Serum P4 Concentrations Research Articles

The effect of rifampin on serum etonogestrel concentrations and biomarkers of ovulation among contraceptive implant users: A pharmacokinetic and pharmacodynamic study

ObjectivesRifampin, a strong CYP3A inducer, is the gold standard for evaluating CYP3A-mediated drug-drug interactions. We aimed to evaluate the pharmacokinetic and pharmacodynamic effects of a short course (2 weeks) of rifampin on serum etonogestrel (ENG) concentrations and serologic measures of ovarian activity (endogenous estradiol [E2] and progesterone [P4]) among ENG implant users. Study designWe enrolled healthy females using ENG implants for 12 to 36 months. We measured baseline serum ENG concentrations using a validated liquid chromatography mass-spectrometry assay and baseline E2 and P4 concentrations using chemiluminescent immunoassays. After 2 weeks of rifampin 600 mg daily, we repeated ENG, E2, and P4 measurements. We compared pre- and post-rifampin serum measurements using paired Wilcoxon signed-rank tests. ResultsFifteen participants completed all study procedures. Participants’ median age was 28.2 years (range 21.8–34.1), median body-mass index was 25.2 kg/m2 (range 18.9–37.3), and median duration of implant use was 22 months (range 12–32). All participants experienced significant decreases from baseline ENG concentrations (median 164.0 pg/mL [range 94.4–265.0]) to post-rifampin measurements (median 47.8 pg/mL [range 24.7–82.8]) (p < 0.001). Serum E2 concentrations also significantly increased with rifampin exposure (median 73 pg/mL vs 202 pg/mL, p = 0.003); increases in serum P4 concentrations were not statistically significant (p = 0.19). Three participants (20%) experienced increased luteal activity, with one presumptively ovulating post-rifampin (P4 = 15.8 ng/mL). ConclusionsWith only short exposure to a strong CYP3A inducer, ENG implant users experienced clinically significant decreases in serum ENG concentrations that led to changes in biomarkers indicative of waning suppression of ovulation. ImplicationsEven a short, 2-week course of treatment with rifampin places etonogestrel contraceptive implant users at risk for decreased contraceptive efficacy. Clinicians should counsel patients using etonogestrel implants considering any duration of rifampin therapy on the need for backup nonhormonal contraception or the use of an intrauterine device to avoid unintended pregnancies.

Expression profile of genes related to pregnancy maintenance in Dromedary Camel during the first trimester

So far, few signals involved in embryo-maternal dialogue have been identified in pregnant she-camel. Our objective was to investigate expression profiles of genes relevant to uterine extracellular matrix remodeling (ITGB4, SLCO2A1, FOS, and JUN), uterine tissue vascularization, and placental formation (VEGFA, PGF, and PDGFA), embryonic growth and development (IGF1 and PTEN), plus cell death of uterine tissue (BCL2) in early pregnant versus non-pregnant she-camels. Forty genital tracts (20 pregnant and 20 non-pregnant) and blood samples were collected from abattoirs. Total RNA was extracted from uterine tissues and qRT-PCR was conducted for candidate genes. Serum concentrations of progesterone (P4) and estradiol17-β (E2) were measured. Expression of ITGB4, FOS, and PGF genes increased (P < 0.001) in the right uterine horn of pregnant versus non-pregnant she-camels. Moreover, JUN, SLCO2A1, VEGFA, and PTEN mRNAs were up-regulated (P < 0.001) in various segments of uterine tissues in pregnant groups. The PDGFA transcript was over-expressed (P < 0.001) in both uterine horns of pregnant groups. Additionally, IGF1 was higher (P < 0.001) in the right horn and the uterine body of pregnant groups, and expression of BCL2 was increased (P < 0.001) in the pregnant uterine body. Moreover, serum concentrations of P4 were higher (P < 0.001) and E2 lower (P < 0.05) in pregnant she-camels. Taken together, the fine-tuning of genes related to implantation, matrix formation, vascularization, and placental formation is highly required for successful pregnancy in she-camels.

Cystic ovarian disease in dairy cattle: Diagnostic accuracy when using B-mode and color Doppler ultrasound

