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This paper deals with a lifetime test of the hydraulic and transmission oil, Shell Spirax S4 ATF (Universal Tractor Transmission Oil (UTTO)). This fluid was used in the hydraulic and transmission circuit of a John Deere 5720 tractor. The fluid was assessed in terms of a possible contamination of chemical elements. Fluid samples were taken from a John Deere 5720 tractor at intervals of 250 engine hours. These samples were subjected to an IR spectroscopy analysis, a measuring of the kinematic viscosity at 40°C, and differential scanning calorimetry (DSC). The spectrometer, Spectroil Q100, has been used to monitor chemical elements. [ABSTRACT FROM AUTHOR]

This paper is focused on the impacts of the biodegradable hydraulic and transmission fluid (Universal Tractor Transmission Oil) on operation of tractor hydraulic and transmission systems. The fluid was used in the hydraulic and transmission systems of a Zetor Proxima 6321 tractor. Fluid samples were taken from the Zetor Proxima 6321 tractor at intervals of 250 engine hours. These samples were subjected to an infrared spectroscopy analysis in order to measure the total acid number and perform the ferrography analysis. This paper proves minimal impacts of the biodegradable hydraulic fluid on operation of the Zetor Proxima 6321 tractor. The biodegradable fluid meets the requirements for the operation of agricultural tractors in terms of low impacts on the wear of hydraulic components.

The aim of this article is to evaluate the technical condition of a mechanical four-speed gearbox tightly connected to a mechanical two-speed auxiliary gearbox placed in an off-road vehicle. When observing the technical condition of the mechanical gearbox, we used one of technical diagnostics methods, namely vibrodiagnostics. The mechanical gearbox was monitored during all its life cycle up to failure occurrence. In the article there is a detailed description of the stand where the gearbox technical condition was monitored. When observing vibration signals, we used four tri-axial sensors placed in four selected spots depending on the design arrangement of antifriction bearings. In order to evaluate the measured results, two methods were applied, namely the root mean square gRMS based on the measured acceleration in three axes. The other method used for monitoring the technical condition was the Crest factor. The aim of the experiment was to monitor the technical condition of the mechanical gearbox, mainly its antifriction bearings, therefore the values were measured when the gear was put into fourth position and into overdrive in the auxiliary gearbox. When the gear is put into fourth position, the torque is transmitted from the motor through the main gearbox and the auxiliary gearbox directly. Single gearbox wheels are engaged but not loaded (they do not transmit the torque). At the end of our article we introduced the measured values along with the oral evaluation of the technical condition of the mechanical gearbox and the auxiliary gearbox.

Abstract Background Functional diversity illustrates the range of ecological functions in a community. It allows revealing the appearance of functional redundancy in communities and processes of community assembly. Functional redundancy illustrates the overlap in ecological functions of community members which may be an indicator of community resilience. We evaluated patterns of species richness, functional diversity and functional redundancy on tadpole communities in rainforest streams in Madagascar. This habitat harbours the world's most species-rich stream tadpole communities which are due to their occurrence in primary habitat of particular interest for functional diversity studies. Results Species richness of tadpole communities is largely determined by characteristics of the larval habitat (stream structure), not by adult habitat (forest structure). Species richness is positively correlated with a size-velocity gradient of the streams, i.e. communities follow a classical species-area relationship. While widely observed for other taxa, this is an unusual pattern for anuran larvae which usually is expected to be hump-shaped. Along the species richness gradient, we quantified functional diversity of all communities considering the similarity and dissimilarity of species in 18 traits related to habitat use and foraging. Especially species-rich communities were characterised by an overlap of species function, i.e. by functional redundancy. By comparing the functional diversity of the observed communities with functional diversity of random assemblages, we found no differences at low species richness level, whereas observed species-rich communities have lower functional diversity than respective random assemblages. Conclusions We found functional redundancy being a feature of communities also in primary habitat, what has not been shown before using such a continuous measure. The observed species richness dependent pattern of low functional diversity indicates that communities with low species richness accumulate functional traits randomly, whereas species in species-rich communities are more similar to each other than predicted by random assemblages and therefore exhibit an accumulation of stream-specific functional traits. Beyond a certain species richness level, therefore, stream-specific environmental filters exert influence whereas interspecific competition between species does not influence trait assemblage at any species richness level.

