Measurement Of Progesterone Concentrations Research Articles

APPLICATION OF OVASCAN AND PROGESTERONE CONCENTRATION FOR PREDICTION OF OVULATION DURING ESTRUS IN THE MARE

The estrous cycle of 8 Arabian mares was followed for one year, by visual observation of the estrous behaviour and by teasing with a stallion. Serum progesterone concentration was determined by RIA, Vaginal electric resistence was studied with aid of Ovascan, comapared with the changes in the reproductive tract as determined by rectal examination for prediction of ovulation. The changes in the reproductive tract and the behavioural manifestations expressed during the cycle indicated that the experimental mares were exhibiting estrous year arround, however, 75% of the cycles were observed during the months from March to August and 25% during the. rest of the year. The total number of cycles exhibited by the mares averaged 68 cycles, of these 22 cycles were observed during Summer, 20 during Spring, decreased to 13 cycles during Fall and 13 during Winter. the length of the estrous cycle ranged between 24-26. 31 days, the average estrus length ranged between 5.5-8.6 days andthe luteal phase ranged from 19.04-24.87 days. The ovascan readings showed decline by 50 degrees, constantly, a day preceeding ovulation, compared to the readings during the rest of estrus days and this finding was confirmed by rectal palpation of the reproductive tract in succeeding day which disclosed the disappearance of the mature follicle and the: presence of the ovulation site on the ovary, consequently, the estrous signs faded away gradually within 24-48 hours of the ovuletion. Serum progesterone concentration, during’ the estrus and one day before ovulation ranged between 0.58 -1.05 ng/ml, while mean luteal phase concentration was 10.2820.89 ng/ml. The results of the present study disclosed that the mares in Iraq could be bred any time of the year and it is most probable that measurement of progesterone concentration will have more practical application for monitring ovarian activity and estrus prediction, if. a rapid system of measurment made avilable (as progesterone strip). The application of Ovascan proved effecient tool for prediction of ovulation in the mare, providing information for synchroniztion of service with ovulation, a mean augmenting fertility and avoiding rectal palpation which hinder service.

Reproductive neuroendocrine defects programmed by prenatal testosterone treatment between gestational days 60-90 are amplified by postnatal obesity in sheep.

Polycystic ovary syndrome (PCOS) is the leading cause of anovulatory infertility in women of reproductive age, and obesity can increase the severity and development of the PCOS phenotype. Prenatal testosterone (T) treatment between gestational days 30-90 advanced puberty and disrupted the reproductive and metabolic phenotype in female sheep, recapitulating attributes of women with PCOS, with postnatal obesity amplifying its severity. On the other hand, prenatal T treatment from gestational days 60-90 led to a much milder phenotype. We hypothesized that reproductive neuroendocrine defects programmed by prenatal T treatment between gestational days 60-90 are amplified by postnatal obesity in sheep. Suffolk ewes received T propionate (T; 100mg) or corn oil (C; vehicle) twice weekly from gestational days 60-90. At 5months of age, T lambs were assigned to either a maintenance (100% of NRC requirements) or overfed (130% NRC) diet and C lambs were fed the maintenance diet. We compared the timing of puberty (n = 15/group) determined by twice weekly measurement of progesterone concentrations, estradiol positive feedback responsiveness (n = 8/group) determined by assessing LH secretion in response to exogenous estradiol, periovulatory LH dynamics during the second breeding season (n = 8/group) following synchronization with two injections of PGF2α, and progesterone negative feedback (n = 8/group) determined by characterizing LH pulses during the mid-luteal phase between C, T-maintenance and T-overfed groups. Our findings indicate that postnatal obesity: 1) exacerbated reproductive defects and further deteriorated reproductive cyclicity during the second breeding season (adulthood); 2) did not amplify the impairment in estradiol positive feedback in delaying the timing and amplitude of the LH surge, although it reduced the total amount of LH secreted during the preovulatory LH surge; 3) amplified the reduced responsiveness to progesterone negative feedback manifested as an increase in LH pulse amplitude and peak. These observations, in addition to supporting our previous findings that prenatal T treatment results in reproductive neuroendocrine dysfunction and periovulatory disruptions, provide evidence that these neuroendocrine defects programmed between gestational days 60-90 are amplified by postnatal obesity in female sheep.

Catamenial epilepsy occurence and patterns in a mixed population of women with epilepsy.

