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Humans are biased in their preferences for animals, and this may impact the composition of zoological collections. We assessed which kinds of animals the public and zoo professionals want to preserve in zoos for the future and analyzed these lists for previously identified biases and agreement across surveyed groups. We also characterized agreement among the surveyed groups on the roles of zoos and the composition of animal collections. We surveyed people who live in the United States, members of the Association of Zoos and Aquariums (AZA), and AZA zoo and aquarium directors. There was agreement among surveyed groups on the roles of zoos, though some differences existed. Zoo professionals and the public generally agreed on the emphasis on various categories of animals for zoo collections, though some differences were noted. We found evidence of bias towards mammals, charismatic megafauna, and felids across all surveyed groups. Agreement was high between AZA members and directors and moderate between zoo professionals and the public. These results indicate that these groups are generally in agreement about the roles of zoos, how they should compose their animal collections, and what animal species, in particular, should be kept in zoos for the future, allowing zoos to compose their collections to maximize delivery on their mission goals as well as address the preferences of the public. [ABSTRACT FROM AUTHOR]

The post mortem exam is important in diagnosing and investigating wildlife diseases. It is even more important to monitor the population of species that are not easily observed in the wild, such as otters. Of the 13 otter species, 11 are endangered due to many factors, such as climate change, pollution, or urbanisation. The authors present a review of the patterns of mortality of free-ranging otters worldwide, aiming to improve the knowledge of otter mortality and the role of mortality studies in their conservation. The main cause of death in aquatic otters is vehicle collision; hence, sea otter deaths are associated with shark attacks and imprisonment in fishing gear. It is possible to conclude that humans and human activity play a significant role in the mortality of these animals. In the future, a more standardised analysis of otter carcasses is necessary to understand their threats and diseases and design significant measures to protect these species. [ABSTRACT FROM AUTHOR]

Attention to habitat dynamics in subtropical mid-mountain forest plant communities containing endangered vegetation is critical for understanding the responses of ecosystems to global climate change and for their effective conservation. This study examines the species composition, structure, and interspecies competition within endemic and endangered Abies ziyuanensis (Abies ziyuanensis L.K.Fu and S.L.Mo) communities in China, comparing undisturbed and anthropogenically disturbed conditions. The survey recorded a total of 71 plant species across 39 families and 60 genera. PERMANOVA analysis highlighted significant disparities in species composition between the two forest community conditions. Communities impacted by anthropogenic disturbances showed a higher diversity of shrub and herbaceous species compared to those that were undisturbed, coupled with a significant increase in the number of Abies ziyuanensis seedlings, suggesting a greater potential for self-renewal. Nonetheless, the distribution of diameter class structures in these two community conditions indicates a declining trend in population numbers. In undisturbed Abies ziyuanensis communities, the Weighted Hegyi Competition Index (WCI) for Abies ziyuanensis was 6.04, below the average WCI of 12.24 for all trees within these communities. In contrast, within communities affected by anthropogenic disturbances, the WCI for Abies ziyuanensis reached 7.76, higher than the average WCI of 7.43 for all trees, indicating that Abies ziyuanensis in disturbed communities face heightened competitive pressure compared to undisturbed settings. These findings underscore that previous anthropogenic disturbances have altered the community composition, competition dynamics, growth environment, and succession trends of Abies ziyuanensis communities. While these disturbances promote the regeneration of Abies ziyuanensis, they also reduce its current dominance as a target species. [ABSTRACT FROM AUTHOR]

Plant cryobanks play a significant role in modern science and breeding. They contribute to the recovery of lost species, the emergence of new plant varieties, and help preserve and explore the diversity of the plant world. The IPPRAS Cryobank collection is constantly supplemented with new samples, while, at the same time, the stored samples are being monitored. In order to test seed germination, seeds of Allium and Veratrum species were thawed. Rare Allium species seeds, such as A. nutans, A. schoenoprasum, and A. victorialis were stored in liquid nitrogen for 17, 19, and 30 years, respectively. Long-term cryopreservation decreased germination rates for A. nutans from 96.55 to 50.00%, for A. schoenoprasum from 72.00 to 62.75%, and for A. victorialis from 90.00 to 83.05%. Seeds of a rare medicinal species, Veratrum lobelianum, were stored in liquid nitrogen for 18 years; the seed germination rate during this storage period has been significantly decreased from 75.00 to 14.81%. V. nigrum seeds were also collected and frozen in liquid nitrogen for 3 days. Short-term cryopreservation did not result in a statistically significant change in germination rates (from 79.71 to 82.69%). The seeds of an endangered ornamental species, Cypripedium calceolus, were collected and kept frozen for 3 days. After cryopreservation, the seeds were planted on three different media, as follows: ½ MS, MS with 10% coconut milk, and BM1. On ½ MS medium, 24.98% seeds formed protocorms, while on MS medium with 10% coconut milk, this number was 10.02%, and on BM1 medium, it was 15.02%, respectively; however, after 2.5 months, all of the protocorms died. Thus, it appears that the existing protocol for seed cryopreservation of C. calceolus needs further improvement. The size, weight, and free water content (WC) of six previously cryopreserved Stipa species and three Allium species were measured. For all the Allium and Stipa species studied, we found no correlation between seed size, WC, and cryotolerance. We also found no correlation between the life form, which reflects the water requirement of the species, and cryotolerance. [ABSTRACT FROM AUTHOR]

Eriodictyon capitatum (Namaceae) is a narrowly distributed shrub endemic to western Santa Barbara County, where it is known from only 10 extant California Natural Diversity Database element occurrences (EOs). Owing to low numbers of plants in nature, a limited overall extent, and multiple current threats, E. capitatum is listed as Endangered under the Federal Endangered Species Act and as Rare under the California Native Plant Protection Act. In the present study, high-throughput DNA sequence data were analyzed to investigate genetic diversity within and among all accessible EOs; to determine the extent of genetic isolation among EOs; to examine clonality within EOs; and to examine the taxonomic circumscriptions of E. capitatum, E. altissimum, E. angustifolium, and E. californicum through phylogenomic analysis. Population genetic analyses of E. capitatum reveal a pattern of strong genetic differentiation by location/EO. The clonality assessment shows that certain small EOs may support relatively few multilocus genotypes. The phylogenomic analyses strongly support the present-day taxonomic circumscriptions of both E. altissimum and E. capitatum, showing them to be reciprocally monophyletic and sister with strong support. Taken together, these results paint a picture of an evolutionarily and morphologically distinct species known from relatively few, genetically isolated stations. [ABSTRACT FROM AUTHOR]