Abstract Background Malagasy tenrecs belong to the Afrotherian clade of placental mammals and comprise three subfamilies divided in eight genera (Tenrecinae: Tenrec, Echinops, Setifer and Hemicentetes; Oryzorictinae: Oryzorictes, Limnogale and Microgale; Geogalinae:Geogale). The diversity of their morphology and incomplete taxon sampling made it difficult until now to resolve phylogenies based on either morphology or molecular data for this group. Therefore, in order to delineate the evolutionary history of this family, phylogenetic and dating analyses were performed on a four nuclear genes dataset (ADRA2B, AR, GHR and vWF) including all Malagasy tenrec genera. Moreover, the influence of both taxon sampling and data partitioning on the accuracy of the estimated ages were assessed. Results Within Afrotheria the vast majority of the nodes received a high support, including the grouping of hyrax with sea cow and the monophyly of both Afroinsectivora (Macroscelidea + Afrosoricida) and Afroinsectiphillia (Tubulidentata + Afroinsectivora). Strongly supported relationships were also recovered among all tenrec genera, allowing us to firmly establish the grouping of Geogale with Oryzorictinae, and to confirm the previously hypothesized nesting of Limnogale within the genus Microgale. The timeline of Malagasy tenrec diversification does not reflect a fast adaptive radiation after the arrival on Madagascar, indicating that morphological specializations have appeared over the whole evolutionary history of the family, and not just in a short period after colonization. In our analysis, age estimates at the root of a clade became older with increased taxon sampling of that clade. Moreover an augmentation of data partitions resulted in older age estimates as well, whereas standard deviations increased when more extreme partition schemes were used. Conclusion Our results provide as yet the best resolved gene tree comprising all Malagasy tenrec genera, and may lead to a revision of tenrec taxonomy. A timeframe of tenrec evolution built on the basis of this solid phylogenetic framework showed that morphological specializations of the tenrecs may have been affected by environmental changes caused by climatic and/or subsequent colonization events. Analyses including various taxon sampling and data partitions allow us to point out some possible pitfalls that may lead to biased results in molecular dating; however, further analyses are needed to corroborate these observations.

Environmental filtering shapes animal communities by preventing the colonization and persistence of certain species in a given habitat. More heterogenous environments are presumed to support a greater number of species and, consequently, increased species diversity, as environmental filters are also likely more heterogenous. Amphibians are especially sensitive to environmental influences due to distinct characteristics like permeable skin and low mobility. By analyzing the species richness and assemblage composition of tadpoles in 132 breeding ponds, we examined how the interplay of environmental variables shapes anuran species assemblages in breeding habitats of the dry forest of Western Madagascar. We found that environmental filtering is prevalent and habitat heterogeneity not only increases larval species richness but also alters species composition between these assemblages. Our study highlights the need for conserving heterogenous habitats to maintain local diversity. Furthermore, we recommend including multivariate modelling approaches to conservation efforts to acknowledge differences between specific habitats and beta diversity.

How environmental factors shape patterns of biotic diversity in tropical ecosystems is an active field of research, but studies examining the possibility of ecological speciation in terrestrial tropical ecosystems are scarce. We use the isolated rainforest herpetofauna on the Montagne d'Ambre (Amber Mountain) massif in northern Madagascar as a model to explore elevational divergence at the level of populations and communities. Based on intensive sampling and DNA barcoding of amphibians and reptiles along a transect ranging from ca. 470-1470 m above sea level (a.s.l.), we assessed a main peak in species richness at an elevation of ca. 1000 m a.s.l. with 41 species. The proportion of local endemics was highest (about 1/3) at elevations >1100 m a.s.l. Two species of chameleons ( Brookesia tuberculata, Calumma linotum ) and two species of frogs ( Mantidactylus bellyi, M. ambony ) studied in depth by newly developed microsatellite markers showed genetic divergence up the slope of the mountain, some quite strong, others very weak, but in each case with genetic breaks between 1100 and 1270 m a.s.l. Genetic clusters were found in transect sections significantly differing in bioclimate and herpetological community composition. A decrease in body size was detected in several species with increasing elevation. The studied rainforest amphibians and reptiles show concordant population genetic differentiation across elevation along with morphological and niche differentiation. Whether this parapatric or microallopatric differentiation will suffice for the completion of speciation is, however, unclear, and available phylogeographic evidence rather suggests that a complex interplay between ecological and allopatric divergence processes is involved in generating the extraordinary species diversity of Madagascar's biota. Our study reveals concordant patterns of diversification among main elevational bands, but suggests that these adaptational processes are only part of the complex of processes leading to species formation, among which geographical isolation is probably also important., Competing Interests: The authors declare no conflicts of interest., (© 2023 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

