Liselotte Wesley Andersen

Habitat fragmentation causes bottlenecks and inbreeding in the European tree frog (Hyla arborea).

A genetic study of the European tree frog, Hyla arborea, in Denmark was undertaken to examine the population structure on mainland Jutland and the island of Lolland after a period of reduction in suitable habitat and population sizes. The two regions have experienced the same rate of habitat loss but fragmentation has been more severe on Lolland. Genetic variation based on 12 polymorphic DNA microsatellites was analysed in 494 tree frogs sampled from two ponds in Jutland and 10 ponds on Lolland. A significant overall deviation from Hardy–Weinberg expectations could be attributed to three ponds, all on Lolland. This was most probably caused by an inbreeding effect reducing fitness, which was supported by the observed significant negative correlation between larva survival and mean FIS value and mean individual inbreeding coefficient. A significant reduction in genetic variation (bottleneck) was detected in most of the ponds on Lolland. Population–structure analysis suggested the existence of at least 11 genetically different populations, corresponding to most of the sampled population units. The results indicated that the populations were unique genetic units and could be used to illustrate the migration pattern between newly established ponds arisen either by natural colonization of tree frogs or by artificial introduction. A high degree of pond fidelity in the tree frogs was suggested. A severe fragmentation process reducing population size and fitness within some of the populations probably caused the significant reduction in genetic variation of tree frog populations on Lolland.

Population structure and gene flow of the Atlantic walrus (Odobenus rosmarus rosmarus) in the eastern Atlantic Arctic based on mitochondrial DNA and microsatellite variation

The population structure of the Atlantic walrus, Odobenus rosmarus rosmarus, was studied using 11 polymorphic microsatellites and restriction fragment length polymorphism detected in the NADH‐dehydrogenase ND1, ND2 and ND3/4 segments in mtDNA. A total of 105 walrus samples were analysed from northwest (NW) Greenland, east (E) Greenland, Svalbard and Franz Joseph Land. Two of the 10 haplotypes detected in the four samples were diagnostic for the NW Greenland sample, which implied that the group of walruses in this area is evolutionary distinct from walruses in the other three areas. One individual sampled in E Greenland exhibited a Pacific haplotype, which proved a connection between the Pacific walrus and walruses in eastern Greenland. The Franz Joseph Land, Svalbard and E Greenland samples shared the most common haplotype, indicating very little differentiation at the mtDNA level. Gene flow (Nm) estimates among the four areas indicated a very restricted exchange of female genes between NW Greenland and the more eastern Atlantic Arctic samples, and a closer relationship between the three samples composing the eastern Atlantic Arctic. The genetic variation at 11 polymorphic microsatellite loci grouped individuals into three populations, NW Greenland, E Greenland and a common Franz Joseph Land–Svalbard population, which were connected by moderate gene flow.

Conservation genetics of harbour porpoises, Phocoena phocoena, in eastern and central North Atlantic

We examined polymorphism at 12 microsatelliteloci in 807 harbour porpoises , Phocoenaphocoena, collected from throughout thecentral and eastern North Atlantic to theBaltic Sea. Multilocus tests for allelefrequency differences, assignment tests,population structure estimates (FST) andgenetic distance measures (DLR andDC) all indicate six geneticallydifferentiated populations/sub-populationsafter pooling sub-samples within regions.Harbour porpoises from West Greenland, theNorwegian Westcoast, Ireland, the British NorthSea, the Danish North Sea and the inland watersof Denmark (IDW) are all geneticallydistinguishable from each other. A sample ofharbour porpoises collected off the Dutch coast(mainly during winter) was geneticallyheterogeneous and likely comprised a mixture ofindividuals of diverse origin. A mixed stockanalysis indicated that most of the individualsin this sample (∼77%) were likely migrantsfrom the British and Danish North Sea.

Mitochondrial Control Region and microsatellite analyses on harbour porpoise (Phocoena phocoena) unravel population differentiation in the Baltic Sea and adjacent waters

