Evidence from plasma progesterone concentrations for male-induced ovulation in the brush-tailed bettong, Bettongia penicillata.

Female brush-tailed bettongs, Bettongia penicillata, were housed with either an intact or vasectomized male or isolated from males in the peripartum period. Development of the quiescent corpus luteum formed at the post partum oestrus was initiated by removing the pouch young. Blood samples for analysis of plasma progesterone were collected from the females 2 days before removal of pouch young, daily for 5 or 6 days and then 2-3 times each week until 19 days after removal of pouch young. Plasma progesterone profiles were similar in pregnant and nonpregnant cycles. There was an early progesterone peak (1206 +/- 121 pg ml-1, mean +/- SEM; n = 16) between days 2 and 5 after removal of pouch young, and a second period of high concentrations (greater than 800 pg ml-1) before birth on day 17.4 +/- 0.2 (n = 16). The interval between the early peak and birth was 14 or 15 days. On five of 34 occasions, no increases in plasma progesterone concentrations occurred after removal of pouch young. On 12 of 15 occasions for 13 females that had been isolated from males post partum, plasma progesterone concentrations also remained low (less than 100 pg ml-1) and did not change after removal of pouch young. Females that showed no increases in plasma progesterone concentration after removal of pouch young had significantly lower (P less than 0.001) plasma progesterone concentrations while lactating than those females that did undergo a cycle after removal of pouch young (60 +/- 4 pg ml-1, n = 17 and 225 +/- 23 pg ml-1, n = 30, respectively). Females isolated from males post partum, and monitored until day 12 after removal of the pouch young, and that showed no increases in progesterone in this period, had ovaries that contained no corpus luteum, only corpora albicantia and numerous atretic or developing follicles. We conclude that brush-tailed bettongs are induced ovulators, a characteristic described for only one other marsupial, Monodelphis domestica, from South America.     

Comparisons of estradiol,LH and FSH patterns in pregnant and nonpregnant beagle bitches

To characterize plasma estradiol, LH and FSH patterns of secretion during the bitch estrous cycle, blood samples were obtained daily from 15 days before until 135 days after the LH surge in 10 pregnant and 10 nonpregnant beagle bitches. After an initial increase between days 15 and 10 and an expected proestrous peak, estradiol concentrations increased again from days 9-12 (corresponding to cytological metestrus) from basal values observed around day 9 after the LH surge, and remained significantly elevated throughout the luteal phase both in pregnant and nonpregnant animals. Concomitantly with the end of the luteal phase, plasma concentrations of estradiol returned to basal values in both groups. During the mid- to late-luteal phase, mean basal LH secretion was significantly elevated throughout in the pregnant relative to the nonpregnant animals. However, in nonpregnant animals, pulsatility was increased and peaks of higher amplitude were observed. The plasma FSH profiles, determined by a specific homologous RIA, differed significantly between pregnant and nonpregnant bitches during the last two-thirds of the luteal phase with a mean FSH level more elevated during pregnancy. The FSH level then decreased around parturition and low concentrations during lactation period were observed. The FSH concentrations remained steady in nonpregnant luteal phases from early luteal phase through mid-anestrus. The differences in pregnant and nonpregnant LH and FSH concentrations suggest pregnancy differences in regulation of the corpus luteum. Finally, the elevated estradiol concentrations observed during the luteal phase of both pregnant and nonpregnant animals suggest that an ovarian production of estrogens may be involved in overall corpus luteum regulation in dogs as in other species.     

Social suppression of ovarian cyclicity in captive and wild colonies of naked mole-rats, Heterocephalus glaber

