 937) received 100 μg de gonadorelin (GNRH group; n= 470), or 1 ml saline (Control group; n= 467), and cows with HEATSC3 (named HEAT group; n= 1347) received no additional treatment. The larger dominant follicle, evaluated on day 8and at TAI (day 10), was greater in HEAT group (P= 0.0145 and P <0.001, respectively). Corpus luteum (CL) area and progesterone concentration was evaluated on day 17, and CL area was larger in HEAT group, intermediary in Control and lower in GnRH group (Control= 2.68 cm², GnRH= 2.37 cm², HEAT group= 3.07 cm², P <0.001). Greater progesterone concentrations were found in HEAT group than in Control and GnRH groups (Control= 4.74 ng/ml, GnRH= 4.29 ng/ml, HEAT group= 6.08 ng/ml, P<0.001). There was a difference in ovulation rate, greater in HEAT group than GnRH and Control groups (Control= 72.5%; GnRH= 81.25%; HEAT group= 90.71%; P= 0.0024). Artificial insemination pregnancy rates was greater in HEAT group (57.09% (769/1347) than in Control and GNRH groups, with positive effect of GnRH injection at the time of TAI in P/AI (Control= 36.18% (169/467), GnRH= 45.95% (216/470); P<0.0001). In conclusion, GnRH application in cows with low HEATSC (1 and 2) is a simple strategy, requiring no changes in TAI management to increase pregnancy rates in postpartum beef cows submitted to progesterone–estradiol-based TAI protocols, without reaching, however, the pregnancy rates of cows that demonstrate high estrus expression at the TAI.     

Breeding capacity, behavior and fertility of bulls with Brahman genetic influence during synchronized breeding of beef females

The objective of this experiment was to evaluate the feasibility of using bulls of Brahman-derived breeds for synchronized breeding of females treated with Syncro-Mate-B (SMB). Suckled, postpartum cows (n = 261) and virgin heifers (n = 227) were given the standard SMB treatment. Calves were removed from cows for 48 h beginning at implant removal, and individual bulls were placed in pens with 15 to 20 females (x = 1:15.7 ) for 48 h beginning at implant removal. Bulls (n = 31) were 18 to 36 mo of age and had scored satisfactorily on a breeding soundness evaluation (BSE). The mean +/- SEM percentage of females in estrus, percentage of estrous females serviced and percentage of total females serviced were 77.2 +/- 2.4, 72.9 +/- 2.9 and 55.7 +/- 3%, respectively. Total average services per bull was 23.6 +/- 1.6; however, average number of individuals serviced was only 8.6 +/- 0.42. Mean +/- SEM percentage of females conceiving of those serviced, percentage conceiving of those in estrus and percentage conceiving of total females available were 57.3 +/- 3.7, 40.6 +/- 2.6 and 32.6 +/- 2.5%, respectively. Mean BSE scores of eight Simbrah bulls tested immediately before and 5 d after synchronized breeding did not differ (P > 0.05). These data suggest that the conception rate of Brahman-influenced bulls servicing SMB-synchronized cows is within the normal range. However, the total number of individual females serviced and the total pregnancy rate under the protocol employed was low. This occurred because all estrous females were not inseminated, and some bulls exhibited unpredicted low fertility.     

Luteal function,largest follicle,and fertility in postpartum dairy cows treated with 14dCIDR-PGF2α versus 2xPGF2α-Ovsynch for timed AI

A method for timed artificial insemination (AI) that is used for beef cows, beef heifers, and dairy heifers employs progesterone-releasing inserts, such as the controlled internal drug release (CIDR; Zoetis, New York, NY, USA) that are left in place for 14 days. The 14-day CIDR treatment is a method of presynchronization that ensures that cattle are in the late luteal phase of the estrous cycle when PGF_2α is administered before timed AI. The objective of this study was to test the effectiveness of the 14dCIDR-PGF_2α program in postpartum dairy cows by comparing it with the traditional “Presynch-Ovsynch” (2xPGF_2α-Ovsynch) program. The 14dCIDR-PGF_2α cows (n = 132) were treated with a CIDR insert on Day 0 for 14 days. At 19 days after CIDR removal (Day 33), the cows were treated with a luteolytic dose of PGF_2α, 56 hours later were treated with an ovulatory dose of GnRH (Day 35), and 16 hours later were inseminated. The 2xPGF_2α-Ovsynch cows were treated with a luteolytic dose of PGF_2α on Day 0 and again on Day 14. At 12 days after the second PGF_2α treatment (Day 26), the cows were treated with GnRH. At 7 days after GnRH, the cows were treated with PGF_2α (Day 33), then 56 hours later treated with GnRH (Day 35), and then 16 hours later were inseminated. There was no effect of treatment or treatment by parity interaction on pregnancies per AI (P/AI) when pregnancy diagnosis was performed on Day 32 (115/263; 43.7%) or Days 60 to 90 (99/263; 37.6%) after insemination. There was an effect of parity (P < 0.05) on P/AI because primiparous cows had lesser P/AI (35/98; 35.7%) than multiparous cows (80/165; 48.5%) on Day 32. Cows observed in estrus after the presynchronization step (within 5 days after CIDR removal or within 5 days after the second PGF_2α treatment) had greater P/AI than those not observed in estrus (55/103; 53.4% vs. 60/160; 37.5%; observed vs. not observed; P < 0.01; d 32 pregnancy diagnosis). When progesterone data were examined in a subset of cows (n = 208), 55.3% of cows had a “prototypical” response to treatment (i.e., the cow had an estrous cycle that was synchronized by the presynchronization treatment and then the cow responded appropriately to the subsequent PGF_2α and GnRH treatments before timed AI). Collectively, cows with a prototypical response to either treatment had 52.2% P/AI that was greater (P < 0.001) than the P/AI for cows that had a nonprototypical response (19%) (P/AI determined at 60–90 days of pregnancy). In conclusion, we did not detect a difference in P/AI when postpartum dairy cows were treated with 14dCIDR-PGF_2α or 2xPGF2α-Ovsynch before timed AI. The primary limitation to the success of either program was the failure of the cow to respond appropriately to the sequence of treatments.     

