The effect of artificial photoperiodicity and antiandrogen treatment on the antler growth and plasma levels of LH, FSH, testosterone, prolactin and alkaline phosphatase in the male white-tailed deer

1. Artificial extension of day-length in adult male white-tailed deer during the autumn induced: (a) premature casting of antlers, early onset of the new antler growth and out of season mineralization, (b) early elevation of plasma levels of prolactin, LH, FSH, testosterone and alkaline phosphatase and (c) out of season hair molt. 2. Intramuscular administration of the antiandrogen cyproterone acetate immediately after velvet shedding induced: (a) dramatic reduction of testosterone levels in plasma, (b) premature casting in bucks with fully mineralized antlers and (c) renewal of bone rebuilding activity in incompletely mineralized antlers which resulted in blockage of casting.     

Influence of plane of winter nutrition on plasma concentrations of prolactin and testosterone and their association with voluntary food intake in red deer stags (Cervus elaphus)

Blood samples were taken at weekly or 3-weekly intervals from 12 red deer stags, from 3 weeks of age until 3 years of age. Six of the stags were fed to appetite throughout the study and six were maintained on 70% as much during each winter and to appetite each summer. All plasma samples were analysed for testosterone and prolactin. Food intake was measured daily. Annual rhythms of testosterone, prolactin and food intake were evident. Peaks of prolactin, on an increasing photoperiod tended to occur slightly before peaks of food intake and troughs of food intake on a decreasing photoperiod occurred at or slightly after peaks of testosterone. The reduced plane of winter nutrition tended to delay rather than alter the amplitude of the annual cycles of testosterone, prolactin or food intake. Several of the unrestricted stags showed biannual rhythms of testosterone, prolactin and food intake accompanied by additional cycles of coat development and antler growth. These additional cycles tended to be 6 months out of rhythm with normal cycles. It is considered that this is due to the stags being stimulated by a particular day-length irrespective of whether it is decreasing or increasing and that plane of nutrition may influence the control of phase blockers.     

Requirement of thyroid function for the expression of seasonal reproductive and related changes in red deer (Cervus elaphus) stags.

The effect of thyroid function on regulation of seasonal reproduction was investigated in three red deer stags thyroidectomized (THX) in summer (January 1988) in comparison with five thyroid-intact controls. Responses of luteinizing hormone (LH) and testosterone to a bolus injection of 10 micrograms gonadotrophin-releasing hormone (GnRH) were tested in July, October, December, February and April. Blood samples were collected at weekly intervals from December 1987 to June 1989 for measurement of testosterone, triiodothyronine (T3) and prolactin concentrations. Testis diameters were measured every 2 weeks. In October 1988 (spring), plasma LH concentrations of control stags were less responsive (P less than 0.01) to stimulation by GnRH than those of THX stags; plasma testosterone concentrations and testis diameters were low and there was no increase in plasma testosterone concentrations after injection of GnRH in control stags during October or December (spring, early-summer). In contrast, THX stags maintained a testosterone response (P less than 0.01) in these 2 months and did not exhibit any signs of a seasonal lack of reproductive activity at this time of year. Control stags cast antlers in spring whereas THX stags maintained hard antlers throughout the study. Concentrations of plasma T3 were not detected in THX stags from June 1988 onwards, but exhibited a seasonal pattern in control stags, with low concentrations during autumn and winter (April to July) and high concentrations in spring and summer (August to February). There was no effect of thyroidectomy on the seasonal pattern of prolactin secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

The induction of male sexual behavior in red deer (Cervus elaphus) by the administration of testosterone to hinds and estradiol-17β to stags

