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Genetic studies of introduced non-native species are a valuable tool to investigate invasion history and pathways, source populations and multiple introductions of alien species, as well as evolutionary genetic changes following establishment in a new environment. We used a set of nine polymorphic microsatellites to analyse the population genetic structure of the introduced Asian shore crabHemigrapsus takanoialong European coasts. Our dataset covered the complete known European range of the species, including the most recent records from Great Britain and the southwestern Baltic Sea. The results showed a similarly high genetic diversity ofH. takanoithroughout Europe, and no indication of genetic bottlenecks during secondary spread, even in the most recently established populations. Analyses for population structure along geographic regions gave support for a separation between the Bay of Seine populations (northern France) and all other populations. Genetic differentiation within the North and Baltic Seas was more subtle and patchy, hinting to potential unrecognised introduction events, dispersal barriers and anthropogenic vector activity. The populations from the Baltic Sea and Great Britain clustered with the Wadden Sea populations, suggesting secondary introductions from the southeastern North Sea as likely invasion pathways. In summary, we suggest that a combination of anthropogenic secondary spread and the species’ reproductive biology have prevented a loss of genetic diversity during its ongoing expansion. We argue that genetic data depicting population status shortly after an introduction event—like the British and Baltic Sea populations ofH. takanoi—may provide important baseline data for investigations of genetic changes during establishment and adaptation processes.

We carried out a population genetic analysis of five southern African gemsbok (Oryx gazella) populations based on 530 bp of the mitochondrial control region and ten microsatellites in 75 individuals. Both markers show the high variability often observed in African bovids. Three of the populations which can be traced back to very small founding or current sizes do not show any signs of reduced variability compared to the remaining populations. The mitochondrial haplotypes form three distinct lineages which most likely originated in the Pleistocene when climate fluctuations led to periodical reduction and spreading of gemsbok habitat and which, today, are found throughout the distribution range. Bayesian microsatellite analyses yielded two groups, suggesting a more recent geographical differentiation following the admixture of the mtDNA lineages. Combining our sequences with available published data of the remaining oryx species allowed for a direct molecular comparison of O. gazella and O. beisa which have sometimes been considered a single species. The average genetic divergence between haplotypes from the two taxa was very high (39.9%), supporting their classification into two different species.

We analysed 131 common impala (Aepyceros melampus melampus) samples from two provinces in South Africa (Limpopo and KwaZulu-Natal) that are separated by the Drakensberg Mountain Range using sequences of the mitochondrial control region and seven polymorphic nuclear microsatellite loci. In line with earlier studies on bovid species, we found very high values of genetic diversity, particularly at the mtDNA locus with an overall nucleotide diversity of 3.6% and an overall haplotype diversity of 0.98. All statistical approaches confirmed a significant population differentiation between Limpopo and KwaZulu-Natal, suggesting that areas of unsuitable habitat caused by the presence of the Drakensberg Range and the Indian Ocean coastal belt act as a barrier to gene flow. Only few individuals with signs of admixed origin were indicative of translocations or rare migration between the two provinces. Combination of our mtDNA data set with those of previous studies on impala from south-western, southern and eastern Africa revealed the highest diversity in South Africa. This is in line with the hypothesis of a southern glacial refuge from which various African ungulate species spread northeast during the Holocene, although in the case of impala further analyses based on larger data sets will be necessary to definitively settle this question. Zusammenfassung Anhand genetischer Daten (mitochondriale Kontrollregion und sieben nukleare Mikrosatellitenloci) wurden 131 Impalas aus den durch die Drakensberge voneinander getrennten sudafrikanischen Provinzen Limpopo und KwaZulu-Natal untersucht. Im Einklang mit fruheren Studien an Boviden fanden wir eine sehr hohe genetische Variabilitat, insbesondere fur den mitochondrialen Locus mit Gesamtnukleotid- und Haplotypendiversitaten von 3.6% und 0.98. Samtliche statistischen Ansatze zeigten eine signifikante Differenzierung zwischen Populationen aus Limpopo und KwaZulu-Natal. Insbesondere die Drakensberge scheinen als Migrationsbarriere zu wirken. Wenige Individuen zeigten genomische Anteile beider Provinzen, was auf seltenen naturlichen Genflus oder Translokationen hinweist. Die Kombination unserer Daten mit denen fruherer Studien an Impalas aus dem sudwestlichen, sudlichen und ostlichen Afrika zeigten, das die Art in Sudafrika die hochste genetische Variabilitat aufweist. Dies steht in Einklang mit der Hypothese eines sudlichen Glazialrefugiums, aus dem sich verschiedene afrikanische Huftiere im Holozan nach Nordosten ausgebreitet haben.

