Skin Bacterial and Fungal Microbiome Responses to Diet Supplementation and Rewilding in the Critically Endangered Southern Corroboree Frog.

The composition and dynamics of the skin bacterial and fungal microbiome is thought to influence host-pathogen defence. This microbial community is shaped by host captivity, diet, and microbial interactions between bacterial and fungal components. However, there remains little understanding of how specific micronutrients influence bacterial and fungal microbiome composition and their inter-domain interactions during rewilding of captive-bred animals.

Dietary carotenoid supplementation has long-term and community-wide effects on the amphibian skin microbiome.

The amphibian skin microbiome plays a crucial role in host immunity and pathogen defence, yet we know little about the environmental drivers of skin microbial variation across host individuals. Inter-individual variation in the availability of micro-nutrients such as dietary carotenoids, which are involved in amphibian immunity, may be one factor that influences skin microbial assembly across different life history stages. We compared the effect of four carotenoid supplementation regimes during different life stages on the adult skin microbiome using a captive population of the critically endangered southern corroboree frog, Pseudophryne corroboree.

Cryopreservation Cooling Rate Impacts Post-Thaw Sperm Motility and Survival in .

The cryopreservation and storage of gametes (biobanking) can provide a long-term, low-cost option for the preservation of population genetic diversity and is particularly impactful when applied to manage selective breeding within conservation breeding programs (CBPs). This study aimed to develop a sperm cryopreservation protocol for the critically endangered Booroolong frog () to capture founder genetics within the recently established (est. 2019) CBP for this species.

Application of Reproductive Technologies to the Critically Endangered Baw Baw Frog, .

Reproductive technologies (RTs) can assist integrated conservation breeding programs to attain propagation targets and manage genetic diversity more effectively. While the application of RTs to enhance the conservation management of threatened amphibians has lagged behind that of other taxonomic groups, a recent surge in research is narrowing the divide. The present study reports on the first application of RTs (hormone-induced spawning, hormone-induced sperm-release, and sperm cryopreservation) to the critically endangered Baw Baw frog, .

The Increasing Role of Short-Term Sperm Storage and Cryopreservation in Conserving Threatened Amphibian Species.

Multidisciplinary approaches to conserve threatened species are required to curb biodiversity loss. Globally, amphibians are facing the most severe declines of any vertebrate class. In response, conservation breeding programs have been established in a growing number of amphibian species as a safeguard against further extinction.

A test for plasticity in sperm motility activation in response to osmotic environment in an anuran amphibian.

Evolutionary theory predicts that selection will favor phenotypic plasticity in sperm traits that maximize fertilization success in dynamic fertilization environments. In species with external fertilization, osmolality of the fertilization medium is known to play a critical role in activating sperm motility, but evidence for osmotic-induced sperm plasticity is limited to euryhaline fish and marine invertebrates. Whether this capacity extends to freshwater taxa remains unknown.

Does dietary β-carotene influence ontogenetic colour change in the southern corroboree frog?

Ontogenetic colour change occurs in a diversity of vertebrate taxa and may be closely linked to dietary changes throughout development. In various species, red, orange and yellow colouration can be enhanced by the consumption of carotenoids. However, a paucity of long-term dietary manipulation studies means that little is known of the role of individual carotenoid compounds in ontogenetic colour change.

Amphibian reproductive technologies: approaches and welfare considerations.

Captive breeding and reintroduction programs have been established for several threatened amphibian species globally, but with varied success. This reflects our relatively poor understanding of the hormonal control of amphibian reproduction and the stimuli required to initiate and complete reproductive events. While the amphibian hypothalamo-pituitary-gonadal (HPG) axis shares fundamental similarities with both teleosts and tetrapods, there are more species differences than previously assumed.

Hormone-induced ovulation and artificial fertilisation in four terrestrial-breeding anurans.

The development and application of reproductive technologies has great potential to enhance the conservation management of threatened amphibians globally. The present study quantified the efficacy of protocols previously developed for Pseudophryne guentheri for hormonally inducing egg release and artificial fertilisation in three additional terrestrial-breeding species of Australian ground frog; namely Pseudophryne bibronii , Pseudophryne coriacea and Heleioporus eyrei . Females of each species were administered a priming dose of 0.

Evidence that genetic compatibility underpins female mate choice in a monandrous amphibian.

Mate choice for genetic benefits remains controversial, largely because few studies have estimated the relative contributions of additive and non-additive sources of genetic variation to offspring fitness. Moreover, there remains a deficit of these estimates for species where female-mate preferences have been quantified in the wild, especially species characterized by monandry or monogamy. Here, we use artificial fertilization techniques combined with a cross-classified breeding design to simultaneously test for "good genes" and "compatible genes" benefits of mate choice in the monandrous red backed toadlet (Pseudophryne coriacea).

The effect of injection and topical application of hCG and GnRH agonist to induce sperm-release in the roseate frog, .

Reproductive technologies may assist amphibian conservation breeding programs (CBPs) to achieve propagation targets and genetic management goals. However, a trial-and-error approach to protocol refinement has led to few amphibian CBPs routinely employing reproductive technologies with predictable outcomes. Additionally, while injections can be safely administered to amphibians, perceived animal welfare risks, such as injury and disease transmission, warrant the development of alternative hormone administration protocols.

Genetic benefits of extreme sequential polyandry in a terrestrial-breeding frog.