One-gram implants of testosterone were placed subcutaneously in two adult intact and one castrated red deer hinds, and 100-mg implants of estradiol-17β were given to three castrated and two intact red deer stags. Their sexual behavior was then observed in the wild and in captivity for up to 3 years. Testosterone initially produced prolonged and intense estrous behavior in the hinds; this gradually gave way to behavior normally shown only by rutting stags, such as flehmen, herding threats, and roaring. Testosterone-implanted hinds rose in the dominance hierarchy as measured by competition for food. Pregnancy, or the administration of progestagens, suppressed most of this testosterone-induced behavior. Testosterone also induced development of male secondary sexual characteristics such as formation of small antler pedicles (but not antlers), hypertrophy of the clitoris and neck musculature, and development of a neck mane and male rutting odor. Estradiol-17β mimicked the behavioral effects of testosterone when given to castrated stags, stimulating all components of rutting behavior. In intact stags, the only effects were to abolish antler casting and stimulate roaring; normal rutting behavior continued unchanged. In contrast to the effects of testosterone, estradiol-17β did not influence the social status of either intact or castrated stags. The distinctions between the behavioral effects of androgens and estrogens are ill defined and are determined more by the prior sexual differentiation of the brain and the duration of steroid treatment, than by the nature of the steroid.     

Effects of environmental conditions, human activity, reproduction, antler cycle and grouping on fecal glucocorticoids of free-ranging Pampas deer stags (Ozotoceros bezoarticus bezoarticus)

In this study, a commercial enzyme immunoassay (EIA) was validated in detecting glucocorticoids in Pampas deer feces, in order to investigate the influence of several factors on the adrenocortical function. Fecal samples, behavioral data and information concerning male grouping and antlers status were collected at a monthly basis during a 1 year period from free-ranging stags living at Emas National Park, Brazil (18 degrees S/52 degrees W). The results revealed that concentrations of fecal glucocorticoids in winter were significantly higher than those corresponding to spring and summer. In addition, dry season data presented higher levels than during the wet season. Significant difference was found between fecal levels of breeding stags in summer and nonbreeding stags, whereas no difference was observed between breeding stags in winter and nonbreeding stags. On the other hand, males from areas with frequent human disturbance exhibited higher glucocorticoid concentrations and flight distances than individuals from areas of lower human activity. Males with antlers in velvet had elevated levels compared with animals in hard antler or antler casting. Also, we found that glucocorticoid levels were higher in groups with three or more males than in groups with only one male. The flight distances showed positive correlation with fecal glucocorticoid. These data indicate that fecal glucocorticoid provides a useful approach in the evaluation of physiological effects of environment, inter-individuals relationship and human-induced stressors on free-ranging Pampas deer stags.     

Effect of antiandrogen cyproterone acetate on the development of the antler cycle in Southern pudu (Pudu puda)

The antler cycle of pudu is similar to other cervids, but unlike most boreal deer, male Southern pudu (Pudu puda) exhibits two seasonal peaks of LH and testosterone. In that respect, pudu is similar to roe deer. Whereas the antler cycle in some deer species, such as roe deer or white-tailed deer, is very sensitive to variation of testosterone, in other cervids, such as fallow deer or reindeer, a blockade of androgens with cyproterone acetate (CA) has little or no effect on the timing of the antler casting. In order to test the sensitivity of pudu antlers to variations of androgens, CA (administered 2x weekly at 50 mg/buck) was injected intramuscularly for 3 weeks in 5 adult male pudu, starting February 19 (late summer). Four other males of similar age served as controls. The experiment was performed at the University of Concepcion, Chile, latitude 36.6 degreeS. Blood samples were taken once a week between January 19 and April 3. In CA-treated bucks, the antlers were cast approximately 3 weeks after the initiation of CA treatment and a new antler growth began almost immediately. The antlers reached about 5 cm in length, before ceasing to grow at the end of April, when they became mineralized and were subsequently polished. CA had no effect on the already declining levels of LH. Plasma levels of testosterone in controls increased from February 15, whereas in CA-treated bucks remained depresses until March 21. It is concluded that similarly to white-tailed deer, the antler cycle of Southern pudu is very sensitive to manipulation of androgen levels.     

