Osseous pathologies in the lungless salamander Desmognathus fuscus (Plethodontidae)

The majority of reported pathologies in lissamphibians (salamanders, caecilians and frogs) include limb deformities such as missing limbs, multiple extra limbs and digits, or incomplete limb formation. However, comparatively little is known about congenital vertebral malformations or posttraumatic pathologies (e.g. injuries, infections) in the vertebral column of salamanders. In the present study, we describe eight vertebral deformities in three cleared and stained specimens of Desmognathus fuscus. Two specimens display developmental deformities which range from a potential non-segmented wedge vertebra to fully segmented hemivertebrae. The vertebral pathology in the third specimens possibly results from a parasitic infection. Apparently, these osseous deformities were not severe enough to prohibit survival of the specimens.     

Explanation for naturally occurring supernumerary limbs in amphibians

The occasional occurrence of high frequencies of limb abnormalities, including extra limbs, in natural populations of amphibians has long been a puzzle. In this paper we report the discovery of a population in which such limb abnormalities appear to be caused by a parasitic flatworm (trematode) that uses amphibians as intermediate hosts. The cercarial larval stage of the trematode attacks amphibians, penetrating the skin to form cysts (metacercariae). The cysts are preferentially localized in the cloacal region, including the developing hind limb regions in larvae of both frogs (Hyla regilla) and salamanders (Ambystoma macrodactylum). A wide range of limb abnormalities are seen, including duplicated limb structures ranging from extra digits to several extra whole limbs. We hypothesize that these limb abnormalities result from localized regulatory responses of developing and regenerating limb tissues to mechanical disruption caused by the trematode cysts. We have tested this idea by implanting inert resin beads into developing limb buds of frogs and salamanders. Since this treatment can cause supernumerary limb structures, our hypothesis is sufficient to explain the naturally occurring extra limbs.     

Assessment of rates of deformity in wild frog populations using in situ cages: a case study of Leopard Frogs (Rana pipiens) in Ontario, Canada

High rates of deformity in wild amphibian populations from north-eastern North America have been increasingly reported since 1995. In the St Lawrence River basin (Canada) elevated frequencies of limb and eye deformities in mudpuppies (Necturus maculosus) and leopard frogs (Rana pipiens) were recorded in the early 1990s. A caging study was conducted during 1998 to verify the rates recorded in leopard frogs and pursue the potential causes of deformities seen in juveniles and adults. Week-old leopard frog tadpoles were collected from a reference wetland and maintained through to metamorphosis in cages in previously identified high risk wetlands. Deformity frequencies were measured and compared with frequencies measured in wild populations of leopard frogs inhabiting the same wetlands. The results of caging studies and sampling of wild populations were also compared with corresponding data collected from a reference wetland. No deformities were observed in caged or wild reference animals. Very low deformity frequencies (up to 2.2%) were observed in frogs caged in high risk wetlands, but greater frequencies (3.4-10%) were observed in wild young-of-the-year frogs captured at the same sites. The types of deformities were similar among groups; they included fused, missing or extra digits and disproportionate hindlimb length or eye pupil size. In addition, mortality rates were elevated in two cages in high risk wetlands. In general, the caging procedure was effective in establishing the potential for production of deformities in the waters of a given wetland, but tended to underestimate the rates calculated for samples of wild populations. The ramifications of the first-year findings for similar assessments of amphibian deformity rates and establishment of cause-effect linkages are discussed.     

Hind limb malformations in free-living northern leopard frogs (Rana pipiens) from Maine, Minnesota, and Vermont suggest multiple etiologies

BACKGROUND: Reports of malformed frogs have increased throughout the North American continent in recent years. Most of the observed malformations have involved the hind limbs. The goal of this study was to accurately characterize the hind limb malformations in wild frogs as an important step toward understanding the possible etiologies. METHODS: During 1997 and 1998, 182 recently metamorphosed northern leopard frogs (Rana pipiens) were collected from Minnesota, Vermont, and Maine. Malformed hind limbs were present in 157 (86%) of these frogs, which underwent necropsy and radiographic evaluation at the National Wildlife Health Center. These malformations are described in detail and classified into four major categories: (1) no limb (amelia); (2) multiple limbs or limb elements (polymelia, polydactyly, polyphalangy); (3) reduced limb segments or elements (phocomelia, ectromelia, ectrodactyly, and brachydactyly; and (4) distally complete but malformed limb (bone rotations, bridging, skin webbing, and micromelia). RESULTS: Amelia and reduced segments and/or elements were the most common finding. Frogs with bilateral hind limb malformations were not common, and in only eight of these 22 frogs were the malformations symmetrical. Malformations of a given type tended to occur in frogs collected from the same site, but the types of malformations varied widely among all three states, and between study sites within Minnesota. CONCLUSIONS: Clustering of malformation type suggests that developmental events may produce a variety of phenotypes depending on the timing, sequence, and severity of the environmental insult. Hind limb malformations in free-living frogs transcend current mechanistic explanations of tetrapod limb development.     

