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Galápagos marine iguanas are primarily associated with the marine environment and show special nutritional adaptations. They are the only lizards worldwide that forage on marine macroalgae. Until now, consumed algae have been identified by direct observations during their feeding activities and microscopic identification in faeces samples. In this study, we use a novel DNA metabarcoding approach to identify consumed algal species from the faeces of marine iguanas. We developed primers for the ribulose-bisphosphate carboxylase (rbcL) gene and applied a metabarcoding approach to 25 individual faeces samples collected in four representative sites of two subspecies (Amblyrhynchus cristatus mertensi and A. c. godzilla), found in the San Cristóbal Island. We detected 18 consistently occurring macroalgal operational taxonomic units (OTUs). Most of the OTUs were assigned to Rhodophyta (red algae) and only one OTU to Chlorophyta (green algae). Despite the number of consumed algal species did not differ between two subspecies (OTU richness; P = 0.383), diet overlap level between A. c. mertensi and A. c. godzilla was low (Schoener index = 0.345), suggesting that both subspecies consumed different algal species in their natural environment. Further studies are needed to understand whether the difference of consumed algae reflects disparities in the abundance of algal species between sites, or whether iguanas of the two genetically differentiated subspecies prefer distinct algal species. [ABSTRACT FROM AUTHOR]

Epidemiology relies on understanding the distribution of pathogens which often can be detected through DNA-based techniques, such as quantitative Polymerase Chain Reaction (qPCR). Typically, the DNA of each individual sample is separately extracted and undergoes qPCR analysis. However, when performing field surveys and long-term monitoring, a large fraction of the samples is generally expected to be negative, especially in geographical areas still considered free of the pathogen. If pathogen detection within a population – rather than determining its individual prevalence – is the focus, work load and monetary costs can be reduced by pooling samples for DNA extraction. We test and refine a user-friendly technique where skin swabs can be pooled during DNA extraction to detect the amphibian chytrid fungi, Batrachochytrium dendrobatidis and B. salamandrivorans (Bsal). We extracted pools with different numbers of samples (from one to four swabs), without increasing reaction volumes, and each pool had one sample inoculated with a predetermined zoospore amount. Pool size did not reduce the ability to detect the two fungi, except if inoculated with extremely low zoospore amounts (one zoospore). We confirm that pooled DNA extraction of cutaneous swabs can substantially reduce processing time and costs without minimizing detection sensitivity. This is of relevance especially for the new emerging pathogen Bsal , for which pooled DNA extraction had so far not been tested and massive monitoring efforts in putatively unaffected regions are underway. [ABSTRACT FROM AUTHOR]

In the face of the global biodiversity crisis, the monitoring of species richness and diversity is experiencing an increased demand entailing a raise in cost and time investment. The analysis of species-specific DNA fragments in environmental samples (eDNA) such as from water or soil, facilitate the molecular detection of species without the specific sampling of individuals. The invasive chytrid fungus Batrachochytrium salamandrivorans (Bsal) is infecting natural fire salamander populations (Salamandra salamandra) and causes chytridiomycosis resulting in infrequent regional extinctions of populations across Central Europe. With regard to the expanding distribution of Bsal over the last years, cost-effective monitoring of fire salamanders is important for the conservation of this species. Based on a real-time quantitative PCR (qPCR) assay, we developed a new protocol to detect S. salamandra larvae in streams via eDNA, using species-specific primers of the mitochondrial control region (D-loop). We tested the efficiency of qPCR primer sets for six combinations of DNA extraction kits coupled with subsequent PCR inhibitor removal kits for obtaining qPCR-detectable S. salamandra eDNA from water filters, that were taken both from natural streams and artificial water tanks in the laboratory as positive controls. We found that the DNeasy Blood & Tissue Kit in combination with the DNeasy PowerClean CleanUp Kit performed best for detecting salamander larvae from natural streams. Our experimental protocol paves the way for resource-saving approaches to monitor S. salamandra larvae, but also confirms the limits to this eDNA approach in that it requires optimized laboratory protocols. [ABSTRACT FROM AUTHOR]