Regrowth of amputated velvet antlers with and without innervation

The influence of removing portions of the growing antler of yearling red deer stags on subsequent regeneration of the antler in the same season was studied. The influence of the innervation of the antler on such regeneration was the subject of a further study. When the top 0.5-1 cm was removed from antlers 9-17 cm long, growth was slightly reduced in that season. When the antler/pedicle length was reduced to 6-10 cm in antlers 16-38 cm long, branched antlers regrew in 11 out of 13 cases provided the amputation was carried out early in the growing season, i.e., before mid-December. Denervated antlers were shorter, lighter, and of different shape compared with controls, but they were of similar density. Denervation was confirmed histologically. Cleaning of velvet and casting of antlers following castration were unaffected by denervation. It would appear that although nerves affect the size and shape of the antler, they are not essential to the actual control of antler growth and regeneration.     

Influence of photoperiod on the seasonal pattern of secretion of luteinizing hormone and testosterone and on the antler cycle in roe deer (Capreolus capreolus).

Annual variations in concentrations of luteinizing hormone (LH) and testosterone in plasma were analysed in relation to the antler cycle in six adult male roe deer exposed to a natural photoperiod (latitude 46 degrees 10'N) and in four adult males maintained in a constant short-day photoperiod (8 h light: 16 h dark) for a year, from the winter solstice at which time both groups of animals had antlers in velvet. The animals were sampled, every 15 min for 2 or 4 h at intervals of one month for a year. Under both natural and experimental conditions, LH concentrations were high from January to March, but in the experimental conditions they decreased between April and May-June, whereas in the natural conditions they increased. Plasma LH concentration was lowest between July and November in animals under natural photoperiod, whereas under 8 h light:16 h dark photoperiod a second increase in plasma LH occurred between August and September. Between March and August, concentrations of plasma testosterone increased under natural photoperiod, whereas under experimental photoperiod there was a biphasic pattern of plasma testosterone with peaks between February and May and between September and November. Under natural photoperiod, antlers were cast in November, 369 +/- 6 days after the previous antlers were cast. Under experimental photoperiod, antlers were cast after 193 +/- 10 days, and a new set developed. The sexual cycle of the male appears to be initiated by an endogenous rhythm in winter and is then maintained by hormonal changes resulting from increasing photoperiod in spring.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evidence that the seasonal antler cycle of adult Reeves' muntjac (Muntiacus reevesi) is not associated with reproductive quiescence

Data were obtained from post-mortem investigations of 190 culled and road-killed muntjac bucks between 1967 and 1989. Although adult bucks (i.e. those having undergone at least one antler cycle) have a synchronous annual antler cycle, unlike temperate-zone cervids there was little seasonal variation in testis size or activity, or in the size and activity of the epididymidis or accessory reproductive glands. Spermatogenesis was not abated when the antlers were in velvet and year-round fertility was achieved without additional sperm storage. There was little seasonal change in plasma testosterone concentrations in samples obtained from captive and free-living bucks although castration caused antler casting and prevented mineralization. Hence the data are equivocal as to the role of steroids in driving the antler cycle; experimental work on this species would be valuable in examining the mechanisms which regulate the antler cycle.     

Long-term stimulatory effects of a continuous infusion of LHRH agonist on testicular function in male red deer (Cervus elaphus)

Red deer stags were infused continuously with the LHRH agonist buserelin at 180-270 micrograms/day (1.2-1.8 micrograms/kg/day) for 72 days starting in late winter with the aim of suppressing reproductive function and inducing premature casting of the antlers. Contrary to expectation, the treatment resulted in a long-term stimulation of testicular activity lasting at least 2 months; the increases in plasma concentrations of testosterone were associated with an increase in aggressive behaviour and the development of rutting odour in the urine. The stags cast their antlers at the normal time in spring after the end of the treatments. The results indicate that the pituitary gonadotroph cells in the stag can continue to secrete LH in response to chronic exposure to an LHRH agonist and do not become rapidly desensitized. The effect of the agonist is therefore to cause significant stimulation of testicular activity which is a conspicuous response in the non-breeding season when the stags are already in a hypogonadal state.     

