Ex situ Diet Influences the Bacterial Community Associated with the Skin of Red-Eyed Tree Frogs (Agalychnis callidryas)

Amphibians support symbiotic bacterial communities on their skin that protect against a range of infectious pathogens, including the amphibian chytrid fungus. The conditions under which amphibians are maintained in captivity (e.g. diet, substrate, enrichment) in ex situ conservation programmes may affect the composition of the bacterial community. In addition, ex situ amphibian populations may support different bacterial communities in comparison to in situ populations of the same species. This could have implications for the suitability of populations intended for reintroduction, as well as the success of probiotic bacterial inoculations intended to provide amphibians with a bacterial community that resists invasion by the chytrid fungus. We aimed to investigate the effect of a carotenoid-enriched diet on the culturable bacterial community associated with captive red-eyed tree frogs (Agalychnis callidryas) and make comparisons to bacteria isolated from a wild population from the Chiquibul Rainforest in Belize. We successfully showed carotenoid availability influences the overall community composition, species richness and abundance of the bacterial community associated with the skin of captive frogs, with A. callidryas fed a carotenoid-enriched diet supporting a greater species richness and abundance of bacteria than those fed a carotenoid-free diet. Our results suggest that availability of carotenoids in the diet of captive frogs is likely to be beneficial for the bacterial community associated with the skin. We also found wild A. callidryas hosted more than double the number of different bacterial species than captive frogs with very little commonality between species. This suggests frogs in captivity may support a reduced and diverged bacterial community in comparison to wild populations of the same species, which could have particular relevance for ex situ conservation projects.     

Parks or arks: where to conserve threatened mammals?

Growing deterministic and stochastic threats to many wild populations of large vertebrates have focused attention on the conservation significance of captive breeding and subsequent reintroduction. However, work on both gorillas and black rhinos questions this shift in emphasis. In these species, field-based conservation can be effective if properly supported and, although this is not cheap, per capita costs may still be considerably lower than for ex situ propagation in captivity. Here we attempt to broaden the scope of this debate by contrasting the breeding success and costs of in situ and captive programmes for a range of threatened mammals. Data are scarce, but we find that across nine large-bodied genera, in situ conservation achieves comparable rates of population growth to those seen in established captive breeding programmes. Moreover, comparing budgets of well-protected reserves with zoos' own estimates of maintenance costs and the costs of zoo adoption schemes, we find that per capita costs for effective in situ conservation are consistently lower than those of maintenance in captivity. Captive breeding may be more cost-effective for smaller-bodied taxa, and will often remain desirable for large mammals restricted to one or two vulnerable wild populations. However, our results, coupled with the fact that effective in situ conservation protects intact ecosystems rather than single species, lead us to suggest that zoos might maximize their contribution to large mammal conservation by investing where possible in well-managed field-based initiatives, rather than establishing additional ex situ breeding programmes.     

Adaptations to Captivity in the Butterfly Pieris brassicae (L.) and the Implications for Ex situ Conservation

Captive breeding has been suggested as a method of conserving many threatened vertebrates, and is increasingly being proposed as a valuable conservation strategy for invertebrates. Potential genetic problems associated with ex situ conservation are widely recognized, but a further issue has received less attention: the possibility that populations will undergo adaptation to the captive environment, rendering them less well adapted to survival in the wild. We investigated six traits related to dispersal and reproduction in a culture of the large white butterfly Pieris brassicae (L.), that had been captive for c. 100–150 generations, and in recently wild stock reared simultaneously in a common environment. Individuals in the captive culture were heavier, with smaller wings and lower wing aspect ratios. Females from the captive culture laid many more eggs in cage experiments, and had higher ovary mass at the time of peak egg production. These differences are consistent with adaptation to captive conditions. Over time, similar evolutionary changes may affect invertebrates reared in ex situ conservation programmes, decreasing the likelihood that these species can be re-established in the wild. Although the timescale over which most vertebrates are likely to adapt to captivity is longer, and the traits involved will be different, invertebrates like P. brassicae may also provide a model of potential problems in long-term ex situ conservation programmes for both invertebrates and vertebrates. We suggest that measures to reduce or slow adaptation to captivity should be introduced alongside measures to reduce deleterious genetic effects in captive populations.     