The population status of the harbour porpoise (Phocoena phocoena) in the Baltic area has been a continuous matter of debate. Here we present the by far most comprehensive genetic population structure assessment to date for this region, both with regard to geographic coverage and sample size: 497 porpoise samples from North Sea, Skagerrak, Kattegat, Belt Sea, and Inner Baltic Sea were sequenced at the mitochondrial Control Region and 305 of these specimens were typed at 15 polymorphic microsatellite loci. Samples were stratified according to sample type (stranding vs. by-caught), sex, and season (breeding vs. non-breeding season). Our data provide ample evidence for a population split between the Skagerrak and the Belt Sea, with a transition zone in the Kattegat area. Among other measures, this was particularly visible in significant frequency shifts of the most abundant mitochondrial haplotypes. A particular haplotype almost absent in the North Sea was the most abundant in Belt Sea and Inner Baltic Sea. Microsatellites yielded a similar pattern (i.e., turnover in occurrence of clusters identified by STRUCTURE). Moreover, a highly significant association between microsatellite assignment and unlinked mitochondrial haplotypes further indicates a split between North Sea and Baltic porpoises. For the Inner Baltic Sea, we consistently recovered a small, but significant separation from the Belt Sea population. Despite recent arguments that separation should exceed a predefined threshold before populations shall be managed separately, we argue in favour of precautionary acknowledging the Inner Baltic porpoises as a separate management unit, which should receive particular attention, as it is threatened by various factors, in particular local fishery measures.

A combined DNA-microsatellite and isozyme analysis of the population structure of the harbour porpoise in Danish waters and West Greenland

One hundred and twenty-four specimens of the harbour porpoise, Phocoena phocoena, occurring in inner Danish waters (IDW), the North Sea and West Greenland were analysed to study subdivision into genetically differentiated subpopulations using PCR-amplified DNA-microsa-tellites and isozyme markers. Three polymorphic microsatellites, 415/416, 417/418 and Igf-I (insulin-like growth factor I) were detected with nine, eight and 15 alleles, respectively, and from a former study two polymorphic isozymes, Mpi-1 and Pgm, with three and two alleles, respectively, were used in the analysis. Overall deviations from the expected Hardy-Weinberg distribution were only observed in the total sample and at a single locus in the North Sea-summer sample and at two loci in the West Greenland sample. Whenever this occurred a surplus of homozygotes was observed, suggesting a Wahlund effect, a null allele or nonrandom mating. The analysis of the genetical population structure showed that harbour porpoises from West Greenland, the North Sea and IDW were three geographically, genetically differentiated populations even though connected through some degree of gene flow. A tendency for females to be more stationary than males was suggested. Furthermore, the population structure suggested a closer relationship between IDW and the North Sea.

Integrating genetic data and population viability analyses for the identification of harbour seal (Phoca vitulina) populations and management units

Identification of populations and management units is an essential step in the study of natural systems. Still, there is limited consensus regarding how to define populations and management units, and whether genetic methods allow for inference at the relevant spatial and temporal scale. Here, we present a novel approach, integrating genetic, life history and demographic data to identify populations and management units in southern Scandinavian harbour seals. First, 15 microsatellite markers and model‐ and distance‐based genetic clustering methods were used to determine the population genetic structure in harbour seals. Second, we used harbour seal demographic and life history data to conduct population viability analyses (PVAs) in the vortex simulation model in order to determine whether the inferred genetic units could be classified as management units according to Lowe and Allendorfs (Molecular Ecology, 19, 2010, 3038) ‘population viability criterion’ for demographic independence. The genetic analyses revealed fine‐scale population structuring in southern Scandinavian harbour seals and pointed to the existence of several genetic units. The PVAs indicated that the census population size of each of these genetic units was sufficiently large for long‐term population viability, and hence that the units could be classified as demographically independent management units. Our study suggests that population genetic inference can offer the same degree of temporal and spatial resolution as ‘nongenetic’ methods and that the combined use of genetic data and PVAs constitutes a promising approach for delineating populations and management units.

The population structure of the harbour porpoise, Phocoena phocoena, in Danish waters and part of the North Atlantic

A study of the homogeneity between putative stocks of Phocoena phocoena in the North Sea and inner Danish waters was performed using isozyme electrophoresis. Two polymorphic systems, mannose phosphate isomerase (MPI) and phosphoglucomutase (PGM) were used in the analysis. A two-locus homogeneity test showed an effect of season and locality on the genotypic distribution. This led to a division of the total sample into season and into the two localities, the inner Danish waters (IDW) and the North Sea. The samples were then compared to the Hardy-Weinberg expectations where a deficit in heterozygotes was observed in the North Sea sample in both systems and at the Pgm locus in the total sample, indicating a mixing of sub-populations. A hierarchical contingency table analysis, also based on the genotypic distribution, showed a combined effect of season and locality and only an insignificant effect of sampling period, thus supporting the division into season and locatily. The seasonal effect is further supported by earlier observations of a seasonal migration of harbour porpoises out of the Baltic in winter. A significant difference was observed between the two supposed sub-populations, IDW-summer and North Sea-summer samples, based on the genotypic distribution. This could also be a reflection of the sample sizes. On the basis of samples obtained from Canada, West Greenland and Holland a preliminary study of the population structure on a larger scale, comprising the North East and North West Atlantic, was performed. The results so far accord well with the hitherto accepted sub-populations of P. phocoena in the North Atlantic.

