Changes In Progesterone Concentrations Research Articles

Effects of repeated embryo flushing without PGF2α administration on luteal function, percentage of unwanted pregnancy and subsequent fertility in mares.

PGF2α is commonly given at the end of embryo flushing (EF) to shorten the interval to the next oestrus and ovulation. To determine the effect of repeated EF on plasma progesterone concentration, percentage of mares with endometritis, unwanted pregnancy and subsequent fertility in mares flushed without the use of PGF2α. Controlled experiments. Nine mares were inseminated in seven consecutive cycles (n = 63), to either perform an EF (n = 54) 7-9 days after ovulation or left pregnant (n = 9). PGF2α was not used to induce oestrus. Ultrasound examination and blood sampling were performed just before the EF and 72 h later to determine changes in progesterone concentration and signs of endometritis. The overall percentage of positive EF/pregnancy was 55.5% (30/54) and 66.7% (6/9), respectively. The likelihood of pregnancy/positive EF in the first three cycles was 55.5% (15/29). This was not different (p > 0.1) from the fertility of the last four cycles (69.4%, 25/36). In five EF cycles (9.3%), mares had signs of endometritis and early luteolysis (progesterone <2 ng/mL) 72 h after EF. The reduction in progesterone concentration by 72 h after EF was greater (p < 0.05) for Day 9 (-2.3 ± 0.7 ng/mL) than Day 7 (-1.0 ± 0.8 ng/mL) or Day 8 (-1.3 ± 1.1 ng/mL) cycles. The progesterone concentration in non-flushed mares did not vary significantly during the sampled period (Day 7-12). There were 5 cycles in which the donor mare remained pregnant after the EF, although four were from a single mare. The mare population was limited to barren and maiden mares. The cycle order and operator allocation to each EF were not randomised. EF induces a subtle, but significant reduction in progesterone concentrations compared with non-EF cycles. However, the percentage of mares with EF-induced full luteolysis is low (9.3%). The fertility of mares after repeated EF without administration of PGF2α was unaffected; however, there is a considerable risk of unwanted pregnancy (5/27 = 18.5%) in donors from which an embryo was not recovered.

MONITORING OF REPRODUCTION AND INFLUENCE OF FECAL PROGESTERONE ON REPRODUCTIVE CYCLICITY IN CAPTIVE SRI LANKAN SAMBAR DEER (RUSA UNICOLOR UNICOLOR)

This study examines the length of the estrous cycle in 16 Sambar deer hinds in National zoological gardens in Dehiwala and Kegalle, Sri Lanka (NZGDK) assessed with the use of changes in progesterone concentrations, along with the changes in the profile of this hormone and by the visual estrus manifestations. The objectives of the present study were to characterize ovarian activity throughout the estrous cycle and the non-pregnant luteal phase of captive sambar deer in Sri Lanka. These objectives were achieved with the use of radioimmunoassay (RIA) to measure fecal concentrations of progesterone and visual estrus manifestation. Fecal samples were collected from non-pregnant sambar deer hinds (aged 2–4 years)over the period of six months on daily basis, both during breeding and non-breeding seasons. Estrous cycles were recorded in non-pregnant females, based on fecal progesterone concentrations. The average estrous cycle length was 26.1±2.08 days (mean ± SEM) and 2.10 ± 0.51 days in the inter-luteal phase.The average fecal progesterone concentrations attained the peak mid-luteal values of 2.74 ng mL–1. There appeared to be variation in fecal progesterone amplitude between animals and between dates, but the low frequency of sampling prohibited confirmation of trends. Behavioral estrus was detected only when the average progesterone concentrations were less than 0.07 ng mL–1. However, not all periods of depressed progesterone secretion were associated with the observed estrus. Behavioral estrus was detected in hinds when progesterone concentrations were less than 0.07 ng mL–1; a subsequent rise in progesterone indicated ovulation taking place at this time.

Dynamic Changes in Progesterone Concentration in Cows' Milk Determined by the At-Line Milk Analysis System Herd NavigatorTM.