To investigate the endocrine cause of reproductive suppression in nonbreeding female naked mole-rats, animals from 35 colonies were studied in captivity. Urinary and plasma progesterone concentrations were elevated in pregnant females (urine: 10.0-148.4 ng/mg Cr, 27 samples from 8 females; plasma: 3.6-30.0 ng/ml, 5 samples from 5 females; Days 21-40 of pregnancy) and cyclic breeding females (urine: 0.5-97.8 ng/mg Cr, 146 samples from 7 females; plasma: less than 1.0-35.4 ng/ml, 25 samples from 7 females). The latter group showed cyclic patterns of urinary progesterone, indicating a mean ovarian cycle length of 34.4 +/- 1.6 days (mean +/- s.e.m.) with a follicular phase of 6.0 +/- 0.6 days and a luteal phase of 27.5 +/- 1.3 days (19 cycles from 9 breeding females). In non-breeding females urinary and plasma progesterone values were undetectable (urine: less than 0.5 ng/mg Cr, 232 samples from 64 females; plasma: less than 1.0 ng/ml, 7 samples from 6 females). Breeding females had higher (P less than 0.001) plasma LH concentrations (3.0 +/- 0.2 mi.u./ml, 73 samples from 24 females) than did non-breeding females (1.6 +/- 0.1 mi.u./ml, 57 samples from 44 females). Urinary and plasma progesterone concentrations in non-breeding females from wild colonies situated near Mtito Andei, Kenya, were either below the assay sensitivity limit (urine: less than 0.5 ng/mg Cr, 11 females from 2 colonies; plasma: less than 1.0 ng/ml, 25 females from 4 colonies), or very low (plasma: 1.6 +/- 0.6 ng/ml, 15 females from 4 colonies). In captivity, non-breeding females removed from their colonies (i.e. the dominant breeding female) and either paired directly with a non-breeding male (N = 2), or removed and housed singly for 6 weeks before pairing with a non-breeding male (N = 5) may develop a perforate vagina for the first time in as little as 7 days. Urinary progesterone concentrations rose above 2.0 ng/mg Cr (indicative of a luteal phase) for the first time 8.0 +/- 1.9 days after being separated. These results suggest that ovulation is suppressed in subordinate non-breeding female naked mole-rats in captive and wild colonies, and show that plasma LH concentrations are significantly lower in these non-breeding females. This reproductive block in non-breeding females is readily reversible if the social factors suppressing reproduction are removed.     

Pregnancy-specific elevations in fecal concentrations of estradiol, testosterone and progesterone in the domestic dog (Canis familiaris)

Estradiol (E2), testosterone (T) and progesterone (P4) concentrations were determined by enzyme-immunoassay in aqueous extracts of fecal samples obtained during anestrus, proestrus, estrus and metestrus of 11 nonpregnant and 11 pregnant bitches. Fecal hormone concentrations (ng/g) changed in relation to stage of cycle. Mean fecal steroid concentrations in 22 anestrous bitches and 3 ovariectomized bitches were low and similar for E2 (53 +/- 5 and 27 +/- 2), T (60 +/- 7 and 36 +/- 6), and P4 (62 +/- 6 and 86 +/- 15). Within 0 to 3 d of the ovulatory LH surge fecal E2 reached peak concentrations (301 +/- 38). The T peaks (281 +/- 41) were coincident or 1 to 3 d later. Fecal P4 was then elevated for approximately 2 m.o. Between Days 26 and 45 after ovulation, mean fecal P4 concentrations were higher (P < 0.05) in pregnant (401 +/- 60) than in nonpregnant bitches (164 +/- 23) and peak fecal P4 concentrations in individual animals were higher (P < 0.01) in pregnant (812 +/- 121) than in nonpregnant bitches (425 +/- 97). In the same period mean concentrations of E2 (117 +/- 13 vs 61 +/- 5) and T (102 +/- 10 vs 70 +/- 6) were also higher (P < or = 0.05) in pregnant than in nonpregnant bitches. Serum E2, T and P4 concentration were positively correlated (P = 0.1) with concentration in fecal samples obtained one day after serum collection. Although serial fecal ovarian steroid concentrations demonstrate the time course of ovulatory cycles, the diagnostic value of individual fecal samples appears limited. The ratios of peak to basal values were approximately 6, 5 and 7 for E2, T and P4, respectively, and were considerably lower than ratios of 12 to 50 previously reported for serum or plasma concentrations. The results demonstrate that there are pregnancy-specific increases in P4, E2 and T production reflected in fecal concentrations. While such increases are reflected in fecal samples, they are generally not evident in serum or plasma concentrations because of increased hemodilution, metabolism and clearance in pregnant bitches. The physiological stimulus for these increases, presumably ovarian in origin, or the potential role of prolactin is not known.     

