Intravaginal Insert Research Articles

Embryo production by Holstein heifers superovulated with a recombinant long-acting follicle stimulating hormone analog.

This study aimed to evaluate embryo yield in Holstein heifers superovulated with a single injection of recombinant, long-acting human follicle-stimulating hormone (rFSH) versus multiple injections of pituitary-derived follicle-stimulating hormone (FSH). In Experiment 1, heifers were assigned randomly to one of 4 experimental groups: Control (280mg of pituitary derived FSH; six injections of 40mg and two injections of 20mg, each ~12h apart, n = 16); rFSH1 (50 μg of FSH analog protein, n = 16); rFSH2 (75 μg of FSH analog protein, n = 16) or; rFSH3 (100 μg of FSH analog protein, n = 16). The rFSH was administered as a single injection. Estrous cycles were presynchronized with gonadorelin acetate (GnRH) and an intra-vaginal progesterone insert (CIDR) on d -8, followed by cloprostenol sodium (PGF2α) on d -3 and d -2 with removal of the CIDR, and GnRH on d 0. On d 5, ovarian follicles ≥ 5mm in diameter were ablated by transvaginal ultrasound-guided aspiration. On d 6.5, heifers received a CIDR and the rFSH or the first injection of pituitary-derived FSH. On d 9, heifers received two injections of PGF2α, 12h apart. On d 10.5, the CIDR was removed, and on d 11, heifers received a GnRH injection. Heifers were AI-inseminated 12- and 24-hours post-GnRH injection, and uterine contents were flushed trans-cervically on day 18 (7 d after the GnRH injection). Ovarian follicles ≥ 5mm and corpora lutea were counted via ultrasound on days 5, 9, and 18. In Experiment 1, group did not affect (P = 0.52) the number of follicles ≥ 5mm (Control = 15.9 ± 1.2; rFSH1 = 17.5 ± 1.3; rFSH2 = 17.1 ± 1.3; rFHS 3 = 18.6 ± 1.4 follicles) or the number of corpora lutea (P = 0.96) on d 9 (Control = 1.1 ± 0.3; rFSH1 = 1.1 ± 0.3; rFSH2 = 1.1 ± 0.3; rFSH3 = 0.9 ± 0.2). Furthermore, there was no effect (P = 0.28) of rFSH dose on freezable embryos (grade 1 and 2 embryos) collected on d 18 (Control = 4.7 ± 1.1; rFSH1 = 4.7 ± 1.2; rFSH2 = 4.4 ± 1.1; rFSH3 = 2.6 ± 0.7 embryos). In Experiment 2, Control (n=8) and rFSH1 (n=16) groups were repeated in 3 replicates using the same protocols as Experiment 1. Consequently, Results showed that rFSH produced fewer total number of ova/embryos (Control = 9.9 ± 1.5 vs. rFHS1 = 5.9 ± 0.9, P = 0.04) and fewer freezable embryos (Control = 5.3 ± 1.0 vs. rFSH1 = 1.4 ± 0.3, P < 0.01). In conclusion, the single rFSH injection effectively induced superovulation; however, its repeated use reduced embryo production.

Effect of the Application of PGF2α Associated With Ovulation Induction in a Fixed-Time Superovulation Programme for Precocious Nellore Heifers.

This study evaluated the effect of prostaglandin F2α (PGF2α) associated with gonadotropin-releasing hormone (GnRH) for ovulation induction in precocious indicus heifers submitted to a fixed-time superovulation (SOV) programme. Precocious Nellore heifers (n = 35), aged 13 months, were subjected to the SOV protocol. On day 0 (D0), all animals received intravaginal insertion of a progesterone (P4) device along with intramuscular administration of 2 mg of oestradiol benzoate, plus 200 IU of follicle-stimulating hormone in decreasing doses, with 12-h intervals between D4 and D7, in addition to 150 μg of D-cloprostenol on D6 and device removal on D7. On D8, the donors received 10.5 μg of buserelin acetate and the treatment group received 300 μg of D-cloprostenol/PGF2α. Artificial insemination was performed 12 h and 24 h after GnRH administration using frozen semen. On D15 of the protocol (i.e., D7 after insemination), the embryos were collected and evaluated. All animals passed through the control and treatment groups. Results were evaluated by analysis of variance using an adjusted mixed-effects model (p < 0.05). There was no difference in the total number of embryos between the control and treatment groups (10.40 ± 1.52 vs. 9.60 ± 1.36; p = 0.63) or viable embryos (6.30 ± 1.22 vs. 4.30 ± 0.71). For precocious indicus heifers, treatment with PGF2α in association with GnRH did not affect embryo production in the fixed-time SOV protocol.

Optimizing ReBreed21 II: Fertility and reproductive efficiency in different parities during a shortened breeding season in beef cattle

