A new species of Hatchet-faced Treefrog Sphaenorhynchus Tschudi (Anura: Hylidae) from Quadrilátero Ferrífero, Minas Gerais, southeastern Brazil.

A new species of Sphaenorhynchus is described from the Municipality of Mariana, State of Minas Gerais, Brazil. It is characterized by the vocal sac moderately developed, single, subgular, with longitudinal folds; white canthal and dorsolateral lines delimited below by a dorsolateral black line from the tip of snout extending beyond the eye to gradually disappearing up to the flanks; and premaxilla and maxilla almost completely edentulous, each bearing 1-5 extremely small teeth. It is most similar with Sphaenorhynchus orophilus, from which it can be distinguished by having a less robust forearm in males; glandular subcloacal dermal fold; premaxilla and maxilla almost completely edentulous; and larvae with large marginal papillae in the oral disc. The new species occurs in natural ponds over ironstone outcrops (known as canga) on flat terrain, where males call from the floating vegetation.

A new rainfrog of the Pristimantis myersi Group (Amphibia, Craugastoridae) from Volcán Pichincha, Ecuador.

A new frog of the Pristimantis myersi Group is described from a bamboo patch within the Reserva Ecológica Verdecocha (0°5'46.9"S, 78°36'15.3"W; 2851 m), located at northwestern flank of the Volcán Pichincha, in the vicinities of Quito, Ecuador. The new species is known from eight adult males, whereas the females remain unknown; it can be readily distinguished from all species of the P. myersi Group that inhabit the highlands of the Ecuadorian Andes by the unique combination of the following characters: body small (adult male SVL 14.9-19.7 mm; females unknown); dorsal skin shagreen, with a barely visible middorsal raphe, scapular and dorsolateral folds; tympanum small but well-defined; upper eyelid with one enlarged tubercle; males with prominent vocal slits, but without nuptial pads on thumbs; fold-like tarsal tubercles. With this new species, the number of Pristimantis assigned to the P. myersi Group raises to 16, of which, 12 are in Ecuador. We provide notes on morphology and color variation, advertisement call, and natural history of the new species.

The gastrocoel roof plate in embryos of different frogs.

The morphology of the gastrocoel roof plate and the presence of cilia in this structure were examined in embryos of four species of frogs. Embryos of Ceratophrys stolzmanni (Ceratophryidae) and Engystomops randi (Leiuperidae) develop rapidly, provide comparison for the analysis of gastrocoel roof plate development in the slow-developing embryos of Epipedobates machalilla (Dendrobatidae) and Gastrotheca riobambae (Hemiphractidae). Embryos of the analyzed frogs develop from eggs of different sizes, and display different reproductive and developmental strategies. In particular, dorsal convergence and extension and archenteron elongation begin during gastrulation in embryos of rapidly developing frogs, as in Xenopus laevis. In contrast, cells that involute during gastrulation are stored in the large circumblastoporal collar that develops around the closed blastopore in embryos of slow-developing frogs. Dorsal convergence and extension only start after blastopore closure in slow-developing frog embryos. However, in the neurulae, a gastrocoel roof plate develops, despite the accumulation of superficial mesodermal cells in the circumblastoporal collar. Embryos of all four species develop a ciliated gastrocoel roof plate at the beginning of neurulation. Accordingly, fluid-flow across the gastrocoel roof plate is likely the mechanism of left-right asymmetry patterning in these frogs, as in X. laevis and other vertebrates. A ciliated gastrocoel roof plate, with a likely origin as superficial mesoderm, is conserved in frogs belonging to four different families and with different modes of gastrulation.

Life in the slow lane: the effect of reduced mobility on tadpole limb development.

