Histo-morphology of the thyroid gland during the larval development of Pleurodema borellii (Anura,Leptodactylidae)

The thyroid gland is essential in anurans, since thyroid hormones (TH) are the main regulators of larval development. Its absence or inactivity interrupts development and precludes metamorphosis. Histological changes are important diagnostic criteria for evaluating thyroid gland activity. However, there is a large larval diversity where the development of the thyroid gland development has not been studied. Pleurodema borellii is an anuran from northwest of Argentina with typical omnivorous pond tadpoles that can be easily raised in captivity. This study explores the histo-morphological changes of the thyroid gland architecture during larval development. Histological parameters indicate peak glandular activity in parallel with the intensity of the metamorphic transformations. These parameters regress towards the end of metamorphosis, indicating low TH release. P. borellii's thyroid gland does not appear to have relevant activity in post-metamorphic juvenile stages. This study is a first step towards understanding endocrine regulation during the development of Pleurodema borellii, and a reference to future studies in this species involving thyroid-dependent processes.     

Intraspecific developmental variation in the life cycle of the Andean Treefrog (Boana riojana): A temporal analysis

Intraspecific variation during the anuran larval period has been analyzed mainly in relation to the timing of metamorphosis and body size at metamorphosis. However, other traits may vary as well. We examined two developmental series of Boana riojana from the same population in two consecutive years and describe intraspecific variation in larvae of this species. We discuss how variation, if present, may influence its life cycle. We found that both larval series differed in the larval period length, one twice as long as the other. This variation primarily depended on when breeding occurred, metamorphosis was achieved during late spring in both generations and at similar sizes, and only the rate of larval development during premetamorphosis varied extensively between years. This is consistent with thyroid gland activity because when it became active the developmental trajectory became more canalized. No variation of staging sequence occurred in relation to the different durations of the larval period. However, in the long-lasting series we found two different morphs. Also, integument, thyroid gland, skeleton, and testis differentiation events occurred at the same developing stages. In contrast, ovarian differentiation proceeded at the same absolute age in both series. Sexual dimorphism becomes evident within the year after metamorphosis. The intraspecific heterochrony that we describe for the larval development of B. riojana does not lead to phenotypic variation at the end of metamorphosis. We discuss the importance of analyzing growth and development independently. Each proceeds differently in time, but with an interdependence at some point, because size and shape do not vary at the end of metamorphosis.     

Ontogeny of the Thyroid Glands During Larval Development of South American Horned Frogs (Anura,Ceratophryidae)

The role of thyroid hormone (TH) in anuran metamorphosis has been documented from a variety of approaches, but the sequence of morpho-histological development of the thyroid glands that produce the secretion of the hormone was assumed invariant from studies of relatively few species even when the effects of environmental influences on larval development and metamorphosis have been largely documented. There are anurans in which developmental and growth rates diverge, and the resulting heterochrony in growth and development produces giant/miniature tadpoles, and or rapid/delayed metamorphosis suggesting changes of the activity of the thyroid glands during larval development. Herein, we analyze the morpho-histological variation of the thyroid glands in larval series of Ceratophrys cranwelli, Chacophrys pierottii, Lepidobatrachus laevis and L. llanensis that share breeding sites along semiarid environments of the Chaco in South America, belong to a monophyletic lineage, and present accelerated patterns in growth and development in order to have a morphological evidence about a possible shift of TH physiology. We describe gross morphology and histology of the thyroid glands and find features shared by all studied species such as the presence of supernumerary heterotopic follicles; changes in the volume and number of follicles towards the metamorphic climax, and cuboidal epithelia with occasional intra-cellular vacuoles as signs of low glandular activity without a manifest peak at the climax as it was assumed for anurans. We discuss different lines of evidence to interpret sources of extra supplement of TH to support the rapid metamorphosis. These interpretations highlight the necessity to design a research program to investigate the endocrine variation during development of ceratophryids taking in account their morphology, physiology and ecology in order to learn more about the effects of environmental and developmental interactions involved in the anuran evolution.     

