Christian Pitra

When the American sea sturgeon swam east

The two species of Atlantic sea sturgeon on either shore of the North Atlantic, Acipenser sturio in Europe and A. oxyrinchus in North America, probably diverged with the closure of the Tethys Sea and the onset of the North Atlantic Gyre 15–20 million years ago, and contact between them was then presumably precluded by geographic distance. Here we present genetic, morphological and archaeological evidence indicating that the North American sturgeon colonized the Baltic during the Middle Ages and replaced the native sturgeon there, before recently becoming extinct itself in Europe as a result of human activities. In addition to representing a unique transatlantic colonization event by a fish that swims upriver to spawn, our findings have important implications for projects aimed at restocking Baltic waters with the European sturgeon.

Fish populations: When the American sea sturgeon swam east

The two species of Atlantic sea sturgeon on either shore of the North Atlantic, Acipenser sturio in Europe and A. oxyrinchus in North America, probably diverged with the closure of the Tethys Sea and the onset of the North Atlantic Gyre 15–20 million years ago, and contact between them was then presumably precluded by geographic distance. Here we present genetic, morphological and archaeological evidence indicating that the North American sturgeon colonized the Baltic during the Middle Ages and replaced the native sturgeon there, before recently becoming extinct itself in Europe as a result of human activities. In addition to representing a unique transatlantic colonization event by a fish that swims upriver to spawn, our findings have important implications for projects aimed at restocking Baltic waters with the European sturgeon.

An exceptional case of historical outbreeding in African sable antelope populations

Empirical investigations of intraspecific outbreeding and subsequent introgressive hybridization in natural populations are rare, particularly among conspecific populations of large mammals. Using mitochondrial DNA data [partial control region (496 basepairs — bp) and cytochrome b gene (343 bp) sequences analysed from 95 individuals representing 17 sampling locations scattered through the African miombo (Brachystegia) woodland ecosystem] and phylogeographical statistical procedures (gene genealogy, nested cladistic and admixture proportion analyses), we (i) give a detailed dissection of the geographical genetic structure of Hippotragus niger; (ii) infer the processes and events potentially involved in the population history; and (iii) trace extensive introgressive hybridization in the species. The present‐day sable antelope population shows a tripartite pattern of genetic subdivision representing West Tanzanian, Kenya/East Tanzanian and Southern Africa locations. Nested clade analysis revealed that past allopatric fragmentation, caused probably by habitat discontinuities associated with the East African Rift Valley system, together with intermediary episodic long‐distance colonization and restricted, recurrent gene flow have played an predominant role in shaping the extent of maternal genetic diversity (10.4%) and population structure. An extensive (average rate of admixture = 20.0%), but geographically circumscribed and unidirectional hybridization event in the past was inferred, resulting in an extreme (the highest discovered so far in mammals) intraspecific difference of 18.2% among morphologically monotypic sable antelopes from West Tanzania. The results are used to provide an evolutionary framework within which taxonomic implications and conservation decisions can be evaluated.

Species identification of Oetzi's clothing with matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry based on peptide pattern similarities of hair digests

Identification of ancient biological samples from the 1991-discovered and more than 5300-year-old Tyrolean mummy, also called iceman or Oetzi, is very difficult. The species of origins of four animal-hair-bearing samples of the accoutrement of the mummy not yet diagnosed were identified by a special proteomics method. Ha 43/91/130 and Ha 6/91, two samples from his coat, and Ha 5/91, a sample from his leggings, were assigned to sheep. The upper leather of his moccasins, Ha 2/91, was made from cattle. Despite the enormous age of these samples with partial (bio)chemical alterations, reliable identification was possible using a recently developed matrix-assisted laser desorption/ionization time-of-flight mass spectrometric ((MALDI-TOF MS)-based analytical method. The method is exclusively based on the analysis of proteins and uses minute amounts of peptides directly derived from tryptic hair digests without any separation or enrichment steps. Unknown species are identified by comparison of their peptide ion patterns with known spectra stored in existing databases. Hereby, the correlation distance, a form of Euclidean distance, and deduced parameters are used to measure similarities. If more than one potential hit remains, specific diagnostic peptide ions are used to stepwise exclude incorrect matches. These ions are specific for orders, families, subfamilies/genera and/or even species. Peptide mass fingerprinting data combined with those from collision-induced dissociation spectra (combined MS & MS/MS) were used for interpretation with the MASCOT search engine and the NCBI database to find the potential parentage of hair proteins. For this technique, selected precursor ions were identified as specific diagnostic peptide ions.

Population subdivision in Europe’s great bustard inferred from mitochondrial and nuclear DNA sequence variation

A continent‐wide survey of sequence variation in mitochondrial (mt) and nuclear (n) DNA of the endangered great bustard (Otis tarda) was conducted to assess the extent of phylogeographic structure in a morphologically monotypic bird. DNA sequence variation in a combined 809 bp segment of the mtDNA genome from 66 individuals from the last six breeding regions showed relatively low levels of intraspecific sequence diversity (π = 0.32%) but significant differences in the regional distribution of 11 haplotypes (ΦST = 0.49). Despite their exceptional potential for dispersal, a complete and long‐term historical separation between the populations from the Iberian Peninsula (Spain) and mainland Europe (Hungary, Slovakia, Germany, and Russia) was demonstrated. Divergence between populations based on a 3‐bp insertion–deletion polymorphism within the intron region of the nuclear CHD‐Z gene was geographically concordant with the primary subdivision identified within the mtDNA sequences. Inferred aspects of phylogeography were used to formulate conservation recommendations for this endangered species.

