The use of a urinary estrone conjugates assay for detection of optimal mating time in the cynomolgus macaque (Macaca fascicularis)

Forty-four female cynomolgus macaques (Macaca fascicularis) were examined to determine the optimum fertile period for mating. Daily urinary estrone conjugates (E1C) were measured, beginning on day 7 of the menstrual cycle, until a 1.5-gold E1C rise above the baseline was detected. The females were bred the next morning. Pregnancies were verified in all animals at day 18 postbreeding, and/or on day 25 postbreeding. Serum progesterone levels were used to correlate the relationship between ovulation and the E1C peak. Forty-four of the 57 cycles indicated a urinary E1C peak between days 10-15 of the menstrual cycle; this peak occurred on the day following the initial 1.5-fold to twofold rise in 90% of the cycles. A single 2-hr mating period the day before, the day of, or the day after the E1C peak resulted in conception in 17 of 44 (38.6%) animals.     

Is Cortisol Excretion Independent of Menstrual Cycle Day? A Longitudinal Evaluation of First Morning Urinary Specimens

Background

Cortisol is frequently used as a marker of physiologic stress levels. Using cortisol for that purpose, however, requires a thorough understanding of its normal longitudinal variability. The current understanding of longitudinal variability of basal cortisol secretion in women is very limited. It is often assumed, for example, that basal cortisol profiles do not vary across the menstrual cycle. This is a critical assumption: if cortisol were to follow a time dependent pattern during the menstrual cycle, then ignoring this cyclic variation could lead to erroneous imputation of physiologic stress. Yet, the assumption that basal cortisol levels are stable across the menstrual cycle rests on partial and contradictory evidence. Here we conduct a thorough test of that assumption using data collected for up to a year from 25 women living in rural Guatemala.

Methodology

We apply a linear mixed model to describe longitudinal first morning urinary cortisol profiles, accounting for differences in both mean and standard deviation of cortisol among women. To that aim we evaluate the fit of two alternative models. The first model assumes that cortisol does not vary with menstrual cycle day. The second assumes that cortisol mean varies across the menstrual cycle. Menstrual cycles are aligned on ovulation day (day 0). Follicular days are assigned negative numbers and luteal days positive numbers. When we compared Models 1 and 2 restricting our analysis to days between −14 (follicular) and day 14 (luteal) then day of the menstrual cycle did not emerge as a predictor of urinary cortisol levels (p-value >0.05). Yet, when we extended our analyses beyond that central 28-day-period then day of the menstrual cycle become a statistically significant predictor of cortisol levels.

Significance

The observed trend suggests that studies including cycling women should account for day dependent variation in cortisol in cycles with long follicular and luteal phases.     

Estrogen and LH dynamics during the follicular phase of the estrous cycle in the Asian elephant

Pituitary and corpus luteum hormone patterns throughout the elephant estrous cycle have been well characterized. By contrast, analysis of follicular maturation by measurement of circulating estrogens has been uninformative. This study tested the ability of a urinary estradiol‐3‐glucuronide radioimmunoassay to noninvasively assess follicular development during the nonluteal phase of the elephant estrous cycle, and to determine the relationship between estrogen production and the “double LH surge.” Daily urine and serum samples were collected throughout seven estrous cycles from three Asian elephants, and urine was collected from an additional three females, for a total of 13 cycles. Serum was analyzed for luteinizing hormone (LH), and urine was analyzed for estrogens and progestins. Elephants exhibited a typical LH pattern, with an anovulatory LH (anLH) surge occurring approximately 21 days before the ovulatory LH (ovLH) surge. The urinary estrogen pattern indicated the presence of two follicular waves during the nonluteal phase. The first wave (anovulatory) began 5 days before the anLH surge and reached a maximum concentration the day before the peak. Thereafter, urinary estrogens declined to baseline for 2 weeks before increasing again to peak concentrations on the day of the ovLH surge. Urinary progestins were baseline throughout most of the follicular phase, increasing 2–3 days before the ovLH surge and continuing into the luteal phase. These results support previous ultrasound observations that two waves of follicular growth occur during the nonluteal phase of the elephant estrous cycle. Each wave is associated with an increase in estrogen production that stimulates an LH surge. Thus, in contrast to serum analyses, urinary estrogen monitoring appears to be a reliable method for characterizing follicular activity in the elephant. Zoo Biol 22:443–454, 2003. © 2003 Wiley‐Liss, Inc.     