Simple Summary: The gut microbiome can provide valuable information on the ecology and health of wildlife. We profiled the gut microbiota of the southern greater glider, an endangered Australian marsupial that was heavily disturbed by the 2019–2020 Australian bushfire season. Geographic location was found to be a significant driver of gut microbial community diversity and structure. In addition, the wildfires were shown to shape some aspects of the gut microbiome. By establishing baseline microbiome data for the southern greater glider, we lay the foundation for the future monitoring of populations at risk of compromised health, as well as inform future conservation management decisions for this endangered species. Studying the gut microbiome can provide valuable insights into animal health and inform the conservation management of threatened wildlife. Gut microbiota play important roles in regulating mammalian host physiology, including digestion, energy metabolism and immunity. Dysbiosis can impair such physiological processes and compromise host health, so it is essential that the gut microbiome be considered in conservation planning. The southern greater glider (Petauroides volans) is an endangered arboreal marsupial that faced widespread habitat fragmentation and population declines following the 2019–2020 Australian bushfire season. This study details baseline data on the gut microbiome of this species. The V3–V4 region of the 16S rRNA gene was amplified from scats collected from individuals inhabiting burnt and unburnt sites across southeastern Australia and sequenced to determine bacterial community composition. Southern greater glider gut microbiomes were characterised by high relative abundances of Firmicutes and Bacteroidota, which is consistent with that reported for other marsupial herbivores. Significant differences in gut microbial diversity and community structure were detected among individuals from different geographic locations. Certain microbiota and functional orthologues were also found to be significantly differentially abundant between locations. The role of wildfire in shaping southern greater glider gut microbiomes was shown, with some significant differences in the diversity and abundance of microbiota detected between burnt and unburnt sites. Overall, this study details the first data on greater glider (Petauroides) gut microbiomes, laying the foundation for future studies to further explore relationships between microbial community structure, environmental stressors and host health. [ABSTRACT FROM AUTHOR]

Human society is currently facing a growing demand for forest resources, which causes overexploitation and endangers biodiversity. In this regard, it is important to be aware that 10% of trees across the world are used in traditional and modern medicine. South America has the greatest diversity, with 40% of tree species in the world. The aim of our review consists of the assessment of the state of the art of micropropagation through somatic embryogenesis of representative medicinal trees in South America and of targeting the tree species that should be prioritized for conservation plans. From a total of 23,631 tree species from Central and South America, 31 are extinct, 7047 are threatened, and 1434 are possibly threatened. In this sense, in order to conserve them, various strategies are applied both in situ and ex situ. The application of in vitro multiplication protocols represents effective ways both in conservation and in the sustainable use of resources in order to obtain secondary metabolites of interest. Somatic embryogenesis is a well-known method in woody multiplication. According to the VOSviewer analyses, very few studies were available concerning aspects of somatic embryogenesis in medicinal trees. From the 10 representative species selected in our study, somatic embryogenesis protocols were established for 3 species, only for conservation purpose, not for secondary metabolites production. The development of protocols focused on obtaining secondary metabolites of medicinal trees will allow for the obtainment of valuable plant material as a non-invasive alternative. [ABSTRACT FROM AUTHOR]

Simple Summary: In recent years, due to industry and market preferences, local animal breeds have been exposed to genetic erosion and extinction risk. Effective strategies for their recovery and conservation are needed. Gentile di Puglia (GdP) is an autochthonous sheep breed, typical of Southern Italy, with an aptitude for wool, meat, and milk production and considerable historical and cultural value. The development of a GdP gamete and embryo cryobank could help to support the numerical reimplementation of this population. Animal germplasm conservation is mainly performed through sperm cryopreservation, whereas strategies on the female side are developed in a limited way. In this study, the dual purpose of monitoring the reproductive efficiency in one pilot GdP farm in the Apulia region and setting up a cryopreservation protocol, by vitrification, of immature cumulus-oocyte complexes (COCs) recovered from pre-pubertal lambs, followed by an analysis of their in vitro maturation potential and bioenergetic-oxidative status, was pursued. The results indicated that traditional reproductive management leads to progressive offspring reduction and that ex situ biotechnological conservation strategies, through immature oocyte vitrification and in vitro maturation, can support in situ conservation, leading to in vitro embryo production and transfer. Gentile di Puglia (GdP) is an autochthonous sheep breed of Southern Italy included among ovine breeds threatened by genetic erosion and extinction risk, which have been given attention by local and international institutions, thus emphasizing the need for germplasm conservation actions. In the present study, two assisted reproduction approaches, finalized for GdP conservation, were performed: (1) on-farm reproductive efficiency evaluation, expressed as pregnancy rate (PR), twin pregnancy rate (tPR), and body condition score (BCS), for three consecutive breeding cycles and (2) pre-pubertal lambs' immature cumulus–oocyte complex (COC) retrieval, vitrification, in vitro maturation (IVM), and assessment of meiotic stage and bioenergetic-oxidative status compared with those of other Italian and European commercial breeds. PR and tPR were progressively reduced over time. In all clinical examination times, BCS was significantly lower in nonpregnant ewes compared with pregnant ones. Fresh GdP pre-pubertal lamb COCs achieved meiotic maturation and showed healthy bioenergetic–oxidative status after IVM. Vitrification reduced the oocyte maturation rate in all groups. However, mature oocytes retained their cytoplasmic maturity, expressed as a mitochondria distribution pattern and activity, indicating promising developmental competence. In conclusion, clinical- and biotechnological-assisted reproduction approaches can support conservation strategies of GdP and other local sheep breeds in Southern Italy. [ABSTRACT FROM AUTHOR]

Environmental DNA (eDNA) techniques have emerged as a cost-effective and non-invasive strategy for monitoring the distribution of endangered aquatic species. Despite their numerous advantages, operational uncertainty at each step of the process represents a significant challenge that can impact the reliability of results. Hence, the optimization of the eDNA analytic protocol is of utmost importance. In this study, the rare and endangered fish species Sichuan taimen was chosen as the target species to evaluate the concordance between different approaches (tissue, pond, and field water samples) for eDNA detection. The results showed that membrane filtration, collection of a 2-L water sample, and utilization of the DNeasy Tissue and Blood DNA extraction kit were suitable for the analysis of DNA in water samples. Furthermore, the mtDNA D-loop region demonstrated potential as a specific molecular marker for the precise identification of Sichuan taimen. Our results indicated that TaqMan fluorescence PCR can serve as the optimal detection method for reliable identification of target DNA even at extremely low concentrations in the field. This study established an optimized eDNA analysis protocol for Sichuan taimen detection, which will provide the basis for future resource investigations aimed at protection of this endangered aquatic species. [ABSTRACT FROM AUTHOR]