Heterochrony in Growth and Development in Anurans from the Chaco of South America

Heterochrony refers to those permutations in timing of differentiation events, and those changes in rates of growth and development through which morphological changes and novelties originate during phyletic evolution. This research analyzes morphological variation during the ontogeny of 18 different anuran species that inhabit semi-arid environments of the Chaco in South America. I use field data, collection samples, and anatomical methods to compare larval growth, and sequences of ontogenetic events. Most species present a similar pattern of larval development, with a size at metamorphosis related to the duration of larval period, and disappearance and transformations of larval features that occur in a short period between forelimb emergence and tail loss. Among these 18 species, Pseudis paradoxa has giant tadpole and long larval development that are the results of deviations of rates of growth. In this species events of differentiation that usually occur at postmetamorphic stages have an offset when tail is still present. Tadpoles of Lepidobatrachus spp. reach large sizes at metamorphosis by accelerate developmental rates and exhibit an early onset of metamorphic features. The uniqueness of the ontogeny of Lepidobatrachus indicates that evolution of anuran larval development may occasionally involve mid-metamorphic morphologies conserving a free feeding tadpole and reduction of the morphological-ecological differences between tadpoles and adults.     

Mechanisms and Consequences of Developmental Acceleration in Tadpoles Responding to Pond Drying

Many amphibian species exploit temporary or even ephemeral aquatic habitats for reproduction by maximising larval growth under benign conditions but accelerating development to rapidly undergo metamorphosis when at risk of desiccation from pond drying. Here we determine mechanisms enabling developmental acceleration in response to decreased water levels in western spadefoot toad tadpoles (Pelobates cultripes), a species with long larval periods and large size at metamorphosis but with a high degree of developmental plasticity. We found that P. cultripes tadpoles can shorten their larval period by an average of 30% in response to reduced water levels. We show that such developmental acceleration was achieved via increased endogenous levels of corticosterone and thyroid hormone, which act synergistically to achieve metamorphosis, and also by increased expression of the thyroid hormone receptor TRΒ, which increases tissue sensitivity and responsivity to thyroid hormone. However, developmental acceleration had morphological and physiological consequences. In addition to resulting in smaller juveniles with proportionately shorter limbs, tadpoles exposed to decreased water levels incurred oxidative stress, indicated by increased activity of the antioxidant enzymes catalase, superoxide dismutase, and gluthatione peroxidase. Such increases were apparently sufficient to neutralise the oxidative damage caused by presumed increased metabolic activity. Thus, developmental acceleration allows spadefoot toad tadpoles to evade drying ponds, but it comes at the expense of reduced size at metamorphosis and increased oxidative stress.     

Oviposition site selection by mosquitoes is affected by cues from conspecific larvae and anuran tadpoles

Abstract The oviposition site that a female mosquito selects will influence the fitness of her larvae. We conducted a series of artificial pond experiments to compare the oviposition responses of two species of mosquitoes with the presence of tadpoles, conspecifics and chemical cues from these organisms. The two mosquito species differ markedly in larval ecology. The larvae of one species, Culex quinquefasciatus, co‐occur with numerous freshwater organisms, including tadpoles of Linmodynastes peronii (the striped marsh frog). Larvae of the other mosquito, Ochlerotatus australis, inhabit small brackish rock ponds where the main potential competitors are tadpoles of Crinia signifera (the common eastern froglet). In field trials, females of both mosquito species oviposited significantly more often in water that contained (or had previously contained) conspecific larvae. However, these superficially similar responses were mediated via different pathways: fungicide abolished the response by C. quinquefasciatus but not by O. australis. The two mosquito species also responded differently to cues associated with syntopic tadpoles. The presence of tadpoles did not influence oviposition by C. quinquefasciatus, but O. australis oviposited less often if tadpoles were present. These interspecific differences in oviposition behaviour may be adaptive to differences in larval ecology: competition with tadpoles is likely to be more significant for O. australis than for C. quinquefasciatus. Our findings thus support the hypothesis that mosquitoes oviposit selectively to avoid potential anuran larval competitors.     

