John D. Harder

Male pheromone stimulates ovarian follicular development and body growth in juvenile female opossums (Monodelphis domestica)

Female opossums are induced into estrus by direct exposure to a non-volatile pheromone in male scentmarks. Juvenile females develop this responsiveness by 150 days of age (days), and earlier (130 days) if exposed to male pheromone beginning at 90 days. The goal of this study was to examine the effect of male pheromone on body growth and ovarian follicular development in young opossums. Females (n = 28) were weaned at 56 days and caged individually with ad libitum food and water from 84 days. Body mass was recorded 2–3 times weekly and the onset of estrus was monitored by urogenital sinus cytology. Exposed females were given continuous access to adult male scentmarks from 90 days and randomly selected for necropsy at 105 and 130 days. Unexposed females were isolated from direct contact with males and their scentmarks and randomly selected for necropsy at 90, 105, 130, and 150 days. Exposed females were larger (63.5 ± 1.1 g) than unexposed females (56.6 ± 1.1 g) at 130 days, and 4 of 5 had expressed estrus or proestrus. Uterine mass at 130 days was higher (P < 0.05) in exposed (129.8 ± 28.8 mg) than in unexposed (25.4 ± 6.7 mg) females, none of which expressed estrus by 150 days. The mean number of developing, antral follicles per female increased from 1.5 ± 1.5 at 90 days to 17.8 ± 5.2 at 130 days. Mean diameter of developing antral follicles at 130 days was larger in exposed (534 ± 54 μm) than in unexposed females (393 ± 4 μm). The results of this study demonstrate that pheromonal induction of first estrus in juvenile opossums is associated with an increased rate of body growth and follicular development.

Fecal corticosterone concentrations and reproductive success in captive female southern white rhinoceros

Prolonged or frequent secretion of adrenal glucocorticoids in response to aversive stimuli can negatively impact reproduction. Because female southern white rhinoceros (Ceratotherium simum simum) reproduce poorly in captivity, we compared fecal corticosterone metabolite concentrations among parous, nulliparous, and adolescent females and examined social and physical aspects of the captive environment that might be related to differences in corticosterone metabolite concentrations. Aggression, dominance, sexual and play interactions, social group size and composition, enclosure size, and other housing characteristics were assessed though behavioral observations and review of historical and institution records. Concentrations of metabolized corticosterone in fecal samples were analyzed by enzyme immunoassay. The proportion of nulliparous females did not differ (p>0.05) between subordinate and dominant animals, and subordinates did not have a higher mean fecal corticosterone concentration than dominants (p>0.05). Of the behaviors examined, only the frequency of sexual play behaviors differed (p<0.05) between dominants and subordinates. Average corticosterone concentrations differed (p<0.05) across housing institutions but were not consistently elevated (p>0.05) for females housed in most of the environmental conditions assessed. Housing with a female companion known from adolescence, however, tended to be associated (p=0.06) with a lower mean corticosterone concentration than that when housing with a female companion introduced during adulthood or no female companion. Wild-caught females had a higher (p<0.05) average corticosterone concentration than captive-born females. Average corticosterone concentration did not differ (p>0.05) between acyclic and cycling, or nulliparous and parous females.

Evidence for Spontaneous Postlactational Estrus in Gray Short-Tailed Opossums (Monodelphis domestica)

Abstract Previous studies of the gray short-tailed opossum have shown that ovarian activity and estrus are induced by male pheromones, but we recently documented urogenital sinus (UGS) estrus in postlactational females despite their isolation from the male stimuli known to be associated with induced estrus. Body weights and UGS smears were collected after removal of pups in midlactation (19–37 days postpartum), after weaning (55–61 days postpartum), or after pheromone exposure. Estradiol was measured by RIA in plasma samples collected from dams during lactation, after separation from pups, and at estrus. Average days to UGS estrus from pup removal or initial pheromone exposure differed (P < 0.05) only between the midlactation and pheromone exposure groups. Postlactational females showed a decrease in body weight from the time of pup removal or weaning to estrus, which contrasts with the increase seen in pheromonally stimulated females. Plasma estradiol was elevated at estrus in all groups, and females that were paired with males at postlactational estrus mated and produced litters. This study demonstrates that gray short-tailed opossums consistently experience estrus within 2 wk of weaning their young and that postlactational estrus appears to be hormonally and behaviorally equivalent to estrus induced by direct exposure to male pheromones.