Simple Summary: Resolving the taxonomic ambiguity of a species is important for their long-term conservation. We used non-invasive DNA sampling and camera trapping to address the taxonomic ambiguity in musk deer species in the Western Himalayas, and species occurrence locations were used to model the species distribution to identify suitable habitats. The combined results confirm the presence of only Kashmir musk deer (KMD) in the Western Himalayas. During our survey, no other musk deer species were found in the study area. This finding suggests that the presence of other species, such as Himalayan musk deer and Alpine musk deer, were incorrectly reported in the past. The predicted distribution (6% of the total area) of Kashmir musk deer is a narrow belt of the Western Himalayas between 2500 and 4500 m. Based on this study, we recommend long-term monitoring and assessment of KMD throughout its distribution range for its conservation and management. Insufficient research has been conducted on musk deer species across their distribution range, primarily because of their elusive behaviour and the fact they occupy remote high-altitude habitats in the Himalayas above 2500 m. The available distribution records, primarily derived from ecological studies with limited photographic and indirect evidence, fail to provide comprehensive information on the species distribution. Consequently, uncertainties arise when attempting to determine the presence of specific taxonomic units of musk deer in the Western Himalayas. This lack of knowledge hampers species-oriented conservation efforts, as there need to be more species-specific initiatives focused on monitoring, protecting, and combatting the illegal poaching of musk deer for their valuable musk pods. We used transect surveys (220 trails), camera traps (255 cameras), non-invasive DNA sampling (40 samples), and geospatial modelling (279 occurrence records) to resolve the taxonomic ambiguity, and identify the suitable habitat of musk deer (Moschus spp.) in Uttarkashi District of Uttarakhand and the Lahaul–Pangi landscape of Himachal Pradesh. All the captured images and DNA-based identification results confirmed the presence of only Kashmir musk deer (KDM) (Moschus cupreus) in Uttarakhand and Himachal Pradesh. The results suggest that KMD inhabit a narrow range of suitable habitats (6.9%) of the entire Western Himalayas. Since all evidence indicates that only KMD are present in the Western Himalayas, we suggest that the presence of other species of musk deer (Alpine musk deer and Himalayan musk deer) was wrongly reported. Therefore, future conservation plans and management strategies must focus only on KMD in the Western Himalayas. [ABSTRACT FROM AUTHOR]

Simple Summary: Red pandas are a threatened species, and zoos worldwide are working to conserve the species through international captive breeding programs. However, information on the physiology of captive red pandas is limited, which is hampering efforts to increase reproductive success and meet captive breeding goals. This study measured concentrations of fecal glucocorticoid (fGCM), progestagen (fPM), and androgen (fAM) metabolites in relation to environmental and biological factors among captive red pandas housed at three Indian zoos. Data revealed that fGCM concentrations were influenced by social time together (−), visitor numbers (+), number of nests (+), frequency of feedings (−), log density (−), and enclosure area (+). The fPM results suggested that enclosure area negatively affected concentrations in females. By contrast, fAM in males was not affected by these factors. This study suggests that employing more frequent feeding schedules and controlling the number of visitors may have positive effects on the welfare of red pandas, while higher fGCM and lower fPM in larger enclosures might be related to the limited enrichment and hiding spaces in those areas. Animals in human care are affected by stressors that can ultimately reduce fitness. When reproduction is affected, endangered species conservation programs can be severely compromised. Thus, understanding factors related to stress and reproduction, and related hormones, is important to ensure captive breeding success. Red pandas (Ailurus fulgens) are endangered, and populations in the wild are threatened with extinction. A global captive breeding program has been launched to conserve the species with the goal of reintroduction. However, there is little information on how stressors impact physiological aspects of the species. This study measured fecal glucocorticoid (fGCM), progestagen (fPM), and androgen (fAM) metabolite concentrations in 12 female and 8 male red pandas at 3 zoos in India to determine predictors of adrenal and gonadal steroid activity, and the influence of fGCM on reproduction. Based on the generalized linear mixed model (GLMM), fGCM concentrations were positively correlated with the number of visitors, number of nests and enclosure areas, and negatively related to frequency of feedings, log density, and social time, while fPM concentrations were negatively associated with enclosure areas. A confounder for enclosure areas and number of nests was the fact that these spaces were relatively barren, with limited hiding spaces, compared to the smaller enclosures. By contrast, no significant relationships were found for fAM, perhaps due to the smaller sample size. A negative relationship between fGCM and fPM was observed, indicating increasing adrenal hormones may decrease reproductive function among female red pandas. Results suggest that zoo management should consider increasing feeding frequency, providing larger enclosures with more enrichment and more nests in larger spaces, and regulating visitor numbers to support good welfare and potentially improve reproductive fitness of red pandas in captivity. [ABSTRACT FROM AUTHOR]

The present review paper is an attempt to examine and provide an overview of the various conservation strategies and regulatory framework to protect endangered plants, including Chlorophytum tuberosum, popularly known as Safed Musli in the local language. C. tuberosum belongs to the family Liliaceae and is being used in the indigenous systems of medicine as a galactagogue, aphrodisiac, antitumor, immunomodulatory, antidiabetic, analgesic, anti-inflammatory, hypolipidemic, anti-ageing, antimicrobial, etc. This plant has great medicinal and commercial value and is part of the Biological Diversity Act, but due to a lack of effective conservation, it is on the verge of extinction because of natural and manmade reasons, such as loss of habitat, climate change, pollution, excessive harvesting, etc. The most valuable medicinal plants have great importance; hence, many conservation techniques are being employed to protect them. In furtherance to the conservation of such plant species, strategic efforts, in the form of laws and policies, are laid; however, existing legislative mechanisms and policy parameters are not sufficient to overcome the challenges of conservation of such plant species, including Safed Musli, hence, this plant has been considered as a critically endangered plant in India. It is pertinent to note that we do not have specific legislation enacted for the protection of plant species; however, efforts are being made to conserve it under various laws, such as the Forest Conservation Act, Biological Diversity Act 2002, and many other allied legislations. This basic legislation of the Biological Diversity Act also lacks focal attention on the conservation of endangered plant species. Moreover, decentralization of power and actual community participation in conservation practices are also missing. A cumulative effect of both scientific measures and legal mechanisms supported by community participation may produce better results in the conservation of plant species, including Safed Musli. The protection of rich sources and biological diversity is not being taken as seriously as it ought to be, hence, it is necessary to improve awareness and public participation in conservation techniques with effective legislation for the conservation of highly endangered plant species. [ABSTRACT FROM AUTHOR]

