Improved conception in timed-artificial insemination using a progesterone-releasing intravaginal device and Ovsynch protocol in postpartum suckled Japanese Black beef cows

The primary objective was to determine the effect of supplemental progesterone, administered via an intravaginal device (CIDR), on conception rates to timed-artificial insemination (timed-AI) in postpartum suckled Japanese Black beef cows treated with the Ovsynch protocol. A secondary objective was to compare the effects of treatments on plasma concentrations of progesterone and estradiol. Cows in the control group (Ovsynch, n=38) received a standard Ovsynch protocol (100 microg GnRH analogue on Day 0, 500 microg PGF2alpha analogue on Day 7, and 100 microg GnRH analogue on Day 9), with AI on Day 10, approximately 20 h after the second GnRH treatment. Cows in the treatment group (Ovsynch+CIDR; n=40) received a standard Ovsynch protocol plus a CIDR for 7 days (starting on Day 0). Plasma progesterone concentrations were determined on Days 0, 1, 7, 9, 10, and 17 and plasma estradiol-17beta concentrations were determined on Days 7, 9, 10, and 17. The odds ratio for likelihood of conception was 3.29 times greater (P=0.02) in the Ovsynch+CIDR group compared to Ovsynch group. The conception rate was greater (P=0.03) in the Ovsynch+CIDR group than in the Ovsynch group (72.5% versus 47.7%). Insertion of a CIDR device significantly increased plasma progesterone concentrations only on Days 1 and 7 (P<0.001 and P=0.05, respectively), but had no significant effect on plasma estradiol-17beta concentrations. Including a CIDR with the Ovsynch protocol significantly improved conception rates in postpartum suckled Japanese Black beef cows.     

Effect of CIDR-based protocols for timed-AI on the conception rate and ovarian functions of Japanese Black beef cows in the early postpartum period

Our objectives were to compare: (1) conception rates (in early postpartum Japanese Black beef cows) to timed-artificial insemination (timed-AI) among Ovsynch and Ovsynch plus CIDR protocols, and a protocol that used estradiol benzoate (EB) in lieu of the first GnRH of the Ovsynch plus CIDR; and (2) the effects of these protocols on blood concentrations of ovarian steroids. Cows in the control group (Ovsynch; n=35) underwent a standard Ovsynch protocol (GnRH analogue on Day 0, PGF(2 alpha) analogue on Day 7 and GnRH analogue on Day 9), with timed-AI on Day 10, approximately 20 h after the second GnRH treatment. Cows in the Ovsynch+CIDR group (n=31) received a standard Ovsynch protocol plus a CIDR for 7 days (starting on Day 0). Cows in the third treatment group (EB+CIDR+GnRH; n=41) received 2mg of EB on Day 0 in lieu of the first GnRH treatment, followed by the same treatment as in the Ovsynch+CIDR protocol. The conception rate tended to be greater in the Ovsynch+CIDR group (67.7%, P<0.15) and was greater in the EB+CIDR+GnRH (73.2%, P<0.05) and CIDR-combined (both CIDR-treated groups were combined) groups (70.8%, P<0.05) than in the Ovsynch group (48.6%). Plasma progesterone concentrations were higher on Day 7 (P<0.01) and lower on Days 14, 17 and 21 (P<0.001) in the CIDR-combined group than in the Ovsynch group. Plasma estradiol-17beta concentrations were higher on Day 7 in the Ovsynch group of non-pregnant cows than in the CIDR-combined group of non-pregnant cows and in an all-combined group (all treatment groups combined) of pregnant cows (P<0.01). Furthermore, estradiol-17beta concentrations were lower on Day 9 in the Ovsynch and CIDR-combined groups of non-pregnant cows than in the all-combined group of pregnant cows (P<0.05). In conclusion, both protocols using CIDR improved conception rates following timed-AI in early postpartum suckled Japanese Black beef cows relative to the Ovsynch protocol. Treatment with a CIDR may prevent early maturation of follicles observed in non-pregnant cows treated with the Ovsynch protocol, by maintaining elevated blood progesterone concentrations until PGF(2 alpha) treatment.     

Synchronization rate, size of the ovulatory follicle, and pregnancy rate after synchronization of ovulation beginning on different days of the estrous cycle in lactating dairy cows

Recently a protocol was developed that precisely synchronizes the time of ovulation in lactating dairy cows (Ovsynch; GnRH-7d-PGF2 alpha-2d-GnRH). We evaluated whether initiation of Ovsynch on different days of the estrous cycle altered the effectiveness of this protocol. The percentage of cows (n = 156) ovulating to the first GnRH was 64% and varied (P < 0.01) by stage of estrous cycle. Treatment with PGF2 alpha was effective, with 93% of cows having low progesterone at second GnRH. The overall percentage of cows that ovulated after second GnRH (synchronization rate) was 87% and varied by response to first GnRH (92% if ovulation to first GnRH vs 79% if no ovulation; P < 0.05). There were 6% of cows that ovulated before the second injection of GnRH and 7% with no detectable ovulation by 48 h after second GnRH. Maximal diameter of the ovulatory follicle varied by stage of estrous cycle, with cows in which Ovsynch was initiated at midcycle having the smallest follicles. In addition, milk production and serum progesterone concentration on the day of PGF2 alpha affected (P < 0.05) size of the ovulatory follicle. Using these results we analyzed pregnancy rate at Days 28 and 98 after AI for cows (n = 404) in which Ovsynch was initiated on known days of the estrous cycle. Pregnancy rate was lower for cows expected to ovulate larger follicles than those expected to ovulate smaller follicles (P < 0.05; 32 vs 42%). Thus, although overall synchronization rate with Ovsynch was above 85%, there were clear differences in response according to day of protocol initiation. Cows in which Ovsynch was initiated near midcycle had smaller ovulatory follicles and greater pregnancy rates.     

