Progesterone-releasing Intravaginal Device Research Articles

CASE REPORT: A SUCCESSFUL TREATMENT STRATEGY FOR CYSTIC OVARIAN DISEASE IN A DAIRY COW

A 5-year-old multiparous lactating Holstein cow was reported to have cystic ovarian disease with a history of whitish vaginal discharge on the 80th day postpartum. On the 83rd day postpartum, transrectal ultrasonography revealed a thick-walled cystic structure with visible cobwebs in the cavity, measuring 4.2[Formula: see text]cm × 3.4[Formula: see text]cm in diameter on the right ovary. The serum progesterone (P4) concentration was 6.82[Formula: see text]ng/mL, leading to a diagnosis of luteinized follicular cyst. Transvaginal aspiration of the cyst was performed to decrease the compression of ovarian stroma by the cyst, followed by intramuscular administrations of cloprostenol and buserelin. A 1.9[Formula: see text]cm diameter corpus luteum was detected on the right ovary nine days after aspiration, with a serum P4 concentration of 1.29[Formula: see text]ng/mL. Continual hormone treatment was followed, consisting of an injection of buserelin and an intravaginal progesterone-releasing device inserted for seven days. After seven days, two corpora lutea were noted on the right ovary with an average diameter of 2.8[Formula: see text]cm and 1.8[Formula: see text]cm, respectively. The device was removed and the cow showed a P4 concentration of 11.22[Formula: see text]ng/mL on that day. Estrus induction was conducted with two low doses of cloprostenol followed by an injection of gonadorelin, to induce a complete luteolysis and the following ovulation to fit a fixed-timed artificial insemination (AI). Pregnancy was confirmed 33 days after AI. Although the number of days open was 102 in this case, it took a total of 19 days from treatment to pregnancy.

Comparison of Gonadotropin-Releasing Hormone versus Estrogen-Based Fixed-Time Artificial Insemination Protocols in Grazing Bos taurus Suckled Beef Cows.

Fixed-timed artificial insemination (FTAI) protocols for beef cattle in South America are primarily based on estradiol esters and intravaginal progesterone-releasing devices (IVPD). The objective of this study was to determine the optimal gonadotropin-releasing hormone (GnRH)-based protocol as an alternative to the use of estrogen-based protocols in grazing Bos taurus suckling beef cows. All cows received an IVPD on the day of protocol initiation and prostaglandin F2α (PG) plus equine chorionic gonadotropin (eCG) treatments at the time of IVPD removal. In Experiment 1, cows (n = 235) were randomly assigned to one of four treatments: (i) 7-day estradiol = 2 mg of estradiol benzoate (EB) at IVPD insertion on Day 9 and 1 mg of estradiol cypionate (ECP) at IVPD removal on Day 2; (ii) 7-day GnRH = 10 µg of GnRH at IVPD insertion on Day 10, IVPD removal on Day 3 and GnRH at FTAI; (iii) 7 & 7 estradiol = PG at IVPD insertion on Day 16, EB on Day 9 and ECP at IVPD removal on Day 2; (iv) 7 & 7 GnRH = PG at IVPD insertion on Day 17, GnRH on Day 10, IVPD removal on Day 3 and GnRH at FTAI. In Experiment 2, cows (n = 462) were randomly assigned to one of four treatments: (i) 6-day estradiol = EB at IVPD insertion on Day 9, IVPD removal on Day 3 and GnRH at FTAI; (ii) 7-day estradiol; (iii) 7-day GnRH; (iv) 7 & 7 GnRH. In Experiment 1, plasma progesterone concentrations and percentage of cows with a corpus luteum (CL) at IVPD removal, and pregnancy per AI (P/AI) were greater for cows subjected to GnRH-based protocols compared with cows subjected to estrogen-based protocols (p < 0.01). In Experiment 2, cows subjected to the 7 & 7 GnRH protocol had the greatest P/AI (p < 0.01). In summary, GnRH-based FTAI protocols resulted in similar or greater P/AI compared to estrogen-based FTAI protocols in grazing postpartum Bos taurus suckled beef cows. The greatest P/AI was attained with the 7 & 7 GnRH protocol.