Simple SummaryAccording to the literature, the at-line progesterone monitoring system Herd NavigatorTM (Lattec I/S, Hillerød, Denmark) was used in combination with a DeLaval milking robot (DeLaval Inc., Tumba, Sweden). It works automatically and provides real-time physiological information about lactating dairy cows. For making farm-management decisions, it is not only a novel tool for scientific research, but also a mechanism for improving productivity, food safety, animal well-being, the environment, and the public perception of the dairy industry. It has been hypothesized that the progesterone concentration determined by the at-line milk analysis system and changes in its dynamics correlate with the parity, reproductive status, and milk yield of cows. The aim of the current study was to evaluate relative at-line milk progesterone (mP4) dynamic changes, according to the parity and status of reproduction, and to estimate the relationship with productivity in dairy cows. Frequent automated mP4 sampling can help identify characteristics of mP4 dynamic changes associated with successful pregnancies, pregnancy losses, and potential differences in mP4 dynamics among parity groups, which have not been studied previously.The aim of the current instant study was to evaluate relative at-line milk progesterone dynamic changes according to parity and status of reproduction and to estimate the relationship with productivity in dairy cows by at-line milk analysis system Herd NavigatorTM. According to the progesterone assay, experimental animals were divided into three periods: postpartum, after insemination, and pregnancy. In the first stage of the postpartum period, progesterone levels in milk were monitored every 5 days. This period of reproductive cycle recovery was followed for 30 days (days 0–29). The second stage of the postpartum period (30–65 days) lasted until cows were inseminated. In the period (0–45 days) after cow insemination, progesterone levels were distributed according to whether or not cows became pregnant. For milk progesterone detection, the fully automated real-time progesterone analyzer Herd NavigatorTM (Lattec I/S, Hillerød, Denmark) was used in combination with a DeLaval milking robot (DeLaval Inc., Tumba, Sweden). We found that an at-line progesterone concentration is related to different parities, reproductive statuses, and milk yield of cows: the 12.88% higher concentration of progesterone in milk was evaluated in primiparous cows. The average milk yield in non-pregnant primiparous cows was 4.64% higher, and in non-pregnant multiparous cows 6.87% higher than in pregnant cows. Pregnancy success in cows can be predicted 11–15 days after insemination, when a significant increase in progesterone is observed in the group of pregnant cows.

Normal progesterone profiles during estrus in the bitch: A prospective analysis of 1420 estrous cycles

Appropriate timing of insemination in the bitch, through monitoring of serial serum progesterone concentrations, has historically been based on the detection of important physiological reproductive events which include the LH surge and ovulation 2–3 days later. Previous hormone profile studies during the estrus period in the bitch, in which determination of these physiological reproductive events has been defined, have been based on low numbers of bitches of similar breeds and body weights. Therefore, the aim of this large-scale prospective study was to define the normal serum progesterone profile during the estrus period in a large number of bitches of various breeds and body weights. In addition, we investigated if the rate of change in progesterone concentrations during the fertile period affected the reproductive performance of bitches after insemination with either fresh or frozen-thawed semen. A total of 1300 individual bitches, representing 84 different breeds, contributed 1420 estrous cycles and 4213 serum progesterone values over the 11-year (2007–2017) study period. The mean (±SD) progesterone concentration at estimated LH0 was 2.7 ± 0.6 ng/ml and at the time of estimated ovulation it was 4.8 ± 0.9 ng/ml and 7.2 ± 1.3 ng/ml (LH+2 and LH+3 respectively). There was no difference in the shape of progesterone profiles for bitches of different body weights. Furthermore, mean progesterone concentrations on each day between LH-6 and LH+7 were not different amongst bitches of different breeds and body weights and there was no effect of mean progesterone concentrations on any day on whelping rate or litter size. However, there was a significant effect of the rate of change in progesterone concentrations on litter size when frozen semen was used. The litter sizes of bitches inseminated with frozen semen with slow progesterone curves were significantly smaller compared to bitches with fast progesterone curves (3.9 ± 1.8 vs 5.6 ± 3.1 pups per litter respectively; P < 0.001). There was no effect of bitch age on the normal progesterone curve, or its rate of change. This is the first report of the normal progesterone profile during estrus in the bitch derived from a large number of serial progesterone measurements obtained from bitches of various breeds and body weights. Importantly, the predictability and reliability of this progesterone profile regardless of breed or body weight gives clinicians the confidence to accurately determine the optimal time for insemination, which is critical to reproductive success, especially when frozen semen is used.