Urinary progesterone in free‐ranging red howler monkeys (Alouatta seniculus): Preliminary observations of the estrous cycle and gestation

The goals of this study were to develop and validate a radioimmunoassay (RIA) for measurement of unconjugated progesterone (P) concentrations in the urine of red howler monkeys (Alouatta seniculus) and to use urinary P profiles to characterize the reproductive cycle of this species. Analysis of P profiles from two females provided a preliminary estimate of the length of the estrous cycle (mean days ± S.E.M. = 29.5 ± 1.5; n = 2), and indicated that one female red howler copulated throughout two apparent estrous cycles. Urinary P concentrations during two confirmed pregnancies (211.8 ± 29.7 ng P/ml) were higher (P < 0.05) than during the luteal phase (77.4 ± 10.6 ng P/ml; n = 4) of the cycle. Am. J. Primatol. 51:257–263, 2000.

Reproductive energetics of the virginia opossum compared with some eutherians

Abstract 1. 1. Metabolic rates (MR) of 8 lactating opossums and their litters were measured by respirometry. 2. 2. MR was 92% higher throughout lactation when compared to nonreproductive adults. 3. 3. Total resting metabolic cost for an opossum nursing throughout pouch life was estimated at 400 kcal. 4. 4. Comparison of total energetic costs or of efficiencis during gestation and lactation in several mammals does not support assumptions of the inferiority of the marsupial reproductive strategy.

Ecology of Marsupials in Two Amazonian Rain Forests in Northeastern Peru

Species diversity, relative abundance, and seasonal variability in reproductive activity of marsupials were examined at two locations in tropical rain forests in northeastern (Amazonian) Peru during wet and dry seasons. Eight species of opossums (Didelphimorphia) were captured in a single 11-ha plot, but only four ( Didelphis marsupialis, Marmosops noctivagus Philander opossum , and Philander andersoni ) were common. Large differences with respect to species diversity and abundance of opossums were found between two similar rain-forest sites. Reproductive activity in opossums was positively related to rainfall and fruit production.

Characterization of the estrous cycle and assessment of reproductive status in Matschie’s tree kangaroo (Dendrolagus matschiei) with fecal progestin profiles

The population of Matschies tree kangaroos (Dendrolagus matschiei) held in North American zoos has declined to critically low numbers, and information on the reproductive biology of tree kangaroos is limited. The objectives of this study were to (1) characterize the temporal features of the estrous cycle through the measurement of fecal progesterone metabolite (i.e., progestin) concentrations and (2) determine the reproductive status of female tree kangaroos in the captive population of North America through the identification of estrous cyclicity. Fecal pellets and observations of estrous behaviors were collected from 16 captive female tree kangaroos. Fecal pellets were sampled and extracted with methanol, and progestin concentrations were quantified using a radioimmunoassay (RIA) for progesterone and its metabolites. A progestin profile was obtained for each female by plotting fecal progestin concentrations for every third day over a 120-day period. Profiles for 12 of 16 females showed evidence of estrous cyclicity (P<0.01). The mean length of the estrous cycle was estimated at 58.9+/-2.4 days (n=11). Progestin concentrations were low during the first 15-20 days of the luteal phase and remained elevated above baseline only during the last 30.2+/-3.2 days of the luteal phase, which averaged 46.6+/-2.5 days in duration. The progestin profile observed in the estrous cycle of Matschies tree kangaroos in this study is very similar to that seen in the non-pregnant cycle of several other species in the family Macropodidae.

