Tadpole Body Research Articles

Reproductive ecology of Sichuan digging frogs (Microhylidae: Kaloula rugifera )

Abstract. We investigated the reproductive ecology of Sichuan digging frogs (Microhylidae: Kaloula rugifera ) in Mianyang, China during the wet season of 2013. Male Sichuan digging frogs first appear at temporary ponds following the first heavy rain of the wet season and initiate calling. Females arrive at ponds shortly after males. Male frogs chorus extensively throughout the wet season during the evenings and nights following rainstorms. Female frogs leave the pond after laying eggs, and likely only lay one clutch annually. Amplexus lasted up to three hours. Females were larger than males in terms of body size, but we found no evidence of size-assortative mating. Clutch size varied from 920 to 2200 eggs, with egg diameter ranging from 1.33 to 1.93 mm. Larger female frogs laid more eggs, and there was no correlation between egg number and egg size. Tadpoles hatched from eggs within 18-20 hours of oviposition, and grew for 30-40 days before complete metamorphosis occurred. The initial body length of tadpoles ranged from 3-5 mm snout-vent length. Growth was fastest immediately after hatching, and declined asymptotically with increasing tadpole body size. Overall, Sichuan digging frogs have a breeding biology characterized by strong male-male competition with prolonged breeding coinciding with the annual wet season. Key w ords . Breeding ecology; Kaloula rugifera ; life history; mating system

Acute toxicity and sublethal effects of the mixture glyphosate (Roundup Active) and Cosmo-Flux 411F to anuran embryos and tadpoles of four Colombian species.

Glyphosate is the most widely used herbicide in the world with application in agriculture, forestry, industrial weed control, garden and aquatic environments. However, its use is highly controversial for the possible impact on not-target organisms, such as amphibians, which are vanishing at an alarming and rapid rate. Due to the high solubility in water and ionic nature, the glyphosate requires of surfactants to increase activity. In addition, for the control of coca (Erythroxylum coca) and agricultural weeds in Colombia, formulated glyphosate is mixed and sprayed with the adjuvant Cosmo-Flux 411F to increase the penetration and activity of the herbicide. This study evaluates the acute toxic and sublethal effects (embryonic development, tadpole body size, tadpole swimming performance) of the mixture of the formulated glyphosate Roundup Active and Cosmo-Flux 411F to anuran embryos and tadpoles of four Colombian species under 96h laboratory standard tests and microcosms, which are more similar to field conditions as they include soil, sand and macrophytes. In the laboratory, embryos and tadpoles of Engystomops pustulosus were the most tolerant (LC50 = 3904 microg a.e./L; LC50=2 799 pg a.e./L, respectively), while embryos and tadpoles of Hypsiboas crepitans (LC50=2 203 microg a.e./L; LC50=1424 microgg a.e./L, respectively) were the most sensitive. R. humboldti and R. marina presented an intermediate toxicity. Embryos were significantly more tolerant to the mixture than tadpoles, which could be likely attributed to the exclusion of chemicals by the embryonic membranes and the lack of organs, such as gills, which are sensitive to surfactants. Sublethal effects were observed for the tadpole body size, but not for the embryonic development and tadpole swimming performance. In microcosms, no toxicity (LC50 could not be estimated), or sublethal responses were observed at concentrations up to fourfold (14.76 kg glyphosate a.e./ha) the highest field application rate of 3.69 kg glyphosate a.e./ha. Thus, toxicity was less in the microcosms than in laboratory tests, which may be attributed to the presence of sediments and organic matter which rapidly adsorb glyphosate and surfactants such as POEA. It is concluded that the mixture of glyphosate (Roundup Active) and Cosmo-Flux*411F, as used in the field, has a negligible toxic effect to embryos and tadpoles of the species tested in this study.