Ex situ conservation genetics: a review of molecular studies on the genetic consequences of captive breeding programmes for endangered animal species

Captive breeding has become an important tool in species conservation programmes. Current management strategies for ex situ populations are based on theoretical models, which have mainly been tested in model species or assessed using studbook data. During recent years an increasing number of molecular genetic studies have been published on captive populations of several endangered species. However, a comprehensive analysis of these studies is still outstanding. Here, we present a review of the published literature on ex situ conservation genetics with a focus on molecular studies. We analysed 188 publications which either presented empirical studies using molecular markers (105), studbook analyses (26), theoretical work (38), or tested the genetic effects of management strategies using model species (19). The results show that inbreeding can be minimized by a thorough management of captive populations. There seems to be a minimum number of founders (15) and a minimum size of a captive population (100) necessary in order to minimize a loss of genetic diversity. Optimally, the founders should be unrelated and new founders should be integrated into the captive population successively. We recommend that genetic analyses should generally precede and accompany ex situ conservation projects in order to avoid inbreeding and outbreeding depression. Furthermore, many of the published studies do not provide all the relevant parameters (founder size, captive population size, H_o, H_e, inbreeding coefficients). We, therefore, propose that a general standard for the presentation of genetic studies should be established, which would allow integration of the data into a global database.     

Amphibians: suitable candidates for breeding-release programmes

Zoological parks are playing an increasingly important role in the management of threatened species through education, the maintenance of captive gene banks and the reintroduction of captive-bred animals into the wild. This paper discusses the suitability of amphibian species for reintroduction. The important features of amphibians for these programmes are highlighted, i.e., their high fecundity to allow rapid build-up of captive populations coupled with few behavioural problems with captive-bred animals, as well as the low cost of maintenance. The potential problems of inbreeding, poor retention of innate behavioural repertoires after multiple generation captive breeding, and the introduction of alien pathogens into wild populations are considered. Release programmes for two species, the Mallorcan midwife toad Alytes muletensis and the Puerto Rican crested toad Peltophryne lemur, are reviewed.     

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.     

Population viability analysis predicts decreasing genetic diversity in <Emphasis Type="Italic">ex situ</Emphasis> populations of the Itasenpara bitterling <Emphasis Type="Italic">Acheilognathus longipinnis</Emphasis> from the Kiso River,Japan

To conserve endangered species, the maintenance of ex situ captive populations with sustainable genetic diversity is often required, in combination with population viability analysis (PVA). Since 2010, the threatened Itasenpara bitterling Acheilognathus longipinnis lineages in the Kiso region, Japan, have been maintained in ex situ rearing facilities to allow for conservation efforts. In this study, we obtained microsatellite data from DNA extracted from these captive populations to elucidate their genetic diversity and effective population size. The populations of several initial generations indicated a deviation from Hardy–Weinberg equilibrium, probably due to the limited number of extracted founder individuals analyzed. The effective population size of the captive population tended to increase over the course of generations, although the degree of genetic diversity tended to decrease highlighting the concern for the progression of inbreeding. Our prediction based on the PVA suggests that the maintenance of the captive population under the current conditions could lead to extinction of the Itasenpara bitterling in 50 years. In contrast, simultaneously increasing the carrying capacity and individual exchange among populations appears to enhance the effective management of captive Itasenpara bitterling populations.     

