Unearthing the Fossorial Tadpoles of the Indian Dancing Frog Family Micrixalidae

Tadpoles of the monotypic Indian dancing frog family Micrixalidae have remained obscure for over 125 years. Here we report the discovery of the elusive tadpoles of Micrixalus herrei from the sand beds of a forested stream in southern Western Ghats, and confirm their identity through DNA barcoding. These actively burrowing tadpoles lead an entirely fossorial life from eggs to late metamorphic stages. We describe their internal and external morphological characters while highlighting the following features: eel-like appearance, extensively muscularized body and tail, reduced tail fins, skin-covered eyes, delayed development of eye pigmentation in early pre-metamorphic stages (Gosner stages 25–29), prominent tubular sinistral spiracle, large transverse processes on vertebrae II and III, ankylosed ribs on transverse processes of vertebra II, notochord terminating before the atlantal cotyle-occipital condyle junction, absence of keratodonts, serrated well-formed jaw sheaths, and extensive calcified endolymphatic sacs reaching sacrum posteriorly. The tadpole gut contains mostly fine sediments and sand. We discuss the eel-like morphology and feeding habits of M. herrei in the context of convergence with other well-known fossorial tadpoles. This discovery builds the knowledge base for further comparative analyses and conservation of Micrixalus, an ancient and endemic lineage of Indian frogs.     

The tadpole of Hypsiboas leptolineatus (Braun and Braun, 1977), a species in the Hypsiboas polytaenius clade (Anura; Hylidae).

The larval morphology of Hypsiboas leptolineatus was studied. The tadpole has an ovoid body in lateral view, wider than deep; snout rounded with dorsal reniform nostrils; spiracle sinistral with lateral wall attached to body; anal tube dextral; tail fins convex with acuminate tip; oral disc ventral; labial tooth row formula is 2(2)/3(1); moderately developed beaks with serrated jaw sheaths. These external oral features are compared with those of the known tadpoles in the Hypsiboas polytaenius clade. The oral cavity was studied using an electron microscope. Life history aspects are commented.     

Morphology of the cranial skeleton and musculature in the obligate carnivorous tadpole of Lepidobatrachus laevis (Anura: Ceratophryidae)

Ziermann, J.M., Infante, C., Hanken, J. and Olsson, L. 2011. Morphology of the cranial skeleton and musculature in the obligate carnivorous tadpole of Lepidobatrachus laevis (Anura: Ceratophryidae). —Acta Zoologica (Stockholm) 00 :1–12. Lepidobatrachus laevis (Ceratophryidae: Ceratophryinae) is a bizarre frog endemic to the Chacoan desert of central South America. Its tadpole is an obligate carnivore that can catch and consume live prey nearly its own size. Morphological adaptations associated with this unique feeding mode, including the larval skull anatomy and associated cranial musculature, have only been partly described. We studied the head of Stages 26–27 larvae using gross dissection, immunohistochemistry, and standard histology. Derived features of this tadpole compared to the microphagous, herbivorous larvae of most other anurans include simplified chondrocranial cartilages and very robust jaw muscles. The mm. suspensorio‐ et quadratoangularis do not take their origin from the processus muscularis of the palatoquadrate, as in most other tadpoles, but instead originate from the corpus of the palatoquadrate caudal to this process. The jaw levators are unusually large. The tadpole of Ceratophrys, another member of the ceratophryine clade, also consumes large animal prey, but its morphology is very different. It probably has evolved independently from a generalized, mainly herbivorous tadpole similar to the larva of Chacophrys, the third ceratophryine genus. Most specialized features of the larval head of Lepidobatrachus laevis are adaptations for ‘megalophagy’—ingestion of whole, very large animal prey.     

The forkhead gene FH1 is involved in evolutionary modification of the ascidian tadpole larva.

