Larval survival in populations of the large copper butterfly Lycaena dispar batavus

Lycaena dispar is recorded as an early case of extinction and subsequent re-introduction in the UK. However, repeated establishment attempts have not resulted in self-sustaining populations, including those having taken place at Woodwalton Fen (Cambridgeshire, UK).
Earlier studies at Woodwalton Fen showed that the highest mortalities occurred between the egg stage and resumption of larval feeding in spring.
This study was designed; firstly, to investigate factors causing field mortality during different larval stages; secondly, to compare survival on Rumex hydrolapathum foodplants in different habitat situations; and thirdly, to compare survival in a natural population (Weerribben, The Netherlands) with the introduced Woodwalton Fen population. Experiments employed exclusion cages in order to examine the relative roles of vertebrate and invertebrate natural enemies, and survival on food-plants in open fen and waterside situations was compared.
Results suggest invertebrate predation to be the dominant mortality factor acting upon pre-diapause larvae. Over the winter diapause natural enemies do not have a significant role. However during both these stages losses still occur that are unaccounted for. Extensive winter flooding appears to increase overwintering losses. Vertebrate predators cause significant mortality of post-diapause larvae.
No significant difference was found between larval survival on open fen versus waterside plants.
Comparison of introduced and native populations reveal that overwintering survival was significantly higher in the latter; potential reasons for this difference are discussed.     

Phylogeography, geographic structure, genetic variation, and potential species boundaries in Philippine slender toads

We investigated phylogeography of Philippine slender toads (genus Ansonia) and used a temporal framework for diversification, statistical tests of alternate topologies, and Bayesian approaches to test previous hypotheses concerning dispersal to, and colonization routes within, the southern Philippine island of Mindanao. Two species of Ansonia previously have been documented, with ranges separated by an east–west split corresponding to the approximate boundaries of Mindanao’s paleoisland precursors. We present new mtDNA sequence data (1946 bp from genes encoding ND1, 16S rRNA and tRNA^Leu) for 105 Ansonia specimens sampled from 20 localities on Mindanao Island. Our data suggest that Philippine Ansonia is composed of at least eight, well-supported population lineages, structured into a minimum of four highly divergent mtDNA clades. One clade corresponds to Ansonia mcgregori, a range-restricted species apparently limited to the distal portion of the Zamboanga Peninsula of western Mindanao. Two morphologically indistinguishable, but genetically divergent, lineages possibly are undescribed cryptic species from western Mindanao. We recognize the five remaining lineages as Ansonia muelleri pending data from morphology or bioacoustics that might diagnose separate species among these lineages. Regardless of their species status, the five allopatric lineages of A. muelleri should be viewed as important genetic units for future genetic conservation planning.     

Population genetic structure of the endangered crayfish Austropotamobius pallipes in France based on microsatellite variation: biogeographical inferences and conservation implications

1. One important goal in conservation biology is to characterise evolutionary lineages within endangered species before management decisions are taken. Here, we assess population differentiation in the freshwater crayfish Austropotamobius pallipes, an endangered species endemic to western Europe and provide valuable information for the conservation of French populations. 2. Analysis of five microsatellite loci in 44 populations revealed very different within population levels of genetic diversity (0.000 < H₀ < 0.564). Two groups, corresponding to northern and southern French populations, showed a high degree of genetic differentiation in both allele frequencies and allele sizes. Comparison of these results with previous studies of A. pallipes strongly suggests that the divergence between northern and southern populations could have occurred during the last glaciation period of the Pleistocene from one Atlantic and one Mediterranean refuge. 3. Evidence for genetic admixture between these two lineages was revealed by correspondence analyses in southern populations, probably as the result of artificial translocations. 4. French populations appeared significantly differentiated among the different river drainages and were highly structured within rivers. The impact of population size, population bottlenecks and founder events on the population genetic differentiation are discussed. 5. Based on these results, we propose the designation of two evolutionarily significant units for A. pallipes in France. Our data also support the maintenance of separate demographic management strategies for crayfish inhabiting different river systems. However, genetic analyses will have to be combined with demographic and ecological data for sustainable conservation programmes.     

Population genetic structure and conservation of marbled murrelets (<Emphasis Type="Italic">Brachyramphus marmoratus</Emphasis>)

Marbled murrelets (Brachyramphus marmoratus) are coastal seabirds that nest from California to the Aleutian Islands. They are declining and considered threatened in several regions. We compared variation in the mitochondrial control region, four nuclear introns and three microsatellite loci among194 murrelets from throughout their range except Washington and Oregon. Significant population genetic structure was found: nine private control region haplotypes and three private intron alleles occurred at high frequency in the Aleutians and California; global estimates of F _ST or Φ_ST and most pairwise estimates involving the Aleutians and/or California were significant; and marked isolation-by-distance was found. Given the available samples, murrelets appear to comprise five genetic management units: (1) western Aleutian Islands, (2) central Aleutian Islands, (3) mainland Alaska and British Columbia, (4) northern California, and (5) central California.     

The conservation of declining butterfly populations in Britain and Europe: priorities, problems and successes

The status, ecology and conservation of butterflies in Europe and Britain are reviewed, as a background to the National Trust's past and future contribution to British conservation. Britain has a poor butterfly fauna by European standards, the main areas of endemism and species richness being in the Alps and southern Europe. To date, the main declines among European butterfly populations have occurred across central-northern Europe, with slightly higher extinction rates in mainland countries than in Britain. The main causes of decline are biotope destruction, the loss of certain species' habitats within surviving semi-natural biotopes due to changed land management, and a failure by several species to track the patches of their habitat that are still being generated in modern fragmented landscapes. Until recently, most conservation programmes failed to take account of the latter two factors, resulting in many local extinctions of rare butterfly species even in conservation areas. Recent measures have been much more successful; many were first tested on National Trust properties.     

Phylogeography, genetic structure and population divergence time of cheetahs in Africa and Asia: evidence for long-term geographic isolates

The cheetah (Acinonyx jubatus) has been described as a species with low levels of genetic variation. This has been suggested to be the consequence of a demographic bottleneck 10 000–12 000 years ago (ya) and also led to the assumption that only small genetic differences exist between the described subspecies. However, analysing mitochondrial DNA and microsatellites in cheetah samples from most of the historic range of the species we found relatively deep phylogeographic breaks between some of the investigated populations, and most of the methods assessed divergence time estimates predating the postulated bottleneck. Mitochondrial DNA monophyly and overall levels of genetic differentiation support the distinctiveness of Northern‐East African cheetahs (Acinonyx jubatus soemmeringii). Moreover, combining archaeozoological and contemporary samples, we show that Asiatic cheetahs (Acinonyx jubatus venaticus) are unambiguously separated from African subspecies. Divergence time estimates from mitochondrial and nuclear data place the split between Asiatic and Southern African cheetahs (Acinonyx jubatus jubatus) at 32 000–67 000 ya using an average mammalian microsatellite mutation rate and at 4700–44 000 ya employing human microsatellite mutation rates. Cheetahs are vulnerable to extinction globally and critically endangered in their Asiatic range, where the last 70–110 individuals survive only in Iran. We demonstrate that these extant Iranian cheetahs are an autochthonous monophyletic population and the last representatives of the Asiatic subspecies A. j. venaticus. We advocate that conservation strategies should consider the uncovered independent evolutionary histories of Asiatic and African cheetahs, as well as among some African subspecies. This would facilitate the dual conservation priorities of maintaining locally adapted ecotypes and genetic diversity.     

Population structure and genetic diversity of the threatened quillwort Isoëtes malinverniana and implication for conservation

The goal of this research was to investigate genetic variation in Isoëtes malinverniana (Isoëtaceae) to select candidate populations for future conservation efforts. To this aim, ISSR and AFLP analyses, carried out using six and four primer combinations, respectively, produced a total of 425 bands, 97.18% of which were polymorphic.Our results suggest that I. malinverniana shows medium to high genetic diversity (mean Nei's genetic diversity: H = 0.1491 for ISSR data; H = 0.2289 for AFLP data) and a substantial amount of gene flow between the analysed populations (N_m = 1.768, with combined ISSR and AFLP data). The moderate levels of population differentiation support the hypothesis that the fragmentation and isolation of I. malinverniana occurred only recently, probably due to the intensive agriculture practice and water pollution.These results will be used to focus further studies aimed at supporting reintroduction programs within suitable sites of the distribution area.     

Assessment of genetic diversity in the critically endangered Australian corroboree frogs, Pseudophryne corroboree and Pseudophryne pengilleyi, identifies four evolutionarily significant units for conservation

The iconic and brightly coloured Australian northern corroboree frog, Pseudophryne pengilleyi , and the southern corroboree frog, Pseudophryne corroboree are critically endangered and may be extinct in the wild within 3 years. We have assembled samples that cover the current range of both species and applied hypervariable microsatellite markers and mitochondrial DNA sequences to assess the levels and patterns of genetic variation. The four loci used in the study were highly variable, the total number of alleles observed ranged from 13 to 30 and the average number of alleles per locus was 19. Expected heterozygosity of the four microsatellite loci across all populations was high and varied between 0.830 and 0.935. Bayesian clustering analyses in structure strongly supported four genetically distinct populations, which correspond exactly to the four main allopatric geographical regions in which the frogs are currently found. Individual analyses performed on the separate regions showed that breeding sites within these four regions could not be separated into distinct populations. Twelve mtND2 haplotypes were identified from 66 individuals from throughout the four geographical regions. A statistical parsimony network of mtDNA haplotypes shows two distinct groups, which correspond to the two species of corroboree frog, but with most of the haplotype diversity distributed in P. pengilleyi . These results demonstrate an unexpectedly high level of genetic diversity in both species. Our data have important implications for how the genetic diversity is managed in the future. The four evolutionarily significant units must be protected and maintained in captive breeding programmes for as long as it is possible to do.     

Bromeliaceae diversity and conservation in Minas Gerais state,Brazil

Field work and data from herbaria collections (2686 records) representing 283 taxa (265 species and 18 infraspecific taxa) of Bromeliaceae occurring at Minas Gerais state, southeastern Brazil, were analyzed in order to obtain distribution and diversity information, and to determine IUCN (The World Conservation Union) conservation status for each taxon. A map containing 1° × 1° grid cells was used to identify priority areas for new research collections, areas of high species diversity, and Bromeliaceae conservation status. A clear decrease in Bromeliaceae diversity is observed between the eastern and the western portions of Minas Gerais, and low floristic similarities were found between neighboring grid cells. The rocky mountains of Cadeia do Espinhaço are considered the most important area for Bromeliaceae endemics. From the 283 taxa of Bromeliaceae that occur at Minas Gerais, 118 (42%) are considered threatened, and 124 taxa (44% of the total) do not occur inside any protected area. The region of the Quadrilátero Ferrífero in the southern portion of the Cadeia do Espinhaço is the most threatened, and urgent strategies for conservation of this rich Bromeliaceae flora are needed. Northeastern Minas Gerais, particularly the rocky outcrops or inselbergs located in the Jequitinhonha and Mucuri rivers drainage basins need additional collection efforts and conservation actions focused on these saxicolous taxa.     

Determination of conservation priorities in regions with multiple political jurisdictions

Red lists serve as the most prominent tool for priority setting in applied conservation, even though they were not originally designed for this task. Hence, threat status does not always reflect actual conservation needs and can be very different from actual conservation priorities. Therefore, red lists may at best be a suboptimal tool for setting conservation priorities in a country or region. As a response, a range of alternative or complementary tools have been developed, with approaches, methods, and parameters such as population decline, population center etc. used, differing widely among countries. One recent development is the combination of conservation status with a measure of the international importance of a population in a focal region for the global survival of a species. Here, we provide a new method that integrates the two concepts while keeping them conceptually separate. The main benefit of this method is that it can be applied across variable geographical scales such as regions, countries, and even continents. Furthermore, it allows for better recommendations for applied conservation and conservation policy development than the two concepts in isolation. Our method, if applied internationally, would allow for a standardized priority setting in species conservation, would be highly comparable between countries, and would lead to a more efficient use of the limited financial and human resources for monitoring and conservation of biodiversity.     

