Urinary Progesterone Metabolites Research Articles

Progesterone monitoring and gestation length in Amazonian manatees (Trichechus inunguis)

Knowledge of pregnancy length and the physiological parameters during pregnancy is an important tool for pregnancy diagnosis in free-ranging and captive individuals and for the correct husbandry of captive pregnant females. Therefore, this study aimed to describe the progesterone pattern during pregnancy in T. inunguis and to estimate the pregnancy length in this species. Blood or urine samples from a total of eight pregnancies were collected from six captive female Amazonian manatees. Serum progesterone and urinary progesterone metabolites were measured by radioimmunoassay and enzyme immunoassay. The mean of serum progesterone throughout pregnancy was 5.17 ± 4.71 ng/ml (0.08–19.71 ng/ml) and the mean urinary progesterone metabolites concentration was 11.34 ± 12.28 ng/mg Cr (1.00 – 54.71 ng/mg Cr). A pregnancy length of approximately 12–13 months was observed (369.6 ± 27.2 days, range 352–401 days). The pregnancy length observed in this study falls within the range reported for other sirenians. During the first four months, there was a high level of progesterone. After that, the hormone levels began to decrease. Low progesterone levels during most of pregnancy were also observed in other sirenians. This hormonal pattern suggests the existence of the paracrine action of progesterone or the role of other hormones in the maintenance of pregnancy. However, further studies are necessary to clarify the pregnancy endocrinology in Amazonian manatees.

The Predictive Value of Urinary Progesterone Metabolite PdG Testing in Pregnancy Outcomes

Background: Progesterone is essential for establishing and maintaining a pregnancy. However, premature atresia of the corpus luteum (CL) or sub-optimal progesterone secretion after ovulation may result in the decreased ability to establish or maintain a pregnancy. The current limitations of serum hormone testing fail to give a complete picture of progesterone production across the luteal phase required for pregnancy.

Fecal progesterone and estrogen metabolite monitoring for cyclicity and pregnancy in southern tamandua (Tamandua tetradactyla).

Reproductive management of the southern tamandua (Tamandua tetradactyla) should include timed introductions for breeding to minimize aggression and pregnancy monitoring. Since serial blood sampling could cause unnecessary stress, and urinary progesterone metabolites are found in very low concentrations, this study sought to validate progesterone and estradiol enzyme immunoassays for measuring fecal progesterone metabolite (FPM) and fecal estrogen metabolite (FEM) concentrations in two females. Peaks in FEM concentrations coincided with breeding and conception, were 5-6 times higher than baseline concentrations, and were followed by clear luteal phases distinguished by FPM concentrations 5-6 times higher than baseline concentrations. FPM concentrations during the first 30-53 days of gestation overlapped with luteal phase concentrations, thereafter increasing to 8-25 times higher than baseline concentrations. FEM concentrations during the first 41-44 days of gestation remained near basal values for one female, whereas concentrations were 1.8 times higher than baseline for the second. FEM concentrations became elevated for the former by 44 days of gestation and increased further for the latter after 53 days, ultimately averaging four times higher than baseline for both females. The biphasic increase in FPM and FEM concentrations, follicular and luteal phase durations (follicular: 7 ± 1 days, luteal: 25 ± 1 days), total cycle length (41 ± 1 days), and gestation (161-165 days) documented in this study were consistent with previous reports from serum and urine analyses. Monitoring FPM and FEM is a reliable noninvasive method for tracking reproductive cycles and pregnancy in southern tamandua that overcomes the challenges associated with serum or urinary hormone analysis.

Interrelationship among annual cycles of sex steroids, corticosterone and body condition in Nyctibatrachus humayuni