Luteal activity at the onset of a timed insemination protocol affects reproductive outcome in early postpartum dairy cows

This study was designed to compare two timed insemination protocols, in which progesterone, GnRH and PGF2alpha were combined, with the Ovsynch protocol in presynchronized, early postpartum dairy cows. Reproductive performance was also evaluated according to whether cows showed high or low plasma progesterone concentration, at the onset of treatment. One hundred and six early postpartum dairy cows were presynchronized with two cloprostenol treatments given 14 days apart, and then assigned to one of the three treatment groups. Treatments for the synchronization of estrus in all three groups started 7 days after the second cloprostenol injection, which was considered Day 0 of the actual treatment regime. Cows in the control group (Ovsynch, n=30) were treated with GnRH on Day 0, PGF2alpha on Day 7, and were given a second dose of GnRH 32 h later. Cows in group PRID (n=45) were fitted with a progesterone releasing intravaginal device (PRID) for 9 days, and were given GnRH at the time of PRID insertion and PGF2alpha on Day 7. In group PRID/GnRH (n=31), cows received the same treatment as in the PRID group, but were given an additional GnRH injection 36 h after PRID removal. Cows were inseminated 16-20 h after the administration of the second GnRH dose in the Ovsynch group, and 56 h after PRID removal in the PRID and PRID/GnRH groups. Ovulation rate was determined on Day 11 postinsemination by detecting the presence of a corpus luteum in the ovaries. Lactation number, milk production, body condition at the onset of treatment and treatment regime were included as potential factors influencing ovulation and pregnancy after synchronization. Logistic regression analysis for cows with high and low progesterone concentration on treatment Day 0 revealed that none of the factors included in the models, except the interaction between progesterone and treatment regime, influenced the risk of ovulation and pregnancy significantly. In cows with high progesterone concentration at treatment onset, Ovsynch treatment resulted in a significantly improved pregnancy rate over values obtained following PRID or PRID/GnRH treatment. In cows with low progesterone concentration, PRID or PRID/GnRH treatment led to markedly increased ovulation and pregnancy rates with respect to Ovsynch treatment. These findings suggest the importance of establishing ovarian status in early postpartum dairy cows before starting a timed AI protocol, in terms of luteal activity assessed by blood progesterone.     

Effect of an implant containing the GnRH agonist deslorelin on secretion of LH, ovarian activity and milk yield of postpartum dairy cows

Prevention of high plasma progesterone concentrations in the early postpartum period may improve fertility. Our objective was to determine whether a Deslorelin implant (DESL; 2100 microg, s.c.) would reduce secretion of LH and alter follicle dynamics, plasma concentrations of progesterone, estradiol and PGF2alpha metabolite (PGFM) in postpartum dairy cows. Cows received DESL on Day 7 postpartum (Day 7, n=8) or were untreated (Control, n=9). All cows were injected with GnRH (100 microg, i.m.) on Day 14 to assess LH response. A protocol for synchronization of ovulation with timed AI was initiated on Day 60 (GnRH [Day 60], CIDR [Day 60 to Day 67], PGF2alpha [Day 67, 25 mg and Day 68, 15 mg], GnRH [Day 69] , AI [Day 70]). The LH response to injection of GnRH on Day 14 was blocked in animals treated with DESL. Numbers of Class 1 (<6 mm) follicles were unaffected (P > 0.05) whereas numbers of Class 2 (6 to 9 mm) (P < 0.01) and Class 3 (>9 mm) follicles were less (P < 0.01) in DESL cows between Day 7 and Day 21. From Day 22 to Day 60, DESL-treated cows had more of Class 1 follicles and less Class 2 (P < 0.01) and Class 3 (P < 0.01) follicles, and lower plasma concentrations of progesterone and estradiol (P < 0.01). Concentrations of PGFM between Day 7 and Day 42 were not affected by treatment (P > 0.05). All cows ovulated in response to GnRH on Day 69. Subsequent luteal phase increases in plasma progesterone concentrations (Day 70 to Day 84) did not differ. The use of the DESL implant associated with PGF2alpha given 14 days later suppressed ovarian activity and caused plasma progesterone concentrations to remain < 1 ng/mL between Day 22 and Day 51. The DESL implant did not affect milk production.     

Strategic treatment of anovular dairy cows with GnRH

The primary objective was to evaluate fertility of anovular dairy cows given GnRH 4 d after first postpartum timed AI (TAI). Secondary objectives were to determine ovulatory response to treatment, effect of treatment on serum progesterone (P(4)) concentrations, and the proportion with a short luteal phase. Lactating Holstein cows (n=1047) were submitted for first postpartum TAI using a Presynch+Ovsynch protocol. Anovular cows were identified from an initial 1047 lactating Holstein cows using transrectal ultrasonography, based on the absence of a CL at the first GnRH injection of a Presynch+Ovsynch protocol, and anovular cows were randomly assigned to receive either no further treatment (Control, n=85), or 100 microg of GnRH 4 d after TAI (GnRH treated; n=71). For GnRH treated cows, 51% responded by ovulating a follicle in response to GnRH treatment 4 d after TAI; however, pregnancies per AI (P/AI) did not differ between GnRH treated cows that ovulated (36%) compared to GnRH treated cows that did not ovulate (21%). There was a quadratic effect of P(4) at the PGF(2 alpha) injection of Ovsynch on P/AI, and cows with P(4)>or=1 ng/mL at the PGF(2 alpha) injection of Ovsynch had greater P/AI (41%) than cows with P(4)<1 ng/mL (12%); however, no treatment difference was detected. Overall, P/AI did not differ between control (30.1%) and GnRH treated (29.6%) treatments for synchronized cows. Although treatment of anovular cows with GnRH 4 d after TAI failed to improve fertility, variation among cows in serum P(4) at the PGF(2 alpha) injection of Ovsynch dramatically affected fertility of anovular dairy cows.     