The forkhead gene FH1 encodes a HNF-3beta protein required for gastrulation and development of chordate features in the ascidian tadpole larva. Although most ascidian species develop via a tadpole larva, the conventional larva has regressed into an anural (tailless) larva in some species. Molgula oculata (the tailed species) exhibits a tadpole larva with chordate features (a dorsal neural sensory organ or otolith, a notochord, striated muscle cells, and a tail), whereas its sister species Molgula occulta (the tailless species) has evolved an anural larva, which has lost these features. Here we examine the role of FH1 in modifying the larval body plan in the tailless species. We also examine FH1 function in tailless speciesxtailed species hybrids, in which the otolith, notochord, and tail are restored. The FH1 gene is expressed primarily in the presumptive endoderm and notochord cells during gastrulation, neurulation, and larval axis formation in both species and hybrids. In the tailless species, FH1 expression is down-regulated after neurulation in concert with arrested otolith, notochord, and tail development. The FH1 expression pattern characteristic of the tailed species is restored in hybrid embryos prior to the development of chordate larval features. Antisense oligodeoxynucleotides (ODNs) shown previously to disrupt FH1 function were used to compare the developmental roles of this gene in both species and hybrids. As described previously, antisense FH1 ODNs inhibited endoderm invagination during gastrulation, notochord extension, and larval tail formation in the tailed species. Antisense FH1 ODNs also affected gastrulation in the tailless species, although the effects were less severe than in the tailed species, and an anural larva was formed. In hybrid embryos, antisense FH1 ODNs blocked restoration of the otolith, notochord, and tail, reverting the larva back to the anural state. The results suggest that changes in FH1 expression are involved in re-organizing the tadpole larva during the evolution of anural development.     

Evolutionary reorganizations of ontogenesis in ascidians of the genus Molgula

The data on comparative, experimental, and molecular embryology of ascidians (genus Molgula) published during the last 15 years have been reviewed. Some representatives of this genus evolved from development with a tailed larva (tadpole) to direct development associated with the loss of larval structures, such as tail, notochord, sensory organs, and differentiated muscles. The data on evolutionary reorganizations of ontogenesis in ascidians of the genus Molgula have been compared with those in sea urchins, anuran amphibians, and some other organisms.     

Developmental dynamics of myogenesis in the shipworm <Emphasis Type="Italic">Lyrodus pedicellatus</Emphasis> (Mollusca: Bivalvia)

Background

The shipworm Lyrodus pedicellatus is a wood-boring bivalve with an unusual vermiform body. Although its larvae are brooded, they retain the general appearance of a typical bivalve veliger-type larva. Here, we describe myogenesis of L. pedicellatus revealed by filamentous actin labelling and discuss the data in a comparative framework in order to test for homologous structures that might be part of the bivalve (larval) muscular ground pattern.

Results

Five major muscle systems were identified: a velum retractor, foot retractor, larval retractor, a distinct mantle musculature and an adductor system. For a short period of larval life, an additional ventral larval retractor is present. Early in development, a velum muscle ring and an oral velum musculature emerge. In late stages the lateral and dorsal mantle musculature, paired finger-shaped muscles, an accessory adductor and a pedal plexus are formed. Similar to other bivalve larvae, L. pedicellatus exhibits three velum retractor muscles, but in contrast to other species, one of them disappears in early stages of L. pedicellatus. The remaining two velum retractors are considerably remodelled during late larval development and are most likely incorporated into the elaborate mantle musculature of the adult.

Conclusions

To our knowledge, this is the first account of any larval retractor system that might contribute to the adult bodyplan of a (conchiferan) mollusk. A comparative analysis shows that a pedal plexus, adductors, a larval velum ring, velum retractors and a ventral larval retractor are commonly found among bivalve larvae, and thus most likely belong to the ground pattern of the bivalve larval musculature.

     

Larval morphology of the scorpionfly Dicerapanorpa magna (Chou) (Mecoptera: Panorpidae) and its adaptive significance

Larval morphology can provide valuable characters for taxonomic and phylogenetic analyses of insects and reflect the adaptations to various living habits. Compared with the adult stages, larval study has lagged far behind in Mecoptera. Although several genera of Panorpidae have been studied for their larval stages, the larva of Dicerapanorpa Zhong and Hua, 2013 basically remains unclear. Here the larva of Dicerapanorpa magna (Chou) is described and illustrated in detail for the first time using light microscopy and scanning electron microscopy. The larva is eruciform, with eight pairs of abdominal prolegs in addition to three pairs of thoracic legs, as in other Panorpidae. The most remarkable characteristics of the larvae include a pair of erect subdorsal annulated processes each on abdominal segments I–IX (A1–A9) and a single middorsal annulated process on A10, as well as a pair of prominent compound eyes composed of over 40 ommatidia, which distinguish this genus from other genera of Panorpidae. The annulated processes may have adaptive significance for fossorial and soil-living habits.     

