The Genetic Integrity of the Ex Situ Population of the European Wildcat (Felis silvestris silvestris) Is Seriously Threatened by Introgression from Domestic Cats (Felis silvestris catus)

Studies on the genetic diversity and relatedness of zoo populations are crucial for implementing successful breeding programmes. The European wildcat, Felis s. silvestris, is subject to intensive conservation measures, including captive breeding and reintroduction. We here present the first systematic genetic analysis of the captive population of Felis s. silvestris in comparison with a natural wild population. We used microsatellites and mtDNA sequencing to assess genetic diversity, structure and integrity of the ex situ population. Our results show that the ex situ population of the European wildcat is highly structured and that it has a higher genetic diversity than the studied wild population. Some genetic clusters matched the breeding lines of certain zoos or groups of zoos that often exchanged individuals. Two mitochondrial haplotype groups were detected in the in situ populations, one of which was closely related to the most common haplotype found in domestic cats, suggesting past introgression in the wild. Although native haplotypes were also found in the captive population, the majority (68%) of captive individuals shared a common mtDNA haplotype with the domestic cat (Felis s. catus). Only six captive individuals (7.7%) were assigned as wildcats in the STRUCTURE analysis (at K = 2), two of which had domestic cat mtDNA haplotypes and only two captive individuals were assigned as purebred wildcats by NewHybrids. These results suggest that the high genetic diversity of the captive population has been caused by admixture with domestic cats. Therefore, the captive population cannot be recommended for further breeding and reintroduction.     

High genetic diversity in the blue-listed British Columbia population of the purple martin maintained by multiple sources of immigrants

To assess genetic diversity in the blue-listed purple martin (Progne subis) population in British Columbia, we analysed mitochondrial control region sequences of 93 individuals from British Columbia and 121 individuals collected from seven localities of the western and eastern North American subspecies P. s. arboricola and P. s. subis, respectively. Of the 47 haplotypes we detected, 34 were found exclusively in western populations, and 12 were found only in eastern populations. The most common eastern haplotype (25) was also found in three nestlings in British Columbia and one in Washington. Another British Columbia nestling had a haplotype (35) that differed by a C to T transition from haplotype 25. Coalescent analysis indicated that these five nestlings are probably descendents of recent immigrants dispersing from east to the west, because populations were estimated to have diverged about 200,000–400,000 ybp, making ancestral polymorphism a less likely explanation. Maximum likelihood estimates of gene flow among all populations detected asymmetrical gene flow into British Columbia not only of rare migrants from the eastern subspecies in Alberta but also a substantial number of migrants from the adjacent Washington population, and progressively lower numbers from Oregon in an isolation-by distance pattern. The influx of migrants from different populations is consistent with the migrant-pool model of recolonization which has maintained high genetic diversity in the small recovering population in British Columbia. Thus, the risk to this population is not from genetic erosion or inbreeding following a severe population crash, but from demographic stochasticity and extinction in small populations.     

Genetic diversity in the US ex situ populations of the endangered Puerto Rican Boa,Chilabothrus inornatus

The Puerto Rican Boa (Chilabothrus inornatus) was placed on the US Endangered Species List in 1970. Progress has been made since to clarify the recovery status of this species, though the design of a new recovery plan must include information regarding genetic variation within and among populations of this species. While measures of genetic diversity in wild populations of this species are finally becoming available, relative genetic diversity represented in ex situ populations is unknown, which hampers efforts to develop an ex situ species management plan. Here, we provide an analysis of genetic diversity in US public and private collections (zoos and breeders) using mitochondrial sequence data and five highly polymorphic nuclear microsatellite loci. We analyzed 50 boas from the US ex situ population and determined overall genetic diversity and relatedness among these individuals. We then compared these data to mitochondrial and microsatellite data obtained from 176 individuals from wild populations across the native range of the species. We found little inbreeding and a large amount of retained genetic diversity in the US ex situ population of C. inornatus relative to wild populations. Genetic diversity in the ex situ population is similar to that found in wild populations. Ours is only the second explicit attempt to characterize genetic diversity at the molecular level in ex situ populations of boid snakes. We anticipate that these results will inform current breeding strategies as well as offer additional information that will facilitate the continuation of ex situ conservation breeding or management in boas.     

Gene survival in the Asian wild horse (Equus przewalskii): II. Gene survival in the whole population,in subgroups,and through history

In populations with a known pedigree, exact joint probability distributions of numbers of surviving of genes from each founder can now be calculated for moderately large complex pedigrees (1,000–2,000 individuals and much inbreeding). The usefulness of such calculations is shown by our analysis of gene survival in the Asian wild horse (Equus przewalskii), a species now extinet in the wild with a captive population with 1,516 individuals in the known pedigree (12 generations). We calculate the genetic diversity of subsets of the current population interesting to the North American Species Survival Plan, trace the loss of genetic diversity in this species through its history in captivity, and determine genetically important individuals in the North American population—those with relatively high probabilities of having unique copy genes (genes not found in any other living individual in North America).     

In situ population structure and ex situ representation of the endangered Amur tiger

The Amur tiger ( Panthera tigris altaica ) is a critically endangered felid that suffered a severe demographic contraction in the 1940s. In this study, we sampled 95 individuals collected throughout their native range to investigate questions relative to population genetic structure and demographic history. Additionally, we sampled targeted individuals from the North American ex situ population to assess the genetic representation found in captivity. Population genetic and Bayesian structure analyses clearly identified two populations separated by a development corridor in Russia. Despite their well-documented 20th century decline, we failed to find evidence of a recent population bottleneck, although genetic signatures of a historical contraction were detected. This disparity in signal may be due to several reasons, including historical paucity in population genetic variation associated with postglacial colonization and potential gene flow from a now extirpated Chinese population. Despite conflicting signatures of a bottleneck, our estimates of effective population size ( N _e = 27–35) and N _e /N ratio (0.07–0.054) were substantially lower than the only other values reported for a wild tiger population. Lastly, the extent and distribution of genetic variation in captive and wild populations were similar, yet gene variants persisted ex situ that were lost in situ . Overall, our results indicate the need to secure ecological connectivity between the two Russian populations to minimize loss of genetic diversity and overall susceptibility to stochastic events, and support a previous study suggesting that the captive population may be a reservoir of gene variants lost in situ .     

Characterizing the molecular variation among American marten (Martes americana) subspecies from Oregon and California

We investigated the subspecific identity of a rediscovered population of American martens within the range of a presumed extinct subspecies (Martes americana humboldtensis) by comparing mitochondrial DNA sequence diversity from contemporary individuals within the described ranges of M. a. humboldtensis, nearby ranges of M. a. caurina and M. a. sierrae, and a museum specimen of M. a. humboldtensis. Martens from the rediscovered population shared a haplotype (#2) with the museum specimen. This haplotype was found only in the coastal regions of Oregon and California, suggesting that the rediscovered population represents descendants of a relictual population that previously existed in coastal California. The subspecific boundary between M. a. humboldtensis and M. a. caurina may not be valid, because haplotype #2 was shared between coastal Oregon and coastal California populations and no known contemporary or historical biogeographic barriers prevent north–south movement. Thus, marten populations currently located in coastal forests of California and Oregon should be managed collectively to preserve the connectivity that our data suggest occurred historically. M. a. sierrae differed substantially from both M. a. humboldtensis and M. a. caurina, suggesting marten populations were not a historically genetically homogeneous population and divergence may have occurred in separate glacial refugia.     

Phylogeography and conservation genetics of Eld's deer (Cervus eldi)

Eld's deer (Cervus eldi) is a highly endangered cervid, distributed historically throughout much of South Asia and Indochina. We analysed variation in the mitochondrial DNA (mtDNA) control region for representatives of all three Eld's deer subspecies to gain a better understanding of the genetic population structure and evolutionary history of this species. A phylogeny of mtDNA haplotypes indicates that the critically endangered and ecologically divergent C. eldi eldi is related more closely to C. e. thamin than to C. e. siamensis, a result that is consistent with biogeographic considerations. The results also suggest a strong degree of phylogeographic structure both between subspecies and among populations within subspecies, suggesting that dispersal of individuals between populations has been very limited historically. Haplotype diversity was relatively high for two of the three subspecies (thamin and siamensis), indicating that recent population declines have not yet substantially eroded genetic diversity. In contrast, we found no haplotype variation within C. eldi eldi or the Hainan Island population of C. eldi siamensis, two populations which are known to have suffered severe population bottlenecks. We also compared levels of haplotype and nucleotide diversity in an unmanaged captive population, a managed captive population and a relatively healthy wild population. Diversity indices were higher in the latter two, suggesting the efficacy of well-designed breeding programmes for maintaining genetic diversity in captivity. Based on significant genetic differentiation among Eld's deer subspecies, we recommend the continued management of this species in three distinct evolutionarily significant units (ESUs). Where possible, it may be advisable to translocate individuals between isolated populations within a subspecies to maintain levels of genetic variation in remaining Eld's deer populations.     

