Ex situ transportation of coral larvae for research,conservation, and aquaculture

Due to the lack of appropriate methods to transport high amounts of larvae ex situ over large distances, the availability of coral larvae was so far mainly limited to their place of origin. For a research project at Rotterdam Zoo, The Netherlands, we transported several thousand larvae of three broadcast spawners (Acropora tenuis, A. digitifera, Diploria strigosa) from the Indo Pacific and the Caribbean to Europe. Beside logistics and packing techniques, post-transport survival rates were mainly influenced by larvae density and transport duration. Our results indicate optimum survival rates of >90% at densities of 4 larvae ml⁻¹ when not exceeding a transportation time of 4 days. The ex situ transport of coral larvae over large distances might offer new possibilities for research, conservation, and aquaculture.     

Allocating individuals to avoid inbreeding in ex situ conservation plantations: so far, so good

The only cost-effective way to control inbreeding in ex situ forest tree plantations is often to allocate trees in such a way that the possibility of close relatives mating is small and, consequently, inbreeding does not increase too much over time. The classical permutated neighbourhood methods look for the configuration in which no ramets of the same genet are planted in the surroundings (neighbourhoods) of a particular tree but deny the influence of more distant trees. Another limitation of these methods is that they cannot incorporate any other genetic (e.g. kinship) or ecological (e.g. phenology) information. We have developed a new method based on the minimisation of the global probability of generating inbred offspring for the whole population. Improvements of this method from the classical ones are: (i) it takes into account all the trees (whether near or far) and not only the neighbours; (ii) different pollen dispersal functions can be implemented, fitting the particularities of each species and population; (iii) it allows for the integration of all available information about the genetic relationship between trees; and (iv) it is flexible allowing for particular crosses to be banned or encouraged. The novel method showed a better performance than classical ones both for simulated data and a case study under a broad range of scenarios. Magnitude of the benefit depends on the actual and assumed parameters for the pollen dispersal function and the relationship between trees, but even in the simple case where only clone identity is considered some advantage can be obtained by implementing the new algorithm.     

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

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

Microbial diversity: ex situ conservation of Indonesian microorganisms

This paper discusses the role of microbial resource collections in the ex situ conservation of microbial diversity and, in particular, the activities of a recently completed Darwin Initiative project to facilitate collection, identification and exploitation of Indonesian microbial diversity. This helped revitalize the collection of the Lembaga Ilmu Pengetahuea Indonesia (Indonesian Institute of Sciences), LIPI, Research and Development Centre for Biology, Bogor, Indonesia. It is suggested that a focus on microorganisms beneficial to man is essential to enable ex situ collections to be a useful tool in the conservation and sustainable utilization of microbial diversity. This task is enormous and far beyond the capabilities of one collection and its achievement will necessitate a network of collections.     

Assessing statistical power of SNPs for population structure and conservation studies

Single nucleotide polymorphisms (SNPs) have been proposed by some as the new frontier for population studies, and several papers have presented theoretical and empirical evidence reporting the advantages and limitations of SNPs. As a practical matter, however, it remains unclear how many SNP markers will be required or what the optimal characteristics of those markers should be in order to obtain sufficient statistical power to detect different levels of population differentiation. We use a hypothetical case to illustrate the process of designing a population genetics project, and present results from simulations that address several issues for maximizing statistical power to detect differentiation while minimizing the amount of effort in developing SNPs. Results indicate that (i) while ~30 SNPs should be sufficient to detect moderate (F_ST = 0.01) levels of differentiation, studies aimed at detecting demographic independence (e.g. F_ST < 0.005) may require 80 or more SNPs and large sample sizes; (ii) different SNP allele frequencies have little affect on power, and thus, selection of SNPs can be relatively unbiased; (iii) increasing the sample size has a strong effect on power, so that the number of loci can be minimized when sample number is known, and increasing sample size is almost always beneficial; and (iv) power is increased by including multiple SNPs within loci and inferring haplotypes, rather than trying to use only unlinked SNPs. This also has the practical benefit of reducing the SNP ascertainment effort, and may influence the decision of whether to seek SNPs in coding or noncoding regions.     

