Fecal Androgen Concentrations Research Articles

Deslorelin subcutaneous implants in Oryx dammah males for reproductive control

The aim of this study was to assess the effects of the deslorelin subcutaneous implant as a temporary contraceptive method in the Oryx dammah male. For this purpose, deslorelin at different doses, i.e. 14.1 mg and 9.4 mg, was subcutaneously implanted in three males (Phase 1) and one male (Phase 2) adult Oryx dammah, respectively. Quantitative behavior evaluation and androgen concentrations in feces and plasma were assessed before and after implant application. Fecal androgen concentrations observed in treated males were compared with those measured in one orchiectomized male and two females. Fecal androgen concentrations increased up to 15 days after the implant application, then progressively decreased, reaching the basal level at day150 in Phase 1. In Phase 2, levels remained high until day 60 and returned to basal level on day 120. Plasma testosterone concentration was higher on the day of implant application than three months later, but with variable ranges among males. A general increase of activity levels and hierarchical changes were observed after treatment, in accordance with hormonal variations. Despite males cohabiting with two fertile females during the observation period, no births were recorded. However, between the end of Phase 1 and the beginning of Phase 2, i.e. about 10–11 months after the first deslorelin implant, a fertile mating occurred leading to the birth of a calf. Therefore, we can hypothesize a contraceptive effect up to 10 months after the implant. Testicular histology performed on one male at the end of the Phase 2 showed no spermatogenetic activity.Our results suggest that deslorelin implant can be used to temporarily control reproduction in the Oryx dammah male. Behavior and fecal androgen measurements were useful and repeatable, non-invasive methods to monitor response.

Use of anti-gonadotropin-releasing hormone vaccines in African elephants (Loxodonta africana): A review

Background: Androgen-related aggressive behaviour and musth cause serious problems in captive African elephant bulls and often lead to human and animal injuries, and damage to property.Objectives: To review the work carried out with anti-gonadotropin-releasing hormone (GnRH) vaccines to control androgen-related behaviour and fertility in captive and free-ranging elephant bulls and the induction of anoestrus in elephant cows.Method: In the first study, an anti-GnRH vaccine from Pepscan was tested in six bulls (four captive and two free-ranging). Once the vaccine Improvac® became available, the effect on behaviour, the reproductive organs and semen quality was tested. Improvac® was also used to attempt induction of anoestrus in elephant cows.Results: The first study proved that aggressive behaviours are significantly associated with increased faecal androgen concentrations. Musth (n = 1) and aggressive behaviour (n = 2) were down regulated and correlated with a decline in faecal androgen concentrations. Aggression and musth could be controlled with Improvac® (600 µg), but were more consistent when the dose was increased to 1000 µg administered every five to six months. The same dose down regulated testicular function and bulls (n = 17) were rendered infertile within 12 months after commencement of treatment. Initial attempts to induce anoestrous with 600 µg in free-ranging elephant cows gave inconclusive results, but 1000 µg in captive cows delivered five-monthly was successful.Conclusion: The treatment of elephant bulls with Improvac® resulted in the successful down-regulation of androgen-related behaviour and sperm production in captive and wild elephant bulls of various ages (≤ 34 years). Preliminary studies to induce anoestrus in cows with Improvac® appear to be successful.

Urofecal steroid profiles of captive Blue-fronted parrots (Amazona aestiva) with different reproductive outcomes