Comparative molecular studies on the genetic diversity of an ex situ garden collection and its source population of the critically endangered polish endemic plant <Emphasis Type="Italic">Cochlearia polonica</Emphasis> E. Fröhlich

Cochlearia polonica (Brassicaceae) is a narrow endemic plant extinct in the wild, known only from one transplanted population in southern Poland. The suitable habitats for this species are confined to shallow water courses and springs on sandy ground. During the second half of twentieth century the natural populations of C. polonica declined due to anthropogenic effects (primarily due to a major change of ground water levels). Consequently, 14 individuals were transplanted in the 1970 s into a secondary locality of similar habitat conditions of Centuria river. In the late 1980s five individuals from the transplanted population were used to establish an artificial ex situ site in the Botanical Garden of the Polish Academy of Sciences in Warsaw. In order to investigate the effect of 18 years of ex situ conservation, the level of genetic diversity and genetic structure of the ex situ conserved botanical garden population and its source population were sampled and analyzed using inter simple sequence repeat markers. The percentage values of polymorphic bands and Nei’s gene diversity indicated average genetic variation at species level, whereas at the population level it was relatively low, especially for the garden population. The results suggested differences in genetic composition of the analyzed populations and implied homogenization of the genetic structure of the ex situ conserved population. Ex situ conservation resulted in a decrease of the species’ genetic diversity, implying that the artificial population only partly represents the primary genetic variability found in the source population.     

Colonization and divergence: phylogeography and population genetics of the Atlantic coast beach mice

Barrier island taxa provide an opportunity to investigate recent evolutionary processes, such as colonization and isolation of recently diverged taxa, and provide important insights into understanding contemporary diversity and the assessment of conservation units. Using rapidly evolving genetic markers (mitochondrial DNA and microsatellites), we studied the Atlantic coast beach mouse subspecies (Peromyscus polionotus decoloratus, P. p. niveiventris, and P. p. phasma). Our data indicate that each of the extant coastal subspecies (P. p. niveiventris and P. p. phasma) is comprised of unique haplotypes indicative of their isolation, while the extinct subspecies, P. p. decoloratus, contain a single haplotype, which was shared with P. p. phasma. Moreover, all the coastal haplotypes originate from a single mainland haplotype found in central Florida, USA. The microsatellite data indicated high levels of genetic structure among our sampled populations. Additionally, these data group the populations into three distinct genetic clusters, with each of the extant coastal subspecies belonging to their own cluster and the mainland individuals forming a separate cluster. The extant Atlantic coast beach mice are on separate evolutionary trajectories, thus representative of separate taxonomic units. Therefore, the data support that two extant subspecies on the Florida Atlantic coast fit the Distinct Population Segment designation and should be managed and conserved as two separate independent units.     

RECENT VERSUS RELIC: DISCERNING THE GENETIC SIGNATURE OF FUCUS VESICULOSUS (HETEROKONTOPHYTA; PHAEOPHYCEAE) IN THE NORTHWESTERN ATLANTIC1

Fucus vesiculosus L. is one of the most widespread macrophytes in the northwestern Atlantic, ranging from North Carolina (USA) to Greenland (DK). We investigated genetic diversity, population differentiation, patterns of isolation by distance, and putative glacial refugial populations across seven locations from North Carolina (USA) to Cape Breton Island, Nova Scotia (Canada), with microsatellite analyses. Distinct northern versus southern (Delaware–North Carolina) populations were revealed by microsatellite data. Five of six microsatellite loci were fixed in populations in North Carolina, suggesting a recent founder event or a bottleneck, and the same homozygous genotype was found in herbarium materials collected on the North Carolina coast from more than 60 years ago. An additional set of individuals from the northern limit in Greenland was included in our analysis of mitochondrial intergenic spacer (mt IGS) haplotypes in the northwestern Atlantic. Remarkably, 184 of 188 F. vesiculosus specimens from North Carolina to Greenland shared the same haplotype. Recent colonization of the North American shore from Europe is hypothesized based upon the ubiquity of this common haplotype, which was earlier reported from Europe.     