Seasonal changes in fecal testosterone concentrations and their relationship to the reproductive behavior, antler cycle and grouping patterns in free-ranging male Pampas deer (Ozotoceros bezoarticus bezoarticus)

The purpose of this study was to validate noninvasive endocrine monitoring techniques for Pampas deer and to evaluate seasonal changes in testicular steroidogenic activity and their correlation to reproductive behavior, antler cycle and group size. Thus, fecal samples, behavioral data and observations of antler status were collected at monthly intervals during 1 year from free-ranging Pampas deer stags (three radio-collared individuals and 15 random individuals) living in Emas National Park, Brazil (18 degrees S latitude). Fecal steroids were extracted using 80% methanol and steroid concentrations were quantified by a commercial enzyme immunoassay (EIA). Fecal testosterone concentrations peaked in December-January (summer), March (early autumn) and in August-September (winter-spring), with minimal values from April-July. Reproductive behavior had two peaks, the first in December-January, characterized by predominately anogenital sniffing, flehmen, urine sniffing, chasing and mounting behavior, and the second peak in July-September (behavior primarily related to gland marking). There were significant correlations between fecal testosterone and reproductive behavior (r=0.490), and between fecal testosterone and antler phases (r=0.239). Antler casting and regrowth occurred under low testosterone concentrations, whereas velvet shedding was associated with high concentrations of testosterone. We inferred that Pampas deer stags exhibited a seasonal cycle that modulated sexual behavior and the antler cycle, and we concluded that fecal steroid analysis was a practical and reliable non-invasive method for the evaluation of the endocrine status of free-ranging Pampas deer.     

Pedicle and antler fusion and double-head formation in a sika stag (<Emphasis Type="Italic">Cervus nippon</Emphasis>)

A case of pedicle and antler abnormality in a 12–14 year old sika stag shot in January 2003 in Schleswig-Holstein (Germany) is presented. The abnormality combines fusion of pedicles and antler bases with subsequent double-head formation. Double-heads result from growth of new antler bone without casting of the previous hard antlers. In consequence, two consecutively formed antlers are present in an individual. The stags skull showed a plate-like osseous structure whose broad and slightly elevated central portion was identified as the fused pedicles. The peripheral parts of this osseous plate constituted the second antler growth of the (former) double-head. A cast pair of antlers, which were fused at their bases, had been found in a neighbouring hunting district in the summer of 2002. The close fit between the casting surface of the fused antlers and the surface of the pedicle/antler structure on the stags skull indicated that the antlers had been grown by this stag and were belatedly cast from his fused pedicles. The fused antlers thus constituted the first antler growth of the double-head. We suppose that the broad connection between the fused antlers and the fused pedicles prevented antler casting at the normal time and thereby caused the double-head condition. The presentation of this antler abnormality is taken as an occasion to discuss the significance of pedicles for the normal casting and regeneration of antlers.     

Testosterone and estradiol concentrations in serum, velvet skin, and growing antler bone of male white-tailed deer

The growth and mineralization of antlers correlate with the seasonal variation of serum androgens. Whereas seasonal levels of testosterone (T) in plasma are well established, steroid concentrations have not yet been determined in the tissues of growing antlers. Therefore, RIA was used to determine T and 17beta estradiol (E2) in serum, and three areas (tip, middle, and base) of the antler bone and the antler skin, called velvet. Blood and antler tissues of white-tailed deer (Odocoileus virginianus) were collected from May to August. The difference between levels of T and E2 among the sites was calculated using the square root transformation followed by a mixed model analysis with individual deer and an interaction of individual and year (individual(*)year) as a random factor. Concentrations of T in serum (799+/-82 pg/ml) were higher than T values in the velvet (589+/-58 pg/ml, P<0.01) and in the antler bone (538+/-58 pg/ml, P<0.001). Estradiol concentrations differed among antler tissues and serum (P<0.001) and between years (P<0.01). Estradiol concentrations in serum (25+/-25 pg/ml) were consistently lower than those in antler bone (208+/-11 pg/ml, P<0.001) and velvet (150+/-12 pg/ml, P<0.001). The E2:T ratio in serum was 1:10-60. The same ratio for the antler bone was only 1:2-3 and for the velvet 1:3.5. It is concluded that higher T and lower E2 concentrations found in plasma, as compared to antler bone or antler velvet, may indicate a partial metabolism of systemic androgens into estrogens xin the tissues of growing antlers.     