Simple Summary: The population of Indo-Pacific humpback dolphin (Sousa chinensis) endemic to the inshore and estuarine waters of western Taiwan was identified as a subspecies (Sousa chinensis ssp. taiwanensis) and designated as "Critically Endangered" by the International Union for Conservation of Nature (IUCN). This population is at a high risk of extirpation, faces being impacted by some major anthropogenic threats, and numbers fewer than 65 individuals. Through a combination of long-term observations and photographic monitoring of identifiable individuals and stranding responses, accurate identification and classification of human-induced injuries and skin lesions is an effective approach and is particularly important to the management of the declining population. The results showed that the prevalence of skin marks is higher or comparable to the previous studies on the coastal cetacean populations across the globe. As the total population comprises dozens of individuals, radical measures to conserve the population and reduce environmental hazards are considered indispensable. As long-lived apex predators, Indo-Pacific humpback dolphins (Sousa chinensis) are key indicators of marine coastal environmental health. Skin marks can be observed on dorsal body surfaces that are visible during mark–recapture studies that rely on photo-identification (photo-ID) methods. Skin mark prevalence may be an indicator of environmental or anthropogenic stressors in the ecosystem, which may lead to individual and/or population-level health concerns. The prevalence of marks of anthropogenic origin was assessed in the Critically Endangered S. chinensis taiwanensis population along the coasts of central Taiwan. Fifty, twenty-eight, and thirty-four individuals were identified in 2018, 2019, and 2021, respectively. At least one category of injuries was observed in 47 of 57 distinctive individuals (82%), and adults showed a higher prevalence of deep injuries than the other coloration stages. At least one category of skin lesion was observed in 33 of 57 distinctive individuals (58%), and high prevalence of skin lesions was found in mature individuals. Given the difficulty in taking direct observations, skin mark prevalence is proposed as a proxy for estimating habitat health and anthropogenic stressors upon S. chinensis taiwanensis. The moderate-to-high prevalence of skin marks in this study was designated as a warning of risks. This study provides important updated information for the assessment of the health and survival of this population. More effective management measures are urgently needed to reverse the observed population decline. [ABSTRACT FROM AUTHOR]

Simple Summary: The spectacled flying fox (Pteropus conspicillatus) is an endangered frugivorous megabat with a restricted habitat in the Wet Tropics World Heritage Area in far north Queensland, Australia. Its population is in decline, and as such, mortality events in the species require investigation and the development of mitigation strategies. Cleft palate syndrome, first observed in the spectacled flying fox population in 1998, has produced sporadic neonatal mortality events over the past two decades. This study presents the rudimentary morphological signs of cleft palate syndrome in spectacled flying fox neonates, presenting gross pathology of syndromic signs upon visual inspection and a histological examination of palate malformations. It examines syndrome incidence data from the past two decades and reviews recent flying fox research to develop hypotheses related to the causes of the syndrome and to develop future research focuses. This, and future studies, will provide a greater understanding of the risk factors associated with the syndrome to guide the development of mitigation strategies that can deliver conservation outcomes for an endangered species, while presenting a unique opportunity for a novel comparative study into syndromic cleft palate in mammalian species. Cleft palate syndrome, first observed in the spectacled flying fox population in 1998, has produced sporadic neonatal mortality events over the past two decades, with an estimated incidence of up to 1/1000 births per year. This study presents a rudimentary characterisation of the syndrome, presenting gross pathology of syndromic signs upon visual inspection, a histological examination of palate malformations, and syndrome incidence data representing the past two decades. The syndrome presents with a range of signs, primarily congenital palate malformations ranging from a pinhole cleft to a complete hard and soft palate deficit, resulting in the death or abandonment of neonates shortly after birth. The congenital palate malformations are often associated with claw deformities, wiry facial hair, and in some instances, muscle weakness and neurological signs. The natural occurrence of the lethal congenital orofacial birth defects in the spectacled flying fox presents a unique opportunity for the investigation of putative aetiologies, drawing parallels between bat and other mammalian cleft palate risk factors. Further syndrome investigation has the potential to deliver both biodiversity conservation and comparative veterinary and biomedical outcomes. [ABSTRACT FROM AUTHOR]

Identifying connectivity patterns among remnant bird populations and their relationships with land use practices and adjacent habitat fragments is key to implementing appropriate long-term management strategies for species conservation. The coastal scrub and dune vegetation complex of the northern Yucatan Peninsula is rich in endemisms and has been affected by human development, which threatens the survival of the Yucatan Wren (Campylorhynchus yucatanicus) population, an endemic bird species. To identify possible anthropogenic barriers to the connectivity of C. yucatanicus along 14 localities in the Yucatan (Mexico) coastal north, we explored the relationship between the species population's genetic variability at each sampled site and landscape structure using regression models, in addition to the relationship between genetic distance and landscape resistance. Seven nuclear microsatellite loci were used as genetic markers. Four genetic populations were highlighted by the clustering method implemented in the Geneland program. Human settlement and availability of adequate habitat were significantly related to genetic distance (Fst), suggesting limited connectivity among sites due to ongoing land use changes. We suggest changing the IUCN threat category of C. yucatanicus to endangered as we found a significant loss of genetic variability in addition to restricted distribution, small population, habitat degradation, and loss of connectivity. [ABSTRACT FROM AUTHOR]

Bamboo is a plant that may replace plastics and wood as a new green environmental protection material that can be developed. Ferrocalamus strictus is a very special and rare bamboo which is a plant species with extremely small populations (PSESP) endemic to Yunnan, China, and it is listed as a China key national first-class protected wild plant. To explore the effects of extreme climate and environmental changes on this PSESP, we conducted a case study on the response of F. strictus to climate under two extreme future emission scenarios. We combined 29 environmental variables, including the temperature, precipitation, altitude, slope, aspect, soil pH, soil organic carbon content, and mean UV-B radiation of the highest month, to analyze the important environmental factors limiting the distribution of F. strictus and identified the priority conservation areas (PCAs) of F. strictus. Under the two future extreme climate scenarios, the core potential suitable distribution area of F. strictus would decrease by 26%–34% in 2040, 44%–46% in 2060, and 23%–58% in 2080, respectively. Bio18 (precipitation in the warmest quarter) and Bio3 (isotherm) were the two key factors limiting the geographical distribution of F. strictus (the contribution rates were 40.1% and 18.45%, respectively). The results of this study show that F. strictus is in urgent need of protection and PSESPs require more attention. [ABSTRACT FROM AUTHOR]

