Molecular Evidence for a Unique Evolutionary Lineage of Endangered Sonoran Desert Fish (Genus Poeciliopsis)

Efforts to restore an endangered species in its former range should be based on a sound understanding of evolutionary relationships among remaining natural populations. In this study mitochondrial (mt) DNA diversity within and among Gila River drainage populations of the endangered Sonoran topminnow ( Poeciliopsis occidentalis ) in Arizona was compared to that from neighboring populations in Sonora, Mexico, where the species remains locally abundant. No mtDNA diversity was detected within or among samples from the Gila River basin in Arizona. But considerable variation was found within and among populations from several river systems in Sonora. Examination of mtDNA from a population that inhabits the upper reaches of the Río Yaqui in southeastern Arizona revealed substantial divergence between it and all other populations examined. We comment on the implications of this divergent population for topminnow management in Arizona and argue for more-detailed genetic and morphological studies to determine the distributional limits and specific status of this highly divergent form.     

Diagnosing Units of Conservation Management

Species-oriented conservation programs attempt to analyze and maintain intra-specific variation in order to maximally preserve biological diversity. The "evolutionarily significant unit" has become an operational term for a group of organisms that should be the minimial unit for conservation management. No generally accepted definition for this term exists that would be the basis for the evaluation of these units in practical conservation situations. Currently, taxonomic decisions in species conservation are mostly based on the biological species concept. But the universal application of criteria of reproductive isolation or phenetic similarity to delimit conservation units is problematical. We favor a definition for evolutionarily significant units based on patterns of variation. In the theoretical framework of the phylogenetic species concept, conservation units are delimited by characters that diagnose clusters of individuals or populations to the exclusion of other such clusters. Characters are used for cladistic analysis to infer hypotheses of the phylogenetic relationships of individuals, and differentiated populations are diagnosed using population aggregation analysis. Characters can be based on genetic, morphological, ecological, or behavioral information, provided they are inferred to be heritable. The use of cladistics and population aggregation analysis has the potential to make the evaluation of evoluntionarily significant units objective and testable, an important consideration in politically controversial cases. Our cladistic approach is demonstrated by the evaluation of potential conservation units in the endangered tiger beetles Cicindela dorsalis and C. puritana .     

Using survival analysis to study translocation success in the Gila topminnow (Poeciliopsis occidentalis).

Translocation, the intentional release of captive-propagated and/or wild-caught animals into the wild in an attempt to establish, reestablish, or augment a population, is a commonly used approach to species conservation. Despite the frequent mention of translocation as an aid in threatened or endangered species recovery plans, translocations have resulted in the establishment of few sustainable populations. To improve the effectiveness of translocation efforts, it is essential to identify and adopt features that contribute to successful translocations. This study analyzed 148 translocations of the endangered Gila topminnow (Poeciliopsis occidentalis) to identify various factors that have significantly influenced translocation success. We quantified success as the "persistence time" of translocated populations and used survival analysis to interpret the role of several factors. The following factors affected persistence times of translocated populations: season in which the fish were translocated, habitat type of the translocation site, and genetic origin of the fish stocked. In general, factors associated with stocking, the population stocked, and the site of translocation can significantly affect the persistence of translocated populations and thus increase the probability of translocation success. For Gila topminnow, future translocations should be undertaken in late summer or fall (not early summer), should occur into ponds (not streams, wells, or tanks), and should generally utilize individuals from genetic lineages other than Monkey Spring. For other species, a key lesson emerging from this work is that life history attributes for each translocated species need to be considered carefully.     

Fitness in the Endangered Gila Topminnow

The Gila topminnow (Poeciliopsis o. occidentalis) is a small, live-bearing, endangered fish extant in a maximum of nine locales in four separate watersheds in the United States. To determine if these populations differed in their fitness, we obtained samples from the four watersheds and examined them for four fitness correlates: survival, growth rate, fecundity, and bilateral asymmetry. Earlier research found that one population, Sharp Spring, had higher allozyme heterozygosity than the other three and had higher survival, growth rate, and fecundity and lower bilateral asymmetry than a sample from one of the other populations with no polymorphic allozyme loci, Monkey Spring. We also verified that Sharp Spring fish were polymorphic for the same allozyme loci whereas the Monkey Spring population was not. We did not, however, find positive associations of allozyme heterozygosity with the four fitness correlates for the four samples. Because the earlier study had much lower survival, it is likely that the differences resulted from differential response of the two populations to a stressful laboratory environment. Whether this unknown stress occurs in natural environments or its effect is predictive of other stresses remains unresolved. As a result, we concur with suggestions in the draft recovery plan that topminnows from nearby sources be used for reintroductions and that the Sharp Spring stock not be used outside the upper Santa Cruz River drainage.     

Microsatellite variation and significant population genetic structure of endangered finless porpoises (Neophocaena phocaenoides) in Chinese coastal waters and the Yangtze River

The finless porpoise (Neophocaena phocaenoides) inhabits a wide range of tropical and temperate waters of the Indo-Pacific region. Genetic structure of finless porpoises in Chinese waters in three regions (Yangtze River, Yellow Sea, and South China Sea) was analyzed, including the Yangtze finless porpoise which is widely known because of its highly endangered status and unusual adaptation to freshwater. To assist in conservation and management of this species, ten microsatellite loci were used to genotype 125 individuals from the three regions. Contrary to the low genetic diversity revealed in previous mtDNA control region sequence analyses, relatively high levels of genetic variation in microsatellite profiles (H _E = 0.732–0.795) were found. Bayesian clustering analysis suggested that finless porpoises in Chinese waters could be described as three distinct genetic groups, which corresponded well to population “units” (populations, subspecies, or species) delimited in earlier studies, based on morphological variation, distribution, and genetic analyses. Genetic differentiation between regions was significant, with F _ST values ranging from 0.07 to 0.137. Immigration rates estimated using a Bayesian method and population ancestry analyses suggested no or very limited gene flow among regional types, even in the area of overlap between types. These results strongly support the classification of porpoises in these regions into distinct evolutionarily significant units, including at least two separate species, and therefore they should be treated as different management units in the design and implementation of conservation programmes.     

The Importance of Systematic Biology in Defining Units of Conservation

Abstract: Conservation biology is linked inextricably with systematic biology. The principles of systematic biology, however, have not been integrated completely into the practice and principles of conservation biology. Systematists have recognized for some time that a number of evolutionary processes lead to the diversification of lineages. Yet some present units of conservation, such as the evolutionarily significant unit ( Waples 1991), primarily emphasize only one of these processes, adaptation. Allopatric speciation produces biodiversity without requiring any adaptive shift (and consequent adaptive differences between daughter species), so definitions of conservation units that emphasize adaptation may underestimate biodiversity. We estimated the frequency of different modes of speciation for three groups of vertebrates. The frequency of allopatric speciation varies among these groups, but is an important type of speciation in two of the three groups studied. Our results, and the results of other published studies of the frequency of modes of speciation, demonstrate that any unit of conservation defined solely in terms of adaptation is likely to underestimate biological diversity.     

A Freshwater Classification Approach for Biodiversity Conservation Planning

Abstract:  Freshwater biodiversity is highly endangered and faces increasing threats worldwide. To be complete, regional plans that identify critical areas for conservation must capture representative components of freshwater biodiversity as well as rare and endangered species. We present a spatially hierarchical approach to classify freshwater systems to create a coarse filter to capture representative freshwater biodiversity in regional conservation plans. The classification framework has four levels that we described using abiotic factors within a zoogeographic context and mapped in a geographic information system. Methods to classify and map units are flexible and can be automated where high-quality spatial data exist, or can be manually developed where such data are not available. Products include a spatially comprehensive inventory of mapped and classified units that can be used remotely to characterize regional patterns of aquatic ecosystems. We provide examples of classification procedures in data-rich and data-poor regions from the Columbia River Basin in the Pacific Northwest of North America and the upper Paraguay River in central South America. The approach, which has been applied in North, Central, and South America, provides a relatively rapid and pragmatic way to account for representative freshwater biodiversity at scales appropriate to regional assessments.     

Conservation Implications of Drastic Reductions in the Smallest and Most Isolated Populations of Giant Pandas

Abstract: In conservation biology, understanding the causes of endangerment is a key step to devising effective conservation strategies. We used molecular evidence (coalescent simulations of population changes from microsatellite data) and historical information (habitat and human population changes) to investigate how the most‐isolated populations of giant pandas (Ailuropoda melanoleuca) in the Xiaoxiangling Mountains became highly endangered. These populations experienced a strong, recent demographic reduction (60‐fold), starting approximately 250 years BP. Explosion of the human population and use of non‐native crop species at the peak of the Qing Empire resulted in land‐use changes, deforestation, and habitat fragmentation, which are likely to have led to the drastic reduction of the most‐isolated populations of giant pandas. We predict that demographic, genetic, and environmental factors will lead to extinction of giant pandas in the Xiaoxiangling Mountains in the future if the population remains isolated. Therefore, a targeted conservation action—translocation—has been proposed and is being implemented by the Chinese goverment.     

Phylogeography and Limited Genetic Connectivity in the Endangered Boring Giant Clam across the Coral Triangle

Abstract: The Coral Triangle is the global center of marine biodiversity; however, its coral reefs are critically threatened. Because of the bipartite life history of many marine species with sedentary adults and dispersive pelagic larvae, designing effective marine protected areas requires an understanding of patterns of larval dispersal and connectivity among geographically discrete populations. We used mtDNA sequence data to examine patterns of genetic connectivity in the boring giant clam (Tridacna crocea) in an effort to guide conservation efforts within the Coral Triangle. We collected an approximately 485 base pair fragment of mtDNA cytochrome c oxidase 1 (CO1) from 414 individuals at 26 sites across Indonesia. Genetic structure was strong between regions (φ_ST=0.549, p < 0.00001) with 3 strongly supported clades: one restricted to western Sumatra, another distributed across central Indonesia, and a third limited to eastern Indonesia and Papua. Even within the single largest clade, small but significant genetic structure was documented (φ_ST=0.069, p < 0.00001), which indicates limited gene flow within and among phylogeographic regions. Significant patterns of isolation by distance indicated an average dispersal distance of only 25–50 km, which is far below dispersal predictions of 406–708 km derived from estimates of passive dispersal over 10 days via surface currents. The strong regional genetic structure we found indicates potent limits to genetic and demographic connectivity for this species throughout the Coral Triangle and provides a regional context for conservation planning. The recovery of 3 distinct evolutionarily significant units within a well‐studied taxonomic group suggests that biodiversity in this region may be significantly underestimated and that Tridacna taxa may be more endangered than currently recognized.     

Quantitative and Molecular Genetic Variation in Captive Cotton-Top Tamarins (Saguinus oedipus)

Most genetic surveys of captive and endangered populations are carried out with single gene characters bearing no direct relationship to life history or other features for which genetic variation needs to be maintained. Quantitative genetic estimates of heritable variation for life-history traits may be a more direct and appropriate measure of genetic variation for some conservation purposes. Furthermore, recent theoretical and empirical results indicate that genetic variation measured on these two levels may not be concordant. We analyzed heterozygosity at 41 allozyme loci and heritability for body weight in captive cotton-top tamarins ( Saguinus oedipus ) from the Marmoset Research Center of the Oak Ridge Associated Universities in order to compare these two levels of genetic variation. Cotton-top tamarins are a highly endangered species native to Colombia. Many animals currently reside in research facilities and zoological parks. A total of 106 animals were used in the isozyme survey, while data on 364 animals contributed to the quantitative genetic study of body weight. We found a very low average heterozygosity ( H = 1%) for this colony. Body weight was moderately and significantly heritable ( h ² = 35%). This heritability is within the normal range for natural animal populations. The finding of biologically significant levels of heritability in a population with abnormally low allozyme heterozygosity illustrates the point that low levels of allozyme heterozygosity should not be taken as an indication of overall lack of genetic variation in important quantitative characters such as life-history traits. Genetic variation required for adaptation of species to future environmental challenges can exist despite low levels of enzyme heterozygosity.     