Evolutionary reorganizations of ontogenesis in ascidians of the genus Molgula

The data on comparative, experimental, and molecular embryology of ascidians (genus Molgula) published during the last 15 years have been reviewed. Some representatives of this genus evolved from development with a tailed larva (tadpole) to direct development associated with the loss of larval structures, such as tail, notochord, sensory organs, and differentiated muscles. The data on evolutionary reorganizations of ontogenesis in ascidians of the genus Molgula have been compared with those in sea urchins, anuran amphibians, and some other organisms.     

Burrow structure and foraging costs in the fossorial rodent,Thomomys bottae

Summary A model for calculating the energy cost of burrowing by fossorial rodents is presented and used to examine the energetics of foraging by burrowing. The pocket gopher Thomomys bottae (Rodentia: Geomyidae) digs burrows for access to food. Feeding tunnels of Thomomys are broken into segments by laterals to the surface that are used to dispose of excavated soil. Energy cost of burrowing depends on both soil type and on burrow structure, defined by the length of burrow segments, angle of ascent of laterals, depth of feeding tunnels, and burrow diameter. In a desert scrub habitat, Thomomys adjust burrow segment length to minimize cost of burrowing. Observed segment lengths (mean=1.33 m) closely approximate the minimum-cost segment length of 1.22 m. Minimizing energy expended per meter of tunnel constructed maximizes efficiency of foraging by burrowing in the desert scrub. Burrow diameter and cost of burrowing increase with body size, while benefits do not, so foraging by burrowing becomes less enconomical as body size increases. Maximum possible body size of fossorial mammals depends on habitat productivity and energy cost of burrowing in local soils.     

A phylogenetic study of the antenna cleaner in Formicidae,Mutillidae, and Tiphiidae (Insecta,Hymenoptera)

Summary The morphology of the tibio-tarsal antenna cleaner (strigilis) of 30 species of Formicidae, 14 species of Mutillidae and 9 species of Tiphiidae was investigated and is described comparatively. In Formicidae, there is a common type of strigilis. The spur has a posterior-dorsal comb and its anterior side is covered with squamae, which usually form a brush. Posteriorly, the basitarsal notch bears a comb and anteriorly specialized paddle-shaped hairs. These characters may be apomorphic for Formicidae. In several Ponerinae and in Myrmecia there is also a velum on the spur. In two species of ants which are strongly parasitic (Teleutomyrmex schneideri and Anergates atratulus ) there are reduced antenna cleaners. Mutillidae (except Myrmosa) have another common type of strigilis: the spur bears a velum with a smooth rim and a clear apex with two rows of teeth. The notch in the basitarsus is usually deeper than that of ants; there is a comb, but no paddle-shaped hairs. The strigilis of Myrmosa females has no velum but there are two prominent rows of teeth on the spur. In the male, the velum is reduced to a slender strip. In Tiphiidae the antenna cleaners show considerable diversity. In Methocha the spur bears a comb but no velum; the spur of Tiphia has a velum with a serrated rim; the spur of Myzinum is equipped with a velum with a smooth rim; in Thynnus the surface of the velum is wrinkled or undulating. An apex (without teeth) is present in all investigated Tiphiidae. The notch of the basitarsus bears a comb except in female Myzinum, where the teeth seem to have fused, thus forming a rim. It is suggested that a velum with a serrated or toothed edge and an apex with two rows of teeth are plesiomorphic for aculeate Hymenoptera. The antenna cleaners in Mutillidae are remarkably similar to those in some bees. This fact is interpreted as partly due to convergence and as partly symplesiomorphic. In several species of ants, there is a specialized cuticular area (bande poreuse) anterior to the comb of the notch, which is characterized by fissures or holes. These are presumably openings of an excretory gland.     