Simple Summary: Induced pluripotent stem cell (iPSC) technology is an emerging technique to reprogram somatic cells into iPSCs that have revolutionary benefits in the fields of drug discovery, cellular therapy, and personalized medicine. However, these applications are just the tip of the iceberg. There are examples now of repurposing iPSC technology to potentially aid endangered species, as well as even reviving extinct species. With increasing consumer reliance on animal products, combined with an exponentially growing population, there is a necessity to develop alternative approaches to conventional farming practices. One such approach that offers significant benefits is the development of domestic animal iPSCs, which will have a myriad of potential knock-on effects, such as reduction in animal death, pasture degradation, water consumption, and greenhouse gas emissions. Ultimately, reducing the environmental impact caused by large-scale farming provides an environmentally friendly commercial alternative. Another key issue that iPSC technology will address is both food security and potential zoonosis. Here, in the form of a "Frozen Ark", we will discuss past, current, and future research using iPSC technology and how it will potentially impact protein production in the form of animal-free meat alternatives to address increasing public awareness of foodborne diseases, factory farming, and the meat industry's ecological footprint. Induced pluripotent stem cell (iPSC) technology is an emerging technique to reprogram somatic cells into iPSCs that have revolutionary benefits in the fields of drug discovery, cellular therapy, and personalized medicine. However, these applications are just the tip of an iceberg. Recently, iPSC technology has been shown to be useful in not only conserving the endangered species, but also the revival of extinct species. With increasing consumer reliance on animal products, combined with an ever-growing population, there is a necessity to develop alternative approaches to conventional farming practices. One such approach involves the development of domestic farm animal iPSCs. This approach provides several benefits in the form of reduced animal death, pasture degradation, water consumption, and greenhouse gas emissions. Hence, it is essentially an environmentally-friendly alternative to conventional farming. Additionally, this approach ensures decreased zoonotic outbreaks and a constant food supply. Here, we discuss the iPSC technology in the form of a "Frozen Ark", along with its potential impact on spreading awareness of factory farming, foodborne disease, and the ecological footprint of the meat industry. [ABSTRACT FROM AUTHOR]

Bulbophyllum abangjoei is a new one-flowered orchid species described from Lanjak Entimau Wildlife Sanctuary, Katibas and Anap-Muput, Tatau, Sarawak. The taxonomic treatment is accompanied by taxonomic notes, information on the species' affinities at infrageneric level, geographical distribution, habitat ecology, and implications for their conservation. It is morphologically almost similar to Bulbophyllum deviantiae, B. membranifolium, and B. sanguineolentum, but clearly differs in the long, narrowly oblong to linear leaf blade, subsessile petiole ca. 5 mm long, long narrowly cylindrical pseudobulbs with some mature pseudobulbs reaching ¾ length of the leaf blade, non-resupinate flowers, shortly upcurved and not emarginate labellum apex, and elliptic-oblong stelids at lower margins near the apex of the column. For these morphological differences, B. abangjoei is here described as distinct and new species to science. The new species is so far only known from three localities in protected areas in Sarawak, and hereby is assessed as 'Endangered' [EN B2 (a)(b) D1] under IUCN Red List criteria B and D. [ABSTRACT FROM AUTHOR]

Parnassius bremeri (P. bremeri), a member of the genus Snow Apollo in the swallowtail family (Papilionidae), is a high alpine butterfly that lives in Russia, Korea, and China. It is an endangered wildlife (Class I) in South Korea and is a globally endangered species. The lack of transcriptomic and genomic resources of P. bremeri significantly hinders the study of its population genetics and conservation. The detailed information of the developmental stage-specific gene expression patterns of P. bremeri is of great demand for its conservation. However, the molecular mechanism underlying the metamorphic development of P. bremeri is still unknown. In the present study, the differentially expressed genes (DEGs) across the metamorphic developmental stages were compared using high-throughput transcriptome sequencing. We identified a total of 72,161 DEGs from eight comparisons. GO enrichment analysis showed that a range of DEGs were responsible for cuticle development and the melanin biosynthetic pathway during larval development. Pathway analysis suggested that the signaling pathways, such as the Wnt signaling pathway, hedgehog signaling pathway and Notch signaling pathway, are regulated during the developmental stages of P. bremeri. Furthermore, sensory receptors were also activated, especially during the larval to adult transition stage. Collectively, the results of this study provide a preliminary foundation and understanding of the molecular mechanism in their transcriptomes for further research on the metamorphic development of P. bremeri. [ABSTRACT FROM AUTHOR]

Simple Summary: Turtles are threatened all over the world. Malaysia has 24 species of turtles. This review focuses on current conservation status and some requirements for sustainability. We propose integrating concepts of ecology and molecular biology to provide almost comprehensive turtle reviews in Malaysia. Approximately 356 species of turtles inhabit saltwater and freshwater habitats globally, except in Antarctica. Twenty-four species of turtles have been reported in Malaysia, four of which are sea turtles. The state of Terengganu harbored the highest number of turtles, with 17 different reported species. Based on the IUCN Red List, 29% of turtle species in Malaysia are critically endangered. In comparison, another 25% are classified as endangered. Likewise, CITES reported that 67% of Malaysia's turtles are threatened, while 25% are classified as critically endangered. This review discusses the checklists, molecular genetics work, conservation status, recent trends, and recommendations for future research. Factors contributing to their population declines and current endangered status are also discussed. [ABSTRACT FROM AUTHOR]

Walker's Manihot, Manihot walkerae, is an endangered species endemic to south Texas and northeastern Mexico and is a Crop Wild Relative (CWR) of the international and economically important crop cassava (M. esculenta). Manihot walkerae is globally endangered (IUCN's Redlist, Texas list, USA); however, it is not recognized on the Mexican list of endangered species (NOM-059-SEMARNAT). We assessed the status of M. walkerae in Mexico and re-evaluated its global status. According to our analysis, M. walkerae should be considered an endangered species based on the IUCN's assessment method and a threatened species in Mexico based on the Mexican criteria. Our findings encourage the establishment of sound conservation plans for M. walkerae along the Texas–Mexico border. [ABSTRACT FROM AUTHOR]

The Interior Wetbelt (IWB) of British Columbia, which includes the globally rare Inland Temperate Rainforest (ITR), contains primary forests poorly attributed and neglected in conservation planning. We evaluated the IWB and ITR using four IUCN Red List of Ecosystems Criteria: geographic distribution, environmental degradation (abiotic and biotic factors), and likelihood of ecosystem collapse. Clearcut logging (3.2M ha) represented 57% of all anthropogenic disturbances, reducing potential primary forest by 2.7 million ha (28%) for the IWB and 524,003 ha (39%) for the ITR. Decadal logging rates nearly doubled from 5.3% to 10.2% from 1970s--2000s. Core areas (buffered by 100-m from roads and developments) declined by 70% to 95% for the IWB and ITR, respectively. Vulnerable was assigned to karst, the only abiotic factor assessed, because it was associated with rare plants. For biotic factors, Old-Growth Birds were Vulnerable, Southern Woodland Caribou (Rangifer tarandus caribou) habitat and Sensitive Fish were Endangered, and Old-Growth Lichens habitat was Critical. Overall, the IWB was ranked as Endangered and the ITR as Critical with core area collapse possible within 9 to 18 years for the ITR, considered one of the world's most imperiled temperate rainforests. [ABSTRACT FROM AUTHOR]