Synergism between extrinsic and intrinsic factors is crucial for the seasonality of reproduction. Environmental factors such as photoperiod and temperature activate the hypothalamus-pituitary-gonadal axis leading to the secretion of steroid hormones that are crucial for reproduction. Sex steroids are not only essential for the maturation of gonads, but also for development of secondary sexual characters in males and reproductive behaviour of both the sexes. In the present study, we quantified the urinary testosterone (UTM) and corticosterone (UCM) metabolites in males and urinary estradiol metabolites (UEM) and UCM in females of Nyctibatrachus humayuni for two consecutive years to determine annual and seasonal variation in the levels of sex steroids, corticosterone and body condition index (BCI). The results show that sex steroids were highest during the breeding season and lowest during the non-breeding season in both the sexes. An increase in UTM and UEM was observed in males and females respectively during the breeding season. Testicular histology showed the presence of all stages of spermatogenesis throughout the year indicating that spermatogenesis is potentially continuous. Ovarian histology showed the presence of vitellogenic follicles only during the breeding season indicating that oogenesis is strictly seasonal. In males, UCM levels were highest during the breeding season, while in females their levels were highest just prior to the breeding season. In males, BCI was highest during the pre-breeding season, declined during the breeding season to increase again during the post-breeding season. In females, BCI was comparable throughout the year. In males, UTM levels were positively correlated with UCM levels but negatively correlated with BCI. Interestingly, UEM, UCM and BCI were not correlated in females. These results indicate that N. humayuni exhibits an associated pattern of reproduction. Quantification of urinary progesterone metabolites (UPM) during the breeding season showed UPM levels were higher in post-spawning females, suggesting the significance of progesterone in ovulation. Further, non-invasive enzyme immunoassay has been successfully standardized in N. humayuni for the quantification of urinary metabolites of steroid hormones.

Reproductive biology of captive female southern hairy-nosed wombats (Lasiorhinus latifrons). Part 2: oestrous behaviour.

The poor captive breeding success of southern hairy-nosed wombats (SHNWs; Lasiorhinus latifrons) has been attributed to the difficulty in accurately characterising oestrous behaviour and their relationship to circulating reproductive hormones. Over two wombat breeding seasons, the use of infrared cameras for 24-h remote behavioural monitoring and the analysis of urine samples collected from seven captive females, were investigated to determine the relationship between behaviour and changes in urinary progesterone metabolites (P4M). Urinary P4M was divided into two concentrations: (1) ≤ baseline P4M values and (2)>baseline P4M values and evaluated against urine volume (mL) and the duration (s) and frequency of 23 behaviours recorded for 8 days surrounding D0 of the luteal phase (D0: a sustained increase in P4M for three or more consecutive days). When P4M was ≤ baseline, the duration of urination and volume both decreased, whereas the duration and frequency of both pacing and rump bites by the female towards the male increased. These results suggest that there were detectable behavioural changes that can be mapped to the changes in the SHNW oestrous cycle, which may be used as behavioural indicators to identify the reproductive status of females.

Tamoxifen for the treatment of breakthrough bleeding with the etonogestrel implant: a randomized controlled trial

ObjectiveThe etonogestrel (ENG) subdermal implant can cause frequent breakthrough bleeding in some users. The objective of this study was to evaluate whether a short course of tamoxifen reduces bleeding/spotting days compared to placebo in ENG implant users. Study designIn this double-blind trial, we randomized ENG implant users with frequent or prolonged bleeding or spotting to tamoxifen 10 mg or placebo twice daily for 7 days, to be started after 3 consecutive days of bleeding/spotting. Treatment was repeated as needed up to three times in 180 days. Subjects completed a daily text message bleeding diary. A sample size of 56 provided 80% power to detect a difference of 6 days of bleeding/spotting per 30 days by two-sample t test. Ovulation was monitored by urinary metabolites of progesterone. ResultsFrom March 2014 to February 2015, 56 women enrolled. Fifty-one completed at least 30 days of follow up, and 34 completed 180 days. Compared to women randomized to placebo, women randomized to tamoxifen reported 5 fewer days of bleeding/spotting over 30 days (95% confidence interval [CI] −9.9 to −0.05, p=.05), and 15.2 more continuous bleeding-free days (95% CI 2.8–27.5 days, p=.02) after first use of study drug. Conclusions could not be drawn after 30 days due to higher-than-expected dropout. No ovulation was detected. ConclusionFirst use of tamoxifen by ENG implant users reduces bleeding/spotting days and provides a longer cessation of bleeding/spotting than placebo, without compromising ovulation suppression. Further study is needed to determine whether this effect is maintained with repeat use. ImplicationsWomen with frequent ENG implant-related breakthrough bleeding may experience a reduction in bleeding/spotting days and an increase in continuous bleeding-free days in the month following first use of tamoxifen. This short course of tamoxifen was well tolerated with bleeding cessation noted within a median of 5 days.