Evaluation of minimally invasive estrus synchronization protocols in brown brocket deer (Subulo gouazoubira)

This study aimed to evaluate minimally invasive protocols for estrus synchronization in the brown brocket deer (Subulo gouazoubira). Females were submitted to Latin square design, in different treatments. All females received 0.25 mg of estradiol benzoate on the first day of treatment, concomitant with one of the following sources of progesterone: (1) DIP: an intravaginal progesterone releasing device for eight days, (2) MGA1x: once a day (in the morning) oral dose of 1 mg melengestrol acetate for eight days, (3) MGA2x: twice a day (morning and afternoon) oral doses of 0.5 mg of MGA for eight days, (4) P4LA: a single i.m. administration of 75 mg of long-acting progesterone (P4LA). Eight days after the beginning of each treatment, females received an i.m. administration of 265 µg of prostaglandin (PGF2α; cloprostenol). Treatment efficacy was evaluated by manifestation of behavioral estrus after treatment and concentration of fecal progesterone metabolites (FPM). The time to onset of estrus in treatment P4LA was significantly longer (180 ± 38.9 h) compared to DIP (63 ± 6.6 h), MGA1x (53 ± 14.4 h) and MGA2x (41 ± 10.1 h) (P = 0.008). According to individual baseline FPM and FPM concentration during the days after estrus, the corpus luteum formation was suggested in all females which responded to the treatments (93.75 %). Low synchrony, longer interval between PGF2α administration and onset of estrus suggest that the P4LA dose (75 mg) is too high and not effective for S. gouazoubira. DIP, MGA 1x and MGA 2x, were effective in estrus synchronization.

Effect of delayed timing of artificial insemination with sex-sorted semen on pregnancy per artificial insemination in synchronized dairy heifers managed in a seasonal-calving pasture-based system

The objective of this study was to evaluate the timing of artificial insemination (AI) with frozen-thawed sex-sorted semen on pregnancy per AI (P/AI) in dairy heifers. A 6-d progesterone Co-Synch protocol was used for ovulation synchronization of dairy heifers, with timed AI (TAI) coincident with (TAI-0) or 8 h (TAI-8) after the second injection of GnRH, corresponding to either 48 h or 56 h after removal of the progesterone-releasing intravaginal device. Pregnancy diagnosis was conducted by transrectal ultrasound scanning of the uterus 34 d after TAI (n = 816 records available for analysis). Generalized linear mixed models were used to examine the effects of treatment on P/AI. Treatment (n = 2), herd (n = 11), and treatment × herd were included as categorical fixed effects. Heifer body weight and Economic Breeding Index values for milk production, fertility, calving performance, beef carcass, cow maintenance, cow management, and health were included as continuous fixed effects. Heifer ID was included as a random effect. Pregnancy per AI was greater for TAI-8 heifers (59%) compared with TAI-0 heifers (50%). Pregnancy per AI ranged from 38% to 75% between herds but there was no treatment × herd interaction. The fertility subindex (positive) and the cow management subindex (negative) were the only continuous animal variables associated with P/AI. Delaying the timing of AI with frozen-thawed sex-sorted semen by 8 h in dairy heifers enrolled on a 6-d progesterone Co-Synch protocol improved P/AI.

Estrous activity and pregnancy outcomes in Holstein heifers subjected to a progesterone based 5-d CO-Synch protocol with or without administration of initial GnRH.

The objectives of this study were to investigate whether estrous activity and its impact on pregnancy outcomes would differ in heifers subjected to a 5d CO-Synch plus progesterone releasing intravaginal device (PRID) protocol with or without an initial GnRH treatment. Holstein Heifers (n=308) were fitted with a collar-mounted automated activity monitoring system approximately 1 week prior to the initiation of the synchronization protocol (Day-7). Heifers were assigned randomly to a 5d CO-Synch plus PRID protocol either with (GnRH; n=154) or without (NGnRH; n=154) an initial administration of 100μg of GnRH at the time of PRID insertion (Day 0). Heifers received a single administration of 500μg of cloprostenol (PGF) at the time of PRID removal (Day 5) and again 24h later (Day 6). Approximately 72h after PRID removal (Day 8), heifers were timed-inseminated (TAI) and concurrently 100μg of GnRH was administered to those not exhibiting estrus. All inseminations were done by one of two technicians using either sex-sorted (n=252) or conventional (n=56) frozen-thawed semen. Transrectal ultrasonography was done on Day 0 to determine ovarian cyclicity and normalcy of the reproductive tract and 30 and 45d post-TAI to determine and confirm pregnancy, respectively. The percentage of heifers determined to be in estrus following PRID removal was greater in the GnRH than in the NGnRH group (94 vs. 82%, respectively; P<0.01). The mean interval from PRID removal to the onset of estrus was shorter for GnRH- than NGnRH-treated heifers (50.8 vs. 59.2h, respectively; P<0.01). Pregnancy per AI (P/AI) at 30d post-TAI tended to be greater for GnRH than NGnRH heifers (68 vs. 59%, respectively; P=0.1). However, P/AI at 45d post-TAI (65 vs. 57%, respectively) and pregnancy loss between 30 and 45d post-TAI (6 vs. 4.5%, respectively) did not differ. The association between the interval from PRID removal to the onset of estrus and P/AI at 30d post-TAI was linearly negative for GnRH heifers; for every 1h increase in the interval from PRID removal to the onset of estrus, the predicted probability of P/AI at 30d post-TAI tended (P=0.08) to be reduced by 2.7%. The association between the interval from PRID removal to the onset of estrus and P/AI at 30d post-TAI was not significant for NGnRH heifers. In addition, the interval from TAI to subsequent estrus, in non-pregnant heifers, was approximately 3d longer for the GnRH than the NGnRH group (20.7 vs. 17.5d, respectively). In summary, the initial GnRH treatment in a 5d CO-Synch plus PRID protocol increased estrus expression and reduced the interval from PRID removal to the onset of estrus in Holstein heifers and tended to increase P/AI at 30d, but did not affect P/AI at 45d post-TAI.

