Pregnanediol Glucuronide Levels Research Articles

Urinary reproductive hormone level differences between African American and Caucasian women of reproductive age

Objective: To compare urinary levels of reproductive hormones in African American and Caucasian women. Design: Cross-sectional study. Setting: Ten United States Air Force (USAF) bases. Patient(s): African American (n = 33) and Caucasian (n = 65) women of reproductive age from a larger study of USAF women (n = 170). Intervention(s): None. Main Outcome Measure(s): Urinary endocrine end points: follicular luteinizing hormone (LH), preovulatory LH, level of LH surge peak, early follicular follicle stimulating hormone (FSH), follicular LH:FSH ratio, midluteal FSH, FSH rise before menses, early follicular estrone 3-glucuronide (E 13G), midfollicular E 13G, periovulatory E 13G peak, midluteal E 13G, early follicular pregnanediol 3-glucuronide (Pd3G), follicular Pd3G, rate of periovulatory Pd3G increase, E 13G:Pd3G on the day of luteal transition, slope of E 13G:Pd3G, and midluteal Pd3G. Result(s): Relative to Caucasians, African American women had significantly lower follicular phase LH:FSH ratios (mean ± SD: 0.7 ± 0.4 vs. 1.0 ± 0.6), lower follicular phase Pd3G levels (1.0 ± 0.5 vs. 1.2 ± 0.8 μg/mg creatinine), and lower rates of periovulatory Pd3G increase (0.5 ± 0.7 vs. 1.0 ± 1.2 μg/mg creatinine). Conclusion(s): Findings of this analysis should be considered preliminary evidence of racial differences in hormone levels. Future studies are needed to determine whether these differences have clinical significance.

Mean versus individual hormonal profiles in the menstrual cycle

Objective: To evaluate hormonal profiles of normal menstrual cycles. Design: Prospective, descriptive study of a case series. Setting: University-based natural family planning center. Patient(s): Twenty-five natural family planning users for three or more cycles (n = 78). These women were healthy, contraception-free, parous, with regular ovulatory cycles. Intervention(s): Immunoassays for estrone glucuronide, LH, and pregnanediol glucuronide were done in daily timed and measured samples of early morning urine. Main Outcome Measure(s): Estrone glucuronide, LH, and pregnanediol glucuronide levels were measured during the menstrual cycle. Result(s): All cycles showed an ovulatory pattern configuring classic hormonal mean curves. Most (77%) differed from the mean curve pattern. All had estrone glucuronide peaks, LH peaks, and pregnanediol glucuronide increases. Estrone glucuronide and LH peaks were not always clear; some lasted more than 1 day (long peak: estrone glucuronide 19%, LH 9%) or fluctuated (double peak: estrone glucuronide 4%, LH 6%; small LH peak: 19%). There were also prepeak estrone glucuronide surges, and pre- and postpeak LH surges. Pregnanediol glucuronide increased more clearly (6% fluctuated 1 day). Some women had repeated cycles with long estrone glucuronide peaks (16%) and fluctuations in LH surge (44%). Conclusion(s): Normal menstrual cycle hormonal profiles generally differ from mean curves, which are usually considered standard.

Feasibility of determination of fertile period in Indian women based on urinary hormone estimations.

Baseline data available on the excretory profiles of estrone giucuronide (E(1)G), pregnanediol glucuronide (PdG) and luteinising hormone (LH) on human menstrual cycles (n=104) was retrospectively analysed for identifying the limits of fertile period (FP) to be used as natural method of family planning. The limits of fertile period are suggested based on centile distribution of E(1)G and PdG levels during defined phase of menstrual cycle. Two approaches, which do not involve any mathematical calculation are suggested. In approach A, fertile period is said to have started when E(1)G value of 35 ng/ml is reached and is said to have ended when the PdG value of 2 μg/ml on two consecutive days is obtained. The criteria were applied to 30 test cycles in whom authentic fertile period was identified based on excretory profiles of E(1)G, PdG and LH throughout the menstrual cycles. When approach A was followed the authentic fertile period was covered in 27 cycles giving an accuracy of 90% with a mean fertile period length of 9.11+1.9 days. In approach B, the cut off limit of E(1)G value was increased to 55 ng/ml in order to reduce the days of abstinence. Though the length of the fertile period was reduced to 7.2+1.5 days the accuracy of the approach was 66.6%. Thus the approach A which has accuracy of 90% may appeal to determined couples who wish to practice family planning by periodic abstinence or restrict the use of barrier methods.

