Intraