A re-examination of the subspecies of Red deer (Cervus elaphus) with particular reference to the stocks in Britain

In 1906, Lönnberg identified a native subspecies of Red deer Cervus elaphus scoticus L., in Britain after examining a sample of stags' skulls from Glenquoich (Inverness-shire, Scotland). In north-west England, it is commonly believed that "Red deer of indigenous stock have persisted in this area (Furness Fells, Lancashire, England) from time immemorial…" (Chard, 1966). Arrangements were made to introduce animals into a park within this area in 1970, which, in view of this local opinion, would have been the only Red deer within the district not of native origin. Therefore, it was thought desirable to examine all the stocks of wild or feral Red deer in Britain and to determine which, if any, of them could be considered native and thus deserving of special measures to ensure their survival.
After examining samples of skulls, using multivariate analysis, it appeared that, within the limits of the material representing the various subspecies of the world, there was little support for the concept of subspeciation in Red deer. In Great Britain, however, there appeared to be two distinct forms of Red deer leading a free existence, one presumed to be native and the other to be of park derivation. Visually, the differences were indetectable and could be demonstrated only by discriminant analyses.     

Successful in vitro production of embryos in the red deer (Cervus elaphus) and the sika deer (Cervus nippon)

The aim of our study was to define the conditions for IVM and IVF of oocytes in 2 common deer species as models for endangered related subspecies. Immature oocytes were recovered during the breeding season from postmortem ovaries (red deer) or by repeated laparoscopic follicular aspiration (sika deer). Oocytes were cultured for 24 h in IVM medium supplemented with EGF or FSH and follicular fluid. Stag semen was collected by electroejaculation (both species) or by epididymal flushing (red deer) and cryopreserved. For IVF, oocytes were exposed to different concentrations of thawed spermatozoa in a modified Tyrode albumin lactate pyruvate medium supplemented with 20% (v/v) estrus sheep serum for 18 h. After IVF, presumptive zygotes were allowed to develop in vitro for 7 days in synthetic oviduct fluid (SOF) supplemented with fetal calf serum (10%, v/v). In both species, the presence of ovine FSH and follicular fluid improved the in vitro maturation rate. In the sika deer, the optimal sperm concentration for IVF was 10(6)/mL and some fertilized oocytes reached the early morula stage (20 to 25 cells). In the red deer, after IVF with ejaculated or epididymal spermatozoa (2.0 x 10(6)/mL), 20% of zygotes developed to the blastocyst stage (50 to 80 cells).     

Genetic structure of,and hybridisation between,red (Cervus elaphus) and sika (Cervus nippon) deer in Ireland

This study investigated the levels of genetic diversity and variation exhibited by red and sika deer in Ireland, along with the extent and regional location of hybridisation between these two species. Bi-parental (microsatellites) and maternally-inherited (mitochondrial DNA) genetic markers were utilised that allowed comparisons between 85 red deer from six localities and 47 sika deer from 3 localities in Ireland. Population genetic structure was assessed using Bayesian analysis, indicating the existence of two genetic clusters in sika deer and three clusters in red deer. Levels of genetic diversity were low in both red and sika deer. These genetic data presented herein indicate a recent introduction of sika deer and subsequent translocations in agreement with historical data. The origins of the current red deer populations found in Ireland, based on genetic data presented in this study, still remain obscure. All hybrid deer (red/sika) found in this study were found in Wicklow, Galway and Mayo where the ‘red-like’ deer exhibited sika deer alleles/haplotypes, and vice versa in the case of Wicklow. Molecular methods proved invaluable in the identification of the hybrid deer because identification of hybrids based on phenotypic external appearances (pelage and body proportions) can be misleading. Areas where red and sika deer are sympatric need to be assessed for the level and extent of hybridisation occurring and thus need to be managed in order to protect the genetic integrity of ‘pure’ red deer populations.     

Flow cytometric sexing of X- and Y-chromosome-bearing sperm in Sika deer (Cervus nippon)