Emerging prion disease drives host selection in a wildlife population

Infectious diseases are increasingly recognized as an important force driving population dynamics, conservation biology, and natural selection in wildlife populations. Infectious agents have been implicated in the decline of small or endangered populations and may act to constrain population size, distribution, growth rates, or migration patterns. Further, diseases may provide selective pressures that shape the genetic diversity of populations or species. Thus, understanding disease dynamics and selective pressures from pathogens is crucial to understanding population processes, managing wildlife diseases, and conserving biological diversity. There is ample evidence that variation in the prion protein gene (PRNP) impacts host susceptibility to prion diseases. Still, little is known about how genetic differences might influence natural selection within wildlife populations. Here we link genetic variation with differential susceptibility of white-tailed deer to chronic wasting disease (CWD), with implications for fitness and disease-driven genetic selection. We developed a single nucleotide polymorphism (SNP) assay to efficiently genotype deer at the locus of interest (in the 96th codon of the PRNP gene). Then, using a Bayesian modeling approach, we found that the more susceptible genotype had over four times greater risk of CWD infection; and, once infected, deer with the resistant genotype survived 49% longer (8.25 more months). We used these epidemiological parameters in a multi-stage population matrix model to evaluate relative fitness based on genotype-specific population growth rates. The differences in disease infection and mortality rates allowed genetically resistant deer to achieve higher population growth and obtain a long-term fitness advantage, which translated into a selection coefficient of over 1% favoring the CWD-resistant genotype. This selective pressure suggests that the resistant allele could become dominant in the population within an evolutionarily short time frame. Our work provides a rare example of a quantifiable disease-driven selection process in a wildlife population, demonstrating the potential for infectious diseases to alter host populations. This will have direct bearing on the epidemiology, dynamics, and future trends in CWD transmission and spread. Understanding genotype-specific epidemiology will improve predictive models and inform management strategies for CWD-affected cervid populations.     

Post-Perturbation Genetic Changes in Populations of Endangered Virgin River Chubs

A 34-kilometer reach of the Virgin River, Utah-Arizona-Nevada, was poisoned with rotenone in an attempt to eradicate non-native red shiners ( Cyprinella lutrensis ), a species implicated in the decline of native fish populations in the American West. An error in detoxification resulted in lethal concentrations of piscicide passing through an additional 50 kilometers of stream. We used allozyme electrophoresis to analyze genetic variation among pre- and post-poison samples of endangered Virgin River chubs ( Gila seminuda ). Pre-poison samples indicated a single panmictic population in the river. In contrast, fish subsequently produced through natural recruitment in poisoned reaches exhibited deviations from the original pattern of genetic variation. A genetic bottleneck caused by severe reduction in the number of spawning adults was indicated. The altered pattern persisted 2.5 years post-poisoning, indicating unexpectedly slow recolonization from the unpoisoned reach upstream. Genetic variation among hatchery-produced young was similarly unrepresentative of the original pattern because of the small number of brood fish used in propagation. Because of their small numbers and/or restricted distribution, endangered species are particularly vulnerable to natural or anthropogenic catastrophes. Assessment of the genetic impact of such events is essential but requires that baseline data are available.     

Evolutionary Distinctiveness,Threat Status,and Ecological Oddity in Primates

Abstract: The EDGE (evolutionarily distinct and globally endangered) conservation program ( http://www.edgeofexistence.org ) uses a composite measure of threat and phylogenetic isolation to rank species for conservation attention. Using primates as a test case, we examined how species that rank highly with this metric represent the collective from which they are drawn. We considered the ecological and morphological traits, including body mass, diet, terrestriality, and home range size, of all 233 species of primates. Overall, EDGE score and the level of deviance from the mean of 20 different ecological, reproductive, and morphological variables were correlated (mean correlation r =0.14, combined p =1.7 × 10^?14). Although primates with a high EDGE score had characteristics that made them seem odd, they did not seem to express more ancestral characteristics than expected. Sets of primate species with high EDGE scores will, therefore, collectively capture a broader than expected range of the biology of the clade. If similar patterns hold in other groups, the EDGE metric may be useful for prioritizing biodiversity for conservation.     

Critique of the Evolutionarily Significant Unit as a Definition for "Distinct Population Segments" under the U.S. Endangered Species Act

The U.S. Endangered Species Act grants protection to species, subspecies, and "distinct population segments" of vertebrate species. Historically, Congress included distinct population segments into endangered species legislation to enable the U.S. Fish and Wildlife Service to implement a flexible and pragmatic approach in listing populations of vertebrate species. Recently, the U.S. Fish and Wildlife Service and the National Marine Fisheries Service have proposed a policy that would narrowly define distinct population segments as evolutionarily significant units based on morphological and genetic distinctiveness between populations. Historically, the power to list species or populations as distinct population segments has been used to tailor management practices to unique circumstances; grant varied levels of protection in different parts of a species' range; protect species from extinction in significant portions of their ranges as well as to protect populations that are unique evolutionary entities. A strict redefinition of distinct population segments as evolutionarily significant units will compromise management efforts because the role of demographic and behavioral data will be reduced. Furthermore, strictly cultural, economic, or geographic justifications for listing populations as threatened or endangered will be greatly curtailed.     