Despite Psitaciformes (parrots) being the third largest nonpasserine order (398 species), it currently ranks second in number of threatened species (28%) according to the Internatinal Union for Conservation of Nature (IUCN) criteria. Since most of the literature concerning reproductive endocrinology in avian species derives from domestic and song birds, it is puzzling that advances in reproductive science for the Psitaciformes order lags far behind, in spite of the growing threats against them. In order to expand our knowledge of Neotropical parrots (Psittacidae), we examined annual changes in urofecal sex steroid metabolites of Blue-fronted amazon pairs (Amazona aestiva) exhibiting successful (nestlings) and unsuccessful breeding (infertile or no eggs). Urofecal samples were collected over a year from eight breeding pairs housed under the same environmental and management conditions. Fecal androgen and progestagen concentrations were determined in males and females, respectively, by enzyme immunoassays previously validated for this species. All eggs were registered between late winter and mid-spring, and egg-laying intervals varied between females (range: 1–8 days; average 3.60 ± 0.51 days). Similar profiles of urofecal progestagens were observed in reproductively successful females and females producing infertile eggs, with progestagen peaks preceding egg laying events (1.77 ± 0.50 days). In contrast, non-laying females had no rises in progestagens during the year. Successful and unsuccessful males did not displayed distinct annual patterns of androgen production, and apart from the peaks during the breeding season, more than half of the individuals intriguingly presented significant increases from late summer to early autumn, a period without reproductive activity. Finally, we noticed that samples with progestagen levels exceeding 40 ng/g had very high probability (>97.5%) to be from females in pre-laying or laying phases, suggesting a feasible application of this characteristic to noninvasively discriminate the reproductive status in amazon females with an accuracy and sensitivity of 94.55% and 58.13%, respectively. Our findings confirmed that urofecal progestagens and androgens are good indicators of the gonadal condition in Blue-fronted amazons, but there is still much to be done for their extensive use in artificial insemination or selection of the most suitable breeding birds for the season.

Non‐invasive physiological markers demonstrate link between habitat quality, adult sex ratio and poor population growth rate in a vulnerable species, the Cape mountain zebra

Abstract Effective conservation and species management require an understanding of the causes of poor population growth. Conservation physiology uses biomarkers to identify factors that contribute to low individual fitness and population declines. Building on this, macrophysiology can use the same markers to assess how individual physiology varies with different ecological or demographic factors over large temporal and spatial scales. Here, we use a macrophysiological approach to identify the ecological and demographic correlates of poor population growth rates in the Cape mountain zebra metapopulation. We use two non‐invasive biomarkers: faecal glucocorticoids as a measure of chronic stress, and faecal androgens as an indicator of male physiological status. We found that faecal glucocorticoid concentrations were highest in the spring prior to summer rainfall, and were elevated in individuals from populations associated with low‐quality habitat (lower grass abundance). In addition, faecal androgen concentrations were higher in populations with a high proportion of non‐breeding stallions (where male:female adult sex ratios exceed 2:1) suggesting sex ratio imbalances may intensify male competition. Finally, population growth rate was negatively associated with faecal glucocorticoid concentrations and female fecundity was negatively associated with faecal androgens, indicating a relationship between hormone profiles and fitness. Together, our results provide cross‐population evidence for how poor population growth rates in Cape mountain zebra can be linked to individual physiological biomarkers. More broadly, we advocate physiological biomarkers as indicators of population viability, and as a way to evaluate the impact of variable ecological and demographic factors. In addition, conservation physiology can be used to assess the efficacy of management interventions for this subspecies, and this approach could inform models of species’ responses to future environmental change. A plain language summary is available for this article.

Androgens predict parasitism in female meerkats: a new perspective on a classic trade-off

The immunocompetence handicap hypothesis posits that androgens in males can be a 'double-edged sword', actively promoting reproductive success, while also negatively impacting health. Because there can be both substantial androgen concentrations in females and significant androgenic variation among them, particularly in species portraying female social dominance over males or intense female-female competition, androgens might also play a role in mediating female health and fitness. We examined this hypothesis in the meerkat (Suricata suricatta), a cooperatively breeding, social carnivoran characterized by aggressively mediated female social dominance and extreme rank-related reproductive skew. Dominant females also have greater androgen concentrations and harbour greater parasite loads than their subordinate counterparts, but the relationship between concurrent androgen concentrations and parasite burdens is unknown. We found that a female's faecal androgen concentrations reliably predicted her concurrent state of endoparasitism irrespective of her social status: parasite species richness and infection by Spirurida nematodes, Oxynema suricattae, Pseudandrya suricattae and coccidia were greater with greater androgen concentrations. Based on gastrointestinal parasite burdens, females appear to experience the same trade-off in the costs and benefits of raised androgens as do the males of many species. This trade-off presumably represents a health cost of sexual selection operating in females.

