The effects of preformed vitamin A and provitamin A carotenoid supplementation on tadpoles of the poison frog Phyllobates vittatus.

Understanding the nutritional requirements of captive animals is necessary for proper animal husbandry, however, the specific dietary requirements for many amphibian species commonly kept in captivity are unknown. Like most vertebrates, frogs cannot synthesize carotenoids and must therefore obtain these essential nutrients through diet. It is unclear if amphibians can cleave provitamin A carotenoids to form vitamin A metabolically within the body, so common practice is to supplement their captive diets with both preformed vitamin A and provitamin A carotenoids. We carried out a feeding experiment in tadpoles of Phyllobates vittatus, a commonly kept poison frog species, to test the effects of supplementing a fish flake diet with a provitamin A carotenoid (2.5 mg/g ß-carotene) and vitamin A (0.033-0.066 µg/mL retinyl acetate), both individually and in combination. Contrary to our expectations, supplementation had either no effect or adverse effects on tadpole growth and survivorship. Tadpoles reared under supplemented diets with vitamin A showed higher mortality rates, coupled with symptoms of hypervitaminosis A. Survivors had a smaller body size and mass at metamorphosis. ß-carotene supplementation alone had no detectable effect. The vitamin A and ß-carotene levels in our supplemented diet have been shown to be harmless or benefit tadpoles of other species, yet our results indicate that adding these amounts to what is found in a generalist fish flake mix can have detrimental effects on P. vittatus tadpoles. More broadly, this study highlights the importance of creating husbandry guidelines based on the specific physiological needs of the species (or species groups) being kept in captivity, rather than general ones for all amphibians, as is often done.

Tree frog-inspired nanopillar arrays for enhancement of adhesion and friction.

Bioinspired structure adhesives have received increasing interest for many applications, such as climbing robots and medical devices. Inspired by the closely packed keratin nanopillars on the toe pads of tree frogs, tightly arranged polycaprolactone nanorod arrays are prepared by mold process and chemical modification. Nanorod arrays show enhanced adhesion and friction on both smooth and rough surfaces compared to the arrays with hexagonal micropillars. The bonding of nanorods results in a larger stiffness of the nanorod surface, contributing mainly to friction rather than adhesion. The results suggest the function of closely packed keratin nanopillars on the toe pad of tree frogs and offer a guiding principle for the designing of new structured adhesives with strong attaching abilities.

Managing the Oocyte Meiotic Arrest-Lessons from Frogs and Jellyfish.

During oocyte development, meiosis arrests in prophase of the first division for a remarkably prolonged period firstly during oocyte growth, and then when awaiting the appropriate hormonal signals for egg release. This prophase arrest is finally unlocked when locally produced maturation initiation hormones (MIHs) trigger entry into M-phase. Here, we assess the current knowledge of the successive cellular and molecular mechanisms responsible for keeping meiotic progression on hold. We focus on two model organisms, the amphibian Xenopus laevis, and the hydrozoan jellyfish Clytia hemisphaerica. Conserved mechanisms govern the initial meiotic programme of the oocyte prior to oocyte growth and also, much later, the onset of mitotic divisions, via activation of two key kinase systems: Cdk1-Cyclin B/Gwl (MPF) for M-phase activation and Mos-MAPkinase to orchestrate polar body formation and cytostatic (CSF) arrest. In contrast, maintenance of the prophase state of the fully-grown oocyte is assured by highly specific mechanisms, reflecting enormous variation between species in MIHs, MIH receptors and their immediate downstream signalling response. Convergence of multiple signalling pathway components to promote MPF activation in some oocytes, including Xenopus, is likely a heritage of the complex evolutionary history of spawning regulation, but also helps ensure a robust and reliable mechanism for gamete production.

Effect of introducing frogs and fish on soil phosphorus availability dynamics and their relationship with rice yield in paddy fields.