The objectives of the study were to determine a practical method of using predetermined sexed semen in Sika deer (Cervus nippon). Semen was collected by electro-ejaculation from two Sika stags and transported to the laboratory and separated into X- and Y-chromosome-bearing sperm after analysis and re-analysis (using a modified high-speed cell sorter), or control (unsorted) semen. Eighty-four Sika hinds were inseminated with 2.8 × 10⁷ unsorted (control) or 2.3 × 10⁶ sorted (X or Y) frozen-thawed semen via intra-uterine laparoscopy 58–66 h after removal of intra-vaginal progesterone-impregnated CIDR devices and the administration of 330 IU PMSG at the time of CIDR removal. No significant differences in the post-thaw motility of control (43.4 ± 4.4%), X- (45.3 ± 4.5%) and Y-sorted (43.5 ± 3.2%) samples were recorded. The sorted frozen-thawed sperm (X, 72.5 ± 6.4%: Y, 75.2 ± 5.5%) recorded significantly (P < 0.05) more intact acrosomes following thawing than the unsorted frozen-thawed (68.2 ± 10.2%) sperm. The individual Sika stags had no effect on the post-thaw sperm motility. Sorted frozen-thawed sperm demonstrated a significantly shorter survival time after thawing than the control sperm (P < 0.05). The number of Sika hinds pregnant following insemination with unsorted or control thawed sperm was significantly higher (33/42; 78.6%) than for hinds inseminated with either X- (5/11; 45.5%) or Y-sorted sperm (15/31; 48.4%). Ultimately 14 out of the 15 calves produced by Sika hinds inseminated with Y-sorted sperm were male (92.9%) and 5/5 calves (100%) from Sika hinds inseminated with X-sorted sperm were female. The sex ratio of the calves born to hinds inseminated with sex-sorted sperm significantly (P < 0.05) deviated for the 48.5% (female, 16/33) and 51.5% (male, 17/33) in the control group. All calves were born between 230 d and 243 d of gestation. Male and female calves in the control group had similar birth and weaning weights as calves from hinds inseminated with X- or Y-sorted sperm. In conclusion it can be said that normal calves of the predicted sex may be produced after intra-uterine insemination conducted by laparoscopy with low numbers of sex-sorted cryopreserved Sika sperm.     

Seasonal coat changes in grazing Red deer (Cervus elaphus)

Seasonal change in the coat of four male and four female Red deer (Cervus elaphus) was studied in coat and skin samples taken monthly for two years. Changes in coat length were measured, the times of moult recorded, and the cycle of follicle activity determined from histological preparations.     

Superovulation and embryo recovery in Red deer (Cervus elaphus ) hinds

In two experiments, Red deer hinds were synchronized with intravaginal progesterone and were given 4 d of treatment (3 d before progesterone withdrawal and 1 d after) with an ovine follicle stimulating hormone (FSH) preparation which had a claimed low level of luteinizing hormone (LH) contamination. In Experiment 1, 12 hinds received one of four FSH levels by osmotic minipump. Hinds were run with fertile stags, and laparotomy and embryo recovery were performed 9 d after progesterone withdrawal. The ovulation rates (mean of three hinds per dosage) were 1.0, 2.0, 4.3 and 15.3 (number of corpora lutea counted) for estimated daily dosages rates of 0.036, 0.071, 0.11 and 0.14 units FSH preparation/day; the response to the increasing dosage was exponential (P<0.01). The recovery rate of ova on flushing was 38% (24 63 ), with all recovered ova being fertilized and of transferable quality. In Experiment 2, performed later in the breeding season, eight hinds received 0.14 units FSH/day either by minipump or by intramuscular injection. The mean ovulation rates were 3.0 and 11.0 (a significant difference, P<0.01), respectively, with a recovery rate of 72% (34 47 ), and with only 18 34 ova considered to be of transferable quality. The recovery rate in Experiment 2 was significantly higher than that in Experiment 1 (P<0.001). Overall, the results were better than those previously recorded for red deer, perhaps a function of both the FSH preparation used and an improved progesterone profile in estrus synchronization.     

Sika deer (Cervus nippon) velvet antler extract attenuates prostate cancer in xenograft model

The present study determines whether antler extract (AE) possesses inhibitory effects in a prostate cancer (PC) xenograft model and explores the underlying mechanism. After therapeutic intervention for two weeks, AE significantly inhibited prostate cancer xenograft tumor growth by 65.08%, and prostate-specific antigen (PSA) and serum dihydrotestosterone (DHT) levels. However, AE increased the serum testosterone level compared to the vehicle control group. Furthermore, our investigation of the inhibitory effects on angiogenesis and epithelial-to-mesenchymal transition (EMT)-related genes revealed that AE downregulated matrix metalloproteinase 2 (MMP)-2, (MMP)-9, vascular endothelial growth factor (VEGF), zinc finger protein (SNAIL1), twist-related protein 1 (TWIST1), and zinc-finger E-box-binding homeobox 1 (ZEB1) in vivo. In contrast, AE increased tissue inhibitor of MMP (TIMP)-1, (TIMP)-2, and E-cadherin. The results suggest that AE possesses potent anti-PC activity, and this is the first report on the anti-PC effect of AE in vivo.     