The use of progestins in regimens for fixed-time artificial insemination in beef cattle

Four experiments were conducted to investigate modifications to gonadotropin releasing hormone (GnRH)-based fixed-time Al protocols in beef cattle. In Experiment 1, the effect of reducing the interval from GnRH treatment to prostaglandin (PGF) was examined. Lactating beef cows (n = 111) were given 100 mg gonadorelin (GnRH) on Day 0 (start of treatment) and either 500 microg cloprostenol (PGF) on Day 6 with Al and 100 microg GnRH 60 h later, or PGF on Day 7 with Al and GnRH 48 h later (6- or 7-day Co-Synch regimens). Pregnancy rates were 32/61 (53.3%) versus 26/50 (52.0%), respectively (P = 0.96). In Experiment 2. cattle (n = 196) were synchronized with a 7-day Co-Synch regimen and received either no further treatment or a CIDR-B device (Days 0-7). Pregnancy rates were 32/71 (45.1%) versus 33/77 (42.9%) in cows (P < 0.8), and 9/23 (39.1 %) versus 17/25 (68.0%) in heifers (P < 0.05). In Experiment 3, 49 beef heifers were randomly assigned to receive 12.5 mg pLH on Day 0, PGF on Day 7 and 12.5 mg of pLH on Day 9 with Al 12 h later (pLH Ovsynch), or similar treatment plus a CIDR-B device from Days 0 to 7 (pLH Ovsynch + CIDR-B), or 1 mg estradiol benzoate (EB) and 100 mg progesterone on Day 0, a CIDR-B device from Days 0 to 7 (EB/ P4 + CIDR-B), PGF on Day 7 (at the time of CIDR-B removal) and 1 mg i.m. EB on Day 8 with AI on Day 9 (52 h after PGF). Pregnancy rate in the EB/P4 + CIDR-B group (75.0%) was higher (P < 0.04) than in the pLH Ovsynch group (37.5%): the pLH Ovsynch + CIDR-B group was intermediate (64.7%). In Experiment 4, 266 non-lactating cows were allocated to a 7-day Co-Synch protocol (Co-Synch), a 7-day Co-Synch plus 0.6 mg per head per day melengestrol acetate (MGA) from Days 0 to 6 inclusive (Co-Synch + MGA) or MGA (Days 0-6) plus 2 mg EB and 50 mg progesterone on Day 0. 500 microg PGF on Day 7, 1 mg EB on Day 8 and fixed-time Al 28 h later (EB/ P4 + MGA). Pregnancy rates (P < 0.25) were 44.8% (39/87: Co-Synch), 47.8% (43/90; Co-Synch + MGA), and 60.7% (54/89: EB/P4 + MGA). In conclusion, a 6- or 7-day interval from GnRH to PGF in a Co-Synch regimen resulted in similar pregnancy rates in cows. The addition of a progestin to a Co-Synch or Ovsynch regimen significantly improved pregnancy rates in heifers but not in cows. Progestin-based regimens that included EB consistently resulted in high pregnancy rates to fixed-time Al.     

Is Doublesynch protocol a new alternative for timed artificial insemination in anestrous dairy cows

This is the very first report that suggests high pregnancy rates can be obtained with use of the Doublesynch protocol in anestrous dairy cows. Recently, a new synchronization method has been developed (Doublesynch) that resulted in synchronized ovulations both after the first and second gonadotropin-releasing hormone (GnRH) treatments. It was suggested that this protocol has the potential to increase the pregnancy rates in primiparous dairy cows. The aim of the current study was to confirm the success of the Doublesynch protocol and further to investigate the effect of this method on pregnancy rates in anestrous cows. Lactating primiparous Holstein (Bos taurus) cows (n = 165) between 60 and 172 d postpartum were monitored twice with 10-d intervals (on Days -10 and 0) by ultrasonography, and blood samples were collected. Cows were classified as anestrous if both blood samples had progesterone (P4) concentration <1 ng/mL and as cyclic if at least one of the two samples had P4 concentration ≥1 ng/mL. Cyclic cows were classified again as cyclic-high P4 (having an active corpus luteum) if the second blood samples had P4 concentrations ≥1 ng/mL and as cyclic-low P4 if P4 concentrations were <1 ng/mL on Day 0. Then, the cows classified as anestrous (n = 51), cyclic-high P4 (n = 63), or cyclic-low P4 (n = 51) were put into two treatment groups (Ovsynch or Doublesynch) randomly to establish six groups. Cows in the Ovsynch group were administered a GnRH (lecirelin 50 μg, im) on Day 0, PGF (Prostaglandin F2 alpha, D-cloprostenol 0.150 mg, im) on Day 7, and a second dose of GnRH 48 h later. Cows in the Doublesynch group were administered a PGF on Day 0, GnRH on Day 2, a second PGF on Day 9, and a second GnRH on Day 11. Timed artificial insemination (TAI) was performed 16 to 20 h after the second GnRH in both treatment groups. Pregnancy diagnosis was conducted (by ultrasonography) 45 ± 5 d after TAI. In anestrous cows and those with high and low progesterone concentration at treatment onset, Doublesynch treatment led to markedly increased pregnancy rates with respect to Ovsynch treatment (P < 0.05). On the overall analysis of data, it was revealed that the Doublesynch method increased pregnancy rates by 43 percentage units (29.8% vs. 72.8%, P < 0.0001) in relation to Ovsynch. Pregnancy rates of cows having small, medium, or large follicles at the day of second GnRH administration were similar in the Doublesynch group (70.4%, 85.2%, and 63.0%, respectively; P > 0.05), whereas pregnancy rates reduced dramatically as follicle size increased in the Ovsynch group, particularly in cows with follicles greater than 16 mm (45.5%, 28.1%, and 5.3%, respectively; P < 0.05). Our results confirm and support observations that the Doublesynch protocol increases the pregnancy rates in postpartum primiparous cows as reported previously. Our data also demonstrate that the Doublesynch method increases the pregnancy rates in anestrous cows. Thus, these data suggest that the Doublesynch protocol can be used to obtain satisfactory pregnancy rates after TAI in both anestrous and cycling primiparous dairy cows regardless of stage of estrous cycle.     