Impact of oestrus synchronization devices on ewes vaginal microbiota and artificial insemination outcome.

The low pregnancy rate by artificial insemination in sheep represents a fundamental challenge for breeding programs. In this species, oestrus synchronization is carried out by manipulating hormonal regimens through the insertion of progestogen intravaginal devices. This reproductive strategy may alter the vaginal microbiota affecting the artificial insemination outcome. In this study, we analyzed the vaginal microbiome of 94 vaginal swabs collected from 47 ewes with alternative treatments applied to the progesterone-releasing intravaginal devices (probiotic, maltodextrin, antibiotic and control), in two sample periods (before placing and after removing the devices). To our knowledge, this is the first study using nanopore-based metagenome sequencing for vaginal microbiome characterization in livestock. Our results revealed a significant lower abundance of the genera Oenococcus (Firmicutes) and Neisseria (Proteobacteria) in pregnant compared to non-pregnant ewes. We also detected a significant lower abundance of Campylobacter in the group of samples treated with the probiotic. Although the use of probiotics represents a promising practice to improve insemination results, the election of the suitable species and concentration requires further investigation. In addition, the use of progestogen in the synchronization devices seemed to increase the alpha-diversity and decrease the abundance of harmful microorganisms belonging to Gammaproteobacteria and Fusobacteriia classes, suggesting a beneficial effect of their use.

Ovine endometrial estrogen receptor expression is altered following PG-600 administration.

Dysregulation of the estrogen receptor 1 (ESR1) expression during the establishment of pregnancy could contribute to reduce fertility reported in ewes treated with PG-600. The objective of this study was to evaluate the effect of treatment with PG-600 on ESR1 expression in the ovine endometrium during early diestrus. Polypay ewes (n = 24) that had exhibited an oestrous cycle of normal duration (16-18 days) were treated with intravaginal progesterone-releasing device (CIDR) for 9 days. Cloprostenol was administrated two days before CIDR withdrawal. On the day of CIDR withdrawal (day 0), ewes received a 5mL intramuscular injection of PG-600 (400 IU equine chorionic gonadotropin and 200 IU human chorionic gonadotropin) or saline. Blood samples were collected before treatment and on the day of tissue collection to determine serum estradiol-17β and progesterone concentrations using radioimmunoassay. On days 4 and 7, six ewes from each treatment group were anaesthetised and a laparotomy was performed to obtain intercaruncular endometrial samples. Endometrial samples were collected ipsilateral to the ovary bearing the highest number of corpora lutea. An estradiol exchange assay was used to determine ESR1 concentration. Estradiol concentrations did not differ by time or treatment, but progesterone concentrations were significantly higher in the PG-600-treated group on day 7 despite a similar number of corpora lutea. Endometrial ESR1 concentration was significantly reduced on day 7 in the PG-600 group. In conclusion, although progesterone was higher than controls on day 7, ESR1 did not differ from controls suggesting that treatment with PG600 is unlikely to impair reproductive potential.