The effect of small doses of progesterone released from two types of vaginal rings on ovarian activity and bleeding patterns during the first postpartum year

A total of 20 breastfeeding women used progesterone-releasing vaginal rings for up to 12 months. The women were divided into two groups, one (n = 9) using rings with an initial release rate of 5 mg/24 h, the other (n = 11) with a release rate of 20 mg/24 h. Individual women started to give breast milk supplements and ceased to breastfeed after various periods of time. Urinary estrone and pregnanediol glucuronide levels were measured by radioimmunoassay three times weekly during the entire trial. Individual diary cards were used to register bleeding and spotting. A significantly higher concentration of pregnanediol glucuronide was seen when the 20 mg/24 h ring was used in the lactation period, in comparison with the 5 mg/24 h ring. In the post-lactation period, pregnanediol glucuronide levels dropped when the 20 mg/24 h ring was used. Estrone glucuronide levels increased after the termination of breastfeeding, indicating an enhanced suppression of ovarian activity in the lactation period with both rings. Although the degree of suppression was dose-related, both rings were likely to offer a sufficient contraceptive effect in the lactation period. No significant changes were observed when milk supplements were added to breastfeeding. The use of the 20 mg/24 h ring resulted in a much better bleeding pattern (significantly less bleeding days) than the 5 mg/24 h ring during the lactation period. In the post-lactation period, the 20 mg/24 h ring the bleeding became much worse than in the lactation period. As a result of this investigation, use of the 20 mg/24 h ring during the lactation period could be recommended and, if unacceptable bleeding occurs after the termination of breastfeeding, the ring can be replaced by another contraceptive method.

Endocrine monitoring of the ovarian cycle and pregnancy in the saddle-back tamarin (Saguinus fuscicollis) by measurement of steroid conjugates in urine.

Direct measurements of urinary immunoreactive estrone conjugates (E1C) and pregnanediol glucuronide (PdG), were applied to monitoring the ovarian cycle (n = 9) and pregnancy (3 full term pregnancies, 2 mid-term abortions) in Saguinus fuscicollis. During the ovarian cycle, urinary E1C concentrations revealed a high degree of day-to-day variability and appeared to be uninformative in reflecting cyclic ovarian function. In contrast, PdG was a reliable indicator of ovarian cyclicity with excretion patterns corresponding well with plasma progesterone profiles. Luteal phase PdG concentrations were on average 4-7-fold higher than corresponding follicular phase values. On the basis of changes in circulating progesterone, a mean cycle length of 25.7 ±1.0 days with an average follicular phase of 7.1 ± 0.6 days and a mean luteal phase of 18.6 ± 0.7 days, was found (n = 14 cycles). Following conception, both urinary steroid conjugate concentrations increased and elevated levels were maintained beyond the normal luteal phase length, allowing pregnancy to be determined at around day 25-30. During mid- to late pregnancy, PdG levels declined while E1C concentrations continued to be elevated until approximately 6 weeks before parturition when a decrease occurred. Both hormones showed a clear and rapid fall to follicular phase values following termination of pregnancy at either parturition or mid-term abortion. Post partum ovulations (n = 5) occurred on average 17-18 days following birth with four ovulations leading to conceptions. The results demonstrate the potential of urinary steroid conjugate analysis as a practical and reliable method for non-invasive monitoring of reproductive status in the female saddle-back tamarin. © 1995 Wiley-Liss, Inc.

