Ovarian hormone concentrations and genital swelling patterns in female chimpanzees with Norplant implants

The purpose of this study was to assess the effects of Norplant implants on ovarian hormone secretion and genital swelling patterns in female chimpanzees. Urine samples were collected during one natural cycle before (n = 5 females) and one cycle ∼6 months following (n = 7 females) insertion of Norplant implants. Genital swelling data were recorded daily for all seven females. During Norplant treatment, two females exhibited minimal ovarian activity consistent with anovulation. Hormone data from five subjects indicated both follicular and luteal activity, suggesting that ovulation may have occurred during Norplant treatment. Among cycles presumed to be ovulatory, the concentration of estrone glucuronide did not differ from concentrations during the control phase. However, mean (F = 12.253, P = 0.001) and peak (F = 4.127, P = 0.003) concentrations of pregnanediol glucuronide were significantly lower during Norplant treatment. Cycle duration did not differ during Norplant treatment (F = 1.415, P = 0.200); however, duration of the total swelling phase (F = 3.311, P = 0.0011) and full swelling phase was shorter (F = 5.906, P = 0.011), whereas number of days with partial swelling increased (F = 2.635, P = 0.011). The results of this study indicate that Norplant provided effective contraception while not inhibiting ovarian hormone secretion or the expression of cyclic genital swelling in the majority of the chimpanzee subjects. Zoo Biol 16:209–223, 1997. © 1997 Wiley-Liss, Inc.     

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.     

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.     

Effect of Feeding Inorganic Chromium on Growth Performance, Endocrine Variables, and Energy Metabolites in Winter-Exposed Buffalo Calves (Bubalus bubalis)

We investigated the effect of chromium (Cr) supplementation on the growth performance, energy metabolites, and hormonal variation in winter-exposed buffalo calves. Twenty-four female buffalo calves were randomly allotted to four dietary treatments (n?=?6) for a period of 120 days. Feeding regimen was the same in all the groups, except the animals in the four respective groups were additionally supplemented with 0.0, 0.5, 1.0, and 1.5 mg of Cr/kg DM in the form of CrCl₃.6H₂O. Calves were monitored daily for physiological variables and dry matter intake (DMI). Blood samples were collected at fortnightly intervals from each buffalo calves to measure concentrations of hormones (insulin, cortisol, and growth hormone), energy metabolites (glucose and non-esterified fatty acids), and plasma mineral levels. After 120 days of feeding trial, buffalo calves fed with Cr had lower (P?<?0.05) circulating plasma concentrations of glucose, insulin, and cortisol hormones, whereas plasma thyroid hormone and non-esterified fatty acids concentrations were found similar (P?>?0.05) among all the treatments. The results suggested that dietary Cr supplementation influenced plasma Cr levels without affecting the plasma concentrations of other trace minerals. However, physiological variables, nutrient intake, and growth performance of buffalo calves did not differ among all treatments (P?>?005). In summary, the current study showed that supplementation of Cr at the level of 1.0 and 1.5 mg of Cr/kg DMI was more effective in improving glucose utilization by increasing potency of insulin hormone and reducing concentration of cortisol hormone. Results also suggested that supplemental Cr also improves blood plasma Cr levels.     

Effects of estradiol on renal cyclic guanosine monophosphate and oxidative stress in spontaneously hypertensive rats