As a consequence of persecution and habitat fragmentation, wildcats (Felis silvestris silvestris) in Western Europe have experienced a severe reduction in population numbers and sizes. The remaining wildcat populations are considered to be endangered by losses of genetic variability and by hybridisation with free-ranging domestic cats. To investigate genetic diversity within and among wild and domestic cat populations in Germany and to estimate the extent of gene flow between both forms, we analysed a total of 266 individuals. PCR-amplification and sequencing of 322 base pairs of a highly variable part of the mitochondrial control region (HV1) of 244 specimens resulted in 41 haplotypes with 31 polymorphic sites. Additionally, eight microsatellite loci were examined for those 244 cats. Moreover, a total of 46 wildcats and 22 domestic cats could be genotyped for 13 polymorphic out of 31 enzyme loci. Genetic variability in both groups was generally high. Variability in domestic cat populations was higher than in wildcat populations. Almost no differentiation between domestic cat populations could be found (FST for microsatellites=3%). In contrast, wildcat populations differed significantly from one another (FST for microsatellites=9.55%) Within the smaller wildcat populations, a reduction of genetic diversity was detectable with regard to the nuclear DNA. Wildcat and domestic cat mitochondrial haplotypes were separated, suggesting a very low level of maternal gene flow between both forms. In microsatellites and to a somewhat lesser extent in allozymes, wildcats and domestic cats showed distinct differentiation, suggesting an only low extent of past hybridisation in certain populations. The microsatellite data set indicated a significantly reduced effective population size (bottleneck) in the recent past for one German wildcat population.

We analysed 33 brown bears from the Romanian Carpathians and the Italian Apennines at sequences of the mitochondrial control region and nine polymorphic microsatellite loci with regard to genetic variability and haplotype distribution. The Italian brown bears were monomorphic for mtDNA sequences. The Romanian bears yielded the highest variability found so far in this species. Haplotypes of both previously identified mtDNA lineages (western and eastern) were found in Romania. In the eastern part of the Carpathians western and eastern haplotypes occurred sympatrically, the bears from the western part of the mountain range only exhibited western-type sequences. This pattern provides evidence of a mitochondrial phylogeographic break in the distribution of the eastern lineage within the Romanian Carpathians. Conservation implications of this finding are discussed.

Red deer (n = 149) from eight geographical locations, including the endangered endemic populations from the Tyrrhenian islands (Sardinia and Corsica), were analysed at eight polymorphic microsatellite loci. Two questions were addressed: (1) Is there a founder effect in the Corsican population, which was reintroduced to the island using Sardinian deer after the species’ extinction on Corsica? (2) What is the origin of the Tyrrhenian or Corsican red deer (Cervus elaphus corsicanus)? Our results showed signs of a founder effect for the red deer on Corsica in that these deer showed differentiation from the Sardinian population as measured by FST values, assignment tests (with and without a priori definition of populations) and individual-based dendrograms. Genetic variability, however, did not differ significantly between the two populations. With respect to the phylogeography of C. e. corsicanus we found that both deer from North-Africa and Mesola on the Italian mainland were genetically close to the Corsican red deer, but phylogenetic trees based on genetic distances were only poorly supported statistically. Among all populations studied the Mesola red deer showed the lowest distance values from Corsican red deer and yielded allele frequencies that were more similar to those of C. e. corsicanus than were those of North-African red deer. These results are in line with recent palaeontological and archaeozoological findings which suggest that the Corsican red deer is derived from small Italian red deer introduced from the mainland to Sardinia and Corsica during the Late Neolithic and just before the beginning of Classical Antiquity, respectively. They also suggest a possible recent introduction of Tyrrhenian red deer to North-Africa (rather than the other way around), thus accounting for the close genetic relationship (especially based on mitochondrial DNA) that has repeatedly been found between C. e. corsicanus and C. e. barbarus.

