Phylogenetic relationships among european red deer, wapiti, and sika deer inferred from mitochondrial DNA sequences

We determined the mitochondrial cytochrome b gene sequences (1140 bp) of one subspecies of the European red deer (Cervus elaphus in Europe), three subspecies of the wapiti (C. elaphus in Asia and North America), and six subspecies of the sika deer (C. nippon in Japan). Our phylogenetic analysis revealed the monophyly of the European red deer, that of the wapiti, and that of the sika deer. The wapiti, however, was shown to be more closely related to the sika deer than to the European red deer. This is in conflict with traditional morphological results, which suggest a close sister group relationship between the wapiti and the European red deer. The divergence time between the European red deer and the wapiti plus the sika deer was estimated to be approximately 0.80 Ma, and that between the wapiti and the sika deer was estimated to be 0.57 Ma. The sika deer was subdivided into two subspecies groups, and the wapiti was also found to consist of an Asian group and a North American group.     

A mitochondrial control region and cytochrome b phylogeny of sika deer (Cervus nippon) and report of tandem repeats in the control region

Sika deer (Cervus nippon Temminck) are endemic to mainland and insular Asia. Numerous subspecies have been named, but they are not quantitatively well defined. Portions of the mitochondrial cytochrome b gene (450 bp) and control region (512 bp) were sequenced from 28 individuals belonging to five sika subspecies and two Cervus elaphus subspecies. Phylogenetic trees constructed using these sequences clearly demonstrated that sika are monophyletic with respect to C. elaphus. A survey of variation in the control region showed that approximately half the variation occurred in a 100-base segment between positions 150 and 250 in the left domain of the control region. Within this region there were three tandemly repeated copies of a 39-base motif. In addition, two of the samples (C. n. aplodontus and C. n. hortulorum) contained, respectively, two and four additional copies of the repeated motif.     

The establishment of a hybrid zone between red and sika deer (genus Cervus)

Japanese sika deer ( Cervus nippon nippon ) were introduced to Scotland around 80 years (20 generations) ago. The sika phenotype is expanding its range and hybridizing extensively with native red deer ( Cervus elaphus ) leading to the establishment of a hybrid zone. This zone is currently moving and cannot be considered to be at equilibrium. Cervid genotypes and mitochondrial haplotypes were mapped across the sika phenotype range, using diagnostic protein isozymes, microsatellite nuclear DNA markers and RFLPs in mtDNA. These were analysed to estimate heterozygote deficits and nuclear linkage disequilibria and cytonuclear disequilibria in relation to gene frequencies and time since contact. Introgression was found in both taxa and strong linkage disequilibria and heterozygote deficits characterize the populations longest exposed to hybridization. Populations further from the introduction site, where hybridization is facilitated by the dispersal of sika-like stags, show low values for linkage disequilibria and heterozygote deficit. The observed patterns in genotype are explained in terms of assortative mating and a selective advantage of the sika genotype. The genetic integrity of the Scottish mainland red deer is shown to be at risk from the invasion of sika.     

Polymorphism and genetic control of erythrocyte 6-phosphogluconate dehydrogenase in the genus Cervus

A study of 11 enzyme systems in blood samples of Cervus dama, C. elaphus, C. nippon and hybrids C. elaphus X nippon has revealed an erythrocyte 6-phosphogluconate dehydrogenase polymorphism in the hybrid populations. Genetic analysis suggests that this enzyme is controlled by one gene locus with two codominant alleles, one specific for pure Japanese sika deer, the other for pure red deer as well as for fallow deer, while both alleles have been found in the red X sika hybrids.     