Predator‐induced defenses in tadpoles confound body stoichiometry predictions of the general stress paradigm

Predation is known to have both direct and indirect effects on nutrient cycling in terrestrial and aquatic ecosystems, and the general stress paradigm (GSP) has been promoted as a theory for describing predator-mediated indirect effects on nutrient cycling. The GSP predicts that prey exposed to predators will produce glucocorticosteroids, which have a host of physiological effects including gluconeogenesis, increased respiration, excretion of N and P, and increases in body C:N. We tested the nutrient predictions of the GSP using anuran larvae, which exhibit morphological defenses in addition to behavioral defenses for which the GSP was conceived. Genetically similar Hyla versicolor tadpoles were placed in mesocosms either in the presence or absence of a fed predator (Dytiscus verticalis), and after two weeks, tadpoles exposed to predators exhibited strong induced defenses with large, tubular bodies, larger tails, and reduced activity. Tadpole body %C and N:P increased with no change in C:N, which is contrary to expectations from the GSP. Statistical models suggested that changes in body morphology (e.g., tail muscle width) rather than behavioral defenses (i.e., reduced activity) were most likely responsible for predator-mediated differences in body stoichiometry. This study suggests that strong morphological defenses may overwhelm or counteract the nutrient predictions of the GSP.

Expression analysis of XPhyH-like during development and tail regeneration in Xenopus tadpoles: Possible role of XPhyH-like expressing immune cells in impaired tail regenerative ability

Xenopus tadpoles have high regenerative ability of amputated tails except during the ‘refractory period’, when the ability is transiently lost. We previously demonstrated that distinct immune responses occur in tail stumps between the refractory and pre/post-refractory regeneration periods. Furthermore, treatment with an immunosuppressant, FK506, restores the tail regenerative ability during the refractory period. Based on these findings, we previously proposed that autoreactive immune cells infiltrate the tail stumps to attack blastema cells as ‘non-self’ during the refractory period, resulting in the impaired regenerative ability. The immune cells that attack the blastema cells, however, remained unclear. Here we screened for genes whose expression in the tail stumps was altered by FK506 treatment during the refractory period and identified a Xenopus homolog of phytanoyl-CoA dioxygenase (PhyH)-like. XPhyH-like expression transiently increased in tail stumps after amputation during the refractory period, and was reduced by FK506 treatment. XPhyH-like expression in the whole tadpole body specifically increased during the refractory period and was enriched in the blood cell fraction. These findings suggest that XPhyH-like is expressed in autoreactive immune cells that are distributed in the whole body during the refractory period and transiently infiltrate the tail stumps to attack the blastema cells as ‘non-self’.

Effect of titanium dioxide nanomaterials and ultraviolet light coexposure on African clawed frogs (Xenopus laevis)

Titanium dioxide nanomaterials (nano-TiO(2) ) exhibit stronger photochemical oxidation/reduction capacity compared with their bulk counterparts, but the effectiveness of nano-TiO(2) interaction with ultraviolet (UV) light strongly depends on particle size. In this study, the dependence of nano-TiO(2) toxicity on particle size and interaction with UV light were investigated. Toxicity tests with Xenopus laevis included eight concentrations of nano-TiO(2) in the presence of either white light or UVA (315-400 nm). We quantified viability and growth of Xenopus laevis. Results showed that, regardless of UV light exposure, increasing TiO(2) concentration decreased X. laevis survival (p < 0.05). Coexposure to 5-nm TiO(2) and UVA caused near-significant decreases in X. laevis survival (p = 0.08). Coexposure to 10-nm TiO(2) and UVA significantly decreased X. laevis survival (p = 0.005). However, coexposure to 32-nm TiO(2) and UVA had no statistical effect on X. laevis survival (p = 0.8). For all three particle sizes, whether alone or with UV light, the nano-TiO(2) concentrations significantly affected growth of tadpoles as determined by total body length, snout-vent length, and developmental stage. High-concentration TiO(2) solutions suppressed tadpole body length and delayed developmental stages. Further research to explore reasons for the growth and mortality in tadpoles is still underway in our laboratory. Given the widespread application of nano-TiO(2) , our results may be useful in the management of nano-TiO(2) released from industrial, municipal, and nonpoint sources.

