ANALYSIS OF MECHANISMS OF MICROEVOLUTIONARY CHANGE IN CEPPHUS GUILLEMOTS USING PATTERNS OF CONTROL REGION VARIATION

We surveyed population-level sequence variation in part of the mitochondrial control region for three species including eight subspecies of Cepphus guillemots (Charadriiformes: Alcidae) to test specific predictions about mechanisms of population differentiation. We found that sequences of spectacled guillemots (C. carbo) were more closely related to those of pigeon guillemots (C. columba; both found in the Pacific Ocean) than to those of black guillemots (C. grylle; Arctic and Atlantic Oceans), despite dissimilarities in plumage between spectacled guillemots and the other species. Distributions of species and timing of divergence events suggest that speciation involved allopatric and microallopatric populations isolated by Pleistocene glaciers. Control region sequences were significantly differentiated among populations within species and suggest that gene flow is low; however, populations are probably not in genetic equilibrium, so these results probably reflect historical isolation of colonies. In contrast, phylogenetic relationships among sequences within species were poorly resolved, probably because of a combination of incomplete lineage sorting and contemporary gene flow. Indices of genetic diversity provided no suggestion of recent bottlenecks in most populations, although two populations apparently underwent recent severe bottlenecks. Genetic divergence among populations was not correlated with geographic distance, which argues against isolation by distance. Results of these analyses, combined with breeding distributions and timing of divergence events, suggest that populations diverged during isolation in glacial refugia. Our results are consistent with earlier hypotheses posed by Storer and Udvardy.     

Molecular population genetic analysis of the spectacled bear (Tremarctos ornatus) in the northern Andean area

Eighty-two Andean bear samples obtained in three South American Andean countries were analyzed using five hypervariable microsatellite markers. Neither the overall sample nor the samples coming from each of the countries analyzed were in Hardy-Weinberg equilibrium. This is attributable to the Wahlund effect caused by a fragmentation of the bear populations. The genetic variability levels found were in general low for this kind of molecular markers (H = 0.38 for the global population). The situation of the Ecuadorian bear population seems to be endangered (H = 0.24). This is the lowest level recorded for any bear population. The genetic heterogeneity among the three populations was large and highly significant (F(ST) = 0.39; R(ST) = 0.32-0.49). Likewise, the gene flow estimates were remarkably low among these populations (Nm = 0.2-0.3). This means that the populations are strongly isolated. Different methods were applied to determine the effective numbers in these populations. A significant spatial structure of the genotypes was analyzed with different procedures. In all cases, an isolation-by-distance structure was detected. This could be a consequence of the original South America colonization. There was no evidence for a recent genetic bottleneck. This could mean that the low heterozygosity and the population fragmentation could be explained by ancient events related to the bear colonization, with the arrival of humans beings in the Americas, and/or with the glacial maximum, 16.000-30.000 years ago.     

Genetic consequences of habitat fragmentation and loss: the case of the Florida black bear (<Emphasis Type="Italic">Ursus americanus floridanus</Emphasis>)

Habitat loss and fragmentation can influence the genetic structure of biological populations. We studied the genetic consequences of habitat fragmentation in Florida black bear (Ursus americanus floridanus) populations. Genetic samples were collected from 339 bears, representing nine populations. Bears were genotyped for 12 microsatellite loci to estimate genetic variation and to characterize genetic structure. None of the nine study populations deviated from Hardy–Weinberg equilibrium. Genetic variation, quantified by mean expected heterozygosity (H _E), ranged from 0.27 to 0.71 and was substantially lower in smaller and less connected populations. High levels of genetic differentiation among populations (global F _ST = 0.224; global R _ST = 0.245) suggest that fragmentation of once contiguous habitat has resulted in genetically distinct populations. There was no isolation-by-distance relationship among Florida black bear populations, likely because of barriers to gene flow created by habitat fragmentation and other anthropogenic disturbances. These factors resulted in genetic differentiation among populations, even those that were geographically close. Population assignment tests indicated that most individuals were genetically assigned to the population where they were sampled. Habitat fragmentation and anthropogenic barriers to movement appear to have limited the dispersal capabilities of the Florida black bear, thereby reducing gene flow among populations. Regional corridors or translocation of bears may be needed to restore historical levels of genetic variation. Our results suggest that management actions to mitigate genetic consequences of habitat fragmentation are needed to ensure long-term persistence of the Florida black bear.     