Isolation of 18 polymorphic microsatellite loci from the North American red squirrel,Tamiasciurus hudsonicus (Sciuridae,Rodentia), and their cross‐utility in other species

We isolated 18 polymorphic microsatellite loci to be used for pedigree analysis in a wild population of North American red squirrels, Tamiasciurus hudsonicus. Allelic diversity and observed heterozygosity ranged from six to 13 and 0.39 to 0.89, respectively, in a sample of 93 individuals. Up to 13 sets of primers also amplify in other rodent species.     

Genetic diversity in wild and reared populations of the Japanese bitterling <Emphasis Type="Italic">Tanakia tanago</Emphasis> (Cyprinidae)

 The Japanese bitterling Tanakia tanago is an endangered cyprinid species; thus, captive breeding programs are being conducted in various facilities as ex situ conservation. To examine the genetic diversity in one wild and three reared populations, and its changes during the process of captive breeding, sequences of the mitochondrial cytochrome b gene and control region were determined. The wild population, collected in 1993, was monomorphic. Although the reared population that originated from the wild population was almost monomorphic, a rare haplotype, distinct from all others by a relatively large sequence divergence, was also observed. In the other reared populations, some degree of genetic diversity had been maintained. A reared hybrid population, which originated from a mixture of three distinct populations, showed the greatest genetic diversity. These results suggest considerable genetic diversity within and among populations of T. tanago in the past. Although a loss of genetic diversity was observed in some year-classes of reared populations, there was no tendency for genetic diversity to decrease as a result of captive breeding, probably because offspring were obtained from multi-year-class parents in the captive breeding program. Accordingly, this breeding method should be appropriate for conserving the genetic diversity of T. tanago. Received: June 12, 2002 / Revised: December 3, 2002 / Accepted: December 16, 2002     

The cryptic genetic structure of the North American captive gorilla population

Western lowland gorillas (Gorilla gorilla gorilla) were imported from across their geographical range to North American zoos from the late 1800s through 1974. The majority of these gorillas were imported with little or no information regarding their original provenance and no information on their genetic relatedness. Here, we analyze 32 microsatellite loci in 144 individuals using a Bayesian clustering method to delineate clusters of individuals among a sample of founders of the captive North American zoo gorilla collection. We infer that the majority of North American zoo founders sampled are distributed into two distinct clusters, and that some individuals are of admixed ancestry. This new information regarding the existence of ancestral genetic population structure in the North American zoo population lays the groundwork for enhanced efforts to conserve the evolutionary units of the western lowland gorilla gene pool. Our data also show that the genetic diversity estimates in the founder population were comparable to those in wild gorilla populations (Mondika and Cross River), and that pairwise relatedness among the founders is no different from that expected for a random mating population. However, the relatively high level of relatedness (R = 0.54) we discovered in a pair of known breeding pairs reveals the need for incorporating genetic relatedness estimates in the captive management of western lowland gorillas.     

天鹅洲迁地保护江豚群体的遗传评估与发展预测

文章以2008年春季遭受严重雪灾的天鹅洲白鱀豚国家级自然保护区迁地保护江豚群体为研究对象,利用物种特异性微卫星遗传标记对该群体的遗传多样性进行了评估,并通过软件对其发展趋势进行了模拟和预测。研究结果表明,虽然2008年春季雪灾并未导致该群体遗传多样性的严重损失,灾后群体依然表现出中等水平的微卫星遗传多样性(18个座位的平均等位基因数为4.780,平均观察杂合度和期望杂合度分别为0.748和0.674);但模拟结果显示,该迁地保护江豚群体的遗传多样性将快速下降,100年后仅能保留约79%的等位基因和约84%的遗传杂合度。此外,模拟结果表明,通过增加群体数量和调整群体性比可有效减缓该群体遗传多样性的下降速度。根据研究结果,并结合长江江豚自然种群快速下降的现状,作者提出天鹅洲迁地保护江豚群体当前最重要的保护策略应该是尽早补充一定数量的、合适的野生个体,在加强遗传多样性保护的同时使其群体数量快速增长并形成一个更大规模的有效繁殖群体。     