We aimed to evaluate the occurrence and distribution of patterns of catamenial epilepsy in a heterogenous cohort of women with epilepsy on no hormonal therapies, enrolled in a prospective, observational study. The primary aim of the study was pregnancy rate in women with epilepsy with no prior reproductive problems. In this analysis, we included women who recorded ≥one menstrual cycle with ≥one seizure. We measured progesterone concentrations for 1-3 cycles. We defined catamenial patterns as ≥two-fold average daily seizure frequency around menstruation (C1), ovulation (C2) and, for anovulatory cycles, from mid-cycle through menstruation (C3). Twenty-three of the 89 enrolled women with epilepsy were eligible for this analysis: 12/23 met criteria for catamenial epilepsy, and 5/23 demonstrated only a C1 pattern, 2/23 only a C2 pattern, 5/23 a combined C1/C2 pattern, and 0/1 women with anovulatory cycles demonstrated a C3 pattern. There were no differences in likelihood of demonstrating a catamenial pattern between those who reported a prior catamenial pattern and those who did not (p=0.855). This analysis demonstrates the utility of app-based tracking to determine a catamenial pattern. Larger prospective studies could confirm these findings and inform potential therapeutic trial designs for catamenial epilepsy.

Viperin (RSAD2) gene expression in peripheral blood mononuclear cells of pregnant crossbred beef cows is altered by Bos indicus genetics

The expression of interferon (IFN) stimulated genes (ISGs) in lymphocytes has been used for pregnancy diagnosis in cattle. However, among-cow variability has yielded sub-optimal predictive accuracy. We hypothesized that the expression of ISGs (ISG15, OAS1, RSAD2, CLEC3B, and AKR1B1) in early pregnancy varies according to the proportion of Bos indicus (B. indicus) genetics on females. Multiparous cows were classified in three genetic groups, High Angus (HA; n = 45 [0–33% Brahman influence]), Angus-Brahman (AB; n = 30 [34–67%]), and High Brahman (HB; n = 19 [68–100%]) and submitted to a Select-Synch + CIDR protocol. Cows that displayed estrus (n = 94) were artificially inseminated (Day0; D0). On D19, blood samples were collected to obtain peripheral blood mononuclear cells (PBMC) and measure progesterone (P4) concentrations. On D30, pregnancy diagnosis was performed. The expression of RSAD2 in PBMC of pregnant cows was positively related to the proportion of B. indicus genetics of the groups, but not the expression of ISG15 and OAS1. In pregnant cows, the proportion of B. indicus genetics was negatively associated to circulating levels of P4 concentrations. The P4 concentrations were related positively with RSAD2 expression. ROC curve results determined that for cattle with B. indicus genetics lower than 67%, the CLEC3B and AKR1B1 combination was the most accurate option to predict the outcome of pregnancy. In cows with more than 68% of B. indicus genetics, RSAD2 provided the best accuracy. In conclusion, there is a relationship between the proportion of B. indicus genetics and the ISGs gene expression in PBMC during pregnancy.

Evaluation of surface plasmon field-enhanced fluorescence spectroscopy for rapid measurement of progesterone concentration in bitches.

To compare progesterone (P4) concentrations measured with surface plasmon field-enhanced fluorescence spectroscopy (SPFS) and chemiluminescence immunoassay (CLIA) in serum and plasma samples of client-owned bitches of various ages and breeds and to determine reference ranges for P4 concentrations at various stages of the estrous cycle. 102 serum samples and 104 plasma samples. In experiment 1, 1 aliquot each of serum and plasma was analyzed for P4 concentration by use of SPFS incorporated in a veterinary-specific point-of-care immunologic analyzer and CLIA. In experiment 2, serum collected from bitches in various stages of the estrous cycle was analyzed for P4 concentration by use of SPFS to establish reference ranges for each stage. In experiment 1, P4 concentrations measured by SPFS and CLIA were highly correlated (serum, r = 0.966; plasma, r = 0.968). In experiment 2, ranges of serum basal (proestrous) P4 concentrations (n = 114) and P4 concentrations at the estimated time of ovulation (76), during pregnancy or diestrus (107), and during the prepartum period (50) measured with SPFS were 0.42 to 1.46 ng/mL, 3.69 to 7.85 ng/mL, 11.73 to 28.24 ng/mL, and 1.54 to 3.22 ng/mL, respectively. Because serum and plasma P4 concentrations measured with SPFS were highly correlated with those measured with CLIA and ranges of serum P4 concentrations measured with SPFS for each of phase of the estrous cycle were well-defined for the large sample size, veterinarians may be able to accurately use this veterinary-specific point-of-care immunologic analyzer with SPFS methodology to determine P4 concentrations of bitches in their daily practice.