Phylogeny of the bears (Ursidae) based on nuclear and mitochondrial genes

The taxomic classification and phylogenetic relationships within the bear family remain argumentative subjects in recent years. Prior investigation has been concentrated on the application of different mitochondrial (mt) sequence data, herein we employ two nuclear single-copy gene segments, the partial exon 1 from gene encoding interphotoreceptor retinoid binding protein (IRBP) and the complete intron 1 from transthyretin (TTR) gene, in conjunction with previously published mt data, to clarify these enigmatic problems. The combined analyses of nuclear IRBP and TTR datasets not only corroborated prior hypotheses, positioning the spectacled bear most basally and grouping the brown and polar bear together but also provided new insights into the bear phylogeny, suggesting the sister-taxa association of sloth bear and sun bear with strong support. Analyses based on combination of nuclear and mt genes differed from nuclear analysis in recognizing the sloth bears as the earliest diverging species among the subfamily ursine representatives while the exact placement of the sun bear did not resolved. Asiatic and American black bears clustered as sister group in all analyses with moderate levels of bootstrap support and high posterior probabilities. Comparisons between the nuclear and mtDNA findings suggested that our combined nuclear dataset have the resolving power comparable to mtDNA dataset for the phylogenetic interpretation of the bear family. As can be seen from present study, the unanimous phylogeny for this recently derived family was still not produced and additional independent genetic markers were in need.     

Population genetic structure of two water strider species in the Ecuadorian Amazon

1. We used mtDNA sequence variation to estimate population genetic structure between and among water strider populations of Potamobates williamsi and P . sumaco in the Ecuadorian Amazon.
2. Sequencing of the COI mitochondrial gene revealed 16 haplotypes, which were summarised into four haplotype groups. P . williamsi and P . sumaco shared two common and widespread haplotypes.
3. Population structure was moderate and gene flow was low. Both air and river distances were significantly correlated with gene flow and, thus, indicative of isolation by distance.
4. The genetic structure within and among populations of P. williamsi and P. sumaco was probably not influenced by the dynamic tropical lotic system.     

Relationships among cave swallow populations (Petrochelidon fulva) determined by comparisons of microsatellite and cytochrome b data

The cave swallow (Petrochelidon fulva) is a polytypic species with isolated populations in northwestern South America, southwestern North America, Yucatan, Greater Antilles, and Florida. We compared microsatellite genotypes of at least five individuals each from five populations and cytochrome b sequence data of two individuals each from seven populations plus two outgroups. Microsatellite allelic diversity was substantial, and the North American populations were about equally variable. In contrast, the Ecuadorian population had far less genetic variation. Gene flow was apparent among populations, especially between Texas and Florida. Genetic structure was greater than in widespread North American species but less than that of sedentary Neotropical birds. Microsatellite genetic distances indicated a close affinity between Ecuadorian and northern populations, especially Texas and Florida, but this finding was inconsistent with cytochrome b data, which indicated that the Ecuadorian population is the clear outgroup to the northern populations. Its outgroup status is consistent with recent classifications that designate South American populations as their own species (P. rufocollaris). The cytochrome b data further suggested that the northern populations are divided into two clades: Texas/Yucatan and Florida/Greater Antilles. The microsatellite data incorrectly measured the diversity and affinities of Ecuadorian birds apparently because of an ascertainment bias that results from the use of heterologous PCR primers. Despite these problems in measuring phylogenetic relationships, the microsatellite data appeared to work well as a population genetic marker in indicating population structure and gene flow.     