LH and testosterone responses to GnRH in red deer (Cervus elaphus) stags kept in a manipulated photoperiod

Six red deer stags from age 4 months were kept in a light-proof room under an artificial photoperiod consisting of 5.5 cycles of alternate 2-month periods of 16 h light and 8 h dark (16L:8D) and 8L:16D. At 2 or 3 weekly intervals from 10 months of age through 4 cycles, the stags were anaesthetized with xylazine and challenged i.v. with 10 micrograms GnRH. Blood samples were withdrawn immediately before and 10 and 60 min after injection. LH and testosterone concentrations were measured in all samples by RIA. Antler status was recorded daily. Peak LH values on each sampling day occurred in the sample taken 10 min after GnRH stimulation while peak testosterone occurred in the sample taken at 60 min. There were 4 cycles of LH and testosterone secretion accompanied by 4 antler cycles in the stags. The highest LH responses were detected during short days (8L:16D), and the highest testosterone responses were detected around the time of the change from short to long days. The responses of both hormones were lowest at the end of periods of long days or the beginning of short days. The increased pituitary LH response to GnRH was evident 4 weeks after the change to short days which are stimulatory for gonadal development. Antler casting occurred at the end of long days and cleaning at the end of short days. It is considered that antler cycles were due to the ability of the stags to vary release of LH and testosterone in response to changes in the artificial photoperiod.     

Territory Establishment and Antler Cycle in Male Roe Deer

Territory establishment and antler cycle in male roe deer, Capreolus capreolus, were studied in two areas in south-eastern Sweden during 1989–1993. Initial territorial behaviour was observed in early Mar., when overlap between male ranges decreased from 66 % to 0 % over a 3–6-wk period, as determined from telemetry data. The decrease was associated with an increased number of agonistic interactions. Most territories were established at the end of Mar. to early Apr., with a tendency for males re-establishing old territories to be first. In all years, territories were taken up 1–3 wk before males shed velvet from their antlers. The distance between geometric centres of neighbouring males in winter and after territory establishment increased from 164 to 364 m. Former territory holders moved the geometric centres of their winter range and subsequent territory a shorter distance than first-year territorials, 129 and 267 m respectively. Territory defence ceased after the rut in late Aug., and the spatial relations between neighbours reverted to the situation before the territorial season. Neither velvet shedding nor antler casting was correlated with male age (yearlings excluded), which contradicts earlier statements. Neither were there any associations between timing of antler casting and subsequent velvet shedding, nor vice versa. Hence, a male casting antlers early one year derives no advantage by early shedding the next season. Furthermore, to be in hard antlers is not a prerequisite for territory take-up, whereas antlers probably are necessary for maintaining the area throughout the season. Functional aspects on roe buck territoriality are also discussed.     

The effect of cyproterone acetate on the antler cycle in red deer (Cervus elaphus L.)

The aim of the study was to test the effect of antiandrogen, cyproterone acetate (CA) on the antler cycle in the red deer (Cervus elaphus). CA was administered to three adult red deer stags (Edward, Fuks and Gacek) in weekly intervals. Edward and Fuks were given 600 mg + 600 mg of CA, whereas Gacek was given 600 mg + 300 mg. CA was injected during the hard antler phase: in mid-October (Edward), at the end of November (Fuks) and at the end of January (Gacek). CA caused the antler casting 17 to 22 days after the first injection. In all stags, the casting of antlers was followed by a period of intensive growth of new antlers. Edward was given CA at the end of October. This treatment was responsible for occurrence of the two antler cycles in the year of the experiment. When CA was administered during the middle of the hard antler phase an additional short antler cycle occurs followed by new antler growth. CA treatment in the later part of hard antler phase may cause a prolonged antler cycle.     

