Urinary Hormone Analysis Research Articles

Direct-to-consumer fertility testing: utilization and perceived utility among fertility patients and reproductive endocrinologists

Research QuestionWhat is the utilization of direct-to-consumer fertility tests (DTCFT) among fertility patients? How does the perceived utility of DTCFT differ between patients and reproductive endocrinologists (REI)? DesignInfertility patients visiting the Duke Fertility Center between December 2020 and December 2021 were sent an electronic invitation to participate in a patient survey. Members of the Society of Reproductive Endocrinology and Infertility were also sent e-mail invitations to participate in the REI survey. DTCFT were defined as tests not ordered by a physician or performed at a physician's office, including calendar methods of ovulation prediction, urinary ovulation prediction kits, basal body temperature (BBT) monitoring, hormone analysis, ovarian reserve testing and semen analysis. Patients and REI were asked how likely they were to recommend a given DTCFT, on a 0–10 Likert scale. ResultsIn total, 425 patients (response rate 50.5%) and 178 REI (response rate 21.4%) completed the surveys. Patients reported the utilization of calendar methods of ovulation prediction (83.8%), urinary ovulation prediction (78.8%), BBT monitoring (30.8%), hormone analysis (15.3%), semen analysis (10.1%) and ovarian reserve testing (9.2%). REI rated the utility of all DTCFT significantly lower than patients did (average discordance –4.2, P < 0.001), except for urinary ovulation prediction, which REI gave a significantly higher score (discordance +1.0, P < 0.001). Prior pregnancy was significantly associated with home ovulation prediction utilization among patients (adjusted odds ratio 3.21, 95% confidence interval 1.2–9.83). ConclusionsMethods of ovulation prediction are commonly used by fertility patients. Significant discordance exists in the perceived utility of DTCFT between patients and REI. Patient education and guidelines are needed to better inform individuals considering DTCFT.

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.

Use of urinary biomarkers of ovarian function and altrenogest supplementation to enhance captive breeding success in the Indian rhinoceros (Rhinoceros unicornis)

Urinary hormone analysis was conducted on two adult female Indian rhinoceroses (Rhinoceros unicornis) that exhibited minimal or no estrual behaviors traditionally used to time breeding. Urine was collected throughout two consecutive estrous cycles to establish preliminary data on each individual's pattern and concentration of estrogen conjugates (EC) and progesterone metabolites (PdG) during follicular and luteal phases. Following preliminary endocrine analysis, urine samples were shipped on a frequent basis to verify when each female was off baseline in EC. Estrus and breeding dates were then predicted. Females were introduced to fresh male rhinoceros fecal samples daily throughout the follicular phase to potentially stimulate estrous behaviors. Despite successful assessment of follicular phase dynamics, females sometimes failed to exhibit estrus. Both females conceived following mating introductions that were timed using hormone analysis. Pregnancy was diagnosed either by endocrine analysis or rectal ultrasonography. Progestational support (altrenogest) occurred after pregnancy confirmation and varied for each female (21 and 66 days post-breeding). One female experienced early pregnancy loss and the other successfully completed a term pregnancy. These results demonstrate that a science based management strategy that relies on urinary biomarkers of ovarian function can facilitate naturally breeding captive Indian rhinoceroses.

Urinary hormone metabolites identify sex and imply unexpected winter breeding in an endangered, subterranean-nesting frog

Urinary hormone analysis has proved accurate for identifying sex and breeding periods in dimorphic amphibians with known reproductive cycles. We examined whether these techniques could provide this much needed information for a monomorphic anuran with an unconfirmed mating season in the wild. We analysed urinary estrone conjugate, testosterone, and progesterone metabolites to infer the time of breeding and to identify sex in the endangered Maud Island frog, Leiopelma pakeka. Testosterone metabolites in males and estrone and progesterone metabolites in females were at their peak during winter for both wild and captive frogs. These urinary metabolite patterns were consistent with the high proportion of females exhibiting enlarged ovarian follicles in winter months. Sex identification based on urinary estrone metabolite levels was 94% correct in this monomorphic species, in which the sexes overlap in snout-to-vent length (SVL) for over half of their adult size range and in which no other sexually dimorphic trait is known. The seasonal profiles imply unexpected winter or early spring breeding in L. pakeka. Overall, these results demonstrate use of urinary hormone metabolites for reproductive monitoring and sex identification in one of the world’s most threatened and evolutionarily distinct amphibians.