The distribution of the White-Lipped Frog Amnirana albolabris was long assumed to extend from eastern-central to western African rainforests. However, recent molecular phylogenetic analyses revealed that this taxon likely represents several undescribed species. Because the name-bearing types were collected in Gabon, the distantly related West African populations clearly represent an undescribed species that partly occurs in sympatry with Amnirana fonensis. Based on an integrative taxonomic approach, including molecular, morphological, and acoustic data, we describe the 'albolabris' populations from the Upper Guinean Forest Zone as a new species, and redescribe the morphologically similar A. fonensis on the bases of a larger series of genotyped individuals, including the first known females. We also provide new biological information for A. fonensis, including their advertisement call, habitat, and reproductive data. The new species is sister to A. fonensis and the two species differ by 5.8% in the mitochondrial 16S ribosomal RNA gene. Morphologically, males of the new species are smaller and have relatively smaller humeral glands. Females of the new species seem to have shorter legs than A. fonensis. In comparison to A. fonensis, the advertisement call of the new species has a higher dominant frequency and more pronounced frequency modulation. The two species differ in their distribution and habitat preferences, as revealed by environmental niche modelling. Whereas the new species is restricted to the Upper Guinean forests and thus is a true lowland-rainforest inhabitant, A. fonensis lives predominantly in Guinean montane forests in the forest-savanna mosaic zone.

Phenotypic plasticity may allow ectotherms with complex life histories such as amphibians to cope with climate-driven changes in their environment. Plasticity in thermal tolerance (i.e., shifts of thermal limits via acclimation to higher temperatures) has been proposed as a mechanism to cope with warming and extreme thermal events. However, thermal tolerance and, hence, acclimation capacity, is known to vary with life stage. Using the common frog (Rana temporaria) as a model species, we measured the capacity to adjust lower (CT min ) and upper (CT max ) critical thermal limits at different acclimation temperatures. We calculated the acclimation response ratio as a metric to assess the stage-specific acclimation capacity at each of seven consecutive ontogenetic stages and tested whether acclimation capacity was influenced by body mass and/or age. We further examined how acclimation temperature, body mass, age, and ontogenetic stage influenced CT min and CT max . In the temperate population of R. temporaria that we studied, thermal tolerance and acclimation capacity were affected by the ontogenetic stage. However, acclimation capacity at both thermal limits was well below 100% at all life stages tested. The lowest and highest acclimation capacity in thermal limits was observed in young and late larvae, respectively. The relatively low acclimation capacity of young larvae highlights a clear risk of amphibian populations to ongoing climate change. Ignoring stage-specific differences in thermal physiology may drastically underestimate the climate vulnerability of species, which will hamper successful conservation actions., (© 2022 The Authors. Journal of Experimental Zoology Part A: Ecological Genetics and Integrative Physiology Published by Wiley Periodicals LLC.)

Temperature-induced developmental plasticity could allow amphibian larvae to complete metamorphosis successfully despite new thermal challenges and increased desiccation risk due to climate change. Here we investigated how the capacity for temperature-induced developmental plasticity varies with latitude and whether population-specific biogeographic background accounts for the different degree of plastic responses to temperature. We carried out a combined analysis based on the data from 150 studies (93 articles) performed on 64 amphibian species. We collected empirical data for age and size at metamorphosis in amphibian larvae acclimated to different temperatures during development and found that all larvae from all populations in these studies revealed a change in metamorphic traits with a given change in temperature and thus, were able to exhibit temperature-induced developmental plasticity. Age at metamorphosis was more affected by temperature than size at metamorphosis. Age and size at the onset of metamorphosis were generally lowest at warmest temperatures during development. Furthermore, populations from tropical latitudes were less sensitive to a change in developmental temperature compared to populations from higher latitudes. Accordingly, we suggest tropical populations to be the most vulnerable to increasing temperatures during metamorphosis. Our analyses reveal biases with respect to taxonomy, biogeographic distribution of species, and study design. Data from tropical populations are underrepresented and thus, the capacity for developmental plasticity of the most threatened species probably remains poorly understood. Future studies should focus on under-represented regions, most threatened species, and include a broader range of temperatures during development in order to make robust projections on future sensitivity of populations to climate change., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In the processes that give rise to new species, changes first occur at the population level. But with the continuous nature of the divergence process, change in biological properties delimiting the shift from "individuals of divergent populations" towards "individuals of distinct species", as well as abiotic factors driving the change, remain largely ambivalent. Here we study diversification processes at the population level in a semi-aquatic frog, Mantidactylus (Brygoomantis) bellyi, across the diverse vegetation types of Montagne d'Ambre National Park (MANP), Madagascar. Genetic diversity was assessed with seven newly developed microsatellite markers as well as mitochondrial DNA sequences and concordance with patterns of ecological, morphological, and bioacoustic divergence evaluated. We found M. bellyi lacking mitochondrial differentiation within MANP, while microsatellite datasets partitioned them into three highly differentiated, geographically separated subpopulations (with indications for up to five subpopulations). The molecular grouping-primarily clustering individuals by geographic proximity-was coincident with differences in mean depth and width of waters, suggesting a possible role of fluvial characteristics in genetic exchange in this stream-breeding species. Genetic clustering not consistent with differences in call properties, except for dominant call frequencies under the two-subpopulations model. Morphological divergence was mostly consistent with the genetic clustering; subpopulations strongly differed by their snout-vent length, with individuals from high-elevation subpopulations smaller than those from populations below 1000 m above sea level. These results exemplify how mountains and environmental conditions might primarily shape genetic and morphological divergence in frog populations, without strongly affecting their calls., Competing Interests: The authors have declared that no competing interests exist.

Dispersal is a central process in ecology and evolution with far reaching consequences for the dynamics and genetics of spatially structured populations (SSPs). Individuals can adjust their decisions to disperse according to local fitness prospects, resulting in context-dependent dispersal. By determining dispersal rate, distance and direction, these individual-level decisions further modulate the demography, relatedness and genetic structure of SSPs. Here, we examined how context-dependent dispersal influences the dynamics and genetics of a great crested newt (Triturus cristatus) SSP. We collected capture-recapture data of 5564 individuals and genetic data of 950 individuals across an SSP in northern Germany. We added genetic data from six sites outside this SSP to assess genetic structure and gene flow at a regional level. Dispersal rates within the SSP were high but dispersal distances were short. Dispersal was context-dependent: individuals preferentially immigrated into high-quality ponds where breeding probabilities were higher. The studied SSP behaved like a patchy population, where subpopulations at each pond were demographically interdependent. High context-dependent dispersal led to weak but significant spatial genetic structure and relatedness within the SSP. At the regional level, a strong hierarchical genetic structure with very few first-generation migrants as well as low effective dispersal rates suggest the presence of independent demographic units. Overall, our study highlights the importance of habitat quality for driving context-dependent dispersal and therefore demography and genetic structure in SSPs. Limited capacity for long-distance dispersal seems to increase genetic structure within a population and leads to demographic isolation in anthropogenic landscapes., (© 2021 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

Effective conservation actions require knowledge on the sensitivity of species to pollution and other anthropogenic stressors. Many of these stressors are endocrine disruptors (EDs) that can impair the hypothalamus-pituitary-thyroid axis and thus alter thyroid hormone (TH) levels with physiological consequences to wildlife. Due to their specific habitat requirements, amphibians are often sentinels of environmental degradation. We investigated how altered TH levels affected the bioenergetics of growth and development (i.e. age, size, metabolism, cardiac function and energy stores) before, during and after metamorphosis in the European common frog ( Rana temporaria). We also determined how ontogenetic stage affected susceptibility to endocrine disruption and estimated juvenile performance. TH levels significantly affected growth and energetics at all developmental stages. Tadpoles and froglets exposed to high TH levels were significantly younger, smaller and lighter at all stages compared to those in control and low TH groups, indicating increased developmental and reduced growth rates. Across all ontogenetic stages tested, physiological consequences were rapidly observed after exposure to EDs. High TH increased heart rate by an average of 86% and reduced energy stores (fat content) by 33% compared to controls. Effects of exposure were smallest after the completion of metamorphosis. Our results demonstrate that both morphological and physiological traits of the European common frog are strongly impacted by endocrine disruption and that ontogenetic stage modulates the sensitivity of this species to endocrine disruption. Since endocrine disruption during metamorphosis can impair the physiological stress response in later life stages, long-term studies examining carry-over effects will be an important contribution to the conservation physiology of amphibians., (© The Author(s) 2020. Published by Oxford University Press and the Society for Experimental Biology.)

Anuran larvae show phenotypic plasticity in age and size at metamorphosis as a response to temperature variation. The capacity for temperature-induced developmental plasticity is determined by the thermal adaptation of a population. Multiple factors such as physiological responses to changing environmental conditions, however, might influence this capacity as well. In anuran larvae, thyroid hormone (TH) levels control growth and developmental rate and changes in TH status are a well-known stress response to sub-optimal environmental conditions. We investigated how chemically altered TH levels affect the capacity to exhibit temperature-induced developmental plasticity in larvae of the African clawed frog (Xenopus laevis) and the common frog (Rana temporaria). In both species, TH level influenced growth and developmental rate and modified the capacity for temperature-induced developmental plasticity. High TH levels reduced thermal sensitivity of metamorphic traits up to 57% (R. temporaria) and 36% (X. laevis). Rates of growth and development were more plastic in response to temperature in X. laevis (+30%) than in R. temporaria (+6%). Plasticity in rates of growth and development is beneficial to larvae in heterogeneous habitats as it allows a more rapid transition into the juvenile stage where rates of mortality are lower. Therefore, environmental stressors that increase endogenous TH levels and reduce temperature-dependent plasticity may increase risks and the vulnerability of anuran larvae. As TH status also influences metabolism, future studies should investigate whether reductions in physiological plasticity also increases the vulnerability of tadpoles to global change., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Environmental stress induced by natural and anthropogenic processes including climate change may threaten the productivity of species and persistence of populations. Ectotherms can potentially cope with stressful conditions such as extremes in temperature by exhibiting physiological plasticity. Amphibian larvae experiencing stressful environments display altered thyroid hormone (TH) status with potential implications for physiological traits and acclimation capacity. We investigated how developmental temperature (T dev ) and altered TH levels (simulating proximate effects of environmental stress) influence the standard metabolic rate (SMR), body condition (BC), and thermal tolerance in metamorphic and post-metamorphic anuran larvae of the common frog (Rana temporaria) reared at five constant temperatures (14-28 °C). At metamorphosis, larvae that developed at higher temperatures had higher maximum thermal limits but narrower ranges in thermal tolerance. Mean CT max was 37.63 °C ± 0.14 (low TH), 36.49 °C ± 0.31 (control), and 36.43 °C ± 0.68 (high TH) in larvae acclimated to different temperatures. Larvae were able to acclimate to higher T dev by adjusting their thermal tolerance, but not their SMR, and this effect was not impaired by altered TH levels. BC was reduced by 80% (metamorphic) and by 85% (post-metamorphic) at highest T dev . The effect of stressful larval conditions (i.e., different developmental temperatures and, to some extent, altered TH levels) on SMR and particularly on BC at the onset of metamorphosis was carried over to froglets at the end of metamorphic climax. This has far reaching consequences, since body condition at metamorphosis is known to determine metamorphic success and, thus, is indirectly linked to individual fitness in later life stages.