Influence of elevated ambient temperature after breeding on plasma corticoids, estradiol and progesterone in gilts

Thirty Yorkshire gilts were used to determine the influence of elevated ambient temperature during days 8 to 16 after breeding on concentrations of progesterone, corticoids and estradiol in plasma. Gilts were mated to a boar between 0800 and 1000 hr of the first day (day 0) and each subsequent day of estrus. On day 5 or 6, 22 gilts were anesthetized and a cannula was placed in the anterior vena cava. Gilts were randomly allotted to either control (23+/-1 C) or hot (35+/-1 C for 12 hr and 32+/-1 C for 12 hr daily) environmental chambers on day 8. Gilts were bled twice daily (0800 and 2000 hr) on days 9 through 16, then once daily until day 28. A third group of noncannulated gilts was bred and assigned to the control chamber. All cannulated gilts were injected intravenously with 25 IU of ACTH on day 16, and frequent blood samples were collected. Concentrations of progesterone in plasma were similar for heat-stressed and control gilts that were pregant. However, progesterone in plasma was reduced during days 13 to 19 in nonpregnant heat-stressed gilts compared to the pregnant and nonpregnant control gilts. Concentrations of estradiol in plasma were greater in nonpregnant heat-stressed gilts than in nonpregnant control and all pregnant gilts on days 10, 11 and 12 after estrus. Concentrations of corticoids and progesterone in plasma after infusion of ACTH on day 16 after breeding were reduced in heat-stressed nonpregnant gilts compared to heat-stressed pregnant, control pregnant and control nonpregnant gilts. These data indicate that reduced reproductive performance which occurs after exposure of gilts to increased ambient temperature during days 8 to 16 after breeding may be related to altered endocrine function.     

Male influence on oestrous cycles in female woolly opossum (Caluromys philander)

Plasma progesterone concentrations and the occurrence of oestrous cycles were studied in isolated woolly opossums subsequently subjected to male influences during a 40-day period. Pairing (N = 48) or exposure to male urine (N = 15) resulted in all females exhibiting oestrous during the stimulation phase, providing evidence that the activation of ovarian activity in the woolly opossum involves pheromonal cues from males. The latency of occurrence of oestrous in stimulated females depended upon their sexual state before male stimulation. In anoestrous females, the mean latency was 20.7 +/- 0.9 days (N = 35), a value which agrees with the duration of the follicular phase. In females which first entered oestrous before male stimulation, the latency of induced oestrous was inversely correlated to the date of occurrence of the previous oestrous. The inter-oestrous interval was normal (38.1 +/- 1 days, N = 5) when females were in oestrous at the beginning of male stimulation. In contrast, the inter-oestrous interval was significantly shortened (28.7 +/- 2 days, N = 7) or lengthened (51.1 +/- 1.7 days, N = 16) depending on whether females were in the luteal or follicular phases at the beginning of male stimulation. During pairing several females became pregnant and gave birth 24 +/- 0.9 days (N = 13) after copulation. In the woolly opossum, the response to male influences involves mechanisms similar to those observed in eutherians and results in enhancement and synchronization of oestrous cycles in females. Pheromonal interactions could play an important role in synchronizing oestrous cycles in wild females during the dry season, a period when animals regroup to feed on spatially localized food resources.     

Plasma concentrations of progesterone and testosterone in captive woolly opossums (Caluromys philander)

Plasma testosterone and progesterone concentrations were measured in captive woolly opossums, a didelphid marsupial originating from neotropical forests in French Guyana. Although not exposed to cyclic environmental conditions as in the field, both sexes exhibited spontaneous circannual changes in sexual hormones. Males showed synchronous variations in plasma testosterone characterized by significant elevated concentrations during April and September (8.6 +/- 1 ng/ml, N = 5) and lower levels from May to July (3.6 +/- 0.4 ng/ml). In females, synchronous periods of 2-3 successive oestrous cycles occurred. Between these periods, females remained acyclic. The oestrous cycle, determined by urogenital smears, lasted 28-45 days (n = 14) and included a 20-day spontaneous luteal phase in which progesterone concentrations reached 30-40 ng/ml plasma. Even though testosterone concentrations in paired males increased significantly in response to oestrous periods of the paired females, spontaneous circannual rhythms of sexual activity were not well synchronized between the sexes in captivity. When compared to field data, sexual activity of captive animals followed a pattern similar to that in wild animals, without any changes in males but with a delay of 3 months in females.     

