A scanning electron microscope study of the odontoids and teeth in Hemiphractus proboscideus (Anura: Hylidae)

In Hemiphractus proboscideus odontoids are found on the dentary, angular and palatine bones. The morphology of all odontoids is similar, although there are minor variations. They grow by appositional growth, probably quite slowly and possibly seasonally. The odontoids are produced as a result of increase in width of the lamellae near the surface of their supporting bone, and seem to be capable of repair after damage in vivo . The teeth on the premaxilla, maxilla and prevomer are strongly recurved and are composed of dentine covered over their apical one-fifth by a thin layer of aprismatic enamel. The teeth are monocuspid but each possesses two small tubercles which are situated on the mesial and distal margins near the apex. The surface dentine is composed of longitudinal bundles of calcified fibres connected by horizontal interlocking fibres. During tooth replacement resorption bays are visible on the external and pulpal surfaces of the dentine.     

A cladistic analysis of Scinax (Anura: Hylidae)

The hylid frog genus Scinax ranges from southern Mexico to eastern Argentina. The 84 recognized species are currently arranged in five groups: the Scinax rubra , Scinax rostrata , Scinax catharinae , Scinax staufferi , and Scinax perpusilla species groups. Although a few species groups are distinctive, compelling evidence of monophyly is lacking for three groups, and their interrelationships remain virtually unknown. With the objective of testing the monophyly of the species groups and exploring their interrelationships, a cladistic analysis was performed, employing 86 characters from osteology, myology, adult and larval external morphology, reproductive biology, and karyotypes taken from 36 representative species of the five species groups plus eight outgroup taxa. The analysis of the resulting data matrix shows moderately well-supported (as assessed through Bremer support and parsimony jackknifing) S. catharinae and S. rostrata species groups. The S. staufferi species group is polyphyletic, as currently composed, given that some of its species are nested within the S. catharinae species group. The only representative of the S. perpusilla species group is the sister species of the S. catharinae species group. Although weakly supported, the representatives of the S. rostrata and S. staufferi species groups are nested, separately, within the S. rubra species group. In order to have a taxonomy consistent with these results, it is proposed not to recognize the S. rubra group but to recognize a more inclusive rubra clade that includes all studied exemplars of the former S. rubra group, plus some of the S. staufferi group and all of the S. rostrata group. Similarly, a catharinae clade is recognized, within which all studied exemplars of the S. catharina e group (that now includes Scinax berthae and Scinax agilis ) and the one studied representative of the S. perpusilla group are included.     

The phylogeny of Dendropsophini (Anura: Hylidae: Hylinae)

The relationships of the hyline tribe Dendropsophini remain poorly studied, with most published analyses dealing with few of the species groups of Dendropsophus. In order to test the monophyly of Dendropsophini, its genera, and the species groups currently recognized in Dendropsophus, we performed a total evidence phylogenetic analysis. The molecular dataset included sequences of three mitochondrial and five nuclear genes from 210 terminals, including 12 outgroup species, the two species of Xenohyla, and 93 of the 108 recognized species of Dendropsophus. The phenomic dataset includes 46 terminals, one per species (34 Dendropsophus, one Xenohyla, and 11 outgroup species). Our results corroborate the monophyly of Dendropsophini and the reciprocal monophyly of Dendropsophus and Xenohyla. Some species groups of Dendropsophus are paraphyletic (the D. microcephalus, D. minimus, and D. parviceps groups, and the D. rubicundulus clade). On the basis of our results, we recognize nine species groups; for three of them (D. leucophyllatus, D. microcephalus, and D. parviceps groups) we recognize some nominal clades to highlight specific morphology or relationships and facilitate species taxonomy. We further discuss the evolution of oviposition site selection, where our results show multiple instances of independent evolution of terrestrial egg clutches during the evolutionary history of Dendropsophus.     

Meiotic chromosomes of the tree frog Smilisca baudinii (Anura: Hylidae)

The Mexican tree frog Smilisca baudinii, is a very common frog in Central America. In spite their importance to keep the ecological equilibrium of the rainforest, its biology and genetics are poorly known. In order to contribute with its biological knowledge, we described the typical meiotic karyotype based in standard cytogenetic protocols to specimens collected in Tabasco, Mexico. The study was centered in the analysis of 131 chromosome spreads at meiotic stage from two adults of the species (one female and one male). The metaphase analysis allowed the establishment of the modal haploid number of 1n = 12 bivalent chromosomes. The chromosomic formulae from the haploid bivalent karyotype was integrated by 12 biarmed chromosomes characterized by twelve pairs of metacentric-submetacentric (msm) chromosomes. The meiotic counting gives the idea that diploid chromosome number is integrated by a complement of 2n = 24 biarmed chromosomes. The presence of sex chromosomes from female and male meiotic spreads was not observed. Current results suggest that S. baudinii chromosome structure is well shared among Hylidae family and "B" chromosomes are particular structures that have very important evolutionary consequences in species diversification.     

The tadpole of Scinax aromothyella (Anura: Hylidae) from Uruguay

We describe the tadpole of Scinax aromothyella from the surroundings of Quebrada de los Cuervos, Departamento de Treinta y Tres, Uruguay. It shares with other larvae of the Scinax catharinae clade the slightly depressed body, a dextral vent tube that reaches the free margin of the ventral fin, and the oral disc with a dorsal gap in the marginal row of papillae and with a concave posterior edge. It is most similar to the tadpole of Scinax berthae, from which it differs in the larger total length of S. aromothyella.

Describimos la larva de Scinax aromothyella de los alrededores de la Quebrada de los Cuervos, Departamento de Treinta y Tres, Uruguay. Comparte con las larvas de otras especies del clado de Scinax catharinae el tener el cuerpo levemente deprimido, el tubo proctodeal dextro que alcanza el borde libre de la aleta ventral, y el disco oral con un claro rostral en la hilera de papilas marginales y con su borde posterior cóncavo. Es muy parecida a la larva de Scinax berthae, de la que difiere en la mayor longitud total de S. aromothyella.     

The submandibular musculature of phyllomedusinae (Anura: Hylidae): A reappraisal

The submandibular musculature of 37 species of the five currently recognized genera of the subfamily Phyllomedusinae (Anura: Hylidae) is described; observations are made on the variation and ontogeny of these muscles. Supplementary apical elements of the m. intermandibularis occur in all phyllomedusines studied, in addition to the supplementary posterolateral elements previously reported. Our observations are discussed in the context of 1) the proposed homology between supplementary apical and posterolateral elements; 2) the homology with the apical elements reported for Pelodryadinae (sister taxon of Phyllomedusinae); and 3) the implications for our understanding of the relationships between Phyllomedusinae and Pelodryadinae. Anatomical differences between the apical and posterolateral elements and their co‐occurrence in phyllomedusines indicate that these supplementary elements are not homologous. Despite differences between phyllomedusines and pelodryadines in the adhesion of supplementary fibers to the principal element of the m. intermandibularis and the occurrence of a broad aponeurosis or a medial raphe, the extensive morphological and developmental resemblances of the apical elements indicate that these structures are homologous, and that the presence of apical elements is a synapomorphy of Phyllomedusinae + Pelodryadinae. J. Morphol., 2011. © 2011 Wiley‐Liss, Inc.     

Muscle development in the abdominal region of Larval Hylidae (Amphibia: Anura)

Larval muscle development in the abdominal region of five species of hylid frogs (Scinax nasicum, S. fuscovarium, Hyla andina, Phyllomedusa boliviana, Gastrotheca gracilis) was studied using differential staining techniques. These five species represent three major hylid subfamilies. The development of the main abdominal muscles, the rectus abdominis, the two lateral muscles (obliquus externus and transversus), and the lateral pectoralis abdominalis is described. The number of myotomes of the rectus abdominis varies between five and six, and the abdominal muscles associated with the rectus abdominis (obliquus externus, pectoralis abdominalis, and rectus cervicis) vary interspecifically in time of appearance and configuration. The presence of gaps in the configuration of the rectus abdominis has been related to the lotic habits of the larvae. However, our observations indicate the presence of such gaps in larvae that inhabit lentic environments as well. These results suggest that the presence of these gaps is unrelated to larval habitat. There are relatively small differences in muscle morphology among these closely related species, which apparently cannot be explained by morphological adaptations related to their ecology. In the species studied, the number of elements that form the abdominal musculature in larvae is equal to that observed in adults. Likewise, the general morphology of the muscles is ontogenetically conserved. This suggests that both the axial skeleton and musculature are more ontogenetically conserved in relation to the substantial changes that are observed in the skull and head muscles of developing anurans. J. Morphol. 241:275–282, 1999. © 1999 Wiley‐Liss, Inc.     

Structural diversity of nuptial pads in Phyllomedusinae (Amphibia: Anura: Hylidae)