At-Home Detection of Ovulation by Urinary Progesterone

Many in the natural family planning community have looked forward to the development of at-home urinary progesterone test kits, like Ovulation Double Check, by MFB Fertility, because it is an “excellent marker for the closure of the fertile window.” Several research studies have shown that the urinary progesterone metabolite P3G consistently marks rises above baseline more than one day after ovulation. Previously, a progesterone level could be measured only by a serum test, which requires a prescription and blood draw and can be expensive. A urine progesterone metabolite P3G test, which costs about $5, is an attractive alternative.

Detecting evidence of luteal activity by least-squares quantitative basal temperature analysis against urinary progesterone metabolites and the effect of wake-time variability

Objective To assess computerised least-squares analysis of quantitative basal temperature (LS-BT) against urinary pregnanediol glucuronide (PdG) as an indirect measure of ovulation, and to evaluate the stability of LS-QBT to wake-time variation. Study design Cross-sectional study of 40 healthy, normal-weight, regularly menstruating women aged 19–34. Participants recorded basal temperature and collected first void urine daily for one complete menstrual cycle. Evidence of luteal activity (ELA), an indirect ovulation indicator, was assessed using Kassam's PdG algorithm, which identifies a sustained 3-day PdG rise, and the LS-QBT algorithm, by determining whether the temperature curve is significantly biphasic. Cycles were classified as ELA+ or ELA−. We explored the need to pre-screen for wake-time variations by repeating the analysis using: (A) all recorded temperatures, (B) wake-time adjusted temperatures, (C) temperatures within 2 h of average wake-time, and (D) expert reviewed temperatures. Results Relative to PdG, classification of cycles as ELA+ was 35 of 36 for LS-QBT methods A and B, 33 of 34 (method C) and 30 of 31 (method D). Classification of cycles as ELA− was 1 of 4 (methods A and B) and 0 of 3 (methods C and D). Positive predictive value was 92% for methods A–C and 91% for method D. Negative predictive value was 50% for methods A and B and 0% for methods C and D. Overall accuracy was 90% for methods A and B, 89% for method C and 88% for method D. The day of a significant temperature increase by LS-QBT and the first day of a sustained PdG rise were correlated ( r = 0.803, 0.741, 0.651, 0.747 for methods A–D, respectively, all p < 0.001). Conclusion LS-QBT showed excellent detection of ELA+ cycles (sensitivity, positive predictive value) but poor detection of ELA− cycles (specificity, negative predictive value) relative to urinary PdG. Correlations between the methods and overall accuracy were good and similar for all analyses. Findings suggest that LS-QBT is robust to wake-time variability and that expert interpretation is unnecessary. This method shows promise for use as an epidemiological tool to document cyclic progesterone increase. Further validation relative to daily transvaginal ultrasound is required.

ARE CAPE GROUND SQUIRRELS (XERUS INAURIS) INDUCED OR SPONTANEOUS OVULATORS?

Little is known about the ovulation patterns in ground squirrels, even though the timing of ovulation could influence mate-order effects and sperm competition, which have been documented in a number of species. The Cape ground squirrel (Xerus inauris) is a social, nonhibernating, semifossorial rodent with aseasonal reproduction. Evidence suggests that sperm competition is an important aspect of male reproductive success in this species. We examined whether the Cape ground squirrel is an induced or spontaneous ovulator. Fifteen reproductive adult females were removed from natal colonies and housed individually. Females were subjected to 1 of 3 trials: no male contact, chemical and visual contact but no physical contact with intact males, or direct contact with epididymectomized males. Females from each of the 3 experimental treatments exhibited similar urinary progesterone metabolite concentrations and qualitative ovarian histology. Neither the presence of males nor copulation appear necessary for ovulation to occur, and ovulation in this social African ground squirrel appears to be spontaneous.

