Monitoring fecal samples for estrogen excretion across the ovarian cycle in Goeldi's monkey (Callimico goeldii)

In captive Goeldi's monkeys, estrogen concentration was determined in fecal samples collected from 4 cycling/unmated females and 4 postpartum/mated females in order to ascertain the potential of fecal estrogen monitoring for providing basic information about reproductive status in this endangered Amazonian monkey. Subjects were fed an omnivorous diet and first-void feces were collected in the home cage at 1–3-day intervals for 30–50 days from the cycling females, and at 6–14-day intervals around the estimated time of the postpartum ovulation in each of the 4 mated females. Estimates for the duration of ovarian cycles (22–26 days) and the timing of ovulation were based on cyclic profiles of either blood progesterone or urinary pregnanediol-3α-glucuronide. Fecal estrogen values were normalized using these plasma or urinary profiles. HPLC analysis of estrogen from postpartum fecal samples demonstrated the presence of unconjugated estrone and estradiol-17β (“unconjugated estrogen”). Unconjugated estrogen was extracted and its fecal concentration estimated via EIA. The correlation (r) between plasma estrone conjugates and fecal unconjugated estrogen across nonconception ovarian cycles was 0.65 and measurement of the latter generated cyclic profiles. A range of 4–36-ng unconjugated estrogen g^?1 feces was identified for follicular phases of nonconception cycles. Fecal unconjugated estrogen first exceeded the concentration range of the follicular phase 2–5 days after ovulation; the range was 49–402 ng g^?1 feces in samples collected during the remainder of these nonconception luteal phases. Luteal phase concentrations were on average 10-fold higher than follicular phase concentrations. Each of the 4 mated females conceived at its postpartum ovulation; concentrations of fecal unconjugated estrogen excreted by 3 of these females demonstrated a marked postovulatory increase. This study demonstrates that fecal unconjugated estrogen can be applied to monitor ovarian cyclicity in Goeldi's monkey. © 1994 Wiley-Liss, Inc.     

Use of salivary steroid analyses to assess ovarian cycles in an Indian rhinoceros at the National Zoological Park

Saliva samples were collected from a female Indian rhinoceros (Rhinoceros unicornis) housed at the National Zoological Park, and ether-extracted for analyses of androgen, estrogen, and progestin metabolites to assess ovarian cycles. Analyses of both salivary androgens and estrogens were found to reliably monitor follicular activity. Although the temporal patterns of the two steroids were significantly correlated (r=0.62; P<0.05), they differed slightly. Salivary androgens increased earlier during the follicular phase, although both peaked at the same time in association with behavioral estrus. Based on salivary androgen profiles, the mean duration of the follicular phase was 11.4 days (ranged=7–15 days; n=17 cycles). Estrous cycle length, as measured by the time between consecutive androgen peaks, was 47.4±3.4 days (range=37–86 days). Salivary progestin measurements were effective for monitoring luteal function. The mean duration of the luteal phase was 15.5±1.5 days (range=10–23 days). In sum, assays were identified for measuring salivary steroids to assess ovarian function in Indian rhinoceroses. However, not all of the assay systems tested were effective, perhaps because of interfering matrix effects. Mate introductions in the Indian rhinoceros often require careful monitoring, and a technique for monitoring hormones in saliva could be an alternative to urine or fecal analyses for identifying estrus and timing breeding in this species. Such a technique would be particularly useful for situations in which it is difficult to collect uncontaminated urine and feces. Zoo Biol 23:501–512, 2004. © 2004 Wiley-Liss, Inc.     

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.     