Excretory profile of inhibin-like peptide (ILP) during human menstrual cycle: a probable marker for determination of fertile period

The excretory profile of inhibin-like peptide (10.4 kDa) and its interrelationship with urinary LH, FSH, oestrone glucuronide (E 1G) and pregnanediol glucuronide (PdG) during the menstrual cycle were studied. These hormones/metabolites were estimated in daily early morning urine samples obtained from 20 regularly menstruating women. The data revealed that the excretory profile of inhibin-like peptide (ILP) follows a pattern similar to that of E 1G. In 17 cycles, ILP peaked 3–4 days prior to the urinary LH peak. A value of 70 ng/mg creatinine (95th centile of ILP levels obtained between 2 and 4 days prior to urinary LH peak and also 5th centile of peak ILP levels) was considered as an indicator of the start of the fertile period. A value of PdG more than 2 μg/mg creatinine on two consecutive days was considered as an end of the fertile period. The entire fertile period could be determined in 18 out of 20 cycles when criteria based on ILP and PdG levels were applied (accuracy, 90%), whereas it could be determined in 13 out of 20 cycles when criteria based on E 1G and PdG levels were applied (accuracy, 65%). Thus, ILP levels in urine may prove to be one of the signals for determining the start of fertile period.

First-trimester rapid semiquantitative assay for urine pregnanediol glucuronide predicts gestational outcome with the same diagnostic accuracy as serial human chorionic gonadotropin measurements

OBJECTIVE: The purpose of this study was to compare a single urine pregnanediol glucuronide measurement with serial human chorionic gonadotropin titers for the prediction of abnormal early gestations. STUDY DESIGN: We analyzed multiple urine pregnanediol glucuronide levels in 19 spontaneously conceived pregnancies during the first 49 days of gestation. A semiquantitative measurement was made by rapid enzyme immunoassay (Phase Check) at different urinary dilutions. To establish the reliability of semiquantitative urine pregnanediol glucuronide assay to detect abnormal gestation, this test was compared with human chorionic gonadotropin doubling times derived from a previously described normal population. A receiver-operator characteristic curve was constructed for each test, and areas under the curve with corresponding SEs were calculated. The critical-ratio z test was used to compare the two assays. RESULTS: The receiver-operator characteristic curves indicate that both urine pregnanediol glucuronide and human chorionic gonadotropin doubling can predict early gestational complications (p < 0.05). The area under the curve for human chorionic gonadotropin doubling time was 0.809 ± 0.048, and urine pregnanediol glucuronide had an area of 0.702 ± 0.072. Comparison of the area under the curve revealed that the ability of urine pregnanediol glucuronide to predict early gestational failure was indistinguishable from that of human chorionic gonadotropin doubling times (p > 0.05). A 1:2 dilution of urine gave the best results in the semiquantitative urine pregnanediol glucuronide test (Phase Check). CONCLUSIONS: On the basis of receiver-operator analysis, semiquantitative urine pregnanediol glucuronide measurements predict abnormal early gestations as well as serial human chorionic gonadotropin measurements do. The ability of a single urine semiquantitative assay (Phase Check) to predict early gestational complications offers a convenient screening tool that may identify women with abortive or ectopic pregnancies before the onset of symptoms. (AM J Obstet Gynecol 1994;170:1822-7.)

First-trimester rapid semiquantitative assay for urine pregnanediol glucuronide predicts gestational outcome with the same diagnostic accuracy as serial human chorionic gonadotropin measurements