Background: Evidence suggests that estradiol offers protection against the development of cardiovascular and renal pathologies, although the mechanisms involved are still under investigation. The nitric oxide (NO) pathway regulates blood pressure and kidney function, and estradiol is associated with increases in NO bioavailability. We hypothesized that in female spontaneously hypertensive rats (SHRs), estra-diol increases NO bioavailability, activates the NO synthase (NOS) pathway, and suppresses superoxide production compared with rats that underwent ovariectomy (OVX).Objective: The goal of this study was to determine whether estradiol regulates the NO/cyclic guanosine monophosphate (cGMP) pathway and superoxide levels in the kidneys of female SHR.Methods: Three types of SHRs were studied: gonad-intact females, OVX rats, and OVX rats with estra-diol replacement (OVX+E). Renal cortical cGMP levels were measured to assess NO bioavailability. NOS enzymatic activity, NOS protein expression, basal superoxide production, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity were measured in the renal cortex.Results: Fifty-six SHRs were included in the study (17 intact females, 21 OVX rats, 18 OVX+E rats). Mean (SEM) cGMP levels were significantly lower in the renal cortex of OVX rats (0.03 [0.008] pmol/mg, n = 5) than in intact females (0.1 [0.02] pmol/mg, n = 6; P < 0.05), and estradiol restored cGMP levels to those seen in intact females (0.1 [0.01] pmol/mg, n = 5; P < 0.05). Despite a decrease in cGMP following OVX, renal cortical NOS activity, NOS1 and NOS3 protein expression, and the phosphorylation status of NOS3 were comparable among the 3 groups (n = 7–9 per group). However, mean basal superoxide production in the renal cortex was higher in OVX rats (3.2 [0.3] cpm/mg, n = 12) than in intact females (1.9 [0.3] cpm/mg, n = 8; P < 0.05) and lower in OVX+E rats (1.3 [0.3] cpm/mg, n = 9; P < 0.05). Mean NADPH oxidase activity was comparable in the renal cortex of intact females and OVX rats (81 [4] and 83 [12] cpm/35 μg, respectively [n = 5 per group]). OVX+E rats had significantly lower mean renal cortical NADPH oxidase activity than did rats in the other groups (45 [6] cpm/35 μg, n = 6; P < 0.05), and the decrease in activity was accompanied by a decrease in p22^phox protein expression.Conclusions: In vivo manipulations of estradiol levels influenced renal cortical NO bioavailability, as assessed indirectly by cGMP measurements. The decrease in cGMP following OVX was not due to alterations in the activity or expression of NOS.     

Characterization of reproductive cycles and adrenal activity in the black‐footed ferret (Mustela nigripes) by fecal hormone analysis

The reproductive cycle of the black‐footed ferret (Mustela nigripes) was characterized by enzyme immunoassay (EIA) analysis of ovarian fecal steroids (estradiol, progestins) in 29 females over two consecutive breeding seasons. Estrous status was determined by measuring the vulva size and examining the percentage of superficial cells in vaginal lavages. Mean fecal estradiol concentrations were correlated with vulval area (r = 0.370, P < 0.0001) and the percentage of superficial cells (r = 0.380, P < 0.0001). Ovulation resulted in a rise in fecal progestin concentrations 5 days after breeding that differed (P < 0.05) between pregnant (n = 14) and pseudopregnant (n = 12) females during the late luteal phase (days 12–40), with concentrations remaining higher in pregnant animals. Gestation length was 41.3 ± 0.7 days with 3.6 ± 0.4 kits produced per female. Litter size correlated significantly (P < 0.05) with fecal estradiol, but not progestins during the 12 to 40 days after breeding. Females failing to breed (n = 3) remained in estrus for 31 ± 6.2 days before ovulation induction with human chorionic gonadotropin. Adrenal activity in male (n = 4) and female (n = 6) black‐footed ferrets was monitored by quantifying fecal corticoid metabolites after a series of manipulations (physical restraint, intramuscular saline, intramuscular gel adrenocorticotropic hormone (ACTH), intramuscular liquid ACTH). A significant (P < 0.0001) increase in fecal corticoids above the pre‐treatment baseline occurred 20 to 44 hours after restraint (five of 10 animals), saline (six of nine), gel ACTH (seven of 10), and liquid ACTH (nine of 10) treatments. Immunoreactivity of high‐performance liquid chromatography–separated fecal elutes was compared using antibodies against cortisol and corticosterone. The cortisol EIA demonstrated immunoreactivity that co‐eluted with ³H‐cortisol, whereas a corticosterone radioimmunoassay detected a metabolite peak that co‐eluted with ³H‐corticosterone in addition to a slightly less polar and one considerably more polar peak. Despite recognizing different metabolites, both assays produced similar temporal profiles of corticoid excretion after manipulation. This study provides new information on the black‐footed ferret regarding differences in fecal steroid excretion patterns between pregnancy and pseudopregnancy and the potential application of fecal corticoid metabolite monitoring for evaluating responses to stressors associated with practices used in breeding management. Zoo Biol 20:517–536, 2001. © 2002 Wiley‐Liss, Inc.     