This study evaluated the efficiency of a rapid reinsemination program allowing timed AI (TAI) every 21d (ReBreed21) in a commercial beef cow-calf operation. Nelore females from different parities (n = 2085) were synchronized for first TAI (D0 = TAI) using an estradiol/progesterone (E2/P4) protocol and assigned to one of three reinsemination programs: Resynch33 (n = 753), traditional resynch program with second TAI at D42 after first TAI; ReBreed21 (n = 687); or ReBreed21+EC (n = 670). The ReBreed females (n = 1357) received intravaginal P4 insert on D12, on D19 P4 was removed, and a dose of equine chorionic gonadotropin (eCG) was administered, then, ReBreed21 females received 0.6 mg of EC (ReBreed21+EC) or nothing (ReBreed21) and on D21, nonpregnancy (NP) was determined using Doppler ultrasound to detect corpus luteum (CL) blood flow (BF) (NP: <25 % BF pixels of total CL area) and NP cows received immediate TAI and GnRH to induce ovulation. Pregnancy diagnosis was performed at D33 after TAI following all TAIs. Cows considered pregnant at D21, based on CL BF, but NP on D33 were designated False-Positives (FP) and false negatives (FN) were number of nonpregnant cows/heifers on d21 based on the CL BF found to subsequently be pregnant on D33 divided by the total number pregnant. Pregnancy/AI (P/AI) did not differ for the first TAI (55.1 %) among the treatments. Heifers had similar P/AI at the second AI in all groups and similar to the first AI. Primiparous had greater P/AI in ReBreed21+EC and Resynch33 at s TAI compared to ReBreed21, 51.7 %, 55.8 %, 34.2 %, respectively. Multiparous had greater P/AI at second TAI in Resynch33 (60.9 %) than ReBreed21 programs (34.7 %). The percentage FP and FN among ReBreed21 programs did not differ, 13.8 and 0.2 %, respectively. Overall accumulative pregnancies on D21 of the breeding season were greater for ReBreed21 and ReBreed21+EC than Resynch33 (69.7 %, 71.6 %, and 55.5 %, respectively). However, on D42 of the breeding season, only heifers had greater pregnancies in ReBreed21 programs than Resynch33 (73.3 %, 74.3.6 %, and 63.2 %, respectively). Average days to pregnancy were less (P = 0.01) for ReBreed21 and ReBreed21+EC than Resynch33. Thus, the ReBreed21 strategy can improve the efficiency of TAI programs in beef cattle. Of interest, ReBreed21 was particularly effective in nulliparous, somewhat effective in primiparous when EC was added to the program, but relatively ineffective in multiparous beef cattle.

Biochemical profile and oocyte quality of primiparous Bos indicus cows submitted to a timed artificial insemination protocol

The objective of this study was to verify the causes of the lower response of primiparous Bos indicus cows to the ovulation synchronization protocol. Two experiments were performed to evaluate the biochemical profile, oocyte and follicular cell quality (Experiment 1) and pregnancy (Experiment 2). In Experiment 1, suckled primiparous (n = 24) and multiparous cows (n = 24) were submitted to ovum pick up (OPU). On Day 0 (D0), all cows received 2 mg of estradiol benzoate (EB) and an intravaginal progesterone insert (P4). Five days (D5) after the first hormonal administration (EB + P4), all follicles larger than 3 mm were counted on each ovary, and ovum pick-up (OPU) was performed. On day 12 (D12), the intravaginal progesterone insert was removed, and measurement and aspiration of the largest follicle were performed. Blood samples were collected on D5 and D12 to evaluate the concentrations of glucose, cholesterol, NEFA, IGF-1 and insulin. In Experiment 2, suckled primiparous (n = 50) and multiparous (n = 50) cows were subjected to an ovulation synchronization protocol based on E2/P4 (D0: 2 mg EB plus P4 intravaginal insert; D8: 500 μg of cloprostenol, 1 mg cypionate estradiol and 300UI of eCG; D10: artificial insemination). In addition, blood samples were collected on D10 for evaluation of the same hormones and metabolites described in Experiment 1. In all studies, calves remained with the cows during the experimental period. In experiment 1, the number of oocytes grade 1 (P = 0.83), grade 2 (P = 0.23) and grade 3 (P = 0.51), total number of retrieved oocytes (P = 0.14), oocyte quality index (P = 0.93) and total viable oocytes (P = 0.38) did not differ between primiparous and multiparous cows. The number of follicles at the time of follicular aspiration (20.7 ± 1.5 vs. 18.0 ± 1.9; P = 0.05) and the diameter of the largest follicle on D12 (13.5 ± 0.6 vs. 11.4 ± 0.6; P = 0.04) were greater in multiparous cows, and the number of degenerated oocytes was greater in primiparous cows (1.9 ± 0.7 vs. 1.2 ± 0.3; P = 0.05). On D5, the concentrations of IGF-1 (P = 0.47), insulin (P = 0.08), total cholesterol (P = 0.47), NEFA (P = 0.77) and glucose (P = 0.55) in the blood and IGF-1 (P = 0.97) and insulin (P = 0.11) in the follicular fluid did not differ between parity groups. On D12, there was no difference in the concentrations of IGF-1 (P = 0.26), total cholesterol (P = 0.32), NEFAs (P = 0.31) and glucose (P = 0.93) in the blood between primiparous and multiparous cows, however, the serum insulin concentration (P = 0.04) was higher in primiparous cows. There was no correlation between serum and follicular fluid insulin concentrations (r = 0.17; P = 0.31), however, there was a low correlation between serum and follicular fluid IGF-1 concentrations (r = 0.47; P = 0.002). Quantification of transcripts did not differ between parity groups. In experiment 2, concentrations of NEFA (P = 0.12) and insulin (P = 0.16) in the blood and P/AI (P = 0.93) did not differ between parity [60 % (30/50) primiparous vs. 60 % (30/50) multiparous]. In contrast, blood concentrations of IGF-1 (P = 0.0001), total cholesterol (P = 0.005) and glucose (P = 0.01) were greater in primiparous cows. It was concluded that the oocyte quality and expression of the genes evaluated in the granulosa cells were not different between primiparous and multiparous cows. Unexpectedly, the pregnancy rate did not differ between parity. Nevertheless, the blood concentrations of IGF-1, total cholesterol and glucose were greater in primiparous cows.

