Litoria Ewingii Research Articles

Unravelling male advertisement call variability in the brown tree frog (Litoria ewingii) complex by using citizen science

In the face of global biodiversity declines, conservation measures are urgently needed. However, our lack of knowledge on species’ diversity, distributions and population trends presents a major obstacle, particularly for morphologically cryptic species. Field surveys to gather this information present a logistical challenge, but the rapid rise of citizen science presents a new opportunity, generating huge volumes of data rapidly across a species’ range. We use data from a national citizen-science project to document the distribution and advertisement call variability in a group of five morphologically conserved treefrogs. Using 542 male advertisement calls across the entire range of five species, we found considerable levels of acoustic variation both among and within species. Note shape, notes per call and positions of the longest and shortest notes were most useful to delineate species. Our research will assist in species delineation and identification in the field and form the basis for a more accurate understanding of species diversity and distributions in the brown tree frog complex. We also demonstrated the utility of citizen science in documenting species distributions and diversity.

Systematic assessment of the brown tree frog (Anura: Pelodryadidae: Litoria ewingii) reveals two endemic species in South Australia.

The brown tree frog (Litoria ewingii) is a relatively widespread, commonly encountered pelodryadid frog from south-eastern Australia, known for its characteristic whistling call. The distribution of Litoria ewingii spans over more than 350,000 km2, encompassing a range of moist temperate habitats, and is fragmented by well-known biogeographic barriers. A preliminary analysis of mitochondrial DNA sequences revealed evidence for deep phylogenetic structure between some of these fragmented populations. In this study, we sought to re-evaluate the systematics and taxonomy of Litoria ewingii sensu lato by analysing variation in nuclear and mitochondrial DNA, adult morphology and male advertisement calls throughout the species range. Our analyses reveal two additional, deeply divergent and allopatric lineages in South Australia. We herein re-describe Litoria ewingii from Tasmania, southern New South Wales, Victoria and south-eastern South Australia, resurrect the name Litoria calliscelis for a species occurring in the Mount Lofty Ranges and Fleurieu Peninsula in South Australia, and describe a new species, Litoria sibilus sp. nov., endemic to Kangaroo Island.

Bigger and bolder: Widespread agricultural pollutant 17β-trenbolone increases growth and alters behaviour in tadpoles (Litoria ewingii)

Endocrine-disrupting chemicals—compounds that directly interfere with the endocrine system of exposed animals—are insidious environmental pollutants that can disrupt hormone function, even at very low concentrations. The dramatic impacts that some endocrine-disrupting chemicals can have on the reproductive development of wildlife are well documented. However, the potential of endocrine-disrupting chemicals to disrupt animal behaviour has received far less attention, despite the important links between behavioural processes and population-level fitness. Accordingly, we investigated the impacts of 14 and 21-day exposure to two environmentally realistic levels of 17β-trenbolone (4.6 and 11.2 ng/L), a potent endocrine-disrupting steroid and agricultural pollutant, on growth and behaviour in tadpoles of an anuran amphibian, the southern brown tree frog (Litoria ewingii). We found that 17β-trenbolone altered morphology, baseline activity and responses to a predatory threat, but did not affect anxiety-like behaviours in a scototaxis assay. Specifically, we found that tadpoles exposed to our high-17β-trenbolone treatment were significantly longer and heavier at 14 and 21 days. We also found that tadpoles exposed to 17β-trenbolone showed higher levels of baseline activity, and significantly reduced their activity following a simulated predator strike. These results provide insights into the wider repercussions of agricultural pollutants on key developmental and behavioural traits in aquatic species, and demonstrate the importance of behavioural studies in the ecotoxicological field.

Interspecific amplexus between a male Yang’s Frilled Treefrog, Kurixalus yangi Yu, Hui, Rao, and Yang 2018, and a female Brauer’s Brown Treefrog, Polypedates braueri (Vogt 1911), with recommendations for English common names

Interspecific amplexus between a male Yang’s Frilled Treefrog, Kurixalus yangi Yu, Hui, Rao, and Yang 2018, and a female Brauer’s Brown Treefrog, Polypedates braueri (Vogt 1911), with recommendations for English common names

Traffic noise drives an immediate increase in call pitch in an urban frog

AbstractNoise pollution is an underappreciated component of global environmental change and can impact species that have a strong reliance on acoustic communication. In urban areas, traffic noise can interfere with the ability of animals to communicate and complete essential aspects of their daily lives. We investigated the impact of traffic noise on the calling behaviour of the brown tree frog (Litoria ewingii) in south‐eastern Melbourne, the fastest‐growing human population centre in Australia. We placed six acoustic recorders at increasing distances from a busy suburban road and recorded the calling behaviour (call pitch and call rate) of brown tree frogs immediately before and after loud traffic noises, and in response to different chorus sizes. Traffic noise resulted in a significant, but short‐term, increase in call pitch in the brown tree frog. Both call pitch and call rate decreased with increasing distance from the road, yet traffic noise still resulted in increased call pitch even 200–300 m from the road. Conversely, although traffic noise increased call pitch across all chorus sizes of the brown tree frog, larger chorus sizes were associated with decreased call pitch. Our study highlights the pervasive, and sustained, impact that anthropogenic noise can have on urban frog populations.

A new species of frog in the Litoria ewingii species group (Anura: Pelodryadidae) from south-eastern Australia.

Population declines and range contractions among Australian frogs that commenced in the early 1980s continue in some species that were once widespread. The generality of this pattern has been difficult to discern, especially for those species that are encountered rarely because they have restricted periods of calling activity with poorly defined habitat preferences, and are not common. Several lines of evidence indicate that Litoria littlejohni is such a species. This frog was once known from mid-eastern New South Wales to eastern Victoria, and evidence from wildlife atlas databases and targeted searches indicate that it has declined in large portions of its former range, leaving several populations that are isolated, in some cases restricted in distribution, and of small size. We investigated the relationships among populations using mitochondrial ND4 nucleotide sequences and single nucleotide polymorphisms (SNPs) from the nuclear genome. We found that northern and southern populations form two highly divergent genetic groups whose distributions abut at the southern margin of the Sydney Basin Bioregion and these genetic groups also show divergence in morphology and male advertisement calls. Here we describe the populations to the south of the Sydney Basin Bioregion as a new species and provide information on its distribution and ecology. In light of the apparent isolation and small size of known populations of the new species and the consequent restriction of the range of L. littlejohni, we assessed the conservation status of both species.

Native amphibian larvae exhibit higher upper thermal limits but lower performance than their introduced predator Gambusia affinis

Information on the thermal limits and physiology of ectothermic amphibians is crucial to our understanding of their ecology in the natural environment, particularly with predicted global changes in climate. We documented the thermal limits of larvae of three amphibian species native to Hong Kong, and their introduced, invasive predator, the mosquitofish (Gambusia affinis). We then used larvae of the brown tree frog Polypedates megacephalus as a model amphibian to further investigate growth, oxygen consumption rate and heat shock protein expression with changes in thermal regime. We found that G. affinis was the most tolerant of low temperatures but also the least tolerant of high temperatures. Despite the higher thermal tolerance of the amphibian larvae, further investigation on P. megacephalus demonstrated that optimal temperatures for physiological performance fall within a range of 18.0–21.6 °C, which is far lower than its upper thermal limit, implying that thermal stress occurs during part of the larval stage under natural environmental conditions. This could mean a reduction in their capacity to deal with other stressors such as pollution and predators, and that G. affinis may have an advantage over native amphibians.

Functional analysis for gut microbes of the brown tree frog (Polypedates megacephalus) in artificial hibernation.

