Bufo Viridis Research Articles

Herpetofauna of the Podkielecki Landscape Protection Area

Abstract The study was conducted in 2016-2017 in the Podkielecki Landscape Protection Area (area 26,485 ha). It was focused on the occurrence and distribution of amphibians and reptiles, the biology of the selected species and the existing threats. Established in 1995, the Podkielecki Landscape Protection Area surrounds the city of Kielce from the north, east and south-east, and adjoins several other protected areas. It covers the western part of the Świętokrzyskie Mountains (part of the Klonowskie and Masłowskie ranges) and the southern part of the Suchedniów Plateau. The studied area is mostly covered by forest and thicket communities (48.1%) and farmlands (39.9%), followed by built-up areas (7.8%), industrial areas (0.5%), roads and railways (2.7%), and surface water bodies (1%). The protected area is developed mainly on Palaeozoic rocks, including Cambrian and Ordovician sandstones, Silurian and Carboniferous shales, and Devonian marls. Podzolic soils predominate among soils. The largest rivers include Lubrzanka, Czarna Nida, Bobrza and Belnianka. There are no natural lakes within the PLPA limits, and the largest artificial reservoirs include the Cedzyna Reservoir, Morawica Reservoir, Suków Sandpit and two sedimentation reservoirs of the Kielce Power Plant. The area includes 2 nature reserves: Barcza and Sufraganiec. The following amphibian species were recognised during the investigations within the borders of the studied area: alpine newt Ichthyosaura alpestris Laur., great crested newt Triturus cristatus Laur., smooth newt Lissotriton vulgaris L., European fire-bellied toad Bombina bombina L., common spadefoot toad Pelobates fuscus Laur., common toad Bufo bufo L., natterjack toad Epidalea calamita Laur., European green toad Bufotes viridis Laur., European tree frog Hyla arborea L., pool frog Pelophylax lessonae Cam., edible frog Pelophylax esculentus L., marsh frog Pelophylax ridibundus Pall., moor frog Rana arvalis Nilss., and common frog Rana temporaria L. The reptiles were represented by sand lizard Lacerta agilis L., viviparous lizard Zootoca vivipara Jacquin, slow worm Anguis fragilis L., grass snake Natrix natrix L. and common European adder Vipera berus L. The study also included the phenology and breeding biology of the common toad and common frog. The most crucial herpetofauna conservation problems identified here include amphibians killed on roads by vehicles. The study area is intersected by very busy roads, in particular: European route no. E77, national roads nos. 73, 74 and S74, and regional roads nos. 745, 750 and 764. For this reason, future road reconstruction projects should consider the assembly of various crossing roads for wildlife, particularly on the 600 m long section of national road no. 74 near Cedzyna Reservoir. Other threats include illegal waste dumping, pollution of surface waters, fire setting, overgrowing and desiccation of small water bodies. © IOŚ-PIB

An unknown collection of lizards from Afghanistan.

Afghanistan is a herpetologically understudied country with few published papers since the end of “Afghanistan’s Golden Age” from the 1930s to the 1970s. Although a detailed checklist of the herpetofauna of the country, based on exploration of herpetodiversity using biodiversity archives, has been published recently, there still exist additional historical data that have not been considered. This is the case for a so far unknown collection of lizards from Afghanistan deposited in the herpetological collection of the Institute for Biological Research “Siniša Stanković at the University of Belgrade, Belgrade, Serbia. The material comes from field research conducted in 1972 and contains 27 specimens in seven lizard genera representing four families (Agamidae, Gekkonidae, Lacertidae, Scincidae). This historical collection was examined and basic morphometric data, field data, and photographs are provided, comparing the distributional data with published datasets. Updated species distribution maps reveal new locality or province records and an important range extension for Eurylepis taeniolata Blyth, 1854 which represents the northernmost record for this species in Afghanistan. In addition, one further distribution record for the Bufotes viridis (Laurenti, 1768) complex from the same research trip is noted.

Urban plums and toads: do fleshy fruits affect the post-metamorphic growth of amphibians?