Long-term data on basic reproductive parameters and evaluation of endocrine,morphological, and behavioral measures for monitoring reproductive status in a group of semifree-ranging Tonkean macaques (Macaca tonkeana)

Cycle and gestation lengths, menstruation patterns, female genital swelling characteristics, and male-female consortship durations are reported in a semifree-ranging group of Tonkean macaques (Macaca tonkeana) studied over a 12 year period. In addition, profiles of urinary estrone conjugates (E1C) and immunoreactive pregnanediol glucuronide (PdG) throughout four complete menstrual cycles in two females and three full-term pregnancies are presented. Based on intermenstrual intervals, a mean cycle length of 37–41 days (n = 55 cycles in 10 females) was found. Gestation length averaged 173 days (n = 27 pregnancies in eight females). Measurement of PdG immunoreactivity in urine revealed a cyclic pattern with a 5–15-fold increase between follicular and luteal phase concentrations, suggesting that PdG is a reliable indicator of ovarian cyclicity and luteal function. In contrast to PdG, E1C excretion showed no clear pattern throughout the cycle; however, highest values of E1C were usually found shortly before the onset of the luteal phase PdG rise at the presumed time of ovulation. Levels of both hormones were elevated during the first half of gestation and showed a marked increase throughout the second half, with maximum E1C concentrations being up to 100-fold higher than nonpregnant levels. Consortships by the male and occurrence of female genital swelling were long lasting (on average 5–10 days and 13 days, respectively) and were restricted to the follicular phase of the cycle. The day of maximal swelling and day of detumescence as well as the end of male consortship were closely associated with the periovulatory period. Swellings and consortships were longer following lactational ammenorhea than for subsequent cycles. The evolutionary significance of the cyclical changes undergone by females upon their relations with males is discussed. © 1996 Wiley-Liss, Inc.     

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.     

Cytologic, hormonal, and ultrasonographic correlates of the menstrual cycle of the New World monkey Cebus apella

Few reports on the reproductive physiology of Cebus apella have been published. In this study we characterized menstrual cycle events by means of vaginal cytology, ultrasonography (US), and hormonal measurements in serum during three consecutive cycles in 10 females, and assessed the probability that ovulation would occur in the same ovary in consecutive cycles in 18 females. The lengths and phases of the cycles were determined according to vaginal cytology. Taking the first day of endometrial bleeding as the first day of the cycle, the mean cycle length +/- SEM was 19.5+/-0.4 days, with follicular and luteal phases lasting 8.2+/-0.2 and 11.3+/-0.4 days, respectively. The follicular phase included menstruation and the periovulatory period, which was characterized by the presence of a large number of superficial eosinophilic cells in the vaginal smear. The myometrium, endometrium, and ovaries were clearly distinguished on US examination. During each menstrual cycle a single follicle was recruited at random from either ovary. The follicle grew from 3 mm to a maximum diameter of 8-9 mm over the course of 8 days, in association with increasing estradiol (E(2)) serum levels (from 489+/-41 to 1600+/-92 pmol/L). At ovulation, the mean diameter of the dominant follicle usually decreased by >20%, 1 day after the maximum E(2) level was reached. Ovulation was associated with an abrupt fall in E(2), a decreased number of eosinophilic cells, the presence of leukocytes and intermediate cells in the vaginal smear, and a progressive increase in progesterone (P) levels that reached a maximum of 892+/-65 nmol/L on days 3-6 of the luteal phase. The menstrual cycle of Cebus apella differs in several temporal and quantitative aspects from that in humans and Old World primates, but it exhibits the same correlations between ovarian endocrine and morphologic parameters.     