Defining the Relationship Between Ovarian Hormones and Migraine Headache

(1) To determine whether the attack characteristics of migraine differ between different intervals of the menstrual cycle; (2) To ascertain whether the "rate of change,""magnitude of change," or "total burden" of urinary hormone metabolites correlates with headaches outcome measures during different intervals of the menstrual cycle. The mechanisms through which migraines are influenced by ovarian hormones remain unclear. No previous studies until now have identified "hormonally defined" time intervals within the female menstrual cycle and compared headache outcome measures among these intervals in female migraineurs. Daily headache diary data were obtained from 21 female migraineurs during three native menstrual cycles. Daily urine samples were collected and later assayed for estrogen and progesterone metabolites. Seven 3-day time intervals were identified within each menstrual cycle based on urine hormone measurements. Primary (headache index) and secondary (disability index, headache severity, and headache frequency) outcome measures were compared between intervals using the mixed model approach. "Rates of change,""magnitude of change," and the "total burden" of ovarian hormones were estimated from urine hormone metabolites and correlated with headache outcome measures. The headache index was significantly different across different intervals of the menstrual cycle (P values <.001) and was higher during menstrual intervals (first 6 days of the menstrual cycle) than during mid-cycle and mid-luteal intervals (P < .002). Similarly, secondary outcome measures were highest during the menstrual intervals. "Higher burdens" of urinary progesterone metabolites were positively correlated with headache outcome measures during the luteal intervals of the menstrual cycle. "Rates of change" and the "magnitude of change" of urinary hormone metabolites did not correlate with headache outcome measures. Migraine headache is more severe, disabling, and frequent during the menstrual intervals of the female reproductive cycle than during mid-luteal or mid-cycle intervals. Progesterone metabolites may play a role in modulating migraine headaches during luteal intervals of the menstrual cycle.

Follicular, endocrine and behavioural dynamics of the Indian rhinoceros (Rhinoceros unicornis) oestrous cycle

Longitudinal ultrasound, behaviour and endocrine evaluations were conducted, over 14 to 18 months, in two young female Indian rhinoceroses (Rhinoceros unicornis) to characterize the oestrous cycle. Both females showed the same pattern of follicular development producing a large follicle (10-12 cm diameter) on one of the ovaries that persisted for 8.5 +/- 4.68 days before spontaneously ovulating. Ovulation occurred in all eight cycles monitored in a 6- to 7-year-old female versus 10 out of 14 cycles monitored in a 5- to 6-year-old female. Ultrasound examinations confirmed follicular collapse 48 h following the onset of behavioural oestrus in ovulatory cycles, while anovulatory cycles were associated with the formation of a haemorrhagic follicle. The day of behavioural oestrus corresponded to peak urinary oestrogen conjugate concentrations for each cycle, but anovulatory cycles had lower concentrations on the day of behavioural oestrus compared with ovulatory cycles. A transient increase in urinary progesterone metabolite concentrations was detected 1 day prior to ovulation. Irregular urinary progesterone metabolite profiles followed anovulatory cycles, reflecting varying degrees of follicular luteinization. In an attempt to ensure that a cycle would result in ovulation in the 5- to 6-year-old female, a GnRH treatment was tested during two separate cycles. Administration of GnRH on the day of behavioural oestrus resulted in an increase in urinary luteinizing hormone concentrations 2 h following injection. Regardless, ovulation did not occur in response to treatment. This study provides the first ultrasound data on ovarian activity in the Indian rhinoceros and establishes normal physiologic and behavioural relationships during the oestrous cycle that may facilitate the breeding of this species in captivity.

Rescue of the corpus luteum in human pregnancy.

Rescue of the corpus luteum from its programmed senescence maintains progesterone production required for pregnancy. In primates, chorionic gonadotropin produced by the developing conceptus acts as the primary luteotrophic signal. The purpose of this research was to assess corpus luteum rescue by examining changes in daily urinary progesterone metabolite levels during the first week after implantation. We determined the variability in progesterone metabolite profiles and evaluated its relationship to early pregnancy loss in 120 naturally conceived human pregnancies, including 43 early pregnancy losses. In other primates, an abrupt increase in the progesterone metabolite occurs at the time of implantation. This pattern occurred in an estimated 45% of the pregnancies in the present study. In the remaining pregnancies, there was a delayed rise (18%), neither a rise or decline (22%), or a decline (15%) during the week after implantation. The estimated rate of early pregnancy loss increased across these categories (from 5% loss with an abrupt rise at implantation to 100% loss with progesterone metabolite decline). Low urinary hCG levels in early pregnancy were significant determinants of a decline in postimplantation progesterone metabolite. However, preimplantation steroid metabolite levels were not significant, suggesting no inherent problem with the corpus luteum. Examination of individual progesterone metabolite profiles in relation to hCG profiles also indicated that few losses were caused by corpus luteum failure. Delineating the functional importance of an abrupt progesterone rise at the time of implantation may provide new strategies for promoting successful implantation in assisted reproduction.