Introgression of exotic Cervus (nippon and canadensis) into red deer (Cervus elaphus) populations in Scotland and the English Lake District

Since the mid‐19th century, multiple introductions of Japanese sika deer (Cervus nippon nippon) and North American wapiti (C. canadensis) have taken place in the British Isles. While wapiti have generally been unsuccessful, sika have been very successful, especially in Scotland where they now overlap at least 40% of the range of native red deer (C. elaphus). Hybridization between these two species and red deer has been demonstrated in captivity and in the wild. Using a panel of 22 microsatellite loci that are highly diagnostic between red deer and sika, and moderately diagnostic between red deer and wapiti, we investigated the extent of introgression between these species in 2,943 deer sampled from around Scotland and from the English Lake District using the Bayesian clustering software STRUCTURE. We also used a diagnostic mitochondrial marker for red deer and sika. Our survey extends previous studies indicating little introgression of wapiti nuclear alleles into red deer, in particular in Northern Scotland, Kintyre, and the Lake District. We found a new area of extensive sika introgression in South Kintyre. In the North Highlands, we show for the first time geographically scattered evidence of past hybridization followed by extensive backcrossing, including one red‐like individual with sika introgression, two sika‐like individuals with red deer introgression, and six individuals that were apparently pure sika at the nuclear markers assessed but which carried red deer mitochondria. However, there has not been a collapse of assortative mating in this region. Similarly, in the English Lake District red deer, we found only traces of past sika introgression. No sika alleles were detected in the Central Highlands or the Hebridean red deer refugia. We make suggestions for management to prevent further spread of sika alleles into red deer and vice versa.     

A survey of the hybridisation status of Cervus deer species on the island of Ireland

Red deer (Cervus elaphus) did not recolonise Ireland after the last glaciation, but the population in Co. Kerry is descended from an ancient (c. 5000 BP) introduction and merits conservation. During the mid-19th century exotic species including North American wapiti (C. canadensis) and Japanese sika deer (C. nippon nippon) were introduced to Ireland, mainly via Powerscourt Park, Co. Wicklow. While wapiti failed to establish, sika thrived, dispersed within Co. Wicklow and were translocated to other sites throughout Ireland. Red deer and sika are known to have hybridised in Ireland, particularly in Co. Wicklow, but an extensive survey with a large, highly diagnostic marker panel is required to assess the threat hybridisation potentially poses to the Co. Kerry red deer population. Here, 374 individuals were genotyped at a panel of 22 microsatellites and at a single mtDNA marker that are highly diagnostic for red deer and Japanese sika. The microsatellites are also moderately diagnostic for red deer and wapiti. Wapiti introgression was very low [trace evidence in 2 (0.53 %) individuals]. Despite long-standing sympatry of red deer and sika in the area, no red deer-sika hybrids were detected in Co. Kerry suggesting strong assortative mating by both species in this area. However, 80/197 (41 %) of deer sampled in Co. Wicklow and 7/15 (47 %) of deer sampled in Co. Cork were red-sika hybrids. Given their proximity and that hybrids are less likely to mate assortatively than pure individuals, the Co. Cork hybrids pose a threat to the Co. Kerry red deer.     

Variability of Female Responses to Conspecific vs. Heterospecific Male Mating Calls in Polygynous Deer: An Open Door to Hybridization?

Males of all polygynous deer species (Cervinae) give conspicuous calls during the reproductive season. The extreme interspecific diversity that characterizes these vocalizations suggests that they play a strong role in species discrimination. However, interbreeding between several species of Cervinae indicates permeable interspecific reproductive barriers. This study examines the contribution of vocal behavior to female species discrimination and mating preferences in two closely related polygynous deer species known to hybridize in the wild after introductions. Specifically, we investigate the reaction of estrous female red deer (Cervus elaphus) to playbacks of red deer vs. sika deer (Cervus nippon) male mating calls, with the prediction that females will prefer conspecific calls. While on average female red deer preferred male red deer roars, two out of twenty females spent more time in close proximity to the speaker broadcasting male sika deer moans. We suggest that this absence of strict vocal preference for species-specific mating calls may contribute to the permeability of pre-zygotic reproductive barriers observed between these species. Our results also highlight the importance of examining inter-individual variation when studying the role of female preferences in species discrimination and intraspecific mate selection.     