Effect of 200 μg of gonadorelin hydrochloride at the first GnRH of a CO-Synch program on ovulation rate and pregnancies per artificial insemination in Holstein heifers

The initial ovulatory response during synchronization programs is often low in dairy heifers, largely due to follicular dynamics and hormonal dynamics. Specifically, the progesterone (P4) concentration at the time of the first GnRH treatment in a breeding program can influence the LH response, often resulting in a suboptimal ovulatory response. The objective of this study was to determine the effect of the highest label dose 200 μg (100 μg vs. 200 μg) of GnRH (50 μg of gonadorelin hydrochloride per mL; Factrel, Zoetis Inc. Madison, NJ) at the first GnRH of a 6-d CO-Synch plus P4 device program on ovulatory response and pregnancy per AI (P/AI) in first service in Holstein heifers. A total of 1,308 Holstein heifers were randomly allocated at the beginning of a 6-d CO-Synch program at day 0 to receive either i.m. treatment of 100 μg (2CC, n = 655) or 200 μg (4CC, n = 653) of GnRH. Also, at d 0, heifers received an intravaginal insert with 1.38 g of P4 (Eazi-Breed CIDR Cattle Insert, Zoetis Inc.). On day 6, the insert was removed, and i.m. treatment of 25 mg of PGF2α (12.5 mg of dinoprost tromethamine/mL; Lutalyse HighCon Injection, Zoetis Inc.) was administered. On d 7, a second i.m. treatment of 25 mg of PGF2α was given, followed on d 9 by concurrent i.m. treatment of 100 μg of GnRH, and timed AI. A subset of 396 heifers had their ovaries scanned to evaluate ovulatory response, and blood samples were collected to measure the serum concentration of P4 at d 0 and d 6 of the study. The P4 concentrations at d 0 were categorized as low (≤3 ng/mL) or high (>3 ng/mL). The ovulatory response was greater for heifers receiving 4CC than 2CC at d 0 (54.7% vs. 42.8%). The ovulatory response was greater for low P4 than high P4 at d 0 (54.3% vs. 37.8%). However, we did not observe an interaction between treatment and P4 concentrations (low P4 2CC = 48.6% vs. high P4 2CC = 30.0%; low P4 4CC = 60.0% vs. high P4 4CC = 45.5%). The receiver operating characteristic curve analysis indicated that P4 concentrations at d 0 treatment could predict the ovulatory response, although the area under the curve was only 0.6. As expected, heifers that ovulated had increased P/AI (no = 55.6% vs. yes = 67.7%); however, we found no effect of treatment on P/AI (2CC = 63.3% vs. 4CC = 59.6%), and no interactions between treatment and ovulation and treatment and P4 (high vs. low) for pregnancy outcomes. In summary, P4 concentration and increasing the dose of GnRH at d 0 positively affected ovulatory response in Holstein heifers. However, there was no interaction between treatment and P4 on ovulation and no subsequent impact of GnRH dose on P/AI.

(124) An Open-Label, Vulvoscopic Pilot Study of Intravaginal Prasterone in Menopausal Women with Dyspareunia Examining Vestibular Tissue Changes

Abstract Introduction Prasterone (DHEA), a daily 6.5 mg intravaginal insert, is approved in the US and Canada for moderate to severe dyspareunia, a symptom of vulvovaginal atrophy from menopause. Prasterone converts intracellularly in vaginal epithelial cells into active androgens and estrogens without affecting systemic hormone blood levels. Although prasterone is a vaginal insert, the significant reduction of pain in the phase 3 registration trials suggests improvement of the vestibular endodermal genital tissue health associated with dyspareunia in androgen and estrogen deficient states like menopause. To the best of our knowledge, there have not been previous prospective studies documenting visible changes to the vestibule with daily administration of 6.5 mg prasterone in menopausal women with vulvovaginal atrophy and moderate to severe dyspareunia. Objective It is the aim of this study to investigate whether an intravaginal therapy, prasterone, could affect the vestibular tissue, assessing for changes in visual appearance, pain, and responses on sexual event diaries over the 20-week administration of intravaginal 6.5 mg prasterone. Methods This 20-week, open-label, prospective, pilot study included 11 menopausal women (median age = 56 y) treated daily with 6.5 mg prasterone intravaginal inserts and assessed every 4 weeks. During vulvoscopy, vestibular pain was assessed by cotton-tipped swab testing and vestibular and vaginal health were independently assessed using the Visual Scale (VS). In addition, vulvoscopic photographs were obtained and assessed using the Vulvoscopic Genital Tissue Appearance (VGTA) scale to evaluate overall genital tissue health. Mean changes from baseline for genital tissue health and pain assessments were analyzed by repeated measures one-way analysis of variance, followed by Dunnett’s post-hoc test. Sexual event diaries were completed, and adverse events recorded. Results Eleven participants completed the study, with a total of 1607 vulvoscopic photographs, showing reduction in vestibular erythema and pallor at end of study. Total VGTA scores at week 20 showed improvement from baseline (mean change = -7.9, p = 0.0016). Changes in VS scores were significantly reduced from baseline after active treatment by week 4 and maintained through week 20 for the vestibule (3.00, p = 0.004) and vagina (4.00, p = 0.002) respectively), suggesting improved vestibular and vaginal tissue health. Cotton-tipped swab test scores were significantly reduced from baseline after week 8 through week 20 (mean change = (-7.27, p = 0.019), suggesting decreased pain. Diary results showed decrease in pain from 81.3% during the first month to 8.3% during the last month of the study, with sexual activities discontinued due to discomfort reduced from 45.8% to 6.3% respectively. No prasterone related adverse events were reported. Conclusions This is the first prospective vulvoscopic study showing that intravaginal hormone therapy significantly improves total VGTA, VS and cotton-tipped swab test scores. This observation may explain the robust effect of prasterone in lowering dyspareunia in menopausal women with moderate to severe dyspareunia. Our findings suggest that intravaginal prasterone exerts biologic activity on the androgenic endodermal vestibule, as the medication passes from vagina to vestibule, resulting in amelioration of pain associated with sexual activity. Disclosure Yes, this is sponsored by industry/sponsor: AMAG Pharmaceuticals, Inc. Clarification: Industry funding only - investigator initiated and executed study.

Influence of repeated estrus synchronization treatment on hormone secretion, growth, and development of dairy goats.

