Reciprocal interaction between seasonal testis and thyroid activity in Zembra Island wild rabbits (Oryctolagus cuniculus): effects of castration, thyroidectomy, temperature, and photoperiod

The reproductive cycle of wild rabbits (Oryctolagus cuniculus) living in Zembra Island (North Tunisia) is dependent on an external factor, the photoperiod: the gonads are inhibited by long days and stimulated by short days or melatonin implants. Here we studied the role of an internal factor, thyroid hormones and the possible thyroid-gonadal interrelationships, in animals captured on Zembra Island and maintained in natural conditions of photoperiod and temperature. We determined the seasonal profile of the thyroid and testis cycles and investigated the effects of castration and thyroidectomy on the seasonal testosterone and thyroxine cycles. Plasma thyroxine and testosterone levels followed similar, parallel seasonal patterns, with a peak in autumn (October) and low values from January to August. In thyroidectomized animals, plasma testosterone levels, although significantly higher than those in controls (P < 0.001), remained low throughout the 13 mo of the experiment, and no testicular reactivation was observed in the fall. In castrated animals, despite the increase in thyroxine concentration in the 3 mo following castration (P < 0.01), plasma thyroxine levels remained low during the 2 yr of the study. We then investigated the combined effects of long days (16L:8D) and moderately high temperature (25 degrees C) on these two endocrine axes. In constant gonado-inhibiting conditions (16L:8D), whether the temperature was kept constantly high or allowed to fluctuate naturally, no reactivation of the thyroid and testicular axes was observed in the fall. In control animals, the peaks of testosterone and thyroxine concentrations observed in September were larger (P < 0.001) than those in animals subjected to the same natural photoperiod conditions but with constantly high temperature. The lower level of autumnal testis stimulation (P < 0.001) in animals maintained in conditions of constant high temperature (25 degrees C) may be attributed to the low thyroxine levels induced by high temperature. These results clearly confirm that the thyroid and testicular cycles display similar seasonal variations and show that the thyroid and gonadal axes are strictly interdependent. This study provides the first demonstration, for a given species, that the seasonal reactivation of gonad activity is controlled by the thyroid, and thyroid activity is controlled by the gonads.     

Social cues and hormone levels in male Octodon degus (Rodentia): a field test of the Challenge Hypothesis

Social interactions are important factors determining and regulating individual behaviors. Testosterone has been related to agonistic interactions, while glucocorticoids have been related to social stress, especially during interactions of dominance. We compared testosterone and cortisol concentrations in male degus (Octodon degus, Rodentia) under laboratory conditions without male social interactions, with data from wild males in nature. Under natural conditions, males should present higher levels of testosterone during the breeding season due to social interactions (Challenge Hypothesis). Alternatively, intense social instability could act as a stressing environment, raising glucocorticoids, which inhibit testosterone concentrations. Our results show a significant increase in agonistic interactions between males during the breeding season, and disappearance of non-agonistic male interactions during this period. Hormone levels in breeding season show nonsignificant differences between laboratory groups, but testosterone concentrations in field males were significantly higher than in laboratory males. Testosterone levels were similar among pre-breeding and breeding periods, but in field animals the concentration was approximately 30% higher than in laboratory degus. In field animals, we found two different mating strategies: resident males, with territorial behavior, and transient males, displayed an opportunistic approach to females. Finally, cortisol presents a similar pattern in both laboratory and field animals; pre-breeding values of cortisol are higher than during the breeding season. This suggests that social interactions in O. degus activate a rise in testosterone, supporting the Challenge Hypothesis, and could be considered as partial support of the Social Stress Hypothesis.     

The effects of cortisol and testosterone on renal function in male Antechinus stuartii (Marsupialia)

Seasonal changes in the physiology of Antechinus stuartii result in complete male mortality after mating. The most important endocrine changes in males are large rises in plasma testosterone and cortisol concentrations. Glomerular filtration rate (GFR) in males declines coincident with high plasma testosterone and cortisol. In the present study GFRs were measured in males captured in May (when endogenous plasma testosterone and cortisol levels are low) and given depot injections of either saline, testosterone-only, cortisol-only or testosterone plus cortisol at doses designed to mimic plasma levels during the mating period. GFR decreased significantly with testosterone injection, independent of cortisol treatment. Urinary concentrations of sodium and chloride, and osmolality decreased significantly with cortisol treatment, although the addition of testosterone reversed the effect. Total urinary excretion of electrolytes was similar between groups. Plasma potassium levels significantly increased in testosterone plus cortisol treated males. Plasma sodium levels significantly increased and plasma chloride significantly decreased in all groups treated with cortisol. Water consumption significantly increased in all cortisol-treated males and food consumption significantly increased in all testosterone-treated males. The seasonal renal functional changes observed in A. stuartii were mimicked by testosterone administration. Accepted: 23 January 1998     