BackgroundThe main aim of the study was to analyse the influence of fleshy fruits (plums) on the post-metamorphic growth and feeding behaviour of the green toad Bufotes viridis. We tested the following two hypotheses: (1) juveniles of the green toad are characterised by faster growth in conditions involving fallen plums Prunus cerasifera due to the associated presence of more varied food such as invertebrates; (2) green toads exhibit more active feeding behaviour in the presence of fleshy fruits.MethodsA total of 120 fresh metamorphs of the green toad were randomly assigned to one of four groups: two experimental groups with fleshy plums and two other groups as controls (without fruits). Each group was kept in an enclosure to which wild invertebrates had free access. Each individual toad was measured for snout-vent length (mm) and body mass (g) every other day for 30 days. In order to determine whether fallen plums influence the feeding behaviour of toads, the number of active and hidden (under an artificial shelter) individuals was also noted.ResultsThe results showed that green toads from both enclosures with plums were characterised by more rapid growth than individuals from the control treatments. Simultaneously, in the enclosure with fleshy fruits, greater species richness of wild invertebrates was observed. No differences in active feeding behaviour were noted between control groups and groups with plums.DiscussionFleshy fruits, upon falling, attract many types of invertebrates; thus they may represent good dietary supplements for fresh amphibian metamorphs. Therefore, the presence of fruit trees close to a breeding site might influences the post-metamorphic growth of amphibians, but not their feeding behaviour. The presence of insects associated with fallen fruit seems to favour the occurrence of amphibian populations, which is particularly important, since, due to political and social pressure, numbers of fruit trees are currently being reduced. To our knowledge, no previous study has investigated the potential influence of the presence of fruit trees on the growth and behaviour of anurans.

Morphological Deformities of Green Toad (Bufo viridis) Tadpoles Caused by Petroleum Products

Morphological Deformities of Green Toad (Bufo viridis) Tadpoles Caused by Petroleum Products

The effects of water and bottom sediments from Lake Balkash (Central Asia) on early development of fish and amphibians

The effects of water and bottom sediments from Lake Balkash (Central Asia) on early development of fish and amphibians

Hepatotoxicity and Nephrotoxicity of Lead Nitrate in Toad Bufo viridis

Hepatotoxicity and Nephrotoxicity of Lead Nitrate in Toad Bufo viridis

Introgression across Hybrid Zones Is Not Mediated by Large X-Effects in Green Toads with Undifferentiated Sex Chromosomes.

Divergence between incipient species remains an incompletely understood process. Hybrid zones provide great research potential, reflecting natural organismal genomic interactions and gene evolution in a variety of recombinants over generations. While sex chromosomes are known evolutionary drivers of reproductive isolation, empirical population genetics has mostly examined species with heteromorphic sex chromosomes. We recently reported restricted introgression at sex-linked markers in an amphibian system with homomorphic sex chromosomes (Hyla), consistent with a large X-effect, designating a greater role of sex chromosomes in driving hybrid incompatibilities. Here, using a similar approach, we examined two hybrid zones of Palearctic green toads (Bufo viridis subgroup), involving several lineages that arose at different times and form secondary contacts. We find no evidence for differential introgression of sex-linked versus autosomal markers across both zones. This absence of large X-effects in Bufo indicates that, unlike in Hyla, hybrid incompatibilities may not result from the faster-heterogametic sex and faster-male aspects of Haldane's rule. The recent suppression of XY recombination in Hyla but not in Bufo may have driven greater divergence between Hyla sex chromosomes, causing stronger reproductive isolation. Alternatively, stronger linkage among Hyla's sex-linked markers could restrict introgression. We hypothesize that the degree of sex-specific recombination may condition the importance of homomorphic sex chromosomes in speciation.

Prooxidant effects of chronic exposure to deltamethrin in green toad Bufotes viridis.

Pesticide-induced oxidative stress, as one of mechanism of toxicity, has been a focus of toxicological research. However, there is a lack of data for certain pesticides-oxidative stress effects especially on terrestrial amphibians. This study evaluates the prooxidative effects of orally administered insecticide deltamethrin (DM) in some tissues of the terrestrial toad Bufotes viridis. Toads were randomly divided and assigned to a control group and a test group that was orally exposed to the pesticide (5mg/kg of body weight/daily) for 21days. Animals were euthanized from each group on days 7, 14, and 21, and the liver, leg muscle, ventral skin, and gastrointestinal tissue (GIT) were dissected and used for analysis. From battery of investigated antioxidant components, superoxide dismutase (SOD) was the most differentiate parameter in all examined tissues. For the period of prolonged exposure to pesticide, antioxidative strategy of Bufotes viridis was based on SOD utilization in attempt to maintain the oxidative disbalance at acceptable level. The integrated biomarker response (IBR) as the measure of the overall biochemical response to DM exposure revealed that the group exposed for 21days had the highest response. Our work has offered valuable data ensuring evidence that toads exposed to deltamethrin developed adaptive reactions that were tissue-specific and that DM-generated systemic toxicity was time-dependent. The present work showed that oxidative stress has significant role in pesticide-induced toxicity and contributes to better understanding of ecotoxicological risk in the terrestrial amphibians exposed to DM.