Barbary red deer Cervus elaphus barbarus from three areas in Tunisia (n=30) were analysed for genetic variability at 13 microsatellite loci and 680 bp of the mitochondrial control region. Barbary red deer, the only African deer taxon, are presently confined to a small area along the Tunisian–Algerian border and underwent a severe bottleneck in the middle of the 20th century with, on the Tunisian side, possibly only seven animals left. The genetic data showed signs of inbreeding and reflected the bottleneck event. Variability was on the lower end of the range previously reported for red deer (number of haplotypes=two, observed and expected heterozygosity 0.46 and 0.78, respectively) but not as low as might have been feared in view of the population's demographic history. The three sampling sites – among them the reserve at El Feidja from which, apart from Algerian immigrants, all extant Tunisian red deer originate – did not show any sign of genetic differentiation. The Tunisian red deer can thus probably be considered a single, genetically homogeneous population. Implications for management and conservation of the genetic findings are discussed.

A population genetic analysis based on sequences of the mitochondrial control region in 110 red foxes from five sampling localities in northern Serbia was carried out. The analysis yielded nine different haplotypes. Neither haplotype phylogeny nor their distribution was in accordance with the geographic location of the populations. In particular, the data failed to detect an unequivocal influence of the two big rivers, the Danube and the Tisza, separating the populations studied. Population differentiation was altogether low, without any relationship to the rivers as possible migration barriers. Although the possibility of foxes crossing the rivers over bridges or by swimming, thus keeping up gene flow, cannot be ruled out, it is most probable that the control region sequences are not sensitive enough to resolve small-scale population relationships but rather show patterns determined by stochastic processes such as genetic drift or lineage sorting.

In the present study, we analysed 18 red deer specimens from a small (N = 50) and isolated population in Schleswig-Holstein, northern Germany, with respect to variability at nine polymorphic microsatellite loci and 439 bp of the mitochondrial DNA control region. Several cases of brachygnathy (shortened lower jaw), commonly associated with inbreeding depression, have been recorded in the population. Genetic variability was very low compared with other European red deer populations including the neighbouring population from which the population under study was derived some 130 years ago. The effective population size was estimated to be seven individuals corresponding to an increase in inbreeding (or a loss of heterozygosity) of 7% each generation. This value is seven times higher than the theoretical threshold level up to which natural selection is believed to counteract the fixation of deleterious alleles in the gene pool. As a consequence, the population urgently needs genetic input from other populations to overcome the negative effects of random drift and inbreeding. To our knowledge, this study is one of the first to genetically analyse a red deer population showing strong signs of inbreeding depression.

Samples from 69 British red deer Cervus elaphus scoticus from seven populations in Scotland and England were analysed with respect to variability within and differentiation among stocks using 11 polymorphic microsatellite loci and 439 bp of the mitochondrial control region. The results clearly showed the effects of anthropogenic factors on British red deer. On the whole, variability values were within the species' reported range. The island population of Islay, Scotland, however, while showing average microsatellite variability, exhibited no mitochondrial variation at all. One microsatellite locus was monomorphic in three Scottish populations (Islay, Dunachton and Achnacarry). Overall and pairwise FST values indicate considerable differentiation among the populations studied, but Dunachton and Achnacarry, two adjacent populations free from recorded introductions, showed only a little differentiation and were paired in trees based on genetic distances. In terms of variability, no statistically significant differences were observed between island and mainland populations and the overall test of isolation by distance was negative. Possible reasons for the genetic patterns observed, such as differences in human impact on the populations, are discussed.