The North African fire salamander, Salamandra algira, is distributed in Algeria, Morocco and Ceuta (Spanish territory located on the north coast of Africa), but until now rather limited information has been available on the populations across the Algerian part of its range. We here provide a first analysis of the phylogeography of this species in Algeria, based on 44 samples from populations distributed across 15 localities in Central Algeria. We sequenced three segments of mitochondrial DNA, covering 12S rRNA, cytochrome b (Cytb) and the D-loop. The mtDNA sequences of the Algerian samples were strongly different from the Moroccan populations occurring west of the Moulouya River (corresponding to the subspecies S. a. tingitana and S. a. splendens) but sister to the genetically rather similar population from the Beni Snassen Massif in eastern Morocco (subspecies S. algira spelaea). Among the Algerian specimens studied, those from the westernmost site, Chrea Massif, were the sister clade to the remaining populations, and the overall genetic divergence was low, with a maximum of five mutational steps in a 295 bp fragment of cytochrome b. [ABSTRACT FROM AUTHOR]

We assessed the genetic variation of 47 Moroccan populations of the North African water frog (Pelophylax saharicus) using partial sequences of the mitochondrial cytochrome oxidase subunit I gene (COI). All 239 samples belonged to the main haplotype clade previously identified from Morocco, with no haplotypes of the Algerian/Tunisian clades among our samples. Altogether 40 haplotypes were found, with a maximum of 13 mutational steps between them. Two weakly divergent haplogroups separated by a minimum of six mutational steps were distributed (i) in the Anti-Atlas and the Sahara (south of the High Atlas Mountains) versus (ii) in the Middle Atlas, the High Atlas, and in the Rif area north of the Atlas Massif. Haplotypes of the northern haplogroup were found at the southernmost locality, which might be due to human translocation, and co-occurrence of the two haplogroups was recorded at three sites within the range of the northern haplogroup. [ABSTRACT FROM AUTHOR]

The emerging infectious disease chytridiomycosis is one of the major factors triggering global amphibian declines. A recently discovered species of chytrid fungus, Batrachochytrium salamandrivorans (Bsal), likely originated in East Asia, has led to massive declines in populations of fire salamanders (Salamandra salamandra) after its apparent introduction to the Netherlands and Belgium. Here, we report the first detection of this pathogen in Germany where it caused mass mortality of fire salamanders in a captive collection. Salamanders from this collection showed an almost 100% prevalence of infection with Bsal. Supposed Bsal -induced mortality occurred in multiple Salamandra species (S. salamandra , S. algira , S. corsica , and S. infraimmaculata), while Bsal infection was confirmed in nine subspecies of S. salamandra and in S. algira. Our study indicates that this pathogen can potentially infect all fire salamander species and subspecies. If Bsal spreads from captive collections to wild populations, then a similar devastating effect associated with high mortality should be expected. [ABSTRACT FROM AUTHOR]

Amphibians vary in the degree of pre-metamorphic developmental plasticity in response to risk of predation. Changes in hatching time and development rate can increase egg or tadpole survival respectively by shortening the duration of the more vulnerable stages. The intensity of predator induced developmental response and its direction, i.e. delayed, accelerated, or none, varies considerably between amphibian and predator species. We surveyed freshly deposited clutches of the European common frog Rana temporaria in a population in Braunschweig, Germany and found that 62% (N = 20) of the clutches contained planarians (Schmidtea nova), with an average of 3.94 ± 0.79 and a maximum of 13 planarians per clutch. A laboratory predation experiment confirmed that this planaria preys on R. temporaria eggs and early embryos. We further exposed freshly laid egg masses to either free, caged, or no planarians treatments using floating containers within a breeding pond where the two species co-occur. After 10 days exposure, embryos showed developmental stages 14-25 along the Gosner scale with statistically significant positive effects of both predator treatments. The observed effect was rather slight as predator-exposed individuals showed an increase by a single Gosner stage relative to those raised without planarians. The detected trend suggests that direct and indirect cues from flatworms, rarely considered as anuran predators, might induce a developmental response in R. temporaria early developmental stages. [ABSTRACT FROM AUTHOR]