Use of group-specific antibodies to detect fecal progesterone metabolites during the estrous cycle of cows

Progesterone is metabolized to pregnanediones and hydroxylated pregnanes prior to its fecal excretion. Therefore, use of progesterone antibodies underestimates the actual amount of fecal metabolites. To improve the methodology of noninvasive fecal progesterone metabolite analysis, enzymeimmunoassays (EIA) using group-specific antibodies against 5-reduced 20-oxo-pregnane-C3-conjugates were developed. Fecal and milk samples were collected at 1- to 2-d intervals during the morning and evening milking throughout 1 estrous cycle in dairy cows (n = 12). Six immunoreactive metabolites were detected in the feces with high performance liquid chromatography (HPLC), eluting as 5α- and 5β-reduced pregnanes containing a 20-oxo-group (20-oxo-pregnanes). Fecal samples of 3 cows were analyzed by 3 EIAs using antibodies against 4-pregnene-6α-ol-3,20-dione 6HS:BSA (6HS-progesterone), 5α-pregnane-3β-ol-20-one 3HS:BSA and 5β-pregnane-3β-ol-20-one 3HS:BSA, respectively. The follicular and luteal phases were identifiable with each EIA. Luteal phase values and the differences between mean follicular (Days 0 to 2 and 19 to 21) and luteal phase (Days 10 to 16) values obtained with the 5-pregnane EIAs were 3- to 4-fold higher than with the 6HS-progesterone EIA. Since results with the former 2 EIAs were almost identical, the remaining samples were only analyzed by the EIA for 5β-pregnane-3α-ol-20-one. Fecal 20-oxo-pregnane concentrations were parallel to milk progesterone values, but had a lag time of about 0.5 d; the coefficient of correlation (P < 0.001) was 0.76 (y = 155.2 × + 37.2). Fecal 20-oxo-pregnane concentrations during the follicular and luteal phase were 39.5 ± 2.2 and 341 ± 15.2 ng/g feces, respectively. In conclusion, fecal 20-oxo-pregnanes are significantly correlated to milk progesterone concentrations. They consist of several metabolites and compared to a 6HS-progesterone antibody, their evaluation was improved using antibodies against 5-reduced pregnanes.     

Comparison of three protocols for superovulation of brown brocket deer (Mazama gouazoubira)

This study aims to evaluate the ovulation rate and the presence of functional corpora lutea after treatment by three different protocols designed to cause superovulation in brown brocket deer. Six female received an intravaginal device containing 0.33 g of progesterone (CIDR®) for 8 days, followed by 0.5 mg injection of estradiol benzoate at the time of insertion and 265 µg of cloprostenol at the time of removal. Afterwards, the hinds were divided into three groups (n = 2): Treatment A received injection of 600 IU eCG on Day 4 after CIDR® insertion; Treatment B received injection of 300 IU eCG at the same time; and Treatment C received injection of 250 IU FSH dissolved in PVP, also on Day 4 post‐insertion. The treatments were crossed over with 44–48 day intervals after CIDR® removal, such that all the deer were submitted to all three treatments. The mean ovulation rate (Treatment A = 3.40 ± 0.68, Treatment B = 1.40 ± 0.24, Treatment C = 0.80 ± 0.49), total ovarian stimulation (Treatment A = 4.80 ± 1.02, Treatment B = 1.80 ± 0.37, Treatment C = 1.40 ± 0.60), and mean CL diameter (Treatment A = 7.33 ± 0.76 mm, Treatment B = 3.94 ± 0.19 mm, Treatment C = 2.18 ± 0.49 mm) in Treatment A were significantly higher than the mean ovulation rates, total ovarian stimulation, and mean CL diameter in Treatments B and C. The mean fecal progesterone metabolites at the luteal phase in Treatment A (6,277.94±2,232.47 ng/g feces) was significantly different from Treatment C (1,374.82±401.77 ng/g feces). Thus, although fertility was not evaluated directly, Treatment A proved capable of induce superovulation in the species Mazama gouazoubira, presenting the greatest mean ovulation rates, with the formation of functional corpora lutea. lutea. Zoo Biol 31:642‐655, 2012. © 2011 Wiley Periodicals, Inc.     