Effect of a progesterone-releasing intravaginal device (PRID) for 8 days during a modified Ovsynch protocol on pregnancy outcomes in lactating Holstein cows

Our objective was to evaluate the effect of a progesterone-releasing intravaginal device (PRID) in a 7-d Ovsynch protocol on pregnancy per artificial insemination (P/AI) and pregnancy loss, compared with a standard 7-d Ovsynch protocol without progesterone supplementation. We hypothesized that progesterone supplementation during an Ovsynch protocol would increase P/AI and decrease pregnancy loss. Data were collected on lactating Holstein cows (n = 716) that either received a 7-d Ovsynch protocol (control: d 0, 100 µg of GnRH; d 7, 500 µg of cloprostenol; d 9, µg of GnRH; n = 360) or a modified Ovsynch protocol with addition of a PRID (PRIDsynch; d 0, 100 µg of GnRH + PRID; d 7, 25 mg of dinoprost; d 8, PRID removal; d 9, 100 µg of GnRH; n = 356). All cows received timed artificial insemination (TAI) approximately 16 h after the second GnRH treatment. Pregnancy diagnosis was performed via ultrasonography on d 38 ± 3 after TAI and rechecked on d 80 ± 7 after TAI. Reproductive performance differed between treatments, with PRIDsynch cows having greater (38.9%) P/AI compared with control cows (31.7%) at d 38 ± 3 and also at d 80 ± 7 (34.6% vs. 28.9%, for PRIDsynch and control cows, respectively). Pregnancy loss did not differ among treatments.

Comparison of the 7-day CO-Synch and 8-day estradiol-based protocols for estrus synchronization and timed artificial insemination in suckled Bos taurus cows

Two experiments were conducted to compare, follicle diameter (FD) on Day −1, corpus luteum (CL) area on Day 7, progesterone (P4) concentration on Day 7 and 18, pregnancy per timed artificial insemination (TAI) on Day 30, and pregnancy loss (PL) between Days 30 and 60 after TAI (TAI, Day 0) using two different synchronization protocols. In Experiment 1, Angus cows (n = 1148) were randomly assigned to either 7-d progesterone CO-Synch (7-d CO-Synch) or 8-d progesterone + estradiol (8-d P + ES) synchronization protocols for TAI. On Day −10, cows in the 7-d CO-Synch treatment group (n = 574) received a progesterone-releasing intravaginal device (PIVD; 0.5 g P4) and GnRH (0.105 mg), on Day −3 the PIVD was removed and cows received cloprostenol (0.150 mg), then, on Day 0 (64 h after PIVD removal), cows received GnRH (0.105 mg) and were TAI. On Day −10, cows in the 8-d P + ES treatment group (n = 574) received a PIVD (0.5 g P4) and estradiol benzoate (2.0 mg), on Day −2 the PIVD was removed, and cows received cloprostenol (0.150 mg) and estradiol cypionate (0.5 mg), then, on Day 0 (48 h after PIVD removal), cows were TAI. Pregnancy per TAI was determined on Days 30 and 60. In a subset of cows (7-d CO-Synch, n = 41; 8-d P + ES, n = 40), serum P4 concentration was evaluated on Day 18. In Experiment 2, anestrus (n = 34) and cyclic (n = 34) suckled beef cows were selected and submitted at random on Day −10, to either 7-d CO-Synch or 8-d P + ES treatment groups. Follicle diameter on Day −1, CL area, and serum P4 concentration on Day 7 were determined. In Experiment 1, pregnancy per TAI on Day 30 did not differ (7-d CO-Synch = 48.9%; 8-d P + ES = 45.6%) between treatments but it was greater for cows with BCS ≥5 (P < 0.01). Pregnancy loss between Days 30 and 60 did not differ between treatment groups but tended to be greater in cows with BCS <5.0 (P < 0.1). In a subset of cows, serum P4 concentration on Day 18 did not differ between treatment groups but tended to be lower (P < 0.1) in cows that had PL between Days 30 and 60 compared to cows that had no PL. In Experiment 2, FD tended to be greater (P < 0.1) and CL area was greater (P = 0.05) in anestrus cows from 7-d CO-Synch treatment. In cyclic cows, the treatment did not affect the FD or CL area. In conclusion, there was no difference in pregnancy per TAI on Day 30 and PL between Days 30 and 60 between cows using 7-d CO-Synch + PIVD or 8-d estradiol-based + PIVD protocols for estrus synchronization and TAI.