Insights into the role of estrogen-related receptors α, β and γ in tumor Leydig cells

In this study, we demonstrate, for the first time, estrogen-related receptor (ERR) regulation of the physiological and biochemical status of testicular tumor Leydig cells. In a mouse tumor Leydig cells, ERRs (α, β, and γ) were silenced via siRNA. Cell morphology and cell physiology (proliferation and observation of monolayer formation) were performed by inverted phase-contrast microscope. Leydig cell functional markers (steroid receptors and signaling molecules) were examined by immunofluorescence and Western blotting. Additionally, progesterone secretion was assessed. Mitochondrial mass and membrane potential were analyzed by flow-cytometry while cGMP and Ca2+ concentrations were analyzed using immunoenzymatic and colorimetric assays, respectively. These results revealed, ERRs indirectly regulate Leydig cell proliferation while ERRα and β affect cell monolayer formation. ERRs interact with canonical and membrane estrogen receptors (ERα, ERβ, and GPER), androgen receptor, metalloproteinase (MMP 9), protein kinase A (PKA), extracellular-regulated kinase (ERK), and neurogenic locus notch homolog protein 2 (Notch2). Depending on the type of ERR knocked down, coupled with estradiol treatment, changes in progesterone concentration and cGMP and Ca2+ concentrations constitute a microenvironment that may effect tumor Leydig cell characteristics. ERRs should be considered important factors in developing of innovating approaches that target pathological processes of testicular Leydig cells.

A specific profile of luteal phase progesterone is associated with the development of premenstrual symptoms

There is a consensus that the development of premenstrual dysphoric states is related to cyclical change in gonadal hormone secretion during the menstrual cycle. However, results from studies seeking to link symptom severity to luteal phase progesterone concentration have been equivocal. In the present study we evaluated not only the absolute concentrations of progesterone but also the kinetics of the change in progesterone concentration in relation to development of premenstrual symptoms during the last 10days of the luteal phase in a population of 46 healthy young adult Brazilian women aged 18–39 years, mean 26.5±6.7years. In participants who developed symptoms of premenstrual distress, daily saliva progesterone concentration remained stable during most of the mid-late luteal phase, before declining sharply during the last 3days prior to onset of menstruation. In contrast, progesterone concentration in asymptomatic women underwent a gradual decline over the last 8days prior to menstruation. Neither maximum nor minimum concentrations of progesterone in the two groups were related to the appearance or severity of premenstrual symptoms. We propose that individual differences in the kinetics of progesterone secretion and/or metabolism may confer differential susceptibility to the development of premenstrual syndrome.

Organotin compounds cause structure-dependent induction of progesterone in human choriocarcinoma Jar cells

