Primeval conditions – what can we learn from them?

In this paper I argue that prehistoric British forests and their avifauna were similar to those persisting in a near-primeval condition in the Białowieża Forest (Polish/Belarussian border). Therefore, observations in the Białowieża Forest may serve as a baseline against which effects of changes in the British woods can be assessed. The results of long-term (30 years) studies of the breeding birds in the Białowieża National Park are summarized. They reveal that the local avifauna shows some features (high species richness, low population densities, high nest predation and low production of young) regarded as typical for undisturbed tropical forests. It is proposed that high productivity, high population densities and sedentary habits found in many British populations are to a large extent a recent phenomenon, due to forest fragmentation, reduction of predator diversity and changes in forest structure. The necessity to preserve the whole Białowieża Forest – currently seriously threatened by ongoing logging – as a benchmark for future biological studies is underlined.     

Analysis of molecular differentiation in a hybrid zone between chromosomally distinct races of the common shrew Sorex araneus (Insectivora: Soricidae) suggests their common ancestry

During postglacial colonization, populations that diverged in different refugia produced a patchwork of genomes, often delimited with sharp hybrid zones. The outcome of hybridization following the secondary contact of two genetically distinct populations is hard to predict. In this context, the present study investigated the genetic structure of the hybrid zone between the Drnholec and Białowieża chromosome races of the common shrew ( Sorex araneus ) in Poland using biparentally inherited (seven autosomal microsatellites) and uniparentally inherited (Y-linked microsatellite and mtDNA) molecular markers. On the basis of diagnostic chromosomes, the Drnholec and Białowieża races were classified to different karyotypic groups, which were believed to have independent glacial histories. It was found that genetic differentiation between the Drnholec and Białowieża races was weak and nonsignificant with respect to all molecular markers. However, these results are in contrast with the chromosomal structure of this hybrid zone. The very sharp frequency clines of the diagnostic chromosomes strongly suggest that gene flow between the Drnholec and Białowieża races was reduced. Nonsignificant correlations between genetic differentiation and both the presence of an environmental barrier and geographical distance reveal that only differences in karyotypes might be a reason for limited gene exchange between the races. It is assumed that a lack of molecular differences between the Drnholec and Białowieża races results from a shared ancestral variation.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 79–90.     

Population ecology and conservation of endangered megafauna: the case of European bison in Białowieża Primeval Forest, Poland

The European bison Bison bonasus is an example of nearly extinct 'charismatic megafauna'. The Białowieża Primeval Forest in Poland is among the few places where they still live in the wild. The management of this free-living herd has to reconcile to the conservation needs of a species and the economic and environmental objectives of their habitat: protected as well as commercial woodlands of Białowieża. Here we present a detailed account of the population development and analyse variation in vital rates based on monitoring that started in 1952 and continued until 2002. The population was allowed to grow freely until 1970, when removal started with the aim to stabilize population size. We found that recruitment rate, but not mortality, was density dependent, suggesting that the population density was not very high relative to resource levels. Winters with much snow and cold temperature had a strong negative effect on survival. May temperature of the previous year positively affected recruitment rates. Masting (oak seed) also positively affected recruitment rates, which provides a rare account of masting affecting the performance of a large ruminant. Sex ratio of offspring was even and was not strongly affected by density or climate. We use an age-structured matrix model to show how this knowledge of intrinsic and extrinsic factors affecting vital rates may help managerial decisions by providing explicit links between given environmental conditions and the population growth rates.     

Body mass and measurements of the European bison during postnatal development

Body mass of the European bisonBison bonasus (Linnaeus, 1758) was determined for 152 animals (86 males, 66 females) from captive breeding centre in Białowieża and for 492 (248, 244) animals from the free-ranging herd in the Białowieża Forest. Body measurements were taken for a total of 472 bison (204, 268). Age-related changes in body mass and measurements of 375 bison (165, 210) were analysed. Bison body mass was significantly correlated with age. Captive bison were heavier than those from a free-ranging herd. Measurements showed the changes in bison body structure in the postnatal development. All measurements were highly correlated with age and mass of the animals and increased most intensively during the first year of life. Sex-related differences in body mass and measurements were most pronounced in bison ≥ 3 years old. Physical development of females ends at the age of 5 years, and that of males at the age of 7 years.     