126 Effect of oestrus synchronisation protocol with a high dose of oestradiol and an intravaginal progesterone-releasing device on the endometrial epidermal growth factor concentrations and fertility in recipient cows

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

PSV-B-15 Comparison of the 7-day Cosynch and 8-day Estradiol-Based Protocols for Synchronization of Ovulation and Timed Artificial Insemination in Suckled Bos Taurus Cows

Abstract The objective was to compare pregnancy rates (PR) on day 30 and pregnancy loss between days 30 and 60 (PL) after timed artificial insemination (TAI; day 0) using 2 synchronization protocols, and plasma progesterone on day 18 in suckled beef cows. Angus cows were randomly assigned to either 7-d progesterone CoSynch (7-d P + GnRH) or 8-d progesterone + estradiol (8-d P + ES) synchronization protocols for TAI. On day -10, cows in the 7-d P + GnRH treatment group (n=574) received a progesterone-releasing intravaginal device (PIVD) and GnRH, on day -3 the PIVD was removed and cows received cloprostenol. On day 0 (64 h after PIVD removal), cows received GnRH and were subjected to TAI. On Day -10, cows in the 8-d P + ES treatment group (n=574) received a PIVD and estradiol benzoate, on Day -2 the PIVD was removed, and cows received cloprostenol and estradiol cypionate. On day 0 (48 h after PIVD removal), cows were subjected to TAI. The TAI pregnancy rates for each treatment were determined on day 30 and day 60. In a subset of cows [n=81, (7-d P + GnRH, n=41; 8-d P + ES, n=40)], serum progesterone concentration was evaluated on day 18. The PR (7-d P + GnRH=48.9%; 8-d P + ES=45.6%) and PL (7-d P + GnRH=3.6%; 8-d P + ES=4.6%) did not differ (P≥0.28) between treatment groups. In a subset of cows, serum progesterone concentration on day 18 did not differ (P=0.67) between treatment groups but tended to be less (P=0.07) in cows that had PL compared with cows that had no PL. In conclusion, there was no difference in PR and PL between suckled beef cows using 7-d CoSynch or 8-d estradiol-based protocols for synchronization of ovulation and TAI.

171 Sex-Sorted Sperm Used for Fixed-Time Artificial Insemination and in Vitro Fertilization in Wood Bison

Abstract Wood bison (Bison bison athabascae) are a threatened species native to Canada. In a conservation setting such as zoos, females are preferred over males and sex-sorted sperm may help skew the proportion of females born. The objective of this study was to determine the efficacy of sex-sorted sperm in fixed-time artificial insemination (FTAI, Experiment 1) and in vitro fertilization (IVF, Experiment 2). In Experiment 1, females in bison herds at the University of Saskatchewan and Toronto Zoo were synchronized with 3.3 mg estradiol-17ß + 50 mg progesterone and given a progesterone-releasing intravaginal device for 8 days. At device removal, bison were given 500 µg of cloprostenol, and 72 h later were given 2500 IU human chorionic gonadotropin (hCG) and inseminated with either sex-sorted or conventional sperm. A second insemination occurred 24 h after hCG. Pregnancy diagnosis and fetal sexing was done by transrectal ultrasonography at 30- and 70-days post-AI, respectively. In Experiment 2, cumulus oocyte complexes collected transvaginally from live bison were fertilized in vitro with either sex-sorted or conventional sperm. Cleavage and embryo rates were determined at 3- and 7-days post fertilization, respectively. Proportional data were compared by chi-square. In Experiment 1, there was no effect of herd (P=0.17) or sperm (P=0.68) on pregnancy per AI (P/AI) at 30 days (total=25/51 [49%]). The proportion of female calves was greater in the sex-sorted than conventional group (9/10 [90%] vs 3/11 [27%]; P&amp;lt; 0.01). In Experiment 2, the cleavage (122/222 [55%] vs 62/94 [66%];P=0.07) and embryo rate (18/222 [8%] vs 14/94 [15%]; P=0.07) tended to be lower in the sex-sorted than the conventional group. In summary, sex-sorted sperm increased the proportion of female bison calves born following FTAI. Future efforts will include improving embryo production rates following IVF with sex-sorted sperm. Funding provided by Toronto Zoo, Mitacs, and STGenetics.

Comparison of the initial ovarian response, the synchrony of oestrus and ovulation and chronic stress response after administration of 100 or 250 μg of GnRH to randomly cycling Bos indicus cattle.

This study investigated the effects of administering saline, 100 or 250 μg of gonadotrophin releasing hormone (GnRH) on ovarian response, synchrony of oestrus and ovulation and chronic stress response in Bos indicus cattle. Randomised control. Animals were either left untreated (n=20) or on day 0 treated with an intravaginal progesterone releasing device and either saline (n=24), 100 μg (n=35), or 250 (n=35) μg of GnRH, intramuscular (IM). Blood was sampled 1.4 h after administration of treatment to monitor concentrations of luteinising hormone (LH) and P4 in serum and again 5 days later. On day 5 intravaginal P4 releasing device were removed, cloprostenol was administered IM and again 8 h later. Oestrus and ovulation were then monitored with ultrasonography for 6.5 days. Hair was clipped on day 55 for analysis of hair cortisol concentrations (HCC). No significant differences were found between Saline and GnRH treatments in the odds of inducing a new corpus luteum (CL) and the synchrony of oestrus or ovulation. HCC did not differ significantly between treatments. Mean concentrations of LH in serum on day 0 were less in the Saline compared to 100 and 250 μg GnRH treatments but did not differ between different doses of GnRH. Mean concentrations of LH and the odds of inducing a new CL were not increased after administering 250 μg compared to 100 μg of GnRH. Animal handling events in the study did not influence HCC. Further research is needed to better optimise responses to GnRH in B. indicus cattle.