Organotin compounds, such as tributyltin (TBT) and triphenyltin (TPT), are typical environmental contaminants and suspected endocrine-disrupting chemicals because they cause masculinization in female mollusks. In addition, previous studies have suggested that the endocrine disruption by organotin compounds leads to activation of peroxisome proliferator-activated receptor (PPAR)γ and retinoid X receptor (RXR). However, whether organotin compounds cause crucial toxicities in human development and reproduction is unclear. We here investigated the structure-dependent effect of 12 tin compounds on mRNA transcription of 3β-hydroxysteroid dehydrogenase type I (3β-HSD I) and progesterone production in human choriocarcinoma Jar cells. TBT, TPT, dibutyltin, monophenyltin, tripropyltin, and tricyclohexyltin enhanced progesterone production in a dose-dependent fashion. Although tetraalkyltin compounds such as tetrabutyltin increased progesterone production, the concentrations necessary for activation were 30–100 times greater than those for trialkyltins. All tested active organotins increased 3β-HSD I mRNA transcription. We further investigated the correlation between the agonistic activity of organotin compounds on PPARγ and their ability to promote progesterone production. Except for DBTCl2, the active organotins significantly induced the transactivation function of PPARγ. In addition, PPARγ knockdown significantly suppressed the induction of mRNA transcription of 3β-HSD I by all active organotins except DBTCl2. These results suggest that some organotin compounds promote progesterone biosynthesis in vitro by inducing 3β-HSD I mRNA transcription via the PPARγ signaling pathway. The placenta represents a potential target organ for these compounds, whose endocrine-disrupting effects might cause local changes in progesterone concentration in pregnant women.

The Buserelin Enigma; How Does Treatment with this GnRH Analogue Decrease Embryo Mortality?

The use of the GnRH analogue buserelin administered in mid dioestrus has been shown to increase early pregnancy rate, reduce embryo loss and increase litter size in several species. The mechanism by which this works is unclear and may differ between species. In cattle it is believed that buserelin, by reducing oestrogen concentrations, causes the development of endometrial oxytocin receptors to be delayed. Luteal oxytocin binds with these receptors to promote the formation and secretion of prostaglandin. Any delay in the release of PGF2α would allow an underdeveloped embryo extra time to produce and release the maternal recognition of pregnancy signal, which might otherwise be lost following luteolysis. This may also be the mechanism in sheep and pigs although an increase in progesterone concentrations could also be important. However in the mare, following buserelin administration at 9 or 10 days after the detection of ovulation, pregnancy rates are already increased by 12-13 days compared with untreated controls, i.e. before the time of luteolysis, without any change in progesterone concentrations. In addition to an increased pregnancy rate, the twin embryo rate is also increased and subsequent embryo loss rate reduced. The possible mechanisms by which this may result in the mare are discussed.

Effect of Body Condition Score and Parity on Resumption of Postpartum Ovarian Activity in Montberliard Dairy Cows in Algerian Semi -Arid Area

A study was undertaken to evaluate the relationships between body condition score (BCS), parity and serum hormonal profiles (progesterone (P4) and estradiol -17β (E2)), and resumption of postpartum ovarian cyclicity on (primiparous (n=19) and multiparous (n=59) Montbeliard cows. The BCS was measured at calving, using a 1 to 5 point scale according to the Edmonson (1989) method. Blood samples were collected by jugular venous at 30 d, 40 d, 50 d and 60 d postpartum (10 days of interval). Serum hormonal analysis were determined by chimiluminescent Immuno-Assay (CLIA). Changes in progesterone concentration were exploited to assess the resumption of ovarian cyclicity. The results showed a significant increase in progesterone and estradiol -17β serum levels during postpartum period. The correlation analysis showed that P4concentration in cows was positively correlated with their BCS at 30 d, 40 d, 50 d and 60 d postpartum (r =0.630; r = 0.603; r = 0.622 and r = 0.467 respectively; P<0.01). Additionally, the BCS at calving was positively correlated with estradiol -17β concentration at 30 d (r= 0.628); 40 d (r= 0.612); 50 d (r= 0.444) and 60 d (r=0.294) (P<0.01). Furthermore, BCS at calving was strongly but negatively correlated with CCI (r= 0.704; P<0.01). The results of the current study provide an evidence of a negative correlation between calving conception interval and secretion of P4 and E2.