Non-invasive monitoring of ovarian function in Asian elephants (Elephas maximus) by measurement of urinary 5 -pregnanetriol

The development of an enzymeimmunoassay for 5 beta-pregnanetriol and its use for non-invasive monitoring of reproductive cycles in Asian elephants is described. Gas chromatography-mass spectrometry (GCMS) and high performance liquid chromatography (HPLC) confirmed the presence of 5 beta-pregnane-3 alpha,17 alpha,20 alpha/beta-triols as the two most abundant urinary progesterone metabolites. The assay developed used the antiserum anti-5 beta-pregnane-17 alpha,20 alpha-diol-3 alpha-gamma l glucuronide but was designed to measure the free steroid in urine samples after hydrolysis and extraction. HPLC confirmed the presence of immunoreactive pregnanetriol in urine, but indicated that the measurement was nonspecific. Immunoreactive pregnanetriol concentrations were significantly correlated with the concentrations of both progesterone (r = 0.98, n = 269, P < 0.01) and 17 alpha-hydroxyprogesterone (r = 0.95, n = 205, P < 0.01), the metabolic precursor of pregnanetriol. The mean +/- SEM deviation of cycles as determined by measurements of plasma progesterone, 17 alpha-hydroxyprogesterone and urinary pregnanetriol, respectively, were 15.54 +/- 1.5 (n = 23, where n = number of cycles), 15.21 +/- 1.7 (n = 15) and 15.45 +/- 0.94 weeks (n = 20). These results demonstrate that it is possible to monitor ovarian function in Asian elephants by the measurement of urinary immunoreactive pregnanetriol concentrations.

Oestrous cycle of the North American bison (Bison bison) characterized by urinary pregnanediol-3-glucuronide

An enzyme immunoassay for urinary pregnanediol-3 alpha-glucuronide (PdG) was evaluated for the indirect measurement of progesterone metabolites during the oestrous cycle and early pregnancy of uncaptured North American bison. Comparisons between plasma progesterone and urinary PdG, dose-response parallelism between the standard curve and diluted urine samples and high-performance liquid cochromatography revealed that PdG was a primary immunoreactive urinary metabolite of progesterone in bison. Urine samples were collected directly from the soil from 29 bison cows during the August rutting season and analysed for PdG. Eight bison cows demonstrated complete oestrous cycles ranging from 19 to 26 days (mean cycle length = 23.12 +/- 0.76 days) and behavioural oestrus among four of these cows correlated with PdG nadirs. Mean PdG nadirs were 63.62 +/- 21.61 ng/mg urinary creatinine (Cr) and mean peak midluteal values were 546.01 +/- 130.73 ng/mg Cr. Seven of eight became pregnant, indicating that bison exhibit a second seasonal oestrus. Eighteen other bison cows were pregnant prior to the beginning of the study and demonstrated non-cyclic increased PdG concentrations (greater than 200 ng/mg Cr) during the 30-day course of collection. Three cows ovulated and became pregnant during the 30-day collection period and then exhibited increasing urinary PdG concentrations. This report demonstrates that ovarian function in uncaptured bison can be monitored by means of urinary PdG and that both ovulatory cycles and early pregnancy can be detected.

