Broad-scale pesticide screening finds anticoagulant rodenticide and legacy pesticides in Australian frogs.

Pesticide contamination poses a significant threat to non-target wildlife, including amphibians, many of which are already highly threatened. This study assessed the extent of pesticide exposure in dead frogs collected during a mass mortality event across eastern New South Wales, Australia between July 2021 and March 2022. Liver tissue from 77 individual frogs of six species were analysed for >600 legacy and contemporary pesticides, including rodenticides. More than a third (36 %) of the liver samples contained at least one of the following pesticides: brodifacoum, dieldrin, DDE, heptachlor/heptachlor epoxide, fipronil sulfone, and 2-methyl-4-chlorophenoxyacetic acid (MCPA). Brodifacoum, a second-generation anticoagulant rodenticide, was found in four of the six frog species analysed: the eastern banjo frog (Limnodynastes dumerilii), cane toad (Rhinella marina), green tree frog (Litoria caerulea) and Peron's tree frog (Litoria peronii). This is the first report of anticoagulant rodenticide detected in wild amphibians, raising concerns about potential impacts on frogs and extending the list of taxa shown to accumulate rodenticides. Dieldrin, a banned legacy pesticide, was also detected in two species: striped marsh frog (Limnodynastes peronii) and green tree frog (Litoria caerulea). The toxicological effects of these pesticides on frogs are difficult to infer due to limited comparable studies; however, due to the low frequency of detection the presence of these pesticides was not considered a major contributing factor to the mass mortality event. Additional research is needed to investigate the effects of pesticide exposure on amphibians, particularly regarding the impacts of second-generation anticoagulant rodenticides. There is also need for continued monitoring and improved conservation management strategies for the mitigation of the potential threat of pesticide exposure and accumulation in amphibian populations.

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.

Phylogeography, hybrid zones and contemporary species boundaries in the south-eastern Australian smooth frogs (Anura: Myobatrachidae: Geocrinia).

Paleo-climatic fluctuations have driven episodic changes in species distributions, providing opportunities for populations to diverge in isolation and hybridise following secondary contact. Studies of phylogeographic diversity and patterns of gene flow across hybrid zones can provide insight into contemporary species boundaries and help to inform taxonomic and conservation inferences. Here we explore geographic diversity within the acoustically divergent yet morphologically conserved south-eastern Australian smooth frog complex and assess gene flow across a narrow hybrid zone using mitochondrial nucleotide sequences and nuclear genome-wide single nucleotide polymorphisms. Our analyses reveal the presence of an evolutionarily distinct taxon restricted to the Otway Plains and Ranges, Victoria, which forms a narrow (9-30 km wide), spatiotemporally stable (>50 years) hybrid zone with Geocrinia laevis, which we describe herein as a new species.

An invasive pathogen drives directional niche contractions in amphibians.

Global change is causing an unprecedented restructuring of ecosystems, with the spread of invasive species being a key driver. While population declines of native species due to invasives are well documented, much less is known about whether new biotic interactions reshape niches of native species. Here we quantify geographic range and realized-niche contractions in Australian frog species following the introduction of amphibian chytrid fungus Batrachochytrium dendrobatidis, a pathogen responsible for catastrophic amphibian declines worldwide. We show that chytrid-impacted species experienced proportionately greater contractions in niche breadth than geographic distribution following chytrid emergence. Furthermore, niche contractions were directional, with contemporary distributions of chytrid-impacted species characterized by higher temperatures, lower diurnal temperature range, higher precipitation and lower elevations. Areas with these conditions may enable host persistence with chytrid through lower pathogenicity of the fungus and/or greater demographic resilience. Nevertheless, contraction to a narrower subset of environmental conditions could increase host vulnerability to other threatening processes and should be considered in assessments of extinction risk and during conservation planning. More broadly, our results emphasize that biotic interactions can strongly shape species realized niches and that large-scale niche contractions due to new species interactions-particularly emerging pathogens-could be widespread.

A new species of Delma Gray 1831 (Squamata: Pygopodidae) from the Hunter Valley and Liverpool Plains of New South Wales.

