Patterns of nuclear and mitochondrial DNA variation in Iberian populations of <Emphasis Type="Italic">Emys orbicularis</Emphasis> (Emydidae): conservation implications

The European pond turtle (Emys orbicularis) is threatened and in decline in several regions of its natural range, due to habitat loss combined with population fragmentation. In this work, we have focused our efforts on studying the genetic diversity and structure of Iberian populations with a fine-scale sampling (254 turtles in 10 populations) and a representation from North Africa and Balearic island populations. Using both nuclear and mitochondrial markers (seven microsatellites, ∼1048 bp nDNA and ∼1500 bp mtDNA) we have carried out phylogenetic and demographic analyses. Our results show low values of genetic diversity at the mitochondrial level although our microsatellite dataset revealed relatively high levels of genetic variability with a latitudinal genetic trend decreasing from southern to northern populations. A moderate degree of genetic differentiation was estimated for Iberian populations (genetic distances, F _ST values and clusters in the Bayesian analysis). The results in this study combining mtDNA and nDNA, provide the most comprehensive population genetic data for E. orbicularis in the Iberian Peninsula. Our results suggest that Iberian populations within the Iberian–Moroccan lineage should be considered as a single subspecies with five management units, and emphasize the importance of habitat management rather than population reinforcement (i.e. captive breeding and reintroduction) in this long-lived species.     

Dietary shifts in two vultures after the demise of supplementary feeding stations: consequences of the EU sanitary legislation

Among vertebrates, specialization in scavenging has appeared only in “true” Gyps vultures, which usually base their diet almost exclusively on carcasses of medium and large-sized mammals, whereas all other scavengers rely on broader ranges of prey. The availability of food for scavengers in Western Europe has not been limited during recent decades permitting the existence and growth of huge vulture populations. From 2000 onwards, however, EU sanitary legislation has progressively limited the abandonment of dead animals in the field resulting in a sudden reduction of food availability with unknown ecological and conservation consequences. Here, we examine the dietary response of a tandem of carrion eaters, the griffon vulture (Gyps fulvus) and the Egyptian vulture (Neophron percnopterus), showing different degrees of dietary specialization. Our results showed that after the reduction in numbers of supplementary feeding stations (vulture restaurants) the niche breadth of the griffon vulture has broadened and now includes significant amounts of wild rabbits (Oryctolagus cuniculus) and garbage. The diet of the Egyptian vulture, on the contrary, did not vary substantially. The diet overlap showed patterns probably conditioned by interspecific competition and the progressive exploitation of unpredictable carcasses. On a short-term scale, consequences for smaller scavengers could be negative due to the monopolization of resources by the dominant and much more abundant griffon vulture, however in the long-term all guild species would benefit from the exploitation of unpredictable carcasses, which could enhance the possibilities of coexistence.     

Evidence of connectivity between continental and differentiated insular populations in a highly mobile species

Aim Genetically differentiated insular populations are candidates for independent units for conservation. However, occasional immigration to reduced island populations may occur and potentially have important consequences in their future viability and evolutionary potential. In this study, we investigate the conservation implications of population structure and connectivity of insular and continental populations of a migratory raptor as determined using genetic tools and satellite tracking. Location Western European populations in the Iberian Peninsula and two insular populations in the Mediterranean Sea (Balearic Islands) and Atlantic Ocean (Canary Islands). Methods We genotyped 22 microsatellite loci in 96 Egyptian vultures (Neophron percnopterus) from the Iberian Peninsula, 36 from Menorca (Balearic archipelago) and 242 (85% of the current population) from Fuerteventura (Canary Islands). We analysed genetic variation to estimate structure, gene flow, genetic diversity, effective size and recent demographic history of the populations. Additionally, 19 vultures were marked with satellite transmitters to track their migration routes. Results Insular populations were genetically differentiated from those of the mainland. We detected immigration in the insular populations and within the continental counterpart. We found similar levels of genetic variability between the continent and the islands, and a bottleneck analysis indicated recent sharp population declines in both archipelagos but not on the continent. Main conclusions Our study provides evidence that, in spite of significant differentiation, insular populations of highly mobile species may remain connected with the mainland. Conservation programmes should take into account population connectivity and integrate differentiated units of management within complex units of conservation that can best maintain processes and potential for evolutionary change.     