Carry-over effect of captive breeding reduces reproductive fitness of wild-born descendants in the wild

Supplementation of wild populations with captive-bred organisms is a common practice for conservation of threatened wild populations. Yet it is largely unknown whether such programmes actually help population size recovery. While a negative genetic effect of captive breeding that decreases fitness of captive-bred organisms has been detected, there is no direct evidence for a carry-over effect of captive breeding in their wild-born descendants, which would drag down the fitness of the wild population in subsequent generations. In this study, we use genetic parentage assignments to reconstruct a pedigree and estimate reproductive fitness of the wild-born descendants of captive-bred parents in a supplemented population of steelhead trout (Oncorhynchus mykiss). The estimated fitness varied among years, but overall relative reproductive fitness was only 37 per cent in wild-born fish from two captive-bred parents and 87 per cent in those from one captive-bred and one wild parent (relative to those from two wild parents). Our results suggest a significant carry-over effect of captive breeding, which has negative influence on the size of the wild population in the generation after supplementation. In this population, the population fitness could have been 8 per cent higher if there was no carry-over effect during the study period.     

Enhancing the Conservation of Crop Wild Relatives in England

Humans require resilient, rapidly renewable and sustainable supplies of food and many other plant-derived supplies. However, the combined effects of climate change and population growth compromise the provision of these supplies particularly in respect to global food security. Crop wild relatives (CWR) contain higher genetic diversity than crops and harbour traits that can improve crop resilience and yield through plant breeding. However, in common with most countries, CWR are poorly conserved in England. There is currently no provision for long-term CWR conservation in situ, and comprehensive ex situ collection and storage of CWR is also lacking. However, there is a commitment to achieve their conservation in England’s Biodiversity Strategy and the UK has international commitments to do so as part of the Global Plant Conservation Strategy. Here, we identify a series of measures that could enhance the conservation of English CWR, thereby supporting the achievement of these national and international objectives. We provide an inventory of 148 priority English CWR, highlight hotspots of CWR diversity in sites including The Lizard Peninsula, the Dorset coast and Cambridgeshire and suggest appropriate sites for the establishment of a complementary network of genetic reserves. We also identify individual in situ and ex situ priorities for each English CWR. Based on these analyses, we make recommendations whose implementation could provide effective, long-term conservation of English CWR whilst facilitating their use in crop improvement.     

Rodrigues fruit bats (<Emphasis Type="Italic">Pteropus rodricensis</Emphasis>, Megachiroptera: Pteropodidae) retain genetic diversity despite population declines and founder events

Fruit bats of the genus Pteropus are important contributors to ecosystem maintenance on islands through their roles as pollinators and seed dispersers. However, island faunas are the most prone to extinction and there is a real need to assess the possible genetic implications of population reductions in terms of extinction risk. An effective method of ameliorating extinction risk in endangered species is the establishment of captive populations ex situ. The effectiveness of captive breeding programmes may be assessed by comparing the genetic variability of captive colonies to that of wild counterparts. Here, we use polymorphic microsatellite loci to assess genetic variability in wild, critically endangered Rodrigues fruit bats (Pteropus rodricensis, Dobson 1878) and we compare this variability to that in a captive colony. We document remarkable conservation of genetic variability in both the wild and captive populations, despite population declines and founder events. Our results demonstrate that the wild population has withstood the negative effects of population reductions and that captive breeding programmes can fulfil the goals of retaining genetic diversity and limiting inbreeding.     

The Genetic Integrity of the Ex Situ Population of the European Wildcat (Felis silvestris silvestris) Is Seriously Threatened by Introgression from Domestic Cats (Felis silvestris catus)

Studies on the genetic diversity and relatedness of zoo populations are crucial for implementing successful breeding programmes. The European wildcat, Felis s. silvestris, is subject to intensive conservation measures, including captive breeding and reintroduction. We here present the first systematic genetic analysis of the captive population of Felis s. silvestris in comparison with a natural wild population. We used microsatellites and mtDNA sequencing to assess genetic diversity, structure and integrity of the ex situ population. Our results show that the ex situ population of the European wildcat is highly structured and that it has a higher genetic diversity than the studied wild population. Some genetic clusters matched the breeding lines of certain zoos or groups of zoos that often exchanged individuals. Two mitochondrial haplotype groups were detected in the in situ populations, one of which was closely related to the most common haplotype found in domestic cats, suggesting past introgression in the wild. Although native haplotypes were also found in the captive population, the majority (68%) of captive individuals shared a common mtDNA haplotype with the domestic cat (Felis s. catus). Only six captive individuals (7.7%) were assigned as wildcats in the STRUCTURE analysis (at K = 2), two of which had domestic cat mtDNA haplotypes and only two captive individuals were assigned as purebred wildcats by NewHybrids. These results suggest that the high genetic diversity of the captive population has been caused by admixture with domestic cats. Therefore, the captive population cannot be recommended for further breeding and reintroduction.     