Urinary steroid evaluations to monitor ovarian function in exotic ungulates: VII. Urinary progesterone metabolites in the Equidae assessed by immunoassay

AbstractA direct enzyme immunoassay (EIA) for non‐specific urinary progesterone (Po) metabolites, utilizing a non‐specific monoclonal antibody against pregnanediol‐3‐glucuronide, was evaluated for the purpose of assessing luteal function in equids. Urinary pregnanediol‐3‐glucuronide (PdG) and immunoreactive PdG‐like conjugate (iPdG) concentrations, indexed by creatinine, were compared to plasma Po concentrations in non‐conceptive ovarian cycles through two ovulations in four mares. High‐performance liquid chromatography (HPLC) of urine from lutealphase mares and a pregnant zebra revealed an absence of significant concentrations of PdG and the presence of at least three immunoreactive compounds, all of which were more polar than PdG. The concentration of iPdG in the mare ranged from a nadir of approximately 3 ng/mg Cr at the time of ovulation to nearly 400 ng/mg Cr at the mid‐luteal‐phase peak and paralleled plasma Po concentrations. This non‐radiometric assay for iPdG permits the assessment of ovulation, luteal formation and function, and luteolysis in unprocessed urine samples from domestic mares. Data from a single zebra indicate this approach also will permit simplified and non‐invasive longitudinal studies of ovarian function among a wide range of Equidae.

Progesterone, some progesterone derivatives and urinary digoxin-like substances from pregnant women in radioimmuno- and 86Rb-uptake assays of digoxin.

Progesterone and some derivatives were tested in a radioimmunoassay (RIA) of digoxin and in a bioassay measuring the 86Rb-uptake into red blood cells as an index of Na+, K+-ATPase activity. The digitalis-like activity of the hormones was compared with that found in chromatographic fractions of material extracted from the urines of pregnant women at term. Progesterone at concentrations greater than 10(-6) M cross-reacted in the RIA, and at 10(-3) M it decreased 86Rb-uptake by 18%. The anaesthetic progesterone derivates 5 alpha-pregnane-3 alpha-ol-20-one and 5 alpha-pregnane-3,20-dione crossreacted to a lesser degree in the RIA and lacked effect in the bioassay. Similar results were obtained with pregnandiol-glucuronide, the major urinary metabolite of progesterone. In contrast, several fractions of the urinary material had significant effects in both assays. It is concluded that the digitalis-like activity of progesterone is not coupled to properties associated with its anaesthetic effects. Furthermore, although progesterone may account for a part of the endogenous digoxin-like substances in serum of neonates and pregnant women, neither progesterone proper nor pregnandiol-glucuronide explains the great amount of digoxin-like substances found in the urines.

Metabolic profiles of endogenous and ethynyl steroids in plasma and urine from women during administration of oral contraceptives

Conjugated ethynyl and endogenous steroids in plasma and urine from two women taking an oral contraceptive (Conlumin ®) containing 1 mg norethindrone and 50 μg mestranol have been analyzed by methods based on aruon and ligand exchange chromatography and gas chromatography-mass spectrometry. Conjugated norethindrone and its reduced metabolites with 3α,5α, 3α,5β, 3β,5β and 3β,5α configurations were identified in the fluids. The quantitatively major metabolites in plasma were a disulphate of the 3α,5α isomer and a monosulphate of the 3α,5β isomer. The renal clearance of the former compound was low. The major urinary metabolite of norethindrone was the 3α,5β isomer conjugated with glucuronic or sulphuric acid. Disulphates constituted only a small portion of urinary ethynyl steroids. Metabolic profiles of endogenous neutral steroids in plasma and urine during the contraceptive cycle were compared with profiles during a physiological menstrual cycle. The concentrations of steroids in plasma during contraception were similar to those during the follicular and mid phases of the menstrual cycle, whereas levels of progesterone metabolites were higher in the luteal phase. The urinary excretion of steroids was 15–30% lower during the contraceptive cycle, due to a decrease in excretion of C 21O 5 steroids, 11-oxygenated androgens and etiocholanolone. The increase of urinary progesterone metabolites seen during the luteal phase was not observed during contraception, but the excretion of 5β-pregnane-3α,20α-diol glucuronide was higher than during the follicular and mid phases of the menstrual cycle.

The detection of ovulation and early pregnancy in the baboon by direct measurement of conjugated steroids in urine.