The Striped Legless Lizard Delma impar Fischer 1882 occurs in grasslands of south-eastern Australia where it is sometimes sympatric with several congeners. Previous molecular analyses have revealed mitochondrial divergence between the northern and southern lineages of D. impar, but the status of a recently detected population assigned to the species in the Hunter Valley of New South Wales (NSW), 250 km north of other populations, remains unknown. Via morphological and molecular analyses across the known distribution of D. impar, we distinguish the population from the Hunter Valley, Liverpool Plains region as a unique taxon and describe this species as D. vescolineata sp. nov. The new species is divergent from D. impar in both nuclear and mitochondrial analysis, and differing facial scalation and colour patterning from the limited number of specimens available. The degree of mitochondrial and nuclear divergence between the previously identified northern and southern clades of D. impar is consistent with a single species. This is the first description of a Delma from south-eastern Australia since 1974, but the second new vertebrate taxon identified from the Hunter Valley region within a decade. Our research highlights our incomplete understanding of the biodiversity of the Hunter Valley region, despite its relative proximity to populous areas. The range of D. vescolineata sp. nov. is restricted and heavily overlaps with major habitat modification including mining and grazing, with no parks or reserves in this distribution protecting the species grassland habitat. Gathering more data to understand conservation status of this species should be a priority.

Anthropogenic habitat modification alters calling phenology of frogs.

Anthropogenic habitat modification significantly challenges biodiversity. With its intensification, understanding species' capacity to adapt is critical for conservation planning. However, little is known about whether and how different species are responding, particularly among frogs. We used a continental-scale citizen science dataset of >226,000 audio recordings of 42 Australian frog species to investigate how calling-a proxy for breeding-phenology varied along an anthropogenic modification gradient. Calling started earlier and breeding seasons lengthened with increasing modification intensity. Breeding seasons averaged 22.9 ± 8.25 days (standard error) longer in the most modified compared to the least modified regions, suggesting that frog breeding activity was sensitive to habitat modification. We also examined whether calls varied along a modification gradient by analysing the temporal and spectral properties of advertisement calls from a subset of 441 audio recordings of three broadly distributed frog species. There was no appreciable effect of anthropogenic habitat modification on any of the measured call variables, although there was high variability. With continued habitat modification, species may shift towards earlier and longer breeding seasons, with largely unknown ecological consequences in terms of proximate and ultimate fitness.

A new species of Leptobrachella (Anura, Megophryidae) from Mount Pu Ta Leng, northwest Vietnam.

The Hoang Lien Range in northwest Vietnam is known to harbour five species of Leptobrachella, a specious genus of terrestrial frogs. We collected specimens of Leptobrachella from Mount Pu Ta Leng on the second highest peak in the Hoang Lien Range and use morphological and molecular data to show that this population represents a previously undescribed species which we name Leptobrachella graminicola sp. nov. after the abundance of calling males on sedge-like plants. This new species is closely related to L. bourreti but the new species is distinguished from L. bourreti and other congeneric species by a combination of the following characters: (1) a body size range of 23.124.6 mm in six adult males and 28.632.9 mm in five adult females; (2) skin on dorsum smooth, with many tubercles and lacking dermal ridges; (3) toes with rudimentary webbing and broad lateral fringes; (4) belly white with brown spots; (5) throat dark brown with light grey-brown flecks and spots; (6) a row of large white spots on the outer edge of the tarsus extending from the heel to the inner metatarsal tubercle, sometimes forming a long white stripe; and (7) a bicoloured iris. The new, high-elevation species is likely to be range-restricted and threatened by habitat loss and the activities of tourists.

Amolops caelumnoctis Rao amp; Wilkinson, 2007, a Junior Synonym of A. splendissimus Orlov amp; Ho, 2007 (Amphibia: Anura: Ranidae).

Amolops splendissimus Orlov and Ho, 2007 and A. caelumnoctis Rao and Wilkinson, 2007 were described almost simultaneously from either side of the China-Vietnam border. The two species share a strong morphological resemblance, and their taxonomic distinctiveness has been questioned, yet no one has confirmed the taxonomic relationship and status between the two taxa. To resolve this taxonomic issue, we collected additional topotypic and near-topotypic specimens of A. splendissimus and A. caelumnoctis from both China (A. caelumnoctis: Wenshan County, Yunnan Province; type locality Luchun County, Yunnan Province), and Vietnam (A. splendissimus: Tam Duong District, Lai Chau Province; type locality Mount Ky Quan San, Bat Xat, Lao Cai Province). Molecular analysis based on a 16S rRNA fragment revealed minimal genetic divergences between the two taxa (0.0%0.4% uncorrected p-distance), and both species are closely related to A. viridimaculatus (2.1%2.3%) and A. medogensis (3.5%3.7%). Morphological comparisons between the newly collected specimens and the original descriptions of both species further support the lack of distinctiveness of the two species, hence, we conclude that A. caelumnoctis is a junior synonym of A. splendissimus.