Reproductive seasonality and sperm cryopreservation in the male tufted deer (Elaphodus cephalophus)

The tufted deer is a small deer, listed as near threatened on the International Union for Conservation of Nature Red List, and there is no information on the fundamental reproductive biology of this species. In this study, we report for the first time, characterization of male reproductive traits and cryopreservation of semen in this species. Males were subjected to electroejaculation during each season (autumn, winter, spring, and summer), and ejaculates were assessed for motility and quality traits. Fecal samples were collected 3 to 5 times weekly for 2 years and analyzed for androgen metabolites using enzyme immunoassay. Ejaculates with greater than 70% motility were cryopreserved using Beltsville extender (BF5F) or Triladyl. Straws were thawed and assessed subjectively as well as swim-up processed to isolate motile spermatozoa for computer-assisted sperm analysis and acrosome integrity at hourly interval. Tufted deer male reproductive and semen traits peaked in autumn. Mean fecal androgen concentrations were highest in the summer compared with baseline values during rest of the year. Sperm motility and acrosome integrity were lower immediately after thawing in both cryodiluents compared with raw ejaculates. Motility characteristics after swim-up were higher in BF5F compared with Triladyl. Four hours after thawing, both percent sperm motility and progression decreased further and were similar between BF5F and Triladyl. However, the proportion of spermatozoa with intact acrosomal membranes was higher in BF5F than Triladyl. Results indicate that tufted deer exhibit seasonal variations in reproductive traits and that BF5F better preserves sperm motility and acrosomal integrity after cryopreservation compared with Triladyl.

Season- and Age-related Reproductive Changes Based on Fecal Androgen Concentrations in Male Koalas, <i>Phascolarctos cinereus</i>

The purposes of the present study were to clarify age- and season- related androgen patterns, and to compare the reproductive physiology between Japanese captive koala populations and Australian populations. To measure fecal androgens, feces were collected from male koalas (4.2 to 13.8 years of age) kept in Japanese zoos. Fecal androgens were extracted with methanol from the lyophilized samples and determined by enzyme immunoassay using 4-androstene-3,17-dione antibody. Fecal androgen concentration in male koalas increased after sexual maturation and remained relatively high until old age. In the survey with the Japanese zoo studbook of koalas, copulation (conception) month showed a pyramid shape with a peak in March to June (60.7%) in koalas born and reared in Japanese zoos and from July to April with the highest concentration in September to January (69.7%) in Australian institutes. Japanese zoo koala populations have a characteristic physiological cycle adapted to Japan's seasonal changes. The suitable month of year for copulation or conception in Japan is diametrically opposed to that in Australia. Mean fecal androgen concentrations by month in the males born and reared in Japan indicated annual changes with the highest concentration in May and the lowest value in November. Fecal androgen analysis may be a noninvasive alternative tool to monitor circulating testosterone and may be helpful in understanding reproductive activity and physiology in male koalas.

Protracted Reproductive Seasonality in the Male Giant Panda (Ailuropoda melanoleuca) Reflected by Patterns in Androgen Profiles, Ejaculate Characteristics, and Selected Behaviors1

The female giant panda (Ailuropoda melanoleuca) experiences a brief (24-72 h) seasonal estrus, occurring once annually in spring (February-May). Our aim was to determine the existence and temporal profile of reproductive seasonality in the male of this species. The study was facilitated by 3 yr of access to eight giant panda males living in a large breeding center in China. Seasonal periods for the male were defined on the basis of female reproductive activity as prebreeding, breeding (early, peak, late), and nonbreeding seasons. Testes size, fecal androgen excretion, ejaculated sperm density, and frequency of reproductive behaviors (i.e., locomotion, scent marking, vocalizations) increased (P < 0.05) from the prebreeding period (October 1-January 31) to the early breeding season (February 1-March 21). Testes volume and sperm concentration were maximal from March 22 through April 15, a period coinciding with maximal female breeding activity. The occurrence of male reproductive behaviors and fecal androgen concentrations began declining during peak breeding and continued from April 16 through May 31 (late breeding period), returning to nadir throughout the nonbreeding interval (June 1-September 30). Reproductive quiescence throughout the latter period was associated with basal testes size/volume and aspermic ejaculates. Our results reveal that testes morphometry, fecal androgen excretion, seminal quality, and certain behaviors integrated together clearly demonstrate reproductive seasonality in the male giant panda. The coordinated increases in testes size, androgen production, sperm density, and sexual behaviors occur over a protracted interval, likely to prepare for and then accommodate a brief, unpredictable female estrus.