Objective: The purpose of this study was to compare a single urine pregnanediol glucuronide measurement with serial human chorionic gonadotropin titers for the prediction of abnormal early gestations. Study Design: We analyzed multiple urine pregnanediol glucuronide levels in 19 spontaneously conceived pregnancies during the first 49 days of gestation. A semiquantitative measurement was made by rapid enzyme immunoassay (Phase Check) at different urinary dilutions. To establish the reliability of semiquantitative urine pregnanediol glucuronide assay to detect abnormal gestation, this test was compared with human chorionic gonadotropin doubling times derived from a previously described normal population. A receiver-operator characteristic curve was constructed for each test, and areas under the curve with corresponding SEs were calculated. The critical-ratio z test was used to compare the two assays. Results: The receiver-operator characteristic curves indicate that both urine pregnanediol glucuronide and human chorionic gonadotropin doubling can predict early gestational complications (p < 0.05). The area under the curve for human chorionic gonadotropin doubling time was 0.809 ± 0.048, and urine pregnanediol glucuronide had an area of 0.702 ± 0.072. Comparison of the area under the curve revealed that the ability of urine pregnanediol glucuronide to predict early gestational failure was indistinguishable from that of human chorionic gonadotropin doubling times (p>0.05). A 1:2 dilution of urine gave the best results in the semiquantitative urine pregnanediol glucuronide test (Phase Check). Conclusions: On the basis of receiver-operator analysis, semiquantitative urine pregnanediol glucuronide measurements predict abnormal early gestations as well as serial human chorionic gonadotropin measurements do. The ability of a single urine semiquantitative assay (Phase Check) to predict early gestational complications offers a convenient screening tool that may identify women with abortive or ectopic pregnancies before the onset of symptoms. Objective: The purpose of this study was to compare a single urine pregnanediol glucuronide measurement with serial human chorionic gonadotropin titers for the prediction of abnormal early gestations. Study Design: We analyzed multiple urine pregnanediol glucuronide levels in 19 spontaneously conceived pregnancies during the first 49 days of gestation. A semiquantitative measurement was made by rapid enzyme immunoassay (Phase Check) at different urinary dilutions. To establish the reliability of semiquantitative urine pregnanediol glucuronide assay to detect abnormal gestation, this test was compared with human chorionic gonadotropin doubling times derived from a previously described normal population. A receiver-operator characteristic curve was constructed for each test, and areas under the curve with corresponding SEs were calculated. The critical-ratio z test was used to compare the two assays. Results: The receiver-operator characteristic curves indicate that both urine pregnanediol glucuronide and human chorionic gonadotropin doubling can predict early gestational complications (p < 0.05). The area under the curve for human chorionic gonadotropin doubling time was 0.809 ± 0.048, and urine pregnanediol glucuronide had an area of 0.702 ± 0.072. Comparison of the area under the curve revealed that the ability of urine pregnanediol glucuronide to predict early gestational failure was indistinguishable from that of human chorionic gonadotropin doubling times (p>0.05). A 1:2 dilution of urine gave the best results in the semiquantitative urine pregnanediol glucuronide test (Phase Check). Conclusions: On the basis of receiver-operator analysis, semiquantitative urine pregnanediol glucuronide measurements predict abnormal early gestations as well as serial human chorionic gonadotropin measurements do. The ability of a single urine semiquantitative assay (Phase Check) to predict early gestational complications offers a convenient screening tool that may identify women with abortive or ectopic pregnancies before the onset of symptoms.

Hypothalamic-pituitary-ovarian response to clomiphene citrate in women with polycystic ovary syndrome

To examine the hypothalamic-pituitary sites of clomiphene citrate (CC) action in women with polycystic ovarian syndrome (PCOS). Prospective controlled trial. Seventeen women with PCOS and 9 normal-cycling women. Subjects with PCOS received CC, 150 mg/d for 5 days. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels and LH pulse characteristics and their response to gonadotropin-releasing hormone (GnRH, 10 micrograms) were examined before and after 3 days of CC in PCOS subjects during a 12-hour frequent sampling study (n = 8). Daily urinary estrone glucuronide and pregnanediol glucuronide levels after CC were compared with concentrations in normal-cycling women through one menstrual cycle. In another nine PCOS subjects, pituitary and ovarian hormonal cyclicity was monitored by daily blood sampling. Thirteen of 17 treated cycles were ovulatory with normal luteal phases. In the ovulatory cycles, serum LH, FSH, estradiol (E2), and estrone levels increased after CC. Luteinizing hormone pulse frequency was unchanged, but LH pulse amplitude increased significantly after CC. Both LH and FSH response to exogenous GnRH was significantly attenuated after CC treatment. In anovulatory cycles, serum LH, FSH, and E2 increased initially and then returned to baseline and remained unchanged for the ensuring 40 days. Clomiphene citrate-induced ovulation in women with PCOS is accompanied by increased secretion of LH and FSH with enhanced estrogen secretion. The increased LH pulse amplitude after CC, together with decreased pituitary sensitivity to GnRH, suggests a hypothalamic effect.

Inhibition of ovulation by low-dose mifepristone (RU 486).