We studied the morphological variation of the nuptial pads using light microscopy and scanning electron microscopy (SEM) in 26 species of phyllomedusines (Anura: Hylidae), representing the five currently recognized genera. All phyllomedusines have single nuptial pads with dark colored epidermal projections (EPs). Spine‐shaped EPs occur in Cruziohyla calcarifer, Phrynomedusa appendiculata and in one species of Phasmahyla. The other species have roundish EPs. The density of the EPs on the pad is variable. Species in the Phyllomedusa hypochondrialis Group have EPs with a density that varies between 764 ± 58/mm² and 923 ± 160/mm². In all other studied species (including the Phyllomedusa burmeisteri and Phyllomedusa perinesos groups, Phyllomedusa camba, Phyllomedusa boliviana, Phyllomedusa sauvagii, Phyllomedusa bicolor, and Phyllomedusa tomopterna) the density of EPs varies between 108 ± 20/mm² and 552 ± 97/mm². Pores were observed with SEM in C. calcarifer, Agalychnis lemur, Agalychnis moreletii, but its presence is confirmed through histological sections on several other species. Its visibility using SEM seems to be related with the level of separation between adjacent EPs. The pores in the four studied species of Agalychnis are shown with SEM and histological sections to have a characteristic epidermal rim, that is absent in the otherphyllomedusines. Unlike most previous reports on breeding glands, those of phyllomedusines are alcian blue positive, indicating the presence of acidic mucosubstances on its secretions. J. Morphol. 2012. © 2012 Wiley Periodicals, Inc.     

A new species of Osteocephalus (Anura: Hylidae) from Guyana.

A new member of the genus Osteocephalus is described from the Pakaraima mountains of western Guyana. This species is the smallest known member of the genus and is probably closely related to O. subtilis. Both share a small size (less than 40 mm snout-vent length), large and bulgy eyes directed somewhat rostrally, green bones, smooth and brownish dorsal skin, relatively short and truncate snout, small tympanum, subgular and laterally expanded vocal sac, poorly developed subarticular and supernumerary tubercles, a supra-anal glandular ridge, and cream-white venter and subocular region. The new species can be distinguished from O. subtilis by the Buff iris (vs black), smaller overall size (32.7 vs 35.8-38.8 mm snout-vent length), relatively larger toe disks, and less developed foot webbing. The cranium of the new species is well ossified, relatively reduced in width between the orbits, without an exposed frontoparietal fontanelle and with the anterior arm of the squamosal extending to about half the distance to the maxillary. The vocal sac is subgularly poorly developed and possess lateral extensions to the area behind the jaw angles. Well developed supraocular and suprasquamosal cartilages give support to the enlarged eyes of this species.     

The hyolaryngeal apparatus of three species of Scinax (Scinaxinae: Hylidae: Anura) and comparison with other Hylidae species

The hyolaryngeal apparatus includes the arytenoid, cricoid and hyoid cartilages, and varied sizes and shapes can be distinguished between species. Although the larynx is an important morphological structure for sound production in frogs, few studies describe the structures and processes of its parts in hylids. We studied the hyolaryngeal anatomy of both males and females of Scinax ruber (Laurenti, 1768), using clearing and double staining methods, comparing it with that of Scinax wandae (Pyburn and Fouquette, 1971), Scinax kennedyi (Pyburn, 1973) and other hylid species. We found that S. ruber has the largest arytenoid cartilage of any species of the subfamily Scinaxinae studied to date. We described both the laryngeal differences among three Scinax species and the interspecific variability in the shapes of the projections of the arytenoid in the males of these species. The taxonomic characters (presence/absence of processes of the cartilage of Santorini, oesophageal process or bronchial processes) described here can play a key role in differentiating these species from Scinax species and from other species. In addition, we want to contribute to the natural history and comparison of the larynges in Scinax and within the genus and the Hylidae.     

Vocalizations of Hypsiboas goianus (Lutz, 1968) (Anura: Hylidae) in Central Brazil

Environmental and morphological factors and social context influence the vocalizations of anuran species. Herein, we observed the acoustic behavior of Hypsiboas goianus males from Central Brazil and analyzed the following calls: advertisement, short aggressive and long aggressive. The acoustic parameters of advertisement calls were not correlated with air temperature, mass or snout–vent length; however, the acoustic parameters of short aggressive calls were influenced by these variables. The temporal parameters of advertisement and short aggressive calls were classified as dynamic properties, while the dominant frequency was considered a static property. Dominant frequency is the most important variable to discriminate calls among individuals.     

Phylogenetic relationships and biodiversity in Hylids (Anura: Hylidae) from French Guiana

We evaluated two biodiversity criteria, higher taxonomic diversity and phylogenetic diversity in French Guiana. For this, we used a recent assessment of the knowledge accumulated since 30 years of study on the amphibian species currently known in French Guiana. We focused on two well-represented genera, Hyla and Scinax, belonging to the subfamily Hylinae. We used partial sequences of two mitochondrial genes (16S rDNA and 12S rDNA, 813 bp) and two nuclear genes (tyrosinase and 18S rRNA, 1590 bp) covering a total of 2403 bp. According to the high bootstrap support in phylogenetic analysis of the complete dataset, the genus Scinax is a homophyletic clade formed by two species groups (rubra and rostrata) in French Guiana. The genus Hyla was confirmed to be a paraphyletic group formed by two species groups as well (30 chromosomes and the 'gladiator frogs'). We confirmed that these genera should be taxonomically reconsidered. Moreover, at the genus, subfamily and family levels, the use of only morphological characters or only molecular DNA markers would hamper estimations of biodiversity. Thus, we strongly advise the combined use of both morphology and molecular data (nuclear and mitochondrial markers).     

