Tooth crown morphology in caecilians (Amphibia: Gymnophiona)

The morphology of tooth crowns is variable inter-specifically among caecilians. Cusp number and shape, crown dimensions, and crown curvature characterize various species and have both functional and phylogenetic implications. Ichthyophis, Uraeotyphlus, Hypogeophis, and Geotrypetes have bicuspid teeth; Dermophis, Gymnopis, Caecilia, and Typhlonectes monocuspid. Crown morphology as revealed by scanning electron microscopy is associated with prey grasping and, in one case, possible specialization of prey type.     

Comparative morphology of caecilian sperm (Amphibia: Gymnophiona)

The morphology of mature sperm from the testes of 22 genera and 29 species representing all five families of caecilians (Amphibia: Gymnophiona) was examined at the light microscope level in order to: (1) determine the effectiveness of silver-staining techniques on long-preserved, rare material, (2) assess the comparative morphology of sperm quantitatively, (3) compare patterns of caecilian sperm morphology with that of other amphibians, and (4) determine if sperm morphology presents any characters useful for systematic analysis. Although patterns of sperm morphology are quite consistent intragenerically and intrafamilially, there are inconsistencies as well. Two major types of sperm occur among caecilians: those with very long heads and pointed acrosomes, and those with shorter, wider heads and blunt acrosomes. Several taxa have sperm with undulating membranes on the flagella, but limitations of the technique likely prevented full determination of tail morphology among all taxa. Cluster analysis is more appropriate for these data than is phylogenetic analysis. © 1994 Wiley-Liss, Inc.     

Evolution of Cranial Shape in Caecilians (Amphibia: Gymnophiona)

Insights into morphological diversification can be obtained from the ways the species of a clade occupy morphospace. Projecting a phylogeny into morphospace provides estimates of evolutionary trajectories as lineages diversified information that can be used to infer the dynamics of evolutionary processes that produced patterns of morphospace occupation. We present here a large-scale investigation into evolution of morphological variation in the skull of caecilian amphibians, a major clade of vertebrates. Because caecilians are limbless, predominantly fossorial animals, diversification of their skull has occurred within a framework imposed by the functional demands of head-first burrowing. We examined cranial shape in 141 species, over half of known species, using X-ray computed tomography and geometric morphometrics. Mapping an existing phylogeny into the cranial morphospace to estimate the history of morphological change (phylomorphospace), we find a striking pattern: most species occupy distinct clusters in cranial morphospace that closely correspond to the main caecilian clades, and each cluster is separated by unoccupied morphospace. The empty spaces in shape space are unlikely to be caused entirely by extinction or incomplete sampling. The main caecilian clades have different amounts of morphological disparity, but neither clade age nor number of species account for this variation. Cranial shape variation is clearly linked to phyletic divergence, but there is also homoplasy, which is attributed to extrinsic factors associated with head-first digging: features of caecilian crania that have been previously argued to correlate with differential microhabitat use and burrowing ability, such as subterminal and terminal mouths, degree of temporal fenestration (stegokrotaphy/zygokrotaphy), and eyes covered by bone, have evolved and many combinations occur in modern species. We find evidence of morphological convergence in cranial shape, among species that have eyes covered by bone, resulting in a narrow bullet-shaped head. These results reveal a complex history, including early expansion of morphospace and both divergent and convergent evolution resulting in the diversity we observe today.     

The heart and aortic arches of rhinatrematid caecilians (Amphibia: Gymnophiona)

Previous studies have shown there to be considerable inter-specific variation in the cardiovascular anatomy of five of the six families of caecilians. Observations on the previously unstudied Rhinatrematidae reveal this family to be characterized by a number of cardiovascular features that are unique within the Gymnophiona. These include a poorly developed sinus venosus sinistra, a short truncus arteriosus, separate carotid and systemic arches and the right atrium larger than the left. Character analysis indicates that these unique features are primitive within the Gymnophiona and they provide considerable additional support for the hypothesis that the Rhinatrematidae are the sister-group to all other caecilians. This hypothesis appears to be among the best supported hypotheses of relationships within the Gymnophiona. Caecilian cardiovascular variation provides a useful source of evidence for phylogeny reconstruction that should be integrated into phylogenetic studies of the group.     