Assessing changes in genomic divergence following a century of human‐mediated secondary contact among wild and captive‐bred ducks

Along with manipulating habitat, the direct release of domesticated individuals into the wild is a practice used worldwide to augment wildlife populations. We test between possible outcomes of human‐mediated secondary contact using genomic techniques at both historical and contemporary timescales for two iconic duck species. First, we sequence several thousand ddRAD‐seq loci for contemporary mallards (Anas platyrhynchos) throughout North America and two domestic mallard types (i.e., known game‐farm mallards and feral Khaki Campbell's). We show that North American mallards may well be becoming a hybrid swarm due to interbreeding with domesticated game‐farm mallards released for hunting. Next, to attain a historical perspective, we applied a bait‐capture array targeting thousands of loci in century‐old (1842–1915) and contemporary (2009–2010) mallard and American black duck (Anas rubripes) specimens. We conclude that American black ducks and mallards have always been closely related, with a divergence time of ~600,000 years before present, and likely evolved through prolonged isolation followed by limited bouts of gene flow (i.e., secondary contact). They continue to maintain genetic separation, a finding that overturns decades of prior research and speculation suggesting the genetic extinction of the American black duck due to contemporary interbreeding with mallards. Thus, despite having high rates of hybridization, actual gene flow is limited between mallards and American black ducks. Conversely, our historical and contemporary data confirm that the intensive stocking of game‐farm mallards during the last ~100 years has fundamentally changed the genetic integrity of North America's wild mallard population, especially in the east. It thus becomes of great interest to ask whether the iconic North American mallard is declining in the wild due to introgression of maladaptive traits from domesticated forms. Moreover, we hypothesize that differential gene flow from domestic game‐farm mallards into the wild mallard population may explain the overall temporal increase in differentiation between wild black ducks and mallards, as well as the uncoupling of genetic diversity and effective population size estimates across time in our results. Finally, our findings highlight how genomic methods can recover complex population histories by capturing DNA preserved in traditional museum specimens.     

Genetic assessment of the Iberian wolf Canis lupus signatus captive breeding program

The main goal of ex situ conservation programs is to improve the chances of long term survival of natural populations by founding and managing captive colonies that can serve as a source of individuals for future reintroductions or to reinforce existing populations. The degree in which a captive breeding program has captured the genetic diversity existing in the source wild population has seldom been evaluated. In this study we evaluate the genetic diversity in wild and captive populations of the Iberian wolf, Canis lupus signatus, in order to assess how much genetic diversity is being preserved in the ongoing ex situ conservation program for this subspecies. A sample of domestic dogs was also included in the analysis for comparison. Seventy-four wolves and 135 dogs were genotyped at 13 unlinked microsatellite loci. The results show that genetic diversity in Iberian wolves is comparable in magnitude to that of other wild populations of gray wolf. Both the wild and the captive Iberian wolf populations have a similarly high genetic diversity indicating that no substantial loss of diversity has occurred in the captive-breeding program. The effective number of founders of the program was estimated as ∼ ∼16, suggesting that all founders in the studbook pedigree were genetically independent. Our results emphasize also the genetic divergence between wolves and domestic dogs and indicate that our set of 13 microsatellite loci provide a powerful diagnostic test to distinguish wolves, dogs and their hybrids.     