Detection of early pregnancy-specific proteins in Holstein milk

Bovine pregnancy is commonly diagnosed by rectal palpation or ultrasonography and changes in progesterone concentration. To determine a simpler and less expensive diagnostic method, we sought to identify early pregnancy-specific proteins in bovine milk by comparing samples collected from pregnant and non-pregnant Holstein cattle. Of the 600-700 protein spots visible on 2-DE gel images, 39 were differentially expressed in milk from pregnant and non-pregnant cattle. Antibodies generated against synthetic peptides of milk whey proteins expressed specifically during pregnancy were used to confirm protein expression patterns. Western blot analysis showed that the levels of expression of lactoferrin (lactotransferrin) and alpha1G T-type calcium channel subunit (alpha-1G) were higher in samples from pregnant than non-pregnant cattle. These findings suggest that assays for pregnancy-specific milk proteins may be used to diagnose pregnancy in cattle.

Alterations in systemic concentrations of progesterone during the early luteal phase affect RBP4 expression in the bovine uterus

Systemic progesterone affects the timing and duration of uterine endometrial gene and protein expression and has significant effects on conceptus development. The objective of the present study was to examine how changes in progesterone concentrations during the early luteal phase affect retinol-binding protein (RBP4) mRNA and protein concentrations in the uterus. Endometrial tissue and uterine flushings were recovered on Days 7 and 13 of the oestrous cycle in heifers with high, normal and low progesterone concentrations. RBP4 mRNA and protein concentrations were higher (P<0.05) on Day 13 compared with Day 7 in heifers with high and control progesterone concentrations. However, there was no difference in RBP4 protein concentrations between Days 7 and 13 in heifers with low progesterone (P>0.05). On Day 7, although heifers with low progesterone had lower RBP4 mRNA expression compared with controls (P<0.05) there was no difference in protein concentrations between treatment groups. On Day 13, RBP4 mRNA was 2-fold higher (P<0.001) in heifers with high and control progesterone compared with their low-progesterone counterparts and RBP4 protein concentrations were over 2-fold higher (P<0.001) in heifers with high compared to low progesterone. In conclusion, progesterone modulates uterine RBP4 mRNA and protein abundance in a time- and concentration-dependent manner.

Role of Luteinizing Hormone in Changes in Concentrations of Progesterone and Luteal Blood Flow During the Hours of a Simulated Pulse of 13,14-Dihydro-15-Keto-Prostaglandin F2alpha (PGFM) in Heifers

A bolus treatment (e.g., 25 mg) of prostaglandin F(2alpha) (PGF) in the study of luteolysis in cattle results in dubious interpretations. Therefore, in experiment 1 of the present study, a 13,14-dihydro-15-keto-PGF (PGFM) pulse was simulated by incremental intrauterine (IU) infusion of PGF for 2.7 h on Day 14 postovulation. Concentrations of PGFM during the first hour of infusion and at the maximum were not different between simulated (n = 7) and spontaneous (n = 7) pulses. In experiment 2, four groups (n = 6 per group) were treated at Minute 0 (beginning of infusion) as follows: saline (infused IU), PGF (infused IU), acyline/saline, and acyline/PGF. Two hours before Minute 0, each heifer was given flunixin meglumine to inhibit endogenous PGF secretion, and heifers in the acyline/saline and acyline/PGF groups were given acyline to inhibit luteinizing hormone (LH). Plasma progesterone concentrations were similar among groups during Minutes 0 to 60, with no indication of an initial transient progesterone increase in the two PGF groups. Progesterone began to decrease in the PGF groups at Minute 60 and to rebound at Minute 135 after the PGFM peak at Minute 120. The rebound was complete in association with an increase in LH in the PGF group, but it was not complete when LH was inhibited in the acyline/PGF group. Luteal blood flow increased during PGF infusion in the two PGF groups and remained elevated for approximately 2 h after the PGFM peak in the PGF group but not in the acyline/PGF group. Novel findings were that an initial transient increase in progesterone did not occur with the simulated PGFM pulse and that LH stimulated a progesterone rebound and maintained the elevated luteal blood flow after the PGFM peak.