In large-scale intensive farms, dairy goats often undergo frequent estrus synchronization (ES) treatment, which may result in a decline in reproductive performance; however, the underlying mechanism remains unclear. The present study aimed to investigate the effect of pregnant mare serum gonadotropin (PMSG) and progesterone (P4)-mediated ES treatment on fertility in dairy goats, while also identifying key metabolic and endocrine mechanisms that influence reproductive performance in does subjected to repeated ES treatment. Forty-eight Saanen does were randomly assigned to two groups (24 goats each) that received ES treatments either thrice fortnightly (3-PMSG) or once (1-PMSG) simultaneously with the third ES treatment of the 3-PMSG group during the breeding season. ES treatment was performed via the intravaginal insertion of a controlled internal drug release (CIDR) device impregnated with 300 mg P4, followed by 300 IU PMSG injections 48 h before CIDR withdrawal. Blood was collected to detect the level of hormones and blood biochemical indices. Additionally, estrus rate, fecundity rate, body weight, size, and lactation performance were measured. The results showed that repeated ES treatment markedly decreased the estrus rate and fecundity rate of goats. Among the does in all groups, there was no substantial difference in follicle stimulating hormone, luteinising hormone, gonadotropin-releasing hormone, melatonin, growth hormone, PMSG, total cholesterol, total protein, and glucose levels, as well as the body weight, body size, and lactation performance. Repeated ES treatment elevated estrogen (E2) levels 36, 48, and 72 h post-CIDR removal; increased P4 upon CIDR insertion; and raised PMSG antibody levels 24, 48, and 72 h post-CIDR removal. The results suggest that elevated anti-PMSG levels are the primary reason for the decline in ES efficiency, and that high E2 and P4 levels at some time points also impair reproductive performance. These findings provide novel insights into the metabolic effects of repeated PMSG stimulation in goats, guiding future reproductive hormone use in breeding practices.

Inducing luteolysis prior to atresia of the first follicular wave by altering the interval to prostaglandin F2ɑ administration after long-term progestin presynchronization

A series of experiments was designed to evaluate treatment schedules for control of the estrous cycle in which luteolysis is induced prior to atresia of the first follicular wave following a long-term progestin presynchronization treatment in beef heifers. The overarching hypothesis was that the proportion of heifers undergoing luteolysis and expressing estrus following PG administration would be affected by the duration of treatment with an intravaginal progesterone-releasing insert (CIDR; 1.38 g progesterone) and/or by the interval from CIDR removal to prostaglandin F2ɑ (PG; 500 μg cloprostenol) administration. In each experiment, heifers were blocked by reproductive tract score (RTS) and body weight (BW) and were assigned randomly within block to treatment. In Experiment 1, heifers (n = 91) were assigned to one of four treatments in a 2 x 2 factorial design: presynchronization via CIDR treatment for either 14 d or 18 d with administration of PG either 8 d or 9 d following CIDR removal. In Experiment 2, heifers (n = 63) were assigned to receive administration of PG either 9 d or 10 d following a 14 d CIDR presynchronization. In Experiment 3, heifers (n = 83) were assigned to receive administration of PG either 9 d or 16 d following a 14 d CIDR presynchronization. In Experiment 1, there was a tendency for a greater proportion of heifers to express estrus by 96 h after PG administration when PG administration occurred 9 d versus 8 d after CIDR removal (P = 0.07). Additionally, serum estradiol (E2) concentrations were greater at the time of PG administration when PG was administered 9 d versus 8 d after CIDR removal following an 18 d CIDR treatment (P = 0.006). In Experiment 2, pregnancy rate to AI (P/AI) was greater among heifers receiving PG administration 9 d versus 10 d after CIDR removal (P = 0.03). There was a tendency for largest follicle diameter (LFD) to be greater among non-estrous heifers receiving PG administration 10 d versus 9 d after CIDR removal when measured at timed AI (P < 0.08). In Experiment 3, there was a tendency for serum progesterone (P4) concentrations at the time of PG administration to be greater among heifers receiving PG administration 16 d versus 9 d after CIDR removal (P = 0.07). These results provide a preliminary evaluation of the feasibility of inducing luteolysis prior to atresia of the first follicular wave following long-term progestin presynchronization.

Evaluation of later timepoints for fixed-time artificial insemination of beef heifers and cows when using sex-sorted semen

Two experiments were designed to evaluate later timepoints for fixed-time artificial insemination (FTAI) of beef heifers and cows, with the hypothesis that use of a later timepoint would allow a greater proportion of animals to express estrus prior to FTAI and result in greater conception rates among estrous females inseminated with sex-sorted semen. In Experiment 1, estrus was synchronized for 1640 heifers using the 14 d CIDR-PG protocol: insertion of an intravaginal progesterone-releasing insert (CIDR; 1.38 g progesterone) on Day −33 and removal on Day −19, and administration of prostaglandin F2α (PG; 500 μg cloprostenol sodium) on Day −3. Heifers were inseminated at one of three FTAI timepoints: 66 h, 70 h, or 74 h after PG administration. In Experiment 2, estrus was synchronized for 414 beef cows using the 7 & 7 Synch protocol: administration of PG coincident with CIDR insertion on Day −17, gonadotropin-releasing hormone (GnRH; 100 μg gonadorelin) on Day −10, and PG coincident with CIDR removal on Day −3. Cows were inseminated at one of two FTAI timepoints: 66 h or 72 h after PG administration. In both experiments, only animals that expressed estrus prior to FTAI were inseminated with sex-sorted semen. In Experiment 1, the proportion of heifers that expressed estrus prior to FTAI (66 h: 62 %; 70 h: 67 %; 74 h: 71 %) was greater when FTAI was performed at 74 h versus 66 h (P = 0.0097); however, conception rate of heifers that expressed estrus and were serviced with sex-sorted semen did not differ among treatments (P = 0.67; 66 h: 56 %; 70 h: 53 %; 74 h: 53 %). In Experiment 2, the proportion of cows expressing estrus prior to FTAI did not differ between treatments (P = 0.30; 66 h: 71 %; 72 h: 76 %). Additionally, conception rate of estrous cows inseminated with sex-sorted semen did not differ between treatments (P = 0.24; 66 h: 45 %; 72 h: 40 %). These results indicate that performing FTAI later following the 14 d CIDR-PG protocol increases the proportion of heifers that express estrus and are serviced with sex-sorted semen but does not improve conception rates. Later timing of FTAI following the 7 & 7 Synch protocol was not observed to increase the proportion of cows expressing estrus prior to FTAI or improve conception rates among estrous cows inseminated with sex-sorted semen. Together, these results provide further insight into optimal timing of FTAI when using sex-sorted semen.