Assessing the impact of past wapiti introductions into Scottish Highland red deer populations using a Y chromosome marker

As with other many game species in Europe, introductions of non-native stock to supplement Scottish red deer populations in an attempt to improve hunting trophy are well reported. These introductions included wapiti (Cervus elaphus canadensis or Cervus canadensis), a species two to three times heavier than the Scottish red deer. However, the effect of these past introductions of wapiti into Scottish red deer populations has not yet been assessed. In this study we sequenced a Y-chromosome marker (Zfy) from 104 collected in three neighbouring estates in the Scottish Highlands including one of the areas with the highest number of wapitis introduced in Scotland, and 45 red deer from an English deer park where introduction of wapiti and subsequent crosses with red deer were a common practice. Analyses revealed that all individuals presented red deer Y-chromosome sequences therefore suggesting a low impact of past introductions of wapiti in the populations under study. Sequencing and phylogenetic analyses of Zfy sequences for four additional deer species revealed that phylogenetic relationships were in agreement with previous mtDNA phylogenetic studies, and demonstrated the potential use of this marker to determine the direction of hybridization in F1 red-sika hybrids.     

Characterization of the erythrocyte superoxide dismutase allozymes in the deer Cervus elaphus

The cDNA sequences for Cu,Zn superoxide dismutase from two Cervus elaphus subspecies, North American wapiti and European red deer, were determined. The derived amino acid sequences showed two differences: residue 8 was Leu in wapiti and Met in red deer and residue 25 was His in wapiti and Asn in red deer. The extra positive charge at position 25 in the wapiti isoform accounted for its greater mobility towards the cathode during non-denaturing electrophoresis, a procedure widely used in the genetic analysis of deer. There was no difference in specific activity between the two Cu,Zn superoxide dismutase isoforms, but the wapiti isoform was slightly more susceptible to heat denaturation.     

A genetic study of sika (Cervus nippon) in the New Forest and in the Purbeck region, southern England: is there evidence of recent or past hybridization with red deer (Cervus elaphus)?

Sika Cervus nippon are native to Japan and East Asia but are now naturalized in many parks of the world, including Britain. In contrast to the substantial body of research on the extent of hybridization between sika and red deer Cervus elaphus in Scottish populations, there has been little genetic analysis of the English populations of sika. Sika in England still have a patchy and discontinuous distribution; populations are thus still genetically isolated and may be expected to show higher variability in genetic type. The current paper uses DNA-based techniques to explore the genetic composition of sika in the New Forest (Hampshire) and that of sika from the largest population in England, in the Purbeck region (Dorset). The study aims were to determine whether New Forest sika show signs of recent interbreeding with New Forest red deer populations, whether New Forest sika are genetically distinct from Purbeck sika, and whether New Forest and Purbeck sika show evidence of past hybridization or whether they are pure (non-hybrid) strains. Microsatellite analysis was used to compare the genetic profiles of individual deer. Results showed that sika and red deer in the New Forest were genetically distinct, indicating that there is no large-scale in situ hybridization occurring between these feral populations. In terms of overall genetic composition, there was no significant difference between the sika in the New Forest and Purbeck. However, a more detailed analysis found that New Forest sika showed a lower level of introgression with red deer compared with the Purbeck sika. We conclude that, overall, the New Forest sika deer do appear to be more genetically pure bred than the Purbeck sika.     

白唇鹿的染色体组型、C带和G带带型

白唇鹿产于我国西藏东部和四川西部。它和大熊猫一样,是我国特有的珍贵动物。有关它的染色体组型至今尚未见报道。我们于去年八月,对两头雄性白唇鹿外周血及茸缸淋巴细胞进行了培养,并作了染色体组型观察和C带及G带带型的观察。 一、材料和方法 北京动物园的两头准性白唇廘,一头10岁,一头4岁。 每一头先后采样二次。用装有司可林(Succinylcholinum chloratum)麻醉弹的麻醉枪击中鹿后,在颈静脉处,用含有1毫升肝素(浓度为50微     

Genetic analysis of erythrocyte superoxide dismutase polymorphism in the genus Cervus

A study of erythrocyte superoxide dismutase in Cervus dama, Cervus elaphus, Cervus nippon and Cervus elaphus x Cervus nippon hybrids has revealed a polymorphism of this enzyme system in Cervus elaphus as well as in the hybrid population. Genetic analysis suggests that this enzyme system is controlled by one gene locus with two codominant alleles. The allele frequencies allow a clear discrimination between the hybrid population and the red deer population, whereas the fallow deer are fixed for the allele which is most common in red deer. The comparison of the genotypic structures with the Hardy-Weinberg structure shows a slight excess of homozygotes in all populations, but the deviation is significant only for the hybrid population as well as one red deer population. Genetic variation within and differentiation between demes is quantified using different measures. Finally, some management implications of these first results are discussed.     

