Spatial network structure and amphibian persistence in stochastic environments

In the past few years, the framework of complex networks has provided new insight into the organization and function of biological systems. However, in spite of its potential, spatial ecology has not yet fully incorporated tools and concepts from network theory. In the present study, we identify a large spatial network of temporary ponds, which are used as breeding sites by several amphibian species. We investigate how the structural properties of the spatial network change as a function of the amphibian dispersal distance and the hydric conditions. Our measures of network topology suggest that the observed spatial structure of ponds is robust to drought (compared with similar random structures), allowing the movement of amphibians to and between flooded ponds, and hence, increasing the probability of reproduction even in dry seasons.     

Chytrid fungus parasitizing the wild amphibian Leptodactylus ocellatus (Anura: Leptodactylidae) in Argentina

The present contribution is the first report of parasitosis by a chytrid fungus in wild anuran amphibians in Argentina, as well as the first case of amphibian mortality documented to date in Argentina. We report the presence of the chytrid fungus in dead adult Leptodactylus ocellatus. It has been suggested that chytridiomycosis is the main cause of death in several amphibian populations worldwide. Our study demonstrates that chytridiomycosis afflicts L. ocellatus, a common widespread amphibian species, and is the first report of chytridiomycosis in the Argentinian lowlands. The occurrence at this latitude would indicate an extended distribution of this fungus in wildlife populations. It is also the first report of amphibian mortality due to chytrid fungus in our country. It is noteworthy that the site of collection is situated very close to sea level in a temperate climate zone and that this represents the southern most record for South American wild amphibians.     

My perpetual cycle: from student to researcher to teacher to student ..

This contribution stems from the personal experience of the author regarding how he became acquainted with embryology and how he finally entered the field of developmental biology. It reports his feelings as a student of the Histology and Embryology course as it was taught in the late 1970s, and his present efforts in teaching developmental biology to university students. In the Developmental Biology course at Pisa University today, students are taught the tissue, molecular and genetic mechanisms that regulate development of several model systems. Drosophila is introduced at the beginning, because of the great knowledge that it has brought to the unraveling of the molecular aspects of development and because it allows several basic concepts to be introduced, and vertebrate systems follow. Other topics include the classic experiments on amphibian systems, which are explained in the light of recent molecular advances, as well as the genetically more versatile vertebrate systems such as the mouse.     

Wetlands classification for amphibian conservation in Mediterranean landscapes

Habitat fragmentation and alteration have been described as the major factors relating to the decline observed in several amphibian species. In Spain, these processes are mainly associated with the large agricultural land area, the widespread use of toxic substances, and the agricultural practices used. All these aggressions affect wetlands and their watersheds, which are essential aquatic systems for many amphibian species. In an effort to discover where conservation and specific action plans could be more necessary, this study relates the amphibian richness in 28 representative Mediterranean wetlands to their characteristics and the activities carried out on their watershed. The required information considers diverse wetland variables. Our results show that watershed habitat heterogeneity is significantly positively correlated with amphibian richness, while the percentage of the watershed area devoted to olive cultivation is negatively correlated with it. The results allow us to classify wetlands according to their importance for amphibian conservation. Our results also show that 80% of the studied wetlands need specific action plans. Especially, more sustainable cultivation methods are required considering drastic changes in current agricultural practices. Finally, our data provides a baseline to understand future alterations due to agricultural practices.     

The global amphibian trade flows through Europe: the need for enforcing and improving legislation

The global amphibian trade is suspected to have brought several species to the brink of extinction, and has led to the spread of amphibian pathogens. Moreover, international trade is not regulated for ~98 % of species. Here we outline patterns and complexity underlying global amphibian trade, highlighting some loopholes that need to be addressed, focusing on the European Union. In spite of being one of the leading amphibian importers, the EU’s current legislation is insufficient to prevent overharvesting of those species in demand or the introduction and/or spread of amphibian pathogens into captive and wild populations. We suggest steps to improve the policy (implementation and enforcement) framework, including (i) an identifier specifically for amphibians in the World Customs Organisation’s harmonised system, (ii) Parties to CITES should strive to include more species in the CITES appendices, and (iii) restriction or suspension of trade of threatened species, restricted-range species, and species protected in their country of origin. Commercial trade should not put survival of amphibian species further at risk.     