The role of habitat in structuring Halipegus occidualis metapopulations in the green frog

The transmission dynamics of the trematode Halipegus occidualis in its definitive host, Rana clamitans, have been examined over a 5-yr period in a North Carolina pond. The breeding season of green frogs coincides with the period of worm recruitment, during which time male frogs are territorial and females show strong site fidelity. This site fidelity allows inferences to be made regarding the suitability of a particular habitat for worm transmission based on frog infection intensities within that habitat. Four foci of infection were identified in the pond by plotting worm infrapopulation size against site of host capture. Sites within infection foci are characterized by shallow water and emergent vegetation, factors favorable for overlapping distributions of the 4 hosts in the life cycle of H. occidualis. Consistent year-to-year worm prevalences and intensities, despite fluctuations in frog population size, are thought to be the result of a relatively constant proportion of the frog population being present in infection foci each year. Removal of worms from heavily infected frogs in the fifth year resulted in further heavy worm recruitment by treated frogs suggesting that site selection can predispose a frog to heavy infection. Further, the sum of removed parasites and those recruited after parasite removal by treated frog hosts was higher than worm infrapopulations observed in previous years, indicating that worm density regulates parasite infrapopulation size in heavily infected frogs.     

Vitamin A induced homeotic hindlimb formation on dorsal and ventral sides of regenerating tissue of amputated tails of Japanese brown frog tadpoles

When anuran tadpoles are treated with vitamin A after tail amputation, hindlimb‐like structures can be generated instead of the lost tail part at the amputation site. This homeotic transformation was initially expected to be a key to understanding the body plan of vertebrates. Unfortunately, homeotic limb formation has been reproduced in only some Indian frog species and a European species, but not in experimental anurans such as Xenopus laevis or Rana catesbeiana. Consequently, this fascinating phenomenon has not been well analyzed, especially at the molecular level. In addition, the initial processes of ectopic limb development are also unclear because morphological changes in the early phases have not been analyzed in detail. In this study, we report the induction of homeotic transformation using Japanese brown frogs and present a detailed morphological analysis. Unexpectedly, the ectopic limbs developed not only at the ventral sites, but also at the dorsal sites of the tail regenerates of vitamin A‐treated tadpoles. The relationship between position and axial orientation of ectopic limbs suggested the double duplication of positional value order along the rostral‐caudal axis and the dorsal‐ventral axis of the tail regenerates.     

Whole animal measurements of shear and adhesive forces in adult tree frogs: insights into underlying mechanisms of adhesion obtained from studying the effects of size and scale

This allometric study of adhesion in 15 Trinidadian tree frog species investigates how relationships between length, area and mass limit the ability of adult frog species of different sizes to adhere to inclined and overhanging surfaces. Our experiments show that hylid frogs possess an area-based wet adhesive system in which larger species are lighter than expected from isometry and adhere better than expected from their toe pad area. However, in spite of these adaptations, larger species adhere less well than smaller species. In addition to these adhesive forces, tree frogs also generate significant shear forces that scale with mass, suggesting that they are frictional forces. Toe pads detach by peeling and frogs have strategies to prevent peeling from taking place while they are adhering to surfaces, including orienting themselves head-up on slopes. The scaling of tree frog adhesion is also used to distinguish between different models for adhesion, including classic formulae for capillarity and Stefan adhesion. These classic equations grossly overestimate the adhesive forces that tree frogs produce. More promising are peeling models, designed to predict the pull-off forces of adhesive tape. However, more work is required before we can qualitatively and quantitatively describe the adhesive mechanism of tree frogs.     