Ovarian cycle of southern brown howler monkey (<Emphasis Type="Italic">Alouatta guariba clamitans</Emphasis>) through fecal progestin measurement

The ovarian cycle in howler monkeys (genus Alouatta) has beean investigated through several biological parameters (ranging between 16.3 and 29.5 days); however, no data exist concerning the ovarian activity of the southern brown howler monkey (Alouatta guariba clamitans). This study aimed to describe the ovarian cycle of A. g. clamitans by profiling fecal progestin concentrations. Over 20 weeks, fecal samples of eight captive adult females of A. g. clamitans were collected. The collections were made at dawn, 5 days a week, and the samples were frozen immediately following collection. Next, they were dried, pulverized and hormonal metabolites were extracted to determine progestin concentrations by enzyme immunoassay. Of the 758 samples tested, the mean concentration of fecal progestins was 2866.40 ± 470.03 ng/g of dry feces, while the mean concentration at baseline was 814.47 ± 164.36 ng/g of dry feces. Among the eight females, one showed no ovarian cyclicity and three presented periods of probable absence of cyclicity and low progestin concentrations. A mean duration of 16 ± 0.52 days was observed for the 35 cycles studied. The interluteal phase lasted 4 ± 0.37 days on average, with a mean concentration of fecal progestins of 467.98 ± 29.12 ng/g of dry feces, while the luteal phase lasted 11 ± 0.50 days, with a mean concentration of 4283.27 ± 193.31 ng/g of dry feces. Besides describing the characteristics of the ovarian cycle, possible causes for the low concentrations of fecal progestins and periods of absence of cyclicity are also discussed.     

Assessment of ovarian function in the African elephant (Loxodonta africana) by measurement of 5α-reduced progesterone metabolites in serum and urine

We have previously shown that 5α-pregnane-3,20-dione (5αDHP), and 5α-pregnane-3-ol-20-one (5α-P-3-OH) are the major luteal and circulating progestins in the African elephant. Therefore, the aim of the present study was to determine (1) circulating levels and patterns of secretion of 5α-DHP in relation to progesterone (P4) throughout the ovarian cycle, (2) the presence and relative abundance of 5α-reduced progestins in urine and (3) whether their measurement in urine would provide a non-invasive method for monitoring luteal function. Urine samples were collected weekly throughout a total of 13 ovarian cycles from 5 females. In addition, matched blood samples were collected during 6 cycles from 2 of the 5 animals. All hormone measurement, were carried out by enzymeimmunoassay following extraction. Urine was hydrolyzed prior to extraction. Profiles of P4 and 5α-DHP in serum followed a similar cyclic pattern and both measurements were significantly correlated (r = 0.78–0.98, mean 0.89, P < 0.001). Concentrations of 5α-DHP were, however, 10–20 fold higher than those of P4. I addition, 5α-DHP measurements showed a more pronounced luteal phase increase than that of immunoreactive P4. HPLC co-chromatography confirmed the presence of large amounts of 5α-P-3-OH in urine as a single immunoreactive peak, whereas 5α-DHP was present in very low levels and measurable only as one of several immunoreactive substances. Measurements of urinary 5α-P-3-OH were significantly correlated to serum 5α-DHP measurements in each of the 6 cycles (r = 0.72–0.93, mean 0.81, P < 0.001), whereas correlation coefficients between urinary and serum 5α-DHP values were generally lower (r = 0.34–0.83, mean 0.69) and significant in only 4 of the 6 cycles. Accordingly, only urinary excretion of 5α-P-3-OH, but not of 0.15–0.20 μ/mg Cr in the follicular phase and 10-fold elevated levels (1.8–2.2 μg/mg Cr) in the luteal phase. Based on the intervals between successive luteal phase increases in urinary 5α-P-3-OH, a mean cycle length of 14.1 ± 1.8 weeks, comprising a follicular phase of 5.0 ± 0.9 weeks and a luteal phase of 9.1 ± 1.4 weeks was determined for the 13 cycles studied. The results indicate that measurements of 5α-P-3-OH in urine provide a reliable non-invasive method for monitoring luteal function in the African elephant. Zoo Biol 16:273–284, 1997. © 1997 Wiley-Liss, Inc.     