Darwin,the amphibians,and the natural selection

A critical review of Darwin's publications shows that he did not dissert much about amphibians, in comparison with the other tetrapods. However, in “A Naturalist's Voyage round the World”, Darwin described for the first time several amphibian species and was surprised by their peculiar way of life, terrestrial or euryhaline. These amphibian observations around the world led Darwin to discuss evolutionnary notions, like developmental heterochronies or evolving convergences, and later to illustrate his famous natural selection theory. This is confirmed, for example, by the publication of “On the Origin of Species” where Darwin ironically questioned creation theory, trying to explain the absence of amphibians on oceanic islands. Lamarck also considered amphibians as relevant material to illustrate his theory of acquired character heredity. These historical uses of lissamphibians as evolutionary models have been mostly realized before any amphibian fossil discovery, i.e. out of a palaeontological context.     

Widespread infection of the Eastern red-spotted newt (Notophthalmus viridescens) by a new species of Amphibiocystidium, a genus of fungus-like mesomycetozoan parasites not previously reported in North America

Given the worldwide decline of amphibian populations due to emerging infectious diseases, it is imperative that we identify and address the causative agents. Many of the pathogens recently implicated in amphibian mortality and morbidity have been fungal or members of a poorly understood group of fungus-like protists, the mesomycetozoans. One mesomycetozoan, Amphibiocystidium ranae, is known to infect several European amphibian species and was associated with a recent decline of frogs in Italy. Here we present the first report of an Amphibiocystidium sp. in a North American amphibian, the Eastern red-spotted newt (Notophthalmus viridescens), and characterize it as the new species A. viridescens in the order Dermocystida based on morphological, geographical and phylogenetic evidence. We also describe the widespread and seasonal distribution of this parasite in red-spotted newt populations and provide evidence of mortality due to infection.     

湖南壶瓶山国家级自然保护区两栖 爬行动物多样性的时空格局

本研究利用2013至2016年湖南壶瓶山国家级自然保护区巡护监测数据,评估壶瓶山保护区两栖爬行动物多样性水平。壶瓶山保护区在8个监测站点布设了25条两栖爬行动物调查样线,进行常规监测,其中,日间样线14条,夜间样线11条,日间样线平均长度4 km,夜间样线平均长度0.75 km。每个月,每条样线平均调查3次。4年共调查到62种两栖爬行动物,占湖南省两栖爬行动物总种数的38.04%。逐月计算各监测站点的alpha多样性指数和整个保护区的beta多样性指数,按季节比较不同站点间物种组成的Hellinger距离。结果显示,保护区不同位置间、不同的监测站点间以及年际间的两栖爬行动物多样性没有显著差异,提示保护区环境具有相对稳定性。但是,个别位于实验区或邻近实验区的站点,物种组成有着剧烈的波动,表明人为活动影响了两栖爬行动物的分布。另外,保护区两栖爬行动物的物种组成与多样性有着显著的季节差异,这与两栖爬行动物的生物学特性有关。以上结果说明,壶瓶山保护区两栖爬行动物的多样性结构具有稳定性和敏感性的特点,可以考虑用作保护区生物多样性水平的监测指标。     

Gastrulation in amphibian embryos, regarded as a succession of biomechanical feedback events

Gastrulation in amphibian embryos is a composition of several differently located morphogenetic movements which are perfectly coordinated with each other both in space and time. We hypothesize that this coordination is mediated by biomechanical interactions between different parts of a gastrulating embryo based upon the tendency of each part to hyper-restore the value of its mechanical stress. The entire process of gastrulation in amphibian embryos is considered as a chain of these mutually coupled reactions, which are largely dependent upon the geometry of a given embryo part. We divide gastrulation into several partly overlapped steps, give a theoretical interpretation for each of them, formulate the experiments for testing our interpretation and describe the experimental results which confirm our point of view. Among the predicted experimental results are: inhibition of radial cell intercalation by relaxation of tensile stresses at the blastula stage; inversion of convergent intercalation movements by relaxation of circumferential stresses at the early gastrula stage; stress-promoted reorientation of axial rudiments, and others. We also show that gastrulation is going on under a more or less constant average value of tensile stresses which may play a role as rate-limiting factors. A macro-morphological biomechanical approach developed in this paper is regarded as complementary to exploring the molecular machinery of gastrulation.     

Amphibian declines: future directions

Abstract. The amphibian decline problem is complex, and there is no easy solution. I highlight four major areas of future research that should increase our ability to detect declines, elucidate their underlying mechanisms, and advance our capacity to manage and conserve amphibian populations. First, a statistically sensitive monitoring approach is necessary to determine the distribution and abundance of amphibian populations, to assess whether they are declining, and to quantify the extent of declines. Most amphibian populations characteristically fluctuate, detection probabilities may be low for many species and populations tend to decline in numbers between years more often than they increase. These traits make establishing monitoring programmes difficult and distinguishing declines from natural fluctuations challenging. It is thus necessary to determine the best monitoring techniques based on their statistical power and to use appropriate statistical methods for detecting population trends. Secondly, although amphibian population studies occur most commonly at single or few breeding sites, research should occur often at the landscape level, and conservation efforts should focus on suitable habitat (whether or not it is occupied) and dispersal capabilities of species. Metapopulation dynamics are probably important for many species, but we must be cautious how we define metapopulations. That is, the term ‘metapopulation’ is currently used to define a wide range of demographic situations in amphibian populations, each with different management implications. Thirdly, recent advances in molecular genetic techniques make it possible to infer demographic events such as effects of recent fragmentation, bottlenecks or hybridization. Molecular techniques can be used in conjunction with census surveys to bolster knowledge about demographic processes such as declines. Alternatively, in the absence of long‐term census data, molecular data can be used to infer population trends. New genomic approaches may make estimating adaptive genetic variation more feasible. Fourthly, multi‐factorial studies are needed to disentangle the complexity of the several putative causes that probably interact to cause amphibian declines. Recent studies demonstrate the value of a multi‐factorial approach, and more work is needed to elucidate the synergistic effects of multiple environmental factors affecting amphibian populations simultaneously worldwide.     

环境DNA在两栖动物监测中的应用研究进展

两栖动物是我国受威胁程度最高的动物类群,加强两栖动物资源调查和多样性监测,是开展两栖动物保护和濒危物种拯救行动的关键性基础工作。传统的两栖动物监测主要以形态学和声学为基础,耗时费力,且难以发现一些隐蔽性较强的稀有物种。基于环境DNA(environmental DNA, eDNA)的调查方法以其快速、灵敏、高效、无创等独特优势,为两栖动物多样性监测及保护提供了新的工具。综述了eDNA在两栖动物多样性监测、外来入侵和珍稀濒危物种调查、物种丰度或生物量估测等研究领域的应用进展,分析了两栖动物eDNA产生、扩散、迁移和降解的动态变化特征及其关键影响因子,探讨了eDNA应用于两栖动物监测研究的局限性并提出了优化建议,同时对未来的研究方向进行了展望,以充分挖掘eDNA在两栖动物监测中的应用潜力,为两栖动物多样性保护和管理提供新的思路。     

Amphibian species detection in water reservoirs using artificial neural networks for ecology-friendly city planning

Planning cities regardless of natural resources has led to the existence of threats for several species that occupy an important place in local ecosystems. Constructing buildings, bridges, or motorways over water reservoirs is perilous for resident amphibians, which belong to one of the most fragile animal groups. An automated system that reliably identifies whether any amphibian species live in a water reservoir before construction in that region will help building inspectors during city planning and will ensure that citizens can enjoy the opportunities provided by nature. In this paper, an intelligent amphibian species detection system is developed using features extracted from GIS and satellite images. The presence of seven amphibian species is detected using an artificial neural network (ANN). Depending on the spatial features of the region, the proposed cascade-forward backpropagation neural network (CFBNN) model, a special type of ANN, determines which amphibian species live there to enable ecological precautions to be taken before construction starts and to preserve the biodiversity for the future health of the community. The results clearly demonstrate that the proposed approach significantly outperforms recently suggested strategies with more than 15% improvement on average. The performance of the system shows that the proposed detection approach can be a useful tool for smart and ecological city planning.     