Summary The present study aimed at assessing genetic purity of black wildebeest ( Connochaetes gnou ) at Abe Bailey Nature Reserve, Gauteng Province, South Africa, using a multilocus microsatellite approach. Five loci were studied in black and blue ( C. taurinus ) wildebeest, the latter being a closely related species and known to produce hybrids with the morphologically very similar black wildebeest. In fact, the entire national black wildebeest population of South Africa potentially contains a significant proportion of introgressed blue wildebeest genes. In our case, eight out of 39 alleles were unique to black and 22 to blue wildebeest, with nine alleles shared between pure populations of the two species in line with their taxonomic proximity. A possible limited past introgression of blue wildebeest genes into the Abe Bailey population, corresponding to documents on population history, was only supported by the presence of a single allele otherwise exclusively found in samples of four pure blue but not in samples of two pure black wildebeest control populations. However, an assignment test and coefficients of population divergence did not support an extended introgression of C. taurinus alleles into the C .gnou population under study. Average heterozygosity at Abe Bailey proved to be intermediate between black and blue wildebeest, the latter species generally harbouring more genetic variation than the former owing to larger population sizes and the absence of population bottlenecks in historical times. The implications of our data are discussed with reference to the persistence of introgressed genes and the conservation of pure black wildebeest gene pools.

Allozyme, microsatellite and mtDNA (RFLP and sequence) data of European red deer populations were examined as to their capability of indicating anthropogenic influences such as the keeping of animals in enclosures, selective hunting for trophies translocation of specimens to improve trophy quality and habitat fragmentation. Deer in enclosures revealed considerable deviations of allele frequencies from isolation-by-distance expectations but no remarkable loss of genetic diversity. Particular allozyme genotypes were associated with antler morphology, and selective hunting was shown to alter allele frequencies in the expected direction. Habitat fragmentation is reflected by various kinds of genetic markers but due to the lack of information on population histories no unequivocal evidence on particular human activities could be obtained.

Despite an increasing acceptance in the biological community for sympatric speciation as a mode of species formation, well documented examples of sympatrically evolved ‘incipient species’ remain rare. The sympatric host races of apple maggot, Rhagoletis pomonella (Walsh), represent one of the most prominent case studies for sympatric speciation via a host shift. The European cherry fruit fly, R. cerasi (L.), shows strong ecological similarities to R . pomonella : (1) infestation of two different host plants, Lonicera xylosteum L. and Prunus avium L., and (2) divergent phenological and behavioral adaptations of flies on different hosts. The population genetic study presented here addresses whether the host associated populations of R. cerasi also represent genetically differentiated true host races. Out of a total of 29 allozyme loci examined, six were polymorphic and used to analyze six sympatric pairs of R. cerasi populations on Lonicera and Prunus from Switzerland and Germany. A direct comparison of allele frequencies between sympatric sites showed no pattern indicative of host races in R. cerasi . However, the hierarchical F-statistic for one locus, mannose 6-phosphate isomerase ( Mpi ), showed significant population differentiation that was in accordance with host race differentiation. Mpi is one of several loci that are also diagnostic for host race differentiation in R. pomonella . Results from Mpi suggest the formation of sympatric host races in R. cerasi , but additional polymorphic markers are necessary.

Five south-European red deer populations from Spain, mainland Italy (Val di Susa and Tarvis), Sardinia and Bulgaria were analysed with regard to microsatellite loci and the mtDNA control region to cast light on the phylogeographic origin of the Corsican red deer which is restricted to Corsica and Sardinia. Whilst according to mtDNA haplotypes Sardinian red deer showed the closest affinity to the Spanish population, microsatellite analyses yielded completely different results in that Sardinia and Spain exhibited the greatest genetic distance whereas Tarvis was the population most closely related to Sardinia. These results are discussed including evidence as to the oldest fossil red deer remains on Sardinia. The latter could play a pivotal role regarding the decision between introduction of red deer on Corsica and Sardinia through humans in Holocene or natural settlement in Upper Pleistocene times.