BackgroundAnnual hibernation is an adaptation that helps many animals conserve energy during food shortage in winter. This natural cycle is also accompanied by a remodeling of the intestinal immune system, which is an aspect of host biology that is both influenced by, and can itself influence, the microbiota. In amphibians, the bacteria in the intestinal tract show a drop in bacterial counts. The proportion of pathogenic bacteria is greater in hibernating frogs than that found in nonhibernating frogs. This suggests that some intestinal gut microbes in amphibians can be maintained and may contribute to the functions in this closed ecosystem during hibernation. However, these results were derived from culture-based approaches that only covered a small portion of bacteria in the intestinal tract.MethodsIn this study, we use a more comprehensive analysis, including bacterial appearance and functional prediction, to reveal the global changes in gut microbiota during artificial hibernation via high-throughput sequencing technology.ResultsOur results suggest that artificial hibernation in the brown tree frog (Polypedates megacephalus) could reduce microbial diversity, and artificially hibernating frogs tend to harbor core operational taxonomic units that are rarely distributed among nonhibernating frogs. In addition, artificial hibernation increased significantly the relative abundance of the red-leg syndrome-related pathogenic genus Citrobacter. Furthermore, functional predictions via PICRUSt and Tax4Fun suggested that artificial hibernation has effects on metabolism, disease, signal transduction, bacterial infection, and primary immunodeficiency.ConclusionsWe infer that artificial hibernation may impose potential effects on primary immunodeficiency and increase the risk of bacterial infections in the brown tree frog.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3318-6) contains supplementary material, which is available to authorized users.

Short- and long-term consequences of developmental saline stress: impacts on anuran respiration and behaviour.

Secondary salinization has been identified as a major stressor to amphibians. Exposure to elevated salinity necessitates physiological adjustments and biochemical changes that may be energetically demanding. As such, exposure to non-lethal levels of salinity during development could potentially alter anuran metabolic rates and individual performance in both pre- and post-metamorphic life stages. We investigated the effects of non-lethal levels of salinity on metamorphic traits (time to reach metamorphosis and metamorphic mass), tadpole oxygen consumption, escape response behaviour (pre- and post-metamorphosis) and foraging ability post-metamorphosis in two native Australian frog species, the southern brown tree frog (Litoria ewingii) and the striped marsh frog (Limnodynastes peronii). We found that both Lit. ewingii and Lim. peronii exhibited differences in metamorphic traits in response to elevated salinity. Neither species showed significant change in oxygen consumption during development in response to salinity, relative to freshwater controls. Both species displayed impaired escape response behaviours in response to salinity during larval development, but flow-on effects to adult escape response behaviours and foraging performance were species-specific. Our results show that the influence of stressors during development can have consequences for anuran physiology and behaviour at multiple life stages, and emphasize the need for studies that examine the energetics of anuran responses in order to better understand the responses of biota to stressful environments.

Interspecific variation in the phenology of advertisement calling in a temperate Australian frog community.

Spatial and temporal partitioning of resources underlies the coexistence of species with similar niches. In communities of frogs and toads, the phenology of advertisement calling provides insights into temporal partitioning of reproductive effort and its implications for community dynamics. This study assessed the phenology of advertisement calling in an anuran community from Melbourne, in southern Australia. We collated data from 1432 surveys of 253 sites and used logistic regression to quantify seasonality in the nightly probability of calling and the influence of meteorological variables on this probability for six species of frogs. We found limited overlap in the predicted seasonal peaks of calling among these species. Those shown to have overlapping calling peaks are unlikely to be in direct competition, due to differences in larval ecology (Crinia signifera and Litoria ewingii) or differences in calling behavior and acoustics (Limnodynastes dumerilii and Litoria raniformis). In contrast, closely related and ecologically similar species (Crinia signfera and Crinia parinsignifera;Litoria ewingii and Litoria verreauxii) appear to have staggered seasonal peaks of calling. In combination with interspecific variation in the meteorological correlates of calling, these results may be indicative of temporal partitioning of reproductive activity to facilitate coexistence, as has been reported for tropical and temperate anurans from other parts of the globe.

Temperature-dependent acute toxicity of methomyl pesticide on larvae of 3 Asian amphibian species.