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.     

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.     

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.     

Effect of changes in resource level on age and size at metamorphosis in Hyla squirella

Recent experiments suggest that timing of metamorphosis is fixed during development in some anurans, insects, and freshwater invertebrates. Yet, these experiments do not exclude a growth rate optimization model for the timing of metamorphosis. I manipulated food resources available to larvae of squirrel treefrogs (Hyla squirella) to determine if there is a loss of plasticity in duration of larval period during development and to critically test growth rate models for the timing of metamorphosis. Size-specific resource levels for individual tadpoles were switched from low to high or high to low at three developmental stages spaced throughout larval development. The effects of changes in resource availability on larval period and mass at metamorphosis were measured. Switching food levels after late limb bud development did not significantly affect larval period in comparison to constant food level treatments. Therefore, developmental rate in H. squirella is better described by a fixed developmental rate model, rather than a growth rate optimization model. The timing of fixation of developmental rate in H. squirella is similar to that found in other anuran species, suggesting a taxonomically widespread developmental constraint on the plasticity of larval period duration. Mass at metamorphosis was not significantly affected by the timing of changes in food levels; the amount of food available later in development determined the size at metamorphosis. Larval period and mass at metamorphosis were negatively correlated in only one of two experiments, which contrasts with the common assumption of a phenotypic trade-off between decreased larval period and increased mass at metamorphosis. Received: 19 August 1996 / Accepted: 20 June 1997     

Effects of light intensity on activity in four sympatric anuran tadpoles

Though light conditions are known to affect the development and anti-predation strategies of several aquatic species, relatively little is known about how different species react to light, or how light can affect these species during different points in their life-cycle. In this study, we used four sympatric anuran tadpoles(Bufo gargarizans, B. melanostictus, Pelophylax nigromaculatus and Microhyla fissipes) as animal system to examine species-specific activities of the underdoing different light intensity treatments, so as to better understand how they respond to light. We exposed four different species of tadpoles to 1660 and 14 lux light intensity treatments and then measured several parameters including development stage, body length and tail length, and as well as their basic activities. The results of this observation and analysis showed that the activities of tadpoles were significantly greater in B. gargarizans and B. melanostictus than in P. nigromaculatus and M. fissipes; and were also significantly greater during times of high light intensity as compared to during low light intensity. Moreover, the observed relationship between species and light intensity was significant. The activities of B. gargarizans and B. melanostictus tadpoles were greater in high light, while the activity of P. nigromaculatus tadpoles was greater in low light intensity, while M. fissipes tadpoles showed no differences in either low or high intensity light. Furthermore, the activities of B. gargarizans, B. melanostictus and M. fissipes tadpoles in terms of developmental stage, body size or tail length did not seem to differ with light intensity, but during early larval developmental period of P. nigromaculatus, the activity of tadpoles was negatively correlated with development stage, but irrelevant to either body size or tail length in different light intensities. These results lead us to conclude the observed activities of the four sympatric anuran tadpoles are closely correlated with their specific anti-predation strategies.     