Annual profile of fecal androgen and glucocorticoid levels in free-living male American kestrels from southern mid-latitude areas

Fecal samples and behavioral data were collected at a fortnightly basis during 11 months period from free-living male American kestrels living in southeast Brazil (22°S latitude). The aim was to investigate the seasonal changes in testicular and adrenal steroidogenic activity and their correlation to reproductive behaviors and environmental factors. The results revealed that monthly mean of fecal glucocorticoid metabolites in May and June were higher than those estimated in November. In parallel, monthly mean of androgen metabolites in September was higher than those from January to April and from October to November. Molt took place from January to March, whereas copulation was observed from June to October but peaked in September. Nest activity and food transfer to females occurred predominantly in October, and parental behavior was noticed only in November. Territorial aggressions were rare and scattered throughout the year. Multiple regression analysis revealed that fecal androgen levels are predicted by photoperiod and copulation, while fecal glucocorticoid levels are only predicted by photoperiod. Bivariate correlations showed that fecal androgen metabolites were positively correlated with fecal glucocorticoid metabolites and copulation, but negatively correlated with molt. Additionally, copulation was positively correlated with food transfer to females and nest activity, but negatively correlated with molt. These findings suggest that male American kestrels living in southeast Brazil exhibit significant seasonal changes in fecal androgen and glucocorticoid concentrations, which seem to be stimulated by decreasing daylength but not by rainfall or temperature.

Longitudinal monitoring of plasma and fecal androgens in the Tasmanian devil ( Sarcophilus harrisii) and the spotted-tailed quoll ( Dasyurus maculatus)

Improved knowledge of the breeding biology of carnivorous marsupials is warranted given their heightened conservation status. Past studies have focused on smaller dasyurids and little is known of male reproductive physiology in the larger species. This study aimed to characterize the pattern of androgen concentrations in male devils and spotted-tailed quolls and to evaluate fecal steroid measurement as a practical, alternative technique for monitoring reproductive activity. Blood and fecal samples were collected from captive adult devils ( n = 6) and adult quolls ( n = 8). Plasma and fecal androgen concentrations were significantly positively correlated. In both species there was a significant effect of season on androgen concentrations; and the annual increase preceded female estrus activity. For devils, fecal androgens were elevated during the austral summer: peak concentrations were observed in January–February, and copulation occurred from late February–late May. In quolls, fecal androgen concentrations were highest during austral autumn/winter: the annual increase began in April and copulation occurred from mid-May to early October. The lengthy period of elevated plasma and fecal androgens and protracted annual period of mating activity implies a period of extended spermatogenesis in both species.

Social correlates of testosterone and ornamentation in male mandrills

We investigated relationships between fecal androgen concentrations, facial coloration and behaviour in semi-free-ranging male mandrills. We found that fecal androgen levels were significantly positively related to dominance rank, independent of rank stability and the mating period, suggesting that male mandrills live in a permanently aggressive context in which they must actively maintain their dominance status. Facial red coloration was also significantly related to both fecal androgen levels and rank, with high ranking males having both higher androgen levels and redder faces, although dominant males did not always have the highest androgen levels or the reddest faces. Predictive relationships between androgen levels, coloration and rank were short-term. Androgen concentrations and facial redness both increased in the presence of receptive females, as did the former during periods of rank instability. We conclude that male facial redness is likely to represent an honest signal (to other males) of current androgen status, competitive ability and willingness to engage in fights and that females may also use this to assess male condition. Further, our findings provide support for the “challenge hypothesis” as originally proposed for birds by Wingfield.