Population Genetics and the Effects of a Severe Bottleneck in an Ex Situ Population of Critically Endangered Hawaiian Tree Snails

As wild populations decline, ex situ propagation provides a potential bank of genetic diversity and a hedge against extinction. These programs are unlikely to succeed if captive populations do not recover from the severe bottleneck imposed when they are founded with a limited number of individuals from remnant populations. In small captive populations allelic richness may be lost due to genetic drift, leading to a decline in fitness. Wild populations of the Hawaiian tree snail Achatinella lila, a hermaphroditic snail with a long life history, have declined precipitously due to introduced predators and other human impacts. A captive population initially thrived after its founding with seven snails, exceeding 600 captive individuals in 2009, but drastically declined in the last five years. Measures of fitness were examined from 2,018 captive snails that died between 1998 and 2012, and compared with genotypic data for six microsatellite loci from a subset of these deceased snails (N = 335), as well as live captive snails (N = 198) and wild snails (N = 92). Surprisingly, the inbreeding coefficient (F_is) declined over time in the captive population, and is now approaching values observed in the 2013 wild population, despite a significant decrease in allelic richness. However, adult annual survival and fecundity significantly declined in the second generation. These measures of fitness were positively correlated with heterozygosity. Snails with higher measures of heterozygosity had more offspring, and third generation offspring with higher measures of heterozygosity were more likely to reach maturity. These results highlight the importance of maintaining genetic diversity in captive populations, particularly those initiated with a small number of individuals from wild remnant populations. Genetic rescue may allow for an increase in genetic diversity in the captive population, as measures of heterozygosity and rarified allelic richness were higher in wild tree snails.     

One Conservation: the integrated view of biodiversity conservation

The current global situation requires urgent decision-making to reverse processes of mass extinction of thousands of species. As a way of showing the importance of joint actions in this process, we aim to present the concept of One Conservation as a new proposal for the integration of sustainability, in situ and ex situ conservation for the restoration of ecosystems. According to the United Nations, we are beginning the decade of ecosystem restoration and in association with the International Union for Conservation of Nature guidelines, we can join efforts in the conservation of the planet. The survival of many species of wild animals depends on the management of populations currently maintained in ex situ conditions (under human care). To facilitate the exchange of genetic material between in situ and ex situ populations, reproductive biotechniques have become a great tool, making it possible to restore species in their natural environments. For effective conservation to occur, there must be an integrated view of the problem as a whole, and action for solutions must take place jointly by different spheres of society. Even more, conservation must be carried out by the public sector, the private sector, the third sector, and not less importantly, the agricultural sector. Therefore, One Conservation is defined as an interconnection between ex situ and in situ conservation plans, anthropic actions in the environment, and research in different areas that encompass conservation.     

Ultraviolet radiation and Vitamin D3 in amphibian health, behaviour, diet and conservation

Amphibians are currently suffering a period of mass extinction with approximately 20% of species under severe threat and more than 120 species already extinct. In light of this crisis there is an urgency to establish viable ex situ populations and also find the causes of in situ declines. The role of ultraviolet radiation and Vitamin D₃ in amphibian health directly influences both ex situ and in situ populations. Vitamin D₃ can be photosynthesised endogenously via UV-B radiation (UV-B), or acquired through the diet, and then metabolised to calcitriol the biologically active hormonal form. Although, there is a lack of literature concerning Vitamin D₃ requirements and calcitriol synthesis in amphibians, amphibians are likely to have similar Vitamin D₃ requirements and metabolic processes as other vertebrates due to the phylogenetically conservative nature of calcitriol biosynthesis. Deficiencies in calcitriol in amphibians result in nutritional metabolic bone disease (NMBD) and could compromise reproduction and immunity. However, excess biologically active UV radiation has also proven detrimental across all three amphibian life stages and therefore could impact both in situ and ex situ populations. Here we review the role and necessity of UV-B and calcitriol in amphibians and the potential for negative impacts due to excessive exposure to UV radiation. We also identify priorities for research that could provide critical information for maintaining healthy in ex situ and in situ populations of amphibians.     