Relative to other animal taxa, ecotoxicological studies on amphibians are scarce, even though amphibians are experiencing global declines and pollution has been identified as an important threat. Agricultural lands provide important habitats for many amphibians, but often these lands are contaminated with pesticides. The authors determined the acute toxicity, in terms of 96-h median lethal concentrations, of the carbamate pesticide methomyl on larvae of 3 Asian amphibian species, the Asian common toad (Duttaphrynus melanostictus), the brown tree frog (Polypedates megacephalus), and the marbled pygmy frog (Microhyla pulchra), at 5 different temperatures (15 °C, 20 °C, 25 °C, 30 °C, and 35 °C) to examine the relationships between temperature and toxicity. Significant interspecific variation in methomyl sensitivity and 2 distinct patterns of temperature-dependent toxicity were found. Because high proportions of malformation among the surviving tadpoles were observed, a further test was carried out on the tree frog to determine effect concentrations using malformation as the endpoint. Concentrations as low as 1.4% of the corresponding 96-h median lethal concentrations at 25 °C were sufficient to cause malformation in 50% of the test population. As the toxicity of pesticides may be significantly amplified at higher temperatures, temperature effects should not be overlooked in ecotoxicological studies and derivation of safety limits in environmental risk assessment and management.

Anuran developmental plasticity loss: the cost of constant salinity stress

In animals with a complex life cycle, changes in biotic and abiotic conditions during development can alter growth and maturation rates, causing carry-over effects in postmetamorphic phenotypes. In anurans, this developmental plasticity can result in a trade-off between length of larval period and body size at metamorphosis in stressful environments. Secondary salinisation has been identified as a substantial stressor to amphibians; however, little is known about how salinity-induced developmental plasticity differs between anuran populations. We examined differences in survival, time to metamorphosis, size at metamorphosis (mass and snout–vent length) and body condition at metamorphosis in response to elevated salinity in three populations of the brown tree frog (Litoria ewingii). Significant differences in size at metamorphosis between salinity treatments were observed in tadpoles sourced from freshwater wetlands and ephemeral wetlands, with tadpoles showing a reduced mass and snout–vent length at metamorphosis in the higher-salinity treatment. There were no significant differences in metamorphic traits between salinity treatments in tadpoles sourced from a consistently brackish wetland, suggesting either an erosion of developmental plasticity in response to elevated salinity, or the magnitude of salinity required to alter developmental traits is higher in this population. Our results indicate that environmental conditions of source populations need to be considered when studying life-history adaptations in response to environmental change.

Anesthesia of tadpoles of the southern brown tree frog (Litoria ewingii) with isoeugenol (Aqui-S).

Because anesthesia of amphibian larvae is currently problematic, the aim of this study was to test if a fish anesthetic containing approximately 500 g/L isoeugenol (Aqui-S) could anesthetize tadpoles. Three concentrations of Aqui-S were tested (of commercial solution 10, 20, and 50 microl/L) with 10 tadpoles of the southern brown tree frog (Litoria ewingii) per concentration. Tadpoles were anesthetized for 15 min in water at 18-20 degrees C. Anesthetic criteria included four time points and two heart rates: time to loss of response to stimulation, loss of righting reflex, and return of spontaneous movement and of righting reflex after removal from Aqui-S, and heart rate after induction and after 15 min of anesthesia. No tadpole showed signs of irritation or excitement, no tadpole died, and all returned to normal behavior. The times for induction of anesthesia (time to loss of righting reflex) decreased significantly (P < 0.001) with increasing concentration: means of 27.5, 16.8, and 5.7 min for 10, 20, and 50 microl/L, respectively. Heart rate after 15 min of anesthesia was significantly lower (P = 0.006) for concentrations of 20 and 50 microl/L compared to 10 microl/L. At the lowest concentration (10 microl/L), anesthesia was light. Recovery times were significantly delayed (P < 0.001) for 50 microl/L, approximately double those for the lower concentrations. Spontaneous movements began at 56.9, 51.4, and 119 min for 10, 20, and 50 microl/L Aqui-S, respectively. Although Aqui-S did anesthetize tadpoles at all three concentrations, with 20 microl/L being the most suitable, additional longer-term toxicity testing is required prior to its adoption as an anesthetic agent for this species. Anesthesia, isoeugenol, Litoria ewingii, southern brown tree frog, tadpoles.

The hormonal regulation of color changes in the sexually dichromatic frog Buergeria robusta.