36 CHANGES IN PLASMA CONCENTRATIONS OF PROGESTERONE AND ESTROGENS DURING GESTATION IN COWS WITH STILLBORN SOMATIC CELL CLONED CALVES

Cloning of mammals by nuclear transfer frequently results in gestational or neonatal failure with a variety of abnormalities that are likely caused by inappropriate epigenetic reprogramming. Early diagnosis of fetal abnormality is important for efficient production of cloned animals. Sex steroids are produced in the bovine placenta and their levels in the blood might be useful as a measure of fetal well being, as they are in humans. The objective of this study was to investigate whether changes in peripheral concentrations of progesterone and estrogens reflect fetal conditions. Donor cells for nuclear transfer were obtained from subculture of cumulus cells retrieved from ovarian follicles of a Japanese Black cow. Recipient oocytes were derived from ovaries obtained at an abattoir and matured in vitro. Metaphase II oocytes were enucleated and each fused with a donor cell by DC pulses. Nuclear transferred oocytes were activated and cultured for 7 days. Embryos developed to the blastocyst stage were transferred into the uterine horn ipsilateral to the ovary bearing the CL of 29 multiparous Japanese Black and Holstein crossbred cows at 7 to 8 days after the day of standing oestrus (Day 0). Blood was collected at regular intervals from Day 40 until parturition. Plasma progesterone, estrone, oestradiol-17β, and estrone sulfate were measured by RIA in 4 recipient cows. 2 vaginally delivered healthy calves weighed 35 and 36 kg on days 278 and 280, respectively. The other 2 delivered stillborn calves weighed 42 and 31 kg by Caesarean section on days 267 and 287, respectively. Steroid profiles were compared with each other and with those in a cow made pregnant by embryo transfer. Statistical differences at stages of gestation were analysed with repeated-measures ANOVA. Stillborn calves were subjected to necropsy and histopathological analysis. Significant differences in steroid levels were observed individually and temporarily. In the cow bearing the former stillborn, progesterone and oestradiol-17β concentrations tended to be lower during mid and late gestation, respectively. Large offspring syndrome, ventricular septal defect, pulmonary oedema, hepatic fibrosis, and placental dysplasia were found in the calf by necropsy and histopathological analysis. In the cow bearing the second stillborn, plasma progesterone concentrations were temporarily increased from days 220 to 230 and reached a peak of 16 ng mL–1, which was approximately 3 times as high as before (5.0 ± 0.9; mean ± s.d.), then followed by a similar transient increase in estrogens during days 240 and 250 (170 pg mL–1, 16 pg mL–1, and 21 ng mL–1 at peak of estrone, oestradiol-17β, and estrone sulfate, respectively). Thereafter, oestrogen concentrations stayed low until parturition. As internal organs of the stillborn calf were severely autolysed with polyhydramnios at birth, the fetus might have been dead around Day 250. These results imply that changes in peripheral steroid concentrations in some cases reflect fetal condition.

Intrapulse changes in progesterone and LH concentrations and luteal blood flow during an estradiol-induced pulse of a metabolite of prostaglandin F2α in heifers

Physiologic doses (0.1 mg followed by 0.05 mg 1 h later) of estradiol-17β in sesame-oil vehicle or vehicle alone ( n = 6/group) were given to heifers on day 14 after ovulation (preluteolysis) to study the effect of estradiol (E2) on circulating 13,14-dihydro-15-keto-PGF2α (PGFM) and on PGFM pulses. Blood samples were collected hourly for 10 h after the 0.1-mg treatment. The E2 group had an increase ( P < 0.03) in PGFM concentration by 4 h and greater ( P < 0.0001) prominence of the PGFM pulses. Changes in progesterone and LH concentrations and luteal blood flow within a PGFM pulse were characterized. Within a PGFM pulse in the E2 group, progesterone decreased ( P < 0.04) between Hours −2 and 0 (ascending portion; Hour 0 = peak of pulse) and increased ( P < 0.04) between Hours 0 and 2 (descending portion). Intrapulse changes in progesterone were not detected in the vehicle group. Concentration of LH in the E2 group increased ( P < 0.05) between Hours −1 and 1 and in the vehicle group decreased ( P < 0.03) between Hours −1 and 0 and increased ( P < 0.06) between Hours 0 and 1. The percentage of luteal area with blood-flow signals increased ( P < 0.02) between Hours 0 and 1 in the E2 group, and there were no other differences between hours in either group. Results were consistent with reports of changes in progesterone concentration within a PGFM pulse in cattle and demonstrated intrapulse changes in LH concentration and luteal blood flow during an E2-induced PGFM pulse.