Effects of parity, blood progesterone, and non-steroidal anti-inflammatory treatment on the dynamics of the uterine microbiota of healthy postpartum dairy cows.

This study evaluated the effects of treatment with meloxicam (a non-steroidal anti-inflammatory drug), parity, and blood progesterone concentration on the dynamics of the uterine microbiota of 16 clinically healthy postpartum dairy cows. Seven primiparous and 9 multiparous postpartum Holstein cows either received meloxicam (0.5 mg/kg SC, n = 7 cows) once daily for 4 days (10 to 13 days in milk (DIM)) or were untreated (n = 9 cows). Endometrial cytology samples were collected by cytobrush at 10, 21, and 35 DIM, from which the microbiota analysis was conducted using 16S rRNA gene sequence analysis. A radioimmunoassay was used to measure progesterone concentration in blood serum samples at 35 DIM and cows were classified as ˃ 1 ng/mL (n = 10) or ≤ 1 ng/mL (n = 6). Alpha diversity for bacterial genera (Chao1, Shannon-Weiner, and Camargo’s evenness indices) were not affected by DIM, meloxicam treatment, parity, or progesterone category. For beta diversity (genera level), principal coordinate analysis (Bray-Curtis) showed differences in microbiota between parity groups. At the phylum level, the relative abundance of Actinobacteria was greater in primiparous than multiparous cows. At the genus level, there was lesser relative abundance of Bifidobacterium, Lactobacillus, Neisseriaceae, Paracoccus, Staphylococcus, and Streptococcus and greater relative abundance of Bacillus and Fusobacterium in primiparous than multiparous cows. Bray-Curtis dissimilarity did not differ by DIM at sampling, meloxicam treatment, or progesterone category at 35 DIM. In conclusion, uterine bacterial composition was not different at 10, 21, or 35 DIM, and meloxicam treatment or progesterone category did not affect the uterine microbiota in clinically healthy postpartum dairy cows. Primiparous cows presented a different composition of uterine bacteria than multiparous cows. The differences in microbiota associated with parity might be attributable to changes that occur consequent to the first calving, but this hypothesis should be investigated further.

LC-MS/MS quantification of progesterone in ovariectomized miniature swine and its application for preliminary pharmacokinetic study of a novel vaginal sustained release system

Objective We here established a simple, fast, robust and sensitive LC-MS/MS method and validated as well for the quantitative analysis of progesterone (PGT) in ovariectomized (OVX) miniature swine plasma. The method was successfully applied to characterize the pharmacokinetics of a progesterone vaginal drug delivery system. Methods Megestrol acetate was utilized as an internal standard (IS). The separation and detection of PGT from endogenous interference was performed successfully by liquid chromatography with gradient elution and mass spectrometry equipped with positive ESI source using MRM mode. The EVA intravaginal rings (IVRs) were manufactured by hot-melt extrusion (HME), afterward were administrated vaginally to OVX minipigs to evaluate PK study. Results The calibration curve for swine plasma samples across the concentration ranged between 0.25 ng/mL and 100 ng/mL. The intra- and inter-assay accuracy and precision were lower than ±5% and 5.88%, respectively. Recoveries of PGT and IS were ranging from 114–119% and 96.5–112%, respectively. In vitro study showed that the EVA IVRs release 18 mg/day of PGT continuously over 7 days, and corresponding mean PGT plasma concentration in OVX minipigs (CAVG) was 4.892 ng/mL. Conclusion All the study produced reliable results for the measurement of PGT concentration in miniature swine plasma and the method was successfully applied to a PK study for PGT vaginal ring in miniature pigs, which may lay the foundation for further research on the progesterone preparations intended for in assisted reproductive technology.

Administration of nerve growth factor-β to heifers with a pre-ovulatory follicle enhanced luteal formation and function and promoted LH release