Plasma concentrations of 13,14-dihydro-15-keto prostaglandin F2-alpha (PGFM), progesterone and estradiol in pregnant and nonpregnant diestrus cross-bred bitches

The canine corpus luteum (CL) typically sustains elevated plasma progesterone concentrations for 2 months or more, with a peak approximately 15-25 days after ovulation, followed by a slow decline. The processes involved in the slow, protracted regression of the CL over the remaining 1.5-2-month period in nonpregnant bitches and until shortly prepartum in pregnant bitches are not well characterized. The rapid luteolysis that occurs immediately prepartum appears to be a result of a prepartum rise in peripheral PGF. The potential role of PGF in the slow regression process in the several weeks preceding parturition and in nonpregnant bitches after 15-25 days after ovulation is not known. Therefore, plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F2-alpha (PGFM), progesterone (P4) and estradiol (E2) were determined and compared in bitches during nonpregnant diestrus (n = 9) or pregnancy (n = 8). During the gradual decrease in plasma concentrations of progesterone in both groups, the P4 pattern appeared unrelated to changes in either E2 or PGFM concentrations. The PGFM pattern was different between diestrus and pregnant bitches (P > 0.01); there was an apparent progressive but slow increase in PGFM in pregnant bitches from Days 30 to 60, followed by a large increase prior to parturition; concentrations declined immediately postpartum. However, there were no increases in PGFM during the same interval in nonpregnant bitches. Mean estradiol concentrations were sporadically elevated during the last third of pregnancy and less so in nonpregnant diestrus; there was no acute prepartum increase in estradiol associated with the PGFM increase. In summary, although there were no apparent changes in peripheral PGF2alpha concentration involved in regulating the slow protracted phase of luteal regression in nonpregnant bitches, modest increases in PGFM may play a role in ovarian function after mid-gestation in pregnant bitches. Furthermore, the acute prepartum rise in PGFM was not dependent on any concomitant increase in estradiol concentrations.     

First postpartum estrus and pregnancy in the female collared peccary (Tayassu tajacu) from the amazon

The onset of sexual cycle postpartum was described in the collared peccary (Tayassu tajacu). Serum progesterone and 17beta-estradiol profiles, vaginal smears and external genitalia were analyzed in 20 animals housed with their piglets during the first postpartum month. The appearance of external genitalia showed no variation in any of the females: a shallow, reddish vulva, and vaginal mucus were constant features throughout the study. Based on hormonal profiles and vaginal smear cell patterns, 16 (80%) of the 20 peccaries showed signs of estrus and were considered cycling. The remaining four females (20%) did no show signs of estrus confirmed by low levels of progesterone (0.9+/-0.4 ng/mL) during the first postpartum month. In the cycling peccaries, a serum 17beta-estradiol peak (53.4+/-8.1 pg/mL) was observed on Day 7+/-1 postpartum, along with a linear increase in progesterone concentration from 3 (4.3+/-2.6 ng/mL) to 11 (30.8+/-4.9 ng/mL) days after this estradiol peak. Proportions of the different cells of the vaginal epithelium also changed in these females: superficial plus intermediate cells amounted to 76% of the cell total between Days 6 and 9 postpartum, corresponding to the estradiol peak. Nine (56%) of the 16 cycling females mated, indicated by the presence of sperm cells in their vaginal smears, and 6 (67%) became pregnant, reaching term. Non-pregnant cycling females (n=10) showed a steady decrease in serum progesterone concentration from 11 to 23 days after the estradiol peak, when basal levels were attained and a new estradiol peak registered, indicating the resumption of cyclicity in these females. The time interval between the two estradiol peaks was 23.5+/-2.1 days in these females. In pregnant females, progesterone concentrations continued to rise to levels of 60 ng/mL (n=6) 23 days after mating. These findings indicate that the lactating collared peccary female can become cycling and fertile during the early postpartum period, and that a predominance of superficial plus intermediate vaginal cells can be taken as the first sign of estrus.     