The Potential of Cr3 [Triaqua‐μ3‐Oxo‐Hexa‐μ‐Propionatotrichromium(III) Chloride] to Reduce Birth Defects in the Offspring of Diabetic CD‐1 Mice

Diabetes mellitus is a growing concern worldwide and leads to multiple complications during pregnancy. Pharmacologic doses of chromium (Cr) have been linked with improving insulin sensitivity and other positive benefits in the treatment of diabetes in animal models. By using streptozotocin induced hyperglycemia in female CD‐1 mice, reproductive outcomes of diabetic and chromium‐dosed diabetic females were examined. After dosing 10 mg/kg Cr in the form of triaqua‐μ₃‐oxo‐hexa‐μ‐propionatotrichromium(III) chloride or Cr3 during gestation days 8–16 (GD8–GD16), all females were sacrificed on gestation day 17 (GD17) and examined for maternal weight gain. The fetuses were examined for gross malformations and for skeletal malformations. The offspring of Cr3‐dosed females tended to have a reduction in the incidence of supernumerary ribs. While hyperglycemia still had negative impacts on the health of dams and their offspring, administration of Cr led to an apparent trend in the reduction in the number of malformations and incidence of supernumerary ribs compared to those of untreated diabetic mothers     

Describing Ovarian Cycles,Pregnancy Characteristics,and the Use of Contraception in Female White‐Faced Marmosets,Callithrix geoffroyi

Endocrine data and characteristics of nonconceptive ovarian cycling and pregnancy are limited within the genus Callithrix to the common marmoset (C. jacchus) and Wied's black tufted‐ear marmoset (C. kuhlii). This article presents patterns of urinary pregnanediol‐3‐glucuronide (PdG) excretion, as determined by enzyme immunoassay, throughout the course of ovarian cycling and pregnancy in white‐faced marmosets (C. geoffroyi). Furthermore, characteristics of reproductive parameters including litter size, duration of gestation, maternal age, and information about ovarian cycling following administration of contraceptives are also described. A steep increase in PdG, an indication of ovulation, characterizes normative ovarian cycles, with peak‐to‐peak intervals between cycles being 27.82 ± 1.49 days in length. PdG excretion (μg/mg Cr) across pregnancy peaked during the 1st and 2nd trimesters (1st = 20.71 ± 2.98, 2nd = 21.16 ± 2.60) and declined gradually to near preconception levels over the 3rd trimester until parturition (3rd = 5.74 ± 1.60). Gestation lasted 148.55 ± 1.89 days. Most pregnancies (82.8%) resulted in an immediate postpartum ovulation (PPO) of 17.45 ± 2.22 days with 58.3% of PPOs resulting in conception. No differences in PdG excretion during the 1st trimester between full pregnancies and miscarriages were found, and pregnancy characteristics such as litter size, duration of gestation, and maternal age were not associated with PdG concentrations. Administration of cloprostenol resulted in shorter peak‐to‐peak cycle durations, but ovulation was detectable with similar concentrations of peak PdG to a normal nonconceptive cycle. Conversely, medroxyprogesterone acetate (DMPA) injections resulted in little to no PdG excretion across the ovarian cycle. Both methods of contraception providing effective prevention of conception. Overall, these results show that strong similarities in reproductive parameters persist within the genus Callithrix and to a lesser extent across the Callitrichidae family. Am. J. Primatol. 74:1044‐1053, 2012. © 2012 Wiley Periodicals, Inc.     

Induction of ovarian activity and ovulation in an induced ovulator,the maned wolf (Chrysocyon brachyurus), using GnRH agonist and recombinant LH

Assisted reproductive techniques, such as ovarian manipulation and artificial insemination, are useful for enhancing genetic management of threatened wildlife maintained ex situ. In this study, we used noninvasive fecal hormone monitoring to investigate (1) the influence of pairing with a male on endocrine responses of female maned wolves (Chrysocyon brachyurus) to a GnRH agonist (deslorelin) and (2) the efficiency of recombinant LH (reLH) on ovulation induction in females housed alone. Deslorelin (2.1 mg Ovuplant) was given to females that were either paired with a male (n = 4) or housed alone (n = 7); the implant was removed 7 to 11 days postimplantation. Three of seven singleton females were injected with reLH (0.0375 mg) on the day of implant removal, whereas the remaining females (n = 4) did not receive the additional treatment. Fecal samples were collected 5 to 7 days/wk from all females starting 11 days prior to hormone insertion until at least 70 days post implant removal for a total of 11 hormone treatment cycles. Fecal estrogen and progestagen metabolites were extracted and analyzed by enzyme immunoassay. Evidence of ovulation, demonstrated by a surge of estrogen followed by a significant rise in progestagen, occurred in all paired females. Three of the four singleton females that did not receive reLH treatment exhibited no rise in progestagen after an estrogen surge. All singleton females treated with reLH exhibited a rise in fecal progestagen after injection, indicating ovulation. In conclusion, deslorelin is effective at inducing ovarian activity and ovulation in paired female maned wolves; however, exogenous reLH is needed to induce ovulation in females housed alone. The findings obtained from this study serve as a foundation for future application of artificial insemination to enhance genetic management of this threatened species ex situ.     