During the breeding season, dynamic changes in body coloration are regularly observed in the male brown tree frog Buergeria robusta. This study investigated the hypothesis that this sexual dichromatism in male B. robusta is mediated through hormonal regulation. Frogs were exogenously injected with testosterone (T) or estradiol (E2). This manipulation revealed that the body coloration (hue, brightness, and saturation) of the male frog increased significantly (i.e., the brilliant yellow color developed) in response to T but not in response to E2. Concurrently, the levels of expression of brain-derived neurotrophic factor (BDNF) and pituitary adenylate cyclase-activating polypeptide (PACAP) in the pituitary gland were reduced in frogs whose coloration was pale brown on a yellow background. In particular, the weakest expressions of BDNF, PACAP, and PACAP type II receptors (VPAC-1R) were found in male frogs with a brilliant yellow body color during the breeding season regardless of background color. These changes may decrease α-melanocyte-stimulating hormone production associated with the PACAP receptors (VPAC-1R), resulting in the aggregation of black pigment in melanophores and the production of a brilliant yellow body color. The effects of hormones on skin coloration were further examined in isolated skin in vitro. The results of this investigation showed that the dispersion of xanthophores was stimulated by T or prolactin (PRL) and that the melanophores were aggregated by melatonin (MEL) but not by E2. Furthermore, yellow pigments in the xanthophores were significantly dispersed following the PRL+T treatment. In the T+MEL, PRL+MEL, and T+PRL+MEL treatments, xanthophores were dispersed, and melanophores were aggregated and subsequently moved to the low spongiosum layer of the dorsal skin, causing the increase in yellow coloration. These results reveal that multiple hormones play major roles in the regulation of the brilliant yellow coloration of male B. robusta by high plasma T during the breeding season.

Observations on the Calling Behaviour of the Jervis Bay Tree FrogLitoria jervisiensis

In coastal New South Wales most frog species are observed calling and breeding in the warmer months of the year. One exception to this is the Jervis Bay Tree Frog Litoria jervisiensis, which displays a preference for calling and breeding in the cooler months of the year. There are some discrepancies in the published literature on this species, in part due to past taxonomic confusion and some present difficulties in identifying members of the Litoria ewingi species-group. At one site on the Kurnell Peninsula near the geographic centre of this species range, surveys conducted from 2008-2009 revealed that the core period for calling for L. jervisiensis was April to July. Opportunistic observations of territorial behaviour and dynamic sexual dichromatism were also recorded.

Skin Gland Morphology and Secretory Peptides in NaturalizedLitoriaSpecies in New Zealand

Abstract The defensive secretions of anurans contain a large array of chemical compounds that are synthesized in the granular glands and released onto the skin. We used histological and peptidomic analyses to investigate the skin glands and their products in Litoria aurea, Litoria ewingii, and Litoria raniformis, which were introduced to New Zealand from Australia approximately 150 yr ago. The skin glands were induced to release their product by either norepinephrine or electric stimulation. Granular glands in all three species are distributed evenly in both dorsal and ventral skin and share morphological features common in other anurans, such as a contractile myoepithelium that surrounds the syncytial secretory unit. However, differences are observed in the granular ultrastructure between L. ewingii and the more closely related L. aurea and L. raniformis. The latter have larger glands with granules that are opaque and contain homogeneously spaced, diaphanous vesicles, whereas the substructure of the gran...

Shading Mediates the Interaction Between an Amphibian and a Predatory Fly

Abiotic factors have been demonstrated to mediate predator–prey interactions in aquatic and terrestrial communities. For amphibians changes in pH, pollutants, temperature, ultraviolet-B, and moisture levels can alter behavioral, developmental, and physiological aspects of predator–prey relationships resulting in reduced survivorship. Some rhacophorid frogs deposit eggs as foam nests on vegetation overhanging water, presumably as an adaptive response to the risk of egg predation by aquatic animals. Terrestrial insects are known to feed on frog embryos in foam nests, and anecdotal observations in tropical East Asia and Southeast Asia suggested that predation of frog embryos by dipteran fly larvae may be higher in unshaded areas. To understand the role that shading may play in this relationship, I compared survival of embryos of the Brown Tree Frog (Polypedates megacephalus) infested by larvae of a calliphorid fly (Phumosia coomani) in shaded and unshaded conditions in a controlled field study. When foam nests were infested by flies, survival to hatching was more than 15 times higher in shaded than in unshaded nests. Less than 1% of frog embryos survived to hatching in unshaded, infested foam nests which contained twice as many fly larvae as shaded nests. These results combine to suggest that both colonization and predation by flies are higher in unshaded foam nests. The mechanism triggering increased colonization of unshaded nests is unclear, but warmer temperatures on the surfaces of unshaded foam nests may decrease hatching time for fly eggs, permitting fly larvae to move inside the foam nest more quickly and begin feeding on amphibian embryos. Although the risk of predation by calliphorid flies is higher for P. megacephalus in unshaded areas, the benefits of developing in warmer conditions, including faster embryonic and larval development and larger size at metamorphosis, may offset this risk.