Amphibians distribution and habitats in the southwestern region of Saudi Arabia.

This study surveyed 205 wetland sites in the Southwestern Saudi Arabia. We found seven species of amphibians (Anura), which belong to four families: Bufonidae; which includes four species, Sclerophrys tihamica; S. Arabica, Duttaphrynus dhufarensis, and Bufotes viridis; Hylidae, represented by only one species, the tree frog Hyla savignyi; Ranidae, which represented by one species, Pelophylax ridibundus and finally the Dicroglossidae, represented by one species, Euphlyctis ehrenbergii. The reasonable amount of rain received in southwest Arabia, and habitat diversity contribute to the relative abundance of amphibians in the region. Five types of wetland habitats were found in the study area. Valley streams, irrigated farms, seasonal ponds, dam reservoir and lagoons of treated sewage water. The current study revealed the wide spread of amphibians in southwestern Saudi Arabia than what had been reported earlier and confirmed the presence of the Bufotes viridis at three new sites in Asir Heights. It also showed the wide spread of S. arabica in all types of habitats in the southwestern region of Saudi Arabia. Habitat degradation was evident in the region, which might lead to species loss.

Phylogeography of Aegean green toads (Bufo viridis subgroup): continental hybrid swarm vs. insular diversification with discovery of a new island endemic

BackgroundDebated aspects in speciation research concern the amount of gene flow between incipient species under secondary contact and the modes by which post-zygotic isolation accumulates. Secondary contact zones of allopatric lineages, involving varying levels of divergence, provide natural settings for comparative studies, for which the Aegean (Eastern Mediterranean) geography offers unique scenarios. In Palearctic green toads (Bufo viridis subgroup or Bufotes), Plio-Pleistocene (~ 2.6 Mya) diverged species show a sharp transition without contemporary gene flow, while younger lineages, diverged in the Lower-Pleistocene (~ 1.9 Mya), admix over tens of kilometers. Here, we conducted a fine-scale multilocus phylogeographic analysis of continental and insular green toads from the Aegean, where a third pair of taxa, involving Mid-Pleistocene diverged (~ 1.5 Mya) mitochondrial lineages, earlier tentatively named viridis and variabilis, (co-)occurs.ResultsWe discovered a new lineage, endemic to Naxos (Central Cyclades), while coastal islands and Crete feature weak genetic differentiation from the continent. In continental Greece, both lineages, viridis and variabilis, form a hybrid swarm, involving massive mitochondrial and nuclear admixture over hundreds of kilometers, without obvious selection against hybrids.ConclusionsThe genetic signatures of insular Aegean toads appear governed by bathymetry and Quaternary sea level changes, resulting in long-term isolation (Central Cyclades: Naxos) and recent land-bridges (coastal islands). Conversely, Crete has been isolated since the end of the Messinian salinity crisis (5.3 My) and Cretan populations thus likely result from human-mediated colonization, at least since Antiquity, from Peloponnese and Anatolia. Comparisons of green toad hybrid zones support the idea that post-zygotic hybrid incompatibilities accumulate gradually over the genome. In this radiation, only one million years of divergence separate a scenario of complete reproductive isolation, from a secondary contact resulting in near panmixia.

First detection of the amphibian chytrid fungus (Batrachochytridium dendrobatidis) in free- living anuran populations in Greece

The amphibian chytrid fungus (Batrachochytrium dendrobatidis) is a widespread, cosmopolitan pathogen largely affecting free-living amphibian populations. So far there are no published studies for the presence of B. dendrobatidis in Greece. In this preliminary study we sampled 59 metamorphosed anurans from four Greek wetlands. Five samples were positive for the fungus by real-time PCR. B. dendrobatidis was detected in three species (Bufo viridis, Pelophylax epeiroticus, Pelophylax ridibundus) but not in endangered endemic Karpathos water frogs (Pelophylax cerigensis). This is the first report of the amphibian chytrid fungus (Batrachochytrium dendrobatidis) in free-living anuran populations from Greece.

