Bridging the Research Gap between Live Collections in Zoos and Preserved Collections in Natural History Museums.

Zoos and natural history museums are both collections-based institutions with important missions in biodiversity research and education. Animals in zoos are a repository and living record of the world's biodiversity, whereas natural history museums are a permanent historical record of snapshots of biodiversity in time. Surprisingly, despite significant overlap in institutional missions, formal partnerships between these institution types are infrequent. Life history information, pedigrees, and medical records maintained at zoos should be seen as complementary to historical records of morphology, genetics, and distribution kept at museums. Through examining both institution types, we synthesize the benefits and challenges of cross-institutional exchanges and propose actions to increase the dialog between zoos and museums. With a growing recognition of the importance of collections to the advancement of scientific research and discovery, a transformational impact could be made with long-term investments in connecting the institutions that are caretakers of living and preserved animals.

First natural breeding of the endangered dusky gopher frog (Lithobates sevosus) in captivity.

To date, all captive breeding of the dusky gopher frog, Lithobates sevosus, a federally listed endangered species, has been accomplished using in vitro fertilization (IVF). Here, we describe multievent and highly fecund captive reproduction of dusky gopher frogs driven solely by natural environmental factors. Six pairs of L. sevosus were kept in a 3.7 × 4.4 m2 outdoor enclosure designed to resemble their natural breeding habitat, which included a pool and three artificial burrows. Modifications to the enclosure that simulated temperatures and conditions within their natural range during winter were added in October and removed in late February. Following a warm, rainy period, five egg masses were laid between March 5 and 11, 2020. The number of oocytes per egg mass was 2300 ± 409 (range = 1341-3565), with the total across all five egg masses being 11,501. Of these oocytes, the hatching rate was 68.58 ± 10.05% (range = 37.53%-95.59%), with a total of 7887 successful hatchlings overall. Clutch sizes were similar to those in the wild and greater than those typically produced using IVF; thus, natural breeding can substantially increase the number of frogs available for reintroduction programs. Although assisted reproductive technologies such as IVF will continue to be useful for ensuring the success of strategic genetic pairings of captive L. sevosus, the new tool of nonassisted reproduction in specifically designed outdoor enclosures is an important advancement for the conservation and recovery of this endangered species.

Effects of age on sperm quality metrics in endangered Mississippi gopher frogs (Lithobates sevosus) from captive populations used for controlled propagation and reintroduction efforts.

A decline in sperm quality with age is a common prediction of senescence-based hypotheses and empirical studies. While widely studied across taxa, there is little known on the effect of ageing on sperm quality in amphibians, especially in captive populations used for controlled propagation and reintroduction efforts. Here, we investigated variation in sperm quality metrics (i.e., motility, concentration, and morphology) in the endangered Mississippi gopher frog (Lithobates sevosus) among males of three age categories using individuals from captive breeding populations housed at three different zoological institutions. Different aged males across the species expectant lifespan (1-9, 1-2, 3-4, and 8-9-year-old subcategories) were chosen in an attempt to identify an optimal breeding age relevant for captive breeding programs. Moreover, we explored and statistically controlled for potential differences in sperm quality which may be attributed to the type of induction hormones and source populations that differed among institutions. Results indicated that males of different ages did not differ in sperm motility or concentration. However, we did find that older males (8-9 years old) had significantly longer sperm than other age categories and younger males (1-2 years old) had significantly more atypical sperm than other age categories. Furthermore, we found no significant differences in any sperm quality metrics between the different induction hormones or source populations used at the different institutions. Within a captive breeding program, this information is especially valuable as our results indicate that males that have only recently sexually matured may not be ready to breed, while older males maintain sperm quality metrics presumably related to fertilization success.

Inbreeding depression in sperm quality in a critically endangered amphibian.