The mucous skin of amphibians provides a habitat for microorganisms which may interact with their hosts and thereby affect their condition and health. Cultivation-independent analyses of the bacterial communities based on the detection of PCR-amplified bacterial 16S rRNA genes provides a direct approach to characterize their diversity. In the present pilot study we utilized this approach in combination with a high-throughput DNA sequencing technology (454 pyrosequencing), to characterize the bacterial community structure of the skin of three newt species (Lissotriton vulgaris, Ichthyosaura alpestris, Triturus cristatus), collected near Braunschweig, Germany. 16S rDNA sequences were obtained from 19 unique samples. On average, 6113 amplicon sequences were obtained per sample and these could phylogenetically be assigned to a total of 1615 different operational taxonomic units (OTUs). Altogether, most samples were rather similar in their dominant bacterial taxa. Most represented were Betaproteobacteria (46%; mostly Janthinobacterium), Gammaproteobacteria (28%; mostly Pseudomonas), Flavobacteria (phylum Bacteroidetes: 19%, mostly Flavobacterium), and Sphingobacteria (Bacteroidetes: 5%, mostly Pedobacter). We found no significant differences between the three newt species, or between hemi-nested vs. non-nested PCR, but a strong difference among sampling dates (15 and 17 April 2013) which might be explained by the ongoing transition of the newts from their terrestrial to aquatic phase which coincided with this period, or by differences between sexes as these were unevenly sampled on the two dates. 16S rRNA gene sequences retrieved in this study in several cases were identical or very similar to those previously found on the skin of North American salamanders. [ABSTRACT FROM AUTHOR]

Painted frogs (Discoglossus) contain five to six species of Western Palearctic anurans that are mainly distributed in allopatry. We here provide the first comprehensive assessment of the phylogeography of the Moroccan species D. scovazzi and geographically characterize its contact zone with D. pictus in Eastern Morocco. Discoglossus scovazzi shows, in general, a weak phylogeographic structure across Morocco on the basis of mitochondrial DNA sequences of the cytochrome b gene, with only populations centered in the Atlas Mountains characterized by the presence of slightly divergent haplotypes. In eastern Morocco, all populations east of the Moulouya River were clearly assignable to D. pictus. This species was also found along the Mediterranean coast west of the Moulouya, in the cities of Nador and Melilla, suggesting that not the river itself but the wide arid valley extending along much of the river (except close to the estuary) acts as a possible distributional barrier to these frogs. No sympatry of D. scovazzi with D. pictus was observed, and all specimens were concordantly assigned to either species by DNA sequences of cytochrome b and of the nuclear marker RAG 1. Species distribution models of the two taxa show largely overlapping areas of suitable habitat, and the two species' niches are significantly more similar than would be expected given the underlying environmental differences between the regions in which they occur. Comparative data are also presented from the southern Iberian contact zone of D. galganoi galganoi and D. g. jeanneae. These taxa showed less clear-cut distributional borders, extensively shared RAG1 haplotypes, and had instances of sympatric occurrence on the basis of cytochrome b haplotypes, in agreement with the hypothesis of a yet incomplete speciation. In this wide contact zone area we found mitochondrial sequences containing double peaks in electropherograms, suggesting nuclear pseudogenes or (less likely) heteroplasmy, possibly related to the ongoing admixture among the lineages. [ABSTRACT FROM AUTHOR]

We provide a DNA barcoding survey of Malagasy amphibians, including 251 of the 292 nominal species known to date, by complementing previous data with 280 newly determined barcoding sequence fragments of the mitochondrial cytochrome oxidase subunit I (COI) gene. Amplification success for the newly determined sequences was highest (94%) with one set of universal COI primers (dgLCO1490-dgHCO2198) while other primer sets had distinctly lower success rates. By and large, we observed relatively high average interspecific genetic distances of 25-27% within the Mantellidae and Microhylidae, and genetic distances of 13-21% within the Hyperoliidae. Lower values of 6-7% were observed between some sister species in all families, with extreme lows of 0.2-0.3% between a few sister species pairs in microhylids and mantellids for which we postulate mitochondrial introgression or yet unsettled taxonomy. Within-species divergences were relatively high especially in mantellids where they averaged 5.3%, due to the inclusion of numerous deep conspecific lineages (by definition with high divergences to other specimens) in our study. Above this, the degree of polymorphism was difficult to establish owing to limited sampling per population in our assessment. Compared to a previous assessment from 2009 based on 16S rDNA sequences, we identify 14 additional undescribed candidate species and raise the maximum estimate of species in the island's batrachofauna to well over 500. [ABSTRACT FROM AUTHOR]