Movement is thought to be a primary agent eliciting basic responses in the vertebrate body, such as the proper development of the musculoskeletal system. Embryos do not passively await hatching or birth but rather begin active movement very early on in their development. Most studies dealing with embryonic responses to changes in mobility have been performed in chickens or mammals. Herein, we investigate for the first time whether the embryos of organisms that are free-living during development demonstrate the same morphological responses to reduced mobility as embryos that undergo development in controlled environments such as in utero or in a shelled egg. We changed the viscosity of the environment in which free-living anuran tadpoles grow by rearing them in an agar medium. We thus increased the viscosity of the growth medium resulting in a decrease in larval movement. We predicted that a substantial increase in viscosity of the medium in which the larvae were reared would have at least two consequences: (1) a reduction of tadpole mobility and (2) a delayed onset of skeletogenesis thus producing shorter long bones. Our predictions were upheld and tadpoles reared in an agar medium remain immobile longer and showed a delayed onset of skeletogenesis compared with controls. We propose that the developmental responses to the same stimulus are similar throughout tetrapods, regardless of their developmental context (i.e., intrauterine, within an egg, or free-living).

Influence of forest fragmentation on community structure of frogs and lizards in northeastern Costa Rica.

To better understand responses of reptiles and amphibians to forest fragmentation in the lowland Neotropics, we examined community and population structure of frogs and lizards in the fragmented landscape surrounding La Selva Biological Station in the Sarapiquí region of northeastern Costa Rica. We used diurnal quadrats and nocturnal transects to sample frogs and lizards in nine forest fragments (1-7 ha each) and La Selva (1100 ha). Species richness in all fragments combined was 85% of that found in La Selva with comparable sampling effort. Richness varied from 10 to 24 species among forest fragments, compared with 36 species at La Selva. Lizard density was higher and frog density was lower in forest fragments than in La Selva. Community composition varied among sites and by fragment size class, and species occurrence was nested with respect to fragment area. Isolation and habitat variables did not significantly affect species richness, composition, or nestedness. We classified 34% of species as fragmentation sensitive because they were absent or occurred at low densities in fragments. Nevertheless, the relatively high diversity observed in the entire set of fragments indicates that preserving a network of small forest patches may be of considerable conservation value to the amphibians and reptiles of this region.

Digestive plasticity in tadpoles of the Chilean giant frog (Caudiverbera caudiverbera): factorial effects of diet and temperature.

Anuran metamorphosis is one of the most spectacular processes in nature. Metamorphosis entails morphological transformations and extensive changes in feeding habits, such as transforming from an herbivore to a carnivore. This phenomenon is especially sensitive to environmental cues. We studied the phenotypic plasticity of intestinal morphology and enzyme activity in tadpoles of the Chilean giant frog Caudiverbera caudiverbera. We tested the effects of diet and temperature using a factorial design, which included a control of nontreated individuals. There was no significant effect of diet treatment (i.e., low- vs. high-quality diet) on any of the measured variables, including external morphology. We found significant effects of temperature on morphological traits. Temperature treatment also had significant effects on aminopeptidase-N and maltase activity. Both enzymes exhibited complex interactions with temperature along the intestine. Gut size varied significantly among temperatures, with intestines from warm-treated individuals smaller than the intestines from control and cold-treated tadpoles. Our findings suggest that phenotypic plasticity of intestinal morphology and physiology exists in larvae of this species, at least in response to temperature. However, we did not detect clear effects of diet or temperature on the timing of metamorphosis.

Micronucleus induction in erythrocytes of the Hyla pulchella tadpoles (Amphibia: Hylidae) exposed to insecticide endosulfan.

Endosulfan is a synthetic chlorinated and environmental genotoxic pesticide used worldwide for crop production. We used the micronucleus test in erythrocytes of Hyla pulchella tadpoles in order to develop an experimental model for detecting genotoxic effects of the synthetic chlorinated cyclodiene endosulfan. The frequency of micronuclei was examined in blood smears obtained from tadpoles exposed in vivo to three different concentrations 2.5, 5, and 10 microg/l of the compound and fixed at two sampling times 48 and 96 h. As a positive control larvae were exposed to 40 mg/l of cyclophosphamide. Results obtained here demonstrated the genotoxic effects of the commercial formulation endosulfan in the experimental model assayed.

Effects of behavioral and morphological plasticity on risk of predation in a Neotropical tadpole.