Social influences on oestrous cycle length and plasma progesterone concentrations in the female lesser mouse lemur (Microcebus murinus)

Plasma progesterone concentrations were recorded during one breeding season in 19 lesser mouse lemur females living in different social conditions. The oestrous cycle length and the progesterone profile mainly depended on the social environment of the female. For totally isolated females, the oestrous cycle lasted 38 +/- 5.7 days and included a 25-30-days spontaneous luteal phase with a progesterone peak about 100 ng/ml between the 20th and 25th days after oestrus, and a prolonged preovulatory period of 10-15 days which could be considered equivalent to the follicular phase of a menstrual cycle. When females were able to communicate through olfactory, visual and auditory signals, the oestrous cycle was significantly lengthened (53.7 +/- 5.9 days). When females had tactile contacts, the oestrous cycle was further lengthened (62.7 +/- 0.8 days). This lengthening of the oestrous cycle was related to an extension of the luteal phase associated with a decrease in progesterone concentrations during this period. In females maintained with one male (paired) or with males and females (heterosexually grouped), large individual variations were shown in cycle lengths or in progesterone concentrations. In these females, cycle lengths and progesterone concentrations were inversely correlated to plasma cortisol concentrations.     

Myometrial activity and plasma progesterone and oxytocin concentrations in cycling and early-pregnant ewes

Myometrial activity and plasma progesterone (P) and oxytocin (OT) were measured in early pregnant (n = 5) and cycling (n = 5) ewes. Electromyography (EMG) leads and jugular and inferior vena cava (IVC) catheters were surgically placed in ewes about 1 wk before data collection. When ewes returned to estrus, they were bred to either an intact or vasectomized ram. Continuous EMG data were collected, and blood samples were collected twice daily from day of estrus (Day 0) until Day 18. Ewes bred with an intact ram were checked surgically for pregnancy on Day 20. Computerized, quantitative analysis of EMG events showed no difference in signal from the right to left uterine horns, and no differences between pregnant and cycling ewes (p less than 0.05) until Days 14-18 when nonpregnant ewes returned to estrus and had increased EMG activity. The mean number of EMG events 180-900 s in length decreased in pregnant ewes, but this difference was not significant (p less than 0.05). Jugular plasma progesterone (P) levels confirmed corpus luteum (CL) formation in all ewes, and no differences in P between pregnant and nonpregnant ewes were measured until Days 14-18, when cycling ewes underwent luteolysis and pregnant ewes maintained CL. IVC plasma oxytocin concentrations were increased in pregnant ewes compared to concentrations in nonpregnant ewes on Days 5-13 (p less than 0.05), and the difference was largest at Day 6 (means +/- SEM pg/ml: pregnant = 68.7 +/- 13.9, nonpregnant = 30.9 +/- 19.9).(ABSTRACT TRUNCATED AT 250 WORDS)     

Concentrations of progesterone in milk as a monitor of early pregnancy diagnosis in dairy cows