The genus Heterixalus is endemic to Madagascar. It contains eleven species of small to medium-sized tree frogs that typically inhabit open areas like swamps and rice fields. We describe the larval stages of three sympatric species that occur in western Madagascar: H. tricolor, H. carbonei, and H. luteostriatus. Similar to other species of this genus, the tadpoles of these species have a depressed, ovoid body-form and a generalized oral disc. The labial tooth row formula is 1/3(1). Examined phenotypes differed marginally between species. Compared to H. tricolor and H. carbonei, H. luteostriatus showed fewer cusps on the fork-like labial teeth, a lower ventral fin, and a shorter tail. The high morphological resemblance implicates an ecological similarity between species. Highly overlapping niches raise questions on how species co-occur.

Food conditions are changing due to anthropogenic activities and natural sources and thus, many species are exposed to new challenges. Animals might cope with altered quantitative and qualitative composition [i.e. variable protein, nitrogen (N) and energy content] of food by exhibiting trophic and digestive plasticity. We examined experimentally whether tadpoles of the common frog ( Rana temporaria ) exhibit phenotypic plasticity of the oral apparatus and intestinal morphology when raised on a diet of either low (i.e. Spirulina algae) or high protein, N and energy content (i.e. Daphnia pulex ). Whereas intestinal morphology was highly plastic, oral morphology did not respond plastically to different chemical compositions of food. Tadpoles that were fed food with low protein and N content and low-energy density developed significantly longer guts and a larger larval stomachs than tadpoles raised on high protein, N and an energetically dense diet, and developed a different intestinal surface morphology. Body sizes of the treatment groups were similar, indicating that tadpoles fully compensated for low protein, N and energy diet by developing longer intestines. The ability of a species, R. temporaria , to respond plastically to environmental variation indicates that this species might have the potential to cope with new conditions during climate change., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Environmental change exposes wildlife to a wide array of environmental stressors that arise from both anthropogenic and natural sources. Many environmental stressors with the ability to alter endocrine function are known as endocrine disruptors, which may impair the hypothalamus-pituitary-thyroid axis resulting in physiological consequences to wildlife. In this study, we investigated how the alteration of thyroid hormone (TH) levels due to exposure to the environmentally relevant endocrine disruptor sodium perchlorate (SP; inhibitory) and exogenous L-thyroxin (T4; stimulatory) affects metabolic costs and energy allocation during and after metamorphosis in a common amphibian (Rana temporaria). We further tested for possible carry-over effects of endocrine disruption during larval stage on juvenile performance. Energy allocated to development was negatively related to metabolic rate and thus, tadpoles exposed to T4 could allocate 24% less energy to development during metamorphic climax than control animals. Therefore, the energy available for metamorphosis was reduced in tadpoles with increased TH level by exposure to T4. We suggest that differences in metabolic rate caused by altered TH levels during metamorphic climax and energy allocation to maintenance costs might have contributed to a reduced energetic efficiency in tadpoles with high TH levels. Differences in size and energetics persisted beyond the metamorphic boundary and impacted on juvenile performance. Performance differences are mainly related to strong size-effects, as altered TH levels by exposure to T4 and SP significantly affected growth and developmental rate. Nevertheless, we assume that juvenile performance is influenced by a size-independent effect of achieved TH. Energetic efficiency varied between treatments due to differences in size allocation of internal macronutrient stores. Altered TH levels as caused by several environmental stressors lead to persisting effects on metamorphic traits and energetics and, thus, caused carry-over effects on performance of froglets. We demonstrate the mechanisms through which alterations in abiotic and biotic environmental factors can alter phenotypes at metamorphosis and reduce lifetime fitness in these and likely other amphibians., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.)

