Histology and microscopic anatomy of the thyroid gland during the larval development of Pseudis platensis (Anura,Hylidae)

Several hormones regulate anuran larval development, most notably thyroid hormones (THs). In anurans, metamorphosis fails when the thyroid gland is absent or inactivated, resulting in giant tadpoles. Larval gigantism occurs naturally in neotropical frogs of the genus Pseudis as a result of a prolonged larval period. Its thyroid function is poorly investigated and the focus of this study. We describe qualitative and quantitative variations in larval development for field-captured specimens of Pseudis platensis and compare those to the development of two sympatric species, Phyllomedusa sauvagii and Pithecopus azureus, which have small tadpoles and a shorter larval period. We describe morphological changes in the thyroid glands of larval and adult specimens. In contrast to other species with similar ecological requirements, P. platensis exhibits distinct glandular activity. During premetamorphosis, there was little or no thyroid activity, a period in which the tadpole reached 70% of its maximum size. Development and degree of activity of the thyroid gland determine the duration of the early stages of the larval period. Thyroid gland histology in tadpoles appears to correlate with the TH activity, and in turn with the diversity in anuran life history transitions.     

Ancient ontogenies: larval development of the Lower Cretaceous anuran Shomronella jordanica (Amphibia: Pipoidea)

The Lower Cretaceous anuran Shomronella jordanica (Pipoidea) is represented by an assemblage of nearly 300 tadpoles of different ages. The size of the assemblage allows a reconstruction of the larval ontogeny of this species. We describe the ossification sequence and growth rates of S. jordanica and present reconstructions of tadpoles at different stages. The ontogeny of the species seems to be similar to that of extant pipids in many aspects. Larvae are similar in shape and size to those of Xenopus laevis but lack the anterior barbels that are typical for all extant pipids. The ossification sequence is closer to that of pipids than to that of other anurans. We present evidence that suggests that this species was terrestrial as an adult, and we discuss the implications of these data for the evolution of anuran development.     

Developmental changes in interrenal responsiveness in anuran amphibians

Basal activity of the hypothalamo-pituitary-interrenal (HPI)axis changes over development in larval amphibians, but developmentof the responsiveness of this axis to an external stressor hasnot been studied. We compared developmental changes in whole-bodycorticosterone content of two anuran amphibian species, Ranapipiens (family Ranidae) and Xenopus laevis (family Pipidae).We also examined developmental changes in the responsivenessof the HPI axis by subjecting tadpoles of different developmentalstages to a laboratory shaking/confinement stress and to ACTHinjection. We measured whole-body corticosterone content asan indicator of the activity of the HPI axis. Whole-body corticosteronecontent of R. pipiens remained low during premetamorphosis andprometamorphosis but increased dramatically at metamorphic climaxand remained elevated in juvenile frogs. By contrast, whole-bodycorticosterone content of X. laevis was highest during premetamorphosis,declined at the onset of prometamorphosis, increased at metamorphicclimax and remained at climax levels in juvenile frogs. Premetamorphicand prometamorphic tadpoles of both species showed strong corticosteroneresponses to both shaking stress and ACTH injection. The magnitudeand pattern of response differed among developmental stages,with premetamorphic tadpoles of both species showing greaterresponsiveness to stress and ACTH. Our results show that interrenalresponsiveness is developed in premetamorphic tadpoles, suggestingthat at these stages tadpoles are capable of mounting an increasein stress hormone production in response to changes in the externalenvironment. Our results also highlight the importance of comparativestudies in understanding the development of the stress axis.     