A Global Review of Causes of Morbidity and Mortality in Free-Living Vultures

Vulture species worldwide play a key role in ecosystems as obligate scavengers, and several populations have had precipitous declines. Research on vulture health is critical to conservation efforts including free-living vultures and captive breeding programs, but is limited to date. In this systematic review, we determined the reported causes of free-living vulture species morbidity and mortality worldwide. The most commonly reported cause of mortality was from toxins (60%), especially lead and pesticides, followed by traumatic injury (49%), including collisions with urban infrastructure and gunshot. Neglected areas of research in free-living vulture health include infectious diseases (16%), endocrine and nutritional disorders (6%), and neoplasia (<?1%). Almost half of the studies included in the review were conducted in either Spain or the USA, with a paucity of studies conducted in South America and sub-Saharan Africa. The highest number of studies was on Griffon (Gyps fulvus) (24%) and Egyptian vultures (Neophron percnopterus) (19%), while half of all vulture species had five or fewer studies. Future investigations on free-living vulture health should focus on neglected areas of research, such as infectious diseases, and areas with gaps in the current literature, such as South America, sub-Saharan Africa, and under-studied vulture species.

     

Genetic diversity of wild Cymbidium goeringii (Orchidaceae) populations from Hubei based on Inter-simple sequence repeats analysis

Cymbidium goeringii is a diploid and nonrewarding, bumblebee-pollinated species, which is distributed in China, Japan and Korea Peninsula. This species is now highly endangered due to the mass collection and forest clearance in China. In the present study, we investigated the distribution of genetic variation within and between eleven populations of Cymbidium goeringii in central China by using Inter-simple sequence repeats (ISSR) markers. Eleven primers produced a total of 127 clear and reproducible bands of which 112 were polymorphic. High genetic diversity was detected in Cymbidium goeringii for both population level (P = 63.1%; He = 0.194 5) and species level (P = 88.2%; He = 0.262 8). A higher level of genetic differentiation was detected among populations (G _ST = 0.244 0, F _ST = 0.220 7) with Nei’s G _ST analysis and analysis of molecular variance (AMOVA), and no correlation was found between geographical and genetic distance. Genetic drift rather than gene flow played an important role in forming the present population structure of Cymbidium goeringii. Limited gene flow among populations and gene drift increase the extinction risk of local populations. Some conservation concerns are therefore discussed together with possible strategies for implementing in situ and ex situ conservation. __________ Translated from Biodiversity Science, 2006, 14(3): 250–257 [译自: 生物多样性] Equally contributed authors     

Genetic diversity of the great bustard in Iberia and Morocco: risks from current population fragmentation

We studied the genetic diversity of great bustards (Otis tarda) in Iberia and Morocco, the main stronghold of this globally endangered species. Samples were collected from 327 individuals covering most of the distribution range within the study area. Sequence variation in a 657 bp fragment of the mtDNA control region revealed 20 variable sites defining 22 haplotypes, two of them exclusive to Morocco. Genetic diversity showed marked regional differences (π = 0–0.53, h = 0–0.89). Multidimensional scaling analysis based on F _ST values showed a clear division between Morocco and the Iberian Peninsula, with no evidence of current gene flow between them. Our results suggest that Morocco, where few matrilines have persisted to present, was colonized from Iberia thousands of years ago. Last century reports suggest dispersal through Gibraltar, when the species was more abundant at both sides of the Strait but later population declines and the Strait’s barrier effect have favoured current genetic isolation. Within Iberia, only the most peripheral populations (Navarra, Aragón and Andalusia) differed significantly from the main ones in central Spain. The first two showed extremely low genetic diversity and are probably threatened by inbreeding depression. Diversity was higher in Andalusia, where three exclusive haplotypes were found, suggesting some degree of isolation from other populations. Andalusia and Morocco could be regarded as separate management units which hold a significant proportion of the current genetic diversity and thus deserve urgent conservation measures.     