A survey of equid mitochondrial DNA: Implications for the evolution, genetic diversity and conservation of Equus

The evolution, taxonomy and conservation of the genus Equuswere investigated by examining the mitochondrial DNA sequences of thecontrol region and 12S rRNA gene. The phylogenetic analysis of thesesequences provides further evidence that the deepest node in thephylogeny of the extant species is a divergence between twolineages; one leading to the ancestor of modern horses (E.ferus, domestic and przewalskii) and the other to thezebra and ass ancestor, with the later speciation events of the zebrasand asses occurring either as one or more rapid radiations, or withextensive secondary contact after speciation. Examination of the geneticdiversity within species suggested that two of the E. hemionussubspecies (E. h. onager and E. h. kulan) onlyrecently diverged, and perhaps, are insufficiently different to beclassified as separate subspecies. The genetic divergence betweendomestic and wild forms of E. ferus (horse) and E.africanus (African ass) was no greater than expected within anequid species. In E. burchelli (plains zebra) there was anindication of mtDNA divergence between populations increasing withdistance. The implications of these results for equid conservation arediscussed and recommendations are made for conservation action.     

Assessing genetic diversity for conservation management: a case study of a threatened reptile

The consequences of inbreeding in small isolated populations are well documented, yet populations are often managed in isolation to avoid irreversibly mixing genetic lineages and to maintain the historic integrity of each population. Three remaining populations of Whitaker's skink ( Cyclodina whitakeri ) in New Zealand, remnants of a once wider distribution, illustrate the conflict between this genetic goal (separate management of populations) with the more tangible and immediate threats of small population size and inbreeding. Middle and Castle Islands harbour populations of C. whitakeri and have been separated from each other and from the mainland for ∼10 000 years. The single mainland population at Pukerua Bay is extremely small, declining and deemed a high priority for management. We sequenced a 550 bp region of mitochondrial DNA (mtDNA,ND2) and genotyped animals from all three populations at 13 microsatellite loci. The population of C. whitakeri at Pukerua Bay showed marked differences from the island populations at both mtDNA (unique, fixed haplotype) and microsatellite loci ( F _ST∼0.20), and private alleles were detected at a high frequency (24% of all alleles). However, we attribute this pattern to an historic genetic gradient coupled with rapid genetic drift. Further, animals in captivity show genetic signatures of both Pukerua Bay and island populations, despite the goal to maintain a pure Pukerua Bay stock. The mixed genetic stock in captivity provides an opportunity for the addition of skinks from Middle Island to evaluate the risks of further population hybridization, including the disruption of potential local adaptation, while mitigating the risks of inbreeding.     

Intra-population genetic diversity of two wheatgrass species along altitude gradients on the Qinghai-Tibetan Plateau: its implication for conservation and utilization

A fluorescence-based AFLP fingerprinting was applied to investigate genetic diversity in 22 natural populations of two wheatgrasses from Qinghai-Tibetan Plateau at different altitudes: the hexaploid Elymus nutans Griseb and the tetraploid E. burchan-buddae (Nevski) Tzvelev (Poaceae). Five selective primer combinations used in this study generated a total of 637 AFLP fragments across all the samples, with 612 fragments in E. nutans and 570 in E. burchan-buddae. About 45% of the scored fragments were <200 bp and about 13% of the fragments were >400 bp. Results showed that genetic diversity within populations of the two Elymus species increased gradually with the increase in altitudes from the lowest sampling sites (2800 m) and reached a plateau at the medium altitudes, and then started to decrease with the increase in altitudes. Regression analysis demonstrated a clear pattern between the expected heterozygosity (H _e) or Shannon index (I) and altitude variation, where the highest H _e values (0.3449 for E. nutans and 0.3167 E. burchan-buddae) and I values (0.5123 and 0.4759) were expected at the altitudes 3399 m and 3418 m across all sampling sites, respectively for E. nutans and E. burchan-buddae. In other words, higher genetic diversity was observed in populations occurring at the medium altitudes (3200–3600 m) than those at the low and high altitudes for the two Elymus species. Principal coordinate analysis (PCA) did not show clear association between genetic relationships of populations and their occurrences at a particular altitude. The above results suggest that efforts for conservation and utilization of two wheatgrasses species should focus more on populations occurring at the medium altitudes.     

