Genetic diversity and population history of golden monkeys (Rhinopithecus roxellana)

Golden monkey (Rhinopithecus roxellana), namely the snub-nosed monkey, is a well-known endangered primate, which distributes only in the central part of mainland China. As an effort to understand the current genetic status as well as population history of this species, we collected a sample of 32 individuals from four different regions, which cover the major habitat of this species. Forty-four allozyme loci were surveyed in our study by allozyme electrophoresis, none of which was found to be polymorphic. The void of polymorphism compared with that of other nonhuman primates is surprising particularly considering that the current population size is many times larger than that of some other endangered species. Since many independent loci are surveyed in this study, the most plausible explanation for our observation is that the population has experienced a recent bottleneck. We used a coalescent approach to explore various scenarios of population bottleneck and concluded that the most recent bottleneck could have happened within the last 15,000 years. Moreover, the proposed simulation approach could be useful to researchers who need to analyze the non- or low-polymorphism data.     

Top-down population regulation of a top predator: lions in the Ngorongoro Crater

Efforts to determine whether bottom-up or top-down processes regulate populations have been hampered by difficulties in accurately estimating the population's carrying capacity and in directly measuring food intake rate, the impacts of interspecific competition and exposure to natural enemies. We report on 40 years of data on the lion population in Ngorongoro Crater, Tanzania, which showed strong evidence of density-dependent regulation at 100-120 individuals but has remained below 60 individuals for the past decade despite consistently high prey abundance. The lions enjoy a higher per capita food-intake rate and higher cub recruitment at low population density, and interspecific competition has not increased in recent years. These animals have suffered from a number of severe disease outbreaks over the past 40 years, but, whereas the population recovered exponentially from a severe epizootic in 1963, three outbreaks between 1994 and 2001 have occurred in such rapid succession that the population has been unable to return to the carrying capacity. The Crater population may have become unusually vulnerable to infectious disease in recent years owing to its close proximity to a growing human population and a history of close inbreeding. The Crater lions may therefore provide important insights into the future of many endangered populations.     

Fragmentation by weirs in a riverine system: A study of genetic variation in time and space among populations of European grayling (Thymallus thymallus) in a Danish river system

Human induced habitat alterations affect the genetic structure of many fish populations. Weirs in particular have caused fragmented populations previously connected by gene-flow. We studied the effects of weirs on the distribution of genetic variation within European grayling (Thymallus thymallus) populations from the Skjern River, Denmark. We compared microsatellite data from DNA extracted from historic scale samples collected 60 years ago with data from contemporary samples. Pairwise multilocus F _ST estimates between all contemporary population samples were significant as well as exact tests for population differentiation. Assignment tests of individuals to a set of baseline samples showed correct assignment to the population of origin of between 54 and 79%. Assignment of individuals from recent samples to the historic population samples showed highly variable results (3–83%) of correct assignment suggesting different population histories. Pairwise multilocus F _ST estimates were significantly correlated with the number of intervening weirs but not with waterway distance. A simulation procedure was used to estimate differences in relative population sizes, which indicated that the main river population was approximately three times larger than those of the tributaries. There were no signs of any loss of genetic variation for the river system as a whole during these 60 years. The results show that weirs can be an important factor for creating the current distribution patterns of genetic variation among grayling populations, most likely by only allowing passive downstream drift of fry and obstructing active upstream migration. This revised version was published online in July 2006 with corrections to the Cover Date.     

Unexpected Demography in the Recovery of an Endangered Primate Population

Assessments of the status of endangered species have focused on population sizes, often without knowledge of demographic and behavioral processes underlying population recovery. We analyzed demographic data from a 28-year study of a critically endangered primate, the northern muriqui, to investigate possible changes in demographic rates as this population recovered from near extirpation. As the population increased from 60 to nearly 300 individuals, its growth rate declined due to increased mortality and male-biased birth sex ratios; the increased mortality was not uniform across ages and sexes, and there has been a recent increase in mortality of prime-aged males. If not for a concurrent increase in fertility rates, the population would have stabilized at 200 individuals instead of continuing to grow. The unexpected increase in fertility rates and in adult male mortality can be attributed to the muriquis’ expansion of their habitat by spending more time on the ground. The demographic consequences of this behavioral shift must be incorporated into management tactics for this population and emphasize the importance of understanding demographic rates in the recovery of endangered species.     