外来捕食者克氏原螯虾对黑斑侧褶蛙不同发育阶段幼体表型与运动能力的影响

外来物种入侵是造成本土无尾两栖类种群下降和灭绝的最严重威胁之一。在中国南部丽水地区,克氏原螯虾(Procambarus clarkiion)是一种捕食本土无尾类蝌蚪的入侵物种。在实验室条件下,通过对黑斑侧褶蛙(Pelophylax nigromaculatus)受精卵和蝌蚪先后进行克氏原螯虾捕食风险的不同处理,检测孵出蝌蚪的形态特征、中期蝌蚪的形态特征、游泳表现及活动水平、变态时间与大小以及幼蛙跳跃表现,评估克氏原螯虾的捕食风险对黑斑侧褶蛙不同生长阶段幼体生长发育、运动能力及行为的综合影响。单因素方差分析表明,捕食者长期存在条件下黑斑侧褶蛙孵出蝌蚪的体长、尾长和尾肌宽均显著短于无捕食者的空白对照组和捕食者仅存在2 d的实验组,后两者处理下的蝌蚪体长差异显著,但尾长和尾肌宽差异不显著。长期捕食者存在条件下黑斑侧褶蛙蝌蚪的游泳总路程和平均游速均显著小于无捕食者和捕食者存在2 d的实验组,而蝌蚪体长、尾长、尾鳍高和尾肌高的组间差异均不显著。有捕食者存在下黑斑侧褶幼蛙的体长和体重均显著大于无捕食者和捕食者存在2 d的实验组的情况,而变态时间和幼蛙跳跃距离组间差异不显著。两因素方差分析表明,测试时间、捕食者设置和两因素的相互作用均显著影响蝌蚪的活动频次。不同实验时间,长期捕食者存在实验组蝌蚪活动频次均小于无捕食者和捕食者存在2 d的实验组。无捕食者实验组蝌蚪的活动频次前期高于捕食者存在2 d的实验组,后期低于捕食者存在2 d的实验组。线性回归分析结果显示,捕食者存在2 d实验组蝌蚪相对活动频次与检测时间相关不显著,长期捕食者存在实验组相对蝌蚪活动频次与检测时间呈显著的正相关。本研究表明了黑斑侧褶蛙蝌蚪能够感知来自入侵捕食者克氏原螯虾的捕食风险,诱导产生较小的孵出个体、蝌蚪较低的活动水平,但这种不足可通过后期补偿生长,不影响进入陆地生境的变态个体的表型与运动能力。     

Sequence of ontogenetic development and atrophy of the oral apparatus of six anuran tadpoles

The development of the oral structures of six species of anuran tadpoles with four different types of mouth parts and the metamorphic atrophy of these structures in two species with different mouth parts are described. The oral labia of typical tadpoles, oral flaps of microhylids, and lateral oral folds of Rhinophrynus are assumed to be homologous. We also suggest that the barbels of the tadpoles of Rhinophrynus are homologs of the marginal papillae of species with an oral disc. Developmental patterns and sequences of the oral structures of all tadpoles examined follow a common pattern: stomodeal invagination, oral pad development, jaw sheeth delimitation, tooth row ridge development, jaw sheath keratinization, and labial tooth keratinization. Developmental patterns remain constant, while interspecific differences are apparent because of truncations of ontogeny at specific stages. Metamorphic atrophy of oral structures occurs roughly in the reverse order of development, although the procedure is rapid and more haphazard than development.     

Risk of predation and behavioural response in three anuran species: influence of tadpole size and predator type

Many species alter their activity, microhabitat use, morphology and life history in response to predators. Predation risk is related to predator size and palatability of prey among others factors. We analyzed the predation risk of three species of tadpoles that occur in norwestern Patagonia, Argentina: Pleurodema thaul, Pleurodema bufoninum and Rhinella spinulosa. We sampled aquatic insect predators in 18 ponds to determine predator–tadpole assemblage in the study area. In laboratory conditions, we analysed the predation rate imposed by each predator on each tadpole species at different tadpole sizes. Finally, we tested whether tadpoles alter their activity in the presence of chemical and visual cues from predators. Small P. thaul and P. bufoninum tadpoles were the most vulnerable prey species, while small R. spinulosa tadpoles were only consumed by water bugs. Dragonflies and water bugs were the most dangerous tadpole predators. Small P. thaul tadpoles reduced their activity when they were exposed to all predators, while large tadpoles only reduced the activity in the presence of large predators (dragonfly larvae and water bugs). Small P. bufoninum tadpoles reduced the activity when they were exposed to beetle larvae and dragonfly larvae, while large tadpoles only reduced activity when they were exposed to larger predators (water bugs and dragonfly larvae). R. spinulosa tadpoles were the less sensitive to presence of predators, only larger tadpoles responded significantly to dragonfly larvae by reducing their activity. We conclude that behavioural responses of these anuran species were predator-specific and related to the risk imposed by each predator.     