Structure and occurrence of cyphonautes larvae (bryozoa,ectoprocta)

We have studied larvae of the freshwater ctenostome Hislopia malayensis with scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and LM of serial sections. Some additional observations on larvae of M. membranacea using SEM and CLSM are also reported. The overall configuration of muscles, nerves, and cilia of the two larvae are identical. However, the larva of H. malayensis is much smaller than that of M. membranacea, which may explain most of the differences observed. Although all major nerves and muscle strands are present in H. malayensis, they are generally composed of fewer fibers. The H. malayensis larva lacks the anterior and posterior intervalve cilia. Its pyriform organ is unciliated with only a small central depression. The adhesive epithelium is not invaginated as an adhesive sac and lacks the large muscles interpreted as adhesive sac muscles in the M. membranacea larva. The velum carries two rows of ciliated cells, though the lower “row” consists of only one or two cells. Both rows of ciliated cells are innervated by nerves, which have not been detected in the M. membranacea larva. The ciliated ridge of H. malayensis lacks the frontal cilia. The planktotrophic cyphonautes larvae in a number of ctenostome clades and in the “basal” cheilostome clade Malacostega (and probably in the earliest cheilostomes) support the idea that the cyphonautes larva is the ancestral larval type of the Eurystomata. It may even represent the ancestral larval type of the bryozoans (= ectoprocts). J. Morphol. 271:1094‐1109, 2010. © 2010 Wiley‐Liss, Inc.     

The peculiar nemertean larva pilidium recurvatum belongs to Riserius sp., a basal heteronemertean that eats Carcinonemertes errans,a hoplonemertean parasite of Dungeness crab

A typical nemertean pilidium larva resembles a hat with ear flaps. But one type, called pilidium recurvatum, looks more like a sock, swimming heel first. This distinctive larva was discovered in 1883 off the coast of Rhode Island and subsequently found in plankton samples from other parts of the world. Despite the long time since discovery, and its significance in discussions of larval evolution, this larva remained unidentified even to the family level. We collected pilidium recurvatum larvae from plankton samples in Coos Bay, OR, and identified them as belonging to the heteronemertean genus Riserius based on juvenile morphology and DNA sequence data. Phylogenetic analysis suggests that two distinct types of pilidium recurvatum from Oregon represent two new species within this currently monotypic genus. We describe the morphology of pilidium recurvatum using confocal microscopy and compare it to that of the typical pilidium, discussing possible implications for larval feeding. We also report our surprising discovery that juveniles of Riserius sp. from Oregon prey on another nemertean, Carcinonemertes errans, an egg predator of Cancer magister (Dungeness crab), a commercially important species. We speculate that the species‐level diversity and geographic distribution of Riserius may be much greater than currently appreciated.     

Unusual behaviour and morphology of some Rhyacophila Pictet, 1834 caddisfly (Trichoptera: Rhyacophilidae) larvae reflect their ability to use the hyporheic zone

We found that larvae of four Rhyacophila Pictet, 1834 species preferred the hyporheic biotope in rapids and glides, which appears to be an unusual habitat among species in this genus. These species, i.e., Rhyacophila nigrocephala Iwata, 1927, R. nipponica Navas, 1933, R. shikotsuensis Iwata, 1927 and R. kawamurae Tsuda, 1940, belonging to the nigrocephala species group, of which six species occur in Japan. To study the movement behaviour of these species in the hyporheic biotope, we introduced their larvae into a small aquarium with a sandy bottom substrate. The larvae of these species burrowed into the sand bed and moved smoothly through interstices using their forelegs and their highly flexible and elastic abdomens. The larval morphology of these six species differs from that of the representatives of the other species group of Japanese Rhyacophila. They have more slender and flatter head capsules, more elongate abdominal segments, shorter thoracic legs and more slender anal prolegs. These features are adaptations that allow these species to effectively use the hyporheic biotope by enabling them to burrow through the interstices.     

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

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

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.