The most frequently reported definition of cystic ovarian disease in cattle is an abnormally persistent follicle (>7 to 10 d) with a diameter >25 mm. Discrimination between luteal and follicular ovarian cystic structures has traditionally been conducted by measuring the rim width of luteal tissue. The most common practice used in the field for diagnosis of cystic ovarian disease is examination by rectal palpation with or without the use of a B-mode ultrasound. Color Doppler ultrasound technology allows assessment of blood flow area measurements in the ovary, which has been proposed as a potential indirect measure for plasma progesterone (P4) concentrations. The objective of this study was to compare the diagnostic accuracy of differentiating luteal structures from follicular ovarian cysts using measures collected with B-mode and color Doppler transrectal ultrasonography. The definition of an ovarian cyst was a follicle greater than 20 mm in diameter in the absence of a corpus luteum that persisted for at least 10 d. A 3-mm luteal rim width was used to differentiate follicular and luteal cysts. A total of 36 cows were enrolled in the study during routine herd reproductive examination visits, with 26 and 10 having follicular and luteal cysts, respectively. Cows enrolled in the study were examined using a Mini-ExaPad mini ultrasound with color Doppler capabilities (IMV Imaging Ltd.). Blood samples were collected from each cow to measure P4 serum concentrations. History and signalment of each cow, including days in milk, lactation, times bred, days since last heat, milk composition, and somatic cell counts, were retrieved from an online database (DairyComp 305, Valley Agricultural Software). The accuracy of diagnosing follicular from luteal cysts based on luteal rim thickness was analyzed by receiver operating characteristic (ROC) curve using P4 as the gold standard, where P4 concentrations exceeding 1 ng/mL was defined as luteal, and all other structures with less P4 were considered follicular. Luteal rim and blood flow area were selected for further analysis because they presented the best ROC curves for differentiating cystic ovarian structures, with areas under the curve of 0.80 and 0.76, respectively. Luteal rim width of 3 mm was used as the cutoff standard in the study, resulting in sensitivity and specificity of 50% and 86%, respectively. Blood flow area of 0.19 cm2 was used as the cutoff standard in the study, resulting in sensitivity and specificity of 79% and 86%, respectively. When combining the use of luteal rim width and blood flow area to differentiate cystic ovarian structures, a parallel approach resulted in sensitivity and specificity of 73% and 93%, respectively, whereas an in-series approach resulted in sensitivity and specificity of 35% and 100%, respectively. In conclusion, the use of color Doppler ultrasonography when discriminating between luteal and follicular ovarian cysts in dairy cattle resulted in higher diagnostic accuracy compared with using B-mode ultrasonography alone.

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.

Effects of injectable and intravaginal progesterone on ewes' reproductive performance at breeding season beginning.

Progesterone (P4) is a steroid hormone that has a regulatory role in the female reproductive system. Studies on the effects of injectable progesterone on ewes are scarce, mainly related to their reproductive responses in the breeding season. This study aimed to compare reproductive performance and serum P4 concentration using injectable or intravaginal P4 in ewes. Two hundred and forty and eight Santa Inês x Dorper ewes (BW; 52.67 ± 11.76 kg; mean ± SE), body condition score (BCS; 2.5 ± 0.8; scale of 1-5), were distributed in four treatments: (i) Control: without administration of P4; (ii) CIDR: intravaginal implantation of 330 mg of P4 for 7 days; (iii) 1P4: 15 mg of P4 intramuscular (IM); and (iv) 2P4I: 30 mg of P4 IM. The first 18 days of breeding season were considered the synchronization period. Except for ewes in the control group, all other ewes received 263 μg IM of cloprostenol sodium for lysis of eventual CL at 24 h before the P4 treatment. After the synchronization period, all ewes were kept together with males for extra 28 days in the breeding season. At the beginning of breeding season, 90% of the ewes had serum P4 concentration less than 1 ng/mL. The estrus rate was greater (P < 0.01) in ewes on the CIDR treatment, with similar estrus rate among the other treatments. The P4 implant was able to keep blood P4 concentration greater than 1 ng/mL in ewes that received an implant of P4 during the 7 days. After implant removal, there was a great increase in the estrus manifestation on ewes in the CIDR treatment, leading to an increase in pregnancy rate at the beginning of breeding season. The current study demonstrated that ewes that received an intramuscular injection of 15 or 30 mg of P4 had similar reproductive performance than ewes that did not receive any P4 intramuscular injection. However, when ewes were implanted with P4 (CIDR), these animals had an increase in estrus manifestation, leading to greater pregnancy earlier during the breeding season.

Cysteine dioxygenase and taurine are essential for embryo implantation by involving in E2-ERα and P4-PR signaling in mouse

BackgroundTaurine performs multiple physiological functions, and the maintenance of taurine level for most mammals relies on active uptake from diet and endogenous taurine synthesis through its synthesis enzymes, including cysteine dioxygenase (CDO). In addition, uterus tissue and uterus fluid are rich in taurine, and taurine synthesis is regulated by estrogen (E2) and progesterone (P4), the key hormones priming embryo-uterine crosstalk during embryo implantation, but the functional interactions and mechanisms among which are largely unknown. The present study was thus proposed to identify the effects of CDO and taurine on embryo implantation and related mechanisms by using Cdo knockout (KO) and ovariectomy (OVX) mouse models.ResultsThe uterine CDO expression was assayed from the first day of plugging (d 1) to d 8 and the results showed that CDO expression level increased from d 1 to d 4, followed by a significant decline on d 5 and persisted to d 8, which was highly correlated with serum and uterine taurine levels, and serum P4 concentration. Next, Cdo KO mouse was established by CRISPER/Cas9. It was showed that Cdo deletion sharply decreased the taurine levels both in serum and uterus tissue, causing implantation defects and severe subfertility. However, the implantation defects in Cdo KO mice were partly rescued by the taurine supplementation. In addition, Cdo deletion led to a sharp decrease in the expressions of P4 receptor (PR) and its responsive genes Ihh, Hoxa10 and Hand2. Although the expression of uterine estrogen receptor (ERα) had no significant change, the levels of ERα induced genes (Muc1, Ltf) during the implantation window were upregulated after Cdo deletion. These accompanied by the suppression of stroma cell proliferation. Meanwhile, E2 inhibited CDO expression through ERα and P4 upregulated CDO expression through PR.ConclusionThe present study firstly demonstrates that taurine and CDO play prominent roles in uterine receptivity and embryo implantation by involving in E2-ERα and P4-PR signaling. These are crucial for our understanding the mechanism of embryo implantation, and infer that taurine is a potential agent for improving reproductive efficiency of livestock industry and reproductive medicine.