A precise knowledge of the spatial distribution of taxa is essential for decision-making processes in land management and biodiversity conservation, both for present and under future global change scenarios. This is a key base for several scientific disciplines (e.g. macro-ecology, biogeography, evolutionary biology, spatial planning, or environmental impact assessment) that rely on species distribution maps. An atlas summarizing the distribution of European amphibians and reptiles with 50 x 50 km resolution maps based on ca. 85 000 grid records was published by the Societas Europaea Herpetologica (SEH) in 1997. Since then, more detailed species distribution maps covering large parts of Europe became available, while taxonomic progress has led to a plethora of taxonomic changes including new species descriptions. To account for these progresses, we compiled information from different data sources: published in books and websites, ongoing national atlases, personal data kindly provided to the SEH, the 1997 European Atlas, and the Global Biodiversity Information Facility (GBIF). Databases were homogenised, deleting all information except species names and coordinates, projected to the same coordinate system (WGS84) and transformed into a 50 x 50 km grid. The newly compiled database comprises more than 384 000 grid and locality records distributed across 40 countries. We calculated species richness maps as well as maps of Corrected Weighted Endemism and defined species distribution types (i.e. groups of species with similar distribution patterns) by hierarchical cluster analysis using Jaccard's index as association measure. Our analysis serves as a preliminary step towards an interactive, dynamic and online distributed database system (NA2RE system) of the current spatial distribution of European amphibians and reptiles. The NA2RE system will serve as well to monitor potential temporal changes in their distributions. Grid maps of all species are made available along with this paper as a tool for decision-making and conservation-related studies and actions. We also identify taxonomic and geographic gaps of knowledge that need to be filled, and we highlight the need to add temporal and altitudinal data for all records, to allow tracking potential species distribution changes as well as detailed modelling of the impacts of land use and climate change on European amphibians and reptiles. [ABSTRACT FROM AUTHOR]

Based on concordant differences in male advertisement call, tadpole morphology, and absence of haplotype sharing in mitochondrial and nuclear DNA markers, we describe a new species of treefrog from Ranomafana National Park in the southern central east of Madagascar. In its adult stage Boophis narinsi sp. n. is highly similar to its sister species, Boophis majori, but appears to differ in having longer hindlimbs. The genetic divergences between these two species (2.5-3.3% in a fragment of the 16S rRNA gene, depending on fragment length and individual haplotype analyzed) are below the threshold typically characterizing distinct species of anurans. Together with their relatively small and largely overlapping ranges and their sympatric occurrence in Ranomafana National Park, this indicates that they potentially could have originated rather recently by adaptive speciation under parapatric or sympatric conditions. Most studies on amphibian speciation have so far by default assumed vicariant speciation. We suggest that alternative speciation scenarios should be considered in future works and characterize settings in which more reliable assessments of adaptive parapatric or sympatric speciation could be carried out. [ABSTRACT FROM AUTHOR]

Based on DNA sequences of fragments of the mitochondrial COI and 16S rRNA genes we provide a first assessment of genetic variability of Iberian treefrog populations that have been historically allocated to Hyla arborea, partly as subspecies H. a. molleri. Our data from 147 specimens and 33 populations confirm strong divergence between these frogs and Central European H. arborea but relatively low differentiation across their range, supporting their status as a separate species, H. molleri. Preliminary phylogeographic data indicate a possible weak genetic differentiation of populations from the northern coast of the region of Galicia. We suggest inclusion of nuclear markers and an extension of the sampling into the coastal regions of Asturias and Cantabria, as well as the identification of the contact zone between H. molleri and H. arborea in either the Spanish Basque country or in France, as priorities for future research on this species. [ABSTRACT FROM AUTHOR]

Age structure of populations of four species of endemic Malagasy frogs of the genus Mantella (M. aurantiaca, M. baroni, M. bernhardi, M. madagascariensis) was examined by skeletochronology based on 96 specimens from nine different localities. In more than half of these (57%), no lines of arrested growth (LAGs) were found, and the number of LAGs recognized in the remaining specimens was mostly one, and probably two in three specimens. It is generally considered that each LAG corresponds to one year of life; our results therefore confirm that in Mantella populations almost all specimens are in their first or second year of life. [ABSTRACT FROM AUTHOR]