Population genetic structure of island and mainland populations of the quokka, Setonix brachyurus (Macropodidae): a comparison of AFLP and microsatellite markers

Translocation and reintroduction are important tools for the conservation or recovery of species threatened with extinction in the wild. However, an understanding of the potential genetic consequences of mixing populations requires an understanding of the genetic variation within, and similarities among, donor and recipient populations. Genetic diversity was measured using two independent marker systems (microsatellites and AFLPs) for one island and four small remnant mainland populations of Setonix brachyurus, a threatened medium sized macropod restricted to fragmented habitat remnants and two off-shore islands in southwest Australia. Microsatellite diversity in the island population (R _s = 3.2, H _e = 71%) was similar to, or greater than, all mainland populations (R _s = 2.1–3.9, H _e = 34-71%). In contrast, AFLP diversity was significantly lower in the island population (PPL = 20.5; H _j = 0.118) compared to all mainland populations (mean PPL = 79.5–89.7; mean H _j = 0.23–0.29). Microsatellites differentiated all (mainland and island) populations from each other. However, AFLP only differentiated the island population from the mainland populations—all mainland populations were not significantly differentiated from each other for this marker. Given a known time since isolation of the island population from the mainland (6,000 years ago), and an overall more conservative rate of evolution of AFLP markers, our results are consistent with mainland populations fragmenting thousands of years ago (but <6,000 years), probably as a consequence of reduced rainfall and the constriction of the preferred mesic habitat of quokkas. Our results also support a recent history of severe population bottlenecks in mainland populations, and a long history of bottlenecks of the island population, but reflect a recent explosion in numbers since European occupation of the island. Our results indicate that translocation of island populations to supplement mainland populations would introduce genetically markedly differentiated, and possibly maladapted, individuals.     

Allozyme variation in the sympatric ferns Culcita macrocarpa and Woodwardia radicans at the northern extreme of their ranges

Isozymes were used to study genetic variation in the clonal ferns Culcita macrocarpa and Woodwardia radicans in the northwestern Iberian Peninsula, their northern distributional limit. Despite their high chromosome numbers, both species were isozymic diploids. In C. macrocarpa all 18 resolved loci were monomorphic, with the same allele in all ramets from all populations. In W. radicans only two of the 16 interpreted loci were polymorphic, with two alleles per locus; ramet-level genotypes showed Hardy-Weinberg equilibrium, indicating an intergametophytic mating system; the number of genets distinguished was 1–3 per population; and among-population variation was low (F _ST = 0.231), suggesting effective gene flow (i.e. spore exchange). More generally, the very low (W. radicans, H _T = 0.012) or zero (C. macrocarpa) genetic diversity detected in the present study may be due to genetic drift associated with the reduction of populations in the last glaciation, and to founder effects in the subsequent Holocene expansion.     

Genetic variation of polymorphic <Emphasis Type="Italic">NOS</Emphasis> STR locus in ten Indian population groups

Genotyping of 313 random individuals belonging to ten different population groups from three different states of India was performed for polymorphic pentanucleotide repeat present in the 5′ flanking region of nitric oxide synthase gene (NOS2A) to study the effect of geographical and linguistic affiliations on the genetic affinities among these groups. Likelihood ratio tests showed that all the ten populations for this locus were in Hardy-Weinberg equilibrium. Eleven different alleles ranging from 7 repeat to 17 repeats and 46 different genotypes were observed. The observed and the expected heterozygosity ranged from 0.72–0.94 and 0.84–0.89, respectively. The discriminating power of this locus is ≥ 0.86 and the polymorphism information content of this locus in ten population groups ranged from 0.80 to 0.85. High PIC, PD and PE value of this STR showed this marker to be informative and can be used for DNA typing and population studies. The eight populations from Kerala showed a lower G _ST value of 0.016 compared to the G _ST of ten populations (G _ST = 0.019), thereby showing that the populations from the same state showed higher genetic proximity probably due to linguistic and geographical proximity between them. The article was received from the author in the original.     