Tiedemann R., Kurt F., Haberhauer A. and Hartl G. B. 1998. Differences in spatial genetic population structure between African and Asian elephant (Loxodonta africana, Elephas maximus) as revealed by sequence analysis of the mitochondrial Cyt b gene. [In: Ecological genetics in mammals III. G. B. Hartl and J. Markowski, eds). Acta Theriologica, Suppl. 5: 123-134. To assess spatial genetic population structure of both extant elephant species, we investigated sequence variation in 369bp of the mitochondrial Cyt b gene in 23 specimens of African elephant Loxodonta africana (Blumenbach, 1797) from three regions in the Southern part of Africa. In an integrated analysis, these results were compared to data of a previous study, where the same gene region had been analysed in 53 Asian elephants Elephas maximus (Linnaeus, 1758) from 5 different regions on Sri Lanka and Asian mainland. In Loxodonta, 14 polymorphic sites defined 6 different mitochondrial haplotypes with a mean sequence divergence of 2.085%. In Elephas, 6 polymorphic sites defined 8 different haplotypes with a mean sequence divergence of 0.942%. Compared to other mammals, genetic variation is high in Loxodonta and moderate in Elephas. The difference in genetic variation among the species could be explained either by a Pliocene bottleneck in Elephas or by different long term effective population sizes. In Elephas, a star like phylogeny of haplotypes was found, indicative of a population expansion after a bottleneck. In Loxodonta, very divergent mtDNA lineages coexisted, suggesting the absence of any bottleneck in population history. Within regional subpopulations, both species showed similar mean haplotype diversities, while mean nucleotide diversity within regions was higher in Loxodonta than in Elephas. This suggest larger long-term effective population sizes in Loxodonta, while short-term effective population sizes are presumably similar in both species. Spatial genetic population structure in Loxodonta is mainly determined by isolation-by-distance, while in Elephas it is impacted by human translocation. Human translocation might have prevented isolated small Elephas populations from severe genetic depletion. Institut fur Haustierkunde, Christian-Albrechts-Universitat zu Kiel, Olshausenstrase 40, D-24118 Kiel, Germany

The Quaternary dispersal of the red fox Vulpes vulpes in the Mediterranean area was evaluated through the study of allelic variation at 45 enzyme loci in 120 individuals from 10 sampling sites. A 375 bp fragment of the mitochondrial cytochrome b gene was also sequenced in a total of 41 specimens from the same sampling locations. Nine allozyme loci were polymorphic. The proportion of polymorphic loci per population (P) ranged from 0 to 15.6%, and expected average heterozygosity (H) from 0 to 4.4%. A total of 18 different Cyt b haplotypes were detected. Most of them were confined to only one population. Both allozyme and mtDNA data implied that our fox populations were genetically fairly isolated from one another, suggesting low gene flow between them. This isolation should be of comparatively recent origin according to the slight differentiation among Cyt b haplotypes. Fox populations appeared to belong to two genetically distinct groups. With a mean value of Nei’s D = 0.024, genetic distance between these groups was similar to that detected at subspecies level in taxa of large mammals. This pattern may have originated from different colonization waves during Quaternary glaciations and deglaciations. Red foxes from Sardinia were more closely related to the Bulgarian foxes than to the Iberian ones. However, repeated introductions to Sardinia probably also occurred from Central Italy and Spain, as suggested by the presence of haplotype A and a typical Central Italian allele, Ck-290

The red fox (Vulpes vulpes) is a highly adaptable omnivorous mammal distributed across all continents on the northern hemisphere. Although the red fox is present throughout Europe, where it plays an important ecological and socio-economic role not only as a game species but also as a rabies reservoir, few studies have examined its population-level mitochondrial DNA variability. In this study, 27 mitochondrial DNA control region haplotypes were identified in 229 red fox samples taken from four regions in Croatia. Haplotype diversity of Croatian red foxes (0.901) was found to be among the highest of all European red fox populations studied to date. Genetic differentiation among regions was quite low, and statistically significant estimates of differentiation were obtained only when comparing the population from the peninsular region of Istria with the three continental populations. It seems that landscape barriers like rivers and small mountains do not restrict gene flow among foxes in the continental part of Croatia, while the combination of a narrow land bridge and altitudes exceeding 1, 000 m limit fox migration between Istria and the rest of the continent. Better understanding of small-scale population structure will require analysis of highly variable nuclear markers like microsatellites.

