PGF2α Analogue Research Articles

Vaginal cytology during the estrous cycle and early pregnancy of ewes

Changes in vaginal epithelial cells and blood progesterone and estrogen concentrations were compared between estrous cycle and early pregnancy in multiparous Sanjabee ewes. Twenty non-pregnant ewes were synchronized with intravaginal insertion of a controlled internal drug release (CIDR) device which was in place for 7 days, intramuscular administration of a GnRH analog (Alarelin acetate; 12.5 µg) at the time of CIDR insertion (Day -7), a PGF2α analog (D-cloprostenol sodium; 125 µg) and 500 I.U. of human chorionic gonadotropin (hCG) at the time of CIDR removal (Day 0). At Day 0, all ewes were introduced to four fertile rams and observed for estrous signs. Ewes exhibiting estrous signs were divided into two groups: MATED (confirmed pregnant with transrectal ultrasound scanning at Day 35) and NOT-MATED (confirmed non-pregnant with transrectal ultrasound scanning at Day 35). Vaginal smears and blood samples were taken daily and every other day, respectively, from the beginning of estrus for 20 days for cytology and hormone assay. Based on the results, during days 0 and 1 of the estrous cycle, only the percentages of intermediate cells differed between the groups (P < 0.05). During days 2 to 4, there was no difference in the cell populations between the groups (P ≥ 0.05). During days 5 to 16, the percentages of all cell types except the parabasal cells differed between the groups (P < 0.05). During days 17 to 20, the percentages of all types of vaginal epithelial cells differed between the groups (P < 0.05). Progesterone concentrations increased gradually from days 0 to 14 of the estrous cycle in both groups, however, they decreased significantly afterward in the NOT-MATED group. Estrogen concentrations changes showed an opposite pattern to that of progesterone in the study groups. Collectively, vaginal cytology can be used as a useful tool in assessing hormonal and physiological characteristics of the female reproductive system and thus provides a more accurate understanding of the physiology of estrous cycle and early pregnancy in ewes, which can be used to improve reproductive management.

Comparative study between intralesional injection of prostaglandin F2α alone or combined with corticosteroids in the treatment of alopecia areata

Background Alopecia areata (AA) is an autoimmune disorder causing hair loss, yet its treatment remains challenging due to limited Food and Drug Administration approved therapies. Prostaglandin F2α (PGF2α) has emerged as a potential therapy. Objective To assess the efficacy of PGF2α intralesional injection in AA treatment. Patients and methods This case comparative work was performed on 75 localized AA patients who were split into three groups receiving different intralesional injections: group A received triamcinolone acetonide (TRA), group B received a combination of TRA and latanoprost (a PGF2α analog), and group C received latanoprost alone. Treatment response was assessed using the Severity of Alopecia Tool score, Severity of Alopecia Tool II, and Alopecia Areata Severity Index. Results High treatment response was observed in 76% of group B, compared to 64% in group A and 40% in group C. Group B showed the most significant reduction in severity score (median=0), followed by group A (median=1), and group C (median=3). Multivariate analysis identified group B as a significant predictor of a high treatment response (odds ratio=10.292, 95% confidence interval=1.904–55.632, P=0.007). Conclusion Intralesional injection of TRA and latanoprost (PGF2α analog) demonstrated superior efficacy in treating localized AA compared to other treatment modalities.

Hair Growth Promoting Effects of 15-Hydroxyprostaglandin Dehydrogenase Inhibitor in Human Follicle Dermal Papilla Cells.

Prostaglandin E2 (PGE2) is known to be effective in regenerating tissues, and bimatoprost, an analog of PGF2α, has been approved by the FDA as an eyelash growth promoter and has been proven effective in human hair follicles. Thus, to enhance PGE2 levels while improving hair loss, we found dihydroisoquinolinone piperidinylcarboxy pyrazolopyridine (DPP), an inhibitor of 15-hydroxyprostaglandin dehydrogenase (15-PGDH), using DeepZema®, an AI-based drug development program. Here, we investigated whether DPP improved hair loss in human follicle dermal papilla cells (HFDPCs) damaged by dihydrotestosterone (DHT), which causes hair loss. We found that DPP enhanced wound healing and the expression level of alkaline phosphatase in DHT-damaged HFDPCs. We observed that DPP significantly down-regulated the generation of reactive oxygen species caused by DHT. DPP recovered the mitochondrial membrane potential in DHT-damaged HFDPCs. We demonstrated that DPP significantly increased the phosphorylation levels of the AKT/ERK and activated Wnt signaling pathways in DHT-damaged HFDPCs. We also revealed that DPP significantly enhanced the size of the three-dimensional spheroid in DHT-damaged HFDPCs and increased hair growth in ex vivo human hair follicle organ culture. These data suggest that DPP exhibits beneficial effects on DHT-damaged HFDPCs and can be utilized as a promising agent for improving hair loss.

Pharmacotherapy of glaucoma in terms of evidence-based medicine

Glaucoma is a disease associated with increased intraocular pressure (IOP). Of the pharmacological agents for treating glaucoma, there are drugs of the first (most effective and safe) and second-line treatment. First-line treatment includes prostaglandin analogs and beta-blockers. The currently used prostaglandin analogs (latanoprost, bimatoprost, tafluprost and travoprost) are PG F2α analogs that act through stimulation of FP receptors. They are distinguished by the optimal ratio of effectiveness and risk of side effects. They are convenient for the patient because for the therapeutic effect, it is enough to prescribe 1 time per day. As a result, it is rational to start the treatment of glaucoma with a drug in this group. In terms of pharmacoeconomics, the most affordable prostaglandin drug is latanoprost, which is generally as effective as other prostaglandin analogs. β-adrenergic blockers reduce the production of intraocular fluid, the formation of which is controlled by β1- and β2-adrenergic receptors. Therefore, non-selective β-blockers (timolol, levobunolol, metipranolol, and carteolol) have a pharmacodynamic advantage over selective β1-adrenergic antagonists (betaxolol). Conducted clinical studies of β-blockers have shown that given the cost, efficacy and safety, timolol was the most preferable treatment for glaucoma. In the presence of medical contraindications to the use of first-line drugs or to enhance their effectiveness, α2-agonists (apraclonidine and brimonidine), carbonic anhydrase inhibitors (usually local action: dorzolamide and brinzolamide), M-cholinomimetics (pilocarpine, carbachol and echothiopate), and also Rho-kinase inhibitors (ripasudil)

Effect of cloprostenol on luteolysis in jennies

An intensive farm system has progressively emerged with the rapid development of the donkey industry. One of the primary challenges impeding the development of the donkey industry and the preservation of endangered donkey species is the low reproductive efficiency of jennies (Feng Yang et.al. Equine Veterinary Journal 2021, 53: 1218-1226). In donkeys, luteolysis occurs between days 15 and 17 after ovulation (Carluccio A. et.al. Theriogenology, 2007, 68(6): 914-919). To shorten the estrus cycle, a new estrus cycle may start after luteolysis is induced by the PGF2α analog (Duccio P. et.al. Theriogenology, 2018, 121: 175-180). Treatment of mares and cattle with PGF2α analogs from the fifth day after ovulation induces luteolysis (Bergfelt DR. et.al. Theriogenology, 2006, 65(8): 1605-1619). However, information regarding luteolysis in jennies is limited. In this study, we aimed to investigate the effect of cloprostenol on luteolysis in jennies. The PGF2α analog used in the experiment was cloprostenol (0.2 mg/2 ml, Ningbo Second Hormone Factory, Zhejiang, China). To study the effects of different cloprostenol doses on the estrus cycle, jennies were treated with 0.2 mg (n=30), 0.4 mg (n=30), or 0.6 mg (n=30)cloprostenol (PGF2α), respectively. The time of luteolysis, ovulation interval, follicular size at ovulation, and ovulation rates were compared between the groups. The results showed that luteolysis rates in the jennies within 72 h of treatment were higher than 90% in the three treatment groups. The time required for luteolysis was not statistically different among the three groups. In addition, the estrus interval, ovulation interval, ovulation diameter, and ovulation rate did not differ among the three groups. To examine the timing effect of cloprostenol treatment on the estrus cycle, jennies were treated with 0.4 mg cloprostenol on the third (D 3, n=30), fifth (D 5, n=30), or seventh (D 7, n=30) day following ovulation, respectively. The results showed that the luteal phase of the D 3 treatment group was shorter than that of the D 5 and D 7 treatment groups, and in the D 5 treatment group was shorter than that of the D 7 group (P < 0.05). In can be concluded that treatment with 0.4 mg cloprostenol on the third day following ovulation shortens the length of the luteal phase and estrus cycle considerably.

Prostaglandin F2α regulates mitochondrial dynamics and mitophagy in the bovine corpus luteum.

Prostaglandins are arachidonic acid-derived lipid mediators involved in numerous physiological and pathological processes. PGF2α analogues are therapeutically used for regulating mammalian reproductive cycles and blood pressure, inducing term labor, and treating ocular disorders. PGF2α exerts effects via activation of calcium and PKC signaling, however, little is known about the cellular events imposed by PGF2α signaling. Here, we explored the early effects of PGF2α on mitochondrial dynamics and mitophagy in the bovine corpus luteum employing relevant and well characterized in vivo and in vitro approaches. We identified PKC/ERK and AMPK as critical protein kinases essential for activation of mitochondrial fission proteins, DRP1 and MFF. Furthermore, we report that PGF2α elicits increased intracellular reactive oxygen species and promotes receptor-mediated activation of PINK-Parkin mitophagy. These findings place the mitochondrium as a novel target in response to luteolytic mediator, PGF2α. Understanding intracellular processes occurring during early luteolysis may serve as a target for improving fertility.

Application of the "echo-synch protocol" to advance pregnancy onset in ewe lambs at the first reproductive season.

This study assessed the efficacy and economic impact of a reproductive protocol based on repeated ultrasound scanning (US) associated with the use of GnRH to advance pregnancy onset in ewe lambs. Prepubertal ewe lambs (n = 133) were divided into three weight groups (High: HW n = 35; Medium: MW n = 65; Low: LW n = 33). Thereafter, animals were randomly allocated into two subgroups: GnRH, ewe lambs treated with GnRH analog and then exposed to rams; CTR, ewe lambs exposed to rams only. CTR groups were joined with rams as a single flock. GnRH groups were kept separate from rams receiving a single dose of gonadorelin (40 μg/head) and then were evaluated after a week of US. Animals showing corpora lutea received an injection of PGF2α analog (100 μg/head) and then were joined with rams. The remaining ewe lambs received a second dose of gonadorelin and were kept separate from the rams. After another week, animals were checked again and the ones showing corpora lutea were injected with the PGF2α analog, while the others received a third injection of gonadorelin. On the same day, all the animals were joined with rams. Pregnancies were confirmed within 30 days by US. The efficacy of the protocol was determined by assessing differences in the number of days required to achieve pregnancy rates of 25, 50, and 75% and in the total costs and incomes from birth to the end of first lactation within the groups. The GnRH-MW group showed the best performances in reaching the threshold pregnancy rates of 25, 50, and 75%, but the effect of treatment was significant only at the 25% threshold (p < 0.01). Both low groups displayed an overall poorer performance at 50 and 75% thresholds than medium and high-weight groups (p = 0.01 and p < 0.01, respectively). The GnRH administration did not advance pregnancy onset in GnRH-HW compared with CTR-HW. In the balance between costs and income, the HW-CTR and MW-GnRH groups showed higher gross margins than the other groups. Using the US/GnRH protocol in ewe lambs appears technically and economically effective in animals that have not reached the optimal weight at the first breeding season, advancing ewe lambs' pregnncies and increasing farm profitability.

The efficacy of bimatoprost ophthalmic solution combined with NB-UVB phototherapy in non-segmental and segmental vitiligo: a single-blind randomized controlled study

Bimatoprost ophthalmic solution 0.03% (PGF2α analogues) combined with narrowband ultraviolet B (NB-UVB) was reported to be an effective treatment for vitiligo. To investigate the efficacy and safety of treatment for non-segmental/segmental vitiligo compared among bimatoprost ophthalmic solution 0.01% combined with NB-UVB phototherapy, bimatoprost monotherapy, and placebo. This single-blind randomized controlled study enrolled stable Thai vitiligo patients with at least three similarly sized lesions in the same anatomical area. The treatment duration was 6 months with 1- and 2-month post-treatment follow-ups. The 3 selected lesions on each patient were randomized to receive combination therapy, monotherapy, or placebo. The Vitiligo Area Scoring Index (VASI) was used to evaluate lesion response. Of the 25 initially enrolled subjects, 19 patients were analyzed. There were 13 and 6 non-segmental and segmental vitiligo cases, respectively. Eight and 11 cases had face/neck and non-face/neck lesions, respectively. Non-segmental vitiligo and non-face/neck vitiligo patients in the combination group had significant improvement in VASI score at 3 months, 6 months, and at the 2-month follow-up. No side effects were observed/reported. Bimatoprost combination therapy was shown to be safe and effective for treating Thai patients with non-segmental vitiligo in non-face/neck areas of the body.

Effect of Age and Different Doses of GnRH and PGF2α Analogue on Oesturs Synchronization and Conception Rate of Crossbreed Heifers

Forty-eight crossbreed heifers were artificially inseminated within 10-20 hours after standing heat to evaluate the effects of different doses of hormones and age on reproductive performance synchronized with GnRH and PGF2α. On day 90 after artificial insemination, the conception rate was confirmed by rectal palpation. There were significant differences (<I>P</I><0.05) in oesturs response in maximum doses (1.5 ml GnRH used in group III) of the hormone-treated group. The conception rate was significantly higher (<I>P</I><0.05) in the highest treatment group (1.5 ml, 66.67%) than in low treatment groups (1 ml and 1.3 ml; 42.85% and 44.44%; respectively) in morning synchronization protocol (83.33%). The effect of different doses of the hormone on conception rates was statistical significance on the age of crossbreed heifers (<I>P</I><0.05), time of initiation of ovsynch (<I>P</I><0.05), time of insemination (<I>P</I><0.05), semen placement (<I>P</I><0.05), body condition score (<I>P</I><0.05), retrieve to natural oesturs and subsequent pregnancy rate (<I>P</I><0.05). On the other hand, there was no significant (<I>P</I>>0.05) effect on conception rate by the onset of oesturs and duration of oesturs. The highest oesturs rate and conception rate was observed at 75.00% and 66.67%; respectively by administration of maximum doses of the hormone of body condition score (3) 69.23% of crossbreed heifer. When 1.5 ml GnRH was used in group III, the conception rate was observed higher at 25-30 months of age (77.78%) which were inseminated in the site of the deep cervix (77.77%) with frozen-thaw semen by experience inseminator at 10-14 hours (75%) after the onset of oesturs and naturally retrieve of oesturs on conception rate (75%). It was concluded that the age of the heifers, doses of the hormone, Body Condition Score (BCS), time of initiation of synchronization protocol, time of AI, semen placement and retrieve to oesturs naturally were the most important factors to get maximum oesturs response and conception rate of crossbreed heifers. So, it may be suggested that to achieve the desired rate of oesturs and conception, the farmers should inseminate their crossbreed heifers (BCS, 3 out of 5) with maximum doses of hormone in the deep cervix by technically sound and skilled inseminator at 10-14 hours after onset of oesturs in morning synchronization protocol. GnRH and PGF2α analogs could successfully induce oesturs response and improve the fertility of a dairy herd. Therefore, ovsynch protocol for crossbreed heifers is needed for further work involving a large number of animals for a definite conclusion.

SUPEROVULATORY RESPONSE IN DAIRY COWS AFTER APPLICATION OF ECG

PURPOSE: The aim of the present study was to determine the follicular dynamics and superovulatory response in dairy cows after the administration of eCG. METHODS: The study included 6 Holstein cows bearing CL in the ovaries treated with PGF2α for estrus induction. After estrus detection daily ultrasound scanning to monitor the dominant follicle of the first follicular wave was made. Superovulation treatment (2500 IU eCG and two doses of 500μg PGF2α analog on the 48th and 60th hour after the eCG injection) started at the emergence of the second follicular wave. The number and diameter of the follicles ≥ 5 mm, the number of corpora lutea and anovulatory follicles (AF) and the ovulation rate (OR) were estimated. RESULTS: At the start of the superovulation treatment (SO) the average number and diameter of the medium and large follicles were 2.7±1.2 and 0.67±0.13 cm and 1.6±0.5 and 1.45±0.28 cm, respectively. There were high positive correlations between the days of the treatment and number of large and medium follicles (r = 0.80, P = 0.018 and r = 0.75, P = 0.03). The average number of large follicles on day 5 was greater than those at the start of SO. On day 8 the OR was 55.9% and the difference between anovulatory follicles and corpora lutea was not significant. CONCLUSIONS: The used protocol provided acceptable OR, but the average number of AF (44.1%) remained high. Ultrasonography could be used to monitor the first dominant follicle and the emergence of a new follicular wave.

Low plasma progesterone concentration during the early luteal phase delays endometrial development and the beginning of placentation in mares

While detrimental effects of reduced plasma progesterone concentration in the early luteal phase on conceptus development in horses have recently been demonstrated, there is no information on associated effects on the endometrium, allantochorion (AC), and chorionic girdle (CG) in this species. We hypothesised that reduced early postovulatory progesterone concentration in pregnant horses is detrimental to endometrial function and development of the embryonic membranes and is an underlying cause of delayed conceptus development. After insemination and ovulation, mares (n = 11) were assigned to treatment (TREAT) or control (CON) during two pregnancies. In TREAT pregnancies, mares received a PGF2α analogue for four consecutive days starting on the day of ovulation with the aim to reduce progesterone secretion. Mares were left untreated in CON pregnancies and thus served as their own controls. Endometrial biopsies for analysis of histomorphology, epidermal growth factor (EGF) and EGF receptor (EGFR) mRNA and protein expression in the endometrium, AC, and CG as well as abundance of regulatory T lymphocytes (Tregs) were collected on day 34 of pregnancy. Histomorphometric analysis revealed a higher luminal endometrium and a higher CG epithelium in CON compared to TREAT pregnancies. Abundance of mRNA for EGF and EGFR was large in the endometrium, AC and CG but did not differ between TREAT and CON pregnancies. The number of endometrial regulatory T lymphocytes was reduced in TREAT compared to CON pregnancies, adding further aspects to the potentially detrimental effects of reduced progesterone concentrations on equine pregnancy.

The vaginal and uterine blood flow changes during the ovsynch program and its impact on the pregnancy rates in Holstein dairy cows

AimOvSynch is a hormonal protocol for synchronization of estrus and use of artificial insemination (AI) at an optimal time without adverse effects on the ovaries or uterus. This study investigated the use of noninvasive color Doppler ultrasound to assess changes in uterine and vaginal blood flow during the Ovsynch program for synchronization of estrus and its relation to the pregnancy rates in Holstein cows.Materials and methodsThe experimental cows received an intramuscular dose of 10 μg of a GnRH analogue (G1), followed 7 days later with an intramuscular injection of synthetic prostaglandin F2α (P: PGF2α) analogue (500 μg cloprostenol sodium), and given a 10 μg, injection of the GnRH analogue (G2) i.m. 48 h after the PGF2α treatment, and the cows were bred 14-16 h after. Uterine and vaginal perfusion were investigated by performing transrectal Doppler ultrasonography of both the uterine and vaginal arteries in Holstein cows at different time points during the Ovsynch program to determine: peak systolic velocity (PSV), time-averaged maximum velocity (TAMV), the volume of blood flow (BFV), pulsatility index (PI), resistance index (RI), resistance impedance (S/D) and diameters of uterine (UA) and vaginal (VA) arteries. Steroid hormones were also assayed. Transrectal ultrasonography (TUS) was performed at 32 and 60 days to confirm the pregnancy per artificial insemination (P/AI).ResultsThe uterine PSV, TAMV, and PV were greater at the time of the cloprostenol sodium and second GnRH injections (p<0.05) than at the time of the first GnRH injection. The vaginal PSV, PV were greater at the time of the cloprostenol sodium than at the time of the first and second GnRH injections (p<0.05). The receiver operating characteristic curve (ROC curve) indicated a high correlation between the uterine and vaginal blood flow and the rate of the pregnancy (p<0.05). The area under the ROC curve was 0.920 and 0.87 (p<0.05) for vaginal and uterine arteries respectively at time of G2. The serum levels of progesterone, estrogen and cortisol were correlated with the P/AI (p<0.05). The P/AI significantly decreased from 43.9 % at 32 d to 35.37 % at 60 d.ConclusionThese results indicate that noninvasive Doppler ultrasonography is a valid method to evaluate changes in the characteristics of uterine and vaginal blood flow in cows during the Ovsynch protocol. Furthermore, vaginal and uterine blood flow are two determinant factors for the higher conception rates in Holstein dairy cows.

Comparing the effect of estradiol benzoate and 17β-estradiol plus progesterone on follicular turnover and development, and pregnancy outcomes in a timed artificial insemination protocol

The objective of this study was to compare the efficiency of estradiol benzoate (EB) and 17β-estradiol (E2) associated with progesterone (P4) in a fixed-time artificial insemination (FTAI) protocol. We hypothesized that E2+P4 induces an earlier emergence of a new follicular wave (NFW), improving pre-ovulatory follicle diameter and pregnancy rates to FTAI (P/FTAI). In Exp.1, on Day 0 (D0), all Bos indicus cows (n = 12/group) received an intravaginal P4 device and a dose of PGF2α analogue. On D0, females were randomly assigned to receive EB or E2+P4. On D8.5, P4 intravaginal devices were removed and a dose of PGF2α and EB were administered in all females followed by fixed-timed AI on D10. Between D0 and D10, the dominant follicular growth was determined by ovary ultrasonography exams. On D8.5 and D10 the percentage of color power-Doppler signals in the dominant follicular wall was evaluated. In Exp. 2, 467 females (2-year-old nulliparous [n = 76], primiparous [n = 92] and pluriparous [n = 299]) were subjected to the similar FTAI and assigned to be treated with EB (n = 243) or E2+P4 (n = 224). Pregnancy diagnosis was performed 30 days after FTAI by ultrasonography. The day to emergence of NFW was similar between treatments (EB: 3.7 ± 0.37 vs. E2+P4: 3.3 ± 0.3, P = 0.76). Females treated with E2+P4 presented greater (P = 0.06) follicular growth between the emergence and D9 (1.18 ± 0.07) than those treated with EB (0.97 ± 0.08). There was also a positive effect (P < 0.05) of E2+P4 on diameter of the dominant follicle on D9 (13.0 ± 0.6 vs. 10.9 ± 0.55) and blood perfusion of the follicle wall on D8.5 (49 vs. 40%). There was a treatment by parity category interaction effect on P/FTAI (P < 0.05). Treatment with E2+P4 was advantageous to P/FTAI of primiparous cows (E2+P4: 58% and EB: 30%). However, for nulliparous and pluriparous cows, P/FTAI was similar between treatments (∼50%). In conclusion, in a E2/P4-based protocol for FTAI, E2+P4 is as efficient as EB in inducing new follicular emergence within a similar day range, but it results similar or greater P/FTAI.

Screening and quantification of undeclared PGF2α analogs in eyelash-enhancing cosmetic serums using LC-MS/MS

In recent years, cosmetics deemed equivalent to pharmaceutical products containing prostaglandin F2α (PGF2α) analogs have been distributed overseas in the form of eyelash serums that can be purchased via the internet. The purpose of this study was to investigate the presence or absence of PGF2α analogs in eyelash serums procured in Japan via the internet to elucidate the actual composition. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) measurement system was developed for the determination of 14 PGF2α analogs in cosmetic serums. In total, 64 eyelash serum samples were purchased from 34 websites. After pretreatment, eyelash serum samples were screened for PGF2α analogs using the LC-MS/MS system. Products containing PGF2α analogs were subjected to quantification of these compounds. Of the 64 products, four were found to contain bimatoprost, among which, three did not indicate their contents on their package labels. In contrast, no samples were found to contain latanoprost, travoprost, or tafluprost, which are prescribed for glaucoma treatment. Additionally, eight products contained other PGF2α analogs, which have not been used as pharmaceuticals. The ease of access to cosmetic serums containing PGF2α analogs via online purchases presents a risk of serious side effects, particularly when consumers are not informed of their contents on the packages. This issue requires serious consideration to avoid the incorporation of pharmaceutical substances into cosmetic products.

Increased AMH plasma concentration and small antral follicles - Deslorelin slow release implants as a promising tool for preparation of mares for OPU?

A major factor affecting oocyte yield during ovum pick-up (OPU) procedures in mares is the number of follicles present on the ovary and the associated number of recovered oocytes (Cuervo-Arango et al. Reproduction, Fertility and Development. 2019; 31(12): 1894-1903). The interest in manipulation of ovarian follicular populations in horses is increasing because of benefits with regard to the oocyte yield and the overall outcome for assisted reproductive procedures. This study aimed to evaluate effects of slow-release implants containing deslorelin acetate on antral follicle counts (AFC), follicular subpopulations of different size ranges and plasma anti-Mullerian hormone (AMH) and gonadotrophin concentration in mares. Haflinger mares (n=12) were synchronized with two injections of a PGF2α analog. One day after the second injection, mares were randomly assigned to the slow-release implant (DES, 9.4mg deslorelin implant, n=6) or the control group (CON, deslorelin short-acting formulation, 1.25mg, n=6). Transrectal examination of the genital tract and collection of blood samples for the analysis of progesterone, AMH and gonadotrophins were performed until day 90 and on days of AFC assessment. AFC and follicle numbers according to size groups were determined on days 0, 10, 25, 50, 90, 180, 270 and 360 after implant insertion. The interval from implant insertion to the first spontaneous ovulation was prolonged in group DES (23.8±10.5 days) compared to CON(17.0±3.9 days) (p<0.05). Treatment influenced concentrations of LH and FSH over time (p<0.05) with the concentration of LH being greater in DES than in CON mares on days 30 and 55 and of FSH on day 15 after implant insertion, respectively (p<0.05). AMH concentration was greater in group DES from day 30 onwards (interaction treatment x time p<0.05). The AFC and the number of follicles with 5 to 10, 10 to 15 and 15 to 20 mm in diameter changed over time (p<0.05). A time x treatment interaction was demonstrated for follicles of 10 to 15 mm in diameter with more follicles of this size present in DES than in CON mares on day 90 (p<0.05). In group DES, concentration of AMH and the AFC (r=0.623, p<0.001) as well as concentration of AMH and the number of follicles 5 to 20 mm in diameter (r=0.442, p<0.01) were positively correlated whereas there was no such correlation in group CON. In conclusion, long-acting deslorelin implants induce changes in follicular development reflected in AMH plasma concentration. These changes are present from day 30 after implant insertion, suggesting that not only direct effects of deslorelin on gonadotrophin secretion but also the reduction of FSH suppressing factors from follicles such as inhibin may be involved. Treatment of mares with 9.4mg deslorelin slow-release implants might thus be a promising tool as preparation for OPU.

Effect of Echinops spinosus (Teskra) and PGF2α on reproductive performance in cows with clinical endometritis

The efficacy of Echinops spinosus (Teskra) and PGF2α in the treatment of clinical endometritis (CE) on clinical recovery and reproductive performance was evaluated. A total of 20 Holstein-Friesian cows with CE that were 21-40 days in milk (DIM) were assigned to one of two experimental groups. Cows in the PGF group (n=10) received two doses of d-cloprostenol (analogue of PGF2α) 14 days apart. Cows in the Teskra group (n=10) received a decoction of Teskra root administered orally once daily for three days. The control group consisted of 20 clinically healthy cows (n=20; HE group). The clinical recovery rate was 70% and 40% in PGF and Teskra groups, respectively (P&amp;gt;0.05). The proportion of pregnant cows at first service was 35%, 40%, 20% in the HE, PGF, and Teskra groups, respectively (P&amp;gt;0.05). However, the conception rate until 120 DIM was significantly higher in the HE group compared to the Teskra group (P&amp;lt;0.05). The mean days from calving to the first service interval (CFSI) were significantly lower in the HE group compared to the PGF and Teskra groups (P&amp;lt;0.001). Cows in the Teskra group conceived approximately 35 d and 26 d later than cows in the HE and PGF groups, respectively (P&amp;lt;0.05). Cows in the Teskra group required about 2.4 services to become pregnant, compared to 1.8 and 1.6 for those in the HE and PGF groups, respectively (P&amp;gt;0.05). Except for CFSI, no significant differences were found between the HE and PGF groups for any of the tested parameters. The results indicate that endometritic cows treated with PGF2α had nearly the same reproductive performance as healthy cows and Teskra was not effective in treating CE in dairy cows.

Fertility Response Following Estradoublesynch and Progesterone Based Ovsynch Protocols in Delayed Pubertal Buffalo Heifers

The study evaluated the fertility response following Estradoublesynch and progesterone-based Ovsynch protocols in delayed pubertal buffalo heifers. A total of twenty delayed pubertal buffalo heifers were subdivided in two equal subgroups. Group I animals were treated with PGF2α analogue (500 µg cloprostenol sodium) i.m. on day 0, GnRH analogue (20 µg buserelin acetate) on day 2, PGF2α analogue (500 µg cloprostenol sodium) repeated on day 9 and estradiol benzoate 1 mg intramuscular (i.m.) on day 10. Fixed-time artificial insemination (FTAI) was done at 48 and 60h. Group II animals were subjected to GnRH analogue (20 µg buserelin acetate) i.m. along with CIDR insert intravaginally on day 0, PGF2α analogue (500 µg cloprostenol sodium) i.m. and removal of CIDR on day 7, GnRH analogue on day 9 followed by FTAI at 12 and 24h. Blood collection (plasma P4) and ultrasonography (luteal and follicular dynamics) was performed on day 0, 9, 10, 12 in group I and day 0, 7, 9 and 10 in group II animals. Pregnancy diagnosis was made at 45-60 days post AI. Estrus response was observed in all the buffalo heifers of group I and II. First service conception rate was significantly higher in group I (50%) compared to group II (20%) (P&lt;0.05). Overall pregnancy rate was almost similar in both group I and II (60% vs 70%). In conclusion, estradoublesynch and progesterone-based ovsynch protocol can effectively induce ovulatory estrus and improve pregnancy rates in delayed pubertal buffalo heifers.

Application of PGF2α at the moment of fixed-time artificial insemination in crossbred beef cows.

Although studies have shown positive effects of gonadotropin releasing hormone (GnRH) or prostaglandin F2α (PGF2α) at the moment of fixed-time artificial insemination (FTAI) in the conception rate (CR) of cattle, its effects on treatments based on progesterone (P4) and estradiol benzoate (EB) is still not conclusive. The objective of this study was (1) to evaluate the effect of a PGF2α analogue at FTAI in the CR of crossbred beef cows submitted to a 11d FTAI protocol based on P4 and EB; and (2) to describe the CR between PGF2α-treated and control cows in different body condition scores (BCS) and parity categories. Crossbred (½ Nellore and ½ Angus) beef cows were submitted to a synchronization protocol and randomly assigned into 2 groups: Control (n = 163), at FTAI cows received 2 mL of saline solution as a placebo, and PGF2α (n = 163), at FTAI cows were treated with PGF2α analogue (10 mg of dinoprost tromethamine). Pregnancy diagnosis was performed 33d post-FTAI. Binary logistic regression model was used to analyze the effect of PGF2α treatment on CR. There was no difference in CR between PGF2α and control groups (P > 0.05; odds ratio (OR) = 0.92; confidence interval (CI) = 0.59-1.4). A greater CR was found in heifers (P = 0.0006, OR = 2.65, CI = 1.61 - 4.38) and multiparous (P = 0.0006, OR = 2.12, CI = 1.04 - 4.3) when compared to primiparous cows. Cows with low BCS (4; 9-point scale) showed lower CR when compared with moderate BCS (5-6; 9-point scale) (P < 0.05; OR = 0.10; CI = 0.06 - 0.18). There was no numerical difference on CR between PGF2α-treated and control cows in different BCS and parity categories. The results suggested that the CR in this study was not influenced by 10 mg PGF2α analogue at FTAI.

Cefacetrile and Rifaximin association might improve first service conception rate and reduce the number of services per conception in cows with clinical endometritis

Clinical endometritis (CE) is a serious disease leading to poor reproductive performances in lactating dairy cows, thus diminishing farm profitability. To preserve optimum reproductive efficiency, various strategies and therapeutic approaches have been proposed to manage cows with CE, often with contradictory results. Thus, investigating new paths to CE treatment is economically important. The aim of the present study was to test the efficacy of three therapeutic protocols on the clinical cure rate of CE, and improvement of reproductive performance. Cows with CE (n=42), 21–38 days in milk (DIM), were assigned to three treatment groups: PGF: cows (n=19) were treated systemically with two doses of d-cloprostenol, a PGF2α analogue, at 14-days intervals; CEFAX: cows (n=10) received an intrauterine infusion of the combined antibiotics Cefacetrile and Rifaximin; and NAX: cows (n=13) received systemic treatment with Ceftiofur crystalline free acid (CCFA). A control group included cows (n=36) free from CE (healthy group: HE). All cows were clinically re-examined after the end of the treatment protocol. The clinical cure rate was 73.7%, 80% and 69.2% in PGF, CEFAX, and NAX groups, respectively (P&amp;gt;0.05). The HE group had a significantly shorter calving to first service interval compared to CEFAX and PGF groups (P&amp;lt;0.05), however the difference was not significant with NAX group. The mean calving to fertilizing service interval (CFI) was slightly higher in all three treatment groups compared to the HE group, however the difference was not significant (P&amp;gt;0.05). CEFAX protocol resulted in shorter but not statistically significant CFI, compared to the PGF and NAX protocols. Services per conception rate were slightly lower (1.7) in the CEFAX group compared to HE (1.75), PGF (1.84) and NAX (2.23) groups, however these differences were not significant. First service conception rate and conception rate at 105 DIM did not differ statistically between the treatment groups. While the difference was not significant, CEFAX protocol had slightly better cure rate for CE, reducing the number of services per conception and boosting the resumption of ovarian activity after calving. Validating these finding on a larger herd size will improve the accuracy of these findings.

PSV-4 Peri-estrus ovarian, uterine, and hormonal variables influence the uterine luminal fluid metabolome in beef heifers

Abstract In cattle, uterine luminal fluid (ULF) is the main source of molecules that support embryo development and survival during the peri-implantation period. Overarching hypothesis was that peri-estrus changes in ULF volume through accumulation and resorption mechanisms influence ULF composition during the estrous cycle and early pregnancy. Objectives were (1) to characterize individual temporal and spatial changes in ULF volume, endometrial and luteal vascularity, endometrial and luteal size, and progesterone (P4) concentrations during the peri-estrus period in beef heifers and, (2) associate such changes with the metabolite composition in the ULF, four days after estrus. Fourteen Bos indicus heifers that presented a PGF2α responsive CL received 500 µg PGF2α analog i.m. and were examined daily by rectal B-mode and pulse-wave color-Doppler ultrasonography until the fifth day after estrus (estrus = d 0). Plasma P4 was measured daily. On d 4, the uterine body was sampled using a cytology brush for targeted metabolomic analysis by mass spectrometry. Multivariate analyses clustered heifers according to ovarian, uterine, and hormonal variables in clusters A (n = 5) and B (n = 8 heifers). Individual metabolite concentrations were compared between clusters A and B by univariate analysis using t-test after FDR correction. Concentrations of Pro, Ala, Leu, Gly, Val, Lys, Ile, Phe, Asp, Orn, Tyr, Arg, Trp, Suc, Cit, ADMA, the sum of essential Amino Acids (AA), sum of non-essential AA, sum of aromatic AA, and total AA were greater in cluster A (FDR ≤ 0.05). ULF volume dynamics and associated uterine, ovarian, and hormonal variables during the peri-estrus period presented a concerted variation among heifers, which was associated with the ULF composition four days after estrus. Potential implications for embryo receptivity and reproductive outcomes are the focus of the current investigation.