Relationship between the concentration of anti-Müllerian hormone and antral follicle count in pasture-managed Holstein cows in the high tropics.

We evaluated the relationship between plasma levels of anti-Müllerian hormone (AMH) and the number of antral follicles at the restart of the follicular wave in crossbred Holstein cows reared under extensive grazing systems over 2500 m above sea level. The study included 140 cows from 15 farms that were in average at the 75.3 ± 2.10 d post partum. Animals were synchronized according to the following regime: day 0 = intravaginal progesterone releasing device (IPD) + estradiol benzoate (EB); day 7: withdrawal of IPD + prostaglandin; and day 8: EB, for restart of the follicular wave on day 11. On this day 11, antral follicle counts (AFCs) were made by transrectal ultrasound, and a plasma sample was taken for the determination of AMH. The mean AMH plasma level was 0.06 ± 0.03 ng/ml and the mean AFC was 17.26 ± 0.38 follicles. A strong positive linear correlation was found between these two variables (r = 0.783, r = 0.613, P < 0.0001). Cows were categorized according to AMH concentration as high (>0.09 ng/ml), intermediate (0.09-0.05 ng/ml) or low (<0.05 ng/ml). Cows with high AMH presented a higher AFC (25.0 ± 2.21 follicles) than those with low AMH (14.08 ± 2.68 follicles; P < 0.001. Our results suggest that the cut-off value of AMH = 0.09 ng/ml may be useful for selecting donors in multiple ovulation embryo transfer programs involving cows with these characteristics. Our data further suggest that AMH plasma concentration correlates with AFC and can be used as an endocrine biomarker of the number of antral follicles present at a given moment of the estrous cycle in crossbred Holstein cows raised at altitudes above 2500 m.

The relationship between circulating anti-mullarian hormone (AMH) and superovulatory response in buffaloes

The current study aimed to investigate the effect of using the superovulatory drug ‘follitropin’ on the follicular population and number of harvested buffalo embryos. At first, all animals were exposed to AMH analysis, Estrus was synchronized either by progesterone releasing intravaginal device (PRID) for 7 days or by double doses of cloprostenol sodium 11 days interval. A total dose of 40 IU of Follitropin was injected intramuscular for each animal as superovulatory drug (5 IU am and 5 IU pm for 4 consecutive days). AI was done using double straw/time for 3 times with 12 hours interval. Seven days later (day 0=ovulation day), embryos were recovered non-surgically. The results showed that total number of follicles/ animal was significantly increased in the group synchronized by PRID (7.4 ± 0.53) than the group synchronized by cloprostenol (4.8 ± 0.18), Number of the recovered embryos was significantly higher in the group synchronized by PRID (3.2 ± 0.71) than the group synchronized by cloprostenol (2.2 ± 0.37). In conclusion, AMH analysis is a crucial issue in expecting the superovulatory response in buffaloes, injection of follitropin as a superovulatory drug during diestrus phase in buffaloes that previously synchronized by PRID leads to a higher yielded embryo

Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus, pregnancy outcomes, and cost per pregnancy

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF2α - PRID; d 6, PGF2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d -6, GnRH + controlled internal drug release (CIDR); d -1, PGF2α - CIDR; d 0, PGF2α; d 2, GnRH + TAI]; (2) CIDR6 (d -6, GnRH + CIDR; d -1, PGF2α; d 0, PGF2α - CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.