Grazing on toxic cyanobacterial blooms by tadpoles of edible frog Rana grylio

SUMMARYTadpoles of Rana grylio were raised as edible frogs in fishponds of Guanqiao in Wuhan City, Hubei, China, during cyanobacterial blooms from June to October. The dominant cyanobacterial species was Microcystis, which was found to be lethally toxic by intraperitoneal (i.p.) mouse bioassay. Little is known about the effect of tadpoles on toxic cyanobacterial blooms. To evaluate the potential of the tadpoles to graze on cyanobacterial blooms, the tadpoles were fed on Microcystis collected from the field in the laboratory. The Microcystis cells decreased from 1.19 × 107 cells mL−1 to 3.23 × 106 cells mL−1, with a sharp reduction of 73% of the initial Microcystis population observed in the first 24 h after introduction of the tadpoles. The ponds containing tadpoles had a markedly lower density of Microcystis than those lacking tadpoles. Tadpoles exposed to either cultured Microcystis aeruginosa (NIES–90, 2.768 µg microcystins mg–1 dw–1) cells or lysed M. aeruginosa cells grew well, however, indicating that they were unaffected by Microcystis toxins. We found a significant increase in tadpole body weight after feeding on either field Microcystis or cultured M. aeruginosa. The mean increase in individual body weight was 20 mg day−1 when fed on Microcystis from the pond, and 7 mg day−1 when fed on M. aeruginosa from culture. Our study strongly suggested that there is a direct trophic relationship between R. grylio tadpoles and toxic Microcystis blooms and they possess the potential to graze on toxic Microcystis. The results imply that R. grylio tadpoles may play an important ecological role in reducing toxic cyanobacterial blooms caused by Microcystis.

Larval alarm pheromones as a potential control for invasive cane toads (Bufo marinus) in tropical Australia

Novel approaches to control invasive species are urgently needed. Cane toads (Bufo marinus) are large, highly toxic anurans that are spreading rapidly through tropical Australia. Injured toad larvae produce an alarm pheromone that elicits rapid avoidance by conspecifics but not by frog larvae. Experiments in outdoor ponds show that repeated exposure to the pheromone reduced toad tadpole survival rates (by >50%) and body mass at metamorphosis (by 20%). The alarm pheromone did not induce tadpoles to seek shelter, but accelerated ontogenetic differentiation. Perhaps reflecting mortality of weaker individuals during larval life, growth rates post-metamorphosis were higher in animals emerging from the pheromone exposure treatment than from the control treatment. Nonetheless, body size differentials established at metamorphosis persisted through the first 8 days of post-metamorphic life. We will need substantial additional research before evaluating whether the alarm pheromone provides a way to reduce cane toad recruitment in nature, but our field trials are encouraging in this respect.

Vulnerability of an Australian anuran tadpole assemblage to the toxic eggs of the invasive cane toad (Bufo marinus)

The invasion of cane toads (Bufo marinus) across tropical Australia has fatally poisoned many native predators; the most frequent victims may be tadpoles of native frogs, which die when they consume the toxic eggs of the toads. Field studies have documented high and species-specific mortality of tadpoles following toad spawning.To clarify the determinants of tadpole vulnerability, we conducted 1593 laboratory trials in which single tadpoles were exposed to 10 toad eggs, either with or without an alternative food source (lettuce). At least some tadpoles within all 15 species tested consumed toad eggs. Interspecific variance in survival rates (from 0 to >70%) was driven by feeding responses not by physiological tolerance to toxins: almost all native tadpoles that consumed eggs died rapidly.Tadpole mortality was decreased by the presence of an alternative food source in four species, increased in two species, and not affected in seven species. In three of four taxa where we tested both small (early-stage) and large (late-stage) tadpoles, both mean survival rates and the effects of alternative food on survival shifted with tadpole body size. Trials with one species (Limnodynastes convexiusculus) showed no significant inter-clutch variation in feeding responses or tolerance to toxins. Overall, our data show that cane toad eggs are highly toxic to native anuran tadpoles, but that whether or not a tadpole is killed by encountering toad eggs depends upon a complex interaction between the native anuran's species, its body size, and whether or not alternative food was present. In nature, larval vulnerability also depends upon the seasonal timing and location of spawning events, and habitat selection and foraging patterns of the tadpoles. Our results highlight the complexity of vulnerability determinants, and identify ecological factors (rather than physiology or feeding behaviour) as the primary deter- minants of cane toad impact on native tadpoles.

Reproductive Biology, Vocalizations, and Tadpole Morphology ofRupirana Cardosoi, an Anuran with Uncertain Affinities

ABSTRACT. We describe the tadpole morphology, the advertisement and courtship calls, and reproductive characteristics of Rupirana cardosoi Heyer 1999, an endemic species from rocky mountain fields in Chapada Diamantina, Bahia State, Brazil. Females of Rupirana cardosoi deposit pigmented eggs in shallow backwaters or ponds. The body of tadpoles is ovoid in dorsal and ventral views and depressed in lateral view. The oral disc is ventral and the labial tooth row formula is 2(2)/3(1). Eyes are dorsal. The advertisement call has one pulsed note, with dominant frequency approximately 1.3 kHz. The courtship call presents three note types with dominant frequency ranging from 0.7 to 1.4 kHz. Females are larger ( = 34.4, SD = 1.6, 31.4–37.4 mm, n = 12) than males ( = 30.9, SD = 1.5, 28.8–33.8, n = 9; t = 5.1; p < 0.001) and males present nuptial excrescences located on the base of the thumbs. The eggs are medium sized ( = 2.47 ± 0.52 mm, N = 27) and mean clutch size was 120.44 ± 74.38 eggs (N = 11 clutches).