Analytical Validation of the IMMULITE® 2000 XPi Progesterone Assay for Quantitative Analysis in Ovine Serum.

Monitoring circulating progesterone (P4) concentration is an important component of basic and applied reproduction research and clinical settings. IMMULITE® 2000 XPi (Siemens) is a newly upgraded fully automated immunoassay system marketed for human use to measure concentrations of different analytes including P4. Our objective was therefore to characterize the analytical performance of the IMMULITE® 2000 XPi P4 immunoassay across the reportable range in ovine serum. This validation of analytical performance included determining (1) linearity, (2) precision through within-run, and between-run coefficient of variation (CV) calculations, (3) accuracy through bias calculations for spiking-recovery bias and interlaboratory (range and average based) bias for two laboratories across the reportable range (0.2-40 ng/mL). The average within-run and between-run precision (CV%) across the reportable range of the IMMULITE® 2000 XPi P4 immunoassay for serum P4 concentration were both &lt;5%, ranging between 2-8%. The average Observed Total Analytic Error (TEo) reported here for serum P4 concentration across the reportable range was ~30%, ranging from 14.8-59.4%, regardless of the considered bias. Based on these data we conclude that the automated IMMULITE® 2000 XPi P4 immunoassay provides a precise, accurate, reliable, and safe method for measuring P4 concentration ovine serum.

P079Etonogestrel contraceptive implant users can experience breakthrough ovulation with only two weeks of exposure to a strong cyp3a inducer

<h3>Objectives</h3> Rifampin is a strong CYP3A inducer and the gold standard for evaluating CYP3A-mediated drug-drug interactions per the US Food and Drug Administration. We hypothesized that even a short course (two weeks) of exposure would lead to breakthrough ovulation-mediated changes in endogenous estradiol (E2) and progesterone (P4) concentrations among etonogestrel (ENG) implant users. <h3>Methods</h3> We enrolled healthy people using ENG implants for 12–36 months. We measured baseline serum concentrations of E2 and P4 as surrogate biomarkers for ovulation suppression using clinical chemiluminescent immunoassays. After two weeks of rifampin 600 mg daily, we repeated E2 and P4 measurements. We determined breakthrough ovulation based on pre-exposure P4 3.0ng/mL with concurrent rise in post-treatment E2. <h3>Results</h3> Fifteen participants completed all study procedures. Participants' median age was 28.2 (range, 21.8–34.1), median BMI was 25.2 kg/m2 (range, 18.9–37.3), and median duration of implant use was 22 months (range, 12–32). At baseline, median E2 concentrations were 73 pg/mL (range, 32–686) and median P4 concentrations were 0.3 ng/mL (range, 0.2–6.1). After concomitant rifampin, median E2 concentrations were 201 pg/mL (range, 59–951) and median P4 concentrations were 0.4 ng/mL (range, 0.2–15.8). Serum E2 concentrations significantly increased with rifampin exposure (p=0.003, paired Wilcoxon signed rank test), but the increases in serum P4 concentrations were not statistically significant (p=0.185). Three participants (20%) experienced breakthrough ovulation with the following P4 changes: 0.3>5.0, 0.8>7.3, 0.5>15.8 (pre>post; ng/mL). <h3>Conclusions</h3> With only a short exposure to a strong CYP3A inducer, ENG implant users experienced clinically significant changes in hormone biomarkers indicative of waning follicular development and ovulation suppression.

Effect of increased serum progesterone on luteal morphology and function, pregnancy, and gestational loss in Nelore cows.

The aim of the present study was to determine the impact of different strategies for increasing the level of serum progesterone (P4) on luteal morphology and function in bovine females. The effects of increasing P4 on pregnancy rate and gestational loss (GL) in Nelore cows subjected to timed artificial insemination (TAI) were also evaluated. A total of 939 cows were divided into three groups: P4LA (n=305), 150 mg of long-acting injectable P4 7 days after TAI; GnRH (n=306), 10μg of buserelin acetate 7 days after TAI; and control (n=328), no hormone treatment after TAI. Doppler ultrasound assessments and P4 measurements were performed on days 7 and 16 after TAI. The pregnancy rate and GL as a function of treatment were compared using the SAS GLIMMIX procedure. Corpus luteum (CL) vascular perfusion, volume, and plasma P4 concentration were analysed using the SAS PROC MIXED procedure. No significant difference was found among the treatments in terms of volume, number of pixels, and CL intensity or in the serum P4 concentration at 7 days after ovulation. The CL blood flow at 16 days after ovulation was lower in the P4LA and GnRH groups than that in the control group (p < .01). Serum concentrations of P4 at 16 days after ovulation were higher in the P4LA and GnRH groups than those in the control group (p=.04). A difference in the pregnancy rate (p=.003) and a trend in GL (p=.07) as a function of treatment were found. Overall, long-acting injectable P4 supplementation on day 7 after TAI or GnRH administration affected CL vascularization and increased the serum concentrations of P4 16 days after ovulation, promoting better pregnancy rates than the control.