Gap analyses to support ex situ conservation of genetic diversity in Magnolia, a flagship group

Worldwide about one third of all plant species is estimated to be threatened with extinction. Plants are generally under-represented in conservation. However, the global strategy for plant conservation (GSPC) sets forth 16 targets to halt the current and continuing loss of plant diversity within a framework for actions at global, regional, national and local levels. Target 8 of the GSPC directs that at least 75 % of threatened plant species be present in ex situ collections by 2020, but this target is far from being met. By analyzing where gaps in ex situ collections and research exist relative to diversity hotspots of threatened species, it is possible to identify and prioritize action. We undertake a gap analysis of ex situ collection needs in the genus Magnolia, a relatively well-known and valuable genus. Results indicate three main areas of biodiversity (China, Vietnam and Colombia) for the endangered Magnolia taxa. The present paper provides a critical overview of recent conservation activities carried out in the genus Magnolia throughout the world, focusing on genetic diversity analyses of the species.     

Conserving Plants in Gene Banks and Nature: Investigating Complementarity with Trifolium thompsonii Morton

A standard conservation strategy for plant genetic resources integrates in situ (on-farm or wild) and ex situ (gene or field bank) approaches. Gene bank managers collect ex situ accessions that represent a comprehensive snap shot of the genetic diversity of in situ populations at a given time and place. Although simple in theory, achieving complementary in situ and ex situ holdings is challenging. Using Trifolium thompsonii as a model insect-pollinated herbaceous perennial species, we used AFLP markers to compare genetic diversity and structure of ex situ accessions collected at two time periods (1995, 2004) from four locations, with their corresponding in situ populations sampled in 2009. Our goal was to assess the complementarity of the two approaches. We examined how gene flow, selection and genetic drift contributed to population change. Across locations, we found no difference in diversity between ex situ and in situ samples. One population showed a decline in genetic diversity over the 15 years studied. Population genetic differentiation among the four locations was significant, but weak. Association tests suggested infrequent, long distance gene flow. Selection and drift occurred, but differences due to spatial effects were three times as strong as differences attributed to temporal effects, and suggested recollection efforts could occur at intervals greater than fifteen years. An effective collecting strategy for insect pollinated herbaceous perennial species was to sample >150 plants, equalize maternal contribution, and sample along random transects with sufficient space between plants to minimize intrafamilial sampling. Quantifying genetic change between ex situ and in situ accessions allows genetic resource managers to validate ex situ collecting and maintenance protocols, develop appropriate recollection intervals, and provide an early detection mechanism for identifying problematic conditions that can be addressed to prevent further decline in vulnerable in situ populations.     

Exposure to Odors of Rivals Enhances Sexual Motivation in Male Giant Pandas

Males will alter their mating behavior to cope with the presence of their competitors. Even exposure to odors from potential competitors can greatly increase male ejaculate expenditure in a variety of animals including insects, fishes, birds and rodents. Major efforts have been made to examine males'' plastic responses to sperm competition and its fitness benefits. However, the effects of competitor absence on male''s sexual motivation and behaviors remain unclear, which has been proposed to be one of the causes for the poor sexual performance of some captive mammals. This study revealed that sexual motivation can be greatly enhanced in captive male giant pandas (Ailuropoda melanoleuca) by exposure to chemosensory cues from either one or three conspecifics males. It had been shown that potential rivals'' odors increased males'' chemosensory investigation behavior, as well as their observing, following and sniffing behaviors towards estrous females. Behaviors changed regardless of the number of rivals (one or three). Our results demonstrate the effects of potential competition on male giant pandas'' sexual motivation and behavioral coping strategy. We anticipate that our research will provide a fresh insight into the mechanisms underlying poor sexual performance in male captive mammals, and valuable information for the practical management and ex situ conservation of endangered species.     