Primary and secondary stress responses to line capture in the blue mao mao

Primary and secondary stress responses were measured in wild Scorpis violaceus subjected to burst swimming from angling. Fish were blood sampled from 20 s to 30 min after hooking. Consequent rises in plasma adrenaline (14–316 nmol l⁻¹), noradrenaline (25–345 nmol l⁻¹), and cortisol (0.4–197 ng ml⁻¹) correlated with time since capture, and plasma lactate (0.1–12.2 mmol l⁻¹) reflected work done during intense exercise. Haemoglobin concentration and haematocrit also increased with exercise, and erythrocyte swelling occurred. Wild S. violaceus demonstrated a spontaneity and intensity of exercise not seen in fish acclimatized to aquarium conditions. By contrast, the stress responses of fish in captivity, despite careful husbandry, differed qualitatively and quantitatively from those in the wild. Cannulated fish had higher resting plasma cortisol concentrations (61.9±9.5 ng ml⁻¹) than did rapidly caught wild fish (<5 ng ml⁻¹) and these values were not significantly changed with burst swimming. Catecholamine secretion, possibly suppressed by cortisol, was insufficient to cause erythrocyte swelling. Erythrocyte nucleotides do not play a role in exercise, but are elevated in captive fish. It is hypothesized that primary endocrine responses are triggered by higher cortical processing of sensory information which is fundamentally different in the natural environment.     

Plasma leptin levels of elite endurance runners after heavy endurance training

A decrease in testosterone levels and an increase in cortisol levels are observed in male athletes with the overtraining syndrome (OTS). Cortisol causes blood leptin levels to rise and testosterone has an inverse relationship with blood leptin levels. Therefore, we hypothesized that the hormonal changes as a result of OTS induce an increase in leptin. To test this hypothesis, we examined the relationship among changes in leptin, testosterone and cortisol in thirteen male collegiate distance runners (aged 20.3+/-1.1 years) before and after an 8-day strenuous training camp. Runners ran 284.1+/-48.2 km during the training camp. Body fat percentages and plasma glucose concentrations decreased significantly after the training. Non-ester fatty acids and total cholesterol concentrations in blood were unchanged. Serum cortisol concentrations showed a significant increase after the training camp (from 11.82+/-2.00 microg/dl to 16.78+/-3.99 microg/dl), and serum testosterone decreased significantly (from 408.0+/-127.6 ng/dl to 265.2+/-97.6 ng/dl). The ratio of testosterone to cortisol (TCR) dropped by 50% after training (from 35.62+/-13.69 to 16.94+/-8.47). These results suggest that the subjects reached a state of the OTS. Contrary to our hypothesis, plasma leptin was not significantly changed (from 1.34+/-0.29 ng/ml to 1.49+/-0.18 ng/ml). Delta Plasma leptin was not significantly correlated with delta serum cortisol, delta TCR or delta fat percentage. However, delta serum testosterone was positively correlated with delta plasma leptin (r=596, p<0.05). Plasma leptin concentrations might modulate the secretion of testosterone in overtraining conditions. In conclusion, the change in blood leptin level is independent of the changes in cortisol, TCR and fat percentage in highly trained male athletes in the state of the OTS.     