Genetic structure of wild populations of the endangered Desert Pupfish complex (Cyprinodontidae: Cyprinodon)

Data from 10 microsatellite DNA loci were used to describe the genetic structure of the two extant species (Cyprinodon macularius and C. eremus) of the endangered Desert Pupfish complex of southwestern United States and northwestern Mexico. Variation at microsatellite loci was significantly correlated (Mantel test) with that of previous mtDNA results, both for the complex and for the relatively wide-ranging C. macularius alone. Both species showed unusually high levels of microsatellite diversity for non-marine fish (H _e = 0.84–0.93; A_R = 11.9–17.0). There was evidence (R _ST > F _ST) that the two extant populations of C. eremus have been isolated sufficiently long for mutation to contribute significantly to genetic divergence, whereas divergence among the nine assayed populations of C. macularius could be attributed to genetic drift alone. Correspondingly, 10% of the diversity in C. eremus was attributable to differences between the two populations, whereas, for C. macularius, only 2.7% was attributable to among-population variation. Within C. macularius, a small (0.8%), but statistically significant, portion was attributable to differences between populations in the Salton Sea area and those on the lower Colorado River delta. The two populations of C. eremus and five groups of populations of C. macularius are recommended as management units for conservation genetics management of the two species.     

Maintenance of genetic diversity in Cordia africana Lam., a declining forest tree species in Ethiopia

We assessed genetic variation in a total of 22 populations of the tree species Cordia africana Lam. in Ethiopia and analyzed the country-wide impact of population history, forest disturbance and alteration of land use on extant intraspecific diversity. Amplified Fragment Length Polymorphisms (AFLPs) and chloroplast microsatellite markers were investigated. The analyses of the AFLP data revealed high diversity in all investigated populations: the percentage of polymorphic loci (PPL) ranged from 62.2% to 92.2% and Nei's gene diversity from 0.220 to 0.320 within the populations. The mean PPL and the mean diversity within populations were 85.7% and 0.287, respectively. The analysis of molecular variance revealed a moderate level of differentiation (Φ_ST = 0.07, p < 0.001) among the populations. The Mantel test proved a significant but low correlation (r = 0.31, p < 0.001) between the geographic distance and the genetic differentiation matrices. Only three different cpDNA haplotypes were observed; no more than two haplotypes were found in any population. The dominant haplotype with an overall frequency of 81% was observed in all populations. The level of differentiation among the populations was comparatively low at chloroplast DNA (G _ST = 0.18, R _ST, N _ST = 0.22). The observed patterns and levels of genetic variation within and among the populations indicate that efficient gene flow via pollen and seed is likely to be the main factor contributing to the maintenance of genetic diversity in natural and disturbed conditions.     

The population genetic structure of a common tropical damselfish on the Great Barrier Reef and eastern Papua New Guinea

Understanding patterns of connectivity in marine species is vital for the management and conservation of marine biodiversity. Here, the population genetic structure of a common and abundant tropical reef damselfish, Pomacentrus amboinensis, is reported. Using nine polymorphic microsatellite loci, the genetic structure at both small (i.e., around Lizard Island, Great Barrier Reef [GBR]) and large (GBR and Papua New Guinea [PNG]) spatial and temporal scales (2–1,600 km; 28 days– 6 years; n = 1,119) was analyzed. Temporal analyses found no evidence of genetic differentiation within or between Lizard Island recruitment pulses (R _ST = −0.001, P = 0.788), or corresponding established adult populations separated by 6 years of sampling (R _ST = 0.003, P = 0.116). The spatial analysis revealed that P. amboinensis populations are largely panmictic on the GBR and eastern PNG (R _ST = 0.001, P = 0.913), the only genetic discontinuity being between Kimbe Bay to the north of PNG and all populations south of PNG (R _ST = 0.077, P < 0.0001). Despite assumed high levels of self-recruitment based on previous tagging studies (15–60%), data presented here indicate that enough recruits are dispersing to impede the evolution of genetic structure over distances as great as 1,600 kms in this species. Data therefore indicate that the temporal genetic stability recorded here is maintained by high levels of gene flow.     