Miniaturised frogs form a fascinating but poorly understood amphibian ecomorph and have been exceptionally prone to taxonomic underestimation. The subfamily Cophylinae (family Microhylidae), endemic to Madagascar, has a particularly large diversity of miniaturised species which have historically been attributed to the single genus Stumpffia largely based on their small size. Recent phylogenetic work has revealed that several independent lineages of cophyline microhylids evolved towards highly miniaturised body sizes, achieving adult snout-vent lengths under 16 mm. Here, we describe five new species belonging to three clades that independently miniaturised and that are all genetically highly divergent from their relatives: (i) a new genus (Mini gen. nov.) with three new species from southern Madagascar, (ii) one species of Rhombophryne, and (iii) one species of Anodonthyla. Mini mum sp. nov. from Manombo in eastern Madagascar is one of the smallest frogs in the world, reaching an adult body size of 9.7 mm in males and 11.3 mm in females. Mini scule sp. nov. from Sainte Luce in southeastern Madagascar is slightly larger and has maxillary teeth. Mini ature sp. nov. from Andohahela in southeast Madagascar is larger than its congeners but is similar in build. Rhombophryne proportionalis sp. nov. from Tsaratanana in northern Madagascar is unique among Madagascar's miniaturised frogs in being a proportional dwarf, exhibiting far less advanced signs of paedomorphism than other species of similar size. Anodonthyla eximia sp. nov. from Ranomafana in eastern Madagascar is distinctly smaller than any of its congeners and is secondarily terrestrial, providing evidence that miniaturisation and terrestriality may be evolutionarily linked. The evolution of body size in Madagascar's microhylids has been more dynamic than previously understood, and future studies will hopefully shed light on the interplay between ecology and evolution of these remarkably diverse frogs., Competing Interests: The authors have declared that no competing interests exist.

Environmental variation induced by natural and anthropogenic processes including climate change may threaten species by causing environmental stress. Anuran larvae experiencing environmental stress may display altered thyroid hormone (TH) status with potential implications for physiological traits. Therefore, any capacity to adapt to environmental changes through plastic responses provides a key to determining species vulnerability to environmental variation. We investigated whether developmental temperature ( T dev ), altered TH levels and whether the interactive effect of both affect standard metabolic rate (SMR), body condition (BC), survival and thermal tolerance in larvae of the African clawed frog ( Xenopus laevis ) reared at five temperatures with experimentally altered TH levels. At metamorphosis, SMR, BC and survival were significantly affected by T dev , TH status and their interaction with the latter often intensified impacts. Larvae developing at warmer temperatures exhibited significantly higher SMRs and BC was reduced at warm T dev and high TH levels suggesting decreased ability to acclimate to variation in temperature. Accordingly, tadpoles that developed at warm temperatures had higher maximum thermal limits but more narrow thermal tolerance windows. High and low TH levels decreased and increased upper thermal limits, respectively. Thus, when experiencing both warmer temperatures and environmental stress, larvae may be less able to compensate for changes in T dev . Our results demonstrate that physiological traits in larvae of X. laevis are strongly affected by increased TH levels and warmer temperatures. Altered TH levels and increasing T dev due to global change may result in a reduced capacity for physiological plasticity. This has far reaching consequences since the energetic requirement at the onset of metamorphosis is known to determine metamorphic success and thus, is indirectly linked to individual fitness in later life stages.

Chemical, physical and biological environmental stressors may affect the endocrine system, such as the thyroid hormone (TH) axis in larval amphibians with consequences for energy partitioning among development, growth and metabolism. We studied the effects of two TH level affecting compounds, exogenous l-thyroxine (T 4 ) and sodium perchlorate (SP), on various measures of development and body condition in larvae of the African clawed frog (Xenopus laevis). We calculated the scaled mass index, hepatosomatic index and relative tail muscle mass as body condition indices to estimate fitness. Altered TH levels significantly altered the growth, development, survival and body condition in metamorphic larvae in different directions. While exogeno us T 4 reduced growth and accelerated development, SP treatment increased growth but slowed down development. Altered TH levels improved body conditions in both treatments and particularly in larvae of the SP treatment but to the detriment of lower survival rates in both TH level altering treatments. The hepatosomatic index was negatively affected by exogenous T 4 , but not by SP treatment indicating a lower lipid reserve in the liver in larvae of T 4 treatment. These altered TH levels as caused by several environmental stressors may have an influence on individual fitness across life, as body condition at the onset of metamorphosis determines metamorphic and juvenile survival. Further research is needed to determine synergetic effects of environmental stressors on TH levels and its effects on physiological traits such as metabolic rate., (© 2018 John Wiley & Sons, Ltd.)