Seasonal and social correlates of fecal testosterone and cortisol levels in wild male muriquis (Brachyteles arachnoides)

Fecal testosterone and cortisol levels were analyzed from six wild male muriquis (Brachyteles arachnoides) over a 19-month period at the Esta??o Biológica de Caratinga in Minas Gerais, Brazil, to investigate the hormonal correlates of seasonal sexual behavior and environmental conditions. Group mean testosterone levels based on weekly samples from the six males did not differ between copulatory and noncopulatory periods or between rainy and dry seasons. Cortisol levels did change with copulatory periods, and were significantly higher during the second dry season, when mating continued following an exceptionally heavy rainy season, than during the first dry season, when mating ceased. Males exhibited individual variation in the timing of their hormone shifts relative to their sexual activity, but neither hormone levels nor sexual activity were related to male age. Despite individual differences in the timing of testosterone fluctuations around the onset and offset of the copulatory season, all males exhibited elevated cortisol concentrations following a slight increase in testosterone at the beginning of the copulatory season. Both the lack of significant changes in testosterone levels with the onset of the rainy and copulatory season and the lack of prebreeding increases in cortisol may be related to the low levels of overt aggression displayed by male muriquis over access to mates.     

Circulating androgens are influenced by parental nest defense in a wild teleost fish

While social interactions influence vertebrate endocrine regulation, the dynamics of regulation in relation to specific behaviors have not been clearly elucidated. In the current study, we investigated whether androgens (testosterone) or glucocorticoids (cortisol) play a functional role in aggressive offspring defense behavior in wild smallmouth bass (Micropterus dolomieu), a teleost fish with sole paternal care. We measured circulating testosterone and cortisol concentrations in plasma samples taken from parental males following a simulated nest intrusion by a common nest predator, the bluegill sunfish (Lepomis macrochirus). To understand whether endocrine regulation changes across the parental care period, we looked both at males guarding fresh eggs and at males guarding hatched embryos. Plasma testosterone levels increased in males subjected to a simulated nest intrusion when compared to sham controls. Testosterone concentrations in males guarding embryos were lower than in males guarding fresh eggs, but circulating testosterone was positively correlated with the level of aggression towards the nest predator at both offspring development stages. However, there was no increase in cortisol levels following a simulated nest intrusion, and no relationship between cortisol and any measured parameter. These results suggest that androgens play an important role in promoting aggressive nest defense behavior in teleost fish.     

Origin of European rabbit (Oryctolagus cuniculus) in a Mediterranean island: Zooarchaeology and ancient DNA examination

Mammalian species presently living on Mediterranean islands have been brought in by man. The question of their geographical origin and of the time of their introduction is often a matter of debate. We studied this problem using a population of rabbits (European rabbit: Oryctolagus cuniculus) living in Zembra, an island off Tunisia. Archaeological surveys show that rabbit has been introduced to the island by Bronze Age or Roman people, between the IIIrd Millenium B.C. and the IIIrd century A.D. Part of the 16S-rRNA gene of mitochondrial DNAs from fossil bones of different ages (dated back to 130–390 A.D.) was characterized and compared to that of present day rabbits of differing geographical origin. The data suggest that animals present on Zembra in late Roman times belonged to the same maternal lineage as present populations from Northern Spain and Southern France.     

Effects of capture on adrenal steroid and vasopressin concentrations in free-ranging bottlenose dolphins (Tursiops truncatus)

Marine mammals are routinely caught in the wild in an effort to monitor their health. However, capture-associated stress could potentially bias various biochemical parameters used to monitor the health of these wild caught animals. Therefore, the effects of capture were quantified by measuring plasma adrenal steroids and arginine vasopressin (AVP) in free-ranging bottlenose dolphins (Tursiops truncatus) (n=31). Total capture and restraint times were also correlated to hormone concentrations to quantify the effects of capture. Significant, positive correlations between corticosterone and cortisol (R=0.752; P<0.0001), and between corticosterone and aldosterone (R=0.441; P=0.045) were demonstrated. Significant correlations between capture and restraint time and hormone levels were not observed. Animals restrained for less than 20 min exhibited hormone levels similar to those for animals restrained for more than 20 min. The positive correlations among the adrenal steroids suggest that release of these steroids was stimulated by adrenocorticotropin (ACTH). The lack of a correlation between cortisol and AVP indicates that AVP did not influence ACTH-induced cortisol release in this situation. The study suggests that (1) a typical hypothalamic-pituitary-adrenal axis is present in these animals, and (2) the relatively short capture and restraint times did not induce a significant neuroendocrine stress response.     