Impact of Hybridization on a Threatened Trout of the Southwestern United States

Trouts native to the American Southwest provide an excellent example of the plight of endangered fishes from this region. The native species, Apache trout and Gila trout ( Oncorhynchus apache and O. gilae , respectively) have faced drastic reduction in habitat and detrimental interactions with introduced species, resulting in a dramatic decrease in numbers and sizes of populations. We used biochemical methods to identify diagnostic markers for the estimation of genetic relatedness and analysis of hybridization among native trouts and introduced cutthroat and rainbow trouts ( O. clarki and O. mykiss , respectively). Restriction endonuclease analysis of mitochondrial DNA (mtDNA) indicated that Apache and Gila trout were very similar to each other, and more similar to rainbow trout than cutthroat. Diagnostic allozyme marker loci indicated that Apache trout hybridized extensively with rainbows in four populations and provided no evidence for reproductive isolation between the forms. Analysis of mtDNA, however, indicated that introduced haplotypes were rare in these same individuals, identifying a bias in the direction of gene exchange between species. The potential reproductive isolation and lack of information concerning population structure necessitate further study of Apache trout to determine the appropriate management strategy for this threatened species. This case demonstrates that extreme care must be exercised when considering elimination of any contaminated population lest the unique genetic identity of the native taxon be lost forever.     

Molecular Systematics and the Conservation of Rare Species

Abstract: Despite the tradition of systematic biology as the science of diversity, systematics has until recently contributed relatively little to the theory and practice of conservation biology. We identify four areas in which systematics could contribute to the conservation of rare plant species: (1) species concepts, (2) the identification of lineages worthy of conservation, (3) the setting of conservation priorities, and (4) the effects of hybridization on the biology and conservation of rare species. Species concepts that incorporate history and reflect phylogeny ultimately will be more useful for preserving biodiversity than those that do not. Phylogenetic analyses involving conspecific populations often reveal multiple lineages that may warrant protection as evolutionarily distinct units. Phylogenetic information also should be considered in setting priorities for conservation. Systematics provides the tools for inferring relationships among organisms and, in conjunction with biogeography, for identifying those areas that harbor many actively speciating groups. Hybridization may lead to the extinction of a rare species, but in other cases, ironically, artificial hybridization with a more widespread congener may be the only way to preserve the gene pool of a rare species. We appeal to systematists to contribute actively to both conservation theory and practice, and we call for the integration of systematics in the establishment of conservation priorities and the development of strategies to preserve Earth's biota.     

Translating effects of inbreeding depression on component vital rates to overall population growth in endangered bighorn sheep

Evidence of inbreeding depression is commonly detected from the fitness traits of animals, yet its effects on population growth rates of endangered species are rarely assessed. We examined whether inbreeding depression was affecting Sierra Nevada bighorn sheep (Ovis canadensis sierrae), a subspecies listed as endangered under the U.S. Endangered Species Act. Our objectives were to characterize genetic variation in this subspecies; test whether inbreeding depression affects bighorn sheep vital rates (adult survival and female fecundity); evaluate whether inbreeding depression may limit subspecies recovery; and examine the potential for genetic management to increase population growth rates. Genetic variation in 4 populations of Sierra Nevada bighorn sheep was among the lowest reported for any wild bighorn sheep population, and our results suggest that inbreeding depression has reduced adult female fecundity. Despite this population sizes and growth rates predicted from matrix-based projection models demonstrated that inbreeding depression would not substantially inhibit the recovery of Sierra Nevada bighorn sheep populations in the next approximately 8 bighorn sheep generations (48 years). Furthermore, simulations of genetic rescue within the subspecies did not suggest that such activities would appreciably increase population sizes or growth rates during the period we modeled (10 bighorn sheep generations, 60 years). Only simulations that augmented the Mono Basin population with genetic variation from other subspecies, which is not currently a management option, predicted significant increases in population size. Although we recommend that recovery activities should minimize future losses of genetic variation, genetic effects within these endangered populations-either negative (inbreeding depression) or positive (within subspecies genetic rescue)-appear unlikely to dramatically compromise or stimulate short-term conservation efforts. The distinction between detecting the effects of inbreeding depression on a component vital rate (e.g., fecundity) and the effects of inbreeding depression on population growth underscores the importance of quantifying inbreeding costs relative to population dynamics to effectively manage endangered populations.     