Anatomical features of Leiopelma embryos and larvae: implications for anuran evolution

A controversial issue in anuran systematics is the relationship of Leiopelma to other anurans because recent phylogenetic constructions imply different relationships among the basal frogs. Of particular evolutionary interest is whether early development of Leiopelma resembles an ancestral salamander-like larva, an anuran tadpole, or neither. In the 1950s, Neville G. Stephenson hypothesized that direct development is the primary mode of development in amphibians, based on the fact that Leiopelma spp. lack a free-living (=feeding) larval stage. Although this hypothesis has not been generally accepted, it has not been formally refuted. We review Stephenson's work on Leiopelma and examine the anatomy of embryos/"larvae" of the four extant Leiopelma species for evidence of vestigial larval features that might refute the "direct-developing ancestor" hypothesis. We describe internal oral features in early developmental stages of Leiopelma and compare Leiopelma with a closely related basal anuran, Ascaphus, to assess whether their early developmental stages share any derived features. In Leiopelma hochstetteri, embryos/larvae have open gill slits and some faint rugosities around one gill slit that may be vestiges of gill rakers or filters. They also have more intestinal loops, indicative of an elongated alimentary tract, at earlier rather than late embryonic/larval stages. Collectively, these features support the view that the ancestor of Leiopelma had a free-swimming, free-feeding, aquatic larva. The palatoquadrate of Leiopelma archeyi reorients approximately 40 degrees from a more horizontal to a more vertical position through embryonic/"larval" development. This amount of cranial remodeling is intermediate between that seen in salamanders (17-27 degrees) and that reported for Ascaphus (64 degrees ) and other basal frogs (71-78 degrees) at metamorphosis. We found no internal oral features that Leiopelma shares specifically with Ascaphus. However, Leiopelma embryos have a ventrally positioned mouth and a downturned rostrum, characteristic of Ascaphus and other stream-adapted tadpoles.     

Multiple predator effects on size-dependent behavior and mortality of two species of anuran larvae

This study examined the effects of multiple predators on size‐specific behavior and mortality of two species of anuran larvae. Particularly, we focused on how trait changes in predators and prey may be transmitted to other species in the food web. In laboratory experiments, we examined the effects of bluegill sunfish, Lepomis macrochirus, and the odonate larva Anax junius on behavior and mortality of tadpoles of the bullfrog, Rana catesbeiana, and the green frog R. clamitans. Experiments were conducted with predators alone and together to assess effects on behavior and mortality of the tadpoles. The experiments were replicated on five size classes of the tadpoles to evaluate how responses varied with body size.
Predation rates by Anax were higher on bullfrogs than on green frogs, and both bullfrogs and green frogs suffered greater mortality from Anax than from bluegill. Bluegill only consumed green frogs. Predation rates by both predators decreased with increasing tadpole size and decreased in the non‐lethal (caged) presence of the other predator. Both anuran larvae decreased activity when exposed to predators. Bullfrogs, however, decreased activity more in the presence of Anax than in the presence of bluegill, whereas green frogs decreased activity similarly in the presence of both predators. The largest size class of green frogs, but not of bullfrogs, exhibited spatial avoidance of bluegill. These responses were directly related to the risk posed by the different predators to each anuran species. Anax activity (speed and move frequency) also was higher when alone than in the non‐lethal presence of bluegill. We observed decreased predation rate of each predator in the non‐lethal presence of the other, apparently caused by two different mechanisms. Bluegill decreased Anax mortality on tadpoles by restricting the Anax activity. In contrast, Anax decreased bluegill mortality on tadpoles by reducing tadpole activity. We discuss how the activity and spatial responses of the tadpoles interact with palatability and body size to create different mortality patterns in the prey species and the implications of these results to direct and indirect interactions in this system.     

Functional distance and establishment of non-native species with complex life cycles

More than 80% of animals have complex life cycles and undergo distinct changes in ecology and morphology during development. The strength and type of factors regulating each life-stage may differ as an organism may occupy different niches during ontogeny. We examined the functional distance at larval and adult life-stages of two non-native anurans (Green Tree Frog [Hyla cinerea] and Bullfrog [Lithobates catesbeianus]) that have established in a Chihuahuan Desert anuran assemblage in Big Bend National Park. Both life stages of both non-native species occupied niche space outside of the native assemblage. At the larval stage, the ability of the tadpoles to utilize permanent aquatic habitats and coexist with predatory fishes differentiated the non-native species from the majority of the native species that are restricted to temporary pools. At the post-metamorphic life stage, each species appears to have established by exploiting unoccupied habitat and trophic niches in the recipient community. The arboreal habits of H. cinerea may enable it to utilize resources in microhabitats that are otherwise not used by native species because arboreal frogs are absent from this native assemblage. The large body size of post-metamorphic L. catesbeianus may enable it to utilize larger food resources that are otherwise unavailable to the smaller-bodied natives. Separate comparison of larval and adult functional traits between non-natives and the native community may help predict their potential establishment or invasion success as well as aid in the development of stage-specific control or eradication efforts.     