Anthropogenic noise constrains acoustic communication in urban-dwelling frogs

Urban noise may hinder acoustic communication in a diversity of animal groups by reducing the distance over which vocal signals can be detected. Given the importance of such signals for mate attraction and territory defence, this acoustic interference may have wide-ranging consequences for individual fitness. I will present a mathematical model of the active space of frog calls in urban noise as a function of body size. Despite having lower auditory thresholds, larger species with lower-frequency calls are predicted to suffer the greatest reduction in communication distance in noisy urban environments. During a field study in Melbourne, Australia, my colleagues and I found that the southern brown tree frog Litoria ewingii called at a higher frequency in traffic noise. However, modeling indicates that the observed frequency shift would confer only a modest increase in active space. Furthermore, as females of certain frog species appear to prefer lower-frequency advertisement calls, this strategy may improve the audibility of calls but reduce attractiveness to potential mates. Calling more loudly would result in a larger increase in active space, but the high metabolic cost of this strategy could limit chorus tenure and ultimately reduce breeding success.

Heavy metal pollution negatively correlates with anuran species richness and distribution in south‐eastern Australia

AbstractHeavy metal pollution has likely played an important role in global biodiversity decline, but there remains a paucity of information concerning the effects of metals on amphibian diversity. This study assessed anuran species richness and distribution in relation to sediment metal content and water chemistry in wetlands located along the Merri Creek corridor in Victoria, south‐eastern Australia. Anurans were present in 60% (21/35) of study sites, with a total of six species detected: the eastern common froglet (Crinia signifera), the eastern sign‐bearing froglet (Crinia parinsignifera), the southern brown tree frog (Litoria ewingii), the growling grass frog (Litoria raniformis), the eastern banjo frog (Limnodynastes dumerilii) and the spotted marsh frog (Limnodynastes tasmaniensis). Mean species richness was 1.77 ± 0.32 per site, and species richness ranged from zero to six species per site. Across sites, species richness correlated negatively with sediment concentrations of six heavy metals: copper, nickel, lead, zinc, cadmium and mercury. Species richness also correlated negatively with wetland water electrical conductivity (a proxy for salinity) and concentrations of orthophosphate. Distributions of the three most commonly observed frog species (C. signifera, L. tasmaniensis and L. ewingii) were significantly negatively associated with the total level of metal contamination at individual sites. The study is the first to provide evidence for an association between metal contamination and anuran species richness and distribution in the southern hemisphere, adding to a small but growing body of evidence that heavy metal pollution has contributed to global amphibian decline.

Strict consistency between genetic and topographic landscapes of the brown tree frog (Buergeria robusta) in Taiwan

Taiwan presents an excellent opportunity to build a phylogeographic paradigm for fine-scaled differentiation occurring within short distances on an single island. Due to the limitation of habitat availability on the island, demographic histories of species in Taiwan were strongly influenced by glacial–interglacial cycles. Nevertheless, there are relatively few studies demonstrating such phylogeographic patterns for islands, especially in subtropical Asia. In this study, we aim to construct the genetic landscape of a philopatric stream frog Buergeria robusta by an intense and fine-scaled collection throughout the island. The deduced genetic landscape of B. robusta presented extremely high congruence with the actual topography of Taiwan. Two major lineages were found on the eastern and the western sides of Taiwan with a non-overlapping distribution, indicating the importance of the Central Mountain Range as the major biogeographic barrier. Both clades showed a strong and congruent tendency of demographic or distributional expansion in recent history based on different analyses. Population expansion of such a subtropical lowland species might be a result from a release of available habitat in post-glacial periods.