The Efficacy of a 3β-Hydroxysteroid Dehydrogenase Inhibitor for the Termination of Mid-Term Pregnancies in Dogs.

Progesterone (P4) is the only hormone needed to maintain pregnancy in dogs. Therefore, a competitive inhibitor of 3β-hydroxysteroid dehydrogenase (3β-HSD) could be a safe and effective option to terminate pregnancy by inhibiting P4 synthesis. To address this hypothesis, we investigated the efficacy of trilostane (TRL), a competitive inhibitor of 3β-HSD, in terminating pregnancy in dogs. Twenty-one dogs between days 30 and 38 of pregnancy were randomly assigned to one of two treatment groups (trilostane (TRL) and aglepristone (AGL)) and an untreated control (CON) group (n = 7 dogs each). Fetal heart rates (FHRs) (measured at 12 h intervals) and serum P4 concentrations (measured at 6 h intervals) were evaluated. The pregnancy termination rates were 0% and 100% in the TRL and AGL groups, respectively. The decrease in the FHR in the TRL and AGL groups was significantly lower than that observed in the CON group. There was a marked decrease in P4 concentrations in the TRL group 6, 54, and 102 h after the initiation of treatment. The luteal expression of StAR appeared to be weaker in the AGL group than the CON group. In conclusion, although a treatment-induced decrease was observed in plasma P4 concentrations, a seven-day TRL treatment alone was not effective in terminating pregnancies. Further studies are needed on the effects of the prolonged administration of TRL with varying doses and frequencies for the termination of mid-term pregnancy in dogs.

Influence Of The Menstrual Cycle On Muscle Damage Marker And Leukocyte Reaction Following Eccentric Exercise

PURPOSE:Ovarian hormone could inhibit exercise-induced muscle damage and leukocyte infiltration. Premenopausal women experience ovarian hormonal changes by the menstrual cycle (MC); therefore, MC could be related to muscle damage and leukocyte reaction, although it is not be clear. The purpose of this study was to investigate how MC could influence serum creatine kinase (CK) and leukocyte reaction following eccentric exercise. METHODS: Twenty-two untrained women with eumenorrhea (age 27.0 ± 6.4 years; height 160.1 ± 4.2 cm; weight 57.8 ± 7.0 kg; and body mass index 22.5 ± 2.3 kg/m2 [mean ± SD]) participated in the study. The subjects performed 10 sets of 6 repetitions of eccentric exercises with their nondominant arm in the early follicular phase (E-FP, n = 12) or midluteal phase (M-LP, n = 10). Blood samples were collected pre-exercise (PRE) and 4, 48, and 96 hours (h) post-exercise (POST) to analyze serum estradiol (E2) and progesterone (P4) concentration, serum CK, leukocyte count, and leukocyte fractionation (neutrophils, lymphocyte, monocyte, and other leukocytes). We assessed the isometric strength of elbow flexion, range of motion in the elbow joint, upper arm circumference and visual analog scale rating of perceived muscle soreness at PRE, immediately post, POST 4, 48, and 96 h. RESULTS: The serum E2 concentrations at PRE were significantly higher in the M-LP than the E-FP (E-FP 42.9 ± 18.6 vs. M-LP 186.4 ± 98.6 pg/mL; p < 0.01). The serum P4 concentrations at PRE were significantly higher in the M-LP than the E-FP (E-FP 0.2 ± 0.1 vs. M-LP 15.0 ± 3.1 ng/mL; p < 0.01). Significant time effects were present for serum CK at PRE to POST 48 h and 96 h (p < 0.05). No MC phase effect could be observed. The percent change in neutrophils at PRE to POST 4 h tended to be lower in the M-LP than the E-FP (E-FP 38.2 ± 32.6 vs. M-LP 11.4 ± 20.6%; p = 0.050). The serum E2 and P4 concentrations were not related to serum CK at all measurement points. A significant positive correlation could be observed between the PRE serum E2 concentration and PRE lymphocyte count (r = 0.470, p < 0.05). It was unrelated after the exercise. The POST 96 h serum CK was related to the POST 96 h neutrophil count (r = 0.469, p < 0.05). CONCLUSIONS: These results suggest that MC could affect the early leukocyte reaction following eccentric exercise without influencing the serum CK.

Form of dietary selenium affects mRNA encoding interferon-stimulated and progesterone-induced genes in the bovine endometrium and conceptus length at maternal recognition of pregnancy.