In order to examine a previously hypothesized influence of selective hunting on allele frequency changes at some regularly polymorphic allozyme loci in red deer (Cervus eluphus), antler characters, serving as criteria for culling, were examined in relation to electrophoretic variation in two free-ranging populations of the Vosges, Eastern France, and an enclosure in Central France. When homozygous for the allele Idh-2125, stags ≥ 2 years old had a significantly higher number of antler points (NAP). When homozygous for the allele Acp-2100, stags older than 5 years had antlers that were significantly larger for a number of traits (NAP, main beam length and circumference, coronet circumference, brow tine length). Among younger stags, all antler traits in Acp-2100 homozygotes were significantly smaller than in carriers of the alternative allele, Acp-285. Our data suggest the presence of at least two independent genetic components (one associated with early development of a high NAP, the other with generally large antler size in adults), affecting antler expression in red deer. Those genetic components, possibly major genes which are chromosomally linked with the allozyme loci studied, compensate or reinforce each other in their phenotypic effects. By playing a role in balancing benefits and costs of male reproductive success, they may be part of a genetic mechanism enabling the rapid adaptation of a population to various environmental and demographic conditions. The three populations studied originate from one another, and, based on an assessment of effective population sizes, it could be demonstrated that selective hunting for antler shape has changed allelic frequencies at the associated marker loci within a few generations.

To investigate genetic diversity in captive Asian elephants (Elephas maximus), a total of 26 individuals from zoos in the German cities of Berlin, Hamburg, and Leipzig, and from Zurich, Switzerland, were investigated for electrophoretic variation at 44 presumptive blood protein and enzyme coding loci. The proportion of polymorphic loci (P = 4.6%) and average heterozygosity (H = 1.4%) in the specimens studied were very low. The distribution of alleles and genotypes suggested some genetic differentiation among the source populations of our elephants, most of which were wild born. Since our results are in accordance with those of a previous survey on wild populations, low genetic variation in the Asian elephant is not only associated with captive propagation. Together with differences in the incidence of alleles among populations, it is probably largely due to low effective sizes of wild Asian elephant stocks and to species-specific breeding tactics. Standard karyotype analysis was carried out for 11 individuals, and in some cases, heterochromatin staining by Giemsa C-banding and reverse fluorochrome staining were performed. Heterochromatin polymorphism was found in four chromosome pairs, including two satellited pairs, and is considered an important marker for population genetic investigations. One of the 11 individuals studied was found to be heterozygous for a centric fusion of two small acrocentrics. The implications of our results for breeding programs of Asian elephants are discussed. © 1995 Wiley-Liss, Inc.

Genetic variation in an isolated brown bear population of the Western Carpathians was studied by electrophoretic analysis of 51 presumptive allozyme loci. In spite of a severe population bottleneck at the beginning of the 1930s (40 survivors), average heterozygosity (H=5.3%) was within the range commonly found in mammals and the mean number of alleles per polymorphic locus (Ap=3, over five polymorphic loci) was very high. Possible reasons for the maintenance of high allelic variation are discussed.

A total of 469 brown hares Lepus europaeus Pallas, 1778 from 20 sampling sites in Austria were examined for genetic diversity within and among populations by means of horizontal starch gel electrophoresis. Fourteen out of 54 presumptive structural loci were polymorphic, one of which was excluded from further population genetic analyses due to the occurrence of a null-allele.