Giant Tadpole and Delayed Metamorphosis of Pseudis platensis Gallardo, 1961 (Anura, Hylidae)

Pseudis platensis, like Pseudis paradoxa, is a Neotropical aquatic frog with a giant tadpole. Despite being one of the few anuran species whose larvae grow larger than adults, knowledge of its larval development is limited to prometamorphic stages. This study describes qualitative and quantitative variation through P. platensis larval development, including descriptions of various traits (lateral line, oral disc, vent tube, limbs) and their transformations during metamorphosis. We found similarities with the development of other anurans at larval stages up to the end of prometamorphosis but not with their metamorphic stages. For this species, we described key features to characterize 13 ontogenetic stages as follows: four for prometamorphic larval morphologies, eight stages covering metamorphic changes, and one defining the posmetamorphic adult stage. At the end of prometamorphosis, tadpole body size in P. platensis is similar to adult size, but the tail shows extra development. Unlike most anurans, the disappearance of two larval traits (i.e., vent tube and tail) is delayed until later in metamorphosis. Larval development of P. platensis exhibits unique features not reported for other anurans and offers a model to study developmental and evolutionary aspects of anuran metamorphosis.

Getting in shape: adaptation and phylogenetic inertia in morphology of Australian anuran larvae

The external morphology of frog larvae is predicted to vary among habitats, for a variety of functional reasons. I performed a phylogenetic comparative study to test whether correlations between habitat and the shape of the tadpole and its oral disc are adaptive in 82 species from south-eastern Australia in the families Hylidae and Myobatrachidae. Habitat distributions and phylogenetic relationships were compiled from the literature and shape was quantified using geometric morphometric analysis of published drawings. Results indicate that shape evolved towards different optima in different habitats while also showing appreciable levels of phylogenetic inertia. Within myobatrachids, evolution of terrestrial tadpoles was associated with a short and shallow head/body and a shallow tail. In aquatic species, the use of benthic microhabitats was correlated with a long shallow tail, dorsal eye position, shallow head/body and ventral mouth with robust jaw sheaths. Even traits with evidence for adaptation evolved slowly in response to habitat shifts, usually requiring >or=10 million years to evolve half-way to a new optimum. Although these findings support adaptive evolution of tadpole body form, they also highlight a strong influence of ancestral character states and indicate that phenotypes in extant species are partly maladaptive.

Factors Influencing Responses to Alarm Pheromone by Larvae of Invasive Cane Toads, Bufo marinus

If pheromonal communication systems of invasive species differ from those of native biota, it may be possible to control the invader by exploiting that difference. When injured, the larvae of cane toads, Bufo marinus, an invasive species of major concern in tropical Australia, produce species-specific chemical cues that alert conspecific tadpoles to danger. Repeated exposure to the alarm chemical reduces tadpole survival rates and body sizes at metamorphosis and, thus, could help control toad populations. To evaluate the feasibility of this approach, we need to know how the intensity of toad tadpole response to the alarm chemical is affected by factors such as water temperature, time of day, larval stage and feeding history, geographic origin of the tadpoles, and habituation. Information on these topics may enable us to optimize deployment, so that tadpoles encounter pheromone at the times and places that confer maximum effect. In our studies, tadpole density, nutritional state, larval stage, and geographic origin had little effect on the intensity of the alarm response, but tadpoles reacted most strongly in higher water temperatures and during daylight hours. Repeated, once-daily exposure to pheromone did not induce habituation, but repeated exposure at 15-min intervals did not elicit further responses after 2 h total exposure. The insensitivity of response to most factors tested means that the effectiveness of the pheromone as a control agent should be relatively robust.