The evolution of development: two portraits of skull ossification in pipoid frogs

Abstract.— Development creates morphology, and the study of developmental processes has repeatedly shed light on patterns of morphological evolution. However, development itself evolves as well, often concomitantly with changes in life history or in morphology. In this paper, two approaches are used to examine the evolution of skull development in pipoid frogs. Pipoids have highly unusual morphologies and life histories compared to other frogs, and their development also proves to be remarkable. First, a phylogenetic examination of skull bone ossification sequences reveals that jaw ossification occurs significantly earlier in pipoids than in other frogs; this represents a reversal to the primitive vertebrate condition. Early jaw ossification in pipoids is hypothesized to result from the absence of certain larval specializations possessed by other frogs, combined with unusual larval feeding behaviors. Second, thin-plate spline morphometric studies of ontogenetic shape change reveal important differences between pipoid skull development and that of other frogs. In the course of frog evolution, there has been a shift away from salamander-like patterns of ontogenetic shape change. The pipoids represent the culmination of this trend, and their morphologies are highly derived in numerous respects. This study represents the first detailed examination of the evolution of skull development in a diverse vertebrate clade within a phylogenetic framework. It is also the first study to examine ossification sequences across vertebrates, and the first to use thin-plate spline morphometrics to quantitatively describe ontogenetic trajectories.     

Towards a Better Understanding of the Use of Probiotics for Preventing Chytridiomycosis in Panamanian Golden Frogs

Populations of native Panamanian golden frogs (Atelopus zeteki) have collapsed due to a recent chytridiomycosis epidemic. Reintroduction efforts from captive assurance colonies are unlikely to be successful without the development of methods to control chytridiomycosis in the wild. In an effort to develop a protective treatment regimen, we treated golden frogs with Janthinobacterium lividum, a skin bacterium that has been used to experimentally prevent chytridiomycosis in North American amphibians. Although J. lividum appeared to colonize A. zeteki skin temporarily, it did not prevent or delay mortality in A. zeteki exposed to Batrachochytrium dendrobatidis, the causative agent of chytridiomycosis. After introduction of J. lividum, average bacterial cell counts reached a peak of 1.7 × 10(6) cells per frog ~2 weeks after treatment but declined steadily after that. When J. lividum numbers declined to ~2.8 × 10(5) cells per frog, B. dendrobatidis infection intensity increased to greater than 13,000 zoospore equivalents per frog. At this point, frogs began to die of chytridiomycosis. Future research will concentrate on isolating and testing antifungal bacterial species from Panama that may be more compatible with Atelopus skin.     

Determination of age, longevity and age at reproduction of the frogMicrohyla ornata by skeletochronology

Skeletochronological estimation of age, longevity, age at sexual maturity and breeding of Microhyla ornata was done. Frogs (n=62) were collected locally in August (rainy season) 1997 and brought to the laboratory. Body mass and snout-vent-length (SVL) of each frog was recorded; the 4th toe of both the hind limbs was clipped under anaesthesia, fixed in 10% formalin, demineralized in 5% nitric acid and processed for histology. Limb bones (femur, humerus, tibiofibula and radioulna) of 6 large sized frogs were also processed for skeletochronology in order to study the rate of resorption. Gonads of 25 frogs (belonging to different body size ranges) were processed for histology in order to ascertain the gametogenic status of individual frogs. One to four growth rings consisting of growth zones and lines of arrested growth (LAGs) were noticed in frogs of different body sizes; the number of LAGs remained identical in all the limb bones and phalanges in 5 out of 6 frogs. Back calculation indicated that the resorption rate is very low in this frog. Male frogs possessed sperm bundles in seminiferous tubules in the 1st year, while females showed yolky follicles in the ovary in the 2nd year. Frogs found in amplexus were 3 5 years old. The results suggest that this frog may live for a maximum of 5 years in the natural population.     

Studies on the appearance of skeletal anomalies in red porgy: effect of culture intensiveness,feeding habits and nutritional quality of live preys

Despite the great interest of red porgy as a new species for Mediterranean aquaculture, its commercial production is constrained by the high incidence of skeletal deformities occurring in this species under culture conditions. Several studies have been conducted to better understand the origin of these anomalies in this species, using different system intensiveness, rotifers enrichment products or rotifers docosahexaenoic acid content. The first study showed that culture intensification increased the number of fish with an extra vertebrae, what was probably related to the different nutritional quality of live preys employed in each treatment, since water temperature, salinity and genetic background were identical for the different batches of fish studied. Total incidence of skeletal abnormalities was higher in the intensive system, particularly cranial abnormalities and kyphosis in the cephalic vertebrae. In both rearing systems the most common skeletal anomalies were vertebral column disorders, lordosis and fused vertebrae, their localization along the column being affected by the culture intensiveness. Rotifer enrichment, predominantly its docosahexaenoic acid content significantly affected deformities occurrence. A marked positive effect of rotifer docosahexaenoic acid content was found on larval survival. X‐ray studies denoted elevated levels of bone abnormalities associated, in both trials, to low docosahexaenoic acid content in live preys. Among different anomalies, the presence of fused vertebrae was the most frequent deformity for both rearing trials. A 50% reduction in the number of deformed fish for each type of deformity was obtained when the larvae were fed higher docosahexaenoic acid levels, denoting the important role of this fatty acid in bone development. Further studies are needed to elucidate the importance of essential fatty acids on the development of bone deformities in fish, since the functions of these fatty acids differ among them and can lead to very different effects in fish metabolism, including bone formation.     