Responses of naïve and experienced European rabbits to predator odour

The response of prey species to predator scent has been investigated in many mammalian species; however, there is little information about the responses of European wild rabbits at the population level. Therefore, we conducted a simple experiment to investigate the behavioural response of a rabbit population to native predator cues in the wild. We compared the response to the scent of a predator (red fox) in a wild rabbit population bred in semi-natural conditions and naïve to terrestrial predators with the response of a population in a similar environment where terrestrial predators were present. The response to predators was based on rabbit abundance, inferred from pellet counts and measured by the defecation rate per day (DRD). Our results indicate that rabbits responded to the odour of fox faeces in the treatment warrens, resulting in a lower DRD. The main anti-predator behaviour observed was spatial avoidance (warren abandonment), which seemed to be more accentuated for rabbits who had not previously had contact with foxes in the plot where terrestrial predators were excluded. In both the fenced and the unfenced plot, the differences in the effect of the predator odour between the control and treatment warrens disappeared after cessation of treatment, suggesting a flexible and adaptive behaviour of rabbits to predator cues.

     

Long-day inhibition of reproduction and circadian photogonadosensitivity in Zembra Island wild rabbits (Oryctolagus cuniculus).

We investigated the effects of photoperiod on testicular activity in wild rabbits (Oryctolagus cuniculus) captured on Zembra Island (North Tunisia) and maintained in experimental photoperiodic conditions. Sexually inactive animals were subjected to alternate 3-mo periods of short days (8L:16D) and long days (16L:8D) for 1 yr. Testicular activity increased significantly and then decreased to levels equivalent to or lower than those measured during sexual quiescence after 1 mo of 8L:16D or 16L:8D, respectively. Eight groups of sexually active animals were also exposed to 8L:16D for 60 days. The light phase was divided into two photofractions (7.5 and 0.5 h). The short photofraction interrupted the dark phase 9.5-18.5 h after the beginning of the main photofraction. Testicular activity was inhibited if the short photofraction interrupted the dark phase 12.5 h or more after the beginning of the main photofraction. These results clearly confirm that photoperiod affects reproduction in this species: Short days stimulate reproduction, whereas long days inhibit it. The asymmetric pattern of skeleton photoperiods used demonstrated the existence of a circadian rhythm for photogonadosensitivity, with the photosensitive phase beginning 12.5 h after dawn. In this species, photoperiod length controls both the beginning and the end of the reproductive period. These results differ from those obtained with continental populations of wild rabbits, in which reproduction is inhibited by short day length. This difference may reflect genetic drift linked to the geographic isolation of this population, which is known to have been present on this small island for more than 2000 yr.     

The effect of unilateral castration on plasma and testicular testosterone in rabbits from birth to 60 days.

Male rabbits were hemigonadectomized every 10 days from 1 to 50 days of age and were sacrificed 10 days after the operation. Non-operated controls were sacrificed at the same stages. Plasma and testicular testosterone were quantified by radioimmunoassay. Compensatory testicular hypertrophy was not observed in hemicastrated rabbits. Hemicastration has variable effects on testicular and plasma testosterone levels as a function of age. When it is performed at birth, there is a significant increase in testicular and plasma concentrations of testosterone. Similar results are obtained if the operation is performed at 30 days. Unilateral castration performed at 10, 20 and 50 days reduces testicular and plasma testosterone levels. When hemicastration is performed at 40 days or at the adult age, testicular and plasma testosterone levels are not modified.     

Provision of artificial warrens as a means to enhance native wild rabbit populations: what type of warren and where should they be sited?

In Spain, wild rabbits are essential for some highly endangered species, and, therefore, many actions have been undertaken to increase their populations. In the present study, artificial warrens are provided as a means to increase shelter for native wild rabbit populations in a given area. We evaluate the use of three types of warrens by rabbits and the effect on that use of five habitat characteristics at two spatial scales (500 × 500-m grids and 25-m plots). To evaluate that use, we identified pre-established signs at the entrances to each warren, and based on this, we calculated occupancy rate and activity. Our results indicate that rabbit abundance within a grid is the only variable which simultaneously explains both the greater occupancy and the higher activity in the artificial warrens located in that grid. Some 73.2% of the grids showed signs of rabbit use at the time of the evaluation. However, the pre-existing rabbit populations within the grids were not quantified and, hence, we cannot state that the warrens contributed to an increase in the rabbit abundance. Regarding the habitat, our results reveal that warrens should be situated in grids with food coverage of less than 50%, while the use of each individual refuge is greater where food availability in the immediate surroundings is at least 20% and shelter at least 50%. The tube warrens showed significantly greater rabbit use than the other types while there was little difference between the stone and pallet warrens in terms of use.     