Increased Activation of the Alternative “Backdoor” Pathway in Patients with 21-Hydroxylase Deficiency: Evidence from Urinary Steroid Hormone Analysis

17-Hydroxyprogesterone (17-OHP) can be converted to dihydrotestosterone (DHT) via an alternative "backdoor" route that bypasses the conventional intermediates androstenedione and testosterone. In this backdoor pathway, 17-OHP is converted to 5α-pregnane-3α,17α-diol-20-one (pdiol), which is an excellent substrate for the 17,20 lyase activity of CYP17A1 to produce androsterone. OBJECTIVE AND HYPOTHESES: The objective of this study was to obtain evidence for the presence of the backdoor pathway in patients with 21-hydroxylase deficiency (21-OHD). We compared urinary steroid hormone profiles determined by gas chromatography-mass spectrometry of 142 untreated 21-OHD patients (age range, 1 d to 25.4 yr; 51 males) with 138 control subjects. The activity of the backdoor pathway was assessed using the ratios of the urinary concentrations of pdiol to those of the metabolites of the classic Δ4 and Δ5 pathways. In contrast to etiocholanolone, which originates almost exclusively from the classic pathways, androsterone may be derived additionally from the backdoor pathway. Therefore, the androsterone to etiocholanolone ratio can be used as an indicator for the presence of the backdoor pathway. Untreated 21-OHD subjects showed increased urinary ratios of pdiol to the Δ4 and Δ5 pathway metabolites and a higher androsterone to etiocholanolone ratio. The elevated ratios of pdiol to the Δ4 and Δ5 pathway metabolites as well as the higher androsterone to etiocholanolone ratio in patients with 21-OHD indicate postnatal activity of the backdoor pathway with maximum activity during early infancy. Our data provide new insights into the pathophysiology of androgen biosynthesis of 21-OHD.

Urinary hormone analysis assists reproductive monitoring and sex identification of bell frogs ( Litoria raniformis)

With the world currently facing a global amphibian extinction crisis, the development of techniques to help meet the needs of conservation managers and researchers studying the reproductive biology of amphibians is needed. Here, we developed enzyme immunoassays to measure estrone, testosterone, and progesterone hormone metabolites in the urine of Litoria raniformis, the southern bell frog. Concentrations of urinary estrone, testosterone, and progesterone increased during the breeding season for females (P < 0.05). Concentrations of urinary testosterone and progesterone increased for males during the breeding season compared with that for months where no reproductive behaviors were observed (P < 0.05). Furthermore, urinary estrone concentrations proved to be a reliable sexing tool for adult frogs, with no overlap between the sexes in 98% of cases, regardless of season. There was no difference in estrone (P = 0.204) or testosterone (P = 0.485) metabolite concentrations between samples taken immediately upon capture and those taken 12 to 24 h later from the same individual. Progesterone metabolite concentrations were lower on Day 2 than upon collection (P = 0.004). This is the first study to show that urinary hormone analysis can be a useful technique for reproductive monitoring in an amphibian. Additionally, hormone metabolite measures offer promise as sex identification tools for monomorphic species and for those whose secondary sex characteristics are visible only during the breeding season.

Evaluating the utility of an accelerometer and urinary hormone analysis as indicators of estrus in a Zoo‐housed koala (Phascolarctos cinereus)

Induced ovulators, such as the koala, do not always have overt signs of estrus, which makes pairing these animals for breeding purposes difficult to achieve in a zoo setting. This study examined the possibility of using alternative methods to behavioral sampling and weight fluctuations for monitoring estrus in a female koala of reproductive age. We attempt to gain an improved understanding of koala estrus and enhance our ability to detect it by combining a noninvasive technique for hormone analysis with a newer method for recording activity level. The findings suggest that activity levels were accurately measured using an accelerometer and increased activity was associated with one estrus behavior, bellowing. Weekly weight declines occur with increases in activity registered from the accelerometer. We also validated an assay to detect progesterone and estradiol in female koala urine in an attempt to detect an estrus spike. However, neither urinary concentrations of estradiol nor progesterone was associated with behavioral cues of estrus in our subject. We suggest that increased female activity, bellowing, and weight loss all indicate a pro-estrus state that requires proper male stimulation in order to achieve a full estrus display.