Olfactory and vomeronasal systems of caecilians (Amphibia: Gymnophiona)

The morphology of both the main nasal cavity and the vomeronasal organ differs among species representing six families of caecilians. The main nasal cavity is either divided or undivided. The vomeronasal organ differs in position (mediolateral, lateral), size (large vomeronasal organ in the aquatic species), and shape (mediolateral extension, vomeronasal organ with a lateral rostral projection). The great amount of respiratory epithelium of the main nasal cavity, the large vomeronasal organ, and its extensive innervation in typhlonectids may reflect both phylogeny and habitat adaptation, for these taxa are secondarily aquatic or semiaquatic and have several concomitant morphological and physiological modifications. The vomeronasal organ is associated with the caecilian tentacle as the tentacular ducts open into it. This association is further evidence for the involvement of the caecilian tentacle in vomeronasal chemoperception and may represent the mechanism by which these animals smell though the main nasal cavity is closed during burrowing or swimming. Labelings of primary olfactory and vomeronasal projections by means of horseradish peroxidase reaction reveal that the pattern of vomeronasal projections is similar in Ichthyophis kohtaoensis, Dermophis mexicanus, and Typhlonectes natans, even though T. natans possess stronger vomeronasal projections relative to olfactory projections than I. kohtaoensis and D. mexicanus. However, there are differences with respect to the patterns of olfactory projections. The olfactory projection of I. kohtaoensis is characterized by many displaced glomeruli. T. natans has the smallest olfactory projection. The nervus terminalis is associated with the olfactory system as shown by selective labelings of olfactory projections. Six characters potentially useful for phylogenetic analysis emerge from this study of comparative morphology. The characters were subjected to analysis using PAUP to see (1) if any resolution occurred and (2) if any groups were distinguished, whether they corresponded to phylogenetic arrangements based on other morphological characters. The characters are too few to produce nested dichotomous sets for all cases, but they do support the two typhlonectid genera examined and Dermophis and Gymnopis as sister taxa discrete from other groups, and they show that species within genera cluster together.     

Variation in the trunk musculature of caecilians (Amphibia: Gymnophiona)

Variation in the trunk musculature of 28 species of caecilians. representing 24 of the 33 genera and all five families. is summarized. All forms examined have the same muscles in similar positions. Existing variation largely conforms to the current classification of the group. and some variation may be attributable to different modes of locomotion. such as burrowing versus swimming. Caecilian trunk musculature is more similar to that of salamanders than to that of frogs. but the similarity is probably syrnplesiomorphous. Trunk musculature so far has provided no clues to lissamphibian relationships.     

Cartilage in the cloaca: Phallodeal spicules in Caecilians (Amphibia: Gymnophiona)

The presence of spicules (also termed spines, teeth, or denticles) on the intromittent organ, a character unique to male members of some species in the African caecilian family Scolecomorphidae, has long been known (Noble, '31; Taylor, '68; Wake,'72; Nussbaum,'85). However, their organization and structure has not been examined. Series of males of three species of Scolecomorphus were examined in order to determine the gross and histological organization of the spicules. Spicule morphology changes with ontogeny within species and differs grossly among species. The cellular organization of the spicules is remarkably similar among species, however. The spicules are composed of chondroid cartilage. The large discoid cells are arranged in “stacks” to form the projections, which emerge from an extensive base plate of the same cartilage. The spicules have a connective tissue sheath, which is not continuous with the cloacal epithelium. The sheath of the spicules may or may not be mineralized; the cartilage itself does not appear to be calcified. Mineralization apparently is not correlated with age, reproductive status, or season. Cartilage in the cloaca of these few species poses a series of questions about the interactions of developmental, structural, functional, and phylogenetic properties in the evolution of the reproductive biology of caecilians. J. Morphol. 237:177–186, 1998. © 1998 Wiley-Liss, Inc.     

Marginal plates in hepatic peroxisomes of Ichthyophis glutinosus (Amphibia: Gymnophiona)