Menstrual Phase Response to Nocturnal Light

The aims of the study were to test whether nocturnal white light can normalize menstrual cycles in oligomenorrheic women, and whether the phase of the menstrual cycle in which light is given is important for the shortening effect. Twenty-five women with long menstrual cycles (35.9–53.4 days on average) were treated for 1–3 cycles, each of which was preceded and followed by at least two untreated cycles. Treatments were 100 watt bedside lights administered for 5 consecutive nights. They centered at three different phases of the menstrual cycle: 6–7th, 14–17th or 23–25th days of the treated cycle (early, middle or late treatment, respectively). On average, the treatment cycle lengths were modestly, but significantly reduced compared to the duration of baseline cycles (more than 11 %). The difference in the effects of the early, middle and late treatment was not significant. However, if middle or late treatments were administered in the latter half of the interval between the menstrual cycle onset and probable time of ovulation, reductions of the treated cycle length were substantial (more than 20 %, resulting in cycles less than 33 days on average; p < 0.001). Other treatments produced only weak (up to 7 %), if any, cycle reductions. Moreover, we found a strong correlation (p < 0.001) between the duration of baseline cycle and differential effect of middle treatment (compared to early or late treatment). Middle treatments reduced treated cycle duration to the normal range in the subjects with shorter mean baseline cycles (<42 days), while in the subjects with longer duration of baseline cycle the shortening effect was produced by late treatments (p = 0.005 and p = 0.001, respectively). The results support the suggestion that a bedside lamp used on nights prior to ovulation can cause reduction of long menstrual cycles.     

The Sulawesi Crested Black Macaque (Macaca nigra) menstrual cycle: changes in perineal tumescence and serum estradiol, progesterone, follicle-stimulating hormone, and luteinizing hormone levels.

Events in the normal menstrual cycle of the endangered Sulawesi Crested Black Macaque (Macaca nigra) were characterized. Daily blood samples were obtained during 10 menstrual cycles from five M. nigra demonstrating regular cycles. The amount of perineal tumescence was scored daily. Serum levels of estradiol and progesterone were determined by RIA, serum LH levels were determined by the mouse Leydig cell bioassay, and serum FSH levels were determined by the rat granulosa cell aromatase bioassay. Cycle length was 39.8 +/- 1.0 days (mean +/- SEM) with an LH surge occurring 25 +/- 1.5 days from the onset of menses. After menses, both LH and estradiol were initially depressed, with estradiol first exceeding 50 pg/ml 8 days before the LH surge. In five cycles, peak estradiol levels (340 +/- 44 pg/ml) occurred on the day of the LH surge (637 +/- 58 ng/ml) and in the other five cycles, on the day before the LH surge. There was a broad increase of FSH in midcycle without a well-defined surge corresponding to the LH surge. Progesterone began increasing on the day of the LH surge and reached peak levels (6.8 +/- 0.96 ng/ml) 8 days later. Maximal perineal tumescence was generally associated with the time of the LH surge, but variation between animals made it impossible to predict accurately the day of the LH surge by perineal tumescence scores alone.     

Reproduction in captive Taiwan macaques (Macaca cyclopis) in comparison to other common macaque species

Twenty-five years of reproductive and birth data from a Macaca cyclopis colony at the New England Regional Primate Research Center (NERPRC) indicate that these animals are seasonal breeders with a birth peak between February and May. Interbirth intervals have a mean of 587.5 ± 45.6 days. The age at first reproduction for females is 5.08 ±.357 years and for males 5.01 ± .94. The normal menstrual cycle length ranges from 24–31 (mean 27.9 ± .506) days, with “offseason” long cycles ranging from 43–69 days. For 10 females studied prospectively, the sex skin color cycle ranged from 23–34 days and exhibited no long cycles. The peak estradiol value was 1.35 nmol/L on day 11 (range 10–12) of the menstrual cycle. Estradiol values ranged from 0.7–2.0 nmol/L. The peak progesterone value was 23 nmol/L on day 21 (17–23). Progesterone levels ranged from 10–42 nmol/L. The observed lower fecundability of M. cylopis in a timed mating program is consistent with later ages at first reproduction and longer interbirth compared to the other species in breeding colonies at NERPRC. © 1995 Wiley-Liss, Inc.     