Mifepristone (RU 486) is a potent antigestagen and antiglucocorticoid which when given at a dose of 25-600 mg disrupts folliculogenesis, inhibits ovulation and induces menses in healthy women. This study reports the effects of much lower doses of mifepristone than used previously, given for the duration of a complete menstrual cycle. Healthy female volunteers (n = 11) with regular menstrual cycles were given mifepristone at a daily dose of 5 mg (n = 6) or 2 mg (n = 5) for 30 days, beginning immediately after an ovulatory placebo cycle. Mifepristone prevented menstruation for the duration of the treatment period, with recurrence of menses 15-29 days after replacement of mifepristone with placebo. Daily mifepristone given in either 5 mg or 2 mg doses inhibited ovulation, as indicated by the lack of a rise in urinary pregnanediol excretion. The excretion of oestrone glucuronide in urine rose during treatment, suggesting ovarian follicular development. Inhibition of ovulation appeared to result from a failure of the positive feedback effect of oestradiol on the hypothalamo-pituitary axis, as no surges of luteinizing hormone were seen despite pre-ovulatory levels of oestrone glucuronide being measured during exposure to mifepristone. The cycle immediately following treatment was shorter than the pre-treatment cycle, with lower peak levels of pregnanediol glucuronide, suggesting an inadequate luteal phase. Recovery from the effects of mifepristone treatment was more rapid after 2 mg than after 5 mg and one subject conceived in the immediate post-treatment phase, indicating adequate ovulation and luteinization.(ABSTRACT TRUNCATED AT 250 WORDS)

Monitoring the ovarian cycles of Pan troglodytes and P. paniscus: A comparative approach.

Females of both species of chimpanzee, Pan troglodytes and P. paniscus, exhibit conspicuous anogenital sex swellings that are useful external markers for assessing ovarian cyclicity. A new, quantitative method for describing these swelling cycles was refined during laboratory studies of four P. paniscus and nine P. troglodytes and was used to analyze variability in swelling patterns. Clear-cut anatomical changes of the perineum were found to have both interindividual and interspecific reliability. The swollen phase of the cycle was defined as the period when the medial surfaces of the labia minora are pressed against each other so the aperture leading to the vagina appears as a slit (labial occlusion). When defined with this precision, comparisons within and between the species revealed two types of cycle for both species; swollen phases of P. paniscus were about 24 or 39 days in duration, on average, whereas those of P. troglodytes were about 14 or 18 days. Urine samples from the P. troglodytes were collected and analyzed for luteinizing hormone (LH) (34 cycles), estrone glucuronide (E1G), and pregnanediol-3-glucuronide (PdG) (18 cycles). When the two types of cycle were separated in analysis, it was found that the day when labial occlusion occurred served as a predictor of the preovulatory LH peak in the cycles with a long swollen phase. Levels of E1G correlated closely with swelling scores in multiparous females regardless of phase lengths, suggesting that the new swelling scores are a reliable indicator of E1G excretion in multipara. Cycles with long swollen phases had relatively high levels of PdG during the luteal phase, whereas cycles with short swollen phases had relatively low levels of PdG and a shorter luteal phase. The marker of labial occlusion and the duration of the preswollen phase defined by the marker appear to be potentially useful in studies of reproduction in Pan, for monitoring female fertility, and in artificial insemination procedures. If it is confirmed that swelling pattern predicts progesterone levels during the luteal phase, it would make interventions possible in the case of low levels so that a potential pregnancy could be maintained to term.

Copulatory frequency, urinary pregnanediol, and fertility in great apes.