Reproductive gonadal steroidogenic activity in the fishing cat (Prionailurus viverrinus) assessed by fecal steroid analyses

Non-invasive fecal steroid analyses were used to characterize gonadal activity in the fishing cat (Prionailurus viverrinus). Estrogen, progestagen and androgen metabolites were quantified in fecal samples collected for 12 months from four males and 10 females housed at seven North American zoological institutions. Male reproductive hormone concentrations did not vary (P>0.05) among season, and estrogen cycles were observed year-round in females and averaged (±SEM) 19.9±1.0 days. Mean peak estrogen concentration during estrus (460.0±72.6ng/g feces) was five-fold higher than baseline (87.3±14.0ng/g feces). Five of seven females (71.4%) housed alone or with another female demonstrated spontaneous luteal activity (apparent ovulation without copulation), with mean progestagen concentration (20.3±4.7μg/g feces), increasing nearly five-fold above baseline (4.1±0.8μg/g feces). The non-pregnant luteal phase averaged 32.9±2.5 days (n=13). One female delivered kittens 70 days after natural mating with fecal progestagen concentrations averaging 51.2±5.2μg/g feces. Two additional females were administered exogenous gonadotropins (150IU eCG; 100IU hCG), which caused hyper-elevated concentrations of fecal estrogen and progestagen (plus ovulation). Results indicate that: (1) male and female fishing cats managed in North American zoos are reproductively active year round; (2) 71.4% of females experienced spontaneous ovulation; and (3) females are responsive to exogenous gonadotropins for ovulation induction, but a regimen that produces a normative ovarian steroidogenic response needs to be identified.     

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

A 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.     

Correlation of perineal swelling with serum ovarian hormone levels,vaginal cytology,and ovarian follicular development during the baboon reproductive cycle

Perineal swelling was correlated with changes in vaginal cytology and serum ovarian hormone levels and with development of the ovarian follicle. The average length of 66 menstrual cycles as seen in 12 mature baboons was 35.7±.66 days with a range of 25 to 47 days. Laparoscopic observations and photographic documentation of follicular development were made as early as 13 days prior to ovulation with the most rapid follicular maturation occurring 24 to 48 hours before ovulation. In 38.5% of the cycles ovulation occurred on the last day of maximal perineal tumescence with 26.9% of the ovulations occurring one day after initial detumescence. In 17.8% ovulations occurred two to five days prior to detumescence while the remaining 17.8% occurred two to three days following first observed detumescence. The mean progesterone level during the follicular phase was significantly less than that in the luteal phase, 1.6 and 6.7 ng/ml respectively (p<0.01). The level of estrogen detected during the periods of maximal tumescence was significantly higher (p<0.05) than that detected in the remainder of the cycle, 19.8 and 5.9 pg/ml respectively. Vaginal smears were stained, observed microscopically, and found to correlate with perineal swelling, hormone levels, and laparoscopic observations.     

Comparative analysis of gonadal and adrenal activity in the black and white rhinoceros in North America by noninvasive endocrine monitoring

Patterns of fecal reproductive steroid metabolites and adrenal corticoids were characterized for 12‐ to 24‐month periods in black (n = 10 male, 16 female) and white (n = 6 male, 13 female) rhinoceroses at 14 institutions. All black rhinoceros females exhibited at least some ovarian cyclicity on the basis of fecal progestogen analysis (range, 2–12 cycles/yr). However, cycles often were erratic, with many being shorter (<20 days; 18% of cycles) or longer (>32 days; 21%) than the average of 26.8 ± 0.5 days (n = 104 cycles). Five females exhibited periods of acyclicity of 2–10‐month duration that were unrelated to season. One complete and seven partial pregnancies were evaluated in the black rhinoceros. Fecal progestogens increased over luteal phase concentrations after 3 months of gestation. Females resumed cyclicity within 3 months postpartum, before calves were weaned (n = 5). Approximately half of white rhinoceros females (6 of 13) showed no evidence of ovarian cyclicity. Of the cycles observed, 5 were “short” (32.8 ± 1.2 days) and 24 were “long” (70.1 ± 1.6 days). Only two females cycled continuously throughout the study. One had both long (n = 9) and short (n = 2) cycles, whereas the other exhibited long cycles only (n = 5). Fecal estrogen excretion was variable, and profiles were not useful for characterizing follicular activity or diagnosing pregnancy in either species. Males of both species showed no evidence of seasonality on the basis of fecal androgen profiles. Androgen metabolite concentrations were higher (P < 0.05) in the black (27.6 ± 6.9 ng/g) than in the white (16.8 ± 3.1 ng/g) rhinoceros. An adrenocorticotropin hormone challenge in four black rhinoceros males demonstrated that the clearance rate of corticoid metabolites into feces was ～24 hours. Fecal corticoid concentrations did not differ between males and females, but overall means were higher in the black (41.8 ± 3.1 ng/g) than in the white (31.2 ± 1.7 ng/g) rhinoceros. In summary, fecal steroid analysis identified a number of differences in hormonal secretory dynamics between the black and white rhinoceros that may be related to differences in reproductive rates in captivity. Most black rhinoceros females exhibited some cyclic ovarian activity. In contrast, few white rhinoceroses demonstrated evidence of regular estrous cyclicity, and those females that were active had comparatively long cycles. Results also suggest that fecal corticoid concentrations reflect adrenal activity and may be species specific. Continued studies are needed to determine whether fecal corticoid measurements will be useful for understanding the cause of inconsistent gonadal activity in these two species. Because all but three (15.8%) of the white rhinoceroses evaluated in this study were less than 20 years of age compared to 73.1% (19 of 26) of the black rhinoceroses, the impact of age on reproductive and adrenal activity also needs to be evaluated further. Zoo Biol 20:463–486, 2001. © 2002 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.     