梅花鹿(Cervus nippon hortulorum)茸角生长发育各阶段血浆睾酮、雌二醇的含量变化

本文用放射免疫测定法对3个不同年龄组的雄性东北梅花鹿茸角生长发育各阶段外周血中的睾酮、雌二醇含量进行了测定。茸期两种激素差异很大,睾酮含量低,雌二醇高,骨化期二者增高很快。对这两种激素含量的变化与茸角发育各阶段的关系进行了讨论。认为只有在二种激素同时增加时,鹿角才能骨化。     

Induction of antler growth in a congenitally polled Scottish red deer stag.

A congenitally polled red deer stag was captured from a Scottish deer forest and kept in an enclosure for observations. The animal had rudimentary antler pedicles but no antlers, and during five years of study no significant antler development occurred. Amputation of the apex of one antler pedicle in May 1974 when the stat was 12 years of age resulted in the growth of a complete antler on the operated side, and this antler was subsequently cleaned and cast in the normal way and a new antler cycle was initiated. The result illustrates that the primary abnormality in this polled stag lay not in his inability to grow antler, but in his inability to develop fully formed antler pedicles from which normal antler tissue could differentiate. Traumatizing the rudimentary pedicle had the effect of stimulating growth of antler tissue, and once this was formed the process of cleaning, casting and regrowth occurred spontaneously. The incomplete development of the antler pedicles is considered to be responsible for the absence of antlers in the majority of "hummels" in Scotland, and the etiology of the condition is discussed.     

The mechanism of antler casting in the fallow deer.

The process by which antlers are detached from their pedicles was examined histologically in fallow deer castrated in the autumn to induce precocious casting. Osteoclastic erosion across an abscission line between the dead bone of the antler and the living bone of the pedicle was found to be responsible for the separation of the 2. As early as 3 days after castration, osteoclasts and associated lacunae were present on the sides of the pedicle bone. These were then found in progressively deeper locations, by 2 weeks extending across the entire width of the pedicle. Concomitant with the centripetal spread of osteoclasts was the enlargement of Haversian canals, the surfaces of which became lined with osteoclasts. These widening vascular channels within the bone were filled with connective tissue, which in precasting stages formed a mesodermal pad about 1 mm thick. In later stages, a circumferential cleft was excavated beneath the antler burr, and connective tissues from the surrounding pedicle skin invaded the space between the antler and pedicle. After casting, the ingrowing integumental tissues fused with the mesodermal tissues derived from the vascular channels of the pedicle to give rise to an incipient antler bud beneath the scab. The ingrowth of epidermis capable of de novo hair follicle formation gave rise to the future velvet skin that envelops the elongating antler.     

Seasonal variations in semen characteristics, semen cryopreservation, estrus synchronization, and successful artificial insemination in the spotted deer (Axis axis)

Ten adult male spotted deer were monitored over a 2-year interval to determine seasonal variations in testicular size, semen characteristics and serum testosterone concentrations, and to determine if there was an association between season and type of antler. Mean (+/-S.E.M.) testicular volume (118.8+/-4.6 cm(3)), serum testosterone concentration (1.2+/-0.1 ng/mL), semen volume (4.1+/-0.6 mL), sperm concentration (338.3+/-24.9 x 10(6) mL(-1)), percentage of morphologically normal sperm (79.1+/-2.8%), and percentage of motile sperm (66.5+/-1.5%) were higher (P<0.05) in hard antler deer (peaked from March to May) than in deer with velvet antlers or in deer in which the antler has been shed. Thus, March-May was considered the physiologic breeding season for these deer; at this time, all stags had hard antlers. Furthermore, a Tris-citrate-based semen extender containing 4% glycerol and 20% egg-yolk was adequate for cryopreservation of semen. Estrus was induced with an implant containing norgestomet, timed transcervical AI was done with fresh semen, and 3 of 10 females were pregnant at 60 days, with fawns born 120 (premature), 240 and 243 days after AI. These results were considered a model for the use of assisted reproductive techniques to conserve other critically endangered deer species of India.