Previous research on sexual behavior of the great apes suggests that female chimpanzees readily conceive in traditional laboratory pair tests, i.e., free-access tests (FATS), but female gorillas and orang-utans do not. Although several different mechanisms could account for this result, the one tested is derived from R.V. Short's hypothesis that "form reflects function," with respect to the ratio of testis weight to body weight in the great apes. The implication of this hypothesis is that the more frequent maleinitiated mating that occurs prior to ovulation in the FATs, in comparison with the wild, reduces the fertility of male orang-utans and gorillas as a function of their relatively small testes. The hypothesis was tested by conducting restricted-access tests (RATS) with female choice, tests in which the female determines whether and when copulation may occur. In the RATs, the cycle rate of copulation was reduced in both the gorillas and orang-utans to rates typical of their wild counterparts. Three females of each species, moreover, did conceive in the RATs, but this result was not statistically significant. Several conditions were implicated in the failure of additional females of both species to conceive during the RATs, including an absence of copulation, copulation without ejaculation, and copulation with ejaculation temporally dissociated from the presumptive time of ovulation. Little support was obtained for the hypothesis that failure to conceive during the FATs was due to the males' relatively small testes and frequent copulation, but such a possibility cannot be ruled out. Relatively low levels of pregnanediol glucuronide during the luteal phase of the menstrual cycle were implicated in the failure to conceive by female gorillas but not female orang-utans. The data suggest that female regulation of mating in the RATs results in patterns and rates of copulation that are more characteristic of these species than those obtained in the FATs. Restricted-access testing thus may prove useful in facilitating the breeding of gorillas and orang-utans in other captive settings.

The development of an enzyme‐immunoassay for urinary pregnanediol‐3‐glucuronide and its application to reproductive assessment in exotic mammals

Measurement of urinary pregnanediol‐3‐glucuronide (PdG) provides a practical, non‐invasive method for monitoring ovarian function and pregnancy in diverse mammalian species. To date, however, methods of laboratory analysis have relied on radio‐immunoassay procedures which, because of their cost and the problems associated with the use and disposal of radioactive substances, have limited potential for routine application in zoos and other animal collections with limited laboratory facilities. The present paper reports the development of a non‐isotopic enzyme‐immunoassay for the measurement of urinary PdG in a wide range of exotic mammals and describes its application to reproductive assessment in the gorilla and rhinoceros.The assay is based on an homologous competitive binding system and utilizes antiserum raised against PdG‐bovine serum albumen and alkaline phosphatase‐PdG, prepared by the active ester technique, as conjugate. Sensitivity of the assay is between 12.5 and 25 pg/well and precision, expressed as the intra‐ and inter‐assay coefficients of variation, is between 5.3 and 7.8% and 9.2 and 16.1%, respectively. Serial dilutions of human, gorilla, orang‐utan, mangabey, blackbuck, okapi, gaur, giant panda and Indian, black and white rhinoceros urine gave displacement curves parallel to that of the PdG standard, indicating the potential of the assay for multi‐species application.The pattern of excretion of immunoreactive PdG during successive menstrual cycles and two complete pregnancies in a gorilla is described. Follicular phase levels of PdG were low (0.07–0.13 μg/mg creatinine (Cr)), in contrast to the high levels of excretion during the luteal phase (maximum values 1.5–2.1 μg/mg Cr). Mean cycle length was 29.6±1.5 days with a range of 26–34 (n = 5). The pattern of PdG excretion during pregnancy showed a slow increase during the first 60 days and a more rapid rise to values of 8–12 μg/mg Cr after 120 days, with no further consistent increase during the remaining period of pregnancy. Estimated gestation lengths were 280 and 256 days.The levels of immunoreactive PdG excreted during mid‐to‐late pregnancy varied considerably between the three species of rhinoceros, maximum values ranging from up to 10 μg/mg Cr in the Indian to 0.08 μg/mg Cr in the white rhinoceros. Despite this variation pregnancy was associated with elevated PdG excretion in all three species and each profile showed a marked fall in levels at the end of the gestation period. The results suggest potential for urinary PdG determination as a practical method of pregnancy detection in both Indian and African rhinoceros, although species differences in the relative importance of PdG as a progesterone metabolite probably exist.