Summary The ultrastructure of hepatic peroxisomes was investigated in Ichthyophis glutinosus (Amphibia: Gymnophiona), employing perfusion fixation and the diaminobenzidine (DAB) technique for the visualization of catalase. The majority of peroxisomes is circular or rod-shaped, although elongated particles occasionally occur. They contain a finely granular matrix, lightly stained after the DAB procedure. Their mean diameter is approximately 0.25 m. Serial sections reveal that the circular and rod-shaped peroxisomal profiles are cross and oblique sections of highly tortuous, tubular organelles exceeding 2 m in length.In addition to tubular profiles, elongated, rectangular particles, as well as straight dumbbell-shaped organelles with distinct marginal plates are observed. They range from 900 to 1650 nm in length (mean = 1200 nm). In the flattened, thin central portion of the dumbbell-shaped particle, the peroxisomal membranes form a cisterna enclosing one or two uniformly thick marginal plates, which display a definite substructure with a periodicity of 10 nm.These findings indicate that peroxisomes in the liver of Ichthyophis exhibit a complex organization. It is suggested that the organelles undergo a specific differentiation process, morphologically characterized by the formation of enlarged segments of unusual shape.This study was supported by grants from the Deutsche Forschungsgemeinschaft, Fa 146/1-2 and Sto 75/9     

Copulation and egg retention in an oviparous Caecilian (Amphibia: Gymnophiona)

Viviparity (i.e., the bearing of live young) has evolved from oviparity (egg laying) independently in various major vertebrate lineages, and several transitional stages have been described. The transition from oviparity to viviparity requires the retention of fertilised eggs in the female reproductive tract. Caecilian amphibians (Gymnophiona) display a considerable diversity of reproductive modes, including oviparity and viviparity. Among amphibians, caecilians have also modified the process of internal fertilisation through a special intromittent organ, or phallus, in males. Here we report the oviposition of “embryonated” eggs ranging from various gastrula-to-neurula stages by female Ichthyophis cf. kohtaoensis (Ichthyophiidae) from North-eastern Thailand. In addition, we describe a copulation resulting in an oviposition of embryonated eggs. Our findings will have implications for the further understanding of the evolutionary reproductive biology of amphibians.     

Ultrastructure of the mature spermatozoa of caecilians (Amphibia: Gymnophiona)

The spermatozoa of Gymnophiona show the following autapomorphies: 1) penetration of the distal centriole by the axial fiber; 2) presence of an acrosomal baseplate; 3) presence of an acrosome seat (flattened apical end of nucleus); and 4) absence of juxta-axonemal fibers. The wide separation of the plasma membrane bounding the undulating membrane is here also considered to be apomorphic. Three plesiomorphic spermatozoal characters are recognized that are not seen in other Amphibia but occur in basal amniotes: 1) presence of mitochondria with a delicate array of concentric cristae (concentric cristae of salamander spermatozoa differ in lacking the delicate array); 2) presence of peripheral dense fibers associated with the triplets of the distal centriole; and 3) presence of a simple annulus (a highly modified, elongate annulus is present in salamander sperm). The presence of an endonuclear canal containing a perforatorium is a plesiomorphic feature of caecilian spermatozoa that is shared with urodeles, some basal anurans, sarcopterygian fish, and some amniotes. Spermatozoal synapomorphies are identified for 1) the Uraeotyphlidae and Ichthyophiidae, and 2) the Caeciliidae and Typhlonectidae, suggesting that the members of each pair of families are more closely related to each other than to other caecilians. Although caecilian spermatozoa exhibit the clear amphibian synapomorphy of the unilateral location of the undulating membrane and its axial fiber, they have no apomorphic characters that suggest a closer relationship to either the Urodela or Anura.     

The morphology of prehatching embryos of Caecilia orientalis (Amphibia: Gymnophiona: Caeciliidae)

The state of development of advanced embryos of the direct‐developing Ecuadorian caecilian Caecilia orientalis (Caeciliidae: Gymnophiona: Amphibia) was examined. Because it is established that development is correlated with reproductive modes in a number of features, we included comparison with taxa that represent the major reproductive modes and all of the modern normal tables and ossification sequences. The embryos of C. orientalis most closely resemble those of stage 47/48 Gegeneophis ramaswamii, an Indian caeciliid, and stage 47/48 Hypogeophis rostratus, a Seychellian caeciliid, both direct developers, in details of bone mineralization, chondrocranial degeneration, and vertebrogenesis. They are most like stage 45 H. rostratus in external features (gills, pigmentation, etc.). They are less similar to prehatchings of Ichthyophis kohtaoensis, an ichthyophiid with free‐living larvae, and to fetuses of the viviparous caeciliid Dermophis mexicanus and the viviparous typhlonectid Typhlonectes compressicauda at comparable total lengths in both skeletal development and external features. The similarity of developmental features among the direct‐developers suggests a correlation with mode of life history. A noteworthy feature is that C. orientalis has an armature of multiple rows of teeth on the lower jaw with tooth crowns that resemble the “fetal” teeth of viviparous taxa and that are covered with a layer of oral mucosal epithelium until full development and eruption, but the upper jaw bears a single row of widely spaced, elongate, slightly recurved teeth that resemble those of the adult. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.     