The long-term impacts of introducing frogs and fish on rice yield and soil P availability are largely underestimated and undervalued. A 9-year field experiment compared the soil phosphorus fraction dynamics and their relationship with rice yield in rice-frog-fish (RFF) cultures, rice-fish (RF) cultures and rice-only (RO) cultures in southeastern China paddy fields. The yields in the RFF and RF cultures were notably higher than those in the RO culture, by 22.1% and 6.8%, respectively. Soil total P ranged from 345.5 to 385.6 mg kg-1 among all the farming systems, with the smallest amount found in the RO culture. There were only slight changes in the distribution of soil phosphorus fractions with time. The concentrations of NaHCO3-Pi and NaHCO3-Po were significantly higher in the RFF and RF cultures compared with those in the RO culture, and most of the NaOH-Pi and NaOH-Po fractions were greater in the RFF and RF cultures compared with those in the RO culture. The rice grain yield was significantly correlated with labile P and slowly cycling P. Introducing frogs and fish might be useful for increasing soil active P supplies and meeting rice nutrient requirements. This study concluded that RFF is the best practice for improving rice grain yields and soil fertility in paddy fields.

Detecting Amphibians in Agricultural Landscapes Using Environmental DNA Reveals the Importance of Wetland Condition.

Amphibians are declining worldwide, in part because of large-scale degradation of habitat from agriculture and pervasive pathogens. Yet a common North American amphibian, the wood frog (Lithobates sylvaticus), ranges widely and persists in agricultural landscapes. Conventional survey techniques rely on visual encounters and dip-netting efforts, but detectability limits the ability to test for the effects of environmental variables on amphibian habitat suitability. We used environmental DNA to determine the presence of wood frogs and an amphibian pathogen (ranavirus) in Prairie Pothole wetlands and investigated the effects of 32 water quality, wetland habitat, and landscape-level variables on frog presence at sites representing different degrees of agricultural intensity. Several wetland variables influenced wood frog presence, the most influential being those associated with wetland productivity (i.e., nutrients), vegetation buffer width, and proportion of the surrounding landscape that is comprised of other water bodies. Wood frog presence was positively associated with higher dissolved phosphorus (>0.4 mg/L), moderate dissolved nitrogen (0.1-0.2 mg/L), lower chlorophyll a (≤15 µg/L), wider vegetation buffers (≥10 m), and more water on the landscape (≥0.25). These results highlight the effects of environmental factors at multiple scales on the presence of amphibians in this highly modified landscape-namely the importance of maintaining wetland water quality, vegetation buffers, and surrounding habitat heterogeneity. Environ Toxicol Chem 2019;38:2750-2763. © 2019 SETAC.

Application of highly efficient and lowly toxic bufadienolides screened from toad skin in lymphatic chemotherapy for colorectal cancer through a lymphatic metastatic model.

BACKGROUND: Lymph node metastasis (LNM) remains a major obstacle to treat colorectal cancer (CRC). Increasing evidences have suggested that bufadienolides contain several fractions displaying antitumor activity and may be applied in lymphatic chemotherapy. However, effects of the highly efficient and lowly toxic (HELT) bufadienolides on CRC in lymphatic chemotherapy have not been reported. METHODS: Adenosine triphosphate tumor chemosensitivity assays (ATP-TCA) was performed to detect the inhibition rate (IR) of fractions of bufadienolides to cytokine-induced killer (CIK) cells and tumor cells. HELT fraction-loaded emulsions of different concentrations were prepared. Nude mouse bearing HCT116 tumors in footpad received high-dose emulsion (HD-E), middle-dose emulsion (MD-E), low-dose emulsion (LD-E), control emulsion (CE), Cinobufacini Injection (CI), or normal saline (NS), respectively. Hematoxylin and eosin (H&E) staining, Flow Cytometry (FCM), enzyme-linked immune sorbent assay (ELISA) and hematological examination were applied to evaluate therapeutic effects and potential toxicity. RESULTS: F18 and F19 were screened out as HELT fractions in vivo and F18-loaded emulsions of different concentrations for lymphatic administration were prepared. We confirmed that HD-E and MD-E produced obvious antitumor activities in footpad tumors and LNM compared with other groups in vitro. We also verified the effects of F18-loaded emulsions on activating hematopoietic function, stimulating proliferation of the spleen and natural killer (NK) cells, and promoting the secretion of IFN-γ and IgG1, although HD-E performed mild toxicity on liver. CONCLUSION: The present study demonstrated that lymphatic chemotherapy with HELT fraction of bufadienolides could be an effective approach to the treatment of CRC patients with LNM.

Comparison of the nutritional content of the captive and wild diets of the critically endangered mountain chicken frog (Leptodactylus fallax) to improve its captive husbandry.