Growth and development of the green toad (Bufo viridis) from the water bodies of the oil producing regions of Kazakhstan

Growth and development of the green toad (Bufo viridis) from the water bodies of the oil producing regions of Kazakhstan

Impacts of temperature on growth, development and survival of larvalBufo (Pseudepidalea) viridis(Amphibia: Anura): implications of climate change

The changing global climate poses a new threat to amphibian populations since amphibian growth and development are strongly influenced by temperature. In general, low temperature reduces their physiological processes, decreasing development and growth rates. However, metamorphs of some amphibian species may be larger than their conspecifics grown at high temperatures. The increased temperature can reduce the metamorphic size in some species, whereas in other species, high temperatures accelerate growth with no changes in body size at metamorphosis. In order to test the influence of water temperature on larval growth, development and survival of the green toad Bufo viridis, 200 eggs were raised separately at two different water temperatures (18 and 21 °C) for the period of 16 weeks. The results show that the larvae reared at high temperature reached metamorphosis significantly faster (86.58 ± 13.47 days) than those reared at low temperature (105.4 ± 7.83 days). However, the larvae reared at high temperature showed a significant increase in survival rate (64% ± 34.22) compared to the larvae reared at low temperature (23.57% ± 4.18). The growth rate of the larvae reared at higher temperature was higher than that of the larvae reared at low temperature but this difference was not significant. In contrast, the average size at metamorphosis (SVL) at low temperature was significantly larger (19.05 ± 1.12 mm) compared with that of the individuals that metamorphosed at the relatively higher temperature (16.11 ± 1.54 mm).

Cases of albinism and leucism in amphibians in Italy: new reports

Findings of abnormally pigmented amphibian individuals provide interesting insights on intraspecific phenotypic variability as well as on variation among populations inhabiting different habitats. Amphibian coloration is determined by chromatophores (specific epidermal cells), and a variety of abnormalities related to them have been reported. In this study we reported cases of albinism and leucism in six species of Italian amphibians, including some endemic species. For some taxa, like Hydromantes sarrabusensis, H. flavus, H. supramontis and Bufo viridis, we describe the first observations of albinism and leucism.

Neogene amphibians and reptiles (Caudata, Anura, Gekkota, Lacertilia, and Testudines) from the south of Western Siberia, Russia, and Northeastern Kazakhstan.

BackgroundThe present-day amphibian and reptile fauna of Western Siberia are the least diverse of the Palaearctic Realm, as a consequence of the unfavourable climatic conditions that predominate in this region. The origin and emergence of these herpetofaunal groups are poorly understood. Aside from the better-explored European Neogene localities yielding amphibian and reptile fossil remains, the Neogene herpetofauna of Western Asia is understudied. The few available data need critical reviews and new interpretations, taking into account the more recent records of the European herpetofauna. The comparison of this previous data with that of European fossil records would provide data on palaeobiogeographic affiliations of the region as well as on the origin and emergence of the present-day fauna of Western Siberia. An overview of the earliest occurrences of certain amphibian lineages is still needed. In addition, studies that address such knowledge gaps can be useful for molecular biologists in their calibration of molecular clocks.Methods and ResultsIn this study, we considered critically reviewed available data from amphibian and reptile fauna from over 40 Western Siberian, Russian and Northeastern Kazakhstan localities, ranging from the Middle Miocene to Early Pleistocene. Herein, we provided new interpretations that arose from our assessment of the previously published and new data. More than 50 amphibians and reptile taxa were identified belonging to families Hynobiidae, Cryptobranchidae, Salamandridae, Palaeobatrachidae, Bombinatoridae, Pelobatidae, Hylidae, Bufonidae, Ranidae, Gekkonidae, Lacertidae, and Emydidae. Palaeobiogeographic analyses were performed for these groups and palaeoprecipitation values were estimated for 12 localities, using the bioclimatic analysis of herpetofaunal assemblages.ConclusionThe Neogene assemblage of Western Siberia was found to be dominated by groups of European affinities, such as Palaeobatrachidae, Bombina, Hyla, Bufo bufo, and a small part of this assemblage included Eastern Palaearctic taxa (e.g. Salamandrella, Tylototriton, Bufotes viridis). For several taxa (e.g. Mioproteus, Hyla, Bombina, Rana temporaria), the Western Siberian occurrences represented their most eastern Eurasian records. The most diverse collection of fossil remains was found in the Middle Miocene. Less diversity has been registered towards the Early Pleistocene, potentially due to the progressive cooling of the climate in the Northern Hemisphere. The results of our study showed higher-amplitude changes of precipitation development in Western Siberia from the Early Miocene to the Pliocene, than previously assumed.