The objectives of the experiment were to evaluate the efficacy of using progesterone concentrations in milk and palpation per rectum on days 21 or 22 postbreeding to estimate pregnancy and evaluate management practices; and to investigate physiological occurrences leading to incorrect diagnosis of pregnancy when serial samples of milk were collected. Of particular interest were indications of early embyronic death and insemination of cows not in estrus. Milk samples were collected at the afternoon milking of days 0 or 1 (day 0 = day of estrus), 9 or 10, 21 or 22 and 27 or 28 following breeding in 200 lactating dairy cows. Tentative diagnosis of pregnancy was made based on concentrations of progesterone in milk on days 21 and 22 alone and on days 21 or 22 and 27 or 28. In addition all cows were palpated per rectum on days 21 or 22 postbreeding and a tentative pregnancy diagnosis was made. Pregnancy was confirmed by examination of the genital tract per rectum between 35 and 50 days after breeding. Values of 4 ng/ml or greater and/or the presence of a mature corpus luteum were considered positive signs of pregnancy. Progesterone in milk ranged from 0.1 to 18 ng/ml. On days 0 or 1, 9 or 10, 21 or 22 and 27 or 28 concentrations of progesterone in milk averaged 1.5 +/- 0.3, 11.1 +/- 0.5, 12.0 +/- 0.4 12.5 +/- 0.5 ng/ml for pregnant cows. Corresponding samples from nonpregnant cows averaged 1.2 +/- 0.2, 10.3 +/- 0.4, 3.0 +/- 0.4, 6.8 +/- 0.6 ng/ml, respectively. Ninety-six and 104 cows were classified as pregnant and nonpregnant on days 21 or 22 as compared to 78 and 118 cows diagnosed as pregnant and nonpregnant on days 21 or 22 and 27 or 28 combined. Pregnancy detection by progesterone in milk on days 21 or 22 with pregnancy determined via rectal palpation 35 to 50 days postbreeding was 77 and 100% accurate for positive and negative diagnosis, respectively. The percent agreement using progesterone in milk on days 21 or 22 and 27 or 28 combined was 95 and 100%, respectively, for positive and negative diagnosis. Diagnosis based on rectal palpation 21 or 22 days postbreeding was 63 92 (69%) and 76 88 (87%) for pregnant and nonpregnant cows, respectively. Ten of the 200 cows had progesterone concentratins in milk of > 4 ng/ml at the time of breeding. Six of these cows were pregnant from a previous insemination. The other four cows were nonpregnant and were inseminated during the luteal phase of the cycle. In conclusion, measurement of progesterone in milk is a useful tool in early detection of pregnant and nonpregnant cows and may be useful in detecting reproductive problems in a dairy herd. It will probably be most useful when used in combination with later pregnancy diagnosis per rectum .     

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

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

Effect of testosterone and bovine follicular fluid on concentrations of luteinizing hormone and follicle-stimulating hormone in plasma of castrated rams that are homozygous carriers or non-carriers of the Booroola fecundity gene.

Castrated adult FecBFecB and Fec+Fec+ Booroola rams were injected with charcoal-treated bovine follicular fluid (bFF) (a source of inhibin-like activity) or given testosterone implants to examine whether the fecundity gene (FecB) influences sensitivity to negative feedback hormones in males. Mean concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) did not differ between genotypes before treatment. In Expt 1, injections of 5 ml bFF, but not of 1 ml (each given four times at intervals of 8 h), significantly (P < 0.05) depressed concentrations of LH and FSH, but there was no effect of genotype. After treatment, gonadotrophin concentrations returned to pretreatment values and for 2-2.5 days scaled (divided by pretreatment mean) LH values (235 +/- 49 for FecBFecB and 96 +/- 26% for Fec+Fec+ rams; P < 0.05) and scaled FSH values (106 +/- 5 for FecBFecB and 85 +/- 5% for Fec+Fec+ rams; P < 0.05) were significantly higher in FecBFecB than in Fec+Fec+ rams in the group that received 5 ml bFF. Irrespective of genotype, treatment with 5 ml bFF did not reduce mean FSH to concentrations observed in testis-intact rams. In Expt 2, Silastic envelopes were implanted subdermally to give physiological or supraphysiological circulating concentrations of testosterone. Both doses significantly reduced scaled LH values in a biphasic manner, such that there was an initial suppression followed by a short-lived increase. During the initial period of suppression in the lower dose group, mean scaled LH values were significantly higher in FecBFecB than in Fec+Fec+ rams (48.3 +/- 7.5 versus 23.1 +/- 5.5%; P < 0.05). Low doses of testosterone decreased LH pulse frequency in both genotypes but decreased (P < 0.05) pulse amplitude and mean concentrations in the Fec+Fec+ animals only. In nonimplanted control rams, mean LH concentrations (in samples taken every 10 min for 12 h) were significantly lower in FecBFecB than in Fec+Fec+ rams (0.6 +/- 0.2 versus 1.3 +/- 0.1 ng ml-1; P < 0.05). The mean FSH response to testosterone was not related to genotype. These data suggest that expression of the FecB gene results in an altered sensitivity of the pituitary gland to changes in negative feedback from testicular hormones and that, irrespective of genotype, neither testosterone nor inhibin-like activity alone can fully control FSH secretion in castrated rams.     