Bovine Steroid Hormone and SHBG Concentrations Postpartum and during the Oestrous Cycle

Changes in consecutive estimates of milk progesterone concentrations and serum steroid hormone and sex hormone-binding globulin (SHBG) concentrations in the postpartum period were examined in Finnish Ayrshire and Friesian dairy cows which were divided according to feeding into a hay group and a silage group. Milk progesterone concentrations rose above 10 nmol/1, indicating the start of ovarian luteal activity, slightly earlier in the silage group (28.4 ± 8.7 (S.D.) days, n = 19) than in the hay group (33.4 ± 10.3, n = 28) after calving. Likewise, the first normal oestrous cycles began slightly earlier in cows fed with silage. On the other hand, no differences in the beginning of ovarian luteal activity were observed between the breeds. Serum oestradiol-17β, oestrone, testosterone, 5α-dihydrotestosterone (5α-DHT), pregnenolone and progesterone concentrations were fairly unchanged during postpartum anoestrus after uterine involution and before ovarian cyclic activity. After first ovulation, considerable increases in milk and serum progesterone concentrations were observed. The increase was accompanied by elevations in serum pregnenolone and 5α-DHT concentrations. In the late luteal phase, progesterone, 5α-DHT and pregnenolone concentrations rapidly declined, leading to low hormone levels in pro-oestrus. Thereafter, serum pregenolone and 5α-DHT concentrations slightly increased during the follicular phase. On the other hand, oestradiol-17β concentrations were elevated in pro-oestrus and decreased after that, being lowest at met-oestrous. Serum testosterone concentrations appeared to be unchanged during postpartum anoestrus and over the oestrous cycle. Serum SHBG concentrations were unchanged during postpartum anoestrus and over the oestrous cycle, as well as in pregnant animals. The serum SHBG concentrations were about double those found in women with normal menstrual cycles, whereas oestradiol concentrations were much lower. At present, it cannot be explained how the biological effects of oestradiol become evident under such conditions.     

Effect of enucleation of the corpus luteum at different stages of the luteal phase of the human menstrual cycle on subsequent follicular development

To investigate the mechanism of suppression of follicular development during the luteal phase of the human menstrual cycle, the corpus luteum was enucleated surgically from 10 women at various times after ovulation. In the 24 h after CL enucleation there was an immediate and rapid fall in the concentration of oestradiol and progesterone and a temporary decline in the concentration of FSH and LH. Within 3 days, however, all 10 women showed evidence of renewed follicular activity as indicated by a progressive rise in the concentration of oestradiol. This rise was preceded by a rise in the concentration of FSH and LH, and ovulation, as indicated by a mid-cycle surge in LH and rise in the concentration of plasma progesterone, occurred 16-19 days after enucleation. There was no significant difference in the time to ovulation following enucleation at different times of the luteal phase. The post-operative follicular phase, measured from the time of enucleation, was 3 days longer than that observed pre-operatively from the first day of menstrual bleeding. In the follicular phase of post-operative cycles the concentration of FSH was higher and that of oestradiol lower than the corresponding values before surgery. These results indicate that the absence of healthy antral follicles in the luteal phase of the cycle is due to the inhibitory effects of the corpus luteum. The fact that, after CL enucleation, emergence of the dominant follicle was always preceded by a rise in the concentration of FSH and LH suggests that suppression of gonadotrophins by ovarian steroids secreted by the corpus luteum is responsible for the inhibition of follicular development during the luteal phase of the cycle.     