Conserved karyotypes in the Hyla pulchella species group (Anura, Hylidae)

Cytogenetic analyses were done on specimens of Hyla marginata and on three populations of H. semiguttata differing in morphology and in the physical parameters of their advertisement call, as well as in individuals of Hyla sp. (aff. semiguttata). All specimens had 2n=24 chromosomes with a morphology very similar to that of other 24-chromosome Hyla species. Hyla semiguttata and H. marginata showed the same C-banding pattern but were distinguished by the location of the NOR on pair 1 in H. semiguttata (in the three populations) and Hyla sp. (aff. semiguttata), and on pair 10 in H. marginata. The H. semiguttata populations did not differ cytogenetically, despite variations in their morphology and advertisement calls. Similarly, H. semiguttata and H. p. joaquini studied previously had identical C-banding patterns and NOR locations, suggesting that they are very closely related.     

Comparative skeletogenesis of the Oriental Tree Frog Hyla orientalis (Anura: Hylidae)

The ontogeny of skeletal development of the Oriental Tree Frog, Hyla orientalis (Bedriaga, 1890), is followed from tadpole to froglet and compared with four other hylid species i.e., Hypsiboas lanciformis, Hypsiboas pulchellus, Phyllomedusa vaillanti and Pseudis platensis. Our analysis and comparison is based on cleared and double-stained specimens. The parasphenoid is the first skeletal element that ossified in Hyla orientalis is followed by the exoocipital, prootic, axial and appendicular skeletal elements, frontoparietal, nasal, upper jaw, squamosal, pterygoid, lower jaw, vomer and quadratojugal. Ossification initiated at early Stage 33 in Hyla orientalis whereas in other hylids it begins after Stage 37. Major transformations of the larval jaw were visible after Stage 42. Cranial ossifications were completed after appendicular and axial ossifications of Hyla orientalis. All skeletal elements ossified before the end of the metamorphosis, a unique feature of Hyla orientalis in comparison to the other hylids studied here.     

A total evidence analysis of the phylogeny of hatchet-faced treefrogs (Anura: Hylidae: Sphaenorhynchus)

The Neotropical hylid genus Sphaenorhynchus includes 15 species of small, greenish treefrogs widespread in the Amazon and Orinoco basins, and in the Atlantic Forest of Brazil. Although some studies have addressed the phylogenetic relationships of the genus with other hylids using a few exemplar species, its internal relationships remain poorly understood. In order to test its monophyly and the relationships among its species, we performed a total evidence phylogenetic analysis of sequences of three mitochondrial and three nuclear genes, and 193 phenotypic characters from all species of Sphaenorhynchus. Our results support the monophyly of Sphaenorhynchus with molecular and phenotypic evidence, with S. pauloalvini as the earliest diverging taxon, followed by S. carneus, as the sister taxon of all remaining species of the genus. We recognize three species groups in Sphaenorhynchus (the S. lacteus, S. planicola and S. platycephalus groups), to facilitate its taxonomic study; only three species (S. carneus, S. pauloalvini and S. prasinus) remain unassigned to any group. Sequence data were not available for only two species (S. bromelicola and S. palustris) for which we scored phenotypic data; wildcard behaviour was detected only in S. bromelicola nested inside the S. platycephalus group. On the basis of the resulting phylogenetic hypothesis, we discuss the evolution of oviposition site and a number of phenotypic characters that could be associated with heterochronic events in the evolutionary history of this group.     

Germ cell dynamics during the annual reproductive cycle of Dendropsophus minutus (Anura: Hylidae)

Thirty male specimens of Dendropsophus minutus Peters, 1972, were collected from April 2004 to March 2005 in the region de Sao José do Rio Preto/SP, to conduct a histological study during the seasonal and annual cycles. Testicular activity was inferred based on the volume occupied by each type of cellular cyst present in the seminiferous tubules, as well as the quantity of germ cells in the final development stage, the spermatozoids. All data analyzed were correlated with climatic variables (temperature, rainfall and photoperiod) registered in the region where specimens were collected. A significant variation was verified in the quantity of spermatozoids as well as in the volume occupied by spermatids and spermatozoids throughout the year and between the cold/dry and hot/ humid seasons. It has also been reported that environmental conditions are important factors closely related to species reproduction and that production of germ cells and volume occupied by germ cysts is independent of anatomical aspect of the gonads. Thus, it was possible to verify that although the species reproduces throughout the year, individuals exhibit a preferential reproduction season, resulting in a reproductive (October to the end of February) and a post-reproductive period.