GENETIC VARIATION IN TIME AND SPACE: MICROSATELLITE ANALYSIS OF EXTINCT AND EXTANT POPULATIONS OF ATLANTIC SALMON

Information on genetic composition of past and present populations may be obtained by analyzing DNA from archival samples. A study is presented on the genetic population structure of extant and extinct local populations of Atlantic salmon from 1913 to 1989 using dried scales as a source of DNA. Variation at six microsatellite loci was studied. Tests for differentiation among populations and among time series within populations showed that population structure was stable over time. This was also confirmed by a neighbor-joining dendrogram, which showed a clear clustering of samples from individual rivers that covered a time span of up to 76 years. These results suggest that salmon populations evolve as semi-independent units connected by modest amounts of gene flow. Additionally, a clear association between geographic and genetic distance was found. This relationship has otherwise been difficult to establish in several recent studies. The discrepancy may be due to impact of human activities on the genetic structure of present populations, whereas old samples represent populations in a more unaffected state. However, other explanations related to differences in the sampling of past and present populations may be equally valid.     

Genetic diversity and differentiation of Kermode bear populations

The Kermode bear is a white phase of the North American black bear that occurs in low to moderate frequency on British Columbia's mid-coast. To investigate the genetic uniqueness of populations containing the white phase, and to ascertain levels of gene flow among populations, we surveyed 10 highly polymorphic microsatellite loci, assayed from trapped bear hairs. A total of 216 unique bear genotypes, 18 of which were white, was sampled among 12 localities. Island populations, where Kermodes are most frequent, show approximately 4% less diversity than mainland populations, and the island richest in white bears (Gribbell) exhibited substantial genetic isolation, with a mean pairwise FST of 0.14 with other localities. Among all localities, FST for the molecular variant underlying the coat-colour difference (A893G) was 0.223, which falls into the 95th percentile of the distribution of FST values among microsatellite alleles, suggestive of greater differentiation for coat colour than expected under neutrality. Control-region sequences confirm that Kermode bears are part of a coastal or western lineage of black bears whose existence predates the Wisconsin glaciation, but microsatellite variation gave no evidence of past population expansion. We conclude that Kermodism was established and is maintained in populations by a combination of genetic isolation and somewhat reduced population sizes in insular habitat, with the possible contribution of selective pressure and/or nonrandom mating.     

Nuclear DNA microsatellite analysis of genetic diversity and gene flow in the Scandinavian brown bear (Ursus arctos)

In the 1930s, the Scandinavian brown bear was close to extinction due to vigorous extermination programmes in Norway and Sweden. Increased protection of the brown bear in Scandinavia has resulted in the recovery of four subpopulations, which currently contain close to 1000 individuals. Effective conservation and management of the Scandinavian brown bear requires knowledge of the current levels of genetic diversity and gene flow among the four subpopulations. Earlier studies of mitochondrial DNA (mtDNA) diversity revealed extremely low levels of genetic variation, and population structure that grouped the three northern subpopulations in one genetic clade and the southernmost subpopulation in a second highly divergent clade. In this study, we extended the analysis of genetic diversity and gene flow in the Scandinavian brown bear using data from 19 nuclear DNA microsatellite loci. Results from the nuclear loci were strikingly different than the mtDNA results. Genetic diversity levels in the four subpopulations were equivalent to diversity levels in nonbottlenecked populations from North America, and significantly higher than levels in other bottlenecked and isolated brown bear populations. Gene flow levels between subpopulations ranged from low to moderate and were correlated with geographical distance. The substantial difference in results obtained using mtDNA and nuclear DNA markers stresses the importance of collecting data from both types of genetic markers before interpreting data and making recommendations for the conservation and management of natural populations. Based on the results from the mtDNA and nuclear DNA data sets, we propose one evolutionarily significant unit and four management units for the brown bear in Scandinavia.     