Amphibian teeth: current knowledge, unanswered questions, and some directions for future research

Elucidation of the mechanisms controlling early development and organogenesis is currently progressing in several model species and a new field of research, evolutionary developmental biology, which integrates developmental and comparative approaches, has emerged. Although the expression pattern of many genes during tooth development in mammals is known, data on other lineages are virtually non-existent. Comparison of tooth development, and particularly of gene expression (and function) during tooth morphogenesis and differentiation, in representative species of various vertebrate lineages is a prerequisite to understand what makes one tooth different from another. Amphibians appear to be good candidates for such research for several reasons: tooth structure is similar to that in mammals, teeth are renewed continuously during life (=polyphyodonty), some species are easy to breed in the laboratory, and a large amount of morphological data are already available on diverse aspects of tooth biology in various species. The aim of this review is to evaluate current knowledge on amphibian teeth, principally concerning tooth development and replacement (including resorption), and changes in morphology and structure during ontogeny and metamorphosis. Throughout this review we highlight important questions which remain to be answered and that could be addressed using comparative morphological studies and molecular techniques. We illustrate several aspects of amphibian tooth biology using data obtained for the caudate Pleurodeles waltl. This salamander has been used extensively in experimental embryology research during the past century and appears to be one of the most favourable amphibian species to use as a model in studies of tooth development.     

A comparative analysis of global, national, and state red lists for threatened amphibians in Brazil

The knowledge about the conservation status of species is an important data for conservation biology. Therefore, threatened species lists are a powerful tool for conservation planning and prioritization. Our objective is to compare the global, the national and state red lists of amphibians in Brazil. Threatened species were categorized according to their listing in one or several of these lists. We analyzed for true inconsistencies across lists in order to evaluate practical consequences of such incongruences on amphibian conservation in Brazil. We recorded a total of 61 threatened amphibian species in Brazil (across all red lists). Only one species, Phrynomedusa fimbriata, was listed as Extinct (both in IUCN, Brazil and S?o Paulo lists). A total of eleven endemic species are listed as threatened by the global red list, but do not appear in Brazil’s national red list, which represent an inconsistence among these lists. Besides that, the threat category of Thoropa lutzi and Thoropa petropolitana, two endemic species, differ among both lists, which also represents a problem between both lists. These mismatches may be due to several reasons such as different interpretation of the criteria; different methodologies used; different data availability on species; differences in the dates of assessments processes; the assessors’ attitudes to uncertainty; outdated red lists. Harmonization among red lists permits a better picture of threatened amphibian diversity across scales and to develop global, national and state plans to complement conservation actions in order to maximize the chance of success of these initiatives.     

Apical material extracted from amphibian urinary bladder epithelium by enzymes and detergent treatment

The apical plasma membrane of epithelial cells of frog and toad urinary bladder is subject to large modifications during the induction of water permeability by the antidiuretic hormone. A better characterization of the apical membrane is necessary for a clear understanding of the mechanisms of hormone action. Towards this end, apical material was extracted by enzymatic treatment and by incubation with detergent. Proteolytic enzyme alone had little effect under our conditions. A pretreatment with several glycosidases (alpha-mannosidase or endo-beta-N-acetylglucosaminidase H) increased the hydrolytic action of papain, elastase, proteinase K or Staphylococcus aureus V8 protease and allowed the detection of a major 76 kD in SDS gel electrophoresis. The n-octyl-beta-D-glucopyranoside (0.2%) led to the extraction after 150 mn of 1 to 5 micrograms proteins per cm2 of amphibian urinary bladder apical surface. The extracted proteins migrated as several bands on SDS gels. One of them probably corresponds to the 76 kD fragment obtained after proteolysis. The absence of alteration of the water permeability after extraction and the good preservation of the ultrastructure are evidence for the localisation of the 76 kD at the apical membrane surface. This protein may be the best candidate as antigen to raise antibodies against the apical surface of amphibian urinary bladder epithelial cells.     