The effect of tamoxifen on the function and lifespan of the corpus luteum and on subsequent ovarian function

The aim of the present study was to investigate the effects of tamoxifen on pituitary and luteal function and on the bleeding pattern when administered continuously in the secretory phase. The study included 16 women with regular menstrual cycles followed during one control, one treatment and one follow-up cycle. Each volunteer received 20 mg tamoxifen twice daily from cycle day 18 to menstruation in the treatment cycle. The luteal phase was slightly, but significantly prolonged during treatment, and FSH, progesterone, 17-hydroxyprogesterone, 20 alpha- dihydro progesterone, estrone, estrone sulphate and estradiol significantly elevated in comparison with corresponding data during the control cycle. The results indicate that estrogen may be of some importance for the regulation of the life span of the corpus luteum in the human. The significantly elevated levels of pregnanediol glucuronide and estrone glucuronide during the follow-up cycle are most likely a result of either a direct effect of remaining circulating tamoxifen levels on the ovary, or mediated through the increased release of FSH. If estrogens are of importance for the process of implantation, which has recently been suggested in sub-human primates, also in the human remains unclear. Studies on the effect of anti-estrogens on the endometrium during the secretory phase of the cycle are ongoing.

Urinary steroid concentrations during natural and gonadotrophin-induced oestrus and pregnancy in the giant panda (Ailuropoda melanoleuca).

Urinary concentrations of conjugated oestrone and pregnanediol-3-glucuronide were measured during and after spontaneous and induced oestrus and during pregnancy. Behavioural oestrus was preceded by a rise in oestrone values from less than 10 ng/mg creatinine (Cr) to peaks of 45 ng/mg Cr. Maximal lordotic response and mating activity coincided with the decline in oestrone levels. After presumed ovulation, urinary pregnanediol glucuronide concentrations increased from less than 5 to 15-30 ng/mg Cr. Further increases in this steroid (to 60-80 ng/mg Cr) occurred 114 days after mating, presumably coincident with implantation. These high levels of pregnanediol glucuronide were maintained for 3 weeks, began to decline 1 week before parturition and fell to a nadir (less than 5 ng/mg Cr) immediately after delivery. When FSH was administered i.m. for 5 days, urinary oestrone values rose markedly and were maximal (580 ng/mg Cr) on Day 7. Mating first occurred on Day 20 and 500 i.u. hCG were given i.m. Urinary pregnanediol glucuronide levels during the next 5 months were similar to those in the previous year during pregnancy with values rising 105-108 days after mating. However, no birth occurred. These results support the suggestion that pandas exhibit delayed implantation and demonstrate that the panda is responsive to exogenous gonadotrophins.

Urinary hormone analysis as a diagnostic tool to evaluate the ovarian function of female gorillas (Gorilla gorilla).

Daily urine samples were collected from 4 adult female gorillas over 7 menstrual cycles. Urinary oestrone conjugate and pregnanediol-3-glucuronide (PDG) were measured by radioimmunoassay; LH was measured by enzyme immunoassay and each hormone was indexed by creatinine. The quantity of urinary LH during the ovulatory surge was positively correlated with the quantity of PDG excreted during the luteal phase (r = 0.87, P = 0.0013). The observations indicate a relationship between the quality of the LH surge and the levels of PDG in the luteal phase and suggest that both the LH surge and subsequent luteal phase function may be predictable from the oestrogen excretion profile during the follicular phase.

Urinary steroid evaluations to monitor ovarian function in exotic ungulates: V. Estrogen and pregnanediol‐3‐glucuronide excretion in the black rhinoceros (Diceros bicornis)

AbstractA study of female black rhinoceros (Diceros bicomis) urinary steroid and steroid metabolite excretion was performed to determine if techniques useful for monitoring reproductive events in the Indian rhinoceros (Rhinoceros unicornis) could be utilized to evaluate the black rhinoceros. Urine samples from 19 zoo‐held black rhinoceros were analyzed for estrogen, estrone conjugates (EC), and pregnanediol‐3‐glucuronide (PDG) content by direct radioimmunoassays. Estrogen analysis revealed that &gt;95% of the estrogens present in female black rhinoceros urine are conjugated, with estrone and estradiol accounting for virtually all of these estrogens. There is no observable difference in the amount of estrogen present in estrus; postestrus; and early‐, mid‐, and late‐gestation urine samples. Analysis of serial urine samples for EC failed to reveal any discernible levels or patterns which reflected reproductive status. Neither nonpregnant nor early‐gestational female black rhinoceros' urine samples contained detectable amounts of PDG. Urinary PDG concentrations became measurable in midgestation (9–12 months prior to parturition) and rose steadily throughout the remainder of gestation. PDG levels declined sharply and became nondetectable 1 day postpartum. Though a wide range in PDG levels was observed among individual pregnant animals, each female consistently excreted increasing amounts of PDG through latter pregnancy.