It is vital to provide appropriate nutrition to maintain healthy populations in conservation breeding programs. Knowledge of the wild diet of a species can be used to inform captive diet formulation. The nutritional content of the wild diet of the critically endangered mountain chicken frog (Leptodactylus fallax) is unknown, like that of most amphibians. In this study, we analyzed the nutritional content of food items that comprise 91% of the wild diet of L. fallax, by dry weight of food items, and all food items offered to captive L. fallax at ZSL London Zoo and Jersey Zoo. We subsequently compared the nutritional content of the wild diet and captive diet at ZSL London Zoo consumed by L. fallax. To the authors' knowledge, this is the first study to directly compare the nutritional content of the wild and captive diets of an anuran amphibian. The captive diet at ZSL London Zoo, without dusting of nutritional supplements, was higher in gross energy and crude fat and lower in ash, calcium and calcium:phosphorus ratio than the wild diet. Most of the food items in the captive diets had a high omega-6:omega-3 fatty acid ratio and in the wild diet had a low omega-6:omega-3 fatty acid ratio. We recommend a combination of modifications to the captive diets to better reflect the nutritional content of the wild diet. Nutritional analysis of captive and wild diets is recommended for other species in conservation breeding programs to improve captive husbandry and ultimately fitness.

Effect of long-term dietary beta-carotene supplementation on sperm concentration and motility in an endangered amphibian.

Dietary carotenoids have a high antioxidant capacity, so it has been hypothesised that carotenoid supplimentation will improve sperm production and quality by protecting sperm from oxidative damage. The effects of carotenoids on sperm have only been assessed in three vertebrate species, and evidence for improved sperm concentration and motility remains equivocal. One reason for this might be that in most studies there has not been an assessment of the effects of single carotenoid compounds over a range of doses. Applied research focused on developing ways to improve sperm quality could benefit the captive breeding and conservation of threatened species. The aim of the present study was to assess a dose-dependent effect of beta-carotene supplementation on sperm concentration and motility in the endangered booroolong frog (Litoria booroolongensis). Individuals were supplemented with one of four beta-carotene doses (0, 0.1, 1 and 10 mg/g) from hatching until sexual maturity (53 weeks). Sperm concentration was determined prior to activation, and percent sperm motility and sperm velocity were measured at 0, 3 and 6 h post-activation using computer-assisted sperm analysis. Unexpectedly, beta-carotene had no significant effect on sperm concentration, percent motility or velocity at any time point, providing no evidence for beneficial effects. Findings of the present study indicate there are likely to be species-specific differences in sperm production and motility that influence the risk of oxidative damage to sperm and dependence on dietary antioxidants to inhibit these detrimental effects.

The effect of dietary antioxidants and exercise training on the escape performance of Southern Corroboree frogs.

Escape-response behaviour is essential to ensure an individual's survival during a predator attack, however, these behaviours are energetically costly and may cause oxidative stress. Oxidative stress can be reduced by supplementing an individual's diet with exogenous antioxidants or through regular moderate exercise training, which stimulates the upregulation of the endogenous antioxidant system. Two studies have tested the simultaneous effects of dietary antioxidant supplementation and exercise training on animal escape-response behaviour. The present study investigated the effects of dietary carotenoids and exercise training on the escape-response behaviour of Southern Corroboree frogs. Frogs were fed either a carotenoid-supplemented or unsupplemented diet and were exposed to repeated escape-response trials (training) for five consecutive weeks. Carotenoid-supplemented individuals outperformed unsupplemented individuals in initial hopping speed, length of the first hop and hopping distance, however, the performance of frogs in each treatment group became statistically similar after training. Within treatment groups, exercise training significantly improved the hopping speed of unsupplemented frogs, with speeds almost doubling between training weeks one and five. By contrast, exercise training did not significantly improve the hopping speed of carotenoid-supplemented frogs. Our results provide some of the first evidence that exercise training improves escape performance, and that dietary antioxidants may inhibit training-induced benefits.

The effect of antioxidants on sperm motility activation in the Booroolong frog.