Population Genetic Structure of Two Columnar Cacti with a Patchy Distribution in Eastern Brazil

The genetic variability and population genetic structure of six populations of Praecereus euchlorus and Pilosocereus machrisii were investigated. The genetic variability in single populations of Pilosocereus vilaboensis, Pilosocereus aureispinus, and Facheiroa squamosa was also examined. All of these cacti species have a patchy geographic distribution in which they are restricted to small areas of xeric habitats in eastern Brazil. An analysis of genetic structure was used to gain insights into the historical mechanisms responsible for the patchy distribution of P. euchlorus and P. machrisii. High genetic variability was found at the populational level in all species (P=58.9–92.8%, A_p=2.34–3.33, H_e=0.266–0.401), and did not support our expectations of low variability based on the small population size. Substantial inbreeding was detected within populations (F_IS=0.370–0.623). In agreement with their insular distribution patterns, P. euchlorus and P. machrisii had a high genetic differentiation (F_ST=0.484 and F_ST=0.281, respectively), with no evidence of isolation by distance. Accordingly, estimates of gene flow (N_m) calculated from F_ST and private alleles were below the level of N_m=1 in P. machrisii and P. euchlorus. These results favored historical fragmentation as the mechanism responsible for the patchy distribution of these two species. The genetic distance between P. machrisii and P. vilaboensis was not compatible with their taxonomic distinction, indicating a possible local speciation event in this genus, or the occurrence of introgression events.     

Plastid DNA variation in highly fragmented populations of <Emphasis Type="Italic">Microbiota decussata</Emphasis> Kom. (Cupressaceae), an endemic to Sikhote Alin Mountains

Microbiota decussata Kom. (Cupressaceae) is a subalpine species endemic to the Sikhote Alin Mountains with populations scattered throughout the range. We used sequence data for four noncoding regions of chloroplast DNA to characterize the genetic diversity in populations sampled from different parts of M. decussata natural range. No variation was observed in the trnT–trnF region, whereas the trnH–psbA, trnS–trnfM, and trnS–trnG regions showed polymorphisms. At the species level, we found a low nucleotide diversity (π = 0.0009) and high haplotype diversity (h = 0.981) as well as high differentiation (Φ_ST = 0.420). N _ST and G _ST values suggested the existence of a phylogeographic structure in M. decussata. The observed patterns of diversity could be explained in part by ecological features of the species and its long-term persistence throughout the range with population expansion, successive fragmentation and isolation.     

Vulture flight behavior and implications for aircraft safety

Growing vulture populations represent increasing hazards to civil and military aircraft. To assess vulture flight behavior and activity patterns at the Marine Corps Air Station in Beaufort, South Carolina, we equipped 11 black vultures (Coragyps atratus) and 11 turkey vultures (Cathartes aura) with solar-powered Global Positioning System (GPS) satellite transmitters during a 2-year study (1 Oct 2006–30 Sep 2008). Turkey vultures had larger seasonal home ranges than did black vultures, and 2 turkey vultures made round-trips to Florida. Black vultures consistently spent less time in flight (8.4%) than did turkey vultures (18.9%), and black vultures flew at higher altitudes than did turkey vultures in all seasons except summer when altitudinal distributions (above ground level) did not differ. Although we recorded maximum altitudes of 1,578 m for black vultures and 1,378 for turkey vultures, most flights were low altitude. A matrix of vulture flight altitude versus time of day revealed that >60% of vulture flight activity occurred from 4 hr to 9 hr after sunrise at altitudes below 200 m. Continuation of aggressive harassment coupled with flexible training schedules to avoid times and altitudes of high vulture activity will decrease hazards to aircraft posed by these birds. © 2011 The Wildlife Society.     