A High Polymorphism Level in <Emphasis Type="Italic">Rhinopithecus roxellana</Emphasis>

The Sichuan golden monkey (Rhinopithecus roxellana) is a famous and beautiful endangered primate. Owing to the dearth of samples and technical limitations, previous studies could not adequately assess the levels of genetic polymorphisms in the species. To evaluate the polymorphisms further, we sequenced the entire mitochondrial control regions (D-loop) of 35 individuals from the 3 major habitats: Minshan, Qinglin, and Shennongjia. We observed many polymorphisms and estimated that the effective population size of the current total population is less than ca. 480–2300, and the time to the most recent common ancestor is ca. 4000–62,700 years. Through comparisons, we showed that the population of Rhinopithecus roxellana has a relatively higher polymorphism level than that of other endangered primates.     

Conservation genetics and demographic history of the endangered Cape Fear shiner (Notropis mekistocholas)

We examined allelic variation at 22 nuclear-encoded markers (21 microsatellites and one anonymous locus) and mitochondrial (mt)DNA in two geographical samples of the endangered cyprinid fish Notropis mekistocholas (Cape Fear shiner). Genetic diversity was relatively high in comparison to other endangered vertebrates, and there was no evidence of small population effects despite the low abundance reported for the species. Significant heterogeneity (following Bonferroni correction) in allele distribution at three microsatellites and in haplotype distribution in mtDNA was detected between the two localities. This heterogeneity may be due to reduced gene flow caused by a dam built in the early 1900 s. Bayesian coalescent analysis of microsatellite variation indicated that effective population size of Cape Fear shiners has declined in recent times (11-25 435 years ago, with highest posterior probabilities between 126 and 2007 years ago) by one-two orders of magnitude, consistent with the observed decline in abundance of the species. A decline in effective size was not indicated by analysis of mtDNA, where sequence polymorphism appeared to carry the signature of an older expansion phase that dated to the Pleistocene ( approximately 12 700 > 1 million years ago). Cape Fear shiners thus appear to have undergone an expansion phase following a glacial cycle but to have declined significantly in more recent times. These results suggest that rapidly evolving markers such as microsatellites may constitute a suitable tool when inferring recent demographic dynamics of populations.     

Crumbling diversity: comparison of historical archived and contemporary natural populations indicate reduced genetic diversity and increasing genetic differentiation in the golden-cheeked warbler

Genetic viability of threatened and endangered species is of increasing concern with habitat loss and fragmentation. Valuable assessments of the genetic status of endangered species are difficult in most cases, where only single sample estimates are available. Using historical and contemporary samples, we assessed the impact of both historical and recent demographic changes on population genetics of the endangered golden-cheeked warbler, (Dendroica chrysoparia). Our study documents a steep decline in genetic diversity in an endangered species over a 100-year period, along with concurrent increase in genetic differentiation, and low contemporary effective sizes for all the populations we evaluated. While adding to the growing body of literature that describes the genetic impacts of habitat fragmentation, our study may also serve as an informative guide to future management of endangered species. Our study underlines the importance of long term population genetic monitoring in understanding the full extent of genetic changes in endangered species.     

Genetic structure of an endangered raptor at individual and population levels

The Finnish population of White-tailed Eagle (Haliaeetus albicilla) has gone through two major demographic bottlenecks during the last two centuries. Strong conservation measures have allowed the population to recover, but despite the rapid population growth during recent years the species is still classified as endangered. We studied the genetic population structure at both individual and population levels in an attempt to recognize the processes shaping it. We used 9 microsatellite loci and 473 base pair fragment of the mitochondrial DNA control region on samples collected between the years 2003 and 2007 (N = 489). We found a clear isolation by distance pattern at fine scale (i.e. individual level) which is most likely a result of species’ philopatric behaviour. Although we did not find signs of the recent bottlenecks, we did find evidence of an ancient bottleneck that has occurred most likely over 21,000 years ago, long before the genetic divergence of the two present Finnish subpopulations (one along the Baltic Sea coast line and another in Lapland and easternmost Finland). We conclude that the present population structure is mainly a consequence of the species philopatric behaviour over a long time period instead of recent population bottlenecks. Based on our results, the Finnish population seems to have ongoing immigration from neighbouring populations. Hence, even though the population has recovered mainly through local growth, our results suggest that gene flow from genetically differentiated populations have had an impact as well.     

Are shed hair genomes the most effective noninvasive resource for estimating relationships in the wild?