Anurans exhibit plasticity in the timing of metamorphosis and tadpoles show phenotypic plasticity in age and size at metamorphosis as a response to temperature variation. This developmental plasticity to changing thermal conditions is expected to be a primary factor that dictates the vulnerability of amphibians to increasing ambient temperatures such as are predicted in climate change scenarios. We analyzed the patterns of thermal effects on size and age at metamorphosis to investigate whether the intraspecific "temperature-size rule" is applicable over a broad range of anuran species by carrying out a combined analysis based on the data from 25 studies performed on 18 anuran species. Furthermore, we tested whether the thermal background of respective populations impacts the capacity for a plastic response in metamorphic traits. We could confirm this pattern for across-population comparisons. All included populations developed faster and 75% were smaller at the onset of metamorphosis when developmental temperatures were warmer, but the sensitivity of growth and developmental rate to a given temperature change was different. We found that the thermal background of a population influences the sensitivity of metamorphic traits and thus, the capacity for a plastic response in growth and developmental rate. Warm adapted populations were less sensitive to temperature variation indicating a reduced capacity for developmental plasticity and therefore, those species may be more vulnerable to the impacts of climate change. Future studies should include a broader range of rearing temperatures and temperature fluctuations to determine full knowledge of the capacity for developmental plasticity within a species-specific thermal window., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Animal-associated microbial communities can play major roles in the physiology, development, ecology, and evolution of their hosts, but the study of their diversity has yet focused on a limited number of host species. In this study, we used high-throughput sequencing of partial sequences of the bacterial 16S rRNA gene to assess the diversity of the gut-inhabiting bacterial communities of 212 specimens of tropical anuran amphibians from Brazil and Madagascar. The core gut-associated bacterial communities among tadpoles from two different continents strongly overlapped, with eight highly represented operational taxonomic units (OTUs) in common. In contrast, the core communities of adults and tadpoles from Brazil were less similar with only one shared OTU. This suggests a community turnover at metamorphosis. Bacterial diversity was higher in tadpoles compared to adults. Distinct differences in composition and diversity occurred among gut bacterial communities of conspecific tadpoles from different water bodies and after experimental fasting for 8 days, demonstrating the influence of both environmental factors and food on the community structure. Communities from syntopic tadpoles clustered by host species both in Madagascar and Brazil, and the Malagasy tadpoles also had species-specific isotope signatures. We recommend future studies to analyze the turnover of anuran gut bacterial communities at metamorphosis, compare the tadpole core communities with those of other aquatic organisms, and assess the possible function of the gut microbiota as a reservoir for protective bacteria on the amphibian skin.

Assemblages that are exposed to recurring temporal environmental changes can show changes in their ecological properties. These can be expressed by differences in diversity and assembly rules. Both can be identified using two measures of diversity: functional (FD) and phylogenetic diversity (PD). Frog communities are understudied in this regard, especially during the tadpole life stage. We utilised tadpole assemblages from Madagascan rainforest streams to test predictions of seasonal changes on diversity and assemblage composition and on diversity measures. From the warm-wet to the cool-dry season, species richness (SR) of tadpole assemblages decreased. Also FD and PD decreased, but FD less and PD more than expected by chance. During the dry season, tadpole assemblages were characterised by functional redundancy (among assemblages-with increasing SR), high FD (compared to a null model), and low PD (phylogenetic clustering; compared to a null model). Although mutually contradictory at first glance, these results indicate competition as tadpole community assembly driving force. This is true during the limiting cool-dry season but not during the more suitable warm-wet season. We thereby show that assembly rules can strongly depend on season, that comparing FD and PD can reveal such forces, that FD and PD are not interchangeable, and that conclusions on assembly rules based on FD alone are critical.

We describe a new species of small-sized frogs from degraded rainforest patches in the southern central east of Madagascar. Stumpffia miery sp. nov. has a snout-vent length of 13-15 mm and can be distinguished from all other nominal species of Stumpffia by its body size and absence of toe reduction combined with length reduction of fingers I, II and IV in external view. The advertisement call is a single tonal chirping note that ranges in duration between 51-88 ms and is emitted after relatively regular inter-note intervals (duration of 2679-4247 ms, call repetition rate 0.3/sec, frequency range 7700-8300 Hz, dominant frequency 7751-8225 Hz). Its type locality is the Ambolo forest fragment close to Ranomafana village in southeastern Madagascar. Molecular data from DNA sequences of one mitochondrial and one nuclear gene indicate a high divergence from all nominal species of Stumpffia, suggesting that it represents a strongly differentiated independent evolutionary unit. Stumnpffia miery sp. nov. is apparently able to tolerate some degree of habitat degradation and therefore is probably not threatened with extinction.