Widespread regions of the southeast United States have soils, and hence forages, deficient in selenium (Se), necessitating Se supplementation to grazing cattle for optimal immune function, growth, and fertility. We have reported that supplementation with an isomolar 1:1 mix (MIX) of inorganic (ISe) and organic (OSe) forms of Se increases early luteal phase (LP) concentrations of progesterone (P4) above that in cows on ISe or OSe alone. Increased early LP P4 advances embryonic development. Our objective was to determine the effects of the form of Se on the development of the bovine conceptus and the endometrium using targeted real-time PCR (qPCR) on day 17 of gestation, the time of maternal recognition of pregnancy (MRP). Angus-cross yearling heifers underwent 45-d Se-depletion then repletion periods, then at least 90 d of supplementation (TRT) with 35 ppm Se per day as either ISe (n = 10) or MIX (n = 10). Heifers were inseminated to a single sire after detected estrus (day 0). On day 17 of gestation, caruncular (CAR) and intercaruncular (ICAR) endometrial samples and the developing conceptus were recovered from pregnant heifers (ISe, n = 6 and MIX, n = 6). qPCR was performed to determine the relative abundance of targeted transcripts in CAR and ICAR samples, with the expression data subjected to one-way ANOVA to determine TRT effects. The effect of TRT on conceptus development was analyzed using a one-tailed Student's t-test. When compared with ISe-treated heifers, MIX heifers had decreased (P < 0.05) abundance of several P4-induced and interferon-stimulated mRNA transcripts, including IFIT3, ISG15, MX1, OAS2, RSAD2, DGAT2, FGF2 in CAR and DKK1 in ICAR samples and tended (P ≤ 0.10) to have decreased mRNA abundance of IRF1, IRF2, FOXL2, and PGR in CAR samples, and HOXA10 and PAQR7 in ICAR samples. In contrast, MIX-supplemented heifers had increased (P < 0.05) mRNA abundance of MSTN in ICAR samples and an increase in conceptus length (ISe: 17.45 ± 3.08 cm vs. MIX: 25.96 ± 3.95 cm; P = 0.05). Notably, myostatin increases glucose secretion into histotroph and contributes to advanced conceptus development. This advancement in conceptus development occurred in the presence of similar concentrations of serum P4 (P = 0.88) and whole blood Se (P = 0.07) at MRP.

Dietary organic trace mineral supplement affects steroid hormone, antioxidant enzyme concentrations, and reproductive performances in dairy cows

Supplementation of organic trace mineral (OTM) may affect reproductive functions in several aspects and has been studied, but very rare is reported in dairy cows. To determine the effects of dietary OTM supplement during pre- and postpartum with timed artificial insemination (TAI) protocol on steroid hormones, antioxidant enzymes, and reproductive performances in dairy cows. Prepartum Holstein cows (n = 60) were randomly assigned to received either a control diet or OTM supplemented for 42 days. Both groups were fed the same total mixed ration, but were supplemented with 5 g/head/d OTM (Bioplex®) in the treatment group. Blood and follicular fluid samples were collected to determine progesterone (P4), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and estradiol (E2) concentrations. Cows fed OTM had better placental expulsion period, milk yield, and reproductive performance parameters (day to first ovulation and estrus, percentage of first estrus and size of preovulatory follicle) than control cows (P &lt; 0.05). Cows fed OTM affected (P &lt; 0.05) the serum concentration of P4 on days 12, 15, 18, 21, and 42 after TAI. Follicular fluid concentrations of E2 in large follicle in OTM cows were greater than control cows (336.3 and 376.9 ng/mL; P &lt; 0.05). Cows supplemented with OTM had greater serum SOD and GSH-Px activities than control cows (15.5 vs. 10.6 U/mL and 12.1 vs. 9.9 U/mL; P &lt; 0.05). In conclusion, supplements with OTM improved uterine and placental health, steroid hormones concentrations, and antioxidant enzyme activities but did not improve conception rate in dairy cows.

Effects of human chorionic gonadotropin on the last day of a 5-day CIDR Synch protocol and 5 days later on circulating progesterone concentrations and luteal area in Holstein heifers