Calving times of Red deer (Cervus elaphus) on Rhum

The distribution of calving times in a population of Red deer on the Isle of Rhum, Inner Hebrides, between 1971 and 1976 is described. Two-thirds of calves were born during a three-week period beginning in the last week of May, though calving extended from early May to late August. Except in 1976, when calving occurred late, the timing of births was similar in all years. Variation in calving time within years resulted from (i) individual differences in calving time, (ii) late conception in hinds which failed to conceive at first oestrus, and (iii) the effect of previous reproductive history: on average, hinds calved one week later after rearing a calf in the previous winter. Gestation lengths of wild hinds were longer than in a captive group. This may have been due to overestimation of a proportion of gestation lengths in wild hinds, but supplementary feeding could also have shortened gestation length in captive animals.     

Annual foods of the Red deer (Cervus elaphus) and the Roe deer (Capreolus capreolus) in the east of England

There have been few studies on the foods of Red deer (Cervus elaphus L.) and Roe deer (Capreolus capreolus L.) in British forests. Notable exceptions are the work of Prior (1968), Henry (1978) and Hosey (1981) on Roe deer. British work on the food of Red deer seems to have been confined to studies of animals on upland swards (Charles, McCowan & East, 1977; Staines ∓ Crisp, 1978). However, a more abundant supply of data on the food and feeding of both Red and Roe deer comes from the Continent (Dragoev, 1964; Gabuzov, 1960; Kolev, 1966; Novikov & Timofeeva, 1964; Siuda, Zorowski & Suida, 1969; Dzieciolowski, 1967 a , b , 1969). Even so the data available were obtained from studies made on individual species of deer. This paper reports the results of an investigation into seasonal changes in the diet of Red and Roe deer coexisting in a conifer forest in the east of England.     

Assessment LOPU-IVF in Japanese sika deer (Cervus nippon nippon) and application to Vietnamese sika deer (Cervus nippon pseudaxis) a related subspecies threatened with extinction

In mammals, recovery of oocytes by laparoscopic ovum pick-up (LOPU) coupled with in vitro production (IVP) of embryos represents a promising strategy for both amplification and genetic management of sparse animals from captive endangered wild species. As integrated technique developed mainly for domestic livestock, LOPU-IVP requires several studies to set up protocols for follicular stimulation or optimization of IVP before envisaging successful transposition to wild species. In deer, many endangered subspecies would be potentially concerned by applying such an approach using common subspecies for protocols optimization. The aim of the present study was to assess efficiency of follicle stimulation using ovine FSH (oFSH) for recovery of oocytes by LOPU in common sika deer (Cervus nippon nippon) before transposition of an optimized methodology for IVP of embryos from endangered Vietnamese sika deer hinds (Cervus nippon pseudaxis). In common sika deer, two doses of oFSH (0.25 and 0.5 U) and two frequencies of administration (12 and 24 h) were compared by monitoring of subsequent ovarian response, quality of oocytes recovered by LOPU, and in vitro developmental competence. In a first experiment, the dose of oFSH administered did not significantly affect the total number of follicles aspirated per hind per session (8.6 ± 1.0 vs. 8.2 ± 1.6 with 0.5 vs. 0.25 U oFSH, respectively; not significant). In a second experiment, frequency of 0.25 U oFSH administration did not affect ovarian response. Efficiency of IVP determined on blastocysts rates after in vitro maturation, fertilization, and development in oviduct epithelial cells coculture was increased when FSH was administered at 12-h intervals. Immune response after several follicular stimulations was detected against exogenous oFSH in plasma from the majority of sika deer hinds but was not associated with decreased ovarian response. When 0.25 U oFSH was administered at 12-h intervals to Vietnamese sika deer (N = 4), good quality cumulus oocyte complexes with complete and compact cumulus investments were recovered allowing a high cleavage rate after in vitro maturation and fertilization. Development to the blastocyst stage occurred in a high proportion (30% of oocytes) after coculture with ovine epithelial cells allowing cryobanking of transferable embryos from Vietnamese sika deer. These results confirm that LOPU-IVF after ovarian stimulation with oFSH may be a successful tool for cryobanking transferable embryos from endangered sika deer subspecies.     

Teeth as indicators of age with special reference to Red deer (Cervus elaphus) of known age from Rhum

An experiment in land management and Red deer population control was begun by the Nature Conservancy on the Isle of Rhum, a National Nature Reserve off the west coast of Scotland, in 1957. To assess the results fully it has become necessary to determine the most suitable ageing method which can be applied to the material, which has accumulated since from the annual deer culls and natural deaths.
Using mandibles only from individuals, whose age was certainly known by marking at birth, and which, in most cases were shot by mistake during subsequent annual culling operations, four different techniques were critically tested. These comprised correlating various combinations of jaw and tooth measurements with age, examining the nature and quantity of secondary dentine deposited within the crown of the first incisor, interpreting the number of layers in the pad of cementum beneath the crown of the first molar, and assessing the stages reached in tooth replacement, eruption and wear. Of these various methods, ageing by tooth replacement, eruption and wear proved the most reliable.     