Predator-induced phenotypic plasticity is widespread among aquatic animals, however the relative contributions of behavioral and morphological shifts to reducing risk of predation remain uncertain. We tested the phenotypic plasticity of a Neotropical tadpole ( Rana palmipes) in response to chemical cues from predatory Belostoma water bugs, and how phenotype affects risk of predation. Behavior, morphology, and pigmentation all were plastic, resulting in a predator-induced phenotype with lower activity, deeper tail fin and muscle, and darker pigmentation. Tadpoles in the predator cue treatment also grew more rapidly, possibly as a result of the nutrient subsidy from feeding the caged predator. For comparison to phenotypes induced in the experiment, we quantified the phenotype of tadpoles from a natural pool. Wild-caught tadpoles did not match either experimentally induced phenotype; their morphology was more similar to that produced in the control treatment, but their low swimming activity was similar to that induced by predator cues. Exposure of tadpoles from both experimental treatments and the natural pool to a free-ranging predator confirmed that predator-induced phenotypic plasticity reduces risk of predation. Risk of predation was comparable among wild-caught and predator-induced tadpoles, indicating that behavioral shifts can substantially alleviate risk in tadpoles that lack the typical suite of predator-induced morphological traits. The morphology observed in wild-caught tadpoles is associated with rapid growth and high competition in other tadpole species, suggesting that tadpoles may profitably combine a morphology suited to competition for food with behaviors that minimize risk of predation.

Description of the previously unknown tadpole of Hyalinobatrachium pulveratum (Anura: Centrolenidae).

Egg clutches of the centrolenid Hyalinobatrachium pulveratum were sampled in four lowland locations of Costa Rica. The ontogenesis of the tadpoles of this species is documented by periodical descriptions of the larval stages. Larvae of H. pulveratum change their shape during development because of the non-equal growth of some body parts. Due to these changes the larvae in early stages of development differ considerably to those from later stages. Young larvae have a nearly circular cross-section, whereas later larval stages change to a typical flat shape. Some further morphological characteristics like width of the interorbital distance and the oral disk width change but not proportional to larvae length. Numerous measurements on different larval sizes help to recognize the larvae of H. pulveratum in all stages. The striking bright red hyobranchial sinus is the conspicuous characteristic feature ventrally of the hyobranchial apparatus. The use of allometric values for taxonomic and ontogenetic studies is discussed.

Functional expression of Kir 6.1/SUR1-K(ATP) channels in frog retinal Müller glial cells.

The retinae and brains of larval and adult amphibians survive long-lasting anoxia; this finding suggests the presence of functional K(ATP) channels. We have previously shown with immunocytochemistry studies that retinal glial (Müller) cells in adult frogs express the K(ATP) channel and receptor proteins, Kir6.1 and SUR1, while retinal neurons display Kir6.2 and SUR2A/B (Skatchkov et al., 2001a: NeuroReport 12:1437-1441; Eaton et al., in press: NeuroReport). Using both immunocytochemistry and electrophysiology, we demonstrate the expression of Kir6.1/SUR1 (K(ATP)) channels in adult frog and tadpole Müller cells. Using conditions favoring the activation of K(ATP) channels (i.e., ATP- and spermine-free cytoplasm-dialyzing solution containing gluconate) in Müller cells isolated from both adult frogs and tadpoles, we demonstrate the following. First, using the patch-clamp technique in whole-cell recordings, tolbutamide, a blocker of K(ATP) channels, blocks nearly 100% of the transient and about 30% of the steady-state inward currents and depolarizes the cell membrane by 5-12 mV. Second, inside-out membrane patches display a single-channel inward current induced by gluconate (40 mM) and blocked by ATP (200 microM) at the cytoplasmic side. The channels apparently show two sublevels (each of approximately 27-32 pS) with a total of 85-pS maximal conductance at -80 mV; the open probability follows a two-exponential mechanism. Thus, functional K(ATP) channels, composed of Kir6.1/SUR1, are present in frog Müller cells and contribute a significant part to the whole-cell K+ inward currents in the absence of ATP. Other inwardly rectifying channels, such as Kir4.1 or Kir2.1, may mediate the remaining currents. K(ATP) channels may help maintain glial cell functions during ATP deficiency.