Molecular genetic analysis of a captive-breeding program: the vulnerable endemic Jamaican yellow boa

The endemic Jamaican boa (or “yellow boa”, Epicrates subflavus) is a vulnerable species of the Caribbean biodiversity hotspot whose natural populations greatly declined mainly due to predation by introduced species, human persecution, and habitat destruction. A captive breeding program was initiated in 1976 and rationalized in 2002 by the establishment of a European Endangered Species Program. During the last 30 years, more than 600 offspring, of which 80 are still alive today, have been produced and distributed among European host institutions and privates. Here, using nine nuclear microsatellite loci and a fragment of the mitochondrial cytochrome b gene, we (i) determine the natural population from which the founders originate, (ii) identify parental allocation errors and ambiguities in the studbook, and (iii) assess the genetic diversity and estimate levels of inbreeding of the current captive population based on loss of alleles, variance in reproductive success, and relatedness among individuals. Combining measures of relatedness derived from multilocus genotypes with practical parameters such as age of animals and localization of host institutions, we propose mating groups that would maximize genetic diversity in the captive population of the Jamaican boa. Our analyses provide guidance for a more efficient breeding program that, in turn, could be used as the starting point of a repatriation program to increase the probability of the species long-term survival. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

A study on population genetic structure of Oryza meyeriana (Zoll. et Mor. ex Steud.) Baill. from Yunnan and its in situ conservation significance

In order to determine genetic diversity of Oryza meyeriana (Zoll. et Mor. ex Steud. ) Baill., 12 enzyme systems encoded by 17 loci were electrophoretically analyzed in 164 individuals of seven populations from Simao Prefecture, Yunnan Province, China. In comparison with those seed plants with the same life history and breeding systems, as well as the other species in the genus Oryza, the species shows rather low levels of genetic diversity (A = 1.1, P = 8.0%, Ho = 0.004 and He = 0. 015) within populations and high genetic differentiation among populations. FST was up to 0. 649, suggesting that 64. 9% of total genetic variability exists among populations. Considering high genetic differentiation among populations from a limited geographic region, most of the populations of the species are worth being protected, and therefore, great natural protection regions should theoretically be established in which a great deal of populations should be involved for developing in situ conservation management. Meanwhile     

Genetic structure of brown trout (<Emphasis Type="Italic">Salmo trutta</Emphasis> L.) from the Hardangervidda mountain plateau (Norway) analyzed by microsatellite DNA: a basis for conservation guidelines

The Hardangervidda in southern Norway, the largest mountain plateau in Europe, has thousands of lakes and streams, mainly between 1000 and 1300 m above sea level, where brown trout is the only fish species. To describe the current genetic diversity of brown trout in this area, a total of 863 fish from 20 lakes were genotyped with eleven microsatellites. Most diversity is within lake populations, but diversity among geographical groups and populations within groups was significant, too. Neighbor-joining, principle coordinate analysis and Bayesian clustering show three major geographic groups in accordance with the river systems. Bias was caused by recent stocking in two lakes. Low/no genetic differentiation among some populations indicates that intermixing is common when lakes are well-connected, as was also shown by assignment test. We recommend preserving the genetic diversity of brown trout in this unique area by managing stocking in lake systems according to genetic structure.     