Persistent ovarian follicles in dairy cows: a therapeutic approach

Anestrus is common during the postpartum period in high-producing dairy cows. In a previous investigation, we were able to diagnose persistent follicles of 8 to 12 mm in anestrous cows. This report describes 2 consecutive studies. The objectives of the first were to 1) assess the association of persistent follicles with anestrus; and 2) evaluate 2 therapeutic treatments. In the second study, we compared the effectiveness of the best treatment established in Study 1 with the Ovsynch protocol. For Study 1, anestrous cows were considered to have a persistent follicle if it was possible to observe a single follicular structure > 8 mm in the absence of a corpus luteum or a cyst in 2 ultrasonographic examinations performed at an interval of 7 d. At diagnosis (Day 0), cows were assigned to 1 of 3 treatment groups. Cows in Group GnRH/PGF (n=17) were treated with 100 microg GnRH i.m., and 25 mg PGF2alpha i.m. on Day 14. Cows in Group PRID (n=18) were fitted with a progesterone releasing intravaginal device (PRID, containing 1.55 g of progesterone) for 9 d and were given 100 microg GnRH i.m. at the time of PRID insertion, and 25 mg PGF2alpha i.m. on Day 7. Cows in Group Control (n=18) received no treatment. The animals were inseminated at observed estrus and were monitored weekly by ultrasonography until AI or 5 weeks from diagnosis. Blood samples were also collected on a weekly basis for progesterone determination. The mean size of persistent follicles on Day 0 was 9.4 +/- 0.04 mm. Progesterone levels were < 0.2 ng/mL during the first 35 d in 16 of 18 Control cows. Cows in the PRID group showed a lower persistent follicle rate (16.7% < 70.6% < 88.9%; P < 0.0001; PRID vs GnRH/PGF vs Control, respectively); a higher estrus detection rate (83.3% > 29.4% > 11.1%; P < 0.0001) and a higher pregnancy rate (27.8% > 5.9% > 0%; P = 0.02). For the second study, 145 cows with persistent follicles were randomly assigned to 1 of 2 treatment groups: cows in Group Ovsynch (n=73) were treated with 100 microg GnRH i.m. on Day 0, 25 mg PGF2alpha i.m. on Day 7, and 100 microm GnRH i.m. 32 h later. Cows in this group were inseminated 16 to 20 h after the second GnRH dose (Ovsynch protocol). Cows in Group PRID (n=72) were treated as those in the PRID group of Study 1, and were inseminated 56 h after PRID removal. Cows in the PRID group showed a higher ovulation rate (84.8% > 8.2%: P < 0.0001); a higher pregnancy rate (34.2% > 4.1%; P < 0.0001) and lower follicular persistence rate (22.2% < 63%; P < 0.0001) than those in Ovsynch. Our results indicate that persistent follicles affect cyclic ovarian function in lactating dairy cows. Cows with persistent follicles can be successfully synchronized and time inseminated using progesterone, GnRH and PGF2alpha but show a limited response to treatment with GnRH plus PGF2alpha.     

A progesterone-based timed AI protocol more effectively prevents premature estrus and incomplete luteal regression than an Ovsynch protocol in lactating Holstein cows