Planning for optimal conservation of geographical genetic variability within species

Systematic Conservation Planning (SCP) involves a series of steps that should be accomplished to determine the most cost-effective way to invest in conservation action. Although SCP has been usually applied at the species level (or hierarchically higher), it is possible to use alleles from molecular analyses at the population level as basic units for analyses. Here we demonstrate how SCP procedures can be used to establish optimum strategies for in situ and ex situ conservation of a single species, using Dipteryx alata (a Fabaceae tree species widely distributed and endemics to Brazilian Cerrado) as a case study. Data for the analyses consisted in 52 alleles from eight microsatellite loci coded for a total of 644 individual trees sampled in 25 local populations throughout species’ geographic range. We found optimal solutions in which seven local populations are the smallest set of local populations of D. alata that should be conserved to represent the known genetic diversity. Combining these several solutions allowed estimating the relative importance of the local populations for conserving all known alleles, taking into account the current land-use patterns in the region. A germplasm collection for this species already exists, so we also used SCP approach to identify the smallest number of populations that should be further collected in the field to complement the existing collection, showing that only four local populations should be sampled for optimizing the species ex situ representation. The initial application of the SCP methods to genetic data showed here can be a useful starting point for methodological and conceptual improvements and may be a first important step towards a comprehensive and balanced quantitative definition of conservation goals, shedding light to new possibilities for in situ and ex situ designs within species.     

Genetic assessment of the Iberian wolf Canis lupus signatus captive breeding program

The main goal of ex situ conservation programs is to improve the chances of long term survival of natural populations by founding and managing captive colonies that can serve as a source of individuals for future reintroductions or to reinforce existing populations. The degree in which a captive breeding program has captured the genetic diversity existing in the source wild population has seldom been evaluated. In this study we evaluate the genetic diversity in wild and captive populations of the Iberian wolf, Canis lupus signatus, in order to assess how much genetic diversity is being preserved in the ongoing ex situ conservation program for this subspecies. A sample of domestic dogs was also included in the analysis for comparison. Seventy-four wolves and 135 dogs were genotyped at 13 unlinked microsatellite loci. The results show that genetic diversity in Iberian wolves is comparable in magnitude to that of other wild populations of gray wolf. Both the wild and the captive Iberian wolf populations have a similarly high genetic diversity indicating that no substantial loss of diversity has occurred in the captive-breeding program. The effective number of founders of the program was estimated as ∼ ∼16, suggesting that all founders in the studbook pedigree were genetically independent. Our results emphasize also the genetic divergence between wolves and domestic dogs and indicate that our set of 13 microsatellite loci provide a powerful diagnostic test to distinguish wolves, dogs and their hybrids.     

Coordinating conservation: global networking for species survival

The Species Survival Commission (SSC) of IUCN—the World Conservation Union is the largest, most comprehensive and diverse professional conservation network in existence today. It brings together over 6000 volunteers from more than 160 countries to contribute to the conservation of biological diversity by developing and executing programmes to study, save, restore, and manage wisely species and their habitats. The SSC is organized primarily along taxonomic lines, with members distributed in 96 Specialist Groups focusing on distinct groups of species. These groups are a source of the most current and expert information on the conservation status and priorities for action for species in the wild. As such, they provide an invaluable resource in setting priorities for captive propagation and in linking ex situ actions. In addition, the SSC has five disciplinary Specialist Groups that provide expertise to both the zoo community and field conservationists in areas such as reintroductions, veterinary medicine, captive breeding as a conservation tool, and the impact of invasive species on native flora and fauna. The effectiveness of the SSC is greatly enhanced by the close connections within its network between zoo professionals, academic scientists, field conservationists, and managers of natural resources. Challenges to the SSC currently include better information management, organization at the national level, and extending coverage in areas such as marine biodiversity.