Amphibians in Zootaxa: 20 years documenting the global diversity of frogs, salamanders, and caecilians.

Zootaxa is a mega-journal that since its inception, 20 years ago, has contributed to the documentation of the planet's biodiversity. Its role concerning terrestrial vertebrates has been crucial especially for amphibians, which are the most threatened class of vertebrates. As current editors of the Amphibia section, we reviewed the state of knowledge of taxonomic publications on amphibians over the last two decades (from 2001 to 2020). Our review reveals that 2,533 frogs, 259 salamanders, and 55 caecilians have been named in these 20 years, mainly in the tropical regions of South America, Asia, and Africa. More than half (57%) of these species descriptions were published in only 10 journals. At least 827 species of the new amphibians (29% of the total) were described in Zootaxa. This mega-journal has served also as a place of publication for monographs and systematic reviews, in addition to short articles documenting the vocalizations of anurans and the morphology of embryos and larvae. Its efficient evaluation process, the freedom of manuscript length, including full-color figures, and free of cost for the authors, has made Zootaxa a favorite for amphibian researchers. In an era of accelerating rates of biodiversity loss, documenting, describing, naming, and proposing evolutionary scenarios for species is, more than ever, an urgent task.

Species' traits drive amphibian tolerance to anthropogenic habitat modification.

Anthropogenic habitat modification is accelerating, threatening the world's biodiversity. Understanding species' responses to anthropogenic modification is vital for halting species' declines. However, this information is lacking for globally threatened amphibians, informed primarily by small community-level studies. We integrated >126,000 verified citizen science observations of frogs, with a global continuous measure of anthropogenic habitat modification for a continental scale analysis of the effects of habitat modification on frogs. We derived a modification tolerance index-accounting for anthropogenic stressors such as human habitation, agriculture, transport and energy production-for 87 species (36% of all Australian frog species). We used this index to quantify and rank each species' tolerance of anthropogenic habitat modification, then compiled traits of all the frog species and assessed how well these equipped species to tolerate modified habitats. Most of Australia's frog species examined were adversely affected by habitat modification. Habitat specialists and species with large geographic range sizes were the least tolerant of habitat modification. Call dominant frequency, body size, clutch type and calling position (i.e. from vegetation) were also related to tolerance of habitat modification. There is an urgent need for improved consideration of anthropogenic impacts and improved conservation measures to ensure the long-term persistence of frog populations, particularly focused on specialists and species identified as intolerant of modified habitats.

Anatomical features of the phytotelma dwelling, egg-eating, fanged tadpoles of Rhacophorus vampyrus (Anura: Rhacophoridae).

Tadpoles of the Vampire tree frog Rhacophorus vampyrus differ substantially from other rhacophorid tadpoles, by having profound modifications in external morphology. The morphological peculiarities of this species likely correlate with their arboreal microhabitat and strict oophagous diet. In this work, we examine buccal and musculoskeletal anatomy and compare them to other rhacophorid and egg-eating larvae. The shape and arrangement of cartilages of the lower jaw are unique among tadpoles, and the lack of a palatoquadrate suspensorium is only known in the distantly related macrophagous tadpoles of the dicroglossid Occidozyga baluensis. The cranial musculature is massive, and the morphology of several mandibular, hyoid, and abdominal muscles could be related to the ingestion and transit of large eggs. In the buccal cavity, conspicuous aspects are the absence of ridges and papillae, and the development of a unique glandular zone in the buccal floor. Finally, observations of the skeletal support of keratinized mouthparts allow us to present a topography-based hypothesis of homology of the conspicuous fangs of these tadpoles.

Bad neighbours: amphibian chytrid fungus Batrachochytrium dendrobatidis infection dynamics in three co-occurring frog species of southern Sydney, Australia.

Wildlife disease is a major cause of global biodiversity loss. Amongst the most devastating is the disease chytridiomycosis, caused by the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd). This disease has contributed to declines and extinctions in hundreds of amphibian species, but not all species are affected equally. Some amphibian hosts are capable of carrying high levels of Bd infection without population declines, acting as reservoir species for the pathogen and driving population declines in sympatric species. In Australia, several species have been proposed as reservoir species; however, our understanding of Bd is derived from studies that are highly geographically and taxonomically biased, and our ability to extrapolate from these systems is unknown. We examined the prevalence and intensity of Bd infection in 3 frog species in a previously unstudied host-pathogen system in temperate eastern Australia: the Blue Mountains tree frog Litoria citropa, a poorly-known species predicted to be susceptible to Bd infection; and the common eastern froglet Crinia signifera and the stony creek frog L. lesueuri, which have both been identified as reservoir species in other regions. We found that L. citropa and L. lesueuri were infected with Bd at a high prevalence and often high intensity, while the reverse was true for C. signifera. All species were detected at moderate abundance and there was no evidence of morbidity and mortality. Our findings do not support C. signifera and L. lesueuri being reservoir species in this system, highlighting the importance of region-specific studies to inform conservation management.

A new fanged frog in the Limnonectes kuhlii complex (Anura: Dicroglossidae) from northeastern Cambodia.

The Limnonectes kuhlii complex is a group of morphologically similar species of fanged frogs distributed across much of mainland and insular Southeast Asia. Many new species in this complex have been described in recent years, primarily on the basis of mitochondrial DNA divergence corroborated by differences in linear measurements and qualitative characters. Males in this species complex develop enlarged heads at sexual maturity, but the degree of head enlargement varies among mature males, even within the same population. We evaluated the utility of body length (snout-vent length minus head length) in descriptive statistics and in size-adjusting measurements for traditional morphometric analysis, as well as a landmark-based geometric morphometric analysis of male head shape, in Indochinese species of the L. kuhlii complex. The analyses supported quantitative and qualitative morphological distinction of a divergent mitochondrial lineage of the L. kuhlii complex in northeastern Cambodia, and the lineage is described as a new species. Limnonectes fastigatus sp. nov. differs from its closest relatives and from geographically proximate members of the complex by having the combination of elongated, slender odontoids; nuptial pads on the first finger; immaculate belly; significantly different body length-adjusted measurements in both sexes; and a significantly different male head shape. The new species is the only member of the L. kuhlii complex known from Cambodia.

Anuran accents: Continental-scale citizen science data reveal spatial and temporal patterns of call variability.

Many animals rely on vocal communication for mating advertisement, territorial displays, and warning calls. Advertisement calls are species-specific, serve as a premating isolation mechanism, and reinforce species boundaries. Nevertheless, there is a great deal of interspecific variability of advertisement calls. Quantifying the variability of calls among individuals within a species and across species is critical to understand call evolution and species boundaries, and may build a foundation for further research in animal communication. However, collecting a large volume of advertisement call recordings across a large geographic area has traditionally posed a logistical barrier. We used data from the continental-scale citizen science project FrogID to investigate the spatial and temporal patterns of call characteristics in six Australian frog species. We found intraspecific call variability in both call duration and peak frequency across species. Using resampling methods, we show that variability in call duration and peak frequency was related to the number of individuals recorded, the geographic area encompassed by those individuals, and the intra-annual time difference between those recordings. We conclude that in order to accurately understand frog advertisement call variation, or "anuran accents," the number of individuals in a sample must be numerous (N ≥ 20), encompass a large geographic area relative to a species' range, and be collected throughout a species' calling season.

-A new species of phytotelm breeding frog (Anura: Rhacophoridae) from the Central Highlands of Vietnam.

We describe a new species of phytotelm-breeding rhacophorid frog from central Vietnam. Gracixalus trieng sp. nov. is distinguished from all congeners by a combination of (1) body size medium (37.2-41.4 mm in five adult males), (2) snout rounded in dorsal and lateral views, (3) dorsal surface brown or yellowish with a darker brown interorbital crossbar and inverse-Y shape on the back, (4) throat and chest yellow or yellowish brown with pinkish mottling and belly and ventral surfaces of limbs including hands and feet pinkish, (5) tympanum and supratympanic fold distinct, (6) iris pale gold with darker gold radiating out from anterior and posterior edges of pupil, (7) majority of dorsal body and limb surfaces smooth in adults, with some individuals having sparsely distributed low, irregular tubercles, (8) nuptial pads on fingers I and II in adult males, and (9) eggs deposited as a tightly spaced array of non-pendent eggs on the wall of a phytotelmon. The species occurs in syntopy with G. lumarius. At present, Gracixalus trieng sp. nov. is known only from montane bamboo and evergreen forest (>1700 m) on Mount Ngoc Linh and adjacent peaks; and it is likely to be restricted to high-elevation forest with an estimated geographical distribution of <1000 km2.

A new Leptobrachella (Anura: Megophryidae) from the Cardamom Mountains of Cambodia.