The objective of this study was to determine the effects of bovine nerve growth factor-β (NGF) on pre-ovulatory follicle vascular area, LH release, ovulation, and luteal function when administered systemically to heifers. Post-pubertal Holstein heifers (n = 12) received an intravaginal progesterone-releasing device (CIDR) and GnRH agonist (100 μg IM). The CIDR was removed 5 d later, and heifers were given dinoprost (25 mg IM) at CIDR removal and 24 h later, followed by a second dose of GnRH agonist 48 h later. Heifers were randomly assigned to treatments using a cross-over design. For example, heifers assigned to NGF (250 μg reconstituted in 12 mL PBS IM) in replicate 1 were assigned to control (12 mL PBS IM) in replicate 2. Transrectal ultrasonography was performed before treatment and repeated every 4 h up to 32 h to determine the pre-ovulatory follicle diameter, vascular area, and ovulation. Serum samples were obtained to assess LH concentrations during the periovulatory period and every 2 d post-ovulation for measuring progesterone concentrations. A subset of heifers had luteal biopsies performed on days 9 (n = 6 per treatment) and 14 (n = 6 per treatment) post-ovulation to count luteal cell numbers and measure relative mRNA abundance for steroidogenic and angiogenic enzymes and LH receptor. Treatment with NGF increased pre-ovulatory follicle diameter (P = 0.02) and serum LH concentrations (P = 0.03) but did not affect time to ovulation (P = 0.42). Heifers treated with NGF had increased serum progesterone concentrations in the subsequent luteal phase (P = 0.03), but no change in vascular area of the follicle (P = 0.16) or CL (P = 0.20). Heifers treated with NGF had a greater number of small luteal cells (P < 0.01) and a tendency for increased LH receptor (LHR) mRNA abundance in the CL (P = 0.10). There was also increased steroidogenic acute regulatory protein (STAR; P = 0.05) and a tendency for increased cytochrome P450 family 11 (CYP11A1; P = 0.10) mRNA abundance in the CL of NGF-treated heifers. There was decreased prostaglandin E2 synthase (PGES; P = 0.03) and its receptor (PGER; P = 0.05) mRNA abundance and a tendency for decreased cytochrome P450 family 17 subfamily A member 1 (CYP17A1; P = 0.08) and hydroxysteroid 17-beta dehydrogenase (HSD17B; P = 0.06) mRNA abundance in the CL of NGF-treated heifers. Administration of NGF improved CL function in heifers potentially as a result of increased LH release.

A shortened resynchronization treatment for dairy cows after a nonpregnancy diagnosis

We hypothesized that a shortened version of a modified Ovsynch program (OVS: GnRH-1 − 7 d − PGF2α-1 − 24 h − PGF2α-2 − 32 h − GnRH-2 − 16 h − AI) that excluded GnRH-1 to resynchronize ovulation in cows bearing a corpus luteum (CL) after a non-pregnancy diagnosis (NPD) or including progesterone supplementation with the OVS treatment for cows without a CL would produce shorter inter-insemination intervals and pregnancy per AI (P/AI) not different from that of cows treated with the OVS treatment. Of the 1697 lactating Holstein cows enrolled in this experiment, complete data were available for only 1584 cows because the remainder was not treated, inseminated per treatment design, left the herd before pregnancy diagnosis, or some other outcome was missing. Cows were enrolled in the study and assigned to either of three treatments at NPD (32 ± 3 d after AI [Day 0]). Cows with a detected CL were assigned randomly to: (1) a modified Ovsynch (OVS; GnRH-1 − 7 d − PGF2α-1 − 24 h − PGF2α-2 − 32 h − GnRH-2 − 16 h − AI) or (2) Short Synch (SS; PGF2α-1 − 24 h − PGF2α-2 − 32 h − GnRH-2 − 16 h − AI). Cows with no CL were assigned to OVS plus a progesterone insert (CIDR). Blood was collected at NPD to measure progesterone concentration and determine accuracy of treatment assignment (progesterone ≥ 1 ng/mL for a functional CL). Overall progesterone concentration at NPD was less in OVS + CIDR cows (1.5 ± 0.3 ng/mL) than in OVS (5.2 ± 0.2 ng/mL) or SS cows (3.7 ± 0.3 ng/mL). No differences in luteolytic risk (progesterone < 0.5 ng/mL at 72 h after PGF2α-1) were detected after PGF2α (>96.7%) and ovulation risk after GnRH-2 was 93.8, 91.7, and 86.2% for SS, OVS, and OVS + CIDR, respectively. Mean and median inter-insemination interval were less in SS (mean = 34.3 ± 0.05 d [median = 35 d] than OVS cows (40.2 ± 0.05 d [42 d]), but that in OVS cows did not differ from OVS + CIDR cows (41.4 ± 0.05 d [42 d]). Technicians were more accurate in visually detecting a functional CL than a non-functional CL (81.2 vs. 61.1%). Sensitivity of detecting a functional CL by technicians averaged 91.2%, but specificity was 39.8%. Pregnancy per AI at 32 ± 3 d after AI was less for SS (16.5% [n = 115]) than OVS (29.3% [n = 133] when a functional CL was inaccurately detected, but did not differ when a functional CL was detected accurately (27.6% [n = 561] vs 30.3% [508]). Pregnancy per AI did not differ between OVS and OVS + CIDR cows regardless of CL status. Short synch is an alternative to the entire modified Ovsynch program to produce similar P/AI when the CL status was detected accurately, and regardless of functional CL status, SS reduced inter-insemination intervals by 7 d.