The immune system is involved in Xenopus metamorphosis

Amphibian metamorphosis provides a model to elucidate the mechanisms underlying how vertebrates reconstitute a body plan and how the immune system develops during ontogeny. In Xenopus, T cells are expanded from the early developmental stages just after hatching. These T cells switch from larval-type in an easily tolerizable state into an adult-type having a potent immune responsiveness comparable to that of mammals. During metamorphosis, tadpoles exhibit morphological changes in skin that completely transforms from larval-type to adult-type. Only tail tissue behaves differently; it remains a larval-type tissue until it disappears at the end of metamorphosis. Thus, at metamorphic climax, four different types of cells co-exist in a tadpole body: larval tissue cells; adult tissue cells; larval immune cells; and adult immune cells. Based on the results showing that tadpole tail skin is rejected by syngeneic adult, it is proposed that the elimination of the larval tissue cells by the adult T cells that occurs during metamorphosis is immunologically mediated. Recent results indicate that the antigenic proteins expressed in the metamorphosing skin cells participate in the process of tail regression. This chapter describes how animals adjust and survive through such crises associated with large scale replacement of entire body cells.

Anatomy of anuran tadpoles from lentic water bodies: systematic relevance and correlation with feeding habits

I studied anatomy, gut content, and the relationship among these traits in a set of anuran tadpoles. Larval stages (mainly Gosner stages 31–36) of nineteen species from various lentic environments were selected. Morphological characters from the skeleton, musculature, oral apparatus and buccopharyngeal cavity were recorded, and a gut content analysis was performed, with emphasis on food size distribution. Ordination techniques were applied in order to find patterns of similarity in morphology and gut content. Canonical ordination methods were used to investigate the relationship among gut content, morphology, and phylogeny in the species considered. The results show that several skeletal, muscular, and buccal characters are relatively maintained within genera. Other features, which have appeared independently in different lineages, reflect convergence phenomena in some cases related to ecological aspects. The configuration of the hyobranchial skeleton, the development of the buccal floor depressor and levator muscles, and mouth gape width correlate with prey size. In some species, morphology is clearly related with feeding. Tadpoles that ingest large food particles relative to their body length present morphological traits attributable to macrophagy. Taxonomically unrelated tadpoles of Dendropsophus nanus, D. microcephalus and Ceratophrys cranwelli possess hyobranchial skeletons with robust, rostrocaudally long ceratohyals and reduced branchial baskets with short ceratobranchials devoid of lateral projections and spicules. Lepidobatrachus llanensis tadpoles have laterally extended ceratohyals which, along with the lateral extension of the jaws, result in a very wide oral apparatus and an ample buccopharyngeal cavity that allows the tadpole to ingest large and whole prey; the branchial basket, although its ceratobranchials lack lateral projections and spicules, is slightly reduced in area. The four species mentioned have a noticeable development of the buccal floor depressor muscles, and buccal cavities with scarce filtering and entrapping structures. In Elachistocleis bicolor, Dermatonotus muelleri, Chiasmocleis panamensis, and Xenopus laevis tadpoles, the branchial basket occupies &gt;70% of the total hyobranchial skeleton area, and the hypobranchial plates are highly reduced; the buccal floor levator muscles are well-developed, with an increased site of attachment on the ventral expansion of the lateral process of the ceratohyal; the scarcity of the filtering structures in the buccopharyngeal cavity are balanced with the great development of the branchial filters and secretory zones; all these features relate to a diet based on small particles not significantly different from those of most other species; however, experimental studies show that species with similar hyobranchial apparatus and muscles are the most efficient when retaining minute particles. Finally, a large group of species present generalized morphological characters, such as a branchial basket occupying about 50% of the total hyobranchial apparatus, intermediate values of mouth gape width and buccal floor levator / depressor muscles ratio, and abundant filtering structures in the buccopharyngeal cavity; these species feed frequently on food particles between 1–30% of the tadpole body length; however, in some of the species, macrophagous diets are also reported in the literature, indicating that this morphology is flexible in more ample prey size ranges.