Patterns of genetic diversity and differentiation in the tsetse fly <Emphasis Type="Italic">Glossina morsitans morsitans</Emphasis> Westwood populations in East and southern Africa

Genetic diversity and differentiation within and among nine G. morsitans morsitans populations from East and southern Africa was assessed by examining variation at seven microsatellite loci and a mitochondrial locus, cytochrome oxidase (COI). Mean COI diversity within populations was 0.63 ± 0.33 and 0.81 taken over all populations. Diversities averaged over microsatellite loci were high (mean number of alleles/locus ≥7.4; mean H _E ≥ 65%) in all populations. Diversities averaged across populations were greater in East Africa (mean number of alleles = 22 ± 2.6; mean h _e = 0.773 ± 0.033) than in southern Africa (mean number of alleles = 18.7 ± 4.0; mean h _e = 0.713 ± 0.072). Differentiation among all populations was highly significant (R _ST = 0.25, F _ST = 0.132). Nei’s G _ij statistics were 0.09 and 0.19 within regions for microsatellites and mitochondria, respectively; between regions, G _ij was 0.14 for microsatellites and 0.23 for mitochondria. G _ST among populations was 0.23 for microsatellite loci and 0.40 for mitochondria. The F, G and R statistics indicate highly restricted gene flow among G. m. morsitans populations separated over geographic scales of 12–917 km.     

The diversity of chloroplast microsatellite loci in Siberian fir (Abies sibirica Ledeb.) and two Far East fir species A. nephrolepis (Trautv.) Maxim. and A. sachalinensis Fr. Schmidt

The genetic variability in 29 populations of Abies sibirica, three of A. nephrolepis, and seven of A. sachalinensis was studied using SSR markers of chloroplast DNA. Among ten primer pairs examined, pairs Pt71936 and Pt30204 gave stable amplification and polymorphic products (with nine and fourteen alleles, respectively). Totally, 70 haplotypes were found, 43 in A. sibirica, 49 in A. sachalinensis, and 31 in A. nephrolepis. The highest values of genetic diversity parameters were observed in A. sachalinensis, and the lowest in A. sibirica. The Siberian fir differs from Far East species by the uneven multimodal frequency distributions of allele length in both cpSSR loci, which is explained by the presence of few separated from each other dominating haplotypes. This fact indicates that A. sibirica and the Far East species have different demographic histories. In A. sibirica, the proportion of diversity between populations in the total genetic diversity, calculated taking into account the differences between haplotypes (R _ST) was 8.34 and 4.42% without accounting for haplotypes differences (R _ST > G _ST, P = 0.01). The pairwise G _ST correlate significantly with geographic distances between the populations A. sibirica and with genetic distances D calculated from allozyme data. No such correlations were found with the R _ST parameter. The results of cpSSR variability analysis strongly support the conclusions inferred from allozyme data: several geographic groups of comparatively genetically close populations are identified, which may be explained by the history of colonization of the present-day Siberian fir range. Original Russian Text ? S.A. Semerikova, V.L. Semerikov, 2007, published in Genetika, 2007, Vol. 43, No. 12, pp. 1637–1646.     

European wildcat populations are subdivided into five main biogeographic groups: consequences of Pleistocene climate changes or recent anthropogenic fragmentation?