The re-stocking of captive-bred ruffed lemurs (Varecia variegata variegata) into the Betampona Reserve, Madagascar: methodology and recommendations

Since November 1997 the Madagascar Fauna Group has released 13 captive-bred black and white ruffed lemurs (Varecia variegata variegata) into the Betampona Reserve in eastern Madagascar. The release programme has three major aims: (1) to assess the ability of captive-bred V. v. variegata to adapt to life in their natural habitat; (2) to investigate the contribution that such a release programme can make to reinforcing the existing small wild population of V. v. variegata in Betampona; and (3) to contribute to the long-term protection and conservation of the reserve. Criteria for the selection of release candidates, veterinary screening and pre-release experience in naturalistic environments are described and discussed. Methods for post-release monitoring of health and behaviour are covered in detail. The importance of considering the social dynamics of the species involved is emphasised. The survival of five of the releasees, plus successful reproduction and integration with the wild population have led to the conclusion that the release was a success.     

Genetic structure of American black bears in the desert southwest of North America: conservation implications for recolonization

American black bears (Ursus americanus) have recolonized parts of their former range in the Trans-Pecos region of western Texas after a >40-year absence. Assessment of genetic variation, structuring, gene flow, and dispersal among bear populations along the borderlands of Mexico and Texas is important to gain a better understanding of recolonization by large carnivores. We evaluated aspects of genetic diversity and gene flow for 6 sampling areas of black bears in southwestern North America using genotypic data from 7 microsatellite loci. Our results indicated that genetic diversity generally was high in the metapopulation of black bears in northern Mexico and western Texas. The episodic gene flow occurring via desert corridors between populations in northern Mexico and those in western Texas has permitted the establishment of only moderate levels of genetic structuring. Bayesian clustering analyses and assignment testing depicted the presence of 3 subpopulations among our 6 sampling areas and attested to the generally panmictic nature of bear populations in the borderlands region. The potentially ephemeral nature of the small populations in western Texas and genotypic characteristics of bears recolonizing these habitats attest to the importance of linkages along this portion of the borderlands of the United States and Mexico to effectively conserve and manage the species in this part of its range.     

Integrating population genetics and conservation biology in the era of genomics

As one of the final activities of the ESF-CONGEN Networking programme, a conference entitled ‘Integrating Population Genetics and Conservation Biology’ was held at Trondheim, Norway, from 23 to 26 May 2009. Conference speakers and poster presenters gave a display of the state-of-the-art developments in the field of conservation genetics. Over the five-year running period of the successful ESF-CONGEN Networking programme, much progress has been made in theoretical approaches, basic research on inbreeding depression and other genetic processes associated with habitat fragmentation and conservation issues, and with applying principles of conservation genetics in the conservation of many species. Future perspectives were also discussed in the conference, and it was concluded that conservation genetics is evolving into conservation genomics, while at the same time basic and applied research on threatened species and populations from a population genetic point of view continues to be emphasized.     

Estimating population structure under nonequilibrium conditions in a conservation context: continent-wide population genetics of the giant Amazon river turtle, Podocnemis expansa (Chelonia; Podocnemididae)

Giant Amazon river turtles, Podocnemis expansa, are indigenous to the Amazon, Orinoco, and Essequibo River basins, and are distributed across nearly the entire width of the South American continent. Although once common, their large size, high fecundity, and gregarious nesting, made P. expansa especially vulnerable to over-harvesting for eggs and meat. Populations have been severely reduced or extirpated in many areas throughout its range, and the species is now regulated under Appendix II of the Convention on International Trade in Endangered Species. Here, we analyse data from mitochondrial DNA sequence and multiple nuclear microsatellite markers with an array of complementary analytical methods. Results show that concordance from multiple data sets and analyses can provide a strong signal of population genetic structure that can be used to guide management. The general lack of phylogeographic structure but large differences in allele and haplotype frequencies among river basins is consistent with fragmented populations and female natal-river homing. Overall, the DNA data show that P. expansa populations lack a long history of genetic differentiation, but that each major tributary currently forms a semi-isolated reproductive population and should be managed accordingly.     