The objective of this study was to evaluate pregnancy rates in lactating Holstein cows treated with an Ovsynch protocol (GnRH-PGF(2alpha)-GnRH) or a progesterone-based timed AI (TAI) protocol, and to determine the factors that may influence pregnancy rate following protocol treatment. In experiment 1, lactating Holstein cows were randomly assigned to three treatments: (1) an injection of GnRH (Day 0), an injection of PGF(2alpha) on Day 7, a second injection of GnRH on Day 9, and TAI 16h after the second GnRH injection (GPG group, n = 34); (2) insertion of a CIDR intravaginal progesterone (1.9g) device combined with a capsule containing 10mg estradiol benzoate (Day 0), an injection of PGF(2alpha) and removal of the device on Day 7, an injection of GnRH on Day 9, and TAI 16h after the GnRH injection (CPG group, n = 34); (3) an injection of PGF(2alpha) after confirming the presence of CL by ultrasonographical observation and artificial insemination at estrus (AIE) (P group, n = 75). The pregnancy rate after TAI following the CPG protocol (41.2%) was higher (P<0.05) than that after TAI following the GPG protocol (20.6%) and that after AIE (20.0%). In experiment 2, lactating Holstein cows were randomly assigned to two treatments: a GPG group (n = 31) and a CPG group (n = 31). The GPG and CPG protocols were identical to those used in experiment 1. The proportion of cows with premature estrus prior to injection of PGF(2alpha) and with incomplete luteal regression tended (P = 0.056) to be or were greater (P<0.05) in the GPG group (4/31, 8/31) than in the CPG group (0/31, 2/31), respectively. Average diameters of dominant follicles (1.5+/-0.1mm versus 1.4+/-0.1mm) on Day 7 and preovulatory follicles (1.8+/-0.1mm versus 1.6+/-0.1mm) on Day 9, and the proportion of cows with synchronized ovulation by 40h after the second GnRH injection were not different (81.5% versus 87.1%, P>0.05) between groups, respectively. We conclude that the pregnancy rate after TAI following the CPG protocol was higher than that after TAI following the GPG protocol, probably due to a decreased incidence of premature estrus and incomplete luteal regression.     

Fertility of swamp buffalo following the synchronization of ovulation by the sequential administration of GnRH and PGF2alpha combined with fixed-timed artificial insemination

This study evaluated fertility in swamp buffalo after synchronization of ovulation combined with fixed time artificial insemination. At the start of the study, designated day 0, from a group of 98 female Thai swamp buffalo, 55 buffalo (heifers n° = 20 and cows n° = 35) were selected to be synchronized with GnRH (Day 0) followed by PGF₂alpha (Day 7) and a second treatment with GnRH (Day 9). All buffalo were inseminated at two fixed times 12 h and 24 h after the second injection of GnRH (Ovsynch+TAI group); a second group of 43 buffalo (heifers n° = 19 and cows n° = 24) were not treated and were artificially inseminated (AI) at natural estrus (AI group). Blood samples were taken 22 days after insemination to evaluate progesterone plasma levels. In the Ovsynch+TAI group, overall conception rate (CR; i.e. the number of cows with progesterone >4.0 ng/ml on day 22 after AI divided by the number of animals inseminated), was 38.1% and overall pregnancy rate (PR; i.e. the number of cows that were pregnant at day 50-60 after insemination divided by the number of animals inseminated), was 32.7%. In the AI group overall CR and PR was 34.9%.Within the Ovsynch+TAI group, CR and PR were reduced (P < 0.05) in heifers compared with cows (CR 15.0% vs. 51.4% for heifers and cows, respectively; PR 15.0% vs. 42.9% for heifers and cows, respectively). Within the AI group the efficacy of treatment was similar between heifers and cows (CR and PR 31.6% for heifers and 37.5% for cows).In conclusion, this study indicates that in swamp buffalo it is possible to synchronize ovulation and use timed artificial insemination with the Ovsynch+TAI protocol.     

Reproductive performance of dairy cows with ovarian cysts after different GnRH and cloprostenol treatments

Cystic ovarian disease is an important cause of reproductive failure and economic loss for the dairy industry. This report describes two consecutive studies. The objective of the first was to evaluate the response of cows with ovarian cysts to two therapeutic treatments. In the second study, we compared the effectiveness of the best treatment established in Study 1 with that of the Ovsynch protocol. For Study 1, cows were considered to have an ovarian cyst if it was possible to observe a single follicular structure with a follicular antrum diameter > 25 min in the absence of a corpus luteum in three ultrasonographic examinations performed at 7 days intervals. At diagnosis (Day 0), cows were assigned to one of two treatment groups. Cows in Group GnRH/CLP (n = 31) were treated with 100 microg GnRH i.m. and 500 microg cloprostenol (CLP) i.m. on Day 14. Cows in Group GnRH-CLP/CLP(n = 32) were treated with 100 microg GnRH i.m. plus 500 microg CLP i.m. on Day 0, and 500 microg CLP i.m. on Day 14. The animals were inseminated at observed estrus and monitored weekly by ultrasonography for 4 weeks or until Al. Cows in the GnRH-CLP/CLP group showed a lower cystic persistence rate (15.6% < 45.2%; P = 0.01); a higher estrus detection rate (84.4% > 41.9%; P < 0.0001); a higher ovulation rate (75% versus 32.3%; P < 0.0001) and a higher early response rate (31% > 3%; P = 0.02) than those in the GnRH/CLP group. For the second study, 128 cows with ovarian cysts were randomly assigned to one of two treatment groups: cows in Group Ovsynch (n = 64) were treated with 100 microg GnRH i.m. on Day 0, 500 microg CLP on Day 7, and 100 microm GnRH i.m. 36 h later. Cows in this group were inseminated 24 h after the second GnRH dose (Ovsynch protocol). Cows in Group GnRH-CLP/CLP/GnRH (n = 64)were treated as those in the GnRH-CLP/CLP group of Study 1 but received GnRH 32 h after the second CLP treatment and were inseminated 24 h after this. A further group of cows without ovarian cysts inseminated at natural estrus served as the Group Control (n = 64). Cows in the GnRH-CLP/CLP/ GnRH group showed a lower cystic persistence rate (10.9% < 46.9%; P < 0.0001); higher ovulation rate (79.7% > 17.2%; P < 0.0001); higher return to estrus rate (34.3% > 12.5%; P < 0.01) and higher pregnancy rate (28.1% > 3.1%; P < 0.01) than those in Ovsynch; and a similar pregnancy rate (28.1% versus 35.9%) to Control cows. These findings indicate that lactating cows with ovarian cysts can be successfully synchronized and time inseminated using a protocol that combines GnRH and CLP, starting treatment by simultaneously administering both products. This protocol also allows the insemination of cows showing estrus within the first week of treatment. Ovarian cysts were less responsive when treatment was started with GnRH alone.     