Mitochondrial DNA phylogeography of red deer (Cervus elaphus)

In order to understand the origin, phylogeny, and phylogeography of the species Cervus elaphus, we examined the DNA sequence variation of the mitochondrial cytochrome b gene of 51 populations of deer from the entire distribution area of Cervinae with an emphasis on Europe and Asia. Several methods, including maximum parsimony, maximum likelihood, and nested clade analysis, revealed that red deer originated from the area between Kyrgyzstan and Northern India. We found two distinct groups of red deer: a western group consisting of four subgroups and an eastern group consisting of three subgroups. Our mtDNA data do not support the traditional classification of red deer as only one species nor its division into numerous subspecies. The discrepancies between the geographical pattern of differentiation based on mtDNA cytochrome b and the existing specific and subspecific taxonomy based on morphology are discussed.     

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.     

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.     

Affinities between Cutifilaria (Nematoda: Filarioidea), parasites of deer, and Mansonella as seen in a new onchocercid, M. (C.) perforata n. sp., from Japan

A new dermal filarioid nematode, collected from Cervus nippon nippon (sika deer) on Kyushu Island, Japan, showed close affinities between the genera Cutifilaria and Mansonella (Onchocercidae: Onchocercinae): no buccal capsule, esophagus reduced to a thin fibrous tube, and female tail with four lappets. We propose Cutifilaria as a subgenus of Mansonella. Cutifilaria was distinguished from the five other subgenera, Mansonella, Tetrapetalonema, Esslingeria, Sandnema, and Tupainema, in having an area rugosa composed of transverse bands with tiny points, 14-16 papillae around the cloacal aperture, two prominent rhomboidal subterminal papillae, and a thick right spicule with spoon-shaped distal extremity. The host range of Mansonella was extended to ungulates by the addition of Cutifilaria, which appears to be derived from Tupainema, parasitic in Tupaioidea (insectivores), because of the similarity in their right spicules; Cutifilaria seems to have an Asiatic origin. M. (C.) perforata n. sp. was distinct from the sole other related species, M. (C.) wenki, a parasite of Cervus elaphus (red deer) in Europe, having a more complex right spicule with a sturdy terminal point and microfilariae with a bifid posterior end. In addition, almost all females had cuticular pores near the vulva, on the ventral line. The prevalence of microfiloriae and adults of M. (C.) perforata in the skin of sika deer was 38% and 21%, respectively.     

Introgression through rare hybridization: A genetic study of a hybrid zone between red and sika deer (genus Cervus) in Argyll, Scotland

In this article we describe the structure of a hybrid zone in Argyll, Scotland, between native red deer (Cervus elaphus) and introduced Japanese sika deer (Cervus nippon), on the basis of a genetic analysis using 11 microsatellite markers and mitochondrial DNA. In contrast to the findings of a previous study of the same population, we conclude that the deer fall into two distinct genetic classes, corresponding to either a sika-like or red-like phenotype. Introgression is rare at any one locus, but where the taxa overlap up to 40% of deer carry apparently introgressed alleles. While most putative hybrids are heterozygous at only one locus, there are rare multiple heterozygotes, reflecting significant linkage disequilibrium within both sika- and red-like populations. The rate of backcrossing into the sika population is estimated as H = 0.002 per generation and into red, H = 0.001 per generation. On the basis of historical evidence that red deer entered Kintyre only recently, a diffusion model evaluated by maximum likelihood shows that sika have increased at approximately 9.2% yr-1 from low frequency and disperse at a rate of approximately 3.7 km yr-1. Introgression into the red-like population is greater in the south, while introgression into sika varies little along the transect. For both sika- and red-like populations, the degree of introgression is 30-40% of that predicted from the rates of current hybridization inferred from linkage disequilibria; however, in neither case is this statistically significant evidence for selection against introgression.     