Preservation of genetic diversity in a wild and captive population of a rapidly declining mammal,the Common hamster of the French Alsace region

The Common hamster (Cricetus cricetus) faced massive population declines throughout its western range margin. In France, relict populations remained in the Alsace region. By comparing allelic diversity using microsatellite analysis over a time span of 12 years we investigated if this population decline led to genetic erosion in a French relict population of the species. Genetic diversity was moderate but comparable to other populations from Western Europe. Interestingly, no decline of allelic variation was revealed between 1999 and 2012 in the study region (expected heterozygosity = 0.51 in 1999 and 0.5 in 2012, respectively), suggesting a sufficiently high effective population size of ∼500 (179–956 SD). While several alleles were lost in a captive breed maintained for restocking purposes in the region, expected heterozygosity was comparably high (=0.5). Our results show that genetic diversity has been effectively maintained in a relict population of French Common hamsters despite of massive range loss. We recommend the maintenance of intense in situ conservation effort, along with regular monitoring of genetic diversity and effective population size.     

Strategies for ex situ conservation of Centaurea cineraria subsp. circae (Asteraceae), an endemic plant from Lazio (Italy)

Centaurea cineraria subsp. circae is an endemic plant with a distribution area limited to Circeo mountain (Lazio, Italy), whose population was estimated in a very low number of individuals. The aim of this work was to investigate ex situ conservation strategies such as achene collection and in vitro plant propagation, which will permit to carry out restoration programmes. The test carried out on the achenes demonstrated that only 5.5% of them were morphologically healthy. Seed germination tests showed that seeds do not display dormancy and that germination does not require pre-treatments. The higher germination rate (67.5%) was observed under a photoperiod of 12/12 h (light/dark) and temperature regime +20/+10°C. The in vitro studies demonstrated that micropropagation, acclimatization and the transfer outdoors of C. cineraria subsp. circae are not particularly difficult: 74% of shoot explants in a Murashige and Skoog (MS) medium added with 0.5 mg/l benzylaminopurine and 2 mg/l kinetin formed multiple shoots; 100% of shoots rooted in the MS medium added with 0.5 mg/l indole-3-butyric acid and over 90% survived the acclimatization phase. After been transferred outdoors, the totality of in vitro-propagated plants bloomed and appeared morphologically indistinguishable from wild plants. Preliminary chemical analyses showed a similar profile for in vitro-propagated and wild plants.     

Genetic diversity of European cattle breeds highlights the conservation value of traditional unselected breeds with high effective population size

In times of rapid global and unforeseeable environmental changes, there is an urgent need for a sustainable cattle breeding policy, based on a global view. Most of the indigenous breeds are specialized in a particular habitat or production system but are rapidly disappearing. Thus, they represent an important resource to meet present and future breeding objectives. Based on 105 microsatellites, we obtained thorough information on genetic diversity and population structure of 16 cattle breeds that cover a geographical area from the domestication centre near Anatolia, through the Balkan and alpine regions, to the North-West of Europe. Breeds under strict artificial selection and indigenous breeds under traditional breeding schemes were included. The overall results showed that the genetic diversity is widespread in Buša breeds in the Anatolian and Balkan areas, when compared with the alpine and north-western European breeds. Our results reflect long-term evolutionary and short-term breeding events very well. The regular pattern of allele frequency distribution in the entire cattle population studied clearly suggests conservation of rare alleles by conservation of preferably unselected traditional breeds with large effective population sizes. From a global and long-term conservation genetics point of view, the native and highly variable breeds closer to the domestication centre could serve as valuable sources of genes for future needs, not only for cattle but also for other farm animals.     

