Phylogeography of red deer (Cervus elaphus) in Europe

Aim To investigate the phylogeographical patterns of red deer (Cervus elaphus) in Europe, and to disentangle the influence of ancient (e.g. Pleistocene ice ages) from more recent processes (e.g. human translocations). Location Europe. Methods In this study we provide by far the most extensive analysis of genetic structure in European red deer, based on analyses of variation at two mitochondrial markers (cyt b and D‐loop) in a large number of individuals from 39 locations. Relationships of mitochondrial DNA haplotypes were determined using minimum spanning networks and phylogenetic analyses. Population structure was examined by analyses of molecular variance. Historical processes shaping the present patterns were inferred from nested clade analysis and nucleotide diversity statistics. Results Within Europe, we detected three deeply divergent mitochondrial DNA lineages. The three lineages displayed a phylogeographical pattern dividing individuals into western European, eastern European and Mediterranean (Sardinia, Spain and Africa) groups, suggesting contraction into three separate refugia during the last glaciation. Few haplotypes were shared among these three groups, a finding also confirmed by F_ST values. Calculations of divergence times suggest that the groups probably split during the Pleistocene. Main conclusions The observed pattern is interpreted to result from isolation in different refugia during the last glaciation. The western and eastern European lineages could be linked to an Iberian and Balkan refugium, respectively. The third lineage might originate from a Sardinian or African refugium. We link local phylogeographical patterns observed in Europe to the post‐glacial recolonization process, shaped by the geographical localization of refugia and barriers to gene flow. Regardless of the importance of red deer as a game species and the tradition of translocating red deer in Europe, we detected few individuals that did not match the trichotomous pattern, suggesting that translocations have occurred mainly at smaller spatial scales.     

Variations in bark-stripping by red deer Cervus elaphus across Europe

• 1 The literature on bark‐stripping by red deer Cervus elaphus in Europe is reviewed to reveal quantitative variation in this behaviour and relate it to deer density and local characteristics such as dominant tree species, occurrence of artificial feeding, altitude, region and size of the study site. We also review the importance of bark in red deer diets over the seasons and discuss the causes of bark‐stripping, focusing on the significance of bark as food.
• 2 Over the 36 sites examined, the rate of bark‐stripping was highly variable (from 0 to 84% of susceptible trees debarked), with less damage in Scotland than in other European sites for which bark‐stripping rates were higher at high red deer density. Altitude, the size of the study site, the number of dominant tree species and the occurrence of artificial feeding do not significantly relate to the rate of bark‐stripping.
• 3 Bark sometimes made up a large proportion of red deer diet (> 10%), especially in areas with severe winters (high levels of snow), whereas in study sites with mild winters, bark was practically not eaten at all.
• 4 These results suggest that severe bark‐stripping could be related to a reduction in food resource availability. This food availability hypothesis needs to be better documented, dealing particularly with the possible interaction between food availability and red deer density.

     

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.     

Molecular identification of seven Sarcocystis species in red deer (Cervus elaphus) from Lithuania

The diaphragm muscles of 77 free-ranging red deer (Cervus elaphus) were examined for Sarcocystis species in Lithuania. Sarcocysts were detected in 61 out of 77 (79.2%) animals investigated. A total of 60 isolated sarcocysts were identified to species using subunit I of cytochrome c oxidase (cox1) sequence analysis. Overall, seven species, S. entzerothi, S. hjorti, S. iberica, S. linearis, S. pilosa, S. truncata and S. venatoria, were confirmed in Lithuanian red deer. Sarcocystis entzerothi was reported in red deer for the first time. Previously this species was shown to use sika deer as well as roe deer and fallow deer as an intermediate host. Based on cox1, with the addition of the current data, altogether 13 Sarcocystis species have so far been shown to use red deer as an intermediate host. Species detected in red deer demonstrated considerable differences in intraspecific genetic variation at cox1. Genetic distances between different samples of S. hjorti and S. linearis were calculated using principal coordinates analysis (PCoA), implying molecular divergence of same Sarcocystis species using different hosts in the same geographical area and divergence of those employing same intermediate host species from different areas.     

Onchocercosis in red deer (Cervus elaphus) from Spain.

Onchocercosis, caused by Onchocerca flexuosa, was observed in red deer (Cervus elaphus) from Spain for the first time. Adult specimens of P. flexuosa were found in nodules in subcutaneous tissues in 42 of 125 (33%) red deer between October 1994 and September 1995; intensity of infection +/- SD was 3.93 +/- 5.26 nodules per infected host. A clear seasonal pattern in the distribution of nodules was observed, with higher values of prevalence and intensity in fall and winter in contrast to spring and summer. Significant differences were found among age groups in prevalence, but not in the mean intensity. No differences in infection were apparent between male and female adult red deer.     