This study provides detailed morphological descriptions of previously unknown tadpoles of the treefrog genus Boophis Tschudi and analyses of habitat preferences of several of these tadpoles in Ranomafana National Park. A total of twenty-two tadpoles determined via DNA barcoding are characterized morphologically herein, fourteen of them for the first time. Twelve of these tadpoles belong to taxonomically undescribed candidate species which in several cases are so far only known from their larval stages. Our data show that the larvae of some of these candidate species occur syntopically yet maintaining a clearly correlated genetic and morphological identity, suggesting that they indeed are true biological and evolutionary species. Tadpoles considered to belong to the "adherent" ecomorphological guild inhabit fast-running waters and their oral disc is commonly to continuously attached to the rocky substrate, supposedly to keep their position in the water current. Some of these species are characterized by the presence of a dorsal gap of papillae and the absence of an upper jaw sheath. This guild includes the tadpoles of the Boophis albipuncatus group (Boophis ankaratra, Boophis schuboeae, Boophis albipunctatus, Boophis sibilans, Boophis luciae), and of the Boophis mandraka group (Boophis sambirano and six candidate species related to this species and to Boophis mandraka). Tadpoles considered belonging to the "suctorial" guild inhabit fast-running waters where they use frequently their oral disc to attach to the substrate. They have an enlarged oral disc without any dorsal gap, including two nominal species (Boophis marojezensis, Boophis vittatus), and five candidate species related to Boophis marojezensis. An ecological analysis of the tadpoles of Boophis luciae, Boophis schuboeae and Boophis marojezensis [Ca51 JQ518198] from Ranomafana National Park did not provide evidence for a clear preference of these tadpoles to the fast flowing microhabitat sections of the stream, although the tadpoles discussed in this study are typically caught in this habitat.

Acoustic communication is widespread among adult stages of terrestrial animals and fish and has also been observed in insect larvae. We report underwater acoustic communication in the larvae of a frog, Gephyromantis azzurrae, from Isalo, a sandstone massif in western Madagascar. According to our field data, these tadpoles live in streams and prefer habitats characterized by comparatively low temperatures, shallow water depth, and a relatively fast current. Feeding experiments indicated that the tadpoles are carnivorous and macrophagous. They consumed insect larvae and, to a lesser extent, small shrimps, and conspecific as well as heterospecific tadpoles. Calls of these tadpoles consisted either of single click notes or of irregular series of various clicks. Some complex calls have a pulsed structure with three to nine indistinct energy pulses. Production of the pulses coincided with rapid closure of the jaw sheaths and often with an upward movement of the body. Calls were emitted while attacking prey and occurred significantly more often when attacking conspecifics. Tadpoles that had not been fed for some time emitted sounds more frequently than those that had been regularly fed. The spectral frequency of the calls differed in tadpole groups of different size and was higher in groups of smaller tadpoles, suggesting that spectral frequency carries some information about tadpole size which might be important during competitive feeding to assess size and strength of competitors. This report differs from those for the larvae of South American horned frogs, Ceratophrys ornata. These are the only other tadpoles for which sound production has reliably been reported but the calls of Ceratophrys tadpoles occur mainly in a defensive context.

Background: Functional diversity illustrates the range of ecological functions in a community. It allows revealing the appearance of functional redundancy in communities and processes of community assembly. Functional redundancy illustrates the overlap in ecological functions of community members which may be an indicator of community resilience. We evaluated patterns of species richness, functional diversity and functional redundancy on tadpole communities in rainforest streams in Madagascar. This habitat harbours the world's most species-rich stream tadpole communities which are due to their occurrence in primary habitat of particular interest for functional diversity studies., Results: Species richness of tadpole communities is largely determined by characteristics of the larval habitat (stream structure), not by adult habitat (forest structure). Species richness is positively correlated with a size-velocity gradient of the streams, i.e. communities follow a classical species-area relationship. While widely observed for other taxa, this is an unusual pattern for anuran larvae which usually is expected to be hump-shaped. Along the species richness gradient, we quantified functional diversity of all communities considering the similarity and dissimilarity of species in 18 traits related to habitat use and foraging. Especially species-rich communities were characterised by an overlap of species function, i.e. by functional redundancy. By comparing the functional diversity of the observed communities with functional diversity of random assemblages, we found no differences at low species richness level, whereas observed species-rich communities have lower functional diversity than respective random assemblages., Conclusions: We found functional redundancy being a feature of communities also in primary habitat, what has not been shown before using such a continuous measure. The observed species richness dependent pattern of low functional diversity indicates that communities with low species richness accumulate functional traits randomly, whereas species in species-rich communities are more similar to each other than predicted by random assemblages and therefore exhibit an accumulation of stream-specific functional traits. Beyond a certain species richness level, therefore, stream-specific environmental filters exert influence whereas interspecific competition between species does not influence trait assemblage at any species richness level.