Genetic diversity of Dinaric brown bears (Ursus arctos) in Croatia with implications for bear conservation in Europe

Brown bears have lost most of their range on the European continent. The remaining western populations are small, isolated and highly endangered. The Dinaric-Pindos brown bear population is the western-most stable population and the fourth largest in Europe. It has been recognized as a potential source for recolonization of populations whose survival is at risk. Indeed, several translocations of Dinaric bears to Italy, Austria and France have recently been made. Despite the importance of the Dinaric bear population, its genetic status remains poorly understood. Using tissue samples from 156 hunted or accidentally killed Dinaric bears in Croatia, this study analysed genetic diversity at 12 microsatellite loci, as well as population structure and past reductions in size. In addition, a subset of 59 samples was used to assess diversity of the mitochondrial DNA control region. The results indicate that Dinaric bears have high nuclear genetic diversity, as compared to other extant brown bear populations, despite genetic evidence of a bottleneck caused by past persecutions. However, haplotype diversity was low, probably as a result of male-biased dispersal and female philopatry. Not surprisingly, no evidence of population sub-structure was found using nuclear markers, as the bear habitat has remained continuous and the highway network has been built only recently. Management should focus on maintaining habitat connectivity and keeping the effective population size as large as possible. In addition, when removing individuals, care should be taken not to further deplete the population of rare haplotypes. A coordinated transboundary management of the entire Dinaric-Pindos brown bear population should be a priority for its long-term conservation.     

Comparative survey of within-river genetic structure in Atlantic salmon; relevance for management and conservation

In wild populations, defining the spatial scale at which management and conservation practices should focus remains challenging. In Atlantic salmon, compelling evidence suggests that genetic structure within rivers occurs, casting doubt on the underlying premise of the river-based management approach for this species. However, no comparisons of within-river genetic structure across different systems have been performed yet to assess the generality of this pattern. We compared the within-river genetic structure of four important salmon rivers in North America and evaluated the extent of genetic differentiation among their main tributaries. We found a hierarchical genetic structure at the river and tributary levels in most water systems, except in the Miramichi where panmixia could not be rejected. In the other cases, genetic differentiation between most tributaries was significant and could be as high as that found between rivers of the same geographical region. More importantly, the extent of genetic differentiation between tributaries varied greatly among water systems, from well differentiated (θ_ST = 0.035) to undifferentiated (θ_ST = −0.0003), underlying the difficulty in generalizing the ubiquity of within-river genetic structure in Atlantic salmon. Thus, this study underlines the importance of evaluating the genetic structure of Atlantic salmon in large water systems on a case by case basis in order to define the most appropriate spatial scale and focal unit for efficient management and conservation actions. The potential consequences of management at an inappropriate spatial scale are discussed.     

Extremely low effective population sizes, genetic structuring and reduced genetic diversity in a threatened bumblebee species, Bombus sylvarum (Hymenoptera: Apidae)

Habitat fragmentation may severely affect survival of social insect populations as the number of nests per population, not the number of individuals, represents population size, hence they may be particularly prone to loss of genetic diversity. Erosion of genetic diversity may be particularly significant among social Hymenoptera such as bumblebees (Bombus spp.), as this group may be susceptible to diploid male production, a suggested direct cost of inbreeding. Here, for the first time, we assess genetic diversity and population structuring of a threatened bumblebee species (Bombus sylvarum) which exists in highly fragmented habitat (rather than oceanic) islands. Effective population sizes, estimated from identified sisterhoods, were very low (range 21-72) suggesting that isolated populations will be vulnerable to loss of genetic variation through drift. Evidence of significant genetic structuring between populations (theta = 0.084) was found, but evidence of a bottleneck was detected in only one population. Comparison across highly fragmented UK populations and a continental population (where this species is more widespread) revealed significant differences in allelic richness attributable to a high degree of genetic diversity in the continental population. While not directly related to population size, this is perhaps explained by the high degree of isolation between UK populations relative to continental populations. We suggest that populations now existing on isolated habitat islands were probably linked by stepping-stone populations prior to recent habitat loss.     