Periovulatory time courses of plasma estradiol and progesterone in the Japanese monkey (Macaca fuscata)

Periovulatory time courses of plasma estradiol and progesterone were determined in 21 menstrual cycles of 20 Japanese monkeys. Both steroids were measured by radioimmunoassay. Ovulation was detected by serial laparoscopic observations of the ovaries. Three of the 21 cycles were anovulatory cycles. In the remaining 18 ovulatory cycles, a preovulatory estradiol peak occurred on day 12.2±1.4 (range 10–15) of the menstrual cycle. The estradiol concentration at the peak was 431±199 (range 210–930) pg/ml. The time interval between the estradiol peak and ovulation was within 48 hr; the shortest interval was 10–13 hr and the longest 32–48 hr. Although the progesterone levels began to increase slightly (0.6–1.4 ng/ml) before ovulation, they did not show a continuous increase but decreased once before ovulation. The increase in progesterone with development of the corpus luteum after ovulation was very gradual during the first 2 days after ovulation. Subsequently, in 13 of 18 ovulatory cycles the progesterone levels rose rapidly and reached a maximum, 4.0±1.2 (range 2.3–5.7) ng/ml, 4–8 days after ovulation. In 5 of the 18 cycles, the progesterone levels did not rise at all or did not exceed 2.0 ng/ml even if they showed more or less an increase. In the 5 cycles, the length of the luteal phase was 8.2±1.6 (range 6–10) days, which was significantly shorter than that of the former 13 cycles with 14.0±1.1 (range 13–16) days.     

Menstrual cycle characteristics and predictability of ovulation of Bhutia women in Sikkim, India

Although a woman's menstrual history can have significant implications for health outcomes, few studies have examined menstrual cycle variability in non-western, non-clinically based populations. This study presents menstrual cycle characteristics from Bhutia women living in Gangtok, Sikkim, India. The Bhutia are one of two indigenous populations residing in this small, northeastern state of India. A total of 1067 cycles were recorded by 200 Bhutia women over the course of 12 months. Mean cycle length in this population was similar to reported mean cycle lengths for populations in the U.S (30 days vs. 28 days). Menstrual cycles in this sample were highly variable with most women experiencing more than one short or long menstrual cycle. The frequency of irregular menstrual cycles experienced by individuals also varied significantly by season. A body mass index (BMI) above or below the WHO defined normal range was associated with higher rates of irregular cycles. Leutenizing hormone (LH) and follicle stimulating hormone (FSH) levels were also determined from urine samples collected just before mid-cycle, based on median cycle lengths. Although menstrual cycles in this sample were highly variable, median cycle length was still useful in predicting timing of the pre-ovulatory hormone surges of LH and FSH. Frequency of irregular cycles did impact the successful capture of the LH and FSH peak values.     

Correlation of perineal detumescence and ovulation in the pigtail macaque (Macaca nemestrina).

A study was made of the relationship between ovulation and perineal detumescence in the pigtail macaque (Macaca nemestrina). Daily laparoscopic examation of both ovaries was made beginning on the 9th day of the menstrual cycle. The appearance of the ovaries was recorded and photographs were taken of the perineum and ovaries. This procedure was repeated daily until the 5th day after ovulation or until the 20th day of the menstrual cycle for presumed anovulatory cycles. The time at which ovulation occurred was correlated with first signs of detumescence, both events occurring within a 24-hour period.     