Serum androstenedione and testosterone concentrations during pregnancy and nonpregnant cycles in dogs

Concentrations of testosterone and of androstenedione were determined by radioimmunoassay in serum samples collected every 2-5 days throughout the periovulatory and luteal phases of the ovarian cycles of pregnant and nonpregnant beagle bitches. Testosterone levels were consistently lower than those of androstenedione, reached peaks of 29 +/- 4 ng/dl near the time of the preovulatory luteinizing hormone peak, and were reduced to near the limits of detection (less than or equal to 5-10 ng/dl) throughout the luteal phase. Androstenedione levels reached preovulatory peaks of 73 +/- 13 ng/dl, were 54 +/- 7 ng/ml during early estrus, increased (P less than 0.05) to early luteal phase peaks of 76 +/- 8 ng/dl between Days 6 and 18, and then declined to 41 +/- 5 ng/dl by Day 35-40 in both pregnant (n = 8) and nonpregnant (n = 4) bitches. Subsequent protracted increases in androstenedione occurred in 4 of 8 pregnancies but in none of the nonpregnant bitches. From Days 42 to 64 the differences in mean levels between pregnant (45 +/- 2 ng/ml) and nonpregnant (32 +/- 3 ng/ml) bitches was not significant (P greater than 0.05). At parturition androstenedione levels fell (P less than 0.05) abruptly from 39 +/- 7 to 13 +/- 3 ng/dl. These results suggest that, in the bitch, androstenedione is the major circulating androgen during the follicular and luteal phases and that patterns of androstenedione levels during the luteal phase parallel those reported for progesterone in pregnant and nonpregnant bitches, including maintenance of elevated levels throughout gestation and an abrupt decline at parturition.     

Circulating progesterone and oestradiol-17 beta concentrations in cyclic Cape porcupines, Hystrix africaeaustralis

The general pattern of steroid secretion during the 30-day oestrous cycle of the Cape porcupine is that of a surge (25-176 pg/ml) in oestradiol-17 beta secretion at the time of perforation of the vaginal closure membrane, followed by an increase in progesterone concentrations, the latter attaining peak values (mean 5.9 +/- 2.1 ng/ml) 8-19 days (13.8 +/- 2.8 days) after vaginal opening. Copulation occurred after the oestradiol-17 beta surge and the length of the luteal phase of the cycle varied from 21 to 35 days (29.3 +/- 4.7 days), this representing 93% of the length of the cycle. Perforation of the vaginal closure membrane was not always accompanied by an increase in oestradiol-17 beta levels and some instances (19%) of vaginal opening were not followed by an increase in progesterone secretion. The hormonal characteristics of the oestrous cycle of females housed with vasectomized males were similar to those of females housed with intact males.     

PLASMA PROGESTERONE CONCENTRATIONS MEASURED USING AN ENZYME-LINKED IMMUNOSORBENT ASSAY USEFUL FOR DIAGNOSING PREGNANCY IN HARBOR SEALS (PHOCA VITULINA)

Wild-caught female harbor seals ( Phoca vitulina ) were classified as sexually mature or immature on the basis of standard body length (< 125 cm immature, > 125 cm mature) and plasma progesterone concentrations measured using an enzyme-linked immunosorbent assay (ELISA), a technique usable in the field. Sexually mature females were classified as pregnant or non-pregnant on the basis of their plasma progesterone concentrations. Of 28 wild mature female harbor seals caught in the Moray Firth, N.E. Scotland, between the end of February and the end of May, 79% had plasma progesterone concentrations greater than 60 nmol liter⁻¹, the lowest plasma progesterone concentration measured in one of eight females later observed with a pup, and were diagnosed as pregnant. A linear discriminant function, calculated to provide a method of distinguishing pregnant and non-pregnant females, predicted 100% of non-pregnant females and 95.8% of pregnant females using plasma progesterone concentration, standard length, and month of capture as parameters. Plasma progesterone concentrations were less than 30 nmol liter⁻¹ in all mature and immature males and immature females. In mature females plasma progesterone concentrations ranged from 0-318 nmol liter⁻¹.     