Small, isolated populations often experience increased inbreeding and decreased heterozygosity, which increases the potential risk of inbreeding depression. The relationship between inbreeding and sperm health is well-documented in a variety of taxa, but has yet to be explored in amphibians. The dusky gopher frog, Lithobates sevosus, is a critically endangered species with years of documented inbreeding and low genetic variability as a consequence of isolation and population size reduction. This study investigated the effects of inbreeding on sperm quality in captive L. sevosus using an outbred, sister species (Lithobates pipiens) as a standard for comparison. We found L. sevosus to have severely reduced sperm quality in terms of total motility, forward progressive motility, concentration, and viability. Additionally, we observed a significant, negative relationship between total sperm motility and mean kinship within captive-bred individuals. These data serve to enhance our understanding of the role inbreeding plays in amphibians, and to provide valuable insight into new risk factors declining amphibian populations may face.

Corrigendum to: Sperm output and body condition are maintained independently of hibernation in an endangered temperate amphibian.

Hibernation is an integral part of the life history of species living in seasonal environments. However, our knowledge about the link between hibernation and reproductive success in amphibians remains limited, which poses an obstacle for critical conservation efforts. To fill this gap, we quantified the effects of captive hibernation on sperm quality, sperm quantity and body condition in an endangered anuran, the Wyoming toad (Anaxyrus baxteri), and used naturally hibernated wild toads as a standard for comparison. We hypothesised that hibernation is essential for optimal sperm output but is detrimental to body condition. Sperm collection was performed using assisted reproductive technologies for both captive and wild toads. Contrary to our hypotheses, no differences were observed in sperm metrics (total number of cells, concentration, motility and viability) or in body condition across captive treatment groups (0, 30 or 60 days of hibernation). Moreover, no difference was found between sperm metrics of captive toads and wild toads. These unexpected findings suggest that hibernation may not be an essential process for spermiation in A. baxteri while using exogenous hormones, and illustrate the potential of temperate amphibians to adapt to varying environmental conditions during winter months.

Sperm output and body condition are maintained independently of hibernation in an endangered temperate amphibian.

Hibernation is an integral part of the life history of species living in seasonal environments. However, our knowledge about the link between hibernation and reproductive success in amphibians remains limited, which poses an obstacle for critical conservation efforts. To fill this gap, we quantified the effects of captive hibernation on sperm quality, sperm quantity and body condition in an endangered anuran, the Wyoming toad (Anaxyrus baxteri), and used naturally hibernated wild toads as a standard for comparison. We hypothesised that hibernation is essential for optimal sperm output but is detrimental to body condition. Sperm collection was performed using assisted reproductive technologies for both captive and wild toads. Contrary to our hypotheses, no differences were observed in sperm metrics (total number of cells, concentration, motility and viability) or in body condition across captive treatment groups (0, 30 or 60 days of hibernation). Moreover, no difference was found between sperm metrics of captive toads and wild toads. These unexpected findings suggest that hibernation may not be an essential process for spermiation in A. baxteri while using exogenous hormones, and illustrate the potential of temperate amphibians to adapt to varying environmental conditions during winter months.

Cryopreservation and hormonal induction of spermic urine in a novel species: The smooth-sided toad (Rhaebo guttatus).

Global amphibian declines have fueled an increased interest in amphibian assisted reproductive technologies. Within the genus Rhaebo, half of the species are experiencing decreasing population trends; however, insufficient information is available on many of these species' reproductive biology. Using the smooth-sided toad, Rhaebo guttatus, we present effective methods for collecting and cryopreserving an example of Rhaebo sperm. Specifically, our findings show that administering 10 IU/g body weight of hCG (human chorionic gonadotropin) yields the most motile and concentrated sperm and that cryopreserving spermic urine in a solution of 5% DMFA (N,N-Dimethylformamide) and 10% trehalose returns sperm with a 33 ±â€¯3% average post-thaw motility. These findings may represent an important step forward in developing techniques that can be safely applied to other, more vulnerable species within the Rhaebo genus.

Immune and stress physiology of two captively-housed tortoise species.