PERBANDINGAN KONSENTRASI PROGESTERON SELAMA SIKLUS BIRAHI PADA DOMBA WARINGIN YANG DIINDUKSI PGF2α DAN KOMBINASI PGF2α DAN GnRH

This study aimed to compare the progesterone concentrations in Waringin sheep estrus cycle which induced by PGF2α or the combination of PGF2α and GnRH. This study used six Waringin sheep those were divided into twice groups, K1 and K2. K1(n=3) were injected by 7.5 mg PGF2α intramuscularly for 10 days, while K2 (n = 3) were injected by PGF2α and GnRH. On the first day K2 were injected by 7.5 mg PGF2α, then followed by injection 50 µg of GnRH on day 8th and re-injected by 7.5 mg PGF2α on day 15th. On day 18th, sample was re-injected using 50 µg GnRH. Waringin sheep those showed estrus symptoms were detected visually and with signs showed by sheep stud. Blood samples were taken on the 7th, 14th, and 21st day after the peak heat. The measurement of progesterone concentration was conducted by enzyme linked immunosorbent assay (ELISA). The results showed those the concentrations of progesterone on day 7th, 14th, and 21st on K1 vs K2 were 1.324±1.079, 7.607±8.922, and 5.220±1.653 vs 4.705±3.369, 4.184±5.512, and 1.797±0.898 ng/mL (P&gt;0,05), respectively. In conclusion, the concentration of progesterone of Waringin sheep after inducing with PGF2α or combination PGF2α and GnRH at different cycle periode did not show differences. Keywords: Estrus Cycle, GnRH, PGF2α, Progesterone, Waringin Sheep

Validation of a blubber-based endocrine pregnancy test for humpback whales.

Baleen whales have few identifiable external indicators of pregnancy state, making it challenging to study essential aspects of their biology and population dynamics. Pregnancy status in other marine mammals has been determined by measuring progesterone concentrations from a variety of sample matrices, but logistical constraints have limited such studies in free-swimming baleen whales. We use an extensive blubber sample archive and associated calving history data to retrospectively identify samples that correspond to pregnant females and develop a progesterone-based pregnancy test for humpback whales. The lowest pregnant blubber progesterone concentration was 54.97 ng g-1, and the mean for the known-pregnant group was 198.74 ± 180.65 ng g-1. Conversely, females known to be below the minimum age of sexual maturity (juvenile females) had an overall low mean progesterone concentration (0.59 ± 0.25 ng g-1), well below the known-pregnant range. Of the mature females that did not return with a calf (n = 11), three fell within the known-pregnant range (320.79 ± 209.34 ng g-1), while the levels for the remaining eight were two orders of magnitude below the lowest known-pregnant level (1.63 ± 1.15 ng g-1). The proportion of females that did not return with a calf but had values similar to known-pregnant females are consistent with rates of calf mortality, but other potential explanations were considered. Our findings support a validated blubber endocrine assignment of pregnancy corroborated with field life history information, a first for any baleen whale species. The progesterone values we measured were similar to those found in different pregnancy states of other cetaceans and support using blubber biopsy samples for assigning pregnancy in humpback whales. This method can be applied to existing archives or new samples to better study life history and population demography broadly across species and populations.

Initial validation of blubber cortisol and progesterone as indicators of stress response and maturity in an otariid; the California sea lion (Zalophus californianus)

Chronic stress can have detrimental effects on an individual’s health and reproductive success. The use of cortisol quantification as an indicator of stress in free-ranging cetaceans and phocids is increasing but no studies have applied this technique on blubber in otariids. We measured cortisol concentrations in blubber samples obtained from California sea lions, Zalophus californianus, stranded in San Diego County and those incidentally killed in the California drift gillnet fishery. We also measured progesterone concentrations to assess female reproductive status and, in males, as a potential secondary measure of adrenal steroid production. Blubber cortisol and progesterone values were compared across demographic groups (sex and maturity), season, and proportion blubber lipid extracted. Stranded animals (247.3±70.767SEng/gblubber) had significantly higher cortisol concentrations compared to fishery bycaught (8.1±2.108SEng/gblubber) animals. These findings are likely driven by inherent differences in the cause of death and associated nutritional state coupled with the mean duration of expiration for these two groups of animals (i.e., the duration from an animal’s initial perception of the threat-to-self until death). The duration of transition from healthy state to death in stranded animals is on the order of many hours to weeks while in fishery bycaught animals, this transition occurs much more rapidly (i.e., seconds to tens of minutes). The presumed longer duration of the mortality event in stranded animals gives sufficient time for elevated cortisol to diffuse into the blubber. No significant differences between demographic groups, or season were found. However, blubber cortisol declined inversely with proportion blubber lipid extracted, suggesting utility in assessing long-term nutritional status. Blubber progesterone was significantly higher in mature females than immature females (153.8±54.546SEng/gblubber and 9.7±3.60SEng/gblubber respectively), containing on average 15 times more progesterone, irrespective of pregnancy state. Additionally, a significant relationship between mean cortisol and progesterone was found in males with >35% blubber lipid (p<0.0001). This study is an initial step in validating blubber cortisol and progesterone concentrations as a potential marker of stress response and reproductive state, respectively, in otariids. Especially when paired with dart biopsying, this approach could represent a relatively rapid way to assess baseline stress, nutritional status and reproductive states in otariids while minimizing the effects of sampling.