Reproductive endocrine patterns in captive female and male red wolves (Canis rufus) assessed by fecal and serum hormone analysis

Reproductive steroid profiles in female (n=13) and male (n=5) red wolves (Canis rufus) were characterized in fecal samples collected during the breeding season (December—May) and over a 1 year period, respectively. Blood samples from females (n=12) also were collected during the periovulatory period for luteinizing hormone (LH) and steroid analysis. High performance liquid chromatography (HPLC) of fecal extracts determined that estradiol and estrone constituted the major and minor forms, respectively, of fecal estrogen metabolites. Although native progesterone was present, pregnane metabolites predominated as the major forms of fecal progestins. HPLC analysis of fecal extracts from males revealed no native testosterone, but rather the predominance of more polar androgen metabolites. Based on hormone profiles and/or pup production, females were classified as pregnant (n=3), ovulatory‐nonpregnant (n=9), or acyclic (n=3). Longitudinal monitoring of females indicated no pregnancy‐specific differences in concentrations of either fecal progestagen or estrogen metabolites compared to ovulatory‐nonpregnant individuals; however, baseline progestagen concentrations were consistently elevated in acyclic females. There was good correspondence between serum and fecal steroid concentration during the periovulatory period. A rise in serum estrogens preceded the ovulatory LH surge which was then followed by a significant progesterone rise during the luteal phase. In males, changes in fecal androgen metabolite concentrations coincided with photoperiod fluctuations, increasing in late autumn and reaching peak concentrations during mid‐ to late winter just before the start of the breeding season. Collectively, these results serve as a database of ovarian and testicular endocrine events in this species, which can be utilized in population management and application of assisted reproductive technologies. Zoo Biol 21:321–335, 2002. © 2002 Wiley‐Liss, Inc.     

Validation of a serum immunoassay to measure progesterone and diagnose pregnancy in the West Indian manatee (Trichechus manatus)

The objective was to validate a high-sensitivity chemiluminescent assay of serum progesterone concentrations for pregnancy diagnosis in manatees. Assay analytical sensitivity was 0.1 ng/mL, with mean intra- and inter-assay coefficients of variation of 9.7 and 9.2%, respectively, and accuracy had a mean adjusted R(2) of 0.98. Methods comparison (relative to Siemen's Coat-A-Count RIA) demonstrated r=0.98, Deming regression slope of 0.95, and an intercept of 0.01. Based on ROC analysis, a progesterone concentration >or=0.4 ng/mL was indicative of pregnancy. Assay results were not significantly altered by two freeze-thaw cycles of samples. Characteristic progesterone concentrations during pregnancy were Months 1-4 (1.7-4.7 ng/mL), 5-8 ( approximately 1.0 ng/mL), and 10 and 11 (0.3-0.5 ng/mL), whereas two late-pregnant females with impending abortion had progesterone concentrations of 0.1 ng/mL. Among pregnant females, maximum progesterone concentrations occurred in autumn (3.9+/-1.8 ng/mL), and were greater during all seasons than concentrations in non-pregnant females (0.1-0.2 ng/mL). Progesterone concentrations were also significantly higher in pregnant females than in non-pregnant females and males. This highly sensitive, specific, and diagnostic assay will be valuable for monitoring pregnancy and abortion in manatees.     

Characteristics of the oestrous cycle of Iberian red deer (Cervus elaphus hispanicus) assessed by progesterone profiles

This study examines the length of the oestrous cycle in 16 Iberian red deer females assessed by means of changes in progesterone concentrations, along with the changes in the profile of this hormone. Samples were collected three occasions per week from the week after calving (15 May to 15 June) up to May of the following year. The oestrous cycle lasted 19.57+/-0.29 days (range 10-27 d) calculated in 130 oestrous cycles examined. Progesterone titres did not rise above 0.5 ng/ml in the follicular phase, except in four samples. The maximum peak in progesterone concentration during the luteal phase remained above 1 ng/ml in most cases. Twenty-five percent of the individuals studied (4 out of 16) showed an oestrous cycle lasting shorter than the mean (15.2+/-0.30 days) before the start of the reproductive season, followed by a period of sexual inactivity. The standard progesterone profile in natural oestrous cycles rose from basal levels to those above 0.5 ng/ml four days after onset of oestrus, reached a peak of 1.71+/-0.07 ng/ml and then declined to less than 0.2 ng/ml after day 20. Following the rapid decline of progesterone after day 14, the concentration remained around the baseline level of 0.1 to 0.2 ng/ml during the immediate pre- and post-ovulatory phase of the cycle.