Combined Olfactory Contact with the Parent Colony and Direct Contact with Nonbreeding Animals Does Not Maintain Suppression of Ovulation in Female Naked Mole-Rats (Heterocephalus glaber)

The study investigated the role of odor cues from two naked mole-rat colonies, in conjunction with behavioral cues from nonbreeding colony members, in maintaining suppression of ovulation in subordinate female naked mole-rats isolated from the two parent colonies. Four high ranking nonbreeding female naked mole-rats were removed from their respective parent colonies and singly housed in separate burrow systems. For a 64-day period, the removed females were maintained in daily odor contact with their parent colony by daily rotating soiled bedding material between the parent colony and the burrow systems of removed females. In addition, subsets of nonbreeding animals from the respective parent colony were regularly moved into the burrow systems of removed females for 2-day periods during this 64-day period. Removed females were therefore in continual social contact with subsets of parent colony animals except for the breeding pair. All four removed females exhibited raised levels of urinary progesterone (< 2 ng/mg Cr) indicative of the onset of ovarian function within 3 days of being separated from the parent colony. Removed females exhibited a normal ovulatory cycle with levels of progesterone remaining elevated for 25–35 days (mean concentration of progesterone ± SEM; 16.2 ± 2 ng/mg Cr). Initiation of aggression and sexual behavior by removed females increased significantly when they were isolated from the parent colony. The results demonstrated that odor cues from the complete colony in conjunction with behavioral / tactile / vocal cues from the nonbreeding colony members were not the major cues maintaining reproductive suppression in nonbreeding female naked mole-rats. Instead, our results suggest that female reproductive suppression in naked mole-rats is caused by a dominance-related behavioral mechanism requiring direct contact with the breeding female.     

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

The objective of this study was to determine if sexual swellings in mandrills (Mandrillus sphinx) are a reflection of reproductive endocrine state. Urine samples were assayed using an enzyme immunoassay measuring pregnanediol‐3‐glucuronide (PdG) and estrone conjugates (E₁C). Hormone patterns of ovarian cycles, pregnancy and lactation were characterized and compared with sexual swellings and copulations relative to menses and peak E₁C. Cycle lengths averaging 28.7 days and pregnancy length of 181 days determined by hormonal and sexual swelling measures were similar to those reported in other Old World primate species. First day of copulation was observed during rising E₁C concentrations and preceded observations of peak swelling by 1–2 days. Observations of peak sexual swellings occurred at or on the day after peak E₁C and decreased following the ovulatory increase in PdG. Observations of menses and sexual swellings are a useful method to track mandrill ovarian cycles and can assist zoos in determining the reproductive state of females in their collections. Zoo Biol 27:320–330, 2008. © 2008 Wiley‐Liss, Inc.     

The effect of hormonal stimulation on steroid levels in tissue incubates of the sterlet (<Emphasis Type="Italic">Acipenser ruthenus</Emphasis> L.)

Sex steroid and cortisol levels were compared in Leibovitz’s L-15 cultural medium after incubation of tissue samples from the intact female and male sterlet (Acipenser rhutenus L.) and from the fishes exposed to hormonal stimulation of sexual maturation by a superactive analog of mammalian gonadotropin-releasing hormone (LH-RH-A). A higher level of 17,20β,21-trihydroxy-4-pregnen-3-one (20βS) was revealed in the cultural medium following incubation of ovarian follicles isolated from females 5 h after LH-RH-A stimulation, as compared to the data obtained for intact females. The addition of 1 μM progesterone (P₄) to the cultural medium before incubation of ovarian follicles also led to an increase in the 20βS level in the incubates by the end of the experiment. Cortisol and testosterone levels in the incubates exhibited the same tendency. The blood cortisol level significantly increased in females 5 h after their hormonal stimulation. In males, showing high levels of androgens (testosterone and 11-ketotestosterone) in the blood serum before hormonal stimulation, high levels of these hormones were also detected in the cultural medium after incubation of testicular and liver tissue samples. The addition of P₄ to the medium before incubation stimulated the production of sex steroids and cortisol by the gonadal fragments. Upon addition of P₄ to the incubation mixture, containing liver tissue samples, the 20βS level increased.     

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.     