Modifications of the G6G timed-AI protocol improved pregnancy per AI and reduced pregnancy loss in lactating dairy cows

In dairy cows, subjected to a G6G protocol, objectives were to determine effects of (1) extending the interval from prostaglandin F2α (PGF2α) to gonadotropin-releasing hormone (GnRH) during presynchronization; and (2) adding a second PGF2α treatment before artificial insemination (AI), on ovarian response, plasma progesterone (P4) concentrations and pregnancy per AI (P/AI). In a 2×2 factorial design, lactating cows were randomly assigned to one of four timed AI (TAI) protocols: (1) G6G (n=149), one injection of PGF2α, GnRH 2 days later and a 7-day Ovsynch (GnRH, 7 days, PGF2α, 56 h, GnRH, 16 h, TAI) was initiated 6 days later; (2) G6GP (n=144), an additional PGF2α treatment (24 h after the first) during Ovsynch of the G6G protocol; (3) MG6G, one injection of PGF2α, GnRH 4 days later before initiation of the G6G protocol; and (4) MG6GP, an additional PGF2α treatment (24 h after the first) during Ovsynch of the MG6G protocol. Blood samples were collected (subset of 200 cows) at first GnRH and PGF2α of the Ovsynch, and at TAI to measure P4. Ultrasound examinations were performed in a subset of 406 cows to evaluate ovarian response at various times of Ovsynch, and in all cattle to determine pregnancy status at 32 and 60 days after TAI. Extending the interval by 2 days between PGF2α and GnRH during presynchronization increased (P<0.01) ovulatory response to first GnRH of Ovsynch, circulating P4 during Ovsynch, and P/AI at 32 and 60 days after TAI. Adding a second PGF2α treatment before AI increased the proportion of cows with luteal regression (P=0.04), improved P/AI at 60 days after TAI (P=0.05), and reduced pregnancy loss between 30 and 60 days after TAI (P=0.04). In summary, extending the interval from PGF2α to GnRH during presynchronization increased response to first GnRH of Ovsynch and P4 concentrations during Ovsynch, whereas adding a second PGF2α treatment before AI enhanced luteal regression. Both modifications of the G6G protocol improved fertility in lactating dairy cows.     

Strategies to optimize reproductive efficiency by regulation of ovarian function in yak (Poephagus grunniens L.)

Ovulatory response to the first GnRH of Ovsynch is the critical determinant for successful synchronization of ovulation in animals. An attempt was made in this study to design a pre-Ovsynch hormonal strategy in yaks to increase the ovulatory response to the first GnRH injection of Ovsynch so that overall synchronization rate to Ovsynch could be improved. Non-lactating cyclic yak cows (n=33) were assigned to receive either no treatment before Ovsynch (control) or 0.375 mg of PGF2alpha (PreP) followed 2 d later by 10 microg of GnRH (PreG), administered 4 (G4G), 5 (G5G), or 6 (G6G) d before initiating the Ovsynch protocol. Rectal palpation was performed to assess ovulation and blood samples were collected to measure progesterone concentrations during pre-treatment, treatment and post-treatment periods. All the animals received timed AI 12 and 24h after the final GnRH of Ovsynch. Diagnoses for pregnancy were performed by rectal palpation and profiles of plasma progesterone 35 d after AI. Percentage of yak cows that ovulated in response to the first GnRH injection of Ovsynch, synchronized to Ovsynch treatment, had a functional CL at PGF2alpha of Ovsynch, had circulating concentrations of P4 at PGF2alpha of Ovsynch and likely to be pregnant after 35 d after AI, were greater in G6G and G5G compared with control, whereas G4G did not differ from controls. In addition, animals that ovulated in response to first GnRH of Ovsynch had greater response to PGF2alpha of Ovsynch and greater synchronization rate to the overall protocol than those that did not ovulate. In summary, PGF2alpha-and-GnRH-based pre-Ovsynch strategies consisting of 5 or 6-d interval between PreG and first GnRH of Ovsynch resulted in a greater ovulatory and luteolytic response to first GnRH andPGF2alpha of Ovsynch, respectively, compared with control animals. These, in turn, optimized synchronization rate to Ovsynch in yaks.     

Effect of a deslorelin implant in a timed artificial insemination protocol on follicle development, luteal function and reproductive performance of lactating dairy cows