PSXIV-24 Inducing Luteolysis Before Atresia of the First Follicular Wave By Altering the Interval to Prostaglandin F2ɑ Administration After Long-Term Progestin Presynchronization

Abstract A series of experiments was designed to evaluate treatment schedules for control of the estrous cycle in which luteolysis is induced before atresia of the first follicular wave following a long-term progestin presynchronization treatment in beef heifers. The central hypothesis was that the proportion of heifers undergoing luteolysis and expressing estrus following PG administration would be affected by the duration of treatment with an intravaginal progesterone-releasing insert (CIDR; 1.38 g progesterone) and/or by the interval from CIDR removal to prostaglandin F2ɑ (PG; 500 µg cloprostenol) administration. In each experiment, heifers were blocked by reproductive tract score and body weight and assigned randomly within block to treatment. In Experiment 1, heifers (n = 91) were assigned to one of four treatments in a 2 x 2 factorial design: presynchronization via CIDR treatment for either 14 d or 18 d with administration of PG either 8 d or 9 d following CIDR removal. In Experiment 2, heifers (n = 63) were assigned to receive administration of PG either 9 d or 10 d following a 14 d CIDR presynchronization. In Experiment 3, heifers (n = 83) were assigned to receive administration of PG either 9 d or 16 d following a 14 d CIDR presynchronization. In Experiments 1 and 2, heifers were artificially insemination (AI) 8 to 12 h after expressing estrus, with timed AI performed 96 h after PG administration for heifers failing to express estrus. In Experiment 3, heifers expressing estrus by 66 h after PG administration were inseminated at that time and all remaining heifers were inseminated at 90 h. In Experiment 1, there was a tendency for a greater proportion of heifers to express estrus by 96 h after PG administration when PG administration occurred 9 d versus 8 d after CIDR removal (P = 0.07). Additionally, serum estradiol concentrations were greater at the time of PG administration when PG was administered 9 d versus 8 d after CIDR removal following an 18 d CIDR treatment (P = 0.006). In Experiment 2, pregnancy rate to AI (P/AI) was greater among heifers receiving PG administration 9 d versus 10 d after CIDR removal (P = 0.03). There was a tendency for largest follicle diameter measured at timed AI to be greater among non-estrous heifers receiving PG administration 10 d versus 9 d after CIDR removal (P &amp;lt; 0.08). In Experiment 3, there was a tendency for serum progesterone concentrations at the time of PG administration to be greater among heifers receiving PG administration 16 d versus 9 d after CIDR removal (P = 0.07). These results provide a preliminary evaluation of the feasibility of inducing luteolysis before atresia of the first follicular wave following long-term progestin presynchronization.

292 Tandem Administration of Prostaglandin F2α and Gonadotropin-Releasing Hormone in Beef Heifers and Cows as a Convergent Presynchronization Method in the 7 &amp; 7 Synch Protocol

Abstract An experiment was designed to evaluate modifications to the 7 &amp; 7 synch protocol with the hypothesis that tandem administration of prostaglandin F2α (PG) and gonadotropin-releasing hormone (GnRH) during the presynchronization portion of the protocol would generate a convergent ovarian response among anestrous and estrous cycling females. Nulliparous (n = 153), primiparous (n = 92), and multiparous (n = 413) Angus-cross females across four locations were blocked based on parity and pre-treatment cyclicity status (presence or absence of a corpus luteum) as assessed via transrectal ovarian ultrasonography. Within block, females were randomly assigned to one of three treatments. Females assigned to the 7 &amp; 7 Synch treatment received an intravaginal progesterone-releasing insert (CIDR) and administration of PG on day 0, administration of GnRH on day 7, and administration of PG coincident with CIDR removal on day 14. Females assigned to 7 &amp; 7 Synch + G received the same treatment schedule with the modification that GnRH was administered in tandem with the PG on day 0. Females assigned to 7 &amp; 7 Synch + G + P received GnRH in tandem with PG on both day 0 and day 7 of the treatment schedule. Fixed-time artificial insemination (FTAI) was performed at 54 h after CIDR removal for nulliparous females and at 66 h for primiparous and multiparous females. Transrectal ovarian ultrasonography was performed on day 7 and day 14 to characterize the largest follicle diameter (LFD) as well as the presence/absence and number of corpora lutea (CL). Expression of estrus before FTAI was characterized in three locations using estrus detection patches (Estrotect. Approximately 75 to 90 days after FTAI, pregnancy status was determined via transrectal ultrasonography. Day 7 CL status was affected by treatment (P = 0.002), with a greater proportion of females presenting with CL on Day 7 when treated with the modified treatments (64%) as compared with the 7 &amp; 7 Synch treatment (31%). Day 14 LFD was affected by treatment (P = 0.002), with a larger average LFD among females treated with 7 &amp; 7 Synch + G + P (12.0 mm) as compared with 7 &amp; 7 Synch (11.4 mm) or 7 &amp; 7 + G (10.7 mm). Pregnancy rate to FTAI was affected by pre-treatment cyclicity status (P &amp;lt; 0.0001) and semen type (P = 0.0003) and tended to be affected by treatment (P = 0.09) with 7 &amp; 7 Synch + G + P tending to result in a greater proportion pregnant (58.86%) as compared with 7 &amp; 7 Synch (49.17%) and 7 &amp; 7 Synch + G (48.79%). Tandem administration of GnRH and PG on days 0 and 7 of the 7 &amp; 7 Synch protocol effectively generated a convergent ovarian response among anestrous and estrous cycling beef females and tended to result in increased pregnancy rates to FTAI.

In Vitro, In Compost, and In Vivo Assessment of Chitosan-Polyethylene Glycol as an Intravaginal Insert for Progesterone Delivery in Sheep

In Vitro, In Compost, and In Vivo Assessment of Chitosan-Polyethylene Glycol as an Intravaginal Insert for Progesterone Delivery in Sheep

Vestibular tissue changes following administration of intravaginal prasterone: a vulvoscopic open-label pilot study in menopausal women with dyspareunia.

Prasterone, an intravaginal dyspareunia treatment in menopausal women, improves vaginal health through intracellular conversion of dehydroepiandrosterone into androgens and estrogens. Phase 3 trials for prasterone showed significant improvement in vaginal tissue health and reduction of pain. To assess vestibular changes with daily use of intravaginal prasterone in menopausal women with moderate to severe dyspareunia. This open-label prospective pilot study was conducted over 20 weeks. It included 11 menopausal women (median age,56 years) who were treated daily with intravaginal inserts of 6.5-mg prasterone and assessed monthly. During vulvoscopy, vestibular pain was assessed by cotton-tipped swab testing, and vestibular and vaginal health was independently assessed with the Visual Scale (VS). In addition, vulvoscopic photographs were obtained and assessed via the Vulvoscopic Genital Tissue Appearance (VGTA) scale to evaluate overall genital tissue health. Mean changes from baseline for genital tissue health and pain assessments were analyzed by repeated measures 1-way analysis of variance, followed by a Dunnett post hoc test. Sexual event diaries were completed and adverse events recorded. Outcomes included indices of genital tissue health: pain assessment by cotton-tipped swab testing, VS of the vestibule and vagina, VGTA, and sexual event diary. Aggregate scores from the cotton-tipped swab test progressively improved, reaching statistical significance at week 16, which was maintained through week 20 (-7.27, P= .019). VS scores significantly improved from baseline by week 4 and were maintained through week 20 for the vestibule (-3.00, P= .004) and vagina (-4.00, P= .002). An overall 1607 vulvoscopic photographs were examined; all showed reduction in vestibular erythema and pallor at the end of the study. The mean change from baseline at week 20 for the VGTA score was -7.9 (P= .0016). Intercourse associated with pain was reduced from 81.3% of initiated events during the first month of the study to 8.3% during the last month. Sexual activities that were discontinued due to discomfort were reduced from 45.8% to 6.3%. No prasterone-related serious adverse events were reported. Prasterone, a safe and effective intravaginal hormone treatment, significantly improves vestibular health parameters. Strengths are the prospective study design and the use of multiple outcome measures to assess vestibular tissue health and pain associated with sexual activity. Limitations are the small study cohort and use of nonvalidated outcome measures. Our findings suggest that intravaginal prasterone exerts biologic activity on the androgenic endodermal vestibule, as the medication passes from vagina to vestibule, resulting in amelioration of pain associated with sexual activity.

Role of aquaporin 3 in reproductive performance of dairy goats after repeated estrus synchronization stimulation.

This study explored the specific molecular mechanisms through which repeated estrus synchronization (ES) treatments reduce the reproductive performance of dairy goats. Ninety-six goats (n=24/group) were randomly assigned to two groups receiving ES treatments thrice every fortnight (3- equine chorionic gonadotropin [eCG] and 3-follicle stimulating hormone [FSH] groups) and two groups receiving one ES treatment (1-eCG and 1-FSH groups). ES treatments of 1- and 3-eCG goats were performed via the intravaginal insertion of a controlled internal drug release (CIDR) device containing 300 mg progesterone (P4), followed by 300 IU eCG injections 48 h before CIDR withdrawal. The 1- and 3-FSH goats received CIDR for 10 d, followed by 50 IU FSH and 100 μg PGF2α within 12 h of CIDR withdrawal. Ovaries of three goats in estrus from both groups were harvested for analysis. Subsequently, all the goats in estrus were artificially inseminated twice. Consequently, 3-eCG and 3-FSH goats showed a considerably reduced estrus rate and litter size than 1-eCG and 1-FSH goats. AQP3 mRNA and protein expression were significantly higher in the 3-eCG and 3-FSH groups than in the 1-eCG and 1-FSH groups. AQP3 overexpression led to cell apoptosis and decreased steroid hormone secretion ability of ovarian granulosa cells. Moreover, it resulted in a decrease in maturation and cleavage rates after parthenogenetic activation and in vitro fertilization, respectively. AQP3 gene was involved in reducing the reproductive performance of repeated ES-treated dairy goats. These findings provide a theoretical foundation for the effective use of reproductive hormones in breeding techniques for livestock.

Effect of Vaginal Washing before Intravaginal Prostaglandin Insertion on Induction of Labor: A Systematic Review and Meta-analysis of Randomized Controlled Trials.

Saline vaginal douching prior to intravaginal prostaglandin application might increase the vaginal pH, leading to improvement of prostaglandin bioavailability, by which the outcomes of labor induction can be greatly improved. Thus, we aimed to evaluate the effect of vaginal washing with normal saline before insertion of vaginal prostaglandin for labor induction. A systematic search was done in PubMed, Cochrane Library, Scopus, and ISI Web of Science from inception to March 2022. We selected randomized controlled trials (RCTs) that compared vaginal washing using normal saline versus no vaginal washing in the control group before intravaginal prostaglandin insertion during labor induction. We used RevMan software for our meta-analysis. Our main outcomes were the duration of intravaginal prostaglandin application, duration from intravaginal prostaglandin insertion to active phase of labor, duration from intravaginal prostaglandin insertion till total cervical dilatation, labor induction failure rate, incidence of cesarean section (CS), and rates of neonatal intensive care unit (NICU) admission and fetal infection postdelivery. Five RCTs were retrieved with a total number of 842 patients. Duration of prostaglandin application, duration from prostaglandin insertion to active phase of labor, and time interval from prostaglandin insertion to total cervical dilatation were significantly shorter among vaginal washing group (p < 0.05). Vaginal douching prior to prostaglandin insertion significantly decreased the incidence of failed labor induction (p < 0.001). After the removal of reported heterogeneity, vaginal washing was linked to a significant decline in CS incidence (p = 0.04). In addition, the rates of NICU admission and fetal infection were significantly lower in the vaginal washing group (p < 0.001). Vaginal washing with normal saline before intravaginal prostaglandin insertion is a useful and easily applicable method for labor induction with good outcomes. · Induction of labor is frequently used in the obstetrics field.. · We assessed vaginal washing impact before prostaglandin insertion for labor induction.. · Vaginal washing is an easily applicable method for labor induction with good outcomes..

Modern integrated approach in the treatment of atrophic vaginitis in women of menopausal age

The objective: to evaluate the clinical effectiveness of treatment of atrophic vaginitis in menopausal women using combined therapy, which includes vaginal suppositories with hyaluronic acid. Materials and methods. A clinical and laboratory examination of 60 women 50-65 years old with atrophic vaginitis was carried out. The main group (30 patients) received combined therapy: a combination of local estrogen with intravaginal insert of a drug with hyaluronic acid. The patients of the control group (30 women) received only local therapy with estrogens.Results. Clinical and laboratory remission after the treatment was determined in 100 % of women in the main group and 63.3 % of patients in the control group (p&lt;0.05). At the same time, the clinical effect was observed earlier in persons in the main group compared to the women in the control group. After treatment, the range of Nugent scores in all women of the main group ranged from 0 to 3 points, in the control group a normal state of biocenosis was diagnosed in 53.3 % of patients, an intermediate state – 20 %, in 26.7 % of women Nugent criteria ranged from 6 to 10 points. In 2 months after the end of therapy, 86.7 % of patients in the main group and 53.3 % of women in the control group had the high effectiveness of the treatment (p&lt;0.05). In 46.7 % of women in the control group, the recurrences of the disease were determined, which required a repeated course of therapy. Only in 13.3 % of patients in the main group, manifestations of atrophic vaginitis were observed. Conclusions. The conducted study proves the high efficiency of suppositories containing hyaluronic acid in the complex treatment of atrophic vaginitis in women of menopausal age, which prolong the inter-recurrence period and reduce the severity of the clinical symptoms of disease recurrences.

Outcomes of Pregnancy Termination of Dead Fetus in Utero in Second Trimester by Misoprostol with Various Regimens.

Objective: To determine the efficacy and adverse outcomes of misoprostol with various regimens for the second-trimester-pregnancy termination of a dead fetus in utero (DFIU). Patients and Methods: A retrospective descriptive study, based on the prospective database, was conducted on pregnancies with dead fetuses in utero in the second trimester. All patients underwent pregnancy termination with various regimens of misoprostol. Results: A total of 199 pregnancies meeting the inclusion criteria were included. The mean age of the participants and the mean gestational age were 30.2 years and 21.1 weeks, respectively. The two most common regimens were 400 mcg injected intravaginally every six hours and 400 mcg taken orally every four hours. In the analysis of the overall efficacy, including all regimens, the mean fetal delivery time was 18.9 h. When considering only the cases involving a delivery within 48 h (success cases), the mean fetal delivery time was 13.6 h. The rates of fetal delivery for all cases at 12, 24, 36, and 48 h were 50.3%, 83.8%, 89.3%, and 93.9%. In the comparison between the various regimens, there were no significant differences in the rate of fetal delivery at 12, 24, 36, and 48 h and adverse effects such as chill, diarrhea, nausea, vomiting, and other parameters such as the requirement for intravenous analgesia, the requirement for curettage for incomplete abortions, the mean total dose of misoprostol, and the rate of postpartum hemorrhage (PPH). Nevertheless, the rate of fever was significantly higher in the regimen of intravaginal insertion of 400 mcg every six hours and that of the requirement for oxytocin was significantly higher in the regimen of oral supplementation of 400 mcg every four hours. Conclusions: The overall success rate within 48 h was 93.6%, which was not different among the various misoprostol regimens. In addition, there were no significant differences in the mean fetal delivery times and the rates of fetal delivery at 12, 24, 36, and 48 h. However, some parameters such as fever, oxytocin requirement, and mean total dose of misoprostol were statistically significant between regimens. In the aspect of global health, misoprostol can be a good option in clinical practice, especially in geographical areas with low-resource levels.

PSI-5 Comparison of 7&amp;7 Synch and 7-day Co-Synch + Cidr Protocols in a Fixed-Time ai System on Beef Cows Using Y-Sorted sex-Selected Semen

Abstract The 7&amp;7 Synch and the 7-day CO-Synch+CIDR fixed-time estrous synchronization protocols were compared among mature and two-year-old lactating postpartum Angus and SimAngus cows (n=201). Mature (n=63) and two-year-old (n=36) cows treated with the 7&amp;7 Synch received an injection of prostaglandin F2α (PGF; 25 mg dinoprost tromethamine) at the same time as insertion of an intravaginal progesterone-releasing insert (CIDR; 1.38g progesterone) on Day -17, gonadotrophin-releasing hormone (GnRH; 100 μg gonadorelin) on Day -10, and PGF at time of CIDR removal on Day -3. Mature (n=65) and two-year-old (n=37) cows in the 7-day CO-Synch+CIDR groups received an injection of GnRH on Day -10 that coincided with CIDR insertion and received PGF at the time of CIDR removal on Day -3. Fixed-timed AI (FTAI) by a single technician using Y-chromosome-bearing sex-sorted semen (4x106 sperm/unit of SexedULTRA 4M) from one of two Charolais bulls occurred on Day 0, 70 hours after PGF injection. Seven days later bulls were pastured with cows for the remainder of the breeding season. Transrectal ultrasound was performed 77-83 days after AI and fetal size was used to differentiate AI and natural service pregnancies. Within the mature cows and two-year-old age groups, the GLIMMIX procedure in SAS was used to compare FTAI and breeding season pregnancy rates with estrous synchronization treatment and sire as main effects. No differences (P=0.41) were observed amongst mature cows in FTAI pregnancy rates between the 7&amp;7 Synch (52.4%) and the 7-day CO-Synch+CIDR (44.9%) treatments. A greater (P=0.05) proportion of two-year-old cows were pregnant to FTAI following treatment with the 7-day CO-Synch+CIDR (56.7%) as compared to the 7&amp;7 Synch (33.4%). A greater (P=0.02) proportion of two-year-old cows in the 7-day CO-Synch+CIDR treatment (91.9%) were pregnant at the conclusion of the breeding season as compared with the 7&amp;7 Synch (69.7%).

Evaluation of luteolysis, follicle size, and time to ovulation in Holstein heifers treated with two different analogs and doses of prostaglandin-F2α

Objectives were to evaluate the effect of 2 analogs of PGF2α (cloprostenol vs. dinoprost) and 2 doses (1 injection vs. 2 injections) on luteolysis, follicle diameter, hormonal concentrations, and time to ovulation in dairy heifers. Holstein heifers were fitted with automated estrus detection devices and had their estrous cycle synchronized using PGF2α and an intravaginal insert containing progesterone. Heifers detected in estrus were blocked by weight and randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement: cloprostenol on d 7 after estrus (CLOx1; n = 45), cloprostenol on d 7 and 8 after estrus (CLOx2; n = 41), dinoprost on d 7 after estrus (DINx1; n = 43), or dinoprost on d 7 and 8 after estrus (DINx2; n = 44). Treatment with the first injection of PGF2α was defined as experiment d 0. Area and blood flow of corpus luteum (CL) and diameter of follicles >5 mm were recorded every 12 h from d 0 to estrus and every 6 h thereafter until ovulation. Blood was sampled every 6 h from d 0 until ovulation. Heifers treated with cloprostenol had shorter interval to luteolysis (± SEM; CLOx1 = 23.5 ± 2.2, CLOx2 = 22.9 ± 2.2, DINx1 = 32.6 ± 2.7, DINx2 = 26.4 ± 2.1 h); however, time to ovulation was not affected by treatment. A smaller proportion of heifers treated with a single injection of PGF2α underwent luteolysis compared with heifers treated with 2 injections (CLOx1 = 84.6 ± 6.2, CLOx2 = 100.0 ± 0.0, DINx1 = 59.7 ± 9.8, DINx2 = 96.3 ± 2.7%). Proportion of heifers that ovulated was smaller for DINx1 compared with other treatments (CLOx1 = 88.8 ± 5.1, CLOx2 = 100.0 ± 0.0, DINx1 = 55.2 ± 9.7, DINx2 = 94.4 ± 3.4%). Ovulatory follicle diameter was larger for DINx1 (18.2 ± 2.7 mm) compared with DINx2 (17.4 ± 2.7 mm), whereas dose did not affect the diameter of the ovulatory follicle in heifers treated with cloprostenol (CLOx1 = 17.6 ± 2.7 vs. CLOx2 = 17.8 ± 2.8 mm). Among heifers that underwent luteolysis, progesterone concentrations from 18 to 36 h after treatment were lesser in heifers treated with cloprostenol compared with those treated with dinoprost. Type of PGF2α did not affect progesterone concentrations past 36 h from treatment; however, heifers treated with 2 PGF2α injections had lesser progesterone concentrations and CL blood flow from 36 to 72 h after treatment compared with heifers that received a single PGF2α injection.

Evaluation of the 7 & 7 Synch and 7-day CO-Synch + CIDR treatment regimens for control of the estrous cycle among beef cows prior to fixed-time artificial insemination with conventional or sex-sorted semen

An experiment was conducted to compare the 7 & 7 Synch and 7-day CO-Synch + controlled internal drug release (CIDR®) treatment regimens before fixed-time artificial insemination (FTAI) of beef cows with conventional or sex-sorted semen. Cows (n = 1538) were blocked based on age and days postpartum (DPP) and randomly assigned to treatment regimen and semen type. Cows assigned to the 7-day CO-Synch + CIDR treatment regimen (n = 769) were administered gonadotropin-releasing hormone (GnRH) and an intravaginal progesterone-releasing insert (CIDR) on Day − 10, and administration of prostaglandin F2α (PG) coincident with CIDR removal on Day − 3. Cows assigned to 7 & 7 Synch (n = 769) were administered PG and a CIDR device on Day − 17, GnRH on Day − 10, and PG coincident with CIDR removal on Day − 3. Cows were administered GnRH coincident with FTAI, which was performed 66 h after CIDR removal with conventional (20 × 106 cells) or sex-sorted (4 × 106 cells) semen. Expression of estrus was affected by treatment regimen (P = 0.01) and by treatment regimen × DPP (P = 0.0004), as a result of imposing the 7 & 7 Synch regimen; therefore, a greater percentage of cows expressed estrus (82% compared with 64%), particularly among cows with greater DPP. Pregnancy percentages resulting from FTAI were less (P < 0.0001) when using sex-sorted semen but greater among cows treated with 7 & 7 Synch (conventional semen: 72%; sex-sorted semen: 52%) compared with 7-day CO-Synch + CIDR (conventional semen: 61%; sex-sorted semen: 44%).