Bovine microsatellite loci are highly conserved in red deer (Cervus elaphus), sika deer (Cervus nippon) and Soay sheep (Ovis aries)

We tested 174 bovine microsatellite primer pairs for use in a primitive breed of sheep and two species of deer. Of 173 markers, 127 (73·4%) gave a product in Soay sheep ( Ovis aries ) of which 54 (42·5%) were polymorphic. One hundred and twenty-nine of 174 (74·1%) markers gave a product in red deer ( Cervus elaphus ) of which 72 (55·8%) were polymorphic. In sika deer ( Cervus nippon ) 126 of 171 (73·7%) microsatellite primers gave a product with 47 (37·3%) polymorphic. The proportion of bovine microsatellite loci conserved across artiodactyl species was significantly greater in this study than previously reported. Reasons for this high degree of microsatellite conservation are discussed. We suggest that a high resolution comparative map of the artiodactyls can be constructed using microsatellites.     

The timing of reproduction in Red deer (Cervus elaphus) in relation to latitude

A survey was carried out of the birth dates of Red deer calves born in zoological gardens throughout the world in an attempt to assess the effects of latitude on the timing of conception. The results showed that Red deer in the Northern Hemisphere calve at a similar time of the year irrespective of latitude, and that there is no increase in the length of the calving season as the equator is approached; south of the equator, calving is shifted by six months. Populations living at similar latitudes can show small variations of up to two weeks between the median calving dates.     

四川梅花鹿春季昼夜活动节律与时间分配

1997年4月,在四川铁布自然保护区用直接观察法对220头次四川梅花鹿群的昼夜活动节律和时间分配进行了观察。结果表明,鹿群春季昼夜活动的规律性较强。白昼,鹿群活动呈现明显的双峰型,2个高峰时段为08：30和19：30前后,活动频率分别为71．9％和94．13％,其他时间多处于休息状态;鹿群夜间活动强于昼间,活动频率均在6l％以上,仅在半夜01：30前后有一个相对不活跃期。同时发现,春季活动高峰期约90％的活动时间被四川梅花鹿用于采食和移动。这可能与光照、食物及人类活动的影响有关。     

Variation in mandible length and body weight of Red deer (Cervus elaphus)

The range of intra-specific variation in morphological characters has rarely been examined by taxonomists, and subspecies have often been named on the basis of as few as two skulls. However, despite the limitations set by the material data, taxonomists have long recognized an apparent cline in nose length of Red deer ( Cervus elaphus L.) between the eastern and western extremities of its European distribution. Since larger samples of mandible and body weight data have recently become available for three widely separated populations within the north western parts of their range—the Isle of Rhum (north-west Scotland), Grizedale (north-west England) and Oksbol (Denmark)—it has become possible to examine this cline critically in relation to age, development and geography. In all three populations, the relationship between body weight and jaw length differed significantly at the 1% level, but, as expected, whilst the deer from Denmark had the longest mandibles as adults, as juveniles they had the shortest mandibles in relation to body weight. Furthermore, if the island of Rhum was divided arbitrarily into three areas, namely north western, south western and eastern, an exactly similar picture emerged. Therefore, no single simple law such as Bergman's rule seems to offer an entirely satisfactory explanation.     

Variable extent of hybridization between invasive sika (Cervus nippon) and native red deer (C. elaphus) in a small geographical area

Instances of hybridization between endemic and alien species pose a threat to species integrity but also provide us with an opportunity to study the dynamics of gene flow between two species as they first meet. Here, we used variation at 22 highly differentiated microsatellite loci and one mitochondrial DNA (mtDNA) marker in a sample of 735 individuals, to investigate the genetic consequences of an introduction of Japanese sika deer ( Cervus nippon ) for native red deer ( C. elaphus ) on the Kintyre Peninsula in Scotland. We investigated population structure, estimated null-allele frequency and assigned individual hybrid scores using a Bayesian clustering algorithm implemented in structure 2.2. The dataset clearly divided into two clusters and generally, introgression into red and sika was low. However at one site, West Loch Awe, 43% of individuals were hybrids. MtDNA introgression indicated that hybridization was occurring between red-deer hinds and sika-deer stags. We argue that the pattern of differential introgression across the study area is primarily due to the rarity of hybridization events between the two species and the limited time the two species have been in contact (< 120 years). This contrasts with the causes of classic mosaic hybrid zones (selection induced by habitat variability). Currently, it seems possible that, in time, the level of hybridization found at West Loch Awe could also be found across the whole of the peninsula.