Age‐specific changes in sperm levels among female tsetse (Glossina spp.) with a model for the time course of insemination

The age, insemination and ovulation status of tsetse flies Glossina pallidipes Austen (n = 154369) and Glossina morsitans morsitans Westwood (n = 19659), captured over 11 years in Zimbabwe, are assessed by ovarian dissection. Instantaneous rates of insemination increase exponentially with age in both species; 90% insemination levels are reached after 5 days post‐emergence in G. m. morsitans and 7 days in G. pallidipes, varying little with season. More than 95% of both species have ovulated by the age of 8 days and 99% by 12 days. Older flies that have not ovulated are > 100‐fold more likely to be caught in October and November than in other months. A 500‐fold decrease in trap catches did not result in any detectible decrease in the probability of females being inseminated. The proportion of partially filled spermathecae rises for approximately 6 days then declines, consistent with some flies having mated more than once. For flies caught on electric nets, with wings undamaged during capture, wing‐fray data are used to extend ovarian age estimates up to 11 ovulations. Among these flies, the volume of sperm in the spermathecae declines little in flies that have ovulated up to seven times; thereafter, it declines by approximately 1% per ovulation. The time course of insemination and the mating frequency of females are important considerations in modelling tsetse fly populations, as well as for the dynamics of interventions involving the release of genetically‐modified insects, which should not be seriously compromised by the limited levels of polyandry currently observed.     

Serum 17β-estradiol and progesterone associated with mating behavior during early pregnancy in female rhesus monkeys

The phenomenon of postconception mating behavior was examined in a social group of rhesus monkeys living in an outdoor compound. Periodic blood samples and daily vaginal swabs were obtained from nine females beginning several weeks prior to conception and continuing through 6 weeks of pregnancy to permit an assessment of ovarian hormonal events associated with mating during early pregnancy. Each of the females showed a discrete period of copulatory activity during the periovulatory period which ceased within several days after the 17β-estradiol (E₂) ovulatory peak. In agreement with earlier reports, only a percentage of subjects (44%) exhibited a period of postconception mating, with copulatory activity beginning 19.8 (± 1.9) days following the E₂ peak and continuing for 9.5 (± 1.3 days). Implantation bleeding was detected in all of the subjects with the onset 19.5 (± 0.68) days after the E₂ peak. The interval between the E₂ peak and the onset of implantation bleeding was similar for all females. However, the duration of implantation bleeding was significantly shorter in females who exhibited postconception mating. The females who displayed postconception copulatory activity had significantly lower mean serum progesterone concentrations (2.33 ± 0.24 ng/ml vs. 3.64 ± 0.37 ng/ml) during the period associated with implantation bleeding and copulatory behavior. Although both groups had elevated concentrations of serum E₂ during this period, levels in the females who displayed postconception mating were significantly lower (173.8 ± 19.2 pg/ml vs 223.9 ± 28.8 pg/ml). These data demonstrate that the occurrence of postconception mating behavior in this environment is associated with a distinct pattern of ovarian hormonal events, and analysis suggests that differences in steroid concentrations probably account for the observed differences in implantation bleeding and copulatory behavior during pregnancy.     

Hormonal regulation of agonistic and affiliative behavior in female mongolian gerbils (Meriones unguiculatus)

Ovarian steroids and oxytocin (OT) have been implicated in the regulation of social behaviors. The purpose of the present study was to examine hormonal substrates of aggression and affiliation in the female Mongolian gerbil (Meriones unguiculatus), a highly social, monogamous rodent. Sexually naive adult females were paired with sexually experienced males for 48 h and their interactions videotaped. Females were gonadally intact and tested during vaginal estrus (INT) or ovariectomized and observed after the following treatments, administered by means of sc injections: EBEB (7 days of estradiol-benzoate); EBP (2 days of EB followed by progesterone), SALEB (saline, days 1-5 then 2 days of EB), OTEB (OT for days 1-5 then 2 days of EB); OTOIL (OT for days 1-5 then 2 days of OIL); or SALOIL (saline days 1-5 then 2 days of OIL). During the first hour of pairing INT females displayed higher levels of affiliation and lower levels of sniffing and agonistic behavior than SALOIL females. All hormonal treatments reduced agonistic behaviors when compared to SALOIL, although none of the hormonal treatments restored affiliation to INT levels. During the 48-h test overt aggression varied by treatment with INT, EBEB, EBP, and OTEB females displaying lower levels than SALOIL, while all groups displayed similar levels of affiliation. The results indicate that OT and E play a significant role in regulating male-directed aggressive behavior in females and that the presence of ovarian hormones as well as OT can increase affiliation during initial contact. Over a sustained period of cohabitation social cues appear to be more important in regulating affiliation than gonadal hormones.     