Urinary steroid hormone analysis of ovarian cycles and pregnancy in mandrills (Mandrillus sphinx) indicate that menses, copulatory behavior, sexual swellings and reproductive condition are associated with changing estrone conjugates (E1C) and pregnanediol‐3‐glucuronide (PdG)

The objective of this study was to determine if sexual swellings in mandrills (Mandrillus sphinx) are a reflection of reproductive endocrine state. Urine samples were assayed using an enzyme immunoassay measuring pregnanediol-3-glucuronide (PdG) and estrone conjugates (E(1)C). Hormone patterns of ovarian cycles, pregnancy and lactation were characterized and compared with sexual swellings and copulations relative to menses and peak E(1)C. Cycle lengths averaging 28.7 days and pregnancy length of 181 days determined by hormonal and sexual swelling measures were similar to those reported in other Old World primate species. First day of copulation was observed during rising E(1)C concentrations and preceded observations of peak swelling by 1-2 days. Observations of peak sexual swellings occurred at or on the day after peak E(1)C and decreased following the ovulatory increase in PdG. Observations of menses and sexual swellings are a useful method to track mandrill ovarian cycles and can assist zoos in determining the reproductive state of females in their collections. Zoo Biol 27:320-330, 2008. (c) 2008 Wiley-Liss, Inc.

Hormonanalysen im Urin

Urinary hormone analysis is applied to detect an altered steroid hormone metabolism, an elevated production of biogenic amines and to non-invasively determine the protein hormone human beta-choriogonadotropin indicating a pregnancy. Occasionally, these determinations need to be complemented by plasma- or serum hormone analysis. Clinical data including current drug therapy and urinary creatinine as reference are required to interpret any urine analysis. Diseases to be investigated by steroid hormone analysis are excess production of a typical or atypical mineralocorticoid active steroid hormones, the hormonal activity of adrenal or ovarian tumors, acne of unknown origin, hirsutism, a PCO-, an adrenogenital or a suspected Cushing syndrome. Biogenic amines should be determined in suspected secondary or refractory arterial hypertension, in case of pheochromocytoma- or paraganglioma-associated symptoms or if a serotonin-producing tumor is suspected. In children genetically determined diseases are the primary background to perform an analysis.

Reproductive monitoring of free-ranging female mountain gorillas by urinary hormone analysis.

Frequent urine samples from three habituated free-ranging adult female mountain gorillas were collected during a four-week period in order to characterize, by analysis of urinary hormones, their ovarian cycle, and to diagnose pregnancy. A conceptive cycle and two cycles that did not result in full term pregnancy were monitored. Behavioral and physiological observations of mating and labial swellings were correlated to urinary hormone results. Mating occurred on days with elevated estrogens. Labial swelling was maximum near or at peak estrogen levels. Pregnancy was diagnosed at the field site by measurement of chorionic gonadotropin using commercially available human pregnancy detection kits. In one pregnancy, gestation length was 254 days. The pregnanediol glucuronide concentrations of the mountain gorilla did not change during the nonconceptive cycle. During the conceptive cycle, the concentrations did not increase until the approximate time of implantation. This finding suggests marked differences in steroid production or metabolism between the mountain gorilla and the captive lowland gorilla.

Assessment of female reproductive status in captive-housed Hanuman langurs (Presbytis entellus) by measurement of urinary and fecal steroid excretion patterns.