Sequential polyandry may evolve as an insurance mechanism to reduce the risk of females choosing mates who are genetically inferior (intrinsic male quality hypothesis) or genetically incompatible (genetic incompatibility hypothesis). The prevalence of such indirect benefits remains controversial, however, because studies estimating the contributions of additive and nonadditive sources of genetic variation to offspring fitness have been limited to a small number of taxonomic groups. Here, we used artificial fertilization techniques combined with a crossclassified breeding design (North Carolina Type II) to simultaneously test the "good genes hypothesis" and the "genetic incompatibility hypothesis" in the brown toadlet (Pseudophryne bibronii); a terrestrial-breeding species with extreme sequential polyandry.

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.

Sperm collection and storage for the sustainable management of amphibian biodiversity.

Current rates of biodiversity loss pose an unprecedented challenge to the conservation community, particularly with amphibians and freshwater fish as the most threatened vertebrates. An increasing number of environmental challenges, including habitat loss, pathogens, and global warming, demand a global response toward the sustainable management of ecosystems and their biodiversity. Conservation Breeding Programs (CBPs) are needed for the sustainable management of amphibian species threatened with extinction.

Effect of carotenoid class and dose on the larval growth and development of the critically endangered southern corroboree frog.

Dietary carotenoids are expected to improve vertebrate growth and development, though evidence for beneficial effects remains limited. One reason for this might be that few studies have directly compared the effects of carotenoids from different classes (carotenes versus xanthophylls) at more than one dose. Here, we tested the effect of two doses of dietary β-carotene and lutein (representing two different carotenoid classes) on the growth and development of larval southern corroboree frogs ().

Hormone-induced sperm-release in the critically endangered Booroolong frog (): effects of gonadotropin-releasing hormone and human chorionic gonadotropin.

Research into the development of reproductive technologies for amphibians has increased in recent years due to the rapid decline of amphibian species globally. Reproductive technologies have great potential to overcome captive breeding failure and improve the propagation and genetic management of threatened species. However, the incorporation of these technologies into conservation breeding programs has been protracted, primarily as a result of trial-and-error approaches to the refinement of hormone therapies.

The Role of Reproductive Technologies in Amphibian Conservation Breeding Programs.

Anthropogenic environmental change has led to unprecedented rates of species extinction, presenting a major threat to global biodiversity. Among vertebrates, amphibians have been most severely impacted, with an estimated 41% of species now threatened with extinction. In response to this biodiversity crisis, a moral and ethical obligation exists to implement proactive interventionist conservation actions to assist species recovery and decelerate declines.

Dose and life stage-dependent effects of dietary beta-carotene supplementation on the growth and development of the Booroolong frog.

Carotenoids are known for their antioxidant capacity and are considered to play an important role in vertebrate growth and development. However, evidence for their beneficial effects remains limited, possibly because very few studies have tested for dose effects across different life stages. The present study investigated the effect of various doses of dietary beta-carotene supplements on the growth and development of larval and post-metamorphic Booroolong frogs ().

The unexpected genetic mating system of the red-backed toadlet (Pseudophryne coriacea): A species with prolonged terrestrial breeding and cryptic reproductive behaviour.

Molecular technologies have revolutionized our classification of animal mating systems, yet we still know very little about the genetic mating systems of many vertebrate groups. It is widely believed that anuran amphibians have the highest reproductive diversity of all vertebrates, yet genetic mating systems have been studied in <1% of all described species. Here, we use single nucleotide polymorphisms to quantify the genetic mating system of the terrestrial breeding red-backed toadlet Pseudophryne coriacea.

Hormone-induced spawning of the critically endangered northern corroboree frog Pseudophryne pengilleyi.

Fundamental knowledge of the optimal hormone concentrations required to stimulate amplexus and spawning in breeding pairs of amphibians is currently lacking, hindering our understanding of the proximate mechanisms underpinning mating behaviour. The present study investigated the effects of: (1) the dose of a gonadotropin-releasing hormone analogue (GnRH-a) administered; (2) male-female hormone administration interval; and (3) topical application of GnRH-a, on spawning success in the northern corroboree frog. Administration of GnRH-a at doses of 0.

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.

Sperm motility activation in the critically endangered booroolong frog: the effect of medium osmolality and phosphodiesterase inhibitors.

Effective activation of sperm motility is fundamental to successful artificial fertilisation; however, studies investigating optimal procedures in amphibians are lacking. This study found the optimal osmolality of activation media for sperm motility activation and evaluated the effect of phosphodiesterase (PDE) inhibitors on sperm activation and longevity in the critically endangered booroolong frog, Litoria booroolongensis. To assess the effect of medium osmolality (10, 25, 50, 75, 100 and 200mOsmolkg) and PDE inhibitors (control, 2.

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 effect of gentamicin on sperm motility and bacterial abundance during chilled sperm storage in the Booroolong frog.

Antibiotics can inhibit bacterial contamination and extend sperm longevity during storage; a primary goal of captive facilities conducting biobanking and artificial fertilisation (AF). This study evaluated the effects of gentamicin on the short-term storage of Booroolong frog sperm. Sperm suspensions were obtained via either testis maceration, or as spermic urine, following hormonal induction of sperm-release.

Dietary Carotenoid Supplementation Enhances the Cutaneous Bacterial Communities of the Critically Endangered Southern Corroboree Frog (Pseudophryne corroboree).

The rapid spread of infectious disease has resulted in the decline of animal populations globally. Amphibians support a diversity of microbial symbionts on their skin surface that help to inhibit pathogen colonisation and reduce disease susceptibility and virulence. These cutaneous microbial communities represent an important component of amphibian immune defence, however, very little is known about the environmental factors that influence the cutaneous microbiome.