The objective was to determine the effect of replacing GnRH with human chorionic gonadotropin (hCG) on the last day of a 5-d CIDR Synch protocol (d 0) and inducing accessory corpus luteum (CL) formation with hCG 5 d later (d 5) on serum progesterone (P4) concentrations and luteal dynamics in dairy heifers. Holstein heifers (n = 207) were synchronized with a 5-d controlled internal drug release (CIDR) Synch protocol (d −8: used CIDR inserted; d −3: CIDR removed and PGF2α). Heifers were randomly assigned to 1 of 4 treatments on d 0: control (G; n = 55), H (n = 50), GH (n = 53), and HH (n = 49). Heifers in G were treated with 100 μg of GnRH on d 0, while H heifers received hCG (3,300 IU) on d 0. Heifers enrolled in GH were treated with GnRH on d 0 and hCG on d 5, while HH received hCG on d 0 and 5. Ovaries were scanned by ultrasound on d 0, 5, and 12, and blood was collected on d 0, 5, 7, and 12. Heifers that ovulated before or after the hCG or GnRH d 0 treatment and had P4 ≤ 0.50 ng/mL on d 0 were considered as synchronized. Overall protocol synchronization response was 93.8%, with no differences among treatments. Only synchronized heifers (n = 193) were included in the analyses of luteal dynamics after d 0. Serum P4 concentration and original CL luteal area on d 5 in heifers treated with hCG on d 0 (H + HH) were greater than in heifers treated with GnRH on d 0 (G + GH). Almost all heifers treated with hCG on d 5 had ≥2 CL on d 12 (98.6%). Ovulatory response for d 5 hCG treatment did not differ for GH versus HH (97.2 vs. 94.7%). Heifers in HH had the highest serum P4 on d 7, and G had the lowest serum P4 on d 7 and 12. In contrast, serum P4 on d 7 did not differ for H versus GH. On d 12, serum P4 and total luteal area were not different for GH versus HH. In summary, heifers that received hCG on d 0 had a larger total luteal area and greater serum P4 concentration on d 5 than heifers treated with GnRH on d 0. Moreover, hCG on d 5 promoted a greater proportion of heifers with ≥2 CL on d 12 and a larger luteal area of the original CL, which resulted in a larger total luteal area on d 12. The HH treatment successfully increased serum P4 concentrations in heifers on d 7 compared with the other treatments.

Effect of the addition of GnRH and a second prostaglandin F2α treatment on pregnancy per artificial insemination in lactating dairy cows submitted to an estradiol/progesterone-based timed-AI protocol

Two experiments determined whether the addition of GnRH at the beginning of an estradiol (E2)/progesterone (P4)- based synchronization protocol and/or a second dose of prostaglandin F2α (PGF2α) the day before P4 device removal improves pregnancy rate in lactating dairy cows. On Day 0, all cows received a CIDR-B device and 2 mg i.m. estradiol benzoate, and half received 200 μg i.m. gonadorelin acetate (GnRH). On Day 7, cows were further subdivided to receive PGF2α (500 μg i.m. cloprostenol) or no PGF2α treatment. On Day 8, CIDR-B were removed, and all cows received PGF2α, 1 mg estradiol cypionate and 400 IU eCG i.m., and an estrus detection aid. Experiment 1 was designed to evaluate the effect of treatments on follicular development from P4 device removal to ovulation, expression of estrus, time of ovulation and serum P4 concentrations. Cows (n = 76) were examined by ultrasonography and bled for serum P4 determinations every 12 h from the time of P4 device removal but were not inseminated. In Experiment 2, all cows (n = 1036) were inseminated based on estrus detection using tail-paint. Cows with >50% of the paint rubbed-off by 48 h after P4 device removal were inseminated at that time, whereas those not in estrus received 100 μg i.m. of GnRH and were inseminated 12 h later. In Experiment 1, the interval from P4 device removal to ovulation was 71.7 ± 1.5 h and did not differ among groups. However, cows that received 2 injections of PGF2α had a greater (P < 0.01) estrus rate and lower (P < 0.01) P4 concentrations at 48 h after P4 device removal than those that received 1 PGF2α (estrus rate: 86.8% vs 68.4% and P4 concentration: 0.12 ± 0.01 vs 0.36 ± 0.07, for 2 and 1 PGF2α, respectively). In Experiment 2, estrus rate was also influenced by the number of PGF2α treatments, regardless of whether cows received or did not receive GnRH on Day 0 (2 PGF2α: 84.7%, 438/517 vs 1 PGF2α: 65.7%, 341/519; P < 0.01). Furthermore, there was a GnRH treatment by number of PGF2α treatments interaction (P < 0.05) on P/AI that was attributed to greater (P < 0.05) P/AI in cows that received GnRH on Day 0 and 2 PGF2α than in the other three treatment groups (EB+1 PGF2α: 45.2%, 119/263; EB+2 PGF2α: 45.8%, 119/260; EB + GnRH + 1 PGF2α: 45.7%, 117/256 and EB + GnRH + 2 PGF2α: 57.2%, 147/257). It was concluded that the addition of GnRH on Day 0 and a second dose of PGF2α the day before P4 device removal improves P/AI in lactating dairy cows synchronized with an estradiol/P4-based protocol.

Impact of vitamin AD3E treatment before application of controlled intra-vaginal drug releaser on clinical and hematologic findings, ovarian hormones, and calving rates in she-camels (Camelus dromedarius).