Seminal ribonuclease of bulls - biosynthesis in normal and orchitic bulls and correlation with blood plasma hormone levels

In sexually mature and healthy bulls, seminal ribonuclease (AS RNase) is synthesized in the distal part of the corpus epididymidis, the cauda epididymidis, the ampullary glands and the seminal vesicles. Indirect immunofluorescence demonstrated AS RNase binding to the cytoplasmic droplets of bull spermatozoa.In bulls with orchitis, AS RNase synthesis decreases in accordance with the degree of damage to the Leydig cells and the drop in the blood plasma testosterone level. The organ most sensitive to decreased testosterone levels, from the aspect of AS RNase synthesis, is the epididymis and the least sensitive are the seminal vesicles. Hypertrophy of the adrenal cortex (in particular of the zona fasciculata and the zona reticularis) and elevated adrenocortical secretion - demonstrated by a raised cortisol concentration in the blood plasma of severely orchitic bulls - failed to inhibit AS RNase synthesis. Injections of Gn RH-LH and HCG raised the blood plasma cortisol, but not testosterone, concentrations in bulls with very severe orchitis. This also indicates serious damage to the Leydig cells in these bulls.     

Non-breeding feather concentrations of testosterone, corticosterone and cortisol are associated with subsequent survival in wild house sparrows

Potential mechanistic mediators of Darwinian fitness, such as stress hormones or sex hormones, have been the focus of many studies. An inverse relationship between fitness and stress or sex hormone concentrations has been widely assumed, although empirical evidence is scarce. Feathers gradually accumulate hormones during their growth and provide a novel way to measure hormone concentrations integrated over time. Using liquid chromatography-tandem mass spectrometry, we measured testosterone, corticosterone and cortisol in the feathers of house sparrows (Passer domesticus) in a wild population which is the subject of a long-term study. Although corticosterone is considered the dominant avian glucocorticoid, we unambiguously identified cortisol in feathers. In addition, we found that feathers grown during the post-nuptial moult in autumn contained testosterone, corticosterone and cortisol levels that were significantly higher in birds that subsequently died over the following winter than in birds that survived. Thus, feather steroids are candidate prospective biomarkers to predict the future survival of individuals in the wild.     

The simulation effects of mountain climbing training on selected endocrine responses

The simulation effects of mountain climbing exercise training on plasma testosterone, cortisol and luteinizing hormone (LH) levels were examined in ten recreational mountain male climbers. Subjects underwent a simulating mountain climbing exercise training 3 times a week for a total of eight weeks before an expedition to Mount Muztag Ata (7546 m, Xingian, China). During training, each subject carried a 40 kg back pack while walking on a treadmill at a speed of 1.9 mph for 60 min at sea level. Subjects completed an incremental treadmill test to exhaustion prior to training, after training, and one week after returning from Mount Muztag Ata. Blood samples were collected from antecubital vein at rest and at 5, 60, and 120 min post testing to determine the plasma testosterone, cortisol and LH levels. The basal plasma testosterone and cortisol concentrations were lower in both post-training and after-climbing conditions compared with that in the pre-training condition (p<0.01). The basal plasma LH concentration was remained unchanged after training and after the mountain climbing compared with levels measured in the pre-training phase. No correlation could be established between plasma LH and testosterone level. These results suggest that an eight-week period of mountain climbing training protocol may be beneficial in maintaining normal endocrine function during and after high altitude mountain expedition. Our results also indicate the decrease of plasma testosterone was LH independent.     

Improving predictions of the location and use of warrens in sensitive rabbit populations

The location and use of warrens is a key factor in the population dynamics of the European rabbit Oryctolagus cuniculus L., which can influence stability and persistence in the long term. Within the species' original distribution range, its numbers have declined sharply in recent decades, which is a serious problem for the conservation of Mediterranean ecosystems. Artificial warrens are commonly used to bolster rabbit populations and the characterization of natural warrens in those areas might improve their efficiency. In this study, we use binomial generalized linear models to identify the factors associated with the location and use of warrens in a low-density area and we evaluate the effect of including the spatial structure of the data in the model. In addition, we generate a map that predicts the most suitable areas for artificial warrens. Contrary to what was expected, habitat variables are only secondary factors, and the location and use of warrens is influenced mainly by spatial factors, such as proximity to nearby warrens. Furthermore, the aggregated spatial pattern of warrens suggests that, at the local scale, for example, the hunting estate, intra-specific interactions might be playing a primary role in these low-density populations. To identify the most suitable sites for artificial warrens and, thereby, improve the efficiency of artificial warren building for conservation purposes, information about spatial structures should be included in models predicting natural warrens of European rabbits.     