A review of ecological interactions between native frogs and invasive cane toads in Australia

Translocated from their native range in the Americas in 1935, cane toads (Rhinella marina, Bufonidae) have now spread through much of tropical and subtropical Australia. The toad's invasion and impact have attracted detailed study. In this paper, I review information on ecological interactions between cane toads and Australian anurans. The phylogenetic relatedness and ecological similarity between frogs and toads creates opportunities for diverse interactions, ranging from predation to competition to parasite transfer, plus a host of indirect effects mediated via impacts of toads on other species, and by people's attempts to control toads. The most clear‐cut effect of toads on frogs is a positive one: reducing predator pressure by fatally poisoning anuran‐eating varanid lizards. However, toads also have a wide range of other effects on frogs, some positive (e.g. taking up parasites that would otherwise infect native frogs) and others negative (e.g. eating frogs, poisoning frogs, competing with tadpoles). Although information on such mechanisms predicts intense interactions between toads and frogs, field surveys show that cane toad invasion has negligible overall impacts on frog abundance. That counter‐intuitive result is because of a broad balancing of negative and positive impacts, coupled with stochastic (weather‐induced) fluctuations in anuran abundance that overwhelm any impacts of toads. Also, the impacts of toads on frogs differ among frog species and life‐history stages, and depend upon local environmental conditions. The impacts of native frogs on cane toads have attracted much less study, but may well be important: frogs may impose biotic resistance to cane toad colonization, especially via competition in the larval phase. Overall, the interactions between native frogs and invasive toads illustrate the diverse ways in which an invader's arrival can perturb the native fauna by both direct and indirect mechanisms, and by which the native species can curtail an invader's success. These studies also offer a cautionary tale about the difficulty of predicting the impact of an invasive species, even with a clear understanding of mechanisms of direct interaction.     

Common reed (Phragmites australis) invasion and amphibian distribution in freshwater wetlands

Invasive plants can substantially modify wetland structure and animal distribution patterns. In eastern North America, a Eurasian haplotype of the common reed (Phragmites australis, haplotype M) is invading wetlands. We studied the invasion of common reed in freshwater wetlands of an urbanized landscape and its effects on the distribution of amphibians at different life stages. Specifically, we hypothesized that the probability of reed invasion would be greatest in wetlands near anthropic disturbances. We predicted that the probability of desiccation at sampling stations increases with reed cover. Furthermore, we expected that wetlands invaded by common reed would have lower amphibian abundances, apparent survival, and rates of recruitment. We conducted trapping surveys to compare anuran assemblages of tadpoles, juveniles, and adults in 50 wetlands during two field seasons. The probability of reed invasion in wetlands increased with the cover of heavily-managed areas within 1,000 m and the distance to the nearest forest, but decreased with the length of roads within 1,000 m. The probability of station desiccation increased with reed cover. We found no evidence of a negative effect of reed presence on anuran population parameters, at any life stage. Landscape variables, such as the percent cover of forest or heavily-managed areas within a given radius from each wetland, influenced the abundance or the apparent survival of juvenile frogs and the abundance of ranid tadpoles. Our results show that amphibian patterns depend more strongly on the structure of the landscape surrounding wetlands than on exotic reed invasion in wetlands.     