Freemartinism in red deer (Cervus elaphus L.)

Several mixed-sex twins were produced by red deer treated with progesterone and pregnant mare's serum gonadotrophin. Of seven females karyotyped, two were 68,XX/68,XY chimeras. One male had the same chimeric karyotype as its female co-twin. These are the first reported cases of freemartinism in deer.     

Malignant melanoma in a captive red deer (Cervus elaphus elaphus)

We report a case of malignant melanoma in a captive red deer (Cervus elaphus elaphus). The primary lesion was on the plantar aspect of the right pastern and infiltrated the surrounding skin, but not underlying tendons or bone. Diffuse metastasis to the regional lymph nodes had occurred. Histologically, lesions were characterized by large numbers of variably pigmented epithelioid cells with a high degree of nuclear atypia and many mitotic figures. This is the first reported case of this neoplasm in red deer.     

Physiological effects of hunting red deer (Cervus elaphus).

When red deer (Cervus elaphus) were hunted by humans with hounds the average distance travelled was at least 19 km. This study of 64 hunted red deer provides the first empirical evidence on their state at the time of death. Blood and muscle samples obtained from hunted deer after death were compared with samples from 50 non-hunted red deer that had been cleanly shot with rifles. The effects on deer of long hunts were (i) depletion of carbohydrate resources for powering muscles, (ii) disruption of muscle tissue, and (iii) elevated secretion of beta-endorphin. High concentrations of cortisol, typically associated with extreme physiological and psychological stress, were found. Damage to red blood cells occurred early in the hunts; possible mechanisms are discussed. Taken together, the evidence suggests that red deer are not well-adapted by their evolutionary or individual history to cope with the level of activity imposed on them when hunted with hounds.     

Genetic structure of the Danish red deer (Cervus elaphus)

The red deer (Cervus elaphus) population in Denmark became almost extinct in recent historical times due to over‐hunting. The species has subsequently recovered within remote areas, but non‐Danish individuals have been introduced at several localities. To assess genetic structure, past demographic history, and the possibility of a still existing original stock, we analysed 349 specimens from 11 geographically separate areas and from three enclosed areas, genotyping 11 microsatellite loci. Moreover, an 826‐bp fragment of the control region of the mitochondrial DNA was sequenced for 116 recent specimens and seven museum specimens. There was a significant difference in mean expected heterozygosity (H_E) between the three enclosed areas and the 11 unenclosed areas. Significant departures from Hardy–Weinberg equilibrium were observed in the three enclosed areas and in nine of the unenclosed areas. The overall degree of genetic differentiation among all 14 areas was significant (F_ST = 0.09, P < 0.01), primarily because the mean pairwise F_ST for the three enclosed areas was significantly higher than that for the 11 unenclosed areas. A Bayesian clustering procedure detected three genetically distinct populations and indicated reduced gene flow between the enclosed and unenclosed areas. The individuals in the unenclosed areas show genotypic mixture, presumably as a result of gene flow among them. Markov Chain Monte Carlo simulations, based on the genealogical history of the microsatellite alleles, suggest a drastic decline in the effective population size of the enclosed areas some 188–474 years ago. Mitochondrial DNA analysis of the recent specimens showed seven haplotypes. Individuals from the enclosed Jægersborg Dyrehave contain haplotypes that occur all over Denmark and also are found in Western Europe. A close relationship between Scandinavian and Western European red deer is most likely. Only individuals from the unenclosed Lindenborg Estate and the enclosed Tofte Skov did not group with any other Danish individuals. As six of seven museum specimens had haplotypes also found in modern Danish samples, the current population of red deer in Denmark is genetically close to the original Danish red deer. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 688–701.     