Zygoparity and sex steroid hormone profiles in bluemouth Helicolenus dactylopterus

Two hundred and seven individuals (103 females and 104 males) of bluemouth Helicolenus dactylopterus (Scorpaeniformes, Sebastidae), a commercially important deep‐water species with an unusual reproductive strategy, from the eastern Atlantic Ocean ranging from 13·9 to 37·5 cm total length (L_T) were analysed from September 2011 to October 2012. The analysis included gonad maturity phases and blood‐plasma levels of oestradiol‐17β (E₂), 11‐ketotestosterone (11‐KT) and 17,20β‐dihydroxypregn‐4‐en‐3‐one (17,20β‐P). Results confirmed the existence of an annual reproductive cycle with asynchrony between females and males and a spawning season from January to May. A pronounced peak in 17,20β‐P in October for both sexes was associated with possible mating behaviour and recent copula. Levels of E₂ increased preceding the elevation of the gonado‐somatic index during ovarian growth and were lower during regression and regeneration. The frequency distribution of oocyte–embryonic stages and variation of hormone levels suggest the existence of daily rhythms. Fertilization was detected between 2000–0000 and 0800–1200 h and spawning took place throughout the day peaking between 2000 and 0000 h. The cyclic pattern of sex steroids and ovarian recruitment provides a new insight into the reproductive strategy of this species.     

Progestin implants

Progestin implants for contraception are highly effective, safe, and the most convenient choice for many women. Progestin implants currently on the market, preparing for launch, or under investigation are reviewed here. Their basic galenic and pharmacokinetic features, as well as their contraceptive effectiveness, are described. The first progestin-only contraceptive implant placed on the market was Norplant, a multiunit system. Since then, several single- and double-rod implants have been developed, each using one of four different progestins: levonorgestrel, etonogestrel, Nestorone and nomegestrol acetate. Jadelle is similar to Norplant but consists of only two, rather than six, Silastic rods to simplify insertion and removal; nevertheless, levonorgesterel serum levels are identical, and performance is the same for both systems. The single implant systems reviewed here are: Implanon with a 3-year duration; Nestorone implants for breast feeding and non-breast feeding women lasting up to 2 years; and Uniplant, which is effective for 1 year. The advantages and disadvantages of progestin implants, the importance of counseling for increasing user satisfaction, and the future outlook for this contraceptive method 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.     

The study of renin–angiotensin–aldosterone in experimental diabetes mellitus

It is generally accepted that hypertension and other vascular pathologies increase in diabetes mellitus (DM) patients as a result of the renin–angiotensin–aldosterone (RAA) system. In this study, changes in the renin‐angiotensin‐aldosterone (RAA) system level was determined in Streptozotocin (STZ)‐injected rats. A total of 46 female Wistar albino rats (180–220 g body weight) was utilized in these experiments. STZ was given intraperitoneally to induce diabetes in rats. Streptozotocin (60 mg kg⁻¹ body weight) was dissolved in 0·1 m citrate–‐phosphate buffer (pH 4–5). The non‐diabetic rats were injected with sterilized buffer alone to act as a control group. Blood glucose levels were 398±8·2 mg dl⁻¹, 488±11·75 mg dl⁻¹ and 658±29·6 mg dl⁻¹ at days 3, 12 and 30 respectively. The level of plasma renin activity (PRA) was measured as 7·69±1·07 ng ml⁻¹ h⁻¹; 1·82±0·22 ng ml⁻¹ h⁻¹ and 0·67±0·12 ng ml⁻¹ h⁻¹ at days 3, 12 and 30, respectively. These values showed that the PRA levels are decreased with increased time period. Serum angiotensin converting enzyme (ACE, E.C. 3.4.15.1) levels were increased at days 12 and 30 (p<0·05 and p<0·005), whereas serum aldosterone levels were increased at days 3 and 12 (p<0·05). The level of urea and creatinine increased at days 12 and 30 (p<0·05 and p<0·005, respectively) when compared to the control group. The data from these experiments indicate that the PRA level decreased whereas ACE activity level increased in diabetic rats compared with the control. Aldosterone levels increased at the first stage of the experiment, but then decreased by the end of the experiment as a result of changes in renin and ACE levels. Copyright © 1999 John Wiley & Sons, Ltd.