1. Knowledge of relationships in wild populations is critical for better understanding mating systems and inbreeding scenarios to inform conservation strategies for endangered species. To delineate pedigrees in wild populations, study genetic connectivity, study genotype‐phenotype associations, trace individuals, or track wildlife trade, many identified individuals need to be genotyped at thousands of loci, mostly from noninvasive samples. This requires us to (a) identify the most common noninvasive sample available from identified individuals, (b) assess the ability to acquire genome‐wide data from such samples, and (c) evaluate the quality of such genome‐wide data, and its ability to reconstruct relationships between animals within a population.
2. We followed identified individuals from a wild endangered tiger population and found that shed hair samples were the most common compared to scat samples, opportunistically found carcasses, and opportunistic invasive samples. We extracted DNA from these samples, prepared whole genome sequencing libraries, and sequenced genomes from these.
3. Whole genome sequencing methods resulted in between 25%–98% of the genome sequenced for five such samples. Exploratory population genetic analyses revealed that these data were free of holistic biases and could recover expected population structure and relatedness. Mitochondrial genomes recovered matrilineages in accordance with long‐term monitoring data. Even with just five samples, we were able to uncover the matrilineage for three individuals with unknown ancestry.
4. In summary, we demonstrated that noninvasive shed hair samples yield adequate quality and quantity of DNA in conjunction with sensitive library preparation methods, and provide reliable data from hundreds of thousands of SNPs across the genome. This makes shed hair an ideal noninvasive resource for studying individual‐based genetics of elusive endangered species in the wild.

     

High genetic diversity and no inbreeding in the endangered copper redhorse, Moxostoma hubbsi (Catostomidae, Pisces): the positive sides of a long generation time

The evolutionary potential of a species is determined by its genetic diversity. Thus, management plans should integrate genetic concerns into active conservation efforts. The copper redhorse (Moxostoma hubbsi) is an endangered species, with an endemic distribution limited to the Richelieu River and a short section of the St Lawrence River in Quebec, Canada. The population, gradually fragmented since 1849, is characterized by a decline in population size and a lack of recruitment. A total of 269 samples were collected between 1984 and 2004 and genotyped using 22 microsatellite loci, which indicated that these fish comprise a single population, with a global F(ST) value of only 0.0038. Despite a small census size ( approximately 500), a high degree of genetic diversity was observed compared to common values for freshwater fishes (average number of 12.5 alleles/locus and average HO of 0.77 +/- 0.08). No difference was observed between expected and observed pairwise values of relatedness (r(xy): -0.00013 +/- 0.11737), suggesting an outbred population. Long-term Ne was estimated at 4476 whereas contemporary Ne values ranged from 107 to 568, suggesting a pronounced yet gradual demographic decline of the population, as no bottleneck could be detected for the recent past. By means of simulations, we estimated Ne would need to remain at more than approximately 400 to retain 90% of the genetic diversity over 100 years. Overall, these observations corroborate other recent empirical studies confirming that long generation times may act as a buffering effect contributing to a reduction in the pace of genetic diversity erosion in threatened species.     

Conservation genetics of a peripherally isolated population of the wood turtle (<Emphasis Type="Italic">Glyptemys insculpta</Emphasis>) in Iowa

The North American wood turtle, Glyptemys insculpta, is a semi-aquatic species that is considered rare, threatened, or endangered over much of its range. In this study, a particularly vulnerable peripheral isolate population in Iowa has been monitored over a period of 7 years. Population census size, estimated from mark-recapture data, and age structure determined from morphology are compared with genetic variation assessed using microsatellites. For reference, the genetics and demographics of this peripheral isolate are compared to data from a more dense population nearer the core of the species range in West Virginia. Geneflow between the Iowa population and a nearby population in Minnesota also is assessed. Genetic data indicate that the Iowa population is isolated, unique, and diverse. Although the Iowa population has lower allelic richness, lower heterozygosity, and smaller genetic effective population size than does the West Virginia population, the difference is not dramatic despite its lower population size, position at the periphery of the species range, and biogeographic history. The Iowa population is not inbred, and there is no genetic signature of a recent population bottleneck. However, interpretations of recent population dynamics based on genetic data may be unduly encouraging in long-lived species such as G. insculpta. Field data suggest a nearly complete lack of recruitment in Iowa. A number of environmental and anthropogenic factors, including recent increases in summer flooding during egg incubation, may have a more negative impact on the Iowa population than on the West Virginia population.     