Motile sperm can generate high levels of reactive oxygen species (ROS) post activation, and ROS can quickly accumulate to levels that impair motility and fertilising ability. The addition of antioxidants to sperm suspensions has been suggested as a means of reducing oxidative stress and enhancing sperm motility during and after sperm storage. Despite this, very few studies have attempted to experimentally test the effects of antioxidants on sperm motility activation in animals that use an external mode of fertilisation, espcially in amphibians. The present study aimed to investigate the effect of vitamin C and vitamin E on sperm motility activation in the Booroolong frog. Spermatozoa were activated in media containing either vitamin C (0, 0.05, 0.10, 0.15, 0.20, 0.25µgµL-1) or vitamin E (0, 0.25, 0.50, 0.75, 1.0, 1.25 1.50, 1.75µgµL-1). Sperm performance parameters (percent motility and velocity) were assessed using CASA at 0, 1, 2, 3, 4, 5 and 6h post-activation. Contrary to expectations, vitamin C supplementation was detrimental to sperm motility across all tested concentrations, while vitamin E had no effect. Further investigation on the endogenous antioxidant system of anuran sperm is required to ascertain whether alternative antioxidants may be more suitable at reducing ROS produced during sperm activation and improving sperm motility activation in vitro.

Testing the effect of dietary carotenoids on larval survival, growth and development in the critically endangered southern corroboree frog.

The success of captive breeding programs (CBPs) for threatened species is often limited due to a lack of knowledge of the nutritional conditions required for optimal growth and survival. Carotenoids are powerful antioxidants known to accelerate vertebrate growth and reduce mortality. However, the effect of carotenoids on amphibian life-history traits remains poorly understood. The aim of our study was to use a manipulative laboratory experiment to test the effect of dietary-carotenoid supplementation during the larval life stage on the survival, growth and development of the critically endangered southern corroboree frog (Pseudophryne corroboree). Larvae were fed either a carotenoid supplemented diet or an unsupplemented diet and the survival, growth and development of individuals was monitored and compared. There was no significant effect of dietary treatment on larval survival, growth rate, time taken to reach metamorphosis, or body size at metamorphosis. Our findings provide no evidence that carotenoid supplementation during the larval life stage improves the growth and development of southern corroboree frogs. However, because the carotenoid dose used in our study did not have any detrimental effects on P. corroboree larvae, but has previously been shown to improve adult coloration, immunity, and exercise performance, carotenoid supplementation should be considered when evaluating the nutritional requirements of P. corroboree in captivity. Carotenoid supplementation studies are now required for a diversity of anuran species to determine the effects of carotenoids on amphibian survival, growth and development. Understanding the effects of dietary carotenoids on different life-history traits may assist with amphibian captive breeding and conservation.

Animal models for auditory streaming.

Sounds in the natural environment need to be assigned to acoustic sources to evaluate complex auditory scenes. Separating sources will affect the analysis of auditory features of sounds. As the benefits of assigning sounds to specific sources accrue to all species communicating acoustically, the ability for auditory scene analysis is widespread among different animals. Animal studies allow for a deeper insight into the neuronal mechanisms underlying auditory scene analysis. Here, we will review the paradigms applied in the study of auditory scene analysis and streaming of sequential sounds in animal models. We will compare the psychophysical results from the animal studies to the evidence obtained in human psychophysics of auditory streaming, i.e. in a task commonly used for measuring the capability for auditory scene analysis. Furthermore, the neuronal correlates of auditory streaming will be reviewed in different animal models and the observations of the neurons' response measures will be related to perception. The across-species comparison will reveal whether similar demands in the analysis of acoustic scenes have resulted in similar perceptual and neuronal processing mechanisms in the wide range of species being capable of auditory scene analysis.This article is part of the themed issue 'Auditory and visual scene analysis'.

Breeding Guild Determines Frog Distributions in Response to Edge Effects and Habitat Conversion in the Brazil's Atlantic Forest.

Understanding the response of species with differing life-history traits to habitat edges and habitat conversion helps predict their likelihood of persistence across changing landscape. In Brazil's Atlantic Forest, we evaluated frog richness and abundance by breeding guild at four distances from the edge of a reserve: i) 200 m inside the forest, ii) 50 m inside the forest, iii) at the forest edge, and iv) 50 m inside three different converted habitats (coffee plantation, non-native Eucalyptus plantation, and abandoned pastures, hereafter matrix types). By sampling a dry and a wet season, we recorded 622 individual frogs representing 29 species, of which three were undescribed. Breeding guild (i.e. bromeliad, leaf-litter, and water-body breeders) was the most important variable explaining frog distributions in relation to edge effects and matrix types. Leaf-litter and bromeliad breeders decreased in richness and abundance from the forest interior toward the matrix habitats. Water-body breeders increased in richness toward the matrix and remained relatively stable in abundance across distances. Number of large trees (i.e. DBH > 15 cm) and bromeliads best explained frog richness and abundance across distances. Twenty species found in the interior of the forest were not found in any matrix habitat. Richness and abundance across breeding guilds were higher in the rainy season but frog distributions were similar across the four distances in the two seasons. Across matrix types, leaf-litter species primarily used Eucalyptus plantations, whereas water-body species primarily used coffee plantations. Bromeliad breeders were not found inside any matrix habitat. Our study highlights the importance of primary forest for bromeliad and leaf-litter breeders. We propose that water-body breeders use edge and matrix habitats to reach breeding habitats along the valleys. Including life-history characteristics, such as breeding guild, can improve predictions of frog distributions in response to edge effect and matrix types, and can guide more effective management and conservation actions.