GENETIC STRUCTURE OF POPULATIONS OF EUROPEAN BROWN HARE: IMPLICATIONS FOR MANAGEMENT

Abstract During the last 2 decades, European brown hare (Lepus europaeus) populations have declined considerably. We evaluated whether this decline could have been associated with diminished overall fitness due to reduced genetic variability. We also estimated the extent of population differentiation. We typed 307 hares from 21 localities for 5 microsatellite loci and the mitochondrial d-loop. Analysis of molecular variance revealed a high degree of genetic variability, matrilinearily structured populations, male-biased gene flow, and lack of inbreeding. We did not identify any geographical or anthropogenic barriers to gene flow. Because extant populations are small and susceptible to random genetic drift, we recommend changes to current population management practices and periodic genetic surveys.

Great bustard population structure in central Spain: concordant results from genetic analysis and dispersal study

We found significant sex differences in the mtDNA genetic structure and dispersal patterns of great bustards in a population of 11 breeding groups, ‘leks’, in central Spain. The analysis of genetic distances showed that the female population was divided into three groups of leks separated by ca. 50 km, whereas male haplotypes were randomly distributed among leks. Genetic distances among pairs of leks were positively correlated with geographical distances in females but not in males. While female haplotype distributions were homogeneous among leks at close distances, differences in male genetic structure were highly variable even between two close leks. These results from genetic analyses were concordant with those from a radiotracking study on natal dispersal. Natal dispersal distances were higher in males than in females. Also, the frequency of movement of a female between two leks was positively correlated with their genetic affinity and geographical proximity. In males, the frequency of movement was correlated with geographical proximity but not with genetic affinity. Males dispersed among genetically unrelated leks, contributing to keep nuclear genetic diversity in the population, whereas females tended to be philopatric. These results suggest that isolation–by–distance influences the distribution of maternal lineages at a regional level.

Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry combined with multidimensional scaling, binary hierarchical cluster tree and selected diagnostic masses improves species identification of Neolithic keratin sequences from furs of the Tyrolean Iceman Oetzi

The identification of fur origins from the 5300-year-old Tyrolean Icemans accoutrement is not yet complete, although definite identification is essential for the socio-cultural context of his epoch. Neither have all potential samples been identified so far, nor there has a consensus been reached on the species identified using the classical methods. Archaeological hair often lacks analyzable hair scale patterns in microscopic analyses and polymer chain reaction (PCR)-based techniques are often inapplicable due to the lack of amplifiable ancient DNA. To overcome these drawbacks, a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method was used exclusively based on hair keratins. Thirteen fur specimens from his accoutrement were analyzed after tryptic digest of native hair. Peptide mass fingerprints (pmfs) from ancient samples and from reference species mostly occurring in the Alpine surroundings at his lifetime were compared to each other using multidimensional scaling and binary hierarchical cluster tree analysis. Both statistical methods highly reflect spectral similarities among pmfs as close zoological relationships. While multidimensional scaling was useful to discriminate specimens on the zoological order level, binary hierarchical cluster tree reached the family or subfamily level. Additionally, the presence and/or absence of order, family and/or species-specific diagnostic masses in their pmfs allowed the identification of mammals mostly down to single species level. Red deer was found in his shoe vamp, goat in the leggings, cattle in his shoe sole and at his quivers closing flap as well as sheep and chamois in his coat. Canid species, like grey wolf, domestic dog or European red fox, were discovered in his leggings for the first time, but could not be differentiated to species level. This is widening the spectrum of processed fur-bearing species to at least one member of the Canidae family. His fur cap was allocated to a carnivore species, but differentiation between brown bear and a canid species could not be made with certainty.

Genetic variation across the current range of the Asian houbara bustard (Chlamydotis undulata macqueenii)

The houbara bustard (Chlamydotisundulata), the main quarry for Arab falconry,is currently threatened by excessive huntingand poaching as well as by habitat loss andfragmentation. We have investigated the geneticdiversity, population structure and demographichistory of houbara bustards across theirgeographical range by analyzing mitochondrial(mt)DNA sequences (a 370 bp fragment of controlregion I and a 264 bp fragment of thecytochrome b gene) and 4 microsatelliteloci in 74 individuals sampled from the CanaryIslands to China. Both markers revealed low tomoderate diversity that could be partitionedinto two monophyletic groups or evolutionarysignificant units belonging to the NorthAfrican (C. u. undulata) and Asian (C. u. macqueenii) subspecies. A history ofrelatively recent population growth (∼35,000years ago) accompanied by range expansion isthe most likely demographic scenario for theAsian subspecies. In addition, Macqueensbustards are able to disperse efficiently overbroad areas, which is consistent with ourinference of weak phylogeographic structure(global FST = 0.20 and 0.04 formtDNA and microsatellites, respectively) andhigh levels of homogenizing gene flow on widegeographical scales. We therefore suggest thatmanagement actions should focus on maintainingmigratory connectivity between breeding andnon-breeding areas.

DNA-led rediscovery of the giant sable antelope in Angola

The giant sable antelope (Hippotragus niger variani), unique to Angola, was feared extinct after almost three decades of civil war. Comparisons of mitochondrial DNA sequences derived from dung samples recently collected in the field and from old museum specimens of certain provenance provide the first documented evidence to date that this enigmatic antelope has survived. Its DNA-led rediscovery in the former combat zone was subsequently confirmed by photographic evidence. The Angolan isolate constitutes a distinct monophyletic group that shows a dramatic population decline from historic levels. It represents a diagnosable conservation unit which is characterised by unique cranial morphological features (Blaine 1922 1922:317–339), a highly restricted range, and the presence of fixed genetic differences in all of its common relatives.