Effects of free-ranging,semi-captive and captive management on the acoustics of male rutting calls in Siberian wapiti <Emphasis Type="Italic">Cervus elaphus sibiricus</Emphasis>

In male European red deer Cervus elaphus, rutting calls that are responsible for male reproductive success are higher in fundamental frequency in captivity than in the wild. This study compares the acoustics of stag rutting calls among wild-living, semi-captive and captive stags within an Asian subspecies of C. elaphus, the Siberian wapiti Cervus elaphus sibiricus. Male Siberian wapiti rutting calls (bugles) were collected using automated recording systems in three populations (wild-living, semi-captive and captive), all originated from the Altai/Khakasian region of Central Siberia (Russia). Selected 435 bugles (145 per population) were analysed spectrographically for 14 variables of the bugle high (>1 kHz) fundamental frequency (g0) and scored for shape of g0 contour: trapeze, descending or saddle. Among bugles, 74.3% had the trapeze contour, 23.7% had the descending contour and 2.1% had the saddle contour. The additional low (<0.2 kHz) fundamental frequency (f0) was found in 76.1% of bugles, whereas deterministic chaos was found in 16.8% of bugles. Bugles of captive stags were shortest and highest in frequency. The captive management selectively affected only bugles with the trapeze contour, whereas bugles with descending contour remained unaffected by variations of deer holding regime. Stag rutting bugles are subspecies-specific and may therefore serve as acoustic indicator of subspecies for the Siberian wapiti among other Asian and American subspecies of wapiti.     

Parasites, diseases, and health status of sympatric populations of sika deer and white-tailed deer in Maryland and Virginia

In July 1981, investigations on parasites, diseases, and herd health status were conducted on sympatric populations of sika deer (Cervus nippon) and white-tailed deer (Odocoileus virginianus) from Blackwater National Wildlife Refuge (Maryland) and Chincoteague National Wildlife Refuge (Virginia) on the Delmarva Peninsula. Five adult deer of each species were collected from each location and subjected to thorough necropsy examinations and laboratory tests. White-tailed deer at both locations harbored protozoan, helminth, and arthropod parasites typically associated with this species throughout the southeastern United States. In contrast, sika deer at both locations harbored only light burdens of ticks, chiggers, and sarcocysts. Serologic tests for antibodies to seven infectious disease agents revealed evidence of exposure to bovine virus diarrhea (BVD) virus, infectious bovine rhinotracheitis virus, and parainfluenza3 virus in white-tailed deer, but only BVD virus in sika deer. At both locations the general health status of sika deer was superior to that of white-tailed deer.     

Prevalence of Neospora caninum and Toxoplasma gondii antibodies in wild ruminants from the countryside or captivity in the Czech Republic

In the Czech Republic, sera from 720 wild ruminants were examined for antibodies to Neospora caninum by screening competitive-inhibition enzyme-linked immunosorbent assay and confirmed by indirect fluorescence antibody test (IFAT); the same sera were also examined for antibodies to Toxoplasma gondii by IFAT. Neospora caninum antibodies were found in 14% (11 positive/79 tested) roe deer (Capreolus capreolus), 14% (2/14) sika deer (Cervus nippon), 6% (24/ 377) red deer (Cervus elaphus), 1% (2/143) fallow deer (Dama dama), 3% (3/105) mouflon (Ovis musimon), and none of 2 reindeer (Rangifer tarandus). Toxoplasma gondii antibodies were found in 50% (7/14) sika deer, 45% (169/377) red deer, 24% (19/79) roe deer, 17% (24/143) fallow deer, 9% (9/105) mouflon, and 1 of 2 reindeer. In 42 samples of wild ruminants that tested positive for N. caninum antibodies, 28 (67% of the positive N. caninum samples) reacted solely to N. caninum. This is the first evidence of N. caninum infection in mouflon, the first N. caninum seroprevalence study in farmed red deer, and the first survey of N. caninum in wild ruminants from the Czech Republic.