We analysed the DNA sequences of the mitochondrial cytochrome b gene for 49 specimens of the Western spadefoot toad, Pelobates cultripes, from Spain, and one sample from France, with a focus on populations from Galicia in northwestern Spain. Our results point to a generally low mitochondrial differentiation of the species across its distributional range although some phylogeographic structure was detected. Galician populations from coastal localities, from the A Limia region, and from Monforte appeared to be closely related, whereas the only available sample from the relict Galician population from near O Barco de Valdeorras has the same haplotype as the samples from Burgos, in central Spain. [ABSTRACT FROM AUTHOR]

We provide a partial revision of the microhylid frogs of the genus Anodonthyla, endemic to Madagascar, based on comprehensive molecular, bioacoustic and morphological data sets that include newly collected specimens from multiple localities. The molecular trees provide strong evidence for the polyphyly of several nominal species as they were previously defined, especially of Anodonthyla boulengeri and A. nigrigularis. As a consequence, we here resurrect the nomen Mantella pollicaris Boettger as Anodonthyla pollicaris from the synonymy of A. boulengeri, and we describe four new species, all with strong genetic divergences to other nominal species: Anodonthyla emilei from Ranomafana National Park, a comparatively medium-sized species characterized by a multi-note advertisement call with high note repetition rate; A. theoi from Manombo Special Reserve, a small species characterized by low note repetition rate, long note duration and high spectral call frequency; A. vallani, a medium-sized species from Ambohitantely Special Reserve, characterized by low note repetition rate, long note duration and low spectral call frequency; and A. jeanbai, a small species from Andohahela National Park, characterized by a long and narrow head, presence of short dorsolateral folds, a very short first finger, and a yellowish ventral colour. A further candidate species comprises populations previously assigned to A. boulengeri from the Ranomafana region, which we do not describe because the corresponding data set is too fragmentary, and we refer to it as A. sp. aff. boulengeri 'Ranomafana'. The molecular phylogeny indicates recurrent shifts between high and low note repetition rates in calls, based mainly on three strongly supported sister groups: A. moramora with low repetition rate and A. nigrigularis with moderately low repetition rate; A. theoi with low repetition rate and A. pollicaris with high repetition rate; and A. vallani with low repetition rate and A. sp. aff. boulengeri 'Ranomafana' with high repetition rate. The two species with the northernmost ranges, A. hutchisoni and A. boulengeri, are phylogenetically nested within clades of species occurring further south, confirming that the center of origin of the genus Anodonthyla was most likely in the South East of Madagascar. [ABSTRACT FROM AUTHOR]

We describe a new treefrog species of the genus Boophis (subgenus Sahona) from eastern Madagascar. Boophis calcaratus sp. nov. is distinguished from other species of the group by the presence of distinct dermal heel tubercles which are especially well developed in females. We also re-examined the type specimens of Boophis hillenii, a species that has not been rediscovered after its description in 1979. According to our analysis this taxon shows no consistent morphological differences to Boophis idae and supposed bioacoustic differences may be explained by two distinct call types in this species, and we thus suggest to consider hillenii as junior synonym of idae. [ABSTRACT FROM AUTHOR]

We determined DNA sequences of the mitochondrial cytochrome b gene for 44 specimens of the common frog, Rana temporaria parvipalmata, from 14 localities in Galicia, northwestern Spain. We found a low differentiation among all studied individuals, with a maximum of four mutational steps between haplotypes, including those from the southernmost isolated populations on the Manzaneda-Queixa and Trevinca-Segundeira massifs which thus probably originated by a rather recent colonization and likely maintained intermittent gene flow to more northern populations during climatically colder, glacial periods with a higher habitat connectivity. [ABSTRACT FROM AUTHOR]

We describe Liophidium maintikibo, a new species of pseudoxyrhophiine snake from the dry deciduous forest of Kirindy, western Madagascar. The new species is related to Liophidium therezieni but differs by a lower number of ventral scales and several details of colouration. Its ventral scales have a large central black patch which is missing from the subcaudals. This predominantly black ventral side is similar to that of another enigmatic Malagasy snake, Pararhadinaea melanogaster, for which we here present evidence for two new locality records, Ankarana and Daraina in northern Madagascar. The phylogenetic relationships of P. melanogaster (the sole representative of a monotypic genus) have so far not been clarified. Similarities to Liophidium maintikibo may indicate affinities to Liophidium, but P. melanogaster differs from this new species by a lower number of ventrals, subcaudals, number of maxillary teeth and by a different dorsal colouration. [ABSTRACT FROM AUTHOR]