Genetic evidence of hybridization between the critically endangered Cuban crocodile and the American crocodile: implications for population history and in situ/ex situ conservation

Inter-specific hybridization may be especially detrimental when one species is extremely rare and the other is abundant owing to the potential for genetic swamping. The Cuban crocodile (Crocodylus rhombifer) is a critically endangered island endemic largely restricted to Zapata Swamp, where it is sympatric with the widespread American crocodile (C. acutus). An on-island, C. rhombifer captive breeding program is underway with the goals of maintaining taxonomic integrity and providing a source of individuals for reintroduction, but its conservation value is limited by lack of genetic information. Here we collected mtDNA haplotypic and nuclear genotypic data from wild and captive C. rhombifer and C. acutus in Cuba to: (1) investigate the degree of inter-specific hybridization in natural (in situ) and captive (ex situ) populations; (2) quantify the extent, distribution and in situ representation of genetic variation ex situ; and (3) reconstruct founder relatedness to inform management. We found high levels of hybridization in the wild (49.1%) and captivity (16.1%), and additional evidence for a cryptic lineage of C. acutus in the Antilles. We detected marginally higher observed heterozygosity and allelic diversity ex situ relative to the wild population, with captive C. rhombifer exhibiting over twice the frequency of private alleles. Although mean relatedness was high in captivity, we identified 37 genetically important individuals that possessed individual mean kinship (MK) values lower than the population MK. Overall, these results will guide long-term conservation management of Cuban crocodiles for maintaining the genetic integrity and viability of this species of high global conservation value.     

Genetic assessment of population restorations of the critically endangered Silene hifacensis in the Iberian Peninsula

In order to preserve endangered plant populations and recover their evolutionary potential and ecological behavior, some restoration measures generally involve the reinforcement of the population size in existing natural populations or the reintroduction of new populations. Genetic monitoring of both natural and restored populations can provide an assessment of restoration protocol success in establishing populations that maintain levels of genetic diversity similar to those in natural populations. The highly threatened Spanish species Silene hifacensis (Caryophyllaceae) has only three natural reduced mainland populations in the Iberian Peninsula, following decline and extinction that occurred during the late 20th century. Preterit restoration strategies were essentially based on the implantation of new populations and reinforcement of certain existing populations using transplants mostly cultivated in greenhouses. In the present contribution, levels and patterns of genetic variability within natural and restored populations of Silene hifacensis were assessed using the molecular technique AFLP. Our results pointed out significant genetic diversity differences across the three existing natural populations though their population fragmentation and progressive loss of individuals have not had an impact on the global genetic diversity of this species. For restored populations, their levels of genetic diversity were similar and even higher than in natural populations. As a result, the past restoration protocols were successful in capturing similar and even higher levels of genetic diversity than those observed within natural pools. However, inbreeding processes have been detected for two restored populations. Finally, the main source of plant material for the long-time restored transplants appears to be the natural population of Cova de les Cendres. This study demonstrates, once again, how genetic markers are useful tools to be taken in consideration for endangered plant species conservation plans.     

Microsatellite analysis of the genetic structure of captive forest musk deer populations and its implication for conservation

Forest musk deer (Moschus berezovskii) are rare as a result of poaching for musk and habitat loss. Some captive populations of forest musk deer have been established for decades in China. However, little genetic information is available for conservation management. In this paper, genetic variations, population structures, and the genetic bottleneck hypothesis were examined using 11 microsatellite loci from captive populations in Miyalo, Jinfeng and Maerkang in Sichuan Province, China. Estimates of genetic variability revealed substantial genetic variation in the three populations. A total of 142 different alleles were observed in 121 forest musk deer and the effective number of alleles per locus varied from 6.76 to 12.95. The average values of observed heterozygosity, expected heterozygosity, and Nei's expected heterozygosity were 0.552, 0.899 and 0.894 respectively. The overall significant (P < 0.001) deficit of heterozygotes because of inbreeding within breeds amounted to 34.5%. The mean F_ST (P < 0.001) showed that approximately 90.2% of the genetic variation was within populations and 9.8% was across populations. The UPGMA diagram, based on Nei's unbiased genetic distance, indicated that the three populations were differentiated into two different groups and it agreed with their origin and history. Bottleneck tests indicated that all three populations have undergone a population bottleneck, suggesting a small effective population size. Acknowledging that the genetic structure of populations has crucial conservation implications, the present genetic information should be taken into account in management plans for the conservation of captive forest musk deer.