Artificial insemination with frozen, thawed semen and pregnancy diagnosis in addax (Addax nasomaculatus)

AbstractNonsurgical artificial insemination (AI) procedures were performed using frozen, thawed semen in two adult female addax. Pregnant mares' serum gonadotropin (PMSG) (300 IU) and prostaglandin (PG) (125 μg) was administered intramuscularly to female 1 on days −3 and −1, respectively, where day 1 was the first day of AI. Only one injection of PG was used, as a low serum progesterone (PROG) concentration (0.5 ng/ml) and a low pregnanediol glucuronide (PdG) concentration (0.36 ng PdG/mg creatinine) had been recorded on day −13. A total of 219 × 106 motile sperm was deposited in the os cervix or anterior vagina on days 1–4. High serum PROG levels (9.4–15.8 ng/ml) and high urinary PdG levels (1.35–1.63 ng PdG/mg creatinine) were observed on days 23, 31, and 39, and the fetal heart was seen beating during an ultrasound exam on day 45. A male calf was born on day 266, 8.7 mo after insemination. This is the first live birth resulting from AI of an antelope with frozen, thawed semen. The Synchro‐mate‐B system of estrous control was used in female 2. The implant was inserted, and the injection was given on day −11 per the manufacturer's instructions. The implant was removed on day −1, and a total of 220 × 106 motile sperm was deposited in the os cervix or anterior vagina on days 1 and 2. High serum PROG levels (17.5–24.5 ng/ml) were observed on days 13, 21, and 29. Although the fetal heartbeat was observed during an ultrasound examinataion on day 42, the fetus was not carried to term. Details of the radioimmunoassay used to determine the luteinizing hormone immunoreactivity of urine samples are presented.

Urinary steroid evaluations to monitor ovarian function in exotic ungulates: II. Comparison between the giraffe (Giraffa camelopardalis) and the Okapi (Okapia johnstoni)

AbstractOvarian activity in the female giraffe was evaluated during the nonfertile ovarian cycle and during the terminal stages of gestation. Progesterone metabolites, in the form of pregnanediol‐3‐glucuronide (PdG), were measured in daily random urine samples collected from four adult parous giraffes. The follicular phase averaged 4.0 ± 0.1 days in length (N = 12; range 3–5 days) and peak PdG levels in the postovulatory period averaged 30.9 ± 1.7 ng/mg Cr (N = 12). PdG levels during the latter half of gestation greatly exceeded average luteal phase levels, which is in contrast to domestic ruminants. Prior to parturition, a marked decline in PdG excretion was evident, which may be useful for anticipating this event. These data serve to elucidate ovarian function in the mature female giraffe and to provide information concerning the physiologic role of certain anomalous ovarian structures. In addition, observed similarities in the pattern of PdG excretion during the nonfertile cycles of the giraffe and the okapi indicate similar underlying physiologic processes.

Urinary steroid evaluations to monitor ovarian function in exotic ungulates: III. Estrone sulfate and pregnanediol‐3‐glucuronide excretion in the Indian rhinoceros (Rhinoceros unicornis)

AbstractA study was undertaken to identify urinary estrogen and progesterone metabolites in the female Indian rhinoceros (Rhinoceros unicornis). Measurements of these metabolites were then used to monitor ovarian function and establish normal levels and patterns of steroid excretion during the estrous cycle and pregnancy. Urine samples were analyzed for estrone sulfate and pregnanediol‐3‐glucuronide (PDG) by direct radioimmunoassays. Both hormones produced discrete profiles reflecting ovarian activity in nonconceptive cycles. The estrous cycle was observed to be 48 days (range 39–64) with a mean follicular phase of 14.8 days (range 13–19), followed by a mean luteal phase of 19 days (range 17–21). Of the single gestation monitored, PDG levels rose above luteal phase levels by the third month after breeding and remained elevated throughout gestation. The combined estrogen and progesterone metabolite profiles present a complete evaluation of ovarian steriod production in the mature female Indian rhinoceros.