Patterns of Genetic Diversity in Remaining Giant Panda Populations

Abstract: The giant panda ( Ailuropoda melanoleuca ) is among the more familiar symbols of species conservation. The protection of giant panda populations has been aided recently by the establishment of more and better-managed reserves in existing panda habitat located in six mountain ranges in western China. These remaining populations are becoming increasingly isolated from one another, however, leading to the concern that historic patterns of gene flow will be disrupted and that reduced population sizes will lead to diminished genetic variability. We analyzed four categories of molecular genetic markers (mtDNA restriction-fragment-length polymorphisms [RFLP], mtDNA control region sequences, nuclear multilocus DNA fingerprints, and microsatellite size variation) in giant pandas from three mountain populations (Qionglai, Minshan, and Qinling) to assess current levels of genetic diversity and to detect evidence of historic population subdivisions. The three populations had moderate levels of genetic diversity compared with similarly studied carnivores for all four gene measures, with a slight but consistent reduction in variability apparent in the smaller Qinling population. That population also showed significant differentiation consistent with its isolation since historic times. From a strictly genetic perspective, the giant panda species and the three populations look promising insofar as they have retained a large amount of genetic diversity in each population, although evidence of recent population reduction—likely from habitat loss—is apparent. Ecological management to increase habitat, population expansion, and gene flow would seem an effective strategy to stabilize the decline of this endangered species.     

Endangered Cacti in the Chihuahuan Desert: II. Biogeography and Conservation

The distributions of the majority of the endangered cacti in the Chihuahuan Desert Region are concentrated in the southeastern and eastern segments of the area, where the predominance of narrow endemism is a remarkable phytogeographic phenomenon. We used three criteria—species richness, conservation value, and complementarity—to evaluate 37 cactus-rich area units in the Chihuahuan Desert. The evaluation of these three quantitative parameters together allowed us to determine that seven of these areas (Huizache, Tolimán, Ciudad Victoria, Metztitlán, Cuatro Ciénegas, Jaumave, and Xichú) should be considered conservation priorities because they hold the most significant assemblage of endangered species, in terms of their numbers and their rarity. Conservation actions in these seven areas would protect 52 (55.9%) of the 93 endangered species studied here, most of which have extremely narrow distributions. Geographically, these critical areas are located in the Queretaroan-Hidalgoan arid zone (in the States of Querétaro, Hidalgo, and Guanajuato), in the Jaumave Valley (Tamaulipas), and in two disjunct areas (San Luis Potosí and Coahuila) in the interior of the Chihuahuan Desert.     

Human Impacts on Drainages of the Mesa Central, Mexico, and Its Genetic Effects on an Endangered Fish, Zoogoneticus quitzeoensis

Abstract:  The Mesa Central of Mexico is of special conservation interest due to its high richness of freshwater fish species, of which the goodeines are one of the most representative groups. Through an integrated approach, we determined conservation priorities for goodeine populations. We based our recommendations on the genetic diversity (variation in five microsatellite DNA loci) in 10 populations of Zoogoneticus quitzeoensis and on an analysis of ecological (e.g., presence of exotic species), social (e.g., political situation), and environmental (e.g., pollution) information for 52 historical occurrence points for species in the genus Zoogoneticus . Patterns of genetic erosion and genetic diversity indices were closely associated with human impact. Recent bottleneck events were most evident in the populations from remnants of the lakes drained at the beginning of the twentieth century. We identified seven operational conservation units (OCUs), all of which should be conserved because they contain unique portions of the total variation of the species. Special attention needs to be given to increase genetic variability, recover population sizes, and reestablish contact among populations within OCUs. It is imperative to create an integrative and effective approach for the recovery and conservation of the freshwater fish diversity of Central Mexico that is based on social and natural sciences.