Altered bioenergetics and developmental effects in striped marsh frog (Limnodynastes peronii) tadpoles exposed to UV treated sewage.

Effectively treating domestic wastewater so that it can be safely discharged or reused is critical for maintaining the integrity of freshwater resources, and for protecting the health of animals that rely on these systems. Amphibians are currently facing widespread population declines, so there is a particularly urgent need to investigate exposure scenarios that might result in weakened amphibian populations. Domestic sewage has received little attention as a possible factor that could influence the survival, growth and development, or general health of amphibians. However, wastewater reuse for crop irrigation and other purposes is increasing and holding ponds and constructed wetlands exist at many wastewater treatment facilities, introducing conceivable pathways that could result in the exposure of amphibians to treated wastewater. We exposed developing striped marsh frog (Limnodynastes peronii) tadpoles, to control water and 12.5, 25, 50 and 100% UV treated domestic sewage, and quantified effects on growth and development, hepatic energy reserves, and enzymatic pathways associated with detoxification and oxidative stress. Growth and development were accelerated and relative liver size was increased in exposed animals. The exposure resulted in an apparently hormetic increase in hepatic triglycerides and dose-dependent reduction in glycogen stores, as well as increased lipase and NADPH activity, indicating a general disruption to energy metabolism and/or mobilization. Contrary to expectations based on published studies with fish, we found no evidence of lipid peroxidation or induction of the detoxification enzyme Superoxide Dismutase (SOD), however, this may reflect the use of UV treatment as opposed to chlorination for disinfection. Chemical analysis and risk-based prioritization consistently identified fluoxetine, triclosan and diazinon as high-risk contaminants in the wastewater, with nonylphenol and mestranol flagged as risks during one early collection. Research is needed to explore the potential for these specific contaminants to elicit the responses identified in the present study, and to perform similar assessments using wastewater from other locations with different treatment options.

Potencial terapéutico de la sumidad de agripalma (Leonurus cardiaca L) / Potencial terapeutico da sumidade florida de agripalma (Leonurus cardiaca L) / Therapeutic potential of motherwort (Leonurus cardiaca L)

La agripalma (Leonurus cardiaca L.) es una planta herbácea, perteneciente a la familia de las Lamiáceas, originaria de Europa. En su composición destacan los terpenoides como el ajugol y la leocardina, los heterósidos fenilpropánicos como el lavandulifoliósido y los alcaloides como la estaquidrina. En Europa esta descrita desde el siglo XV la utilización de la sumidad florida de agripalma para tratar la ansiedad y alteraciones cardíacas de tipo nervioso. A nivel experimental se han descrito efectos sedantes y una acción inotrópica leve, sin embargo los estudios clínicos destinados a probar su eficacia sobre afecciones cardiovasculares son muy escasos y la mayoría antiguos. Hoy en día tiene una comercializacion muy reducida pese a tener un elevado grado de seguridad y a la posibilidad de su registro tanto en forma de medicamento tradicional a base de plantas (MTP), ya que cuenta con monografía de la Agencia Europea del Medicamento (EMA), como en la de complemento alimenticio (CA) (AU)

Motherwort (Leenurus cardiaca L.) is a herbaceous plant native to Europe, belonging to the family of Lamiaceae. lts composition include terpenoids as ajugol and leocardin, phenilpropane glycosides as lavandulifolioside and alkaloids as stachydrine. In Europe, the use of motherwort flowering tops for treating anxiety disorders and heart diseases of nervous origin is known from the fifteenth century. Experimentally, sedative activity and mild inotropic action have been described, however clinical studies to prove its effectiveness on cardiovascular disorders are scarce and mostly ancient. Nowadays, it has a small market, despite having a high degree of safety and the possibility of registration both as a traditional herbal medicinal product (TMP), since it has monograph of the European Medicines Agency (EMA), or as food supplement (CA) (AU)