Limited genetic diversity of an endemic subspecies Schizopygopsis chengi baoxingensis as inferred from the mitochondrial DNA control region

As a subspecies of Schizopygopsis chengi (Fang), Schizopygopsis chengi baoxingensis (Fu, Ding et Ye) occurs as a single population endemic to the Baoxing River in China. The influences from the construction of hydropower dams in its distribution area might result in its extinction in the future due to its very narrow habitat and small population size. In this study, we sequenced the mitochondrial DNA control region of 81 individuals to analyze the genetic variability of the subspecies, hoping to acquire basic genetic information. The results revealed a limited genetic diversity of the population. Total samples could only be defined as seven haplotypes based on nucleotide variations. The nucleotide diversity (0.0008), haplotype diversity (0.534), and overall mean genetic distance among individuals (0.00080 ± 0.00053) were very low. Therefore, it is very necessary and urgent to conserve the population of this subspecies, avoiding further genetic loss, or even extinction. Handling editor: Christian Sturmbauer     

Subspecies of the Central American Squirrel Monkey (Saimiri oerstedii) as Units for Conservation

The accurate diagnosis of conservation units now typically includes recognition of genetic diversity and unique evolutionary lineages and is necessary to inform the conservation management of endangered species. We evaluated whether the two currently recognized subspecies of the endangered Central American squirrel monkey (Saimiri oerstedii) in Costa Rica are evolutionarily significant units (ESUs) that should be managed separately in conservation efforts. We used previously published sequences of 50 individuals of Saimiri oerstedii for 880 bp of the mtDNA d-loop and genotypes of 244 individuals for 16 microsatellites and conducted novel analyses to characterize genetic differentiation between subspecies of Saimiri oerstedii. We measured sequence differentiation and inferred an intraspecific molecular phylogeny and a haplotype network, and found consistent results supporting statistically significant divergence and reciprocal monophyly between subspecies. A population aggregation analysis also supported Saimiri oerstedii citrinellus and S. o. oerstedii as diagnosably distinct units. These results confirm previous genetic studies with smaller sample sizes and are consistent with other factors including differences in pelage and morphology and divergence at nuclear markers. Conservation managers should manage these subspecies separately to prevent the loss of genetic diversity via artificially induced outbreeding. High levels of genetic diversity may buffer populations against outside extinction pressures, to which Saimiri oerstedii are vulnerable because of their dwindling habitat and small population size.     

Genetic variation and population structure in remnant populations of black rhinoceros, Diceros bicornis, in Africa

Black rhinoceros (Diceros bicornis) are one of the most endangered mammal species in Africa, with a population decline of more than 96% by the end of the last century. Habitat destruction and encroachment has resulted in fragmentation of the remaining populations. To assist in conservation management, baseline information is provided here on relative genetic diversity and population differentiation among the four remaining recognized subspecies. Using microsatellite data from nine loci and 121 black rhinoceros individuals, and comparing the results with those of other African species affected in similar ways, Diceros bicornis michaeli retained the most genetic diversity (heterozygosity 0.675) compared with Diceros bicornis minor (0.459) and Diceros bicornis bicornis (0.505), suggesting that the duration of the known bottlenecks in these populations has only had a limited impact on diversity. Comparable and moderate degrees of population differentiation were found between D. b. minor, D. b. bicornis and D. b. michaeli. Results from the single sample available of the most endangered subspecies, Diceros bicornis longipes, showed the least diversity of all individuals examined. This information should assist conservation management decisions, especially those affecting population viability assessments and selection of individuals for translocations, and will also facilitate subspecies identification for ex situ individuals of uncertain origin.     

Lost populations and preserving genetic diversity in the lion Panthera leo: Implications for its ex situ conservation

Two of the eight recognized lion subspecies, North African Barbary lion (Panthera leo leo) and South African Cape lion (Panthera leo melanochaita), have become extinct in the wild in the last 150 years. Based on sequences of mitochondrial DNA (mtDNA) control region (HVR1) extracted from museum specimens of four Barbary and one Cape lion, the former was probably a distinct population characterized by an invariable, unique mtDNA haplotype, whilst the latter was likely a part of the extant southern African lion population. Extinction of the Barbary line, which may still be found in “generic” zoo lions, would further erode lion genetic diversity. Therefore, appropriate management of such animals is important for maintaining the overall genetic diversity of the species. The mtDNA haplotype unique to the Barbary lion, in combination with the small size of the HVR1 analyzed (c. 130 bp), makes it possible and cost-effective to identify unlabelled Barbary specimens kept in museums and “generic” captive lions that may carry the Barbary line. An initial study of five samples from the lion collection of the King of Morocco, tested using this method, shows that they are not maternally Barbary.