The morphology of amphibian skin vascularization before and after metamorphosis

Summary The skin vascularization was investigated troughout the ontogenetic development and in adults of two anurans, Rana temporaria and Bufo bufo, and two urodeles, Triturus vulgaris and Triturus cristatus. It was found that, contrary to the urodele larvae, the anuran tadpoles have a very sparse skin vascularization. During the early stages of anuran metamorphosis the skin capillary network becomes dense; later, during skin metamorphosis, a second, venous, network is formed as anastomoses between the subcutaneous vein ramifications. In the urodeles, metamorphosis is not accompanied by any significant morphological changes in the skin vascularization, and a subcutaneous network is not formed. It is suggested that the reduced skin vascularization in anuran tadpoles is an advanced larval character relative to the rich vascularization of the skin in urodele larvae. It is further suggested that anuran tadpoles have a reduced ability to utilize gaseous exchange through the skin. The function of the subcutaneous venous network found in anurans after metamorphosis is obscure; experiments indicate a vasomotor regulation which is neither adrenergic nor cholinergic.Abbreviations a arteriole - A artery - an anastomosis between the subepidermal capillary network and the subcutaneous venous network - C stratum compactum - E epidermis - ec subepidermal capillaries - S stratum spongiosum - sv subcutaneous venous network - v venule - V vein - vc venae comitantes     

Ammonia as a pheromone in anuran tadpoles

In laboratory experiments, conspecific excretions and ammonia solutions evoked avoidance reactions in tadpoles of three anuran species, Bufo bufo, Rana temporaria, and Rana arvalis. A differential sensitivity of ammonia chemoreception was determined for two anuran species. For Bufo bufo tadpoles, these characteristics at ammonia background concentration of 0.2 mg/l lied in the range 150 % < dI/I < 500 %, and for background of 0.4 mg/l the value lied in the range 400 % < dI/I < 500 %. For Rana temporaria tadpoles, differential threshold against ammonia background concentration of 0.15 mg/l was close to 200 % and against background ammonia concentration of 1.1 mg/l was close to 100 %. These results suggest that such sensitivity of both anurans is sufficient for using ammonia in intra- and interspecies communication.     

Choice of optimal oviposition sites by Hoplobatrachus occipitalis (Anura: Ranidae) in an unpredictable and patchy environment

 The rock pools on the river bank of the Comoé National Park (West Africa) provide a very diverse and unpredictable environment for anuran larval development. Because rock pools differ considerably in biotic and abiotic parameters, it should be adaptive for reproducing anurans to choose the most suitable oviposition sites. During the beginning of each rainy season (March to May), from 1991 to 1995, we investigated the allocation of breeding sites by Hoplobatrachus occipitalis, counted the number of eggs, and measured several biotic and abiotic factors that might influence the choice of a spawning site. The probability of predation by conspecific cannibalistic tadpoles and the water-holding capacity (WHC) of pools were the best predictor of number of eggs laid. We experimentally investigated the influence of these two parameters on egg-laying and showed that adults potentially can assess the presence, density, and size of tadpoles in pools by chemical cues. Likewise, manipulation of the water-holding capacity caused a rapid change in egg-laying behavior. To assess the risk of desiccation, frogs have to visit familiar pools repeatedly to monitor the decrease in volume over time and thus gain information about the pools’ water-holding capacity. Received: 22 April 1996 / Accepted: 17 July 1996     

Lack of phylogenetic signal in the variation in anuran microhabitat use in southeastern Brazil

Microhabitat use is an important component of anuran behavior in both the tadpole and the adult stages. It is potentially influenced by phylogeny and extant ecological factors acting as selective pressures, such as predation, competition, or physical habitat properties. We aimed to test whether patterns of microhabitat use vary among species, habitats and sites, and how much of this variation can be explained by phylogenetic relatedness. We collected data on microhabitat use at five different sites, where we obtained a total of 4,230 records of individual tadpoles of 34 species in 15 genera and 7 families, and a total of 1,163 records of adult individuals of 39 species in 16 genera and 8 families. Mantel tests conducted to relate species dissimilarities in microhabitat use and phylogenetic relatedness indicated a weak but significant relationship for adult anurans, and no relationship for tadpoles. Our results suggest that microhabitat use is a plastic and variable trait, overcoming phylogenetic signal in tadpoles. In adult anurans, very little of the variation in microhabitat use can be explained by phylogenetic relatedness. Microhabitat use is not a good predictor of phylogeny, but it may be a very interesting subject to study natural selection and adaptation.     

The role of trematode parasites in larval anuran communities: an aquatic ecologist’s guide to the major players

Conservation strategies depend on our understanding of the ecosystem and community dynamics. To date, such understanding has focused mostly on predator–prey and competitor interactions. It is increasingly clear, however, that parasite–host interactions may represent a large, and important, component of natural communities. The need to consider multiple factors and their synergistic interactions if we are to elucidate the contribution of anthropogenic factors to loss in biodiversity is exemplified by research into present-day amphibian declines. Only recently has the role of factors such as trematode parasite infections been incorporated into studies of the population and community dynamics of aquatic systems. We argue that this is due, at least in part, to difficulties faced by aquatic ecologists in sifting through the complex systematics that pervade the parasite literature. We note that two trematode species are of dominant importance with regard to North American larval anuran communities, and provide in this review a clear explanation of how to distinguish between the infective stages of these two parasites. We describe the general biology and life history of these parasites, as well as what is known about their effect on larval anurans, and the interactive effects of environmental stressors (typically anthropogenic in nature) and parasites on larval anurans. We hope that this review will convince the reader of the potential importance of these parasites to aquatic communities in general, and to amphibian communities specifically, and will also provide the information necessary for aquatic ecologists to more frequently consider the role of these parasites in their studies of aquatic ecology.     

Extra-intestinal localization of Goussia sp. (Apicomplexa) oocysts in Rana dalmatina (Anura: Ranidae), and the fate of infection after metamorphosis

Although coccidia of the genus Goussia are common parasites of fish, only 2 species have been described in amphibians: G. hyperolisi from common reed frogs Hyperolius viridiflavus from Kenya and G. neglecta from unspecified European water frogs of the genus Rana from Germany. The genus Goussia is characterized by an oocyst, with a fine oocyst wall, containing 4 dizoic sporocysts that are composed of 2 valves joined by a longitudinal suture and lacking a Stieda body (typical for the genus Eimeria). To date, infections in amphibians were generally considered to be specific to the intestine of aquatic larval stages (tadpoles) of anurans. Herein, we report on: (1) the presence of oocysts of Goussia sp. in an extra-intestinal location (liver) of tadpoles of the agile frog R. dalmatina and (2) the presence of oocysts in the liver of both juvenile and subadult R. dalmatina. These observations represent novel traits for Goussia infections in amphibians; they may explain the vertical transmission of Goussia in tadpoles.     

Do prostaglandins regulate external gill regression in anurans?

Although the endocrinological mechanism controlling regression of the internal, larval gills of anurans (frogs and toads) is well understood, the mechanism regulating loss of the external, embryonic gills is not known. Based on the homology of the mammalian ductus arteriosus with a portion of the amphibian branchial arches, and the regulation of blood flow in the mammalian ductus by prostaglandins of the E family (PGEs), we hypothesized that anuran external gill loss is also regulated by PGEs. To test this hypothesis, we topically applied both PGE2 and a synthetic analogue of PGE1, misoprostol, to embryos and young hatchlings of the red-eyed treefrog, Agalychnis callidryas. Both agents accelerated external gill regression. Furthermore, misoprostol overrode the inhibitory effect of hypoxia on gill regression in hatchlings and induced rapid loss of external gills in embryos, which normally maintain the gills until hatching. These observations support the hypothesis that PGEs regulate anuran external gill loss. The specific site of action for prostaglandins within the gills is not known; however, PGEs are secreted in the oral mucus of tadpoles, and this could be a natural topical source for these agents. PGEs offer a tool for manipulation of external gills and should facilitate tests of the physiological importance of these structures.     

Evolutionary causes and consequences of sequential polyandry in anuran amphibians