Extant populations of the European wildcat are fragmented across the continent, the likely consequence of recent extirpations due to habitat loss and over‐hunting. However, their underlying phylogeographic history has never been reconstructed. For testing the hypothesis that the European wildcat survived the Ice Age fragmented in Mediterranean refuges, we assayed the genetic variation at 31 microsatellites in 668 presumptive European wildcats sampled in 15 European countries. Moreover, to evaluate the extent of subspecies/population divergence and identify eventual wild × domestic cat hybrids, we genotyped 26 African wildcats from Sardinia and North Africa and 294 random‐bred domestic cats. Results of multivariate analyses and Bayesian clustering confirmed that the European wild and the domestic cats (plus the African wildcats) belong to two well‐differentiated clusters (average Ф_ST = 0.159, R_st  = 0.392, P > 0.001; Analysis of molecular variance [AMOVA]). We identified from c. 5% to 10% cryptic hybrids in southern and central European populations. In contrast, wild‐living cats in Hungary and Scotland showed deep signatures of genetic admixture and introgression with domestic cats. The European wildcats are subdivided into five main genetic clusters (average Ф_ST = 0.103, R_st  = 0.143, P > 0.001; AMOVA) corresponding to five biogeographic groups, respectively, distributed in the Iberian Peninsula, central Europe, central Germany, Italian Peninsula and the island of Sicily, and in north‐eastern Italy and northern Balkan regions (Dinaric Alps). Approximate Bayesian Computation simulations supported late Pleistocene–early Holocene population splittings (from c. 60 k to 10 k years ago), contemporary to the last Ice Age climatic changes. These results provide evidences for wildcat Mediterranean refuges in southwestern Europe, but the evolution history of eastern wildcat populations remains to be clarified. Historical genetic subdivisions suggest conservation strategies aimed at enhancing gene flow through the restoration of ecological corridors within each biogeographic units. Concomitantly, the risk of hybridization with free‐ranging domestic cats along corridor edges should be carefully monitored.     

Stocking may increase mitochondrial DNA diversity but fails to halt the decline of endangered Atlantic salmon populations

Over the last 50 years, Spanish Atlantic salmon (Salmo salar) populations have been in decline. In order to bolster these populations, rivers were stocked with fish of northern European origin during the period 1974–1996, probably also introducing the furunculosis-inducing pathogen, Aeromonas salmonicida. Here we assess the relative importance of processes influencing mitochondrial (mt)DNA variability in these populations from 1948 to 2002. Genetic material collected over this period from four rivers in northern Spain (Cantabria) was used to detect variability at the mtDNA ND1 gene. Before stocking, a single haplotype was found at high frequency (0.980). Following stocking, haplotype diversity (h) increased in all rivers (mean h before stocking was 0.041, and 0.245 afterwards). These increases were due principally to the dramatic increase in frequency of a previously very low frequency haplotype, reported at higher frequencies in northern European populations proximate to those used to stock Cantabrian rivers. Genetic structuring increased after stocking: among-river differentiation was low before stocking (1950s/1960s Φ _ST = –0.00296–0.00284), increasing considerably at the height of stocking (1980s Φ _ST = 0.18932) and decreasing post-stocking (1990s/2002 Φ _ST = 0.04934–0.03852). Gene flow from stocked fish therefore seems to have had a substantial role in increasing mtDNA variability. Additionally, we found significant differentiation between individuals that had probably died from infectious disease and apparently healthy, angled fish, suggesting a possible role for pathogen-driven selection of mtDNA variation. Our results suggest that stocking with non-native fish may increase genetic diversity in the short term, but may not reverse population declines.     

Vulture distribution and people perception of vultures in Pokhara Valley,Nepal

Due to an abundance and diversity of vultures, Nepal is one of the most important countries for vulture conservation. Within Nepal, the Pokhara Valley is especially significant. We examine the distribution of vultures within the Pokhara Valley by conducting counts at 11 potential feeding or roosting sites using point count method. We further surveyed people of the valley regarding their perception of vulture ecology and conservation, knowledge of diclofenac use within the valley, and burial of livestock carcasses. We detected eight species of vultures, four of which are currently threatened with extinction. White‐rumped vulture Gyps bengalensis, Egyptian vulture Nephron percnopterus, and Himalayan vulture G. himalayensis were the most abundant. Almost all respondents (98%) had sighted the vultures in the wild. Formally educated respondents reported seeing vultures’ slightly more than nonformally educated respondents. Fifty‐eight percent respondents suspected habitat loss was the major threat for the vulture population decline in Pokhara Valley, and 97% respondents were not aware of any diclofenac use. The knowledge of vultures in people with different age groups suggests a more awareness programs are needed for local people, especially those who carry out animal husbandry and provide livestock to the vulture restaurant.