The scope of using controlled intra-vaginal drug releasers (CIDR) in the camel industry was small relative to that of cow, sheep, and goat. The purpose of this study was to determine the effects of intramuscular injection of AD3E (25 ml/400 kg) as a premedication protocol before CIDR treatment on ovarian hormones profile and calving rate in dromedary she-camels (Camelus dromedarius) through measuring concentrations of progesterone (P4) and estradiol (E2) before, during, and after CIDR use. The study was conducted on she-camels (C. dromedarius) (n = 24) aged between 6 and 14 years. The animals were assigned into two equal groups: CIDR treated group (CIDRgr) and AD3E plus CIDR treated group (CIDR+ gr). Except for hemoglobin, the clinical and hematological findings showed no remarkable change either within each independent group or between CIDRgr and CIDR+ gr. Serum P4 and E2 concentrations differed significantly between the first 7 days (during CIDR use) and days from 9 to 15 (after CIDR use) in dromedary camels. Serum concentrations of P4 and E2 showed significant elevation at CIDR+ gr at day 0 compared with CIDRgr. This significant increase in serum of P4 and E2 at CIDR+ gr compared to those of CIDRgr was also observed either during CIDR use (days 3, 5, and 7) or after CIDR use (days 9, 11, 13, and 15). During CIDR use, serum concentrations of P4 were negatively correlated with those of E2 either in CIDRgr or CIDR+ gr, however, they were positively correlated after CIDR removal either in CIDRgror CIDR+ gr. Regarding the calving rate, it was highest in CIDR+ gr whereas CIDRgr showed the lowest one. It was concluded that the AD3E pretreatment protocol of she-camels before insertion of CIDR could be used successfully during the breeding season to improve fertility in she-camels whereas the serum ovarian hormones profile and calving rates were clearly changed due to pretreatment of she-camels with AD3E before CIDR insertion.

Progesterone Concentrations during Canine Pregnancy

Simple SummaryFor breeders and bitches alike, pregnancy is a challenging period. Insufficient serum progesterone concentrations have frequently been suggested to be responsible for pregnancy loss or early parturition, without any scientific evidence to support those claims. In this study, a large number of bitches was followed throughout pregnancy, and serum progesterone concentrations were determined to deduce which concentrations could be considered normal. Three blood samples were collected, one each during early, mid and late pregnancy. The results indicated that progesterone concentrations can be lower than previously described as adequate in the veterinary literature, with the bitch still exhibiting no signs of pregnancy distress and giving birth to healthy puppies at the expected time.Pregnancy and lactation are amongst the most challenging times of a bitch’s life. Most studies focusing on the endocrinological aspect of pregnancy consider only a small number of animals. The aim of this study was to evaluate progesterone (P4) concentrations in a large number of bitches during early, mid and late pregnancy. In total, 126 bitches of various breeds were recruited following a thorough clinical and gynecological examination during estrus. Blood samples were collected three times (T1–T3) during pregnancy or from non-pregnant dogs in diestrus, and P4 was measured via chemiluminescence. At T1 (11–19 days post-ovulation (dpo)), serum P4 concentrations were 30.23 ± 6.65 ng/mL and 28.45 ± 6.26 ng/mL, at T2 (23–32 dpo) they were 22.73 ± 6.27 ng/mL and 22.59 ± 5.77 ng/mL and at T3 (52–60 dpo) they were 6.68 ± 2.18 ng/mL and 3.17 ± 2.26 ng/mL, in pregnant (n = 98) and non-pregnant (n = 23) dogs respectively. The P4 concentrations differed significantly between pregnant and non-pregnant animals at the last examination (p ≤ 0.001). In the context of hypoluteoidism, the gathered data yielded interesting results. Overall, 28 out of 98 pregnant bitches showed a greater decline (>15 ng/mL) in P4 concentrations from early to mid-pregnancy, and 56 bitches showed P4 concentrations lower than deemed adequate (>20 ng/mL at T1 and T2, >5 ng/mL at T3) according to existing recommendations. Despite not being supplemented with P4, none of those animals suffered from abortion or preterm delivery. Considering that supplementation of P4 can entail considerable risks for the bitch and the puppies, more research on P4 concentration patterns, diagnosis of hypoluteoidism and treatment indications and options is indicated.

Blood perfusion and diameter of bovine corpus luteum as predictors of luteal function in early pregnancy.

A feasible and non-invasive luteal function assessment which enables timely intervention to improve progesterone (P4) support at early pregnancy is not well-established. The aim of this study was to determine the correlation among morphological and functional assessment methods of corpus luteum (CL) on Day 5 (D5) following timed-artificial insemination effect on luteal blood perfusion (LBP), CL diameter and serum P4 concentration. Beef heifers (n=89) were synchronized and inseminated (D0). On D5, CL was scanned by colour-Doppler ultrasonography using an LBP score (0 = absent; 3 = high blood perfusion); CL diameter was obtained and blood was collected. Diameter of CL had a positive linear effect on P4 concentration (p<.001); and larger CL diameter increased the probability to become pregnant (p<.05; odds ratio =1.21). Heifers with a CL larger than 14.95mm had a higher pregnancy rate than heifers with a 14.95mm or smaller CL (p<.05). Animals with LBP 2 and 3 had larger CL when compared to scores 0 and 1 (p<.001). Scores 1, 2 and 3 can accurately estimate heifers with higher P4 (accuracy =0.79, 0.72 and 0.61 respectively). In conclusion, LBP on Day 5 post-TAI allows to estimate heifers with higher P4 and larger CL size, and a larger CL diameter was positively associated with pregnancy rate.