Four-year study of controlled timed breeding of rhesus monkeys (Macaca mulatta)

As part of the timed breeding colony at Tulane National Primate Research Center, exogenous progesterone administration (5 mg/day for 10 days) has been used to select conception dates by inducing artificial luteal phases in female rhesus monkeys. A retrospective analysis of data obtained during four breeding seasons (1998-2001) revealed that conceptions occurred an average of 18 days after the last administration of progesterone. The age of the female to be bred, previous pregnancy history, and timing of breeding during the breeding season were determined to be critical factors in the success of the procedure. The benefit of this method of timed breeding is that it does not require tracking of menstrual cycles, which can be labor-intensive and requires that animals be monitored several months in advance of breeding to determine each female's individual cycle length. It also provided an efficient use of breeding-age males.     

Pattern of menstrual cycle length in South Indian women: A prospective study

Abstract

Data on 8,308 menstrual cycles from 1,740 South Indian women prospectively recorded were analyzed to identify the effect of age on menstrual cycle length. The distribution was skewed to the right with the mean (SD) cycle length of 31.8 (6.7) days. The range of 25–40 days constituted 10–82 per cent of menstrual cycle lengths. In no age group did 28‐day cycles occur in more than 9 per cent of women. Variability as measured by the standard deviation was high among those below 19 years of age, stabilized during 25–39 years, and then increased in women aged 40 years or more. The findings are discussed in the light of other studies and possible implications in fertility control programs.     

Temporal and endocrine sequelae of aspirating follicular contents in rhesus monkeys

Aspiration of ovarian follicular contents in humans is a well-established procedure used to obtain oocytes for fertilization in vitro (IVF). However, the effects of aspiration on the menstrual cycle and resulting luteal function have been incompletely characterized. The present study was designed to investigate alterations in the temporal and endocrine characteristics of menstrual cycles following aspiration of contents of the dominant preovulatory follicle (DF) on day 10 of the cycle in normal rhesus monkeys. When aspiration was performed prior to the preovulatory surge of luteinizing hormone (LH), cycle length was extended to 38.6 ± 8.6 [15] (x days ± SD, [n]), as compared to 29.5 ± 5.7 [8] days when the surge occurred before the time of aspiration. Mean and maximal amounts of progesterone (P) in the luteal phase and the number of days in which P-values were > 1 ng/ml were significantly greater when aspiration was performed prior to the surge of LH than for aspiration after this event. Laparoscopic observations made in the midluteal phase in animals of the former group demonstrated that the corpus luteum (CL was derived from a follicle other than the original DF which had been aspirated on day 10 of the menstrual cycle; observations in the latter group of animals indicated that the CL was derived from the DF.     

Some characteristics of the normal menstrual cycle of the bonnet monkey (M. radiata)

Sixty-two menstrual cycles were observed in five bonnet monkeys over a period of 12 months. The menstrual cycles ranged in length from 22 to 35 days, although cycles of from 26 to 29 days were most frequent. Menstrual flow occurred for two to seven days, the flow lasting for three to four days in the majority (82%) of the cycles. Serum estradiol-17β (E₂) and progesterone (P) were measured by radioimmunoassay in two consecutive cycles in each monkey and correlated with changes in the quantity of cervical mucus secretion and spinnbarkeit. The day of the estradiol peak was considered as day 0. The E₂ levels began to rise one to three days before the peak, returning to baseline levels within two days and thereafter remained at low levels. The serum P rose 2 days after the E₂ peak, and maximum level was attained at 10 to 12 days after the E₂ peak. The quantity of cervical mucus increased proportionately with estrogen. However, the quantity remained at maximum levels even after the decline of the serum E₂ level. Similar trends were observed with the spinnbarkeit.     

Effect of dexamethsone on luteinizing hormone and follicle stimulating hormone responses to LHRH and to clomiphene in the follicular phase of women with normal menstrual cycles.