Economic comparison of an ear tag automated activity monitor for estrus detection with timed-AI in Holstein heifers

The objective of this study was to compare the economic performance of an ear tag automated activity monitor system (AAM) versus a timed-AI (TAI) protocol in Holstein heifers. In total, 340 heifers were enrolled onto the study at 13.5 mo of age and randomly assigned to receive either an AAM (n = 170) or TAI (n = 170) protocol before breeding eligibility (D 0). Heifers in the AAM group were fitted with an ear tag AAM and bred based on high activity alert from the system. Heifers in the TAI group received a progesterone releasing intravaginal device on D −8, followed by device removal and prostaglandin on D −3 and gonadotropin-releasing hormone with TAI on D 0. In both treatments, the majority of heifers received sex-sorted semen for the first AI and conventional semen for subsequent AIs, with three opportunities to become pregnant. All heifers were diagnosed for pregnancy approximately 25 d post AI using transrectal ultrasonography, with confirmation at 30 and 45 d. Non-pregnant heifers in the TAI group, were resynchronized using the same TAI protocol. A partial budget was used to compare the costs and benefits of switching from a TAI to an AAM protocol in heifers, including protocol, labour, and rearing costs for each treatment, as well as estimated calf and milk value. Sensitivity analyses were also conducted to determine the effect of pregnancy per AI (P/AI), outsourcing AI, AAM tag cost and herd size on the net outcome. There was no difference in overall P/AI or days to pregnancy between treatments. However, number of AI was greater in the TAI than the AAM group. For the first AI, the P/AI was less in the TAI compared to the AAM group; however, the interval to first AI was less in TAI. There was minimal difference in performance for the second and third AI. There was a net gain of $11.97 per heifer when switching from a TAI to AAM protocol, due to the increased P/AI to the first AI and reduced cost of hormones. Several variables in the sensitivity analyses affected the net outcome. Considering only the first AI, switching to an AAM collar and a larger herd size all increased the net gain. Considering a greater P/AI to the first AI in the TAI group, outsourcing AI, using more AAM ear tags, and smaller herd sizes resulted in a net loss when switching from TAI to AAM. The AAM system resulted in exceptional P/AI and may be an economically viable alternative to improve heifer reproductive efficiency in herds with suboptimal P/AI to TAI.

Use of Intravaginal Progesterone-Releasing Device Results in Similar Pregnancy Rates and Losses to Long-Acting Progesterone to Synchronize Acyclic Embryo Recipient Mares.

The objectives of this study were: (1) to assess uterine features and serum progesterone concentrations of acyclic mares synchronized and resynchronized with intravaginal progesterone release device (IPRD), and (2) to compare pregnancy rates and losses of cyclic and acyclic embryo recipient mares treated with different synchronization protocols. In Experiment 1, mares (n = 12) received estradiol for 3 days (E2-3d), and then 24 h after the last injection, an IPRD was inserted and kept in place for 9 days. Three days after IPRD removal, mares were treated with E2-3d, and then a new IPRD was inserted and maintained for three days. Serum progesterone concentrations were assessed 2, 6, and 12 h after insertion and removal of IPRD, and then daily from the insertion of the first IPRD to one day after removal of the second IPRD. Experiment 2 was conducted with embryo recipient mares randomly assigned to four groups: (1) Cyclic: mares (n = 75) had ovulation confirmed after receiving a single dose of histrelin when a periovulatory follicle was first detected, (2) LAP4: acyclic mares (n = 92) were treated with E2-3d and then administered a single dose of LAP4 24 h after the last estradiol injection, (3) IPRD: acyclic mares (n = 130) were treated with E2-3d and an IPRD for 4–8 days, and (4) RE-IPRD: acyclic mares (n = 32) were synchronized as in the IPRD group but not used for embryo transfer (ET), then 8 to 15 days later, the mares were resynchronized with E2-3d and an IPRD for 4–8 days. In vivo-produced Day-8 embryos were collected and transferred 4–8 days after ovulation or progesterone treatments. Mares in IPRD and RE-IPRD groups had the intravaginal device removed immediately before ET, and then a new IPRD was inserted right after ET. Pregnancy diagnosis was performed at 5, 30, and 60 days after ET. Once pregnancy was confirmed, mares in the three acyclic groups received weekly injections of LAP4 (1.5 g) until 120 days of pregnancy. Mares in IPRD and RE-IPRD groups had the device removed three days after the first pregnancy diagnosis. In Experiment 1, progesterone concentrations increased rapidly starting 2 h after insertion of IPRD (p < 0.05); then, concentrations plateaued well above pregnancy maintenance until removal on days 9 and 3, respectively. Progesterone concentrations were reduced to baseline 24 h after IPRD removal (p < 0.05). For experiment 2, there was no difference in pregnancy rates across groups (65–74%) or pregnancy losses by 60 days of gestation (7–12%) (p > 0.05). In conclusion, the IPRD used herein resulted in a rapid increase and a sharp decline in progesterone concentrations upon its insertion and removal, respectively. Finally, our results demonstrated that IPRD could be a compatible alternative to LAP4 to synchronize and resynchronize acyclic embryo recipient mares.