Exposures to Estradiol, Ethinylestradiol and Octylphenol Affect Survival and Growth of Rana pipiens and Rana sylvatica Tadpoles

Endocrine disrupting compounds (EDCs) are often detected in the aquatic environment and can negatively affect the health of wildlife populations. However, little is known about the sensitivity of native amphibians to EDCs. Wood frogs (Rana sylvatica) and Northern leopard frogs (Rana pipiens) were exposed to three estrogenic EDCs: estradiol (E2), ethinylestradiol (EE2), and 4-tert-octylphenol (OP). In addition, R. pipiens were exposed during two developmental stages (Gosner stages 26 and 36) to examine life-stage differences in sensitivity. Tadpoles were exposed for 2 wk to 8 nominal concentrations (0.25 μM–10 μM) of each compound. Individual mortality was recorded during the exposure period, while body weight was measured at the end of 2 wk. LC50 values were calculated, and differences in body weight between vehicle control and exposed groups were assessed. Rank order toxicity of the compounds for both R. pipiens stages and both species was OP > EE2 > E2. Gosner stage 26 tadpoles were more sensitive (LC50: E2 [5.57 μM], EE2 [3.01 μM], OP [1.36 μM]) to all three compounds when compared to stage 36 tadpoles (LC50: E2 [>10 μM], EE2 [4.17 μM], OP [2.80 μM]). Interspecies comparisons revealed R. sylvatica tadpoles (LC50: E2 [2.50 μM], EE2 [1.89 μM], OP [0.74 μM]) as being more sensitive to the three compounds than R. pipiens (LC50: E2 [4.56 μM], EE2 [2.75 μM], OP [1.42 μM]). Xenoestrogen exposure also affected tadpole body weight which may have long-term adverse effects on the rate of metamorphosis. These results provide toxicological data needed for assessing sublethal effects of estrogenic compounds on amphibian development and suggest that environmental levels of OP may pose a serious risk to the health of amphibian populations.

Tadpole Body Zones Differ with Regard to Strike Frequencies and Kill Rates by Dragonfly Naiads

We assessed Anax junius dragonfly naiad strike frequencies and kill probabilities against Rana sylvatica (Wood Frog) tadpoles. Strikes fell into five categories according to their placement on the tadpole body: anterior head/body, posterior head/body, head/body-tail intersection, anterior tail, and posterior tail. The kill probability was calculated for each category as the number of kills divided by the number of contacts made in that category. These probabilities varied; a higher probability was found for both the anterior and posterior head/body, and values declined for successively posterior categories. Neither the kill probability nor the size of the body zone influenced the number of strikes to that zone. Surprisingly, the dragonfly naiads struck most frequently at the relatively narrow region of the head/body-tail intersection, suggesting that dragonfly naiads may aim at a specific target.

Genetic basis of phenotypic plasticity for predator-induced morphological defenses in anuran tadpole, Rana pirica, using cDNA subtraction and microarray analysis

Anuran tadpoles ( Rana pirica) are induced to develop a higher tail and a bulgy body as predator-specific morphological responses when they are exposed to predatory larval salamanders. Subtractive hybridization was performed using induced tadpole body skin and normal tadpoles’ body skin. A total of 196 clones showed higher expression, and 104 clones showed lower expression, when they formed bulgy bodies. In the subtraction, carboxypeptidase B, trypsinogen, elastase I, fibrinogen, elastase II, triacyl-glycerol lipase, and α1-antitrypsin genes showed lower expression. In contrast, RT-like protein, bullous pemphigoid antigen, phosphoserine aminotransferase, uromodulin, tetranectin, chaperonin-like protein, zinc finger protein, osteonectin, aldehyde dehydrogenase, Sec 23A protein, and ribosomal protein showed higher gene expression. Microarray analysis was also performed using this subtracted cDNA (nine replicates). Results of the microarray data essentially corresponded with those of the subtraction data, and the degree of the suppressed genes was much stronger than that of the expressed genes. Carboxypeptidase B showed the strongest suppression, and its inhibition range was from 1/100 to 3/100 compared with that of control body skin. Strong suppression was also observed with trypsinogen, elastase I, fibrinogen, and elastase II as well. These results can be interpreted as increases of fibrinolysis by strong depression of both carboxypeptidase B and other genes simultaneously, resulting in the retention of blood vessels and facilitating the circulation of blood. Expression was observed in phosphoserine aminotransferase, aldehyde dehydrogenase, RT-related protein, chaperonin-like protein, tetranectin, bullous pemphigoid antigen, uromodulin, and Sec 23A protein. They were significantly ( p < 0.05) increased and were at least 1.5 times greater compared with the control. From the appearance, it seems that the bulgy shaped body is highly connecting to the bullous pemphigoid (BP) antigen that causes the skin blistering disorder, and tetranectin and uromodulin may be related to the extracell matrix through myogenesis, protein secretion, and ion transport, respectively. Since the RT-related protein gene derived from retrotransposon (L1) is known to disrupt mammalian transcriptomes, retrotransposon may be involved with phenotypic plasticity for morphological defense by Rana prica against predator threat.