Simple Summary: The Andean cat is an endangered medium-sized felid with grey fur and a characteristic long tail with black rings. It is found only in cold and arid regions of the high Andes of Argentina, Bolivia, Chile, and Peru. As part of a collaboration between the Andean Cat Alliance (AGA) and Parque Andino Juncal (PAJ), we conducted a monitoring program from December 2020 to May 2021 using eight trail cameras in PAJ in the central Andes of Chile, an area where few records of the species have been described. We obtained records of Andean cats which fill a gap on the species distribution map, specifically in the area located between two previously identified evolutionary significant units (ESU) which are populations with different evolutionary histories that should be monitored, managed, and protected separately. Based on this new information, a comprehensive ESU reassessment across the range is currently being conducted by AGA. This will allow for a better understanding of the number of distinct populations across the range and the connectivity (i.e., current gene flow) among them, enabling the prioritization of small and isolated populations or units (i.e., higher extinction risk) and recommendations for evidence-based conservation strategies across the distribution. The Andean cat (Leopardus jacobita) is one of the most endangered and least known wild cat species in the Americas and the world in general. We describe new records of the Andean cat in the central Andes of Chile, in Parque Andino Juncal, obtained as part of a monitoring program conducted from December 2020 to May 2021 using eight trail cameras. The cameras were active for 135 days (sampling effort 1080 camera-trap days). We recorded Andean cats in two different cameras, corresponding to two independent events in January and March 2021, respectively (0.19% capture success). Our new records are relevant since they fill a gap on the species distribution map, specifically in the area located between the two previously identified evolutionarily significant units (ESU) (26–35° S) which has been prioritized by the Andean Cat Alliance (AGA). We highlight the relevance of private protected areas in ecosystems of high biodiversity and fragility such as Parque Andino Juncal and also of strategic private-social partnerships for successful collaborative efforts to monitor the presence of rare, elusive, and endangered species. Our next steps will be to collect scats from this newly identified site and conduct genetic analyses to determine whether these animals are part of previously identified ESUs or a yet unidentified potentially distinct ESU or MU requiring special conservation measures. [ABSTRACT FROM AUTHOR]

(1) Assessing growth and recruitment can be difficult during early life history, especially for rare species such as the pallid sturgeon Scaphirhynchus albus (federally endangered in the United States). One potential tool to address this knowledge gap is the stocking of age-0 individuals. (2) During July 2018, two stocking events of marked age-0 pallid sturgeon (53–56 or 41–42 days old) occurred in the lower Missouri River resulting in the release of 8495 individuals. Over the following three years, pallid sturgeon were sampled with baited trotlines and benthic trawls during routine monitoring. (3) During this period, we captured 77 individuals that recruited to age 1 or older. Only 9% of captured individuals yielded length-at-age values falling below the 95% prediction interval for pallid sturgeon raised entirely in an energetically favorable hatchery environment with abundant food resources. (4) These results suggest that the lower Missouri River is capable of supporting hatchery-level growth for young pallid sturgeon. This study also provided a unique opportunity to document recruitment to age-1 of age-0 pallid sturgeon stocked at relatively small sizes, which highlights the potential benefits of utilizing stocking to address endangered species knowledge gaps. [ABSTRACT FROM AUTHOR]

In 2018, the Minnesota Department of Natural Resources conducted the Sustainable Timber Harvest Analysis that resulted in the 10-year Stand Exam List (SEL). The SEL includes stands that will be assessed for potential management action from 2021 to 2030, but the location, number, and impact of stands actually harvested remains unknown. This study sought to use modeling to assess potential habitat changes from the SEL for five threatened, endangered, or special concern wildlife species. Three simulation scenarios captured the potential range of harvest from the SEL, and the Wildlife Habitat Indicator for Native Genera and Species model assessed associated habitat changes. The most realistic simulation scenario resulted in statistically insignificant habitat changes of less than ±6%, while two scenarios providing the upper and lower extremes of harvest resulted in statistically significant changes for one species each. Scenarios that resulted in less harvest and more mature forests benefited the five species, reflecting their habitat preferences. The tempering of habitat change values in the most realistic simulation scenario provides evidence for forest management tradeoffs between different wildlife species habitat requirements, as well as other forest resource management objectives. [ABSTRACT FROM AUTHOR]

An efficient procedure for in vitro shoot production of the medicinal plant sundew (Drosera rotundifolia L.) was developed. Of three cytokinins tested, zeatin (ZEA) at a concentration of 2 μM resulted in the formation of large numbers of adventitious shoots on leaf explants. A larger size of the small shoots was achieved by combining a 2-weeks preculture with ZEA (step 1: shoot induction) with a 5-weeks main culture without plant growth regulators (step 2: shoot elongation). Liquid media were superior to semisolid media: An average of 27.4 shoots per leaf explant, and 53.3 shoots per shoot explant were achieved. The 2-step culture system with liquid media permits a comparatively cheap micropropagation of sundew as well as in vitro biomass production, with potential for scale-up. [ABSTRACT FROM AUTHOR]