Testing for intraspecific and interspecific larval competition between two anurans: Hyla savignyi and Bufotes viridis

Intraspecific and interspecific competition can be important factors affecting anuran tadpoles in temporary pools. We focus on two co-occurring anuran species that are of conservation concern in Israel and may negatively interact: Bufotes viridis (Laurenti, 1768) and Hyla savignyi (Audouin, 1829). We conducted a replacement design mesocosm experiment to examine intraspecific and interspecific competition between tadpoles of these two species. Plastic tubs were assigned to five replacement design treatments: 0 Hyla + 20 Bufotes; 0 Hyla + 40 Bufotes; 20 Hyla + 20 Bufotes; 20 Hyla + 0 Bufotes; and 40 Hyla + 0 Bufotes, and a control with no tadpoles which was used to test for effects of the tadpoles on invertebrate community structure. While H. savignyi was significantly affected by intraspecific density, with smaller body mass and longer time to metamorphosis at the high-density treatment, B. viridis showed no intraspecific or interspecific density-dependent effects. However, B. viridis time to metamorphosis was negatively correlated with water temperature, while its survivorship was positively correlated with water temperature. The tadpoles affected neither abundance nor taxon richness of invertebrates. Our results suggest that stronger intraspecific competition than interspecific competition may contribute to the common co-occurrence of these two anurans in the same pools.

Cryopreservation of Bufotes viridis embryos by vitrification

Loss of biodiversity among amphibians is a current concern. Our hypothesis is that the embryos of amphibian species at risk of extinction could be cryopreserved by vitrification, using methods which have proved successful with fish oocyte. To test this hypothesis, samples of four cryoprotectants - methanol (MeOH), dimethyl sulphoxide (Me2SO), propylene glycol (PG) and polyethylene glycol (PEG), some singly, some in combination, were plunged in liquid nitrogen for 5 min to find the best solution for vitrification. To find the least toxic of these solutions, blastulae and stage G17 embryos of Bufotes Viridis, a typical amphibian, were exposed to solutions at different concentrations (0.5–10 M) for different lengths of time (15–30 min), with and without their normal protective jelly coats. In each case the number of survivors, which reached stage G25 was counted. Finally a series of embryos was vitrified in liquid nitrogen using the most efficient and least toxic cryoprotectants.Propylene glycol had the best vitrification characteristics, but MeOH vitrified at higher concentrations. The optimum regime, with the least toxic ctyoprotectants, consisted of 1M Me2SO for 15 min and a combination of 15% PEG(w/v) + 3M PG + 2M Me2SO for 3 min, with the jelly coat intact, followed by vitrification. This gave a survival percentage of 87.6% immediately after vitrification. Methods designed for cryopreservation of fish embryos make a good starting point for cryopreservation of the embryos of amphibian.

Impaired gonadal and somatic development corroborate vulnerability differences to the synthetic estrogen ethinylestradiol among deeply diverged anuran lineages

Amphibians are undergoing a global decline. One poorly investigated reason could be the pollution of aquatic habitats by endocrine disrupting compounds (EDCs). We tested the susceptibility to the synthetically stabilized estrogen 17α-ethinylestradiol (EE2) in three deeply diverged anuran species, differing in sex determination systems, types of gonadogenesis and larval ecologies. To understand whether data from the amphibian model Xenopus laevis (Pipidae) are analogous and applicable to only distantly related non-model amphibians, tadpoles of X. laevis, Hyla arborea (Hylidae) and Bufo viridis (Bufonidae) were simultaneously exposed to 50, 500 and 5000ng/L EE2 from hatching until completion of metamorphosis, using a flow-through-system under identical experimental conditions. Comparing molecularly established genetic with histologically assessed phenotypic sex in all species, we have recently shown that EE2 provoked numerous genetic-male-to-phenotypic-female sex reversals and mixed sex individuals, confirming overall its expected feminizing effect. In the present study, we focus on the influence of EE2 on gonadal and somatic development. Anatomy and histology revealed several species-specific effects. In both non-model species, H. arborea and B. viridis, high numbers of anatomically impaired gonads were observed. In H. arborea, exposed to 5000ng/L EE2, numerous underdeveloped gonads were detected. Whereas EE2 did not alter snout-to-vent length and body weight of X. laevis metamorphs, H. arborea showed a treatment-dependent decrease, while B. viridis exhibited an increase in body weight and snout-to-vent length. Apart from a concentration-dependent occurrence of yellowish skin color in several H. arborea, no organ-specific effects were detected. Since EE2 ubiquitously occurs in many aquatic ecosystems and affects sexual and somatic development, among EDCs, it may indeed contribute to amphibian decline. The inter-species variation in developmental EE2-effects corroborates species-specific vulnerability differences towards EDCs between deeply diverged amphibian groups.