Stimulation of pulses of 13,14-dihydro-15-keto-PGF2α (PGFM) with estradiol-17β and changes in circulating progesterone concentrations within a PGFM pulse in heifers

A single physiologic dose (0.1 mg) of estradiol-17β in sesame-oil vehicle or vehicle alone (n = 8) was given to heifers on day 14 after ovulation to study the effect on circulating 13-14-dihydro-15-keto-PGF2α (PGFM), PGFM pulses, and changes in progesterone concentrations within a PGFM pulse. Blood samples were collected hourly for 16 h after treatment. The estradiol group had a greater mean concentration of PGFM, greater number of heifers with PGFM pulses and number of pulses/heifer, and greater prominence of the PGFM pulses. Changes in progesterone concentrations were not detected during the 16 h sampling session in the vehicle group, indicating that the heifers were in preluteolysis. Progesterone decreased after 12 h in the estradiol group, indicating a luteolytic effect of the estradiol-induced PGF secretion as represented by PGFM concentrations. Intrapulse changes in progesterone were detected during a PGFM pulse in the estradiol group (P < 0.006), but not in the vehicle group. Progesterone increased (P < 0.01) between Hours −2 and −1 of an estradiol-induced PGFM pulse (Hour 0 = peak of pulse), decreased (P < 0.004) between Hours −1 and 0, and increased (P < 0.01) or rebounded between Hours 0 and 1. Results were compatible with previous reports of a role for estradiol in the induction of PGFM pulses in cattle and demonstrated intrapulse changes in progesterone concentrations during an induced PGFM pulse.

Controlled Delivery of a GnRH Agonist by a Silastic Implant (Gonazon) Results in Long‐Term Contraception in Queens

Many female cats are spayed to prevent problems associated with calling and unwanted pregnancies. This study describes the safety and efficacy of an alternative approach, using an azagly-nafarelin containing implant (Gonazon) inserted subcutaneously in the neck of six treated queens for 3 years. These six queens together with six controls were permanently housed with vasectomized tom cats, and changes in progesterone concentrations were used to document the contraceptive efficacy of Gonazon. All six control queens ovulated regularly throughout the treatment period (3 years), as shown by regular changes in progesterone concentration. Sixteen ovulatory cycles were observed in each control throughout the study. In Gonazon treated queens, during the week following implant placement, two queens displayed a treatment-induced rise in progesterone concentration. Later on, all treated queens continuously displayed low progesterone concentrations until 3 years post-implant insertion, with the exception of a single isolated episode (at approximately 2.5 years of treatment), of follicular luteinization in two queens. In all queens, azagly-nafarelin concentrations peaked in the week following implant insertion remained high for 1 month and later decreased slowly. After 2.5 years of treatment, azagly-nafarelin concentrations were still greater than 150 pg/ml in 3/6 queens. During a 6-month long extension of the study (36-42 months post-treatment), all queens (treated and controls) were run with intact tom cats. None of them conceived. Following autopsy, ovarian weight and diameter of the uterine horns of 3/6 treated queens were shown to be similar to those of the controls. In conclusion, this study demonstrated that Gonazon efficiently prevented ovulation in queens (100%) for 3 years. Return to fertile heat was not observed towards the end of treatment. However, in half of the treated queens, reversibility of the treatment induced effects on the genital tract was demonstrated.