Luteal activity and effect of dietary energy restriction on follicular development in lactating cows.

The aim of this research has been to evaluate the presence of anomalies in the ovarian cycle activity during postpartum and to verify whether 72-hr dietary fasting during the dominance phase, the phase before ovulation, might modify the ovarian follicle population. The presence of anomalies in ovarian cycle activity has been evaluated in 30 Italian Friesian cows starting from 20days postpartum until 211days of lactation. Long oestrus and brief dioestrus or scarce luteal activity have been the main anomalies found through measuring progesterone concentrations in the whey. Until 100days of lactation, the BCS values of the problematic animals have been significantly lower than those in animals with normal ovarian activity. After 100days of lactation, the ovarian anomalies continued to appear despite the fact that all the animals have reached comparable BCS values. Starting from the results of this trial, the effect of 72-hr dietary fasting on dominant follicles has been studied in six cows. Ultrasonography revealed that the diameter of the follicles at 71days postpartum has been significantly lower than at 181days. A 72-hr dietary restriction at 101 and 211days postpartum did not affect the size of the dominant follicle. However, at 101days postpartum, half of the animals presented follicular cysts. The effect of fasting differed if the animal has been in early postpartum or 211days of lactation. Further researches are necessary to understand how different metabolic conditions can modify the follicular population but on the other hand the study shows the utility for farmers and field veterinarians of monitoring the resumption of the ovarian cycle postpartum through the whey progesterone concentrations.

Effectiveness of Heat Detection Patches for Determining Cyclicity Status in Peripubertal Heifers

Response to most estrous synchronization programs is improved in cycling females, compared to pre-pubertal heifers or anestrus cows. Cycling status is often monitored for research purposes by measuring progesterone concentrations in serum. Producers may want to use information on cyclicity status when determining which estrous synchronization program to use or as a component of their heifer selection. Heat detection patches are routinely used to aid or reduce time needed for heat detection. The most data on their use is for a 3 to 5-day period after estrous synchronization. Little information is available on retention of patches for longer time periods or how they may compare with serum progesterone for determining cyclicity status. Therefore, the objective of the current study was to compare Standing Heat and Estrotect heat detection patches with serum concentrations of progesterone for identifying cycling (pubertal) heifers before the start of the breeding season.

Comparison of three devices for the automated detection of estrus in dairy cows

Considerable technological advances have been made in the automated detection of estrus in dairy cattle, but few studies have evaluated their relative performance on the same animals or assessed cow-related factors that affect their performance. Our objective was to assess the performance and reliability of three devices commercially available in France for cow estrus detection. The devices were a pedometer (PM; Afitag) and two activity meters (AM1; Heatime-RuminAct, and AM2; HeatPhone). Two algorithms were tested for AM2. We fitted 63 lactating Holstein cows with the three detectors from calving to 90 days after calving. The onset and pattern of cyclicity were monitored from 7 to 90 days postpartum measuring progesterone concentration in milk twice weekly. A total of 211 ovulations were identified. Cyclicity was classified as normal in 60% of cows (38/63). Calculated over the operating period of all the devices (179 periods of estrus), the sensitivities and positive predictive values were, respectively, 71% and 71% for PM, 62% and 84% for AM1, 61% and 67% for the first algorithm of AM2, and 62% and 87% for the second algorithm of AM2. Both activity meters had a lower sensitivity but a higher positive predictive value than the PM (P < 0.05). For all devices, the performance in estrus detection was much poorer at the first postpartum ovulation than at subsequent ovulations (P < 0.05). Lactation rank and milk production affected some devices (P < 0.05). These devices could be used to reinforce visual observations, especially after 50 days postpartum, the minimum recommended delay to insemination. However, their full benefit remains to be verified in different farming systems and taking into account the specific objectives of the dairy farmer.