The plasticizer bisphenol A affects somatic and sexual development, but differently in pipid, hylid and bufonid anurans

Due to their terrestrial habitats and aquatic reproduction, many amphibians are both very vulnerable and highly suitable bioindicators. The plasticizer bisphenol A (BPA) is one of the most produced chemical substances worldwide, and knowledge on its impacts on humans and animals is mounting. BPA is used for the industrial production of polycarbonate plastics and epoxy resins and found in a multitude of consumer products. Studies on BPA have involved mammals, fish and the fully aquatic anuran model Xenopus laevis. However, our knowledge about the sexual development of non-model, often semi-terrestrial anuran amphibians remains poor. Using a recently developed experimental design, we simultaneously applied BPA to two non-model species (Hyla arborea, Hylidae; Bufo viridis, Bufonidae) and the model X. laevis (Pipidae), compared their genetic and phenotypic sex for detection of sex reversals, and studied sexual development, focusing on anatomical and histological features of gonads. We compared three concentrations of BPA (0.023, 2.28 and 228 μg/L) to control groups in a high-standard flow-through-system, and tested whether conclusions, drawn from the model species, can be extrapolated to non-model anurans. In contrast to previous studies on fish and Xenopus, often involving dosages much higher than most environmental pollution data, we show that BPA causes neither the development of mixed sex nor of sex-reversed individuals (few, seemingly BPA-independent sex reversals) in all focal species. However, environmentally relevant concentrations, as low as 0.023 μg/L, were sufficient to provoke species-specific anatomically and histologically detectable impairments of gonads, and affected morphological traits of metamorphs. As the intensity of these effects differed between the three species, our data imply that BPA diversely affects amphibians with different evolutionary history, sex determination systems and larval ecologies. These results highlight the role of amphibians as a sensitive group that is responsive to environmental pollution.

A Single Transcriptome of a Green Toad (Bufo viridis) Yields Candidate Genes for Sex Determination and -Differentiation and Non-Anonymous Population Genetic Markers.

Large genome size, including immense repetitive and non-coding fractions, still present challenges for capacity, bioinformatics and thus affordability of whole genome sequencing in most amphibians. Here, we test the performance of a single transcriptome to understand whether it can provide a cost-efficient resource for species with large unknown genomes. Using RNA from six different tissues from a single Palearctic green toad (Bufo viridis) specimen and Hiseq2000, we obtained 22,5 Mio reads and publish >100,000 unigene sequences. To evaluate efficacy and quality, we first use this data to identify green toad specific candidate genes, known from other vertebrates for their role in sex determination and differentiation. Of a list of 37 genes, the transcriptome yielded 32 (87%), many of which providing the first such data for this non-model anuran species. However, for many of these genes, only fragments could be retrieved. In order to allow also applications to population genetics, we further used the transcriptome for the targeted development of 21 non-anonymous microsatellites and tested them in genetic families and backcrosses. Eleven markers were specifically developed to be located on the B. viridis sex chromosomes; for eight markers we can indeed demonstrate sex-specific transmission in genetic families. Depending on phylogenetic distance, several markers, which are sex-linked in green toads, show high cross-amplification success across the anuran phylogeny, involving nine systematic anuran families. Our data support the view that single transcriptome sequencing (based on multiple tissues) provides a reliable genomic resource and cost-efficient method for non-model amphibian species with large genome size and, despite limitations, should be considered as long as genome sequencing remains unaffordable for most species.