The pattern of excretion of urinary steroid metabolites in the olive baboon (Papio anubis) was examined during the menstrual cycle and in conception cycles in which embryos were surgically removed at intervals between day 11 and day 21 (day 0 = day of preovulatory estrogen peak). Conjugated estrone and pregnanediol-3α-glucuronide were measured in overnight urine samples by direct, nonextraction assays, and the levels were indexed by creatinine. Results showed that measurement of urinary conjugated estrone reflected preovulatory estrogen output and that pregnanediol-3α-glucuronide was an abundant urinary metabolite of progesterone. There was a defined postovulatory increase in the excretion of conjugated estrone during conception cycles in eight of ten animals. The timing of the increase ranged between day 13 and day 19 and was related to the appearance of elevated levels of urinary gonadotrophin. In four animals, increased estrogen excretion was first detected after the day of embryo removal, but this was most likely a response to chorionic gonadotrophin secreted before surgery. The findings demonstrate that measurement of conjugated estrone offers a rapid and practical approach for monitoring ovulation and implantation in the baboon by a single assay technique.

Monitoring luteal function in the lion‐tailed macaque (Macaca silenus) through urinary progesterone metabolite measurements

AbstractA direct radioimmunoassay for measuring urinary 20‐hydroxyprogesterone cross‐reactivity to monitor and assess luteal function and detect pregnancy in the lion‐tailed macaque (Macaca silenus) is described. Urine samples were collected daily during ten nonconceptive and five conceptive ovarian cycles of five dult female lion‐tailed macaques. Urine was analyzed for concentrations of 20α‐hydroxypro‐gesterone cross‐reactivity, estrone conjugates, and creatinine.The strength of the luteal phase in normal nonconceptive cycles (n = 8) is characterized by a maximum sevenfold increase (day 9) in mean 20α‐hydroxyprogesterone cross‐reactivity over follicular phase levels; the duration, by a 13‐day sustained elevation of mean 20α‐hydroxyprogesterone cross‐reactivity levels. Pregnancy is detectable from 20α‐hydroxyprogesterone cross‐reactivity values approximately 20 days after the periovulatory estrone conjugate peak (n = 4). Apparent anovulation (n = 1), extended follicular phase (n = 1), and early abortion (n = 1) also are detectable using 20α‐hydroxyprogesterone cross‐reactivity measurements.

Aspects of the reproductive endocrinology of the female Giant panda (Ailuropoda metanoleaca) in captivity with special reference to the detection of ovulation and pregnancy

The metabolism and pattern of excretion of urinary steroids during oestrus and pregnancy in the Giant panda is described. Three female pandas from the London, Washington and Madrid Zoos were studied over different periods between March 1980 and September 1982.High pressure liquid chromatography and sequential enzyme hydrolysis indicated that oestrone glucuronide was the most abundant urinary oestrogen metabolite during oestrus. Levels of conjugated oestrone in late pregnancy, however, were low and similar to those of conjugated oestradiol‐17βThere was a rapid increase in the excretion of conjugated oestrone to reach maximum levels during late pro‐oestrus; oestrus occurred when levels of conjugated oestrone were declining. The measurement of oestrone‐3‐glucuronide by direct, non‐extraction assay provides a rapid and reliable method for detecting oestrus and ovulation in the Giant panda.Artificial insemination of the London and Madrid pandas was performed in 1981 and 1982, respectively. The Madrid panda gave birth to twin cubs after a gestation period of 159 days. Levels of urinary oestrone conjugate remained low throughout pregnancy. There was no increase in the excretion of pregnanediol‐3α‐glucuronide (assumed to be an urinary metabolite of progesterone) until approximately day 120 when a rapid, five‐fold increase in levels occurred. The levels of pregnanediol‐3α‐glucuronide remained elevated for approximately three weeks after which there was a gradual decline beginning two‐and‐α‐half weeks before parturition. Measurement of pregnanediol‐3α‐glucuronide enables the detection of pregnancy after three to four months and should also be useful in predicting parturition. A delay of implantation during pregnancy in the Giant panda is suggested.There was no consistent elevation in pregnanedioI‐3α‐glucuronide excretion in the five months after artificial insemination of the London panda, despite a marked increase in circulating progesterone of ovarian origin. Pregnancy could not be confirmed from external examination of the uterus at laparotomy; histological examination of biopsy material revealed advanced endometrial hyperplasia.