The fine structure of the lateral-line organs of larval Ichthyophis (Amphibia: Gymnophiona)

Light and electron microscopic observations of the lateral-line organs of larval Ichthyophis kohtaoensis confirmed earlier reports of the occurrence of two different types of lateral-line organs. One type, the ampullary organ, possesses 15–26 egg-shaped sensory cells. Each sensory cell extends a single kinocilium surrounded by a few microvilli into the ampullary lumen. This is in contrast to the ampullary organs of urodele amphibians that contain only microvilli. The second type of organ, the ordinary neuromast, has 15–24 pear-shaped sensory cells arranged in two to three rows. Each sensory cell shows a kinocilium that is asymmetrically placed with respect to both a basal plate and approximately 60 stereovilli. The sensory cells of ampullary organs are always separated by supporting cells; those of neuromasts are occasionally in contact with one another. Numerous (neuromasts) or few (ampullary organs) mantle cells separate the organs from the epidermal cells. Only afferent synapses are found in the ampullary organs whereas vesicle-filled fibers together with afferent nerve terminals are found in neuromasts. Both organs contain similarly sized presynaptic spheres adjacent to the afferent fibers. It is suggested that the neuromasts have a mechanoreceptive function, whereas the ampullary organs have an electroreceptive one.     

Mitochondrial evidence on the phylogenetic position of caecilians (Amphibia: Gymnophiona)

The complete nucleotide sequence (17,005 bp) of the mitochondrial genome of the caecilian Typhlonectes natans (Gymnophiona, Amphibia) was determined. This molecule is characterized by two distinctive genomic features: there are seven large 109-bp tandem repeats in the control region, and the sequence for the putative origin of replication of the L strand can potentially fold into two alternative secondary structures (one including part of the tRNA(Cys)). The new sequence data were used to assess the phylogenetic position of caecilians and to gain insights into the origin of living amphibians (frogs, salamanders, and caecilians). Phylogenetic analyses of two data sets-one combining protein-coding genes and the other combining tRNA genes-strongly supported a caecilian + frog clade and, hence, monophyly of modern amphibians. These two data sets could not further resolve relationships among the coelacanth, lungfishes, and tetrapods, but strongly supported diapsid affinities of turtles. Phylogenetic relationships among a larger set of species of frogs, salamanders, and caecilians were estimated with a mitochondrial rRNA data set. Maximum parsimony analysis of this latter data set also recovered monophyly of living amphibians and favored a frog + salamander (Batrachia) relationship. However, bootstrap support was only moderate at these nodes. This is likely due to an extensive among-site rate heterogeneity in the rRNA data set and the narrow window of time in which the three main groups of living amphibians were originated.     

Retinal projections in the caecilian Ichthyophis kohtaoensis (Amphibia,Gymnophiona)

Summary The retinal projections of the caecilian Ichthyophis kohtaoensis were investigated by anterograde transport of HRP. The optic tract forms two bundles in the diencephalon, a narrow medial bundle in the optic tectum, and a basal optic tract consisting of few fibres. Terminal fields are in the thalamus, pretectum, tectum, and as a circum-scribed basal optic neuropile in the tegmentum. Thalamic, pretectal and tectal projections are contralateral as well as ipsilateral. The reduced but existing visual projection corresponds to a reduced but existing visually guided behaviour.     

Morphometric analysis of the skull of Dermophis mexicanus (Amphibia: Gymnophiona)

The levels of integration, patterns of allometric growth and size-related differences in skull dimensions of the caecilian Dermophis mexicanus were investigated by means of univariate, bivariate and multivariate statistics. The analyses indicate that (1) most but not all measurements of the skull arc more variable in adults than in juveniles; (2) growth is allometric and results in changes in shape between juveniles and adults; (3) there is a high level of integration of all variables by general size; and (4) homologous paired measurements show concordant allometric trends and are integrated beyond the sole effect of general size. These analyses provide baseline data, a set of hypotheses and a methodological framework for comparative studies of patterns of variation and integration among caecilians and among other vertebrates.