A compartmental model describing changes in progesterone concentrations during the oestrous cycle

The objective of this study was to develop a mathematical model that accurately describes the rise and decline in plasma progesterone concentrations, and is able to define parameters describing progesterone appearance and disappearance during the bovine oestrous cycle. Daily plasma progesterone data from 27 cows were used to develop a compartmental model consisting of an appearance function and an appearance modulating function. Model outputs included an apparent appearance or secretion duration, appearance rate and an average disappearance rate (expressed as arbitrary units per day; units/d). Shape-based clustering identified three common shape-based groups (or clusters) of progesterone profiles defined as either 'peaked' profile, with the profile reaching a distinguishable peak, 'structured', with the profile exhibiting a wave-like pattern, or 'flat top', with the profile reaching a plateau. Differences in the model parameters for the three different shapes of progesterone profiles were examined: peaked (n=13), flat top (n=7) and structured (n=7). The mean duration of apparent appearance was 11.49 (SD 0.17 d) for all 27 profiles. The model estimates for total appearance of progesterone (area under the curve; ng/ml per cycle), mean appearance rate and maximum appearance rate were 69.04 ng/ml per cycle (SD 15.2 ng/ml per cycle), 3.19 ng/ml per cycle (sd 0.7 ng/ml per d) and 6.70 ng/ml (SD 1.31 ng/ml), respectively. The average disappearance rate was 1.0 units/d (SD 0.04 units/d). The apparent appearance duration was greatest (P<0.01) in the flat top profiles (12.54, SD 0.41 d) followed by the structured (11.77, SD 0.66 d) and the peaked (10.80, SD 0.30 d) profiles. Total and mean progesterone appearance, maximum progesterone appearance rate, and the progesterone disappearance rates were not different between the profiles. The model successfully simulated all components of the progesterone profile and was able to define specific parameters of different shaped progesterone profiles. A simple model able to estimate parameters describing progesterone appearance and disappearance can be used to explore the relationships between profile shapes and reproductive outcomes.

Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers

Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers ( n = 197, ∼2-years old) and they were inseminated 12–18 h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n = 69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n = 64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n = 64. Body condition (BCS) and locomotion scores (scale of 1–5) were recorded for all animals. Animals with a locomotion score ≥4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased ( P < 0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship ( P < 0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 ( P < 0.06). There was also a positive relationship ( P < 0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher ( P < 0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive association between changes in progesterone concentration during the early luteal phase and embryo survival rate. Supplementation with progesterone had no effect on either CL weight or conceptus size in pregnant animals. Lameness had a significant negative effect on early embryo survival.

Content of Steroid Hormones in the Blood and Adrenal Glands of Mice in the Dynamics of BCG- and SiO2-Induced Granulomatous Inflammation

Individual or combined administration of BCG vaccine and silicon dioxide to male mice induced a nonspecific stress response of the adrenal glands and gonads judging from changes in the concentration of blood cortisol and testosterone. The dynamics of cortisol concentration in the adrenal glands was similar, while changes in progesterone concentration were in antiphase to those in the blood. After combined administration of both inductors of granulomatous inflammation, changes in the concentrations of the studied hormones to a greater extent corresponded to their dynamics after injection of BCG.

Noninvasive Measurement of Fecal Progesterone Concentration in Toy Poodles by Time Resolved Fluoroimmunoassay (TR-FIA)

Progesterone is an important reproductive hormone and measurement of its level by repeated blood samplings is beneficial to monitoring of estrus cycle. However, since toy poodles have a small body size and thin-walled blood vessels, repeated blood samplings cause stress and affect their preparation for mating or artificial insemination (AI). Therefore, a noninvasive method for monitoring progesterone concentration should be developed. Here, we show that time-resolved fluoroimmunoassay (TF-RIA) is a useful noninvasive method for determining the progesterone concentration in serum and fecal samples obtained from toy poodles. Present results demonstrate that progesterone concentrations in the fecal correlated with the serum collected in same time and the sequential changes in progesterone concentrations in the feces are paralleled in the serum. Therefore, this technique may be suitable for monitoring the estrus cycle in toy poodles.