Estrogen in peripheral plasma during postnatal development in gray short-tailed opossums

Plasma samples obtained from gray short-tailed opossums (Monodelphis domestica) at selected ages through adulthood were assayed for estrogen (E). Levels of E in one mixed-sex plasma pool of animals aged postnatal day (pd) 4 and one of two mixed-sex plasma pools of animals aged pd 8 were over 300 pg/ml. On pd 16, E levels in males and females averaged 30 and 47 pg/ml, respectively. While no significant sex differences in E levels were seen on pd 30 or pd 60, mean E levels for animals on pd 30 were significantly higher (275 pg/ml in males and 181 pg/ml in females) than on pd 60 (78 pg/ml in males and 85 pg/ml in females) or pd 145 (adults). In adult animals, estrogen levels in females averaged 54 pg/ml; all adult male E levels were below the limit of sensitivity of the assay. Maternal E levels, which did not vary significantly by age of litter, averaged 10 pg/ml overall. These findings are discussed with respect to possible significance of high E levels in developing marsupials for sexual differentiation and general brain development.

Effect of pregnancy on uterine constituents of the virginia opossum

1. 1. Maternal reproductive physiology during opossum gestation was examined by comparison of the biochemical composition of uteri (N = 21) from days 3, 7 and 11 of the luteal phase of the oestrous cycle and of pregnancy. 2. 2. No differences occurred in total protein, glucose, and percent water. 3. 3. Glycogen was similar in both reproductive states on days 3 and 11, but pregnant uteri had significantly lower (P < 0.05) glycogen levels on day 7 than non-pregnant uteri (range in mg/g dry weight: 4.4–7.8 and 8.5–15.8, respectively). 4. 4. Mobilization of uterine glycogen might be induced by biophysical stimuli from expanding blastocysts that generate myometrial contractions, thereby distributing the embryos throughout the uterine lumen.

Progesterone receptor in the forebrain of female gray short-tailed opossums: Effects of exposure to male stimuli

Progesterone receptor immunoreactivity (PRir) in brain areas involved in reproductive behavior in eutherian species was examined for the first time in a female marsupial, the gray short-tailed opossum (Monodelphis domestica, hereinafter, opossum). PRir in nuclei of neurons, measured as area covered by stained nuclei, was seen in the arcuate nucleus (Arc); anteroventral periventricular nucleus (AVPv); bed nucleus of the stria terminalis (BST); medial preoptic area (MPOA), and ventromedial hypothalamus (VMH), but not in control areas adjacent to the hypothalamus or cortex. Female opossums are induced into cytological, urogenital sinus (UGS), estrus by male pheromones and into behavioral estrus, i.e., receptivity, by pairing with a male, and both estradiol (E) and progesterone (P) are involved in induction of receptivity in intact and ovariectomized females. PRir in the AVPv, MPOA, and VMH was very low in females that had never been exposed to males or their scent marks, i.e., naïve anestrous (NVA) females, and either previous or current exposure to males or their scent marks was associated with elevated PRir. PRir was significantly higher in the AVPv and MPOA of anestrous females with previous but no current exposure to males and their scent marks, i.e., experienced anestrous (EXPA) females, than in NVA females, but PRir was significantly lower in the MPOA and VMH of EXPA females than in females that were behaviorally receptive and had recently copulated, i.e., behavioral receptive estrous (BRE) females. PRir was higher in the VMH of both UGS estrous (UGSE) and BRE females compared to that in EXPA animals, but PRir did not differ between UGSE and BRE females in any of the 3 brain areas examined, including the MPOA These results provide evidence that pheromonal induction of estrus and sexual receptivity in opossums is associated with elevation of PRir in the VMH and MPOA and that prior exposure to males or their pheromones, even in the absence of current male stimuli, is associated with persistent elevation of PRir in the AVPv and MPOA.