Suppression of ovulation in Nile Hippopotamus (Hippopotamus amphibious) using melengestrol acetate‐treated feed or high dose Depo‐Provera injection

Analysis of fecal progestogen profiles during Depo‐Provera injection (1,200 mg; DEPO, Pfizer Inc., New York, NY), melengestrol acetate (MGA) in feed (2 or 3 mg/head/day), and a combination treatment (DEPO+MGA) are presented for nine captive female Nile hippos housed at Disney's Animal Kingdom in Florida. All tested treatments reduced fecal progestogen elevations successfully to durations consistent with prevention of ovulation for a portion of the treatment period. Percentage of treatment months with suppression of luteal phases indicative of ovulation was maximal for high‐dose MGA (91.7±13.9%) and DEPO+MGA (91.7±20.4%), followed by DEPO injection alone (69.2±13.9%) and low‐dose MGA (57.6±33.2%). Both 1,200 mg DEPO and low‐dose MGA (2.0 mg/day) treatments were insufficient to prevent an apparent seasonal breakthrough of ovarian activity from June–August 2002. Although luteal phases were observed, no females conceived during those months. Overall, in 133.5 treatment months with females housed with an adult male, one female conceived during the transition period between treatments. After cessation of contraceptive treatment, average latency to first normal ovarian cycle was 80.6±19.5 days (range = 22–179 days). Up to 12 months post‐treatment, however, successive cycles were often irregular with evidence of short periods of anovulation and shortened luteal phases in all females monitored. In conclusion, high dose and combination treatments were most successful in preventing progestogen increases indicative of ovulation in hippos. Zoo Biol 26:259–274, 2007. © 2007 Wiley‐Liss, Inc.     

Behavioral and endocrine characteristics of the reproductive cycle in wild muriqui monkeys,Brachyteles arachnoides

The analysis of fecal ovarian steroids provides a powerful noninvasive method to obtain insights into ovulatory cycles, gestation length, and the timing of sexual interactions relative to the periovulatory period in wild primates. Techniques developed to collect and assay feces from free-ranging muriqui monkeys (Brachyteles arachnoides) for estradiol and progesterone yield the first explicit reproductive data on this species, and provide the first opportunity to evaluate the timing of observed copulations with muriqui ovarian cycles. Hormonal profiles from seven females indicate average cycle lengths of 21.0 ± 5.4 days (n=20). Females conceived after 3–6 ovulatory cycles. Gestation length averaged 216.4 ± 1.5 days for the five females for which conception cycles were sampled. Discrete copulation periods spanned an average of 2.1 ± 1.2 days (n=29), with intervals between these concentrated periods of copulations averaging 15.6 ± 6.7 days (n=20). There were no significant differences among females in cycle lengths, copulation period lengths, or copulation interval lengths. Ejaculation was visible following 71.8 ± 26.7% of copulations during the females' preovulatory periods. All females copulated outside the periovulatory period. The proportion of copulation days outside the periovulatory period was slightly greater (p=0.08) for primiparous females (64.8 ± 28.3%) than for multiparous females (28.7 ± 19.7%). Am. J. Primatol. 42:299–310, 1997. © 1997 Wiley-Liss, Inc.     