Low genetic diversities in isolated populations of the Asian black bear (<Emphasis Type="Italic">Ursus thibetanus</Emphasis>) in Japan,in comparison with large stable populations

Populations of the Asian black bear (Ursus thibetanus) are relatively large and continuous in central Honshu, the main island of Japan, but they are isolated in western Honshu. To clarify the degree of genetic isolation of the populations in western Honshu, we compared the genetic diversities of four populations in western Honshu with that of one of the continuous populations of central Honshu. Three of the four western Honshu populations were isolated and the other was continuous with the central Honshu populations on a geographical distribution basis. The genotypes at 10 microsatellite loci of the sampled individuals were determined and the genetic structures of the populations examined. Genetic diversities were significantly lower in the isolated populations than in the continuous populations. The continuous population in central Honshu had high levels of genetic diversity, comparable to those in populations of the American black bear (Ursus americanus) and the brown bear (Ursus arctos). The genetic distances between the two continuous populations were smallest, even though their geographic distance was largest (>200 km) among all the pairs of neighboring populations examined. Low genetic diversity within the isolated populations suggested genetic drift due to the small population size; the genetic differentiation among the populations indicated low rates of gene flow among them.     

Using stage‐based system dynamics modeling for demographic management of captive populations

Management of captive populations relies on a complex synthesis of genetic and demographic analyses to guide populations toward sustainability. Demographic analyses of captive populations currently utilize age‐based matrix projections to predict a population's trajectory. An alternate approach is to use a stage‐based, system dynamics model for captive systems. Such models can more easily incorporate complex captive systems in which population dynamics are dependent on a combination of management and a species' biology. By linking these two areas, population managers can gain a more accurate understanding of how management decisions impact captive populations and which aspects of a species' demography should be of special concern in the future. We present a general stage‐based system dynamics model that has been developed for use with captive populations. The utility of the model is then illustrated by applying it to three captive bear populations: spectacled bears (Tremarctos ornatus), sloth bears (Melursus ursinus), and sun bears (Helarctos malayanus). Zoo Biol 22:45–64, 2003. © 2003 Wiley‐Liss, Inc.     

Phylogenetic Relationships of Bears (the Ursidae) Inferred from Mitochondrial DNA Sequences

The phylogenetic relationships among some bear species are still open questions. We present here mitochondrial DNA sequences of D-loop region, cytochrome b, 12S rRNA, tRNA^Pro, and tRNA^Thr genes from all bear species and the giant panda. A series of evolutionary trees with concordant topology has been derived based on the combined data set of all of the mitochondrial DNA sequences, which may have resolved the evolutionary relationships of all bear species: the ancestor of the spectacled bear diverged first, followed by the sloth bear; the brown bear and polar bear are sister taxa relative to the Asiatic black bear; the closest relative of the American black bear is the sun bear. Primers for forensic identification of the giant panda and bears are proposed. Analysis of these data, in combination with data from primates and antelopes, suggests that relative substitutional rates between different mitochondrial DNA regions may vary greatly among different taxa of the vertebrates.     

Ancient DNA analyses reveal high mitochondrial DNA sequence diversity and parallel morphological evolution of late pleistocene cave bears

Cave bears (Ursus spelaeus) existed in Europe and western Asiauntil the end of the last glaciation some 10,000 years ago.To investigate the genetic diversity, population history, andrelationship among different cave bear populations, we havedetermined mitochondrial DNA sequences from 12 cave bears thatrange in age from about 26,500 to at least 49,000 years andoriginate from nine caves. The samples include one individualfrom the type specimen population, as well as two small-sizedhigh-Alpine bears. The results show that about 49,000 yearsago, the mtDNA diversity among cave bears was about 1.8-foldlower than the current species-wide diversity of brown bears(Ursus arctos). However, the current brown bear mtDNA gene poolconsists of three clades, and cave bear mtDNA diversity is similarto the diversity observed within each of these clades. The resultsalso show that geographically separated populations of the high-Alpinecave bear form were polyphyletic with respect to their mtDNA.This suggests that small size may have been an ancestral traitin cave bears and that large size evolved at least twice independently.     