Biochemical-genetic variation was studied in springbok Antidorcas m. marsupialis (Zimmermann, 1780) from a large (N>2000) "wild" population (n=24) and a small (30>N>20) isolated farm population (n=10) using electrophoretic allozyme analysis. Springbok showed polymorphisms at eight out of 46 loci.

We analyzed 121 golden jackals (Canis aureus) from six sample sites in Serbia with regard to genetic variability and differentiation as revealed by mitochondrial control region sequences and eight nuclear microsatellite loci. There was no variation at all in the mtDNA sequences, and nuclear variability was very low (average observed and expected heterozygosity of 0.29 and 0.34, respectively). This is in line with the considerable recent range expansion of this species in the Balkans and indicates a strong founder effect in the recently established Serbian population. We did not find evidence of differentiation between the northeastern jackals and those from the plain of Srem or those in between. F-statistics and Bayesian Structure analyses, however, were indicative of a low degree of overall differentiation in the Serbian population. A vagrant Austrian jackal that was also analyzed was genetically indistinguishable from its Serbian conspecifics.

The possibly distinct Carpathian red deer was compared genetically to other European populations. We screened 120 red deer specimens from Serbia, the Romanian lowland and the Romanian Carpathians for genetic variability using 582 bp of the mitochondrial control region and nine polymorphic nuclear microsatellite loci. The study aimed at a population genetic characterization of the Carpathian red deer, which are often treated as a distinct subspecies (Cervus elaphus montanus). The genetic integrity of the Carpathian populations was confirmed through the haplotype distribution, private alleles and genetic distances. The Carpathian red deer are thus identified as one of the few remaining natural populations of this species, deserving special attention among game and conservation biologists. The history of the populations studied, in particular the introduction of Carpathian red deer into Romanian lowland areas in the 20th century, was reflected by the genetic data.

European beavers, Castor fiber, were recently reintroduced to Austria and Bavaria and the population has been increasing rapidly. In 1996 we began livetrapping animals for reintroduction programs in neighboring countries. There are currently North American beavers, C. canadensis, in Bavaria, and a small number were inadvertently released in Austria 20 years ago. It is critical to be able to discriminate between North American beavers and European beavers that are trapped as part of any reintroduction program in Europe. For our reintroduction work we need to definitatively determine the species of each trapped beaver so no new North American beavers are released. It is also critical to be able to discrimiate between the two species so the distribution of North American beavers in our region may be determined. Discrimination between the two species by morphometric methods is difficult and not definitive using live animals. Preparing chromosomes for karyotypic studies takes time and only works properly with fresh blood. Therefore, we developed a method that uses tissues from either live or dead compare two North American beavers and two European beavers, we only observed variation at the Esterase-D (E.C.3.1.1.1) locus out of the 14 loci examined. To further test our new method of discriminating between the two species we used either animals whose species was already known, or confirmed our results by preparing additional karyotypes. We screened blood and kidney samples from 37 individuals of C. fiber and from 9 individuals of C. canadensis for variation of the Es-D locus by horizontal starch gel electrophoresis. All C. fiber exhibited a fast migrating allele and all C. canadensis a slow migrating allele at this locus. Hence, the Es-D locus can be interpreted as a differential-diagnostic for the two species.

Thirty-three random decanucleotide primers were tested for polymorphisms in a fenced Italian population of fallow deer Dama dama (Linnaeus, 1758) and 23 primers provided reproducible amplification patterns.

haplotypes ranged from 0.05% to 0.25%. One chamois from the High Tatra (subspecies R. r. tatrica) was examined and showed the standard haplotype found in R. r. rupicapra. MtDNA in chamois from Catalunya, belonging to R. pyrenaica pyrenaica, was polymorphic for two haplotypes not found in any population of R. rupicapra. Mean nucleotide divergence among haplotypes found in R. rupicapra and R. pyrenaica was 0.56% (SD = 0.16%). Based on this value, an estimated divergence time of about 280 000 years suggests that the mtDNA lineages of R. rupicapra and R. pyrenaica separated prior to the Riss glacial in the later Pleistocene.