This study examined the influence of a GnRH agonist containing either 450 or 750 microg of deslorelin in an implant form or a gonadorelin injection (control) to induce ovulation in the Ovsynch protocol on pregnancy rates (PR), embryonic loss, and ovarian function in 593 lactating Holstein cows. Cows were given two injections of PGF2alpha 14 days apart, followed 14 days later by the Ovsynch protocol, and were timed artificially inseminated (TAI) at 68 +/- 3 days postpartum. Blood samples for determination of plasma progesterone concentrations were collected at 24 and 10 days prior to and 11 days after TAI. Pregnancy was diagnosed on Day 27 and reconfirmed on Day 41 after TAI. Non-pregnant, not re-inseminated cows at Day 27 had their ovaries examined by ultrasonography, and the number and size of follicles and presence of luteal tissue were determined. Simultaneously, these cows were re-synchronized with the Ovsynch protocol. Pregnancy during the re-synchronization period was determined between 35 and 41 days after insemination. On Day 27, PR were higher for control (39.0%) and deslorelin 450 microg (DESLORELIN 450) implant (41.3%) than for those receiving the deslorelin 750 microg (DESLORELIN 750) implant (27.5%; P<0.05). Pregnancy losses tended to decrease for DESLORELIN 450 compared with control (5.0% versus 12.7%; P<0.13). Plasma progesterone concentrations did not differ significantly among treatments. Deslorelin suppressed ovarian activity and decreased PR during the re-synchronization period compared with control. The percentage of non-pregnant animals that were re-inseminated by Day 27 was less for deslorelin compared with control. In conclusion, incorporation of an implant of the GnRH agonist deslorelin to induce ovulation in the Ovsynch protocol has the potential to reduce pregnancy losses, but the response was dependent upon implant concentration. Evaluation of lower doses to minimize the negative effects on subsequent fertility is warranted.     

Ovarian follicular wave synchronization and induction of ovulation in postpartum beef cows

An experiment was designed to evaluate a) the effect of a progesterone-estradiol combined treatment on ovarian follicular dynamics in postpartum beef cows, and b) ovulation and the subsequent luteal activity after short-term calf removal and GnRH agonist treatment. Multiparous Angus cows (25 to 40 d after calving) were assigned to the following treatments: untreated (Control, n = 9); short term calf removal (CR, n = 8); progesterone (CIDR, n = 9) and progesterone plus estradiol-17 beta (CIDR + E-17 beta, n = 9). Progesterone treatment (CIDR) lasted 8 d and the day of device insertion was considered as Day 0. Cows in the CIDR + E-17 beta group also received an i.m. injection of 5 mg of E-17 beta on Day 1. On Day 8, calves were removed for 48 h (CR, CIDR and CIDR + E-17 beta groups) and 6 h before the end of calf removal these cows also received an i.m. injection of 8 micrograms of Busereline (GnRH). Anestrus was confirmed in all cows by the absence of luteal tissue and progesterone concentrations below 1 ng ml-1 at the beginning of the experiment. Although mean (+/- SEM) interval from the beginning of the experiment (Day 0) to wave emergence did not differ (P > 0.05) among treatment groups (Control, 1.9 +/- 1.0, range -2 to 7 d; CR, 3.9 +/- 0.7, range 0 to 6 d; CIDR, 2.8 +/- 0.5, range 0 to 4 d and CIDR + E-17 beta, 4.1 +/- 0.2, range 3 to 5), the variability was less (P < 0.05) in the CIDR + E-17 beta group. The proportion of cows ovulating 24 to 48 h after GnRH administration tended (P = 0.08) to be higher in cows from CIDR + E-17 beta group (8/9) than in those of CR (5/8) or CIDR (6/9) groups, respectively and was associated with a higher proportion (P < 0.05) of CIDR + E-17 beta treated cows (9/9) that had a dominant follicle in the growing/early static phase at the time of GnRH treatment compared to the other GnRH treated groups (5/8, and 4/9 for CR and CIDR groups, respectively). Two CR cows ovulated 0-24 h after GnRH and only one Control cow ovulated the day before the time of GnRH administration. Cows pretreated with progesterone had longer (P < 0.05) luteal lifespan (CIDR, 14.5 +/- 0.7, CIDR + E-17 beta, 13.9 +/- 0.6 d) than those not treated with CIDR (Control, 5, CR, 4.0 +/- 0.4). We conclude that progesterone plus estradiol treatment results in tightly synchronized wave emergence and high GnRH-induced ovulation rate with normal luteal activity in postpartum beef cattle.     

The effect of aspirin administration and parity on plasma salicylate concentrations and postpartum reproductive parameters in Brahman cows

Forty pluriparous (M) and 20 primiparous (P) suckled Brahman cows were used to evaluate the effect of aspirin and parity on plasma 13,14-dihydro-15-keto-prostaglandin F2alpha (PGFM) and progesterone (P4) concentrations and some reproductive parameters. On Day 7 after calving (PP), the cows were allocated within parity into 2 groups: the aspirin group received concentrate containing aspirin at a rate of 100 mg/kg of body weight every 12 h until Day 13 PP; and the control received concentrate every 12 h for the same interval. Blood samples were collected after first and last aspirin feeding and daily from Day 1 PP to Day 6 PP and from Day 14 PP to Day 21 PP, twice daily from Day 7 PP to Day 13 PP, and weekly until first non-return to estrus. Plasma salicylate concentrations in the aspirin group cows were affected by parity (P < 0.01) and time after feeding (P < 0.0001). P cows showed higher plasma salicylate concentrations with a later peak and slower decrease than M cows. Aspirin-treated P cows had longer PP intervals than either control P, control M, or aspirin-treated M cows. Cows receiving aspirin had a lower pregnancy rate, an increased incidence of abnormal estrous cycles, and a decline in the presence of corpora lutea after estrus. Cows that formed a corpora lutea and had received aspirin had higher P4 release between Day 6 and 14 after estrus. Aspirin-treated cows that did not form corpora lutea had lower P4 release between Days 9 and 14 after estrus. A treatment by parity interaction affected mean PGFM proportions (P < 0.01) during the treatment period. Aspirin-fed P cows increased PGFM release as measured by mean proportion of Day 6 PP values. Aspirin-fed M cows showed a decrease in mean PGFM proportions. Aspirin feeding during the early PP showed different effects on some reproductive parameters in P and M Brahman cows, indicating differences in PP physiology between parities.     