A agripalma (Leonurus cardiaca L.) é uma planta herbécea, pertencente a familia das Lamiaceae, originária da Europa. Na sua composigao destacam-se os terpenóides como ajugol e leocardina, os heterósidos fenilpropánicos come lavandulifoliósido e os alcalóides como estaquidrina. Na Europa está descrita desde o seculo XV a utilizaçao da sumidade florida de agripalma para a ansiedade e alteraçoes cardíacas de tipo nervoso. A nível experimental estao descritos efeitos sedativos e açao inotropica leve, porem os estudos clínicos destinades a prevar a sua eficácia sobre as doenças cardiovasculares sao rares e a maioria antigos. Actualmente tem uma comercializaçao muito pequena, apesar de ter um alto grau de segurança e tem a possibilidade de ser registada tanto na forma de medicamento tradicional a base de plantas (MTP), por ter monografia da Agencia Eurepeia de Medicamentos (EMA), como na forma de suplemente alimentar (SA) (AU)

Sound Classification and Call Discrimination Are Decoded in Order as Revealed by Event-Related Potential Components in Frogs.

Species that use communication sounds to coordinate social and reproductive behavior must be able to distinguish vocalizations from nonvocal sounds as well as to identify individual vocalization types. In this study we sought to identify the neural localization of the processes involved and the temporal order in which they occur in an anuran species, the music frog Babina daunchina. To do this we measured telencephalic and mesencephalic event-related potentials (ERPs) elicited by synthesized white noise (WN), highly sexually attractive (HSA) calls produced by males from inside nests and male calls of low sexual attractiveness (LSA) produced outside of nests. Each stimulus possessed similar temporal structures. The results showed the following: (1) the amplitudes of the first negative ERP component (N1) at ∼ 100 ms differed significantly between WN and conspecific calls but not between HSA and LSA calls, indicating that discrimination between conspecific calls and nonvocal sounds occurs in ∼ 100 ms, (2) the amplitudes of the second positive ERP component (P2) at ∼ 200 ms in the difference waves between HSA calls and WN were significantly higher than between LSA calls and WN in the right telencephalon, implying that call characteristic identification occurs in ∼ 200 ms and (3) WN evoked a larger third positive ERP component (P3) at ∼ 300 ms than conspecific calls, suggesting the frogs had classified the conspecific calls into one category and perceived WN as novel. Thus, both the detection of sounds and the identification of call characteristics are accomplished quickly in a specific temporal order, as reflected by ERP components. In addition, the most dynamic ERP patterns appeared in the left mesencephalon and the right telencephalon, indicating the two brain regions might play key roles in anuran vocal communication.

Progesterone and prostaglandin F2α induce species-typical female preferences for male sexual displays in Cope's gray treefrog (Hyla chrysoscelis).

Endocrine systems play critical roles in facilitating sexual behavior in seasonally breeding vertebrates. Much of the research exploring this topic has focused on the endocrine correlates of signaling behavior in males and sexual proceptivity in females. What is less understood is how hormones promote the expression of the often complex and highly selective set of stimulus-response behaviors that are observed in naturally breeding animals. In female frogs, phonotaxis is a robust and sensitive bioassay of mate choice and is exhibited by gravid females during the breeding season. In stark contrast, females exhibit low phonotactic responsiveness outside the breeding season, but the administration of hormones can induce sexual proceptivity. Here we test the hypothesis that manipulation of a minimal set of reproductive hormones-progesterone and prostaglandin F2α-are capable of evoking not only proceptive behavior in non-breeding females, but also the patterns of intraspecific selectivity for male sexual displays observed in gravid females tested during the breeding season. Specifically, we investigated whether preferences for faster call rates, longer call durations, and higher call efforts were similar between breeding and hormone-treated females of Cope's gray treefrog (Hyla chrysoscelis). Hormone injections induced patterns of selective phonotaxis in non-breeding females that were remarkably similar to those observed in breeding females. These results suggest that there may be an important contribution of hormonal pleiotropy in regulating this complex, acoustically-guided sexual behavior. Our findings also support the idea that hormonal induction could be used to evaluate hypotheses about selective mate choice, and its underlying mechanisms, using non-breeding females.

Meeting ultraviolet B radiation requirements of amphibians in captivity: a case study with mountain chicken frogs (Leptodactylus fallax) and general recommendations for pre-release health screening.