A review of the genetic relationships of Atlantic walrus (Odobenus rosmarus rosmarus) east and west of Greenland

Abstract. Studies of the genetic variation involving allozymes, mitochondrial and nuclear DNA (microsatellites) in walruses (Odobenus rosmarus) were reviewed. In addition, the genetic relationships of a total of 211 Atlantic walruses, O. r. rosmarus, from 5 sampling areas west and east of Greenland were studied using 12 nuclear DNA-microsatellite loci and restriction fragment length polymorphism obtained from the ND1, ND2 and ND3/4 segments of the mitochondrial DNA (mtDNA). At the mtDNA level, no divergence was observed among the three sampling areas east of Greenland (i.e. East Greenland, Svalbard and Franz Josef Land), whereas areas west of Greenland (i.e. Northwest and West Greenland) showed some differentiation. The genetic variation at the microsatellite loci grouped the individuals into four sub-populations: Northwest Greenland, West Greenland, East Greenland and a common Svalbard-Franz Josef Land sub-population. A significant correlation between genetic distance and geographic distance between the sampling areas (isolation-by-distance effect) was detected, especially at the mtDNA level. At a small-scale phylo-geographical level, the mtDNA data indicated that Atlantic walruses have diverged into two major groups: one northwest (i.e. in the North Water) and one east of Greenland (i.e. an East Greenland-Svalbard-Franz Josef Land group), whereas the haplotype distribution in the West Greenland sample reflected a mixture of both these groups. The microsatellite data supported a general grouping of walruses to the west and east of Greenland.

The Laptev Sea walrus Odobenus rosmarus laptevi: an enigma revisited

The walrus (Odobenus rosmarus) is in some current systematic schemes divided into three subspecies: O. r. rosmarus in the North Atlantic, O. r. divergens in the North Pacific and O. r. laptevi in the Laptev Sea. These three subspecies have been described as differing in body size, but the taxonomic status of O. r. laptevi is disputed. The current study applies molecular and morphometric methods to assess the taxonomic status of O. r. laptevi and to analyse the systematic and phylogeographic relationships between the three purported walrus subspecies. Tusk length and tusk circumference were measured from the few skulls available of O. r. laptevi, and the obtained values were within the ranges reported for Pacific walruses. Thus, morphologically, subspecies status for O. r. laptevi is not supported according to the Amadon–Mayr ‘75% rule’. Phylogenetic analyses and haplotype networks based on mitochondrial nucleotide sequence data of NADH dehydrogenase 1, 16S rRNA, cytochrome oxidase I and the d‐loop of the control region of the historic O. r. laptevi bone material and contemporary O. r. rosmarus and O. r. divergens showed that the Laptev Sea walrus groups with individuals from the North Pacific. Thus, the mitochondrial sequence data do not support the recognition of three walrus subspecies as reciprocally monophyletic evolutionary units with independent evolutionary histories. Only O. r. rosmarus and O. r. divergens meet this criterion with the present sampling. Accordingly, we recommend that Odobenus r. laptevi be abandoned and the Laptev walrus instead be recognized as the westernmost population of the Pacific walrus, Odobenus r. divergens. However, further research is recommended to assess whether the Laptev walrus could be considered as a significant unit in terms of conservation and management, since it is unique in several ecological parameters.

Harbour porpoises ( Phocoena phocoena ) in the North Atlantic: Distribution and genetic population structure

The known geographical distribution (based on ship surveys, aerial surveys, incidental sightings, stranding and bycatch data) and the population genetic structure obtained from mitochondria DNA and nuclear DNA (isozymes and microsatellites) data analyses of the harbour porpoise in the North Atlantic have recently been reviewed and revised by the International Whaling Commission. The present review builds on these documents by integrating more recent genetic and distributional studies. Studies of the genetic structure of harbour porpoise populations tend to be concentrated in areas where samples are available which coincide with areas where incidental or directed catches or stranding take place. Nevertheless, recently, several genetic studies on the population structure have been able to reveal a more comprehensive picture of the harbour porpoise population structure in the Northwest and Northeast Atlantic, although not all areas have been subjected to analyses.