Among anuran amphibians (frogs and toads), there are two types of polyandry: simultaneous polyandry, where sperm from multiple males compete to fertilize eggs, and sequential polyandry, where eggs from a single female are fertilized by multiple males in a series of temporally separate mating events, and sperm competition is absent. Here we review the occurrence of sequential polyandry in anuran amphibians, outline theoretical explanations for the evolution of this mating system and discuss potential evolutionary implications. Sequential polyandry has been reported in a limited number of anurans, but its widespread taxonomic and geographic distribution suggests it may be common. There have been no empirical studies that have explicitly investigated the evolutionary consequences of sequential polyandry in anurans, but species with this mating pattern share an array of behavioural, morphological and physiological characteristics, suggesting that there has been common sexual selection on their reproductive system. Sequential polyandry may have a number of adaptive benefits, including spreading the risk of brood failure in unpredictable environments, insuring against male infertility, or providing genetic benefits, either through good genes, intrinsic compatibility or genetic diversity effects. Anurans with sequential polyandry provide untapped opportunities for innovative research approaches that will contribute significantly to understanding anuran evolution and also, more broadly, to the development of sexual‐selection and life‐history theory.     

Trait Performance Correlations across Life Stages under Environmental Stress Conditions in the Common Frog,Rana temporaria

If an organism''s juvenile and adult life stages inhabit different environments, certain traits may need to be independently adapted to each environment. In many organisms, a move to a different environment during ontogeny is accompanied by metamorphosis. In such organisms phenotypic induction early in ontogeny can affect later phenotypes. In laboratory experiments we first investigated correlations between body morphology and the locomotor performance traits expressed in different life stages of the common frog, Rana temporaria: swimming speed and acceleration in tadpoles; and jump-distance in froglets. We then tested for correlations between these performances across life stages. We also subjected tadpoles to unchanging or decreasing water levels to explore whether decreasing water levels might induce any carry-over effects. Body morphology and performance were correlated in tadpoles; morphology and performance were correlated in froglets: hence body shape and morphology affect performance within each life stage. However, performance was decoupled across life stages, as there was no correlation between performance in tadpoles and performance in froglets. While size did not influence tadpole performance, it was correlated with performance of the metamorphosed froglets. Experiencing decreasing water levels accelerated development time, which resulted in smaller tadpoles and froglets, i.e., a carry-over effect. Interestingly, decreasing water levels positively affected the performance of tadpoles, but negatively affected froglet performance. Our results suggest that performance does not necessarily have to be correlated between life stages. However, froglet performance is size dependent and carried over from the tadpole stage, suggesting that some important size-dependent characters cannot be decoupled via metamorphosis.     

How trematodes cause limb deformities in amphibians

We used trematode cyst infestation to induce limb deformities in two species of frogs of the genus Rana and compared them to deformities induced by surgical limb bud rotations. The specific deformities produced by both treatments closely resemble those of wild-caught deformed amphibians and are consistent with a known developmental response to disruption of the spatial organization of cells in developing limb buds. Histological analysis showed that trematode cysts cause massive disruption and abnormal cellular growth involving the limb buds of infected individuals. Our results indicate that trematode cyst infestation causes deformities in frogs by perturbation of the positional relationships of cells in developing limb buds. The crippling effects of cyst-infection on frogs may reflect complex co-evolutionary interactions among trematodes, frogs, and other hosts in the trematode's life cycle.     

The evolution of reproductive diversity in Afrobatrachia: A phylogenetic comparative analysis of an extensive radiation of African frogs

The reproductive modes of anurans (frogs and toads) are the most diverse of terrestrial vertebrates, and a major challenge is identifying selective factors that promote the evolution or retention of reproductive modes across clades. Terrestrialized anuran breeding strategies have evolved repeatedly from the plesiomorphic fully aquatic reproductive mode, a process thought to occur through intermediate reproductive stages. Several selective forces have been proposed for the evolution of terrestrialized reproductive traits, but factors such as water systems and co‐evolution with ecomorphologies have not been investigated. We examined these topics in a comparative phylogenetic framework using Afrobatrachian frogs, an ecologically and reproductively diverse clade representing more than half of the total frog diversity found in Africa (～400 species). We infer direct development has evolved twice independently from terrestrialized reproductive modes involving subterranean or terrestrial oviposition, supporting evolution through intermediate stages. We also detect associations between specific ecomorphologies and oviposition sites, and demonstrate arboreal species exhibit an overall shift toward using lentic water systems for breeding. These results indicate that changes in microhabitat use associated with ecomorphology, which allow access to novel sites for reproductive behavior, oviposition, or larval development, may also promote reproductive mode diversity in anurans.