Detection of Glycosylated Proteins in Rabbit Oviductal Isthmus and Uterine Endometrium During Early Embryo Development

In this study, lectin histochemistry was performed on paraffin sections to compare carbohydrate expression of oviductal isthmus and uterine endometrium in rabbits during early embryo development. Rabbit embryos are surrounded not only by the zona pellucida but also by tubal secretion-derived mucinous coat material, the mucin coat. Twenty sexually mature females were euthanized at 0 (pre-ovulatory group) and 24, 72 and 96h after insemination (pseudopregnancy group). The following lectin-binding agents were used: Arachis hypogaea, Peanut (PNA) to label galactosyl (β-1,3)N- acetyl-galactosamine, Dolichos biflorus Agglutinin (DBA) to label galactosyl (β-1,4)N- acetyl-galactosamine, Lens curinaris (LCA) to label α--mannose, α-d-glucose and Pisum sativum agglutinin (PSA) to label α-d-mannose, α-d-glucose. Blood was collected by cardiac puncture, and direct enzyme immunoassay technique was used to measure progesterone concentration. A significant increase in total plasma progesterone concentrations was detected at 96h post-ovulation when compared with 0, 24 and 72h post-ovulation (2.9±0.5 vs 0.5±0.15, 1.6±0.5 and 1.5±0.4ng/ml, at 96h vs 0, 24 and 72h post-ovulation, respectively). No differences between pre-ovulatory and pseudopregnant females were observed for glycoprotein localization in isthmus. In contrast, in the endometrium, differences in the glycoprotein detection between pre-ovulatory and pseudopregnant stages were detected. PNA to label galactosyl (β-1,3)N- acetyl-galactosamine was not detected at the pre-ovulatory stage, but its presence was detected at 24h after ovulation. Both PSA and LCA to label α-d-mannose, α-d-glucose were only detected at 72h after ovulation. DBA detection was similar for all stages of the reproductive cycle. Therefore, N-acetyl-galactosamine secreted from isthmus could be involved in the formation of the embryonic mucin coat. d-galactose (PNA), d-glucose and d-mannose (PSA and LCA) might be crucial for the implantation period.

Hormonal predictors of sexual motivation in natural menstrual cycles

Little is known regarding which hormonal signals may best predict within- and between-women variance in sexual motivation among naturally cycling women. To address this, we collected daily saliva samples across 1–2 menstrual cycles from a sample of young women; assayed samples for estradiol, progesterone, and testosterone; and also collected daily diary reports of women's sexual behavior and subjective sexual desire. With respect to within-cycle, day-to-day fluctuations in subjective desire, we found evidence for positive effects of estradiol and negative effects of progesterone. Desire exhibited a mid-cycle peak, similar to previous findings; measured progesterone concentrations statistically mediated the fall in desire from mid-cycle to the luteal phase, but no combination of hormone measures substantially mediated the follicular phase rise in desire, which suggests that other signals may be implicated in this effect. Hormonal predictors of within-cycle fluctuations in sexual behavior generally reached only trend levels of statistical significance, though the patterns again suggested positive effects of estradiol and negative effects of progesterone. Between-women and within-women, between-cycle effects of hormone concentrations were generally absent, although statistical power was more limited at these higher levels of analysis. There were no significant effects of testosterone concentrations when controlling for the effects of estradiol and progesterone, which raises questions regarding the importance of this hormone for the regulation of sexual motivation in natural cycles. Our study is among the first to identify hormonal predictors of within-cycle fluctuations in sexual motivation, and thus adds novel evidence regarding the endocrine correlates of human sexuality.

Progesterone Profiles in the Caudal Vena Cava and Jugular Vein in Response to Pulsatile Luteinizing Hormone Stimulation Induced by GnRH Treatment During the Mid-luteal Phase in Lactating Dairy Cows