Abomasal nematodes of the red deer Cervus elaphus in north-eastern Italy

The composition of the abomasal helminth fauna and parasite diversity were studied in 298 red deer collected during 1997-2000 from three different sectors (Bolzano, Trento and Sondrio provinces) of the Stelvio National Park, one of the main protected areas of north-eastern Italy. The association between parasite burdens and geographical areas of the hosts was assessed using the negative binomial regression. A variety of abomasal helminths, both host specific and generalist, was found in all sectors. The most commonly observed parasites were the Spiculopteragia spiculoptera morph spiculoptera and Ostertagia leptospicularis morph leptospicularis, with prevalences of 79.5% and 40.9%, respectively. The minor morphs S. spiculoptera morph mathevossiani (prevalence 31.9%) and O. leptospicularis morph kolchida (18.8%) occurred at lower prevalences. Teladorsagia circumcincta morph circumcincta, Marshallagia marshalli morph marshalli and Haemonchus contortus were rarer, at prevalences of 1, 1.3 and 1.3%, respectively. Deviance analysis of the negative binomial regression model shows that the geographical area is significantly related to parasite burdens (P = 0.001). Prevalences of hosts with parasites were greater in the Sondrio (odds ratio = 1.31; 95% confidence interval: 0.16-10.85) and smaller in the Trento (odds ratio = 0.62; 95% confidence interval: 0.20-1.96) sectors with respect to Bolzano, but these differences were not statistically significant. Possible cross-infections by more generalist parasites between wild and domestic animals were also suggested, as deer and domestic ruminants (Bovinae, Caprinae and Ovinae) used the same feeding areas of the park sectors.     

Hypodermosis in the red deer Cervus elaphus in Cordoba, Spain

The prevalence of red deer hypodermosis and the life cycle of Hypodermia diana Brauer 1858 in three provinces of the south of Spain, Córdoba, Jaen and Ciudad Real, were studied by inspecting 254, thirty-eight and thirty-five deer from each province respectively. The prevalence of infestations was: Córdoba, 87.75%; Jaen, 92.10%, Ciudad Real, 91.42%. From this we deduced an overall prevalence of 88.67%, comprising 88.23% in male deer and 89.96% in female deer. The intensity of the parasitism in 213 of the 245 animals from Córdoba varied between one warble per animal to more than 100 in 28 of the inspected animals. The chronology of the life cycle during the hunting period (October to the end of February) was investigated.     

Variation of some serum proteins in red deer, Cervus elaphus L

Various electrophoretic techniques, immunoblotting and inhibitions of trypsin and chymotrypsin were used to study the variability of serum proteins in farmed red deer, Cervus elaphus L., of Czechoslovakian origin. Easily interpretable polymorphisms were observed in transferrin (variants A, B1, B2, C) and vitamin D binding protein, GC (variants D, F, I, S). Great variability was observed in the protease inhibitors PI2, PI3, PI4, PI5, and PI8 and in unidentified zones in the vicinity of albumin, but no genetical or physiological interpretation for this variability is yet available. Haemopexin, alpha 1 glycoprotein, protease inhibitors PI1, PI6 and PI7 were monomorphic.     

Morphological observation of antler regeneration in red deer (Cervus elaphus)

Deer antler offers a unique opportunity to explore how nature solves the problem of mammalian appendage regeneration. Annual antler renewal is an example of epimorphic regeneration, which is known to take place through initial blastema formation. Detailed examination of the early process of antler regeneration, however, has thus far been lacking. Therefore, we conducted morphological observations on antler regeneration from naturally cast and artificially created pedicle/antler stumps. On the naturally cast pedicle stumps, early antler regeneration underwent four distinguishable stages (with the Chinese equivalent names): casting of previous hard antlers (oil lamp bowl), early wound healing (tiger eye), late wound healing and early regeneration (millstone), and formation of main beam and brown tine (small saddle). Overall, no cone-shaped regenerate, a common feature to blastema-based regeneration, was observed. Taken together with the examination on the sagittal plane of each regenerating stage sample, we found that there are considerable overlaps between late-stage wound healing and the establishment of posterior and anterior growth centers. Observation of antler regeneration from the artificially created stumps showed that the regeneration potential of antler remnants was significantly reduced compared with that of pedicle tissue. Interestingly, the distal portion of a pedicle stump had greater regeneration potential than the proximal region, although this differential potential may not be constitutive, but rather caused by whether or not pedicle antlerogenic tissue becomes closely associated with the enveloping skin at the cut plane. Antler formation could take place from the distal peripheral tissues of an antler/pedicle stump, without the obvious participation of the entire central bony portion. Overall, our morphological results do not support the notion that antler regeneration takes place through the initial formation of a blastema; rather, it may be a stem cell-based process.     

First report of Elaphostrongylus cervi in Spanish red deer Cervus elaphus hispanicus

Elaphostrongylus cervi Cameron, 1931 was identified in six Cervus elaphus hispanicus sampled in Cuenca, central Spain. A total of 23 adult worms were found in the central nervous system with a mean of 3.8. Although E. cervi is reported to be widespread in cervids, this is the first time it has been recorded in Spanish red deer.