The species-rich, megophryid frog genus Leptobrachella Smith, 1925 is distributed throughout the uplands of mainland Southeast Asia but is conspicuously absent from the Cardamom Mountains of southwestern Cambodia, where it has been known only by a single, undetermined metamorphic specimen collected nearly two decades ago. We report two adult female specimens of Leptobrachella collected a decade later at a second locality in the Cardamom Mountains and use mitochondrial DNA, nuclear DNA, and morphology to show that it represents a distinct species. Leptobrachella neangi sp. nov., named after its collector, is most closely related in mitochondrial and nuclear DNA to L. fuliginosa (Matsui, 2006) and L. melanoleuca (Matsui, 2006) from western Thailand, but has uncorrected pairwise distances of 8.69-10.99% in a mitochondrial 16S gene fragment from its two sister species. The new species is also readily distinguished from these and other congeners by having the combination of (1) SVL 35.4-36.3 mm in two adult females, (2) distinct dorsolateral glandular line absent, (2) belly transparent, immaculate purplish gray in life, creamy white in preservative, (3) dark inguinal blotch absent, (4) tympanum with black coloration extending from line under supratympanic fold, (5) dorsal skin with small, irregular bumps and ridges, and (6) iris coppery orange around pupil, fading to gold at periphery, not distinctly bicolored. The new species is the first named Leptobrachella from the Cardamom Mountains and the third from Cambodia.

A description of the tadpole of the Critically Endangered Botsford's leaf-litter frog (Leptobrachella botsfordi) with comments on the distribution and conservation status of the species.

We describe the tadpole of the Botsford's leaf-litter frog Leptobrachella botsfordi, a species endemic to Mount Fansipan in northwest Vietnam, for the first time. Tadpoles of this species were found in steep, fast-flowing streams at elevations between 2500-2600 m asl. We also report a previously unknown locality for this species on Mount Fansipan, which increases the species' known Extent of Occurrence from 8 km2 to 36 km2. Our findings will inform subsequent conservation initiatives for this poorly known and highly threatened species.

The tadpoles of five Megophrys Horned frogs (Amphibia: Megophryidae) from the Hoang Lien Range, Vietnam.

Frogs in the genus Megophrys are an Asian radiation of stream-breeding frogs. The tadpoles of many Megophrys species are undescribed; those that are described are often dubiously allocated to species by association with post metamorphic specimens at collection sites and without supportive molecular data. We provide detailed descriptions of the larvae of five species of Megophrys from the Hoang Lien Range in northwest Vietnam: Megophrys fansipanensis, M. gigantica, M. hoanglienensis, M. jingdongensis and M. maosonensis. Tadpoles from different subgenera differ from each other via a combination of patternation in life, oral disc shape and tail morphology but given the small sample size, and limited number of species it is unlikely that these differences can be applied more widely to delineate subgenera. Morphological differences between tadpoles from species within the subgenus Panophrys were insufficient to clearly delineate all species. The ability to identify tadpoles is likely to advance our understanding of the frog fauna in mainland southeast Asia.

The FrogID dataset: expert-validated occurrence records of Australia's frogs collected by citizen scientists.

This dataset represents expert-validated occurrence records of calling frogs across Australia collected via the national citizen science project FrogID (http://www.frogid.net.au). FrogID relies on participants recording calling frogs using smartphone technology, after which point the frogs are identified by expert validators, resulting in a database of georeferenced frog species records. This dataset represents one full year of the project (10 November 2017-9 November 2018), including 54,864 records of 172 species, 71% of the known frog species in Australia. This is the first instalment of the dataset, and we anticipate providing updated datasets on an annual basis.

Optimizing future biodiversity sampling by citizen scientists.

We are currently in the midst of Earth's sixth extinction event, and measuring biodiversity trends in space and time is essential for prioritizing limited resources for conservation. At the same time, the scope of the necessary biodiversity monitoring is overwhelming funding for professional scientific monitoring. In response, scientists are increasingly using citizen science data to monitor biodiversity. But citizen science data are 'noisy', with redundancies and gaps arising from unstructured human behaviours in space and time. We ask whether the information content of these data can be maximized for the express purpose of trend estimation. We develop and execute a novel framework which assigns every citizen science sampling event a marginal value, derived from the importance of an observation to our understanding of overall population trends. We then make this framework predictive, estimating the expected marginal value of future biodiversity observations. We find that past observations are useful in forecasting where high-value observations will occur in the future. Interestingly, we find high value in both 'hotspots', which are frequently sampled locations, and 'coldspots', which are areas far from recent sampling, suggesting that an optimal sampling regime balances 'hotspot' sampling with a spread across the landscape.