吉林产哈士蟆中氨基酸含量的研究

应用日立835—50型氨基酸自动分析仪,对哈士蟆十种干样品进行了氨基酸成分的比较分析,结果表明:①必需氨基酸含量由高至低依次为雄蛙腿肉,雌蛙腿肉、卵、整只雄蛙、整只雌蛙、雄蛙皮、哈士蟆油、雌蛙皮、雌蛙腿骨、雄蛙腿骨。②氨基酸总量由高至低依次为雄蛙腿肉、雌蛙腿肉、雄蛙皮、雌蛙皮、卵、整只雄蛙、整只雌蛙、哈士蟆油、雌蛙腿骨、雄蛙腿骨。③雄蛙腿肉和雌蛙腿肉中各种氨基酸含量由大到小的排列顺序基本一致。     

Past and Present Risk: Exposure to Predator Chemical Cues Before and after Metamorphosis Influences Juvenile Wood Frog Behavior

Animals are exposed to different predators over their lifespan. This raises the question of whether exposure to predation risk in an early life stage affects the response to predators in subsequent life stages. In this study, we used wood frogs (Rana sylvatica) to test whether exposure to cues indicating predation risk from dragonfly larvae during the wood frog larval stage affected post‐metamorphic activity level and avoidance of garter snake chemical cues. Dragonfly larvae prey upon wood frogs only during the larval stage, whereas garter snakes prey upon wood frogs during both the larval stage and the post‐metamorphic stage. Exposure to predation risk from dragonflies during the larval stage caused post‐metamorphic wood frog juveniles to have greater terrestrial activity than juvenile wood frogs that were not exposed to larval‐stage predation risk from dragonflies. However, exposure to predation risk as larvae did not affect juvenile wood frog responses to chemical cues from garter snakes. Wood frogs exposed as larvae to predation risk from dragonfly larvae avoided garter snake chemical cues to the same extent as wood frog larvae not exposed to predation risk from dragonfly larvae. Our results demonstrate that while some general behaviors exhibit carry‐over effects from earlier life stages, behavioral responses to predators may remain independent of conditions experienced in earlier life stages.     

Protein synthesis in the organs and tissues of frogs with local exposure to low temperatures]

Intensity of synthesis of total proteins and spectrum of synthesized proteins has been studied in various organs and tissues of sharp-faced frogs during local profound cooling of one of its limbs. It was shown that the effect of low temperatures on a certain part of a body of a poikilothermic animal has induced both a response of the whole organism and a much more acute response of the cooled tissues. The organism responded by a fall of the whole body temperature and a decrease in the activity of synthesis of total proteins of the viscera. Local cooling of a part of the frog body has induced a more significant decrease in the protein synthesis intensity in the cooled tissues alteration in the spectrum of synthesized proteins and appearance of specific proteins.     

How the environment shapes animal signals: a test of the acoustic adaptation hypothesis in frogs

Long‐distance acoustic signals are widely used in animal communication systems and, in many cases, are essential for reproduction. The acoustic adaptation hypothesis (AAH) implies that acoustic signals should be selected for further transmission and better content integrity under the acoustic constraints of the habitat in which they are produced. In this study, we test predictions derived from the AAH in frogs. Specifically, we focus on the difference between torrent frogs and frogs calling in less noisy habitats. Torrents produce sounds that can mask frog vocalizations and constitute a major acoustic constraint on call evolution. We combine data collected in the field, material from scientific collections and the literature for a total of 79 primarily Asian species, of the families Ranidae, Rhacophoridae, Dicroglossidae and Microhylidae. Using phylogenetic comparative methods and including morphological and environmental potential confounding factors, we investigate putatively adaptive call features in torrent frogs. We use broad habitat categories as well as fine‐scale habitat measurements and test their correlation with six call characteristics. We find mixed support for the AAH. Spectral features of torrent frog calls are different from those of frogs calling in other habitats and are related to ambient noise levels, as predicted by the AAH. However, temporal call features do not seem to be shaped by the frogs’ calling habitats. Our results underline both the complexity of call evolution and the need to consider multiple factors when investigating this issue.     