Conservation breeding programmes are a tool used to prevent amphibian extinctions. The husbandry requirements of amphibians are complex. Ongoing research is needed to ensure optimal management of those captive-bred animals destined, in particular, for reintroduction. The UV-B and vitamin D3 requirements of amphibians are largely unknown. Metabolic bone disease has been reported in a number of species. These include the Critically Endangered mountain chicken frog (Leptodactylus fallax) reared in captivity on diets supplemented with a high-calcium multivitamin and mineral supplement containing vitamin D3 but without UV-B provision. Captive-bred L. fallax being reared for reintroduction to Montserrat were provided with UV-B radiation from metamorphosis and were fed on insects supplemented with vitamins and minerals. Overlapping heat, light and UV-B gradients were provided, mimicking what we believe best represents the natural situation and thereby facilitated self-regulation of UV-B exposure. A subset of 10 frogs was periodically radiographed to assess skeletal health. Radiographic bone density and anatomical integrity appeared unremarkable when compared with a wild caught L. fallax. In addition to other routine health-screening, we recommend that radiography be performed to a structured schedule on a subset of all captive-bred and reared amphibians to assess skeletal health and to gauge the appropriateness of captive husbandry. We demonstrate here that, through the appropriate provision of a combination of both UV-B radiation and dietary supplementation, L. fallax can be bred and reared in captivity with healthy skeletal development.

Treefrogs as animal models for research on auditory scene analysis and the cocktail party problem.

The perceptual analysis of acoustic scenes involves binding together sounds from the same source and separating them from other sounds in the environment. In large social groups, listeners experience increased difficulty performing these tasks due to high noise levels and interference from the concurrent signals of multiple individuals. While a substantial body of literature on these issues pertains to human hearing and speech communication, few studies have investigated how nonhuman animals may be evolutionarily adapted to solve biologically analogous communication problems. Here, I review recent and ongoing work aimed at testing hypotheses about perceptual mechanisms that enable treefrogs in the genus Hyla to communicate vocally in noisy, multi-source social environments. After briefly introducing the genus and the methods used to study hearing in frogs, I outline several functional constraints on communication posed by the acoustic environment of breeding "choruses". Then, I review studies of sound source perception aimed at uncovering how treefrog listeners may be adapted to cope with these constraints. Specifically, this review covers research on the acoustic cues used in sequential and simultaneous auditory grouping, spatial release from masking, and dip listening. Throughout the paper, I attempt to illustrate how broad-scale, comparative studies of carefully considered animal models may ultimately reveal an evolutionary diversity of underlying mechanisms for solving cocktail-party-like problems in communication.

Impacts of UVB provision and dietary calcium content on serum vitamin D3 , growth rates, skeletal structure and coloration in captive oriental fire-bellied toads (Bombina orientalis).

Many amphibian species are dependent on ex situ conservation interventions for their long-term persistence. However, projects have been jeopardised by husbandry issues involving poor calcium metabolism and nutritional metabolic bone disease (NMBD). Healthy calcium metabolism requires appropriate dietary calcium content and access to vitamin D3 . In many animals, vitamin D3 can be photobiosynthesised in skin exposed to UVB radiation, as well as extracted from the diet, but the extent of vitamin D3 photobiosynthesis in amphibians is poorly known. Additionally, prey insects for captive amphibians are deficient in calcium and calcium content must be artificially increased, but the effects of different levels of augmentation and their interaction with UVB exposure are also little understood. We fed captive fire-bellied toads (Bombina orientalis) with crickets augmented to contain 5% and 10% calcium and housed them with and without UVB exposure. Despite additional dietary vitamin D3 supplementation, we found that toads exposed to UVB radiation exhibited significantly higher serum vitamin D3 levels, indicating that this species may partly rely on photobiosynthesis sources of vitamin D3 . These data are the first to show a direct link between UVB exposure and serum vitamin D3 in an amphibian. We found significant positive effects of UVB exposure and 10% dietary calcium content on skeletal structure, as well as complex interactions between treatments. We also found UVB radiation exposure resulted in more rapid natural coloration acquisition. Together, this indicates that standard calcium plus vitamin D3 supplementation methods may not fully substitute for UVB exposure and for increased feeder insect calcium content. This may have implications for the success of ex situ amphibian conservation, as well as for the welfare of captive amphibians in general. Our data lend support for the provision of UVB radiation for captive, basking amphibians.