MHC-DRB3 variation in a free-living population of the European bison, Bison bonasus

MHC genes play a crucial role in pathogen recognition and are the most polymorphic genes in vertebrates. Loss of variation in these genes in bottlenecked species is thought to put their survival at risk. We examined variation at the MHC II DRB3 locus in the European bison, Bison bonasus, a species that has undergone an extreme bottleneck: the current population originated from only 12 founders. We also tested for the association of DRB3 genes with the incidence of posthitis, a disease affecting the reproductive organs of bulls and posing a new threat to the survival of the species. We found very limited MHC diversity, with only four alleles segregating in a sample of 172 individuals from a free‐ranging Białowieża population. The alleles were highly divergent and revealed the hallmark of positive selection acting on them in the past, that is, a significant excess of nonsynonymous substitutions. This excess was concentrated in putative antigen‐binding sites, suggesting that selection was driven by pathogens. However, we did not observe departures from Hardy–Weinberg equilibrium, an indicator of strong ongoing selection. Neither have we found a significant association between DRB3 alleles or genotypes and susceptibility to posthitis. Alleles conferring resistance to males may have been lost during the extreme bottleneck the species had undergone.     

Genetic variability in the European bison (Bison bonasus) population from Białowieża forest over 50 years

The aim of this study was to assess potential post-bottleneck temporal genetic differentiation following the reintroduction of the species into the Białowieża Forest. Variability of 12 polymorphic microsatellite markers was analysed for 178 individuals born between 1955 and 2005, divided by birth year into five temporal groups. Low overall allelic richness (AR) per locus (AR = 2.0) and a low overall expected heterozygosity (H_E = 0.28) were observed. The overall F _IS was not significantly different from zero. The mean F _IS values were, however, significantly different from zero for individuals born between 1955 and 1965 ( F _IS = 0.19). A Bayesian computation was used to estimate effective population size (N_e) for each temporal group. We observed relatively small values of N_e ranging from 7.9 to 28.4. The low N_e values confirm that, despite a rapid growth of the bison population following the founder event, N_e increased only slowly.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 801–809.     

Detection of mitochondrial DNA from domestic cattle in bison on Santa Catalina Island

In 1924, 14 American bison (Bison bison) were introduced to Santa Catalina Island, California and sporadically supplemented thereafter with additional animals. To reduce the herd and its impact on native vegetation, over 2000 animals have been exported during the past four decades. Today, the herd is estimated to contain around 250 individuals. Genetic analysis was performed on 98 animals removed from the island in 2004. Forty-four samples (45%) had domestic cattle mitochondrial DNA (mtDNA), 12 (12%) had previously reported bison haplotypes and 42 (43%) had a new haplotype differing by one base pair from a previously reported bison haplotype. A complement of five restriction enzymes was found to be useful in identifying bison with domestic cattle mtDNA.     

Differential introgression of uniparentally inherited markers in bison populations with hybrid ancestries

Historical hybridization between Bison bison (bison) and Bos taurus (cattle) has been well documented and resulted in cattle mitochondrial DNA (mtDNA) introgression, previously identified in six different bison populations. In order to examine Y chromosome introgression, a microsatellite marker (BYM-1) with non-overlapping allele size distributions in bison and cattle was isolated from a bacterial artificial chromosome (BAC) clone, and was physically assigned to the Y chromosome by fluorescence in situ hybridization. BYM-1 genotypes for a sample of 143 male bison from 10 populations, including all six populations where cattle mtDNA haplotypes were previously identified, indicated that cattle Y chromosome introgression had not occurred in these bison populations. The differential permeability of uniparentally inherited markers to introgression is consistent with observations of sterility among first generation hybrid males and a sexual asymmetry in the direction of hybridization favouring matings between male bison and female cattle.     

Genetic status of the European bison Bison bonasus after extinction in the wild and subsequent recovery

• 1 The European bison Bison bonasus went through a severe bottleneck and became extinct in the wild 90 years ago. The lowland subspecies B. b. bonasus is the only one of three original subspecies that exists today. The entire species derives from only 12 founders, including a bull of the Caucasian subspecies B. b. caucasicus. Due to its presence among founders, there are two geographically separated genetic lines of European bison: the pure lowland (Bia?owie?a) line and the hybrid lowland‐Caucasian line.
• 2 The lowland line of the European bison originates from only seven founders with an extremely varying genetic contribution. Approximately 80% of the genes in contemporary populations come from just two founders.
• 3 A variety of genetic markers (mtDNA, microsatellites, single nucleotide polymorphism microchips) were applied to studies of the level of depletion of genetic variability in European bison.
• 4 The lowland line of the European bison, the most extensively studied, shows very low levels of genetic variation, and has just half the microsatellite heterozygosity of the closely related American bison Bison bison. The effective population size (N_e) for the highly genetically homogenous lowland line in the Polish part of the Bia?owie?a Forest is estimated to be 23.5, far less than the census population size of 450.
• 5 The average inbreeding level in lowland bison is almost 50%, although no signs of inbreeding depression have been observed. In contrast, inbreeding effects have been noticed in the lowland‐Caucasian line, which has a much lower average inbreeding level (28%). In spite of the apparently high fitness of the lowland bison, the lack of genetic variation and high level of inbreeding may present substantial threats in the future, especially in the context of potential epizootics.