Role of thyroid hormone and retinoid receptors in the homeotic transformation of tails into limbs in frogs

We provide here further data on the dramatic homeotic transformation of tails into limbs which is induced by retinoids during frog tadpole tail regeneration. The effect can still be produced up to nine days after tail amputation by which time tail regeneration has essentially been completed. Complete tail amputation is needed for the effects to be manifest, partial damage of various sorts to the tail is not enough. We show that as well as retinyl palmitate, other retinoids such as all-trans-retinoic acid and TTNPB, which is a RAR specific retinoid, can induce the homeotic transformation. TTNPB has a 300x greater potency than retinoic acid. Prolactin, which inhibits thyroid hormone production, prevents the appearance of limbs on the tail from which we conclude that thyroid hormone is needed. We present preliminary evidence from RT-PCR that all six retinoid receptors, the three retinoic acid receptors (RARs), and the three retinoid X receptors (RXRs), are present in the normal tail blastema and that after retinoid treatment RARα, RXRα, and RXRβ may be up-regulated. Finally, we show that when RA synthesis is inhibited, normal tail regeneration is inhibited. We conclude that tail regeneration depends upon a particular endogenous level of RA, but that when this level is raised by external administration and thyroid hormone receptors are present the up-regulation of certain retinoid receptors allows novel nuclear receptor interactions which results in the induction of limb-specific genes leading to the appearance of limbs on the tail. © 1996 Wiley-Liss, Inc.     

Population Patterns of a Riparian Frog (Rana swinhoana) Before and After an Earthquake in Subtropical Taiwan

We compared the population dynamics of a riparian ranid frog, Rana swinhoana, before (1996–1999) and after (1999–2001) a strong earthquake. This earthquake caused little disturbance to the vegetation and landscape of the study site but the stream and ponds dried up within a week. Nearly all frogs marked (1002 of 1004) before the earthquake had disappeared after the earthquake. Smaller, unmarked frogs began to appear in stream habitats about 9 mo after the earthquake, and the frog population was much smaller than it was before the earthquake. Population dynamics and temporal and spatial distribution of frogs before and after the earthquake correlated closely with the hydrology of the stream and ponds. The movement patterns of frogs before and after the earthquake were similar, suggesting frog behavior did not change in response to drastic changes in hydrology, and frogs continued to exhibit strong site-fidelity. Following the earthquake, stream water volume was much lower, especially in the summer, which allowed the normally winter-breeding frogs to breed year-round. Results demonstrate that a population of R. swinhoana can disappear suddenly as the result of a natural disturbance. We propose that anuran species that exhibit strong site-fidelity are particularly susceptible to extirpation of local populations because frogs may lack the behavioral plasticity to respond to sudden water depletion.     

Spawn site selection and colony size of the frog (Rana temporaria) and the toad (Bufo bufo)

Information has been collected from various sources on spawn site selection and colony size of the frog ( Rana temporaria ) and the toad ( Bufo bufo ). Details are tabulated for 764 frog breeding sites in Britain and in the Republic of Ireland and for 139 toad sites in Britain (the toad does not occur in Ireland). Both species tend to breed in whatever types of site are most available. In Britain the garden pond has steadily increased in importance for both species since 1950. Frogs usually spawned in shallow water, 266 reports out of a total of 588 (45%) referring to water up to six inches (15 cm) deep. Toads tended to spawn in deeper water, 48 reports out of 82 (59%) mentioning water 7–18 inches (18–46 cm) in depth. Median depths were: frog, seven inches (18 cm); toad 13 inches (33 cm). Information on colony size was assimilated for 574 frog colonies and 86 toad colonies. Colonies in gardens were smaller than those in other types of site. Relatively fewer large frog colonies (≡100 animals) were found to occur in London than elsewhere in Britain, and British colonies were generally larger than those in Ireland. Colonies of 100 or more individuals were recorded relatively more frequently for toads than for frogs. By combining information on choice of site and colony size, rough estimates can be derived of the percentage of frogs and toads breeding in gardens and other types of site. For instance, in the London area in the 1960s perhaps 20–25% of the frogs bred in gardens and private grounds. The ecological significance of these observations is discussed.