Ultrasonographic evaluation of endometrial thickness near timed AI as a predictor of fertility in high-producing dairy cows

The objectives were to evaluate changes in endometrial thickness (ET) near the time of a synchronized ovulation and to assess the relationship of ET and fertility in lactating Holstein cows, with or without estrogen supplementation near timed ovulation. In Experiment 1, eight cows were examined with transrectal ultrasonography, once daily for 5 d, starting concurrent with PGF_2α (PGF) treatment during an Ovsynch protocol (GnRH - 7d - PGF - 72h - GnRH). The ET increased rapidly after PGF (from ∼7 to ∼9.5 mm), remained > 9 mm for the next 2 d, then decreased to ∼8 and 7.4 mm, 1 and 2 d, respectively, after the second GnRH. In Experiment 2,642 cows (total of 758 breedings) were subjected to an Ovsynch protocol (GnRH - 7d - PGF - 56h - GnRH - 16h - timed AI); cows received either no further treatment (Ovsynch) or 1 mg of estradiol-17β im 8 h before the second GnRH (Ovsynch + E2). For both uterine horns, ET was measured (∼2 cm from the internal uterine body bifurcation) before E2 treatment (48 h after PGF). In cows with ET ≤ 8 mm vs > 8 mm, rates of ovulation were 86.0% (n = 136) vs 98.1% (n = 472; P < 0.01), respectively, and percentage pregnant per AI (P/AI) were 26.7% (n = 146) vs 42.7% (n = 524; P < 0.01). Treatment with E2 increased P/AI in cows with lower ET (Ovsynch + E2 = 37.0% vs Ovsynch = 23.3%; P = 0.07), but did not significantly improve P/AI in cows with ET > 8 mm (Ovsynch + E2 = 43.4% vs Ovsynch = 42.1%). In conclusion, a single ultrasonographic evaluation of ET in Holstein cows 48 h after PGF treatment in an Ovsynch program was a good predictor of ovulation failure and pregnancy success. Perhaps poor fertility in cows with reduced ET was low peripheral E2 concentrations near AI, poor P4 priming, or luteolysis failure during timed AI procedures.     

Effect of pretreatment with bovine somatotropin (bST) and/or gonadotropin-releasing hormone (GnRH) on conception rate of dairy cows with ovarian cysts subjected to synchronization of ovulation and timed insemination

The objective of this study was to determine the effect of pretreatment with bovine somatotropin (bST) and/or gonadotropin-releasing hormone (GnRH) 7 days prior to initiation of a protocol for synchronization of ovulation and timed insemination (Ovsynch) on conception rate (CR) of cows with ovarian cysts. A total of 254 lactating dairy cows with ovarian cysts was divided into four groups (Day 0). On Day 0, cows in Group 1 (n = 61) were pretreated with 500 mg bST, s.q., and 100 microg GnRH, i.m.; cows in Group 2 (n = 73) were pretreated with 100 microg GnRH, i.m.; cows in Group 3 (n = 59) were pretreated with 500 mg bST, s.q.; and cows in Group 4 (n = 61) received no pretreatment. All cows were subjected to the Ovsynch protocol 7 days later. All cows previously received routine bST treatment every 14 days until milk production decreased to a minimum level established by the management of the herd. CR was assessed using logistic regression after adjusting for timing of concurrent bST treatment relative to Day 0, parity, season at time of insemination, and days in milk (DIM) on Day 0. CR for cows in Group 3 (12%) was significantly lower (P < 0.05) than that for cows in Group 4 (27%), and CR for cows in Group 1 (18%) and Group 2 (15%) tended to be lower (P < 0.10) than that for cows in Group 4 (27%). From the results of this study, it was concluded that bST pretreatment decreased CR, and pretreatment with GnRH, and GnRH with bST tended to decrease CR in lactating dairy cows with ovarian cysts concurrently treated with bST and subjected to the Ovsynch protocol.     

Ovsynch synchronization and fixed-time insemination in goats

This study assessed the efficacy of an Ovsynch protocol (vs. the classical cronolone containing vaginal sponge+eCG treatment) to generate fixed-time insemination in goats during the breeding season. Each regimen was applied to 24 Boer goat does. Onset and duration of estrus were determined with an aproned male and follicular development was monitored by ultrasonography. Ovulation and quality of the corpora lutea were established from progesterone concentrations. In 10-11 goats per group, LH concentrations were determined throughout the preovulatory period. Does were inseminated at pre-determined times (16 h after the second GnRH injection and 43 h after sponge removal). Estrus was identified in 96% of the Ovsynch-treated goats (at 49 h after prostaglandin injection) and in 100% of the goats synchronized with sponges (at 37 h after sponge removal). Low progesterone concentrations at the time of AI were observed in 21/24 and 24/24 goats synchronized by Ovsynch and sponges, respectively. Synchronization of the LH surge was tighter following Ovsynch compared to sponge treatment. Kidding rates (at 58 and 46% in the Ovsynch and sponge groups, respectively) and prolificacy (at 1.86 and 1.83 in the Ovsynch- and sponge-treated goats) were similar for both groups, as were the number of ovulations (2.9 and 3.3) and the proportion of does with premature corpus luteum regression (29 and 17%). When excluding does with premature luteal regression and those with low progesterone levels when receiving prostaglandins, kidding rate reached 87.5% (14/16) after Ovsynch. During the breeding season, the Ovsynch protocol may thus be an useful alternative to the sponge-eCG treatment.