Laparoscopic observations of ovaries before and after ovulation in the Japanese monkey (Macaca fuscata)

Laparoscopic observations of ovaries before and after ovulation were made in 74 cycles in 39 Japanese monkeys (Macaca fuscata). Lengths of menstrual cycles, the follicular phase and the luteal phase in these animals were 25.3±2.8 days, 13.2±1.4 days, and 12.8±2.3 days, respectively. Ovarian morphology during the menstrual cycle in the Japanese monkey agreed in some features and disagreed in others, with findings inMacaca mulatta andMacaca fascicularis reported in the previous papers using laparotomy or laparoscopy. Follicular appearance before and after ovulation varied so considerably from ovary to ovary that it was hard to predict the accurate ovulation time or to estimate the precise age of corpus luteum by means of observations on the morphological appearance only. However, diagnosis of whether ovulation had already occurred or not, was possible when careful observations were made, although some ovaries without haemorrhagic and luteinized appearance after ovulation were very similar to pre-ovulatory follicle. Clear luteinization was detected without fail in many cases three days after ovulation. This study was supported by the Scientific Research Fund of the Ministry of Education of Japan (No. 856165).     

Reproductive steroid hormones and ovarian activity in felids of the Leopardus genus

Reproductive endocrine patterns were characterized in female ocelots (Leopardus pardalis; n = 3), tigrinas (Leopardus tigrinus; n = 2), and margays (Leopardus wiedii; n = 2) housed in captivity in southern Brazil. Females were maintained as singletons and exposed to natural fluctuations in photoperiod. Cyclic changes in ovarian steroids were monitored by analyzing estrogen and progestogen metabolites in fecal samples collected five times weekly for 14 to 18 months. Based on intervals between fecal estrogen peaks, mean (± SEM) duration of the estrous cycle was 18.4 ± 1.6 days for the ocelots (range, 7–31 days; n = 75 cycles), 16.7 ± 1.3 days for the tigrinas (range, 11–27 days; n = 23 cycles), and 17.6 ± 1.5 days for the margays (range, 11–25 days; n = 32 cycles). Fecal progestogen analyses combined with two laparoscopic observations of the ovaries confirmed that ocelots and tigrinas did not ovulate spontaneously. In contrast, non‐mating–induced luteal phases of 40.1 ± 6.3 days in duration (range, 30–60 days) were observed frequently in both margays. There was no evidence of gonadal seasonality in margays in either follicular or luteal activity. In ocelots, cyclic changes in estrogen excretion were observed during each month of the year; however, only one female cycled continuously. In the other two ocelots, periods of acyclicity of several months’ duration were observed. It was not possible to conclude whether tigrinas were aseasonal because estrous cyclicity was observed in only one of two individuals. In the female that cycled, a 3‐month period of acyclicity was observed in the late fall/early winter. These data demonstrate similarities among three felid species of the genus Leopardus, including evidence they are polyestrous but experience unexplained periods of ovarian inactivity. Only the margays differed by exhibiting occasional spontaneous, non‐mating–induced ovulations. Historically, these species have not bred well in captivity. However, it is hoped that understanding the biological similarities and differences among them could lead to improved management strategies that may one day result in increased reproductive success. Zoo Biol 20:103–116, 2001. © 2001 Wiley‐Liss, Inc.     

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.     

Nonsurgical embryo transfer in the scimitar-horned oryx (Oryx dammah): Birth of a live offspring

The objective of this project was to determine if modifications of methods of estrous synchronization, superovulation, embryo recovery, and transfer used successfully in other ungulates, both domestic and nondomestic, could be applied to scimitar-horned oryx (Oryx dammah). Donors were two parous females and recipients were one parous and two nulliparous females that were given a total of two cloprostenol injections at an interval of 0 and 13 or 12 days, respectively. Donors were treated with follicle-stimulating hormone (FSH-P, Schering, Kenilworth, NJ) b.i.d. for 4 days and placed with a fertile male. Seven days after the last FSH-P injection, nonsurgical uterine lavages were performed on both donors. One good-quality embryo at the morula stage was recovered and nonsurgically transferred into the right uterine horn of the parous recipient. A healthy female calf born at 247 days post-transfer represents the first known live birth of scimitarhorned oryx following embryo transfer. These results provide additional evidence that estrous synchronization and embryo transfer techniques used in other ungulates can be applied to endangered antelopes such as the scimitar-horned oryx.