Association between leptin single nucleotide polymorphism and reproductive performance of lactating Holstein cows

The objectives of the current study were to evaluate the associations among reproductive performance of Holstein cows, single nucleotide polymorphism (SNP) in the R4C locus of the Exon-2 region of the leptin gene and health disorders. Lactating dairy cows (n=814) had their DNA sequenced at the Exon-2 region of the leptin gene to determine the presence of SNP in the R4C locus. Cows had the stage of estrous cycle synchronized with two injections of PGF(2α) 14d apart, with or without a progesterone insert. After the second PGF(2α), cows were either inseminated on detected estrus and, if not inseminated within 13d, cows were submitted to a timed artificial insemination (AI) protocol 13d after the second PGF(2α). Blood was sampled at 35±3, 49±3, and 62±3d in milk (DIM) and cows with progesterone <1.0ng/mL in the first two samples were considered to be anovular, whereas those with at least one of the first two samples with progesterone ≥1.0ng/mL were considered to have initiated estrous cyclicity. Anovular cows with progesterone ≥1.0ng/mL at 62±3DIM were considered to have resumed estrous cyclicity. Pregnancy was diagnosed at 31 and 60d after first postpartum AI and at 42d after subsequent inseminations. Resulting genotypes were CC (34.6%), CT (48.2%), and TT (17.2%). Leptin genotype was associated (P=0.03) with increased prevalence of estrous cyclic cows at 49±3DIM, as TT cows were and tended to be less likely to be estrous cyclic than CC and CT cows, respectively. Other measures of reproductive performance in the first 305DIM were not associated with leptin genotype. Cows diagnosed with at least one postparturient disease were less likely to become pregnant after first (P<0.01) and second (P=0.02) AI and to be pregnant at 305DIM (P<0.01). Furthermore, cows diagnosed with a disease event had a 36% reduction (P<0.001) in the rate of pregnancy, which extended the median interval to pregnancy by 60d. Estrous cyclic cows had a 35% increased rate of pregnancy, which reduced the interval to pregnancy by 34d. Cows homozygous for the T allele were less likely to be cyclic early postpartum, but leptin genotype was not associated with other reproductive responses when cows were subjected to a controlled breeding program.     

Reproductive performance of lactating dairy cows treated with cloprostenol, hcg and estradiol benzoate for synchronization of estrus followed by timed AI

In previous studies, we demonstrated that the administration of a luteolytic dose of cloprostenol, followed by 750 IU hCG plus 3 mg estradiol benzoate (EB) 12 h later, synchronized estrus in cows in the luteal phase. Most cows were ready for service 48 h after the beginning of treatment. The objectives of this study were to evaluate the reproductive performance of lactating dairy cows treated with this method of estrus synchronization and to determine the effect of decreasing the hCG-EB dose on synchronization and pregnancy rates after timed AI. Data were obtained from cows first inseminated within an interval of 45 to 70 d postpartum. A total of 2,472 lactating dairy cows in their first to second lactation period were assigned to 4 groups. Cows estimated to be in the luteal phase by rectal palpation were treated with 500 mcg, im, of cloprostenol and assigned to 1 of 3 groups to be intramuscularly injected with hCG-EB 12 h later at the following doses: Group 1 (n=626), 250 IU of hCG and 1 mg of EB; Group 2 (n=592), 500 IU of hCG and 2 mg of EB; and Group 3 (n=664), 750 IU of hCG and 3 mg of EB. Cows displaying natural estrus were inseminated to serve as controls (n=590). The synchronized cows were inseminated 48 h after cloprostenol injection, and control animals visually determined to be in natural estrus during the morning or afternoon were inseminated the following morning. Pregnancy diagnosis was performed by rectal palpation at 34 to 40 d postinsemination. All synchronized cows showed estrous activity within 24 to 36 h after cloprostenol treatment and were considered to be ready for service 48 h after this treatment. There was a significant effect of treatment on the pregnancy rate, either to first AI or to 2 rounds of AI. The pregnancy rate in response to first or second rounds of AI was similar to control rates for cows in Groups 1 and 2, and lower than control rates in Group 3. Cows in Group 1 showed a higher pregnancy rate to first AI than those in Group 3 (P<0.0001), and a higher pregnancy rate to second AI rounds than cows in Groups 2 (P<0.02) and 3 (P<0.0001). The number of cows returning to estrus was unaffected by treatment. However, treatment significantly decreased (P<0.01) the time of return to estrus as the hCG-EB dose increased. These findings indicate that the lowest dose of hCG-EB treatment tested gave the overall best pregnancy results among the treated groups. Furthermore, the synchronization protocol used in this experiment allows effective AI management of lactating dairy cows without the need for estrus detection.