Genetic composition of the Jadro softmouth trout following translocation into a new habitat

Genetic founder effects of a historical translocation (1965; app. 12 generations ago) of endangered softmouth trout from the only remaining indigenous population to an adjacent uninhabited river were investigated. A comparison of 15 microsatellites from contemporary samples in both the source and re-established populations indicated a very low level of genetic diversity within the two populations. Furthermore, considerable differences in estimates of variability and effective population sizes were detected between populations, but no recent bottlenecks were evident. Our data suggest that the re-established population has lost variation due to genetic drift associated with founder effects following the translocation. Accordingly, in relation to management, we suggest that the re-established population could serve as a secondary source of individuals to buffer possible extinction due to demographic events. Finally, conservation initiatives to enhance the census population size and increase levels of variability in the re-established population are proposed. Electronic Supplementary Material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Genetic status of the wood stork (Mycteria americana) from the southeastern United States and the Brazilian Pantanal as revealed by mitochondrial DNA analysis

The wood stork (Mycteria americana) is a colonial wading bird that inhabits the Neotropical region from the southeastern United States (US) to northern Argentina. The species is considered to be endangered in the US due to degradation of its foraging and breeding habitat. In other parts of its range, such as in the Brazilian Pantanal region, breeding populations of this species appear to be stable. We compared the levels of genetic variability and population structuring of the US and the Pantanal breeding populations using mitochondrial DNA (mtDNA) control region sequences. Twenty-seven haplotypes were identified among 88 wood stork samples collected from eight breeding colonies in the US and eight in the Pantanal. Patterns indicative of heteroplasmy were observed in 35.3% of the mtDNA sequences that were examined. Significantly higher levels of haplotype diversity were observed in the Pantanal samples compared to those from the US, suggesting that during the last century, demographic declines or a recent evolutionary bottleneck reduced the levels of mtDNA variability of the US population. Analyses of genetic structuring revealed non-significant genetic differentiation between these regions, indicating that either the populations were only recently separated or that gene flow continues to occur at low levels. Haplotype network analysis indicated low current levels of gene flow between populations that were closely related in the past.     

Monitoring population productivity in the saiga antelope

Effective conservation requires a good understanding of factors causing variation in population growth rate. We here analyse the relationship between female age and fecundity in the saiga antelope Saiga tatarica tatarica , a critically endangered ungulate of the Eurasian steppes and semideserts, at both individual and population levels. Annual variation in age structure and twinning rates was investigated using long-term datasets, sampling a total of 3308 females in four populations over more than 40 years. Further, a new non-invasive method is presented, estimating twinning rates from both calves and placentas encountered during calving aggregation transects. At an individual level, the most parsimonious model for twinning rates included three age classes (1, 2 and ≥3 years); however, the model with only two classes (1 and ≥2 years) was competitive and particularly useful for monitoring because these two age classes can reliably be determined by direct observation in the field. Among yearlings, 77.4% were fecund and 11.7% twinned, whereas among older females 94.6% were fecund and 72.6% twinned. At a population level, annual variation in age structure (proportion ≥2 years) correlated well with annual variation in twinning rate except in the north-west Pre-Caspian population. Our results suggest that the recent poaching-driven collapse in saiga numbers has potentially resulted in reductions in fecundity, which will have an impact on population growth rate. Our results highlight the potential for monitoring of twinning rate using non-invasive calving aggregation transects as a cost-effective additional tool to population counts for monitoring the status of this critically endangered species. These monitoring methods are also potentially transferable to other ungulate species.     

Signature of a Pre-Human Population Decline in the Critically Endangered Reunion Island Endemic Forest Bird Coracina newtoni

The exceptional biodiversity of Reunion Island is threatened by anthropogenic landscape changes that took place during the 350 years of human colonization. During this period the human population size increased dramatically from 250 to 800,000. The arrival of humans together with the development of agriculture, invasive species such as rats and cats, and deforestation has lead to the extinction of more than half of the original vertebrate species of the island. For the remaining species, significant work is being carried out to identify threats and conservation status, but little genetic work has been carried on some of the most endangered species. In the last decade theoretical studies have shown the ability of neutral genetic markers to infer the demographic history of endangered species and identify and date past population size changes (expansions or bottlenecks). In this study we provide the first genetic data on the critically endangered species the Reunion cuckoo-shrike Coracina newtoni. The Reunion cuckoo-shrike is a rare endemic forest bird surviving in a restricted 12-km(2) area of forested uplands and mountains. The total known population consists of less than one hundred individuals out of which 45 were genotyped using seventeen polymorphic microsatellite loci. We found a limited level of genetic variability and weak population structure, probably due to the limited geographic distribution. Using Bayesian methods, we identified a strong decline in population size during the Holocene, most likely caused by an ancient climatic or volcanic event around 5000 years ago. This result was surprising as it appeared in apparent contradiction with the accepted theory of recent population collapse due to deforestation and predator introduction. These results suggest that new methods allowing for more complex demographic models are necessary to reconstruct the demographic history of populations.     