A gynogenetic screen to isolate naturally occurring recessive mutations in Xenopus tropicalis

In the rapidly developing, diploid amphibian Xenopus tropicalis, genetics can be married to the already powerful tools of the amphibian system to overcome a disability that has hampered Xenopus laevis as a model organism: the difficulties inherent in conducting genetic analyses in a tetraploid organism with a longer generation time. We describe here a gynogenetic screen to uncover naturally occurring recessive mutations in wild X. tropicalis populations, a procedure that is both faster and easier than conventional genetic screens traditionally employed in model organisms to dissect early developmental pathways. During the first round of our screen, gynogenetic diploids from over 160 females comprising four different wild-caught populations were examined. Forty-two potential mutant phenotypes were isolated during this round of gynogenesis. From this group, we describe 10 lines that have genetically heritable recessive mutations. A wide range of developmental defects were obtained in this screen, encompassing effects limited to individual organs as well phenotypes characterized by more global changes in tadpole body morphology. The frequency of recessive mutations detected in our screen appears lower than that seen in other vertebrate genetic screens, but given constraints on the screening procedure used here, is likely to be consistent with rates seen in other animals, and clearly illustrates how wild-caught animals can be a productive source of developmental mutations for experimental study. The development of genetic strategies for the Xenopus system, together with new genomic resources, existing technologies for transgenesis, and other means for manipulating gene expression, as well as the power of performing embryonic manipulations, will provide an impressive set of tools for resolving complex cell and developmental phenomena in the future.

On the behavioural response ofRanaandBufotadpoles to echinostomatoid cercariae: implications to synergistic factors influencing trematode infections in anurans

We used high-speed videography of staged encounters between tadpoles of either Bufo americanus Holbrook, 1836 or Rana sylvatica LeConte, 1825 and Echinostoma Rudolphi, 1809 cercariae to understand why echinostomatoid trematodes, such as species from the genera Echinostoma and Ribeiroia Travassos, 1939 (implicated in anuran limb deformities), attack specific anatomical regions of tadpoles. Bufo and Rana tadpoles can shed cercariae on their skin from some parts of their body more easily than others. In particular, cercariae that enter the "dead-water zone" at the junction of a tadpole's body and tail appear particularly difficult for tadpoles to brush off. Cercariae that reach this recess can easily enter the inguinal region of tadpoles (as do Ribeiroia spp.) or ascend the tadpole's cloaca (as do Echinostoma spp.). When tadpoles sense cercariae contacting their skin they make explosive movements to shed those parasites. Factors that reduce tadpoles' activity, such as predator threat or certain pesticides, may increase a tadpole's susceptibility to echinostomatoid infection. Because Bufo tadpoles are unpalatable to many predators, they can afford to make more conspicuous evasive maneuvers than Rana tadpoles, and do so in the laboratory. Bufo tadpoles in the field also have a lower rate and different anatomical distribution pattern of Ribeiroia infection than Rana tadpoles. Factors that reduce tadpole activity in the field may act synergistically to increase parasite loads and subsequent deformities in anurans.

Sire coloration influences offspring survival under predation risk in the moorfrog.

When breeding, male moor frogs Rana arvalis develop a bright blue dorsal coloration which varies in intensity between males. We tested whether this colour acts as a potential signal of a male's genetic quality to female moor frogs by artificially crossing pairs of males differing in the extent of the blue coloration to the same female. Maternal half-sibships provide a powerful means to detect paternal genetic effects on offspring as they control for other potentially confounding variables. We assayed the ability of offspring to survive an ecologically realistic test of fitness by exposing them to predation by the larvae of the predatory water beetle Dytiscus marginalis. Although sire's coloration did not influence tadpole body size, it did affect their ability to survive the predation trial. Offspring of bright blue males had higher survival than those of dull males when exposed to large predators, which were more voracious predators than smaller ones. Our results indicate that paternal secondary sexual traits provide information about genetic effects on offspring fitness in this species, but suggest that these effects may be context-dependent. Variable selection caused by contextual dependence may have important consequences for the evolution of female choice rules, and for the maintenance of genetic variation for both male trait and female preference.