The iguanid subfamily Oplurinae consists of seven species of small to medium-sized, arboreal, sand-dwelling or rock-dwelling lizards endemic to Madagascar and the Comoros, belonging to the genera Oplurus and Chalarodon. We here present the first complete molecular species-level phylogeny for Oplurinae based on DNA sequences (865 bp) of the mitochondrial 16S rRNA gene and the nuclear c-mos gene. Our study is based on 52 specimens sampled from different populations in Madagascar and includes the geographically isolated population from Grande Comore, hitherto considered as subspecies Oplurus cuvieri comorensis or even as a separate species O. comorensis. Our results confirm that, within the genus Oplurus, the largely arboreal O. cuvieri and O. cyclurus form the sister clade to the remaining, more rock-dwelling species. Within the latter lineage, Oplurus quadrimaculatus is placed most basal, O. fierinensis and O. grandidieri are closely related sister species with high support, and O. saxicola is sister to the fierinensis/grandidieri lineage. Within the arboreal Oplurus, the Comoran sample shows no genetic differentiation relative to O. cuvieri populations from the North West and Sambirano regions of Madagascar, indicating that this population should not be considered as a separate species. In the monotypic genus Chalarodon, we discovered deep genetic divergences among populations of C. madagascariensis indicating the presence of a previously unrecognized cryptic species and the need for taxonomic revision. [ABSTRACT FROM AUTHOR]

Describes the advertisement calls of Mantidactylus guttulatus. Diversity in the morphological and ecological adaptation of mantellid frogs; Temporal characteristics of calls by Mantidactylus guttulatus; Territorial behavior of male frogs.

The treefrog genus Boophis is represented in Madagascar by 50 described species. In this paper we describe two new cryptic species: B. liami sp. nov. and B. solomaso sp. nov. are similar to B. mandraka in morphology and colouration, but are clearly different in bioacoustic and genetic characters. The existence of many sibling species within Boophis shows the importance of detailed studies to clarify the distribution and the ecological requirements of the species in this genus. Les anoures arboricoles du genre Boophis sont représentés à Madagascar par 50 espèces. Dans cet article nous décrivons deux nouvelles espèces cryptiques : B. liami sp. nov. et B. solomaso sp. nov. ressemblent a B. mandraka par la coloration et la morphologie, mais ce distinguent clairement par la bioacoustique et le caractères génétiques. L'existence de beaucoup de "sibling species" dans le genre Boophis montre l'importance des études détailles pour qu'on puisse clarifier les questions concernant la répartition et les demandes écologiques des espèces de ce genre. [ABSTRACT FROM AUTHOR]

We describe a new species of the genus Mantidactylus found during field inventories in the north-eastern rainforests of Madagascar. Mantidactylus salegy sp. n. reaches a snout-vent length of 45-50 mm, has evident dorsolateral ridges and whitish spots on the upper jaws (mainly in females). The vocal sacs in males are distinctly visible and paired subgular, without strongly inflatable areas recognizable between the blackish lateral skin flaps on the throat. This frog exhibits a mosaic of characters previously thought to be typical for either the subgenera Gephyromantis or Phylacomantis. Therefore, it is a crucial discovery towards better understanding of the relationships between the taxa included in these two subgenera, supporting previous molecular results which indicated their paraphyly. The call of M. salegy consists of a rapid series of 8-11 rattling notes emitted at 1000-3500 Hz. We also provide karyological data for M. salegy, and for several related species, M. granulatus, M. leucomaculatus, M. moseri, M. redimitus, M. zavona, and M. pseudoasper (attributed to the subgenus Phylacomantis) and M. luteus (subgenus Gephyromantis). In terms of general chromosome morphology and structure, NOR localisation and heterochromatin characteristics, M. salegy shows similarities to M. redimitus, M. zavona and M. leucomaculatus, all included within Phylacomantis. Our data confirm that NOR position is a phylogenetically informative character. [ABSTRACT FROM AUTHOR]