Simple Summary: Western chimpanzees are currently listed as a Critically Endangered subspecies. Human encroachment has taken a toll on this great ape due to fragmented habitat and the exchange of pathogens. This epidemiological study investigated the occurrence and genetic diversity of intestinal and blood parasites in faecal samples from wild chimpanzees living in the Dindefelo Community Nature Reserve, Senegal. We paid special attention to potential human-driven sources of infection and transmission pathways. Potential diarrhoea-causing protist parasites (e.g., Cryptosporidium spp., Giardia duodenalis, Entamoeba histolytica) were detected at low infection rates (and densities) or absent, whereas commensals (Entamoeba dispar) or protist of uncertain pathogenicity (Blastocystis sp.) were far more abundant. We detected Sarcocystis spp. in chimpanzee faeces. Blood protist parasites such as Plasmodium spp. and Trypanosoma brucei spp. (the etiological agents of malaria and sleeping sickness, respectively, in humans) were also found at low prevalences, but microfilariae of the nematode Mansonella perstans were frequently found. Molecular analyses primarily revealed host-adapted species/genotypes and an apparent absence of gastrointestinal clinical manifestations in infected chimpanzees. Zoonotic events of still unknown frequency and directionality may have taken part between wild chimpanzees and humans sharing natural habitats and resources. Wild chimpanzee populations in West Africa (Pan troglodytes verus) have dramatically decreased as a direct consequence of anthropogenic activities and infectious diseases. Little information is currently available on the epidemiology, pathogenic significance, and zoonotic potential of protist species in wild chimpanzees. This study investigates the occurrence and genetic diversity of intestinal and blood protists as well as filariae in faecal samples (n = 234) from wild chimpanzees in the Dindefelo Community Nature Reserve, Senegal. PCR-based results revealed the presence of intestinal potential pathogens (Sarcocystis spp.: 11.5%; Giardia duodenalis: 2.1%; Cryptosporidium hominis: 0.9%), protist of uncertain pathogenicity (Blastocystis sp.: 5.6%), and commensal species (Entamoeba dispar: 18.4%; Troglodytella abrassarti: 5.6%). Entamoeba histolytica, Enterocytozoon bieneusi, and Balantioides coli were undetected. Blood protists including Plasmodium malariae (0.4%), Trypanosoma brucei (1.3%), and Mansonella perstans (9.8%) were also identified. Sanger sequencing analyses revealed host-adapted genetic variants within Blastocystis, but other parasitic pathogens (C. hominis, P. malariae, T. brucei, M. perstans) have zoonotic potential, suggesting that cross-species transmission between wild chimpanzees and humans is possible in areas where both species overlap. Additionally, we explored potential interactions between intestinal/blood protist species and seasonality and climate variables. Chimpanzees seem to play a more complex role on the epidemiology of pathogenic and commensal protist and nematode species than initially anticipated. [ABSTRACT FROM AUTHOR]

Simple Summary: Why might a species both be seriously threatened and pose an overpopulation problem in introduced locations? The aim of this review was to understand the keys to the adaptive success of the wild rabbit (Oryctolagus cuniculus) in order to establish its strengths and weaknesses for the management of this keystone species in Mediterranean ecosystems. This work highlights the need to create specific conservation programs for this species. The European wild rabbit (Oryctolagus cuniculus) plays an important ecological role in the ecosystems of the Iberian Peninsula. Recently, rabbit populations have drastically reduced, so the species is now considered endangered. However, in some places, this animal is considered a pest. This is the conservation paradox of the 21st century: the wild rabbit is both an invasive alien and an endangered native species. The authors of this review aimed to understand the keys to the adaptive success of European rabbits, addressing all aspects of their biology in order to provide the keys to the ecological management of this species. Aspects including nutrition, genetics, immunity interactions with the environment, behaviour, and conflict with human activities were reviewed. Ultimately, rabbits are resilient and adaptable. The main adaptations that explain the rabbit's adaptive success are its nutrition (wide adaptation to food and good nutritional use of caecotrophy), immune system (powerful and developed), and other aspects related to genetics and behaviour. Rabbits' relationship with humans has led them to colonise other places where they have become pests. Despite these adaptations, populations in native places have been drastically reduced in recent years. Since it serves as a bastion of the Mediterranean ecosystem, a specific conservation program for this species must be carried out. Therefore, a study of the rabbit's response to diseases and nutrition (especially protein), as well as the interaction between them, is of special interest. [ABSTRACT FROM AUTHOR]

Marine bird populations have been declining globally with the factors driving this decline not fully understood. Viral diseases, including those caused by poxviruses, are a concern for endangered seabird species. In this study we have characterised a novel avipoxvirus, tentatively designated albatrosspox virus (ALPV), isolated from a skin lesion of an endangered New Zealand northern royal albatross (Diomedea sanfordi). The ALPV genome was 351.9 kbp in length and contained 336 predicted genes, seven of which were determined to be unique. The highest number of genes (313) in the ALPV genome were homologs of those in shearwaterpox virus 2 (SWPV2), while a further 10 were homologs to canarypox virus (CNPV) and an additional six to shearwaterpox virus 1 (SWPV1). Phylogenetic analyses positioned the ALPV genome within a distinct subclade comprising recently isolated avipoxvirus genome sequences from shearwater, penguin and passerine bird species. This is the first reported genome sequence of ALPV from a northern royal albatross and will help to track the evolution of avipoxvirus infections in this endangered species. [ABSTRACT FROM AUTHOR]

The genus Silene L. is one of the largest genera in Caryophyllaceae, and is distributed in the Northern Hemisphere and South America. The endemic species Silene leucophylla and the near-endemic S. schimperiana are native to the Sinai Peninsula, Egypt. They have reduced population size and are endangered on national and international scales. These two species have typically been disregarded in most studies of the genus Silene. This research integrates the Scanning Electron Microscope (SEM), species micromorphology, and the phylogenetic analysis of four DNA markers: ITS, matK, rbcL and psb-A/trn-H. Trichomes were observed on the stem of Silene leucophylla, while the S. schimperiana has a glabrous stem. Irregular epicuticle platelets with sinuate margin were found in S. schimperiana. Oblong, bone-shaped, and irregularly arranged epidermal cells were present on the leaf of S. leucophylla, while Silene schimperiana leaf has "tetra-, penta-, hexa-, and polygonal" epidermal cells. Silene leucophylla and S. schimperiana have amphistomatic stomata. The Bayesian phylogenetic analysis of each marker individually or in combination represented the first phylogenetic study to reveal the generic and sectional classification of S. leucophylla and S. schimperiana. Two Silene complexes are proposed based on morphological and phylogenetic data. The Leucophylla complex was allied to section Siphonomorpha and the Schimperiana complex was related to section Sclerocalycinae. However, these two complexes need further investigation and more exhaustive sampling to infer their complex phylogenetic relationships. [ABSTRACT FROM AUTHOR]

Humans currently face an extraordinary period of plant biodiversity loss. One strategy to stem further losses involves the development of species-level recovery plans that guide conservation actions. Seeds represent an important component in the life history of plants and are crucial for conservation activities. Yet, most recovery plans contain meager seed biology information. We set out to examine seed functional traits and germination niche breadth of Linum arenicola seeds exposed to a range of thermal, photoperiodic, and salinity gradients to gain perspectives on the seed biology of this endangered species that may inform conservation decision making and assist recovery plan development. We found that fresh seeds possess non-deep physiological dormancy, which may be alleviated via a four-week dry after-ripening treatment. The germination response of non-dormant seeds is subsequently promoted by constant rather than alternating temperatures. The optimum germination temperature range is 20–22 °C. Non-dormant seeds do not possess an absolute light requirement for germination, but are sensitive to low levels of salinity (EC50 = 6.34 ppth NaCl). The narrow thermal and salinity germination niche breadths reported here suggest a specialized reproductive strategy that may require careful consideration when planning ex and in situ conservation activities. [ABSTRACT FROM AUTHOR]