In an attempt to elucidate the mechanism of suppressive action of glucocorticoids on the hypothalamo-pituitary-ovarian axis, we studied the effects of short-term high dose dexamethasone administration of the LH and FSH responses to LHRH and to clomiphene in healthy women with normal menstrual cycles. Seven women, 21--35 years of age, received 100 micrograms of LHRH i.v. on day 6 of two consecutive menstrual cycles, once with and once without pre-treatment with dexamethasone 2 mg orally every 6 hrs. on days 2 through 5 of the menstrual cycle. Seven other women (ages 21--35 years) received clomiphene citrate 100 mg on days 2 through 5 of their menstrual cycle, once with and once without simultaneous administration of dexamethasone 2 mg orally every 6 h. The administration of dexamethasone suppressed baseline serum levels of LH and FSH and blunted LH and FSH response to both LHRH and clomiphene. The results indicate that short-term administration of pharmacological doses of glucocorticoids suppress the secretion of LH and FSH by a direct effect on the anterior pituitary and possibly by an effect at the hypothalamic level with inhibition of the release of LHRH.     

Disturbance of the Duration in the Menstrual Cycle under the Influence of Tight Clothing

We investigated the influence of skin pressure by clothing on the duration of menstrual cycle with 33 young adult women. The average age was 19.9 ± 2.1 years (mean ± SD), stature 159.5 ± 5.6cm and body mass 50.9 ± 5.5kg. Thirty-three women participated as subjects. They wore their usual clothing including foundation garments, panty stocking, pants or skirt and T-shirt or blouse and cardigan for the first 4 months from December to March ('Tight 1'). For the second 4 months from April to July, the women wore loose clothes, i.e., they did not wear foundation garments at home. Skirt and jeans were worn loosely ('Loose'). For the last 4 months, from August to November, they wore their clothes as tightly as possible, compared to 'Tight 1' ('Tight 2'). Each participant marked the first day of the occurrence of menses in the pocket diary throughout the year. The main results were summarized as follows: 1) The average duration of the menstrual cycle was 44.2 ± 14.9 days (mean ±) in 'Tight 1', 30.4 ± 3.0 days in 'Loose' and 47.4 ± 22.7 days in 'Tight 2'. 2) The number of months when the menses did not occur was 38 for 'Tight 1', 6 for 'Loose' and 40 for 'Tight 2'. 3). The number of participants who had a duration of the menstrual cycles for more than 40 days, was 25 participants for 'Tight', 10 for 'Loose' and 29 for 'Tight'. It can be concluded that skin pressure by clothing could disturb the duration in the menstrual cycle.     

Disturbance of the Duration in the Menstrual Cycle under the Influence of Tight Clothing

We investigated the influence of skin pressure by clothing on the duration of menstrual cycle with 33 young adult women. The average age was 19.9 ± 2.1 years (mean ± SD), stature 159.5 ± 5.6cm and body mass 50.9 ± 5.5kg. Thirty-three women participated as subjects. They wore their usual clothing including foundation garments, panty stocking, pants or skirt and T-shirt or blouse and cardigan for the first 4 months from December to March (‘Tight 1’). For the second 4 months from April to July, the women wore loose clothes, i.e., they did not wear foundation garments at home. Skirt and jeans were worn loosely (‘Loose’). For the last 4 months, from August to November, they wore their clothes as tightly as possible, compared to ‘Tight 1’ (‘Tight 2’). Each participant marked the first day of the occurrence of menses in the pocket diary throughout the year. The main results were summarized as follows: 1) The average duration of the menstrual cycle was 44.2 ± 14.9 days (mean ±) in ‘Tight 1’, 30.4 ± 3.0 days in ‘Loose’ and 47.4 ± 22.7 days in ‘Tight 2’. 2) The number of months when the menses did not occur was 38 for ‘Tight 1’, 6 for ‘Loose’ and 40 for ‘Tight 2’. 3). The number of participants who had a duration of the menstrual cycles for more than 40 days, was 25 participants for ‘Tight’, 10 for ‘Loose’ and 29 for ‘Tight’. It can be concluded that skin pressure by clothing could disturb the duration in the menstrual cycle.     

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.