We describe a new frog species of the Mantidactylus boulengeri group (Amphibia: Anura: Mantellidae) in the subgenus Gephyromantis from south-eastern Madagascar. It is morphologically similar to M. eiselti and M. thelenae but differs in its advertisement calls with a distinctly shorter note duration, and in its bilobate (not single) subgular vocal sac. Like M. eiselti and M. thelenae the new species Mantidactylus enki is predominantly diurnal and calling males do not aggregate close to water bodies, indicating direct development as it has been demonstrated previously in M. eiselti. The advertisement calls of all three species are described in detail. A comparison with a further pair of cryptic species with bilobate vs. single subgular vocal sac (Mantidactylus tschenki — M. cornutus) did not reveal any consistent pattern of call variation correlated with vocal sac structure. Therefore, sexual selection may also be considered to explain differences in the colour and external structure of vocal sacs among closely related species. [ABSTRACT FROM AUTHOR]

Dramatic global amphibian declines have recently led to an increased concern for many species of this animal class. The enigmatic Hula painted frog (Latonia nigriventer), the first amphibian to be declared extinct but unexpectedly rediscovered in 2011, has remained one of the rarest and most poorly understood amphibians worldwide. Gathering basic biological information on this species, along with an understanding of its disease-related threats remains fundamental for developing risk assessments and conservation strategies. Our surveys in recent years confirmed that L. nigriventer is a localised species with elusive habits. The species appears to follow an opportunistic breeding phenology and has a tadpole morphology similar to its well-studied sister group Discoglossus. However, the adults’ extended annual presence in the aquatic habitat is a major difference from species of Discoglossus. We detected the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), in northern Israel and on Hula painted frogs but did not observe any signs of chytridiomycosis in this species. Our preliminary data on aspects of the innate immunity of L. nigriventer suggest that the skin mucosome of this species contains antimicrobial peptides and a bacterial community differing from other syntopic frogs (Pelophylax bedriagae). The combined knowledge of both natural history and innate immunity of L. nigriventer provides valuable insights to direct future research and conservation management of this critically endangered frog species. [ABSTRACT FROM AUTHOR]

In the context of DNA Barcoding, sequences of standard marker genes for thousands and potentially millions of individuals and species are becoming available, requiring ever more efficient bioinformatic environments and software algorithms for analysis. We here present ExcaliBAR (Extraction, Calculation, Barcoding), a user-friendly software utility to facilitate one important initial step in DNA barcoding analyses, namely the determination of the barcoding gap between pairwise genetic distances among and within species, based on original distance matrices computed by MEGA software. In addition, the software is able to rename sequences downloaded via the standard user interfaces of public databases such as GenBank, without the need of developing and applying specific scripts for this purpose. [ABSTRACT FROM AUTHOR]

We provide detailed morphological descriptions of the tadpoles of Malagasy river bank frogs of the subgenera Ochihomantis and Maitsomantis (genus Mantidactylus, family Mantellidae), and data on relative abundance and habitat preferences of Ochthomantis species from Ranomafana National Park in southeastern Madagascar. Our study includes the tadpoles of six de- scribed and four undescribed candidate species. Eight of these larvae were previously unknown. Tadpoles were identified by DNA barcoding. Due to the very rudimentary taxonomic knowledge on Ochihomantis, we followed a 'reverse taxonomy' approach in which adult classification was to a great part determined on the basis of larval differences. By this procedure we even identified one candidate species whose adults remain still unknown. The majority of tadpoles in Ochthomantis and Maitsomantis have a rather similar body shape and they usually have similar habitat requirements. However, on the basis of the structure of their oral disk we identified three distinct groups: the first includes the femoralis-like tadpoles of Mantidactylus femoralis, M. ambreensis, M. zolitschka, M. argenteus, and of the candidate species named M. sp. 42, M. sp. 43 and M. sp. 47. They all have a reduced oral disk with poorly keratinized jaw sheaths and labial teeth. The mocquardi-like tadpoles of M. mocquardi and M. sp. 64 are placed in the second group and are characterized by a further reduction of oral disk structures, i.e. a complete lack of labial teeth. The third group includes only M. majori and is characterized by the transformation of the upper jaw sheath into three thorn-shaped projections. Based on a preliminary molecular phylogenetic analysis the reduction of keratinized oral structures in M. majori may have occurred convergently to that in M. mocquardi. The ecological data indicate that the tadpoles of the three most abundant species in Ranomafana (M. femoralis, M. ma)on and M. sp. 47) do not obviously differ in their choice of microhabitat although the differences in their oral structures indicate that they might use different food resources. They all show a preference for the stream areas with slow current and leaf litter substrate. [ABSTRACT FROM AUTHOR]