The study reports on the use of urinary and fecal hormone measurements for monitoring female reproductive status in captive-housed Hanuman langurs (Presbytis entellus). Matched urine and fecal samples collected throughout 7 complete menstrual cycles of two females, and during part of one pregnancy in a third female were analyzed. Estrone conjugates (E1C) and immunoreactive pregnanediol glucuronide (PdG) in urine and immunoreactive estradiol (E2), progesterone (P4), pregnanediol (Pd) and 20α-hydroxyprogesterone (20αOHP) in feces were measured by enzymeimmunoassay. E1C and PdG in urine were excreted in a cyclic pattern with E1C levels increasing 3- to 4-fold during the follicular phase to reach preovulatory peak values 2 days before a defined rise in PdG concentrations. Cycle lengths ranged between 20 and 34 days comprising a variable follicular phase of 7-21 days and a more consistent luteal phase of 12-14 days. High pressure liquid chromatography (HPLC) analysis of fecal extracts confirmed the presence of all fecal hormones measured, but indicated large amounts of additional immunoreactivity in the three progestin assays. The patterns of excretion of fecal E2 and all three fecal progestins corresponded well with those of steroid metabolites in urine in showing a clear and well defined follicular phase E2 rise followed by a luteal phase progestin increase. Measurement of 20αOHP immunoreactivity revealed the most stable baseline and the highest follicular/luteal phase differential. Levels of all hormones were clearly elevated during pregnancy although urinary E1C and PdG showed a more pronounced increase compared to fecal metabolites. The results indicate that urinary and fecal hormone analysis can be applied to noninvasive monitoring of reproductive status in the Hanuman langur. © 1995 Wiley-Liss, Inc.

Equine immunocontraception using porcine zona pellucida: A new method for remote delivery and characterization of the immune response

Equine immunocontraception using porcine zona pellucida: A new method for remote delivery and characterization of the immune response

Breeding orangutans at Perth Zoo: Twenty years of appropriate husbandry

AbstractThe Perth Zoo orang‐utan husbandry strategy, which has been the foundation of 20 years of success in breeding, health, and longevity, is based on respect for the solitary arboreal nature of the species. The orang‐utans are housed singly at night and fed only in their night dens. Diets are based on the weight and energy expenditure of the individuals. During the day they are housed singly or in very small groups that correspond in size and structure to wild population units. Adult males and females are only introduced for breeding purposes. Large climbing frames, which permit visual and vocal contact, dominate the day enclosures. A wide variety of manipulable objects are provided daily to enhance activity levels.Since 1968 there have been 21 births, 14 surviving infants, and no post‐infancy deaths. All the mature animals are fertile. The major health problems of captive orang‐utans and the most common causes of adult mortality have been absent from the colony. The husbandry method used has made it possible to record detailed reproductive data including complete menstrual cycle records for 8 females since 1981, the observation and quantification of cyclic proceptivity for all mature females, the calculation of 19 gestation periods, and the measurement of post‐partum amenorrhea and interbirth intervals. Urinary hormone analysis is used regularly to confirm ovulation. A long‐term study of the hormones mediating the development of secondary sexual characteristics in males was started in 1988.Captive orang‐utans do not have a fertility problem, but the captive population is threatened by the unacceptably high mortality rate in young adults. This appears to be related to inappropriate husbandry methods. The most neglected aspect of orang‐utan husbandry is natural social group size and structure.

Fertility assessment: new methods and implications for endangered species

Practical and reliable methods for assessing reproductive status are essential for any work designed to enhance fertility in captive endangered species. This paper described recent advances in the development and application of such methods and their implications for the management of captive breeding. Since non-invasive procedures are necessary when working with animals that are easily stressed, urinary hormone analysis provides the most feasible approach to long term studies on most endangered species. Simplified hormone assay methods have been developed and applied to the detection of ovulation and pregnancy in a variety of animals, including primates, the Giant Panda and ungulates. The possibility now exists for extending the application of these methods to studies in the field. Finally, the ability to monitor reproductive status in endangered animals provides the basis for future attempts to accelerate captive breeding by artificial means.

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.

Reproductive studies unit of the New York Zoological Society

AbstractThe Reproductive Studies Unit of the New York Zoological Society addresses several areas of exotic animal reproduction. These include semen collection and freezing, embryo transfer in the gaur, study of normal reproductive anatomy, urinary hormone analysis of cycling and pregnant females, and postmortem collection and freezing of oocytes. The primary goal is development of exotic species gene banks.