Faecal androgen concentrations in adult male spotted hyaenas, Crocuta crocuta, reflect interactions with socially dominant females

Androgens influence many physiological processes in male vertebrates and have been linked to variation in aggression and reproductive behaviour in many species. This study examined relationships between social behaviour and androgens excreted by adult male spotted hyaenas. In this species, males rarely direct reproductive aggression towards other males, but they actively court females that are socially dominant to them. We tested whether intersexual interactions were associated with elevated androgens in adult males. We combined behavioural data collected over 7 years from one large clan in Kenya with analysis of faecal androgens (fA) to determine which of the following were the best predictors of fA concentrations in males: social rank, tenure in the clan, courtship, association with females in particular reproductive states, reproductive aggression among males or nonreproductive aggression among males. We found no systematic influence of age or social status on fA in either adult natal or immigrant males. Concentrations of fA were greater in immigrants when they courted females than at other times, and greater fA concentrations were also associated with male–male aggression involving defence of a female but not with male–male aggression occurring in other contexts. fA concentrations in immigrants were also positively correlated with degree of association with females near the time of conception, but not with lactating females. These findings suggest that androgen concentrations in immigrant male hyaenas are more related to interactions with attractive females than to competitive male interactions. The most important challenge to adult males may be interacting with attractive females to which males are socially subordinate.

Seasonal variation in the endocrine–testicular function of captive jaguars ( Panthera onca)

Captive adult male jaguars ( Panthera onca) from two locations in southeast Brazil were studied to evaluate the effects of season on endocrine and testicular function. For assessment of testicular steroidogenic activity, androgen metabolite concentrations were measured in fecal samples collected one to three times per week over 14 ( n=4), 9 ( n=1) or 7 months ( n=1). To assess seasonality, data were grouped by season (summer: December–February; autumn: March–May; winter: June–August; spring: September–November). Additionally, samples collected in the dry season (March–August) were compared with those collected in the wet season (September–February). There were no differences ( P>0.05) in fecal androgen concentrations in samples collected in spring, summer, autumn, and winter (480.8±50.4 ng/g, 486.4±42.0 ng/g, 335.4±37.7 ng/g, and 418.6±40.4 ng/g dry feces). However, there were differences ( P<0.05) in fecal androgen concentrations between the dry and wet seasons (380.5±28.0 ng/g versus 483.9±32.3 ng/g dry feces). Sperm samples, collected from all males twice (approximately 6 months apart) were similar; mean (±S.E.M.) motility, concentration and morphology were 57.0±4.5%, 6.3±2.4×10 6 ml −1, and 60.8±3.1%, respectively. In conclusion, androgen metabolite concentrations in the captive male jaguar were not affected by season, but there was a difference between the wet and dry periods. Further research is needed to verify these results.

Non-invasive monitoring of fecal androgens in spotted hyenas ( Crocuta crocuta)

Spotted hyenas ( Crocuta crocuta) exhibit an array of behavioral and morphological characteristics that set them apart from other mammals: females are heavier and more aggressive than males, and females have external genitalia that closely resemble those of the male. Because androgenic hormones might mediate the expression of these traits, androgens are of great interest in this species. Past work on circulating androgens in wild hyenas has been limited, in part because of small sample sizes. In this study we validated a non-invasive method of monitoring variation in androgens by measuring total androgen metabolites in the feces of wild and captive spotted hyenas with an enzyme immunoassay. HPLC analysis revealed multiple immunoreactive androgen metabolites in fecal extracts from both males and females. LHRH challenge in three male and two female hyenas in captivity caused an increase in fecal androgens one to three days after LHRH injection. Furthermore, presence of bone in the diet did not affect fecal androgen concentrations in captive female hyenas. In wild spotted hyenas, time of day of fecal deposition, time elapsed between deposition and freezing of the sample, and time elapsed between freezing and extraction did not systematically affect fecal androgen concentrations. Finally, in wild hyenas, fecal androgen patterns mirrored plasma testosterone patterns in that adult immigrant males had higher concentrations than adult natal males, and pregnant females had higher concentrations than lactating females. These methods can therefore be used in future studies addressing relationships among fecal androgens, social status, reproductive state, and behavior in spotted hyenas.