A preliminary investigation of urinary testosterone and cortisol levels in wild male mountain gorillas

Urinary steroid hormone levels were measured in wild male mountain gorillas (Gorilla gorilla beringei) to determine how levels of testosterone and cortisol corresponded with age and social rank. Urine samples were collected noninvasively from 18 males, ranging in age from 3–26 years, in three groups of wild mountain gorillas at the Karisoke Research Center, Rwanda, Africa, and samples were analyzed using radioimmunoassay procedures. Males were classified as being immature (<7 years), maturing (10–13 years), or adult (+13 years of age). Immature males had significantly lower levels of testosterone and higher levels of cortisol than both maturing and adult males. No differences in testosterone or cortisol levels were found between maturing and adult males. Dominant males exhibited a trend toward significantly higher levels of testosterone than subordinate males, but no difference was found between cortisol levels of dominant and subordinate males. These results suggest that the increase in testosterone associated with puberty occurs prior to any outward sign of development of secondary sexual characteristics. Within-group male–male competition may affect testosterone levels, but the lack of difference in cortisol levels between dominant and subordinate males suggests that subordinate males are not socially stressed, at least as measured by cortisol. Am. J. Primatol. 43:51–64, 1997. © 1997 Wiley-Liss, Inc.     

Cortisol affects testicular development in male common carp, Cyprinus carpio L., but not via an effect on LH secretion

Previous work showed that prolonged elevated cortisol levels, implicated in the stress adaptation, inhibits testicular pubertal development in male common carp, as well as an impairment of the synthesis of the 11-oxygenated androgens. This may be a direct effect of cortisol on the testis or via the gonadotropin secretion by the pituitary. The aim of the present study was to investigate whether cortisol has an effect on pituitary LH secretion. Juvenile common carp were fed with cortisol containing food pellets. Elevated cortisol levels blocked the increase in testosterone levels and pituitary LH content, but induced higher plasma LH levels at the end of puberty. The in vitro LH release capacity was correlated to the pituitary LH content. At the final stage of pubertal development, when a significant difference in pituitary LH content was observed, sGnRHa-induced LH release was also decreased. Testosterone has been shown to induce development of pituitary gonadotrophs, leading to an increase in LH content and GnRH-inducible LH release, but a decrease in plasma LH levels. We observed decreased plasma testosterone levels as a consequence of prolonged cortisol treatment. It is hypothesised that cortisol inhibits the testicular testosterone secretion and thereby, prevents LH storage. In vitro, this leads to a reduced GnRH-inducible LH release, but in vivo to increased LH plasma levels. It is very unlikely that the impaired testicular development is due to an effect of cortisol on LH secretion.     

Cortisol and glucose responses in juvenile brown trout subjected to a fluctuating flow regime in an artificial stream

In an artificial stream channel, wild 1 year old brown trout Salmo trutta were exposed to fluctuations in flow and water level to simulate hydro-peaking conditions downstream of a hydropower installation. Blood plasma cortisol concentrations reached a maximum of 59·4 ± 35·3 ng ml⁻¹ (mean ± 95% CL) 2h after the end of down-ramping. Return to the pre-exposure cortisol level was achieved within 6 h. When subjected to daily cyclical fluctuations over 7 days, plasma cortisol levels were significantly elevated (61·3 ± 26·8 ng ml⁻¹) on the first day compared to undisturbed fish (4·9 ± 3·7 ng ml⁻¹). On the fourth and seventh day, no elevation in plasma cortisol above control levels was observed. No changes in blood glucose that could be attributed to the stressor were found. There was no correlation between plasma cortisol and blood glucose levels. The short-lived cortisol response to daily fluctuations indicates a rapid habituation to this Stressor.