The impact of the cavitary corpus luteum on the blood progesterone concentration and pregnancy rate of embryo recipient heifers

In cattle there are two types of the corpus luteum (CL): homogeneous (CLhom) and cavitary (CLcav). Although they are considered equal in their hormonal activity, the function of the CLcav is questioned by many veterinarians. In consequence, females with the CLcav are considered less valuable for assisted reproductive techniques such as embryo transfer (ET), where recipients with the CLhom are preferred. The aim of our study was to compare the two types of CLs regarding morphological endpoints, serum progesterone (P4) concentrations and final pregnancy rate (PR) in recipient heifers after ET.The morphological type of the CL and the final PR after ET of 432 Holstein Friesian heifers were analyzed. Oestrus was synchronized with two i.m. inj. of 0.5 μg cloprostenol 14 days apart. The ET took place on the day 7 of the estrous cycle, only animals with visible oestrum were chosen for the procedure. Clinical and transrectal US examinations of ovaries were performed on the day of ET. The presence of the CLhom or CLcav was determined, and the CL diameter and cross-sectional area were measured. If present, measurements of the cavities were also taken. Only embryos recovered immediately prior to the ET at the morula or blastocyst stage were transferred to the randomly chosen recipient that underwent initial selection regardless of the CL morphology. Additionally, from randomly selected heifers (N = 53, CLhom = 33; CLcav = 20) blood samples for serum P4 concentration analysis were collected. Pregnancy was determined by transrectal palpation 2 months after ET.The medium-sized CLcav was larger in diameter (P < 0.001) and cross-sectional area (P < 0.01) than the CLhom (mean ± SD) – 23.29 ± 3.6 mm and 419.57 ± 135.01 mm2 compared to 21.87 ± 3.57 mm and 384.73 ± 145.46 mm2, respectively. The mean diameter and cross-sectional area of the cavity were 10.2 ± 4.36 mm and 97.59 ± 71.13 mm2, respectively. The volume of both types of CLs was similar (P = 0.3). The mean serum P4 concentration was 8.84 ng/ml, higher (P < 0.0001) for females with the CLcav (11.31 ng/ml) than for those with the CLhom (7.15 ng/ml). The PR was 36.1%, higher (P < 0.05) for recipients with the CLcav (47.7%) compared to the CLhom (29.9%).The presence of a CLcav in the recipient heifers did not negatively affect the potential of the CL to maintain pregnancy. On the contrary, the CLcav may give the embryo better chances of surviving the time of pregnancy recognition and in consequence, may have a positive effect on PR in heifers.

Comparison of the effects of Presynch-Ovsynch, Presynch- Ovsynch+CIDR, and G6G protocols on the reproductive performance of lactating Holstein cows in the postpartum period

This study was performed on an industrial dairy farm near Tehran province, Iran, on 666 lactating Holstein cows from September 2019 to June 2020 in order to eliminate the harmful effects of heat stress on the reproductive performance of the cows. The hypothesis of the recent study was that by increasing the serum concentration of progesterone (P4) in the luteal phase before insemination in the form of Presynch-Ovsynch+CIDR (POC) and G6G protocols, the pregnancy rate of cows would be increased compared to the Presynch-Ovsynch (PO) protocol. The aim of the present study was to evaluate the effects of Presynch-Ovsynch (PGF2α-14d- PGF2α-12d-Ovsynch (OVS: GnRH(GnRH1)- 7d-PGF2α(PG)-2d-GnRH)-18h-Fixed-time artificial insemination (FTAI), n=212), Presynch-Ovsynch+CIDR (similar to the PO protocol, plus the use of a CIDR for 7 days, from the start of the OVS+FTAI protocol to the time of PG injection, n=230), and G6G (PGF2α- 2d-GnRH-6d-OVS+FTAI, n=224) protocols on the reproductive performance and pregnancy rate of lactating Holstein cows. The average body condition score (BCS) for the cows was about 2.5 at the time of the onset of the OVS+FTAI program. The average daily milk production for cows at the time of the start of the OVS+FTAI program was 38.3 kg/day. On average, the studied cows were inseminated 86 days postpartum. Implementation of the G6G protocol in comparison to the PO and POC protocols increased the serum concentration of P4 at GnRH1 (P=0.04). The cows that received the G6G protocol had a greater number of corpuses luteum (CL) on their ovaries at PG in comparison to the cows in the PO and POC protocols (P=0.03 and P=0.05, respectively). For all treatment protocols, the pregnancy rate of cows with &amp;gt;2 CLs on their ovaries at PG was significantly higher than for cows with ≤2 CLs on their ovaries at this time. The overall pregnancy rate for all cows studied was 42%. Separately, pregnancy rates for cows in PO, POC, and G6G protocols were calculated at 36.7%, 41.7%, and 47.3% respectively. The highest and the lowest pregnancy rates were obtained in the G6G and PO protocols, respectively (P=0.03). It can be concluded that the G6G protocol increased the pregnancy rate of cows in comparison to the PO but not to the POC protocol. This result can be attributed to the increase in serum concentration of P4 at GnRH1 and the greater number of CLs at PG following implementation of the G6G compared to the PO protocol. Although CIDR administration in the POC protocol led to an increase in the pregnancy rate of cows compared to the PO protocol, possibly by increasing the synchrony rate and the quality of the ovulatory follicle, this increase was not statistically significant (P=0.09).