Seasonal analysis of semen characteristics, serum testosterone and fecal androgens in the ocelot ( Leopardus pardalis), margay ( L. wiedii) and tigrina ( L. tigrinus)

Captive adult male ocelots ( Leopardus pardalis, n=3), margays ( L. wiedii, n=3) and tigrinas ( L. tigrinus, n=4) in two locations in southern Brazil were studied for 14 consecutive months to evaluate the effect of season on testicular function. Reproductive evaluations, including testicular measurements, electroejaculation and blood collection were conducted monthly. Fecal samples were collected weekly for androgen metabolite analysis to assess testicular steroidogenic activity. Ocelots had the highest number of motile spermatozoa in the ejaculate (114.7±15.8×10 6; P<0.05), the highest percentage of morphologically normal spermatozoa (82.4±1.2%; P<0.05) and the highest concentration of fecal androgens (1.71 vs. 0.14 μg/g; P<0.05). Margays and tigrinas had lower numbers of motile spermatozoa (23.4±2.8×10 6, 74.2±8.9×10 6, respectively), lower percentages of morphologically normal spermatozoa (57.4±2.8, 59.2±3.5%, respectively), and lower fecal androgen concentrations (0.15±0.01, 0.23±0.01 μg/g, respectively). Serum testosterone concentrations were similar among the three species. Fecal androgen concentrations were not affected by season, with the exception of the ocelot where concentrations were higher ( P<0.05) in the summer. Ejaculates were collected throughout the year; however, peaks in average sperm production were observed during the summer for all species. In summary, this study has identified several species differences in male testicular traits among ocelots, margays and tigrinas. Results of longitudinal reproductive assessments suggest males of each species are capable of breeding throughout the year.

Fecal androgen metabolite analysis for noninvasive monitoring of testicular steroidogenic activity in felids

Limited data are available on long-term, seasonal changes in testicular steroidogenic activity in nondomestic felids, primarily because of the difficulties associated with longitudinal blood sampling (e.g., handling, restraint, anesthesia). Therefore, a noninvasive approach for assessing testicular androgen production was developed using the domestic cat (Felis catus) as a model. Two adult males were injected i.m. with 4 μCi14-testosterone to determine the time course and relative proportions of androgen metabolites excreted in urine and feces. Peak urinary radioactivity was detected 13 and 19 hr postinjection and accounted for ∼8% of the total radioactivity recovered. High performance liquid chromatography (HPLC) analysis detected multiple polar urinary metabolites, none of which eluted with the 3H-testosterone reference tracer. The majority of urinary testosterone metabolites consisted of nonenzyme-hydrolyzable, water-soluble (presumably conjugated) forms. In feces, radioactivity was detected in the first sample collected at 22 hr postinjection for both males, although peak metabolite excretion in one male was not observed until 61 hr postinjection. HPLC analysis detected several fecal metabolites consisting primarily of nonhydrolyzable, water-soluble forms (84.4 ± 0.9%) with some ether-soluble forms (15.6 ± 0.9%). None of the fecal androgen metabolites were associated with free testosterone. However, one or more of the water-soluble fecal metabolites was quantifiable using a commercially available testosterone radioimmunoassay. The biological relevance of this immunoactivity was confirmed in the domestic cat; concentrations were high in adult, intact males and nondetectable in intact females and castrated males and females. In addition, fecal androgen concentrations in a male Pallas' cat (Felis manul) exhibited seasonal fluctuations that corresponded with parallel changes in serum testosterone and ejaculate quality. These data indicate that testicular steroidogenic activity can be monitored non invasively in felids, providing a potentially valuable tool for endangered felid management to: (1) assess pubertal status, (2) determine the influence of season on reproduction, and (3) diagnose possible causes of sub- or infertility. © 1996 Wiley-Liss, Inc.