西双版纳不同斑块望天树种群的密度、结构和生物量

 根据5个不同斑块的野外调查数据,分析了残存分布在西双版纳的国家一级保护稀有树种望天树（Parashorea chinensis）的种群密度与数量、 年龄结构与生物量动态, 组建了不同生长发育阶段的望天树个体生长与年龄的回归模型、个体生物量模型及种群年龄结构模型, 编制了不同 斑块的望天树种群及整个种群的静态生命表、存活曲线和年龄结构图。结果表明,不同斑块的望天树种群因种群年龄及所受到的干扰方式的不 同,其种群密度和年龄结构差异很大,不同年龄阶段的死亡率也不同。面积最小的斑块缺乏成熟个体,并出现龄级结构缺省的现象。不同斑块 局部种群的生物量随林龄的变化近似于Logistic增长,但各斑块局部种群的最大生物量以及生物量随时间的动态变化有所不同。整个种群的年 龄结构为稳定增长型种群,1～60龄的种群个体的死亡率随林龄的增加而下降,60～150龄的个体死亡率随林龄的增加而上升,180 龄后种群呈 现生理衰退,个体出现死亡高峰。种群的生物量在180 龄前呈Logistic 增长,此后,生物量下降。部分斑块受到严重的人为干扰,已严重威胁 其局部种群的生存。     

Reduced Genetic Diversity and Effective Population Size in an Endangered Atlantic Salmon (Salmo Salar) Population from Maine, USA

Atlantic salmon (Salmo salar) populations in Maine, USA, are listed as a Distinct Population Segment under the U.S. Endangered Species Act due to reduced spawning runs and juvenile densities. Whenever possible, optimal conservation strategies for endangered populations should incorporate both present and historical knowledge of genetic variation. We assayed genetic diversity at seven microsatellite loci and at the mitochondrial ND1 gene in an endangered wild population of Atlantic salmon captured from the Dennys River from 1963 to 2001 using DNA’s extracted from archival scale and tissue samples. We examined temporal trends of genetic diversity, population structure, and effective population size (Ne). Overall temporal trends of diversity and Ne show significant reductions from 1963 to 2001 raising the possibility that current restoration efforts may be impacted by historical loss of diversity potentially critical to adaptation. Although our results suggest genetic stability in this population from 1963 to 1981, significant differentiation was observed for both the 1995 and 2001 samples compared with all other temporal samples. The presence of an ND1 mtDNA haplotype in this population, historically observed only in European and Newfoundland stocks, may represent previously unrecognized local wild diversity or, alternatively, may represent introgression from non-native fish.     

珍稀濒危植物珙桐的生存与人为活动的关系

近年来,由于人类活动的加剧,区域开发项目的增多,致使珍稀濒危树种珙桐的天然分布面积和天然种群数量急剧下降,现天然分布种群已濒临灭绝。为此,本文系统分析了珙桐在不同地质阶段和不同历史阶段的地理分布状况,探讨了促使其地理分布及种数数理动态变化的主要原因,通过大面积的野外调查和查阅资料,提出了珙桐的保护措施。     

Origins of recently re‐established and newly discovered populations of the endangered butterfly Shijimiaeoides divinus (Lepidoptera: Lycaenidae) in Oita Prefecture,Japan

The endangered butterfly Shijimiaeoides divinus was believed to have been extirpated from Oita Prefecture, Kyushu, Japan, but was rediscovered in Taketa in recent years. This population is considered to have re‐established as a result of natural dispersal from Kumamoto, a neighboring prefecture located to the west of Oita. Furthermore, another population was recently found in Yufu, Oita Prefecture, which is an area where the species had never been recorded. To elucidate the origins of these two populations newly found from Oita Prefecture, their DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene were compared with those of other S. divinus populations from Kumamoto Prefecture, Honshu and Korea. The results supported the hypothesis that the Taketa population originated from Kumamoto Prefecture. However, it was not clear whether this population originated from the natural dispersal or deliberate release of individuals. It was also found that the Yufu population was not established by the deliberate release of individuals from Honshu or Korea; however, it remained unclear whether the population of S. divinus was native to Yufu, or originated from other localities in Kyushu.     

Bowhead whales in East Greenland,summers 2006–2008

During the summers of 2006–2008 extensive ship- and aircraft-based surveys were carried out in the Greenland Sea off Northeast Greenland. This resulted in 13 observations in a total of 17 bowhead whales Balaena mysticetus of the very small and critically endangered Spitsbergen stock. Although the observed number of whales seems high compared to previous surveys it is not possible to infer a population increase, due to the high observation effort in recent years. However, the observation of a young individual is encouraging as it seems to be the first observed young whale in this stock for decades.