     

Mother-calf relationships and maternal investment in European bison<Emphasis Type="Italic">Bison bonasus</Emphasis>

Mother-young relationships, up to the third month of the calves’ life, were investigated in European bisonBison bonasus (Linnaeus, 1758) from Białowieża National Park breeding centres (Poland). During the first week, calves spent most of their time close to their mothers and were rarely seen in the vicinity of other group members. Highest suckling rate, most frequent vocal contact and agonistic behaviour of mothers towards intruders, were other characteristics of this period. Although being a forest species, European bison exhibit a following type of strategy for offspring protection, typical for ungulates living in open areas; no hiding phase was observed. Suckling rates were similar in calves of both sexes, however, the pattern of nursing was different: male calves suckled longer, but in rarer bouts than female calves. The mothers’ condition, reflected by their social rank, did not influence the suckling rate. No sex differences were recorded in other maternal activities or mother-calf distance. The presumption of a lack of sex-biased maternal investment in European bison is confirmed by the results of other research.     

The strength and timing of the mitochondrial bottleneck in salmon suggests a conserved mechanism in vertebrates

In most species mitochondrial DNA (mtDNA) is inherited maternally in an apparently clonal fashion, although how this is achieved remains uncertain. Population genetic studies show not only that individuals can harbor more than one type of mtDNA (heteroplasmy) but that heteroplasmy is common and widespread across a diversity of taxa. Females harboring a mixture of mtDNAs may transmit varying proportions of each mtDNA type (haplotype) to their offspring. However, mtDNA variants are also observed to segregate rapidly between generations despite the high mtDNA copy number in the oocyte, which suggests a genetic bottleneck acts during mtDNA transmission. Understanding the size and timing of this bottleneck is important for interpreting population genetic relationships and for predicting the inheritance of mtDNA based disease, but despite its importance the underlying mechanisms remain unclear. Empirical studies, restricted to mice, have shown that the mtDNA bottleneck could act either at embryogenesis, oogenesis or both. To investigate whether the size and timing of the mitochondrial bottleneck is conserved between distant vertebrates, we measured the genetic variance in mtDNA heteroplasmy at three developmental stages (female, ova and fry) in chinook salmon and applied a new mathematical model to estimate the number of segregating units (N(e)) of the mitochondrial bottleneck between each stage. Using these data we estimate values for mtDNA Ne of 88.3 for oogenesis, and 80.3 for embryogenesis. Our results confirm the presence of a mitochondrial bottleneck in fish, and show that segregation of mtDNA variation is effectively complete by the end of oogenesis. Considering the extensive differences in reproductive physiology between fish and mammals, our results suggest the mechanism underlying the mtDNA bottleneck is conserved in these distant vertebrates both in terms of it magnitude and timing. This finding may lead to improvements in our understanding of mitochondrial disorders and population interpretations using mtDNA data.     

High ectoparasite loads in hole‐nesting birds – a nestbox bias?

All studies of interactions between fleas and hole-nesting birds so far have been done in nestboxes. This study, which is the first study of flea infestation of nests of hole-nesters in natural holes, was carried out in the Białowieża National Park (E Poland) and demonstrated that, in contrast to those studies, both prevalence (below 10%) and intensity of infestation nests of Parus palustris and Ficedula flycatchers were very low, an order of magnitude lower than elsewhere. However, in the same forest, flycatchers breeding in nestboxes, and tits in natural holes within the nestbox plot, had significantly higher infestation rates, approaching those found in other studies. These observations indicate that the high flea loads reported for tits and flycatchers may be a product of biased sampling – exclusive collection of data in nestbox areas – rather than reflect their species-specific characteristics. The data from natural holes undermine the widespread assumption that data on flea loads in nestboxes can be treated as representative, and generalised to areas without nestboxes.     

Stable heteroplasmy at the single-cell level is facilitated by intercellular exchange of mtDNA

Eukaryotic cells carry two genomes, nuclear (nDNA) and mitochondrial (mtDNA), which are ostensibly decoupled in their replication, segregation and inheritance. It is increasingly appreciated that heteroplasmy, the occurrence of multiple mtDNA haplotypes in a cell, plays an important biological role, but its features are not well understood. Accurately determining the diversity of mtDNA has been difficult, due to the relatively small amount of mtDNA in each cell (<1% of the total DNA), the intercellular variability of mtDNA content and mtDNA pseudogenes (Numts) in nDNA. To understand the nature of heteroplasmy, we developed Mseek, a novel technique to purify and sequence mtDNA. Mseek yields high purity (>90%) mtDNA and its ability to detect rare variants is limited only by sequencing depth, providing unprecedented sensitivity and specificity. Using Mseek, we confirmed the ubiquity of heteroplasmy by analyzing mtDNA from a diverse set of cell lines and human samples. Applying Mseek to colonies derived from single cells, we find heteroplasmy is stably maintained in individual daughter cells over multiple cell divisions. We hypothesized that the stability of heteroplasmy could be facilitated by intercellular exchange of mtDNA. We explicitly demonstrate this exchange by co-culturing cell lines with distinct mtDNA haplotypes. Our results shed new light on the maintenance of heteroplasmy and provide a novel platform to investigate features of heteroplasmy in normal and diseased states.     

Heteroplasmic point mutations in the human mtDNA control region.

As part of an investigation of the fixation mechanisms of mtDNA mutations in humans, we sequenced the first hypervariable segment of the control region in 180 twin pairs and found evidence of site heteroplasmy in 4 pairs. Significant levels of two mitochondrial haplotypes differing by a single point mutation were found in two MZ pairs, and within each pair, both members had similar levels of heteroplasmy. Two DZ pairs were found in which the predominant mitochondrial haplotype differed within the pair. We measured proportions of mitochondrial haplotypes within two twin pairs and their maternal relatives, using primer extension. In both maternal lineages, most family members were heteroplasmic, and the proportions of each genotype varied widely in different individuals. We used the changes in haplotype proportions within mother-offspring pairs to calculate the size range of potential bottlenecks in mitochondrial numbers occurring during development of the offspring. In most individuals, the most likely effective bottleneck sizes ranged from 3 to 20 segregating units, though in two individuals a small bottleneck was very unlikely and there was no upper limit on its possible size. We also used the data from this study, together with unpublished data from other populations, to estimate the frequency of site heteroplasmy in normal human populations. From this, we calculated that the rate of mutation and fixation in the first hypervariable segment of the human mtDNA control region is between 1.2 x 10(-6) and 2.7 x 10(-5) per site per generation. This range is in good agreement with published estimates calculated by other methods.     

Application of mtDNA d-loop region for the study of Russian sturgeon population structure from Iranian coastline of the Caspian Sea

The population structure of the Russian sturgeon ( Acipenser gueldenstaedtii ) was investigated using PCR amplification of the mtDNA D-loop region. Seven composite haplotypes were detected and average nucleotide and haplotype diversity over all samples were found to be 0.05 ± 0.00 and 0.75 ± 0.00 (mean ± SE) respectively. Restriction digest of the mtDNA D-loop region detected two genotypes A and B with a relative high frequencies of 0.5 and 0.36 respectively. These two genotypes (A and B) can be considered as potential genetic markers for different biological groups, stocks or seasonal races of Russian sturgeon. A maximum of 10.8% nucleotide diversity was found between haplotypes AAAAB and BBBBF. 100% heteroplasmy was found in Russian sturgeon and restriction digest of the mtDNA D-loop region also exhibited site heteroplasmy.     

Space Use and Movement Patterns in a Semi-Free-Ranging Herd of European Bison (Bison bonasus)

The successful reintroduction and restocking of the European Bison demands a reliable knowledge of the biology of this species. Yet little is known to date about the European bison, and empirical data remains insufficient to set up a reliable plan ensuring the reintroduction, maintenance and survival of populations in habitats that have been largely modified by human activity. Studies of the ecology, social behaviour and management of bison are therefore crucial to the conservation of this species and its cohabitation with humans. To meet these challenges, we focused on movement patterns and space use in a semi-free-ranging herd of European bison living in the Réserve Biologique des Monts-d’Azur (France). Bison spend over 80% of their time foraging and resting; foraging mainly occurs around the artificial feeding sites (i.e., hay racks) or in meadows. The time of day and the presence of snow have no influence on the time budget allocated to each activity. Animals, however, spend more time at the food racks in winter. Bison also spend most of their time in small groups of individuals, confirming the occurrence of both fission-fusion dynamics and sexual segregation in this species. Bison seem to follow a Lévy walk pattern of movement, which is probably related to the geographical distribution and size of food patches in the reserve. The conclusions of this study provide a better understanding of the sociality, life habits and habitat use of bison, and also describe how the provision of hay affects all these behaviours. These results could be useful in the development of tools to select the most suitable habitats for the reintroduction, management and conservation of bison populations.     

Morphometric variation of the skull during postnatal development in the Lowland European bisonBison bonasus bonasus

We studied the variation of linear measurements and skull capacity in Lowland European bisonBison bonasus bonasus (Linnaeus, 1758) during postnatal development, and the dependencies of the parameters in relation to sex, age, and body mass of the animals. Material consisted of 599 bison skulls (310 males and 289 females), within the age range of 1 month to 21 years (males) and to 27 years (females). In the group of calves to 1 year old, no sex connected differences in skull measurements were observed, whereas the skull capacity in older calves was significantly larger (0.01>p>0.001) in males than in females. From the third year of life, most skull measurements display characteristics of sexual dimorphism. Skull development in both sexes is most intensive during the first three years of life, and slows from the age of 5. In older individuals of both sexes (≥ 6 years), orbital breadth continues growing and, in females, breadth of splanchnocranium continues increasing. Growth in a bison’s skull capacity is most intensive up to the third year of life and slows from the age of 5. During postnatal development, a bison skull grows proportionally except the neurocranium, which grows slightly slower in comparison with basal length and its development finishes earlier than that of splanchnocranium. In ontogenesis, a bison skull grows much slower compared to body mass. In relation to body mass, skull capacity and the height of neurocranium grow most slowly while orbital breadth grows most intensively. The results obtained were compared with data on skull sizes of bison born in 1930–1950 and bred in captivity and with skulls of the American bisonBison bison. Inbreeding is probably responsible for some types of phenotypic abnormalities in the skull which appear in modern European bison.     

Effects of a recent founding event and intrinsic population dynamics on genetic diversity in an ungulate population

Maintenance of genetic diversity has recently become a management goal for a number of species, due to its importance for present and future population viability. Genetic drift, primarily through differential reproductive success and inbreeding, can accelerate the loss of genetic diversity in recently recovered populations. We attempt to quantify the consequences of these factors on the genetic diversity contained in a small, recently founded wood bison (Bison bison athabascae) population by examining the genetic variation in this conservation herd, the calves born therein, and its large source population. The Hook Lake Wood Bison Recovery Project was initiated to found a disease-free herd of wood bison containing a representative amount of the genetic diversity present in the Wood Buffalo National Park metapopulation. Levels of diversity in the Hook Lake Wood Bison Recovery Project founders are higher than in previous salvage attempts. To examine the effects of differential reproductive success on this population, we monitored parentage of the calves born in the Hook Lake Wood Bison Recovery Project for 3 years since the founders reached sexual maturity. Two of the male founders sired over 90% of the offspring born in this population, which has led to a reduction in diversity in their calves. Monitoring of reproductive success, and incorporation of selective breeding strategies will be required to reduce the rate at which genetic diversity is lost from this small, isolated population. These steps should occur in other recovery projects, particularly when a small number of individuals are capable of dominating reproduction.     

Genetic variability in the European bison

In the European bison, Bison bonasus (L), 15 proteins coded by 20 loci have been studied. Two of these loci (Es-3 and Ca) are polymorphic and the remaining 18 are monomorphic. The average heterozygosity of bisons in Polish herds is 3.5%. The value does not differ from those found in populations of other large mammals in spite of restoration of the European bison population from a mere 13 individuals.     

Mitochondrial and nuclear DNA analyses reveal pronounced genetic structuring in Tunisian wild boar <Emphasis Type="Italic">Sus scrofa</Emphasis>

We analysed 74 wild boars from Tunisia with respect to patterns of genetic differentiation and diversity based on sequences of the mitochondrial control region and genotypes at eight nuclear microsatellite loci. Analysis of molecular variance for both marker systems and Bayesian structure analysis of our microsatellite data revealed a clear break between northern and southern populations. Southern wild boar were monomorphic for one of three mtDNA haplotypes; the other two (one of which only occurred in three individuals) were confined to the north. A comparison with published sequences showed all three haplotypes to belong to the major European clade E1. Microsatellite diversity was similar to that found in earlier studies of wild boar (expected heterozygosity of 0.695 and 0.597 for the north and south, respectively). Contrary to the mtDNA results, we did not find unequivocal evidence of a bottleneck in Tunisian wild boar based on our microsatellite data. The clear distinction between northern and southern populations may be due to an Algerian origin of the southern animals.