Restoration of coral populations in light of genetic diversity estimates

Due to the importance of preserving the genetic integrity of populations, strategies to restore damaged coral reefs should attempt to retain the allelic diversity of the disturbed population; however, genetic diversity estimates are not available for most coral populations. To provide a generalized estimate of genetic diversity (in terms of allelic richness) of scleractinian coral populations, the literature was surveyed for studies describing the genetic structure of coral populations using microsatellites. The mean number of alleles per locus across 72 surveyed scleractinian coral populations was 8.27 (±0.75 SE). In addition, population genetic datasets from four species (Acropora palmata, Montastraea cavernosa, Montastraea faveolata and Pocillopora damicornis) were analyzed to assess the minimum number of donor colonies required to retain specific proportions of the genetic diversity of the population. Rarefaction analysis of the population genetic datasets indicated that using 10 donor colonies randomly sampled from the original population would retain >50% of the allelic diversity, while 35 colonies would retain >90% of the original diversity. In general, scleractinian coral populations are genetically diverse and restoration methods utilizing few clonal genotypes to re-populate a reef will diminish the genetic integrity of the population. Coral restoration strategies using 10–35 randomly selected local donor colonies will retain at least 50–90% of the genetic diversity of the original population. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Habitat use of the endangered butterfly Euphydryas maturna and forestry in Central Europe

The knowledge of ecological requirements of declining butterflies of European woodlands remains limited, which hinders conservation management of their localities. This also applies for continentally threatened scarce fritillary Euphydryas maturna . On the basis of the largest data set on its habitat use ever collected in Central Europe, we analyse habitat requirements of its populations in Austria (A), the Czech Republic (Cz) and Germany (D). All studied populations inhabit open-canopy sites within woodlands, but larval survival decreases under full sun and preferred sites are relatively humid and sheltered. Nests of pre-hibernation larvae occur at terminal branches of Fraxinus excelsior , 1.5–3 m above the ground. Pre-hibernation mortality reaches 70% (Cz, D). Another limiting factor is quality of woodland vegetation: post-hibernation larvae consume a wide range of herbs and shrubs, and adult distribution is linked to nectar availability. The butterfly thus depends on highly heterogeneous early successional stages of deciduous woods, historically maintained by coppicing (Cz, D) and forest pasture (A). Restoration of these traditional methods offers the only chance for survival of E. maturna in Central Europe, and the butterfly may become a flagship for other threatened organisms of open-canopy woodlands.     

Gene diversity analysis of mitochondrial DNA, microsatellites and allozymes in landlocked Atlantic salmon

This study investigates the patterns of genetic diversity detected in allozymes, mtDNA, and microsatellites, in order to assess their relative efficacy to differentiate sympatric landlocked salmon populations and to estimate changes in genetic diversity between wild and first-generation hatchery fish. Overall, the three genetic markers indicated a genetic differentiation between two sympatric populations of Lake Saint-Jean, Québec. MtDNA and microsatellites also showed significant differences between wild and first-generation hatchery fish originating from the same river. Allozyme analysis was the most limited approach due to the low genetic diversity detected and the necessity to kill specimens. Although low polymorphism was found in mtDNA, it was the most discriminant marker between wild populations. Microsatellite analysis appears to be a promising approach due to its high sensitivity in differentiating wild populations, in detecting changes in allele composition between wild and first-generation hatchery fish and its potential for increased resolution by augmenting the number of polymorphic loci. Given the benefits and disadvantages of the three methods, the combination of mtDNA and microsatellite analyses will best address our research objectives.     

Genetic diversity, phylogeny and conservation of the Javan rhinoceros (Rhinoceros sondaicus)

With a total population of less than 60 individuals limited to two locations, the Javan rhinoceros is perhaps the most endangered large mammal on earth. Although species specific information is crucial to its conservation, its precarious status, habitat inaccessibility, and behavioral adaptations pose major obstacles to its study. Here we report on the first genetic analysis of the two extant populations, in Ujung Kulon, Indonesia, and Cat Tien, Vietnam, and discuss their conservation. As its critically endangered status precluded invasive sampling, we extracted DNA from dung, amplifying and sequencing segments of the mtDNA 12S rRNA gene and the non-coding D-loop. Divergence between Javan rhinos from Ujung Kulon and Cat Tien was similar to that between recognized subspecies of African rhinos, and exceeded that between Sumatran rhinos. The Ujung Kulon and Cat Tien populations represent separate Evolutionary Significant Units, advocating independent management. However, given the precariousness of the Cat Tien population, demographic considerations may override genetic issues in the short term. Genetic diversity of Javan rhinos was low and population expansion in the immediate future will be critical for its survival.