The aim of this study was to examine whether increased frequency of luteinizing hormone (LH) pulses influences luteal progesterone (P4) secretion by measuring progesterone concentrations at the secreted (caudal vena cava) and circulating levels (jugular vein) in lactating dairy cows. Cows received six intravenous administrations of 2.5 μg of GnRH (gonadorelin acetate, n=4) or 2 ml saline (n=3) at 1-h intervals on 12.4 ± 0.4 (mean ± SE) days after ovulation. Blood samples were collected from the caudal vena cava and jugular vein every 12 min for 12 h (6 h before and after treatment). During the 6 h after treatment, frequency of LH pulses (5.3 ± 0.3 and 3.0 ± 0.0 pulses/6 h) and mean LH concentration (0.50 ± 0.06 and 0.38 ± 0.05 ng/ml) were greater (P<0.05) in GnRH-treated cows than in saline-treated cows. Mean P4 concentration and amplitude of P4 pulses in the caudal vena cava during the 6 h after treatment were greater (P<0.05) in GnRH-treated cows than in saline-treated cows, but the frequency of P4 pulses was not different between the groups. Mean P4 concentration in the jugular vein during the 6 h after treatment was also higher (P<0.05) in GnRH-treated cows than in saline-treated cows (7.0 ± 1.3 and 5.4 ± 0.9 ng/ml). These results indicate that the increased frequency of LH pulses stimulates progesterone secretion from the functional corpus luteum and brings about higher P4 concentrations in the circulating blood in lactating dairy cows.

The Localization of Toll‐like Receptor 2 (TLR2) in the Endometrium and the Cervix of Dogs at Different Stages of the Oestrous Cycle and with Pyometra

The aim of this study was to localize and evaluate the role of Toll-like receptor 2 (TLR2) in the endometrium and cervix of bitches at different stages of the oestrous cycle and in bitches with pyometra. Sixty-seven nulliparous dogs, ranging in age from 1 to 13 years, were allocated amongst five groups (pro-oestrus; n = 7, oestrus; n = 10, dioestrus; n = 16, anoestrus; n = 11, pyometra; n = 23). Blood samples were collected for the measurement of progesterone concentration. The mean progesterone concentration was analysed as a parameter for validating the stage of the oestrous cycle in bitches. Tissues collected from uterine horn and cervix were fixed in 4% paraformaldehyde for immunohistochemical examination of TLR2. The expression of TLR2 was assessed semi-quantitatively. No pathological changes were found in the uterine samples of healthy dogs. In bitches with pyometra, the glandular epithelium expressed TLR2 more intensely than the surface epithelium. The expression of TLR2 in the glandular epithelium was also significantly higher in healthy dogs at oestrus, dioestrus and dogs with pyometra compared with anoestrous dogs (p < 0.01). The expression of TLR2 in the stroma was not observed in the group of healthy dogs at all stages. The surface epithelium of cervix in dogs with pyometra expressed TLR2 significantly more intensely than did the stoma, whereas the expression of TLR2 during oestrus and dioestrus was absent in the stroma of cervix. This study provides the first report of immunohistochemical localization of TLR2 in the canine reproductive tract. In the present study, TLR2 was expressed in endometrial epithelium but was absent in the endometrial stroma of healthy dogs at all oestrous cycle stages. These findings suggest differential expression of TLR in endometrial cells. On the other hand, the lack of TLR2 in the stroma of healthy uteri of dogs may predispose to infection from the invading pathogens once the epithelial cells have been destroyed by the pathogens, especially Gram-positive bacteria.

Association of milk yield and body condition score indices with the commencement of luteal activity after parturition in high producing dairy cows

Summary The aim of the present study was to investigate the relation between different indices of milk yield and body condition score (BCS) with the commencement of the luteal activity (C-LA) during the postpartum period in high producing dairy cows. Seventy-one multiparous healthy (free of detectable reproductive disorders) Holstein dairy cows (mean peak milk yield = 56.7 ± 7.4 kg) were used in the present study. Transrectal ultrasound scanning was performed twice weekly from the 1st to the 8th week postpartum. Blood samples were also collected twice weekly to measure serum progesterone (P4) concentrations. The BCS was monitored weekly after calving. Cows with serum P4 concentrations ≥1 ng/ml on at least 2 consecutive blood samplings were considered to have commenced luteal activity. The C-LA was observed in 51 out of 71 cows (71.8%) earlier than 45 days postpartum, while 20 out of 71 cows (28.2%) showed the C-LA later than 45 days postpartum. Among the indices defined for the milk yield pattern in the present study, difference in milk yield between the 1st week and the peak week and the peak of milk yield were significantly (P≤0.05) lower in cows that commenced their luteal activity earlier than 45 days postpartum. Cows losing ≥1 unit BCS at the 3rd week postpartum showed their C-LA significantly later than those cows losing less BCS during the same interval (P≤0.05). In conclusion, among the different indices defined for the milk yield and BCS in the present study, the higher milk yield at the peak and lower A-BCS (Area under the chart of BCS change) were the main factors associated with delayed commencement of luteal activity in clinically healthy high producing dairy cows.