Ecoimmunology affords us the ability to better understand immunological processes through consideration of external factors, such as the thermal microenvironment. This consideration is imperative when examining the immunological processes of ectothermic organisms like reptiles. Reptiles uniquely rely heavily on their innate immune function but remain poorly understood in immunological studies. In this study, we examined innate immunity in two zoo-housed tortoise species, the Indian star tortoise (Geochelone elegans, Schoepff, 1795) and northern spider tortoise (Pyxis arachnoides brygooi, Vuillemin & Domergue, 1972). Bacterial killing assays (BKAs) were optimized and used to assess the monthly immunocompetence of these tortoises to three different bacteria: Escherichia coli, Salmonella enterica, and Staphylococcus aureus. We evaluated differences in blood biochemistry values (lactate and glucose) among months and species as well as fecal corticosterone (CORT) between species. Lastly, we examined the potential influences of individual thermal microenvironments on bactericidal ability. Both G. elegans and P. a. brygooi demonstrated immunocompetence against all bacterial challenges, but only bactericidal ability against E. coli varied over months. Optimal BKA serum dilutions, blood glucose levels, and fecal CORT concentrations differed between the two species. Finally, there was evidence that the thermal microenvironment influenced the tortoises' bactericidal ability against E. coli. Through use of nonmodel organisms, such as tortoises, we are given insight into the inner workings of innate immunity and a better understanding of the complexities of the vertebrate immune system.

Using dermal glucocorticoids to determine the effects of disease and environment on the critically endangered Wyoming toad.

Amphibian populations are declining worldwide, and increased exposure to environmental stressors, including global climate change and pathogens like Batrachochytrium dendrobatidis (Bd), may be contributing to this decline. Our goal was to use a novel dermal swabbing method to measure glucocorticoid (GC) hormones and investigate the relationship among disease and environmental conditions in the critically endangered Wyoming toad (Anaxyrus baxteri). Our objectives were to (i) validate the use of dermal swabs to measure GCs using an adrenocorticotropic hormone (ACTH) challenge on eight captive toads (4 ACTH: 2 M, 2F and 4 saline as a control: 2 M, 2F), (ii) investigate stress physiology and disease status of toads across six reintroduction sites and (iii) compare dermal cortisol between reintroduced and captive toads. Dermal cortisol peaked immediately after the ACTH and saline injections. Faecal GC metabolites (FGMs) were significantly higher one week after the ACTH injection compared with the week before. Saline-injected toads had no change in FGM over time. Toads were only found in three reintroduction sites and dermal cortisol was similar across sites; however, reintroduced toads had higher dermal cortisol in August compared with June and compared with captive individuals. Bd status did not influence dermal cortisol concentrations. Dermal and faecal hormonal metabolite analyses can be used to study amphibian stress physiology and learn how environmental conditions are impacting population success.

Signals of forest degradation in the demography of common Asian amphibians.

BACKGROUND: Lowland areas in tropical East and Southeast Asia have a long history of conversion from forestland to agricultural land, with many remaining forests being chronically degraded by wood cutting, livestock grazing, and burning. Wetland-breeding amphibians that have evolved in lowland forests in the region have adjusted to changes in habitat composition caused by humans' activities, and populations continue to persist. However, we have little understanding of the impacts of forest disturbance on these species beyond assessments of abundance and distribution, and species considered to be common and widespread have been largely neglected. METHODS: We examined body condition and sex ratios of toads (Duttaphrynus melanostictus), predation risk in treefrogs (2 Polypedates spp.), and growth and survival of leaf litter frogs (2 Microhyla spp.) in agricultural land, degraded forest, and intact forest in two study areas, Thailand and Hong Kong. RESULTS: Toad populations exhibited higher body condition and female-biased sex ratios in intact forest. Predation of treefrog embryos by flies was lower in intact and degraded forests than in agricultural land. Embryonic survival and larval growth and survival in leaf litter frogs were lower in intact forests than in agricultural land. Results for each study were similar between study areas. DISCUSSION: For three of five of these common amphibian species, we documented signals of forest loss and disturbance in their populations. Although these species occur in disturbed habitats, loss of forest cover continues to degrade aspects of their population demography. We urge conservation biologists to consider that populations of species appearing to be common, widespread, and tolerant of human disturbance may be eroding over time.