Effect of ovulation synchronization program and season on pregnancy to timed artificial insemination in suckled beef cows

This study was conducted to (i) evaluate the requirement for the administration of GnRH coincident with insertion of a progesterone-releasing intravaginal device (PRID) and (ii) the effect of supplementing with 400 IU eCG at PRID removal on pregnancy per AI (P/AI) in spring and autumn calving suckled beef cows, subjected to a 7-d CO-Synch + PRID timed artificial insemination (TAI) program. Suckled beef cows (n = 1408) on 62 commercial farms were enrolled and randomly assigned to either of three treatments: 1) cows received a PRID and 100 μg GnRH on Day −10, followed by 25 mg PGF2α at PRID removal (Day −3) and 100 μg GnRH 72 h later (Day 0) at TAI (Treatment 1; n: spring = 236, autumn = 248); 2) as Treatment 1, but without GnRH at PRID insertion on Day −10 (Treatment 2; n: spring = 232, autumn = 227); 3) as Treatment 1, but cows also received 400 IU eCG at PRID removal on Day −3 (Treatment 3; n: spring = 233, autumn = 232). At Day −10, ovaries were examined by ultrasonography to evaluate the presence or absence of a corpus luteum (CL) and follicle(s) ≥ 10 mm in diameter. Body condition score (BCS) was assessed on a scale of 1–5. Pregnancy diagnosis was carried out 30–35 d after TAI by transrectal ultrasonography. Data were analyzed using the GENMOD and LOGISTIC procedures of SAS. There was a treatment by season interaction for P/AI (P < 0.001). In spring, overall P/AI was 59.1% (414/701) and was affected by treatment (59.3 v 49.6 v 68.2%, for Treatments 1, 2 and 3, respectively P < 0.05). In contrast, in autumn, overall P/AI (51.5%, 364/707) was unaffected (P > 0.05) by treatment (50.1 v 53.7 v 48.7% for Treatments 1, 2 and 3, respectively). Overall, eCG had a positive effect on P/AI for cows lacking a CL at treatment initiation (P < 0.05). In addition, in cows with low BCS (≤2.25), eCG supplementation tended (P = 0.09) to improve P/AI. Seasonal differences in response to synchronization treatment may be reflective of different management regimens (grazing v confinement) and breed type and remain to be elucidated.

Increasing conception rate by addition of a second prostaglandin injection in an Ovsynch and progesterone treatment programme for dairy cows not detected in oestrus before the start of the seasonal mating period

ABSTRACT Aims To assess the effect on conception rate to first service, 3- and 6-week in-calf rates, final pregnancy rate and the hazard of pregnancy, of addition of a second prostaglandin (PG) injection 24 hours after the first, in a synchrony programme combining Ovsynch and progesterone in pasture-based dairy cows that had not been detected in oestrus before the start of the seasonal mating period. Methods This prospective, negative-controlled study was conducted using anoestrous cows (n = 1,411) from nine spring-calving dairy herds in the Waikato region of New Zealand. Ovaries of cows not detected in oestrus were examined by transrectal ultrasonography and the presence of a corpus luteum determined. All cows were then treated with an intravaginal progesterone-releasing device and injected I/M with gonadorelin (Day −9). Cows were injected I/M with cloprostenol at device removal (Day −2) and on Day −1 were injected with either water (single PG; n = 720), or cloprostenol (double PG; n = 692). On Day 0 cows were injected again with gonadorelin, unless previously detected in oestrus, and were inseminated on Day 1. Based on dated pregnancy diagnosis by transrectal ultrasonography, cows were categorised as conceiving to first insemination, becoming pregnant in the first 3 or 6 weeks, or becoming pregnant by the end of the mating period. Results Cows in the double PG group had increased conception rate to first service (45.3 (95% CI = 45.1–45.4)% vs. 41.1 (95% CI = 41.0–41.3)%; p < 0.001), 3-week in-calf rate (53.4 (95% CI = 53.6–54.0)% vs. 49.2 (95% CI = 49.0–49.4)%; p < 0.001), and 6-week in-calf rate (67.2 (95% CI = 65.7-68.8)% vs. 63.5 (95% CI = 62.0-65.0)%; p = 0.014) compared to cows in the single PG group. Final pregnancy rate and hazard of pregnancy did not differ between treatment groups (p >0.2). Conclusions Addition of a second PG injection 24 hours after the first, increased conception rate to first service and the proportion of cows pregnant by 3 and 6 weeks after commencement of the mating period. Clinical relevance Improved outcomes can be achieved when treating those cows not detected in oestrus by the start of the seasonal mating period by addition of a second PG injection to the currently recommended treatment protocol for these animals.