Walker's Manihot, Manihot walkerae, is an endangered plant that is endemic to the Tamaulipan thornscrub ecoregion of extreme southern Texas and northeastern Mexico. M. walkerae populations are highly fragmented and are found on both protected public lands and private property. Habitat loss and competition by invasive species are the most detrimental threats for M. walkerae; however, the effect of climate change on M. walkerae's geographic distribution remains unexplored and could result in further range restrictions. Our objectives are to evaluate the potential effects of climate change on the distribution of M. walkerae and assess the usefulness of natural protected areas in future conservation. We predict current and future geographic distribution for M. walkerae (years 2050 and 2070) using three different general circulation models (CM3, CMIP5, and HADGEM) and two climate change scenarios (RCP 4.5 and 8.5). A total of nineteen spatially rarefied occurrences for M. walkerae and ten non-highly correlated bioclimatic variables were inputted to the maximum entropy algorithm (MaxEnt) to produce twenty replicates per scenario. The area under the curve (AUC) value for the consensus model was higher than 0.90 and the partial ROC value was higher than 1.80, indicating a high predictive ability. The potential reduction in geographic distribution for M. walkerae by the effect of climate change was variable throughout the models, but collectively they predict a restriction in distribution. The most severe reductions were 9% for the year 2050 with the CM3 model at an 8.5 RCP, and 14% for the year 2070 with the CMIP5 model at the 4.5 RCP. The future geographic distribution of M. walkerae was overlapped with protected lands in the U.S. and Mexico in order to identify areas that could be suitable for future conservation efforts. In the U.S. there are several protected areas that are potentially suitable for M. walkerae, whereas in Mexico no protected areas exist within M. walkerae suitable habitat. [ABSTRACT FROM AUTHOR]

Megafishes are important to people and ecosystems worldwide. These fishes attain a maximum body weight of ≥30 kg. Global population declines highlight the need for more information about megafishes' conservation status to inform management and conservation. The northern Cambodian Mekong River and its major tributaries are considered one of the last refugia for Mekong megafishes. We collected data on population abundance and body size trends for eight megafishes in this region to better understand their conservation statuses. Data were collected in June 2018 using a local ecological knowledge survey of 96 fishers in 12 villages. Fishers reported that, over 20 years, most megafishes changed from common to uncommon, rare, or locally extirpated. The most common and rarest species had mean last capture dates of 4.5 and 95 months before the survey, respectively. All species had declined greatly in body size. Maximum body weights reported by fishers ranged from 11–88% of their recorded maxima. Fishers identified 10 threats to megafishes, seven of which were types of illegal fishing. Electrofishing was the most prevalent. Results confirm that Mekong megafishes are severely endangered. Species Conservation Strategies should be developed and must address pervasive illegal fishing activities, alongside habitat degradation and blocked migrations, to recover declining populations. [ABSTRACT FROM AUTHOR]

Population surveying and monitoring are important for identifying conservation needs and tracking trends in populations, communities, and ecosystems over time and laying the groundwork for conservation management and policy decisions. If species or populations go undetected because of inadequate effort or sampling design, protection and management cannot be properly provided. Due to the widespread loss of populations, the Eastern Massasauga (a rattlesnake) was recently listed as a federally threatened species in the United States; it is also listed as threatened in Canada. Given its current conservation status, there is considerable interest at state and federal levels in determining how to best survey for Eastern Massasaugas to aid in management decisions. Using a 16-year dataset, we examined the relationships among environmental, temporal, area, management, and search effort factors on the detection probability of Eastern Massasaugas. We found that four abiotic parameters (solar irradiance, shaded air temperature, three-day maximum air temperature, and humidity) and three search parameters (effort per researcher, search area, and search time of day) influenced detection of Eastern Massasaugas. As the current biodiversity crisis continues, the cost-effective use of resources and scientific expertise will continue to increase in importance. We hope our results stimulate similar analyses in other taxa, which will be critical for designing and implementing regional survey and monitoring programs. [ABSTRACT FROM AUTHOR]

Advances in nondestructive genetic sampling techniques continue to offer new opportunities for studying organisms, particularly those of conservation concern where more traditional invasive sampling methods are often not available. As part of a proof-of-concept, we investigated the effectiveness of using the chorion from residual butterfly egg debris as a source of viable genetic material for analysis. Laboratory material from a captive breeding population of the federally endangered Miami blue butterfly (Cyclargus thomasi bethunebakeri) was used to test efficacy and refine the methodology. The resulting best practices were subsequently evaluated using field-collected material from extant north Florida populations of the at-risk frosted elfin butterfly (Callophyrs irus). Our results demonstrated that it is possible to extract DNA of sufficiently high quantity and quality for successful gene sequencing. We additionally describe a simple, low-cost, and reliable method of collecting and storing egg debris samples that can be consistently adopted for field or laboratory work as well as deployed with projects that have a larger geographic scope and/or involve citizen scientists. Potential limitations related to field sample collection are discussed as well as needs for future evaluation. [ABSTRACT FROM AUTHOR]

Sonneratia × hainanensis, a species once endemic to Hainan Island in China, is now endangered. China's State Forestry Administration lists this species as a wild plant species with an extremely small population. Field fixed-point investigations, artificial pollination, and laboratory experiments, as well as other methods, were applied to study the reproductive system and seed germination of S. × hainanensis to elucidate the reasons for the endangerment of this species. The results are as follows: (1) Outcrossing index, pollen-ovule ratio, and artificial pollination showed S. × hainanensis has a mixed mating system and mainly focuses on outcrossing with some self-compatibility. (2) Fruit and seed placement tests showed that the fruit predators on the ground were mainly Fiddler crab and squirrel, with the predation rates being 100%. The artificially spread seeds do not germinate under natural conditions. The mean seed destruction rate and remaining rate of were 82.5% and 17.5%. (3) Seeds need to germinate under ambient light conditions, with an optimal photoperiod of 12 h. Seed germination is extremely sensitive to low temperatures because of optimum temperatures from 30 °C to 40 °C. At an optimal temperature of 35 °C, the seeds germinate under salinities ranging from 0‰ to 7.5‰, with an optimal salinity of 2.5‰, which shows the sensitivity of seed germination to salinity, with low salinity promoting germination, whereas high salinity inhibits germination. These findings indicate that the limited regeneration of S. × hainanensis is caused by the following: (1) Pollen limitation and inbreeding recession caused by the extremely small population of S. × hainanensis. (2) Seeds near parent trees are susceptible not only to high fruit drop rate, but to high predation beneath the parent trees′ canopy as well. (3) Seed germination has weak adaptability to light, temperature, and salinity. [ABSTRACT FROM AUTHOR]