Chromosome banding in Amphibia

Fixed metaphase chromosomes of several species of Amphibia were treated with various restriction endonucleases and subsequently stained with Giemsa. Metaphases of man and chicken were examined in parallel under the same experimental conditions for comparison. The restriction enzymes always induce subsets of the C-banding patterns present in the amphibian karyotypes. The heterochromatic regions can be either resistant or sensitive to the restriction enzyme. The modified C-banding patterns revealed by different restriction endonucleases in the karyotype of the same species can be either extremely dissimilar or almost completely congruent. Correspondingly, the action of the same restriction enzyme on the karyotypes of different species may vary greatly. There is only rarely a correlation between the type of C-banding patterns produced by different restriction endonucleases and their specific base pair recognition sequences. In contrast to mammalian and avian chromosomes, restriction enzymes induce no multiple G-banding patterns in amphibian chromosomes. This is attributed to the difference in organization of the DNA in the genomes of poikilothermic vertebrates. The possible mechanisms of restriction endonuclease banding and the various uses of this technique for amphibian chromosomes are discussed.     

Amphibian organ remodeling during metamorphosis: insight into thyroid hormone-induced apoptosis

During amphibian metamorphosis, the animal body dramatically remodels to adapt from the aquatic to the terrestrial life. Cell death of larval organs/tissues occurs massively in balance with proliferation of adult organs/tissues, to ensure survival of the individuals. Thus, amphibian metamorphosis provides a unique and valuable opportunity to study regulatory mechanisms of cell death. The advantage of this animal model is the absolute dependence of amphibian metamorphosis on thyroid hormone (TH). Since the 1990s, a number of TH response genes have been identified in several organs of Xenopus laevis tadpoles such as the tail and the intestine by subtractive hybridization and more recently by cDNA microarrays. Their expression and functional analyses, which are still ongoing, have shed light on molecular mechanisms of TH‐induced cell death during amphibian metamorphosis. In this review, I survey the recent progress of research in this field, focusing on the X. laevis intestine where apoptotic process is well characterized at the cellular level and can be easily manipulated in vitro. A growing body of evidence indicates that apoptosis during the intestinal remodeling occurs not only via a cell‐autonomous pathway but also via cell–cell and/or cell–extracellular matrix (ECM) interactions. Especially, stromelysin‐3, a matrix metalloproteinase, has been shown to alter cell–ECM interactions by cleaving a laminin receptor and induce apoptosis in the larval intestinal epithelium. Here, I emphasize the importance of TH‐induced multiple apoptotic pathways for massive and well‐organized apoptosis in the amphibian organs and discuss their conservation in the mammalian organs.     

Global amphibian declines: sorting the hypotheses

Abstract. Reports of malformed amphibians and global amphibian declines have led to public concern, particularly because amphibians are thought to be indicator species of overall environmental health. The topic also draws scientific attention because there is no obvious, simple answer to the question of what is causing amphibian declines? Complex interactions of several anthropogenic factors are probably at work, and understanding amphibian declines may thus serve as a model for understanding species declines in general. While we have fewer answers than we would like, there are six leading hypotheses that we sort into two classes. For class I hypotheses, alien species, over‐exploitation and land use change, we have a good understanding of the ecological mechanisms underlying declines; these causes have affected amphibian populations negatively for more than a century. However, the question remains as to whether the magnitude of these negative effects increased in the 1980s, as scientists began to notice a global decline of amphibians. Further, remedies for these problems are not simple. For class II hypotheses, global change (including UV radiation and global climate change), contaminants and emerging infectious diseases we have a poor, but improving understanding of how each might cause declines. Class II factors involve complex and subtle mechanistic underpinnings, with probable interactions among multiple ecological and evolutionary variables. They may also interact with class I hypotheses. Suspected mechanisms associated with class II hypotheses are relatively recent, dating from at least the middle of the 20th century. Did these causes act independently or in concert with pre‐existing negative forces of class I hypotheses to increase the rate of amphibian declines to a level that drew global attention? We need more studies that connect the suspected mechanisms underlying both classes of hypotheses with quantitative changes in amphibian population sizes and species numbers. An important step forward in this task is clarifying the hypotheses and conditions under which the various causes operate alone or together.     

A criticism of catastrophe modelling of the differentiative process in amphibian development

A criticism of Zeeman's (1975) general theory of developmental organization, with special reference to amphibian embryogenesis, and based on a mathematical theorem using catastrophe theory to model the process of differentiation, leads to the conclusion that the model itself is not a rigorous scientific theory. Catastrophe theory in itself offers no new insight, merely a modelling of concepts of continuous organization, of process. These concepts are of considerable interest, but the claim that the theorem proves the necessity for primary waves in development is spurious.