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     

Factors determining tadpole vulnerability to predators: can prior experience compensate for a suboptimal shape?

We investigated the role of constitutive morphology and previous experience in predator avoidance in two anuran species associated with different larval habitats. In Rana temporaria, deeper tails and larger body size conferred selective advantage against dragonfly predation. Previous experience with predators had a positive influence on the survival of R. temporaria tadpoles equivalent to predator selection. By contrast, survival in Bufo bufo seems unrelated to tail shape or experience. This suggests that B. bufo lacks constitutive morphological defenses against insect predators, and that morphological and behavioral defenses could result more effective than chemical deterrents for these insect predators. A key novelty of this study is the observation that Rana tadpoles having prior experience with predators have an enhanced success in further encounters, and this occurs before the morphological induced defense has been established. This induced modification for R. temporaria, and its lack of for B. bufo, may be an important determinant of larval survival.     

The ontogeny of Pseudis platensis (Anura,Hylidae): Heterochrony and the effects of larval development on postmetamorphic life

Recent studies have described the giant tadpole, delayed metamorphic transformations, and absence of postmetamorphic growth of the skeleton of Pseudis Platensis. These features address questions about derived patterns of life cycles and the role of the heterochrony during the metamorphosis in anurans. Using anatomical methods, we provide new data on the development of reproductive, digestive and integument systems, and age inference obtained from ontogenetic series of Pseudis platensis. Our results indicate that at the end of metamorphosis, the adult skin is completely differentiated, including the calcified dermal layer; the testis has seminiferous tubules with spermatogonia, spermatocytes, and spermatids; ovarian sacs present previtellogenic ova; and the adult digestive tract is fully formed. The froglets differ from adults only in being unable to reproduce. The entire life cycle of P. platensis can occur in 4 years. In the first year, larval development, growth to adult size, and gonad differentiation are completed. Long larval development rather than size of the tadpoles seems to be involved in the absence of juvenile stages. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Discovery of a Novel Alveolate Pathogen Affecting Southern Leopard Frogs in Georgia: Description of the Disease and Host Effects

In April of 2006, we observed southern leopard frog (Rana sphenocephala) tadpoles in a pond in northeast Georgia that were dying from an unknown pathogen. Examination of affected specimens, as well as PCR characterization, revealed that all were infected with a novel alveolate pathogen closely related to freshwater and marine eukaryotic organisms and, to a lesser degree, to members of the genus Perkinsus. This pathogen has been documented in numerous mortality events in anuran tadpoles in the United States, although it has not yet been named nor clearly described. We subsequently conducted a systematic survey of this and four other ponds in the same area to document the extent of the pathogen and to describe the nature of infections in leopard frog tadpoles. Of 87 live tadpoles examined, 25% were infected with the alveolate pathogen, based on visual inspection of tadpole liver tissue. Affected tadpoles frequently had enlarged abdomens, swam erratically, and could be captured by hand. All organs of infected tadpoles were infiltrated but typically to a lesser extent than the liver and kidneys, which often had hundreds of thousands of the spherical, 6-μm organisms. Infected tadpoles tended to weigh more than noninfected ones, likely due to the massive organ swelling that coincided with infections. Infected tadpoles did not differ in developmental stage from noninfected tadpoles. Infection prevalence varied widely among ponds, and in one pond, we witnessed a rapid die-off of R. spenocephala tadpoles during our surveys, although we did find infected metamorphic frogs. The rapid mortality we observed as well as the vast number of organisms seen in specimens suggests that this pathogen has tremendous transmission potential, and therefore deserves further monitoring and study.