Genetic variation in the Heterodoxus octoseriatus group (Phthiraptera): a test of Price's model of parasite evolution.

Most of the genetic variation in the H. octoseriatus group is present as fixed gene differences between species which have been described on morphological criteria. Based on allozymes, the taxonomic status of some species was challenged. There was insufficient evidence, however, to demonstrate that these were not 'good' biological species. Overall, the limited intraspecific variation was present as fixed gene differences among lice from different hosts and from different colonies of hosts; heterozygotes were rare. Two predictions derived from Price's model of parasite evolution were met: populations of lice were genetically homogeneous and, where genetic markers were present, we found substantial genetic variation among populations. These data contrast with those for endoparasitic helminths, where, in general, the amount of genetic variation is similar to that of free-living invertebrates.     

Genetic diversity, structure, and size of an endangered brown bear population threatened by highway construction in the Pindos Mountains, Greece

One of the major negative effects of roads is the creation of barriers to the movement of wildlife, ultimately disconnecting populations and increasing extinction risk. We collected genetic data from a threatened brown bear population in the central part of the Pindos mountain range in northwestern Greece to provide information about this, as yet genetically undescribed, population and to evaluate its status prior to the construction of a major highway. We used noninvasive genetic sampling methods and microsatellite analysis to investigate nuclear genetic diversity, population genetic structure, demographic history, relatedness within the population and estimated effective and total population size. Brown bears in the study area were found to possess a relatively high level of nuclear genetic diversity and low levels of inbreeding; the population did not show any signs of substructuring but seems to have gone through a genetic bottleneck in the recent past. The estimated effective population size was 29, and the total population size estimate obtained by two different methods was 33 and 51 individuals, respectively. Our results indicate a good conservation status of this bear population and provide baseline genetic data for the future evaluation of the effects on bears from the construction of a major highway, for monitoring the genetic status of this and other bear populations in Greece and for assessing gene flow in bear populations in southern Europe.     

Serum concentrations of vitamin D metabolites, vitamins A and E, and carotenoids in six canid and four ursid species at four zoos

Nutritional status for six captive canid species (n=34) and four captive ursid species (n=18) were analyzed. The species analyzed included: African wild dog (Lycaon pictus), arctic fox (Alopex lagopus), gray wolf (Canis lupus), maned wolf (Chrysocyon brachyurus), Mexican wolf (Canis lupus baleiyi), red wolf (Canis rufus), brown bear (Ursus arctos), polar bear (Ursus maritimus), spectacled bear (Tremarctos ornatus), and sun bear (Ursus malayanus). Diet information was collected for these animals from each participating zoo (Brookfield Zoo, Fort Worth Zoo, Lincoln Park Zoological Gardens, and North Carolina Zoological Park). The nutritional composition of the diet for each species at each institution met probable dietary requirements. Blood samples were collected from each animal and analyzed for vitamin D metabolites 25(OH)D and 1,25(OH)(2)D, vitamin A (retinol, retinyl stearate, retinyl palmitate), vitamin E (alpha-tocopherol and gamma-tocopherol) and selected carotenoids. Family differences were found for 25(OH)D, retinol, retinyl stearate, retinyl palmitate and gamma-tocopherol. Species differences were found for all detectable measurements. Carotenoids were not detected in any species. The large number of animals contributing to these data, provides a substantial base for comparing the nutritional status of healthy animals and the differences among them.     

Gene flow, effective population size and selection at major histocompatibility complex genes: brown trout in the Hardanger Fjord, Norway

Brown trout populations in the Hardanger Fjord, Norway, have declined drastically due to increased exposure to salmon lice from salmonid aquaculture. We studied contemporary samples from seven populations and historical samples (1972 and 1983) from the two largest populations, one of which has declined drastically whereas the other remains stable. We analysed 11 microsatellite loci, including one tightly linked to the UBA gene of the major histocompatibility class I complex (MHC) and another locus linked to the TAP2A gene, also associated with MHC. The results revealed asymmetric gene flow from the two largest populations to the other, smaller populations. This has important conservation implications, and we predict that possible future population recoveries will be mediated primarily by the remaining large population. Tests for selection suggested diversifying selection at UBA, whereas evidence was inconclusive for TAP2A. There was no evidence for temporally fluctuating selection. We assessed the distribution of adaptive divergence among populations. The results showed the most pronounced footprints of selection between the two largest populations subject to the least immigration. We suggest that asymmetric gene flow has an important influence on adaptive divergence and constrains local adaptive responses in the smaller populations. Even though UBA alleles may not affect salmon louse resistance, the results bear evidence of adaptive divergence among populations at immune system genes. This suggests that similar genetic differences could exist at salmon louse resistance loci, thus rendering it a realistic scenario that differential population declines could reflect differences in adaptive variation.     

汩罗江与柳江鳤鱼线粒体细胞色素b基因的遗传变异初探

测定并分析了长江水系汨罗江4尾鳤鱼(Ochetobius elongates)和珠江水系柳江2尾鳤鱼的线粒体细胞色素b基因部分序列981bp,发现6个单倍型,其中19个变异位点,简约信息位点15个;AT含量和GC含量分别为57.6%和42.4%。在基于Kimura双参数模型的邻接树上,汩罗江和柳江鳤鱼各自聚群。汨罗江4尾鳤鱼间的遗传距离为0.001～0.007,柳江2尾鳤鱼间遗传距离为0.001,而2群体间的平均遗传距离为0.015(0.013～0.017),群体间的遗传变异大于群体内的遗传变异,表明汩罗江鳤鱼与柳江鳤鱼出现较明显的遗传分化。由于本研究采集地理群体少,分析的样本小,为更好地揭示长江和珠江水系鳤鱼的遗传变异,上述结论尚需进一步分析更多大样本的地理群体加以验证。     

Genetic diversity of natural Miscanthus sinensis populations in China revealed by ISSR markers

Miscanthus sinensis is a typical perennial C₄ grass in Asia. In the present study, ISSR markers were used to evaluate the genetic variation within and among grass populations that were sampled from the Zhejiang and Guangdong Provinces of China. Based on nine ISSR primers, 206 clear and reproducible DNA fragments were generated. Relatively low levels of genetic diversity were determined among the populations. The coefficient of genetic differentiation among the populations was 0.52, which indicates that 52.3% of the total molecular variance exists among the populations. Such a high level of divergence present among the populations might be caused by the complex topography and variable climatic conditions present in the two provinces. AMOVA revealed that the majority of the genetic variation was within the populations (50.6%). The application of a novel method, which combines geographical coordinates and genetic differentiation to detect barriers for gene flow, allowed us to identify two zones of lowered gene flow.     

Genetic differentiation among Danish brown trout populations, as detected RFLP analysis of PCR amplified mitochondrial DNA segments

An initial screening with 18 restriction endonucleases of brown trout mitochondrial DNA from three regions of Denmark revealed no variation in the D-loop while seven restriction endonucleases were found to detect variability in the NADH-dehydrogenase 1 and/or 5/6 regions. Thirteen different haplotypes were observed, fewer in the Lake Hald populations than in the other samples, probably a result of nonexisting gene flow due to an impassable dam. No correlation was found between geography and the genetic relationships among haplotypes. Significant genetic differentiation was observed among the three main regions. Genetic differentiation among populations was much more pronounced in the Lake Hald than in the Bornholm region. The genetic relationships among populations of the Lake Hald region suggested by mtDNA data were not in accordance with allozyme data and the known history of the populations, while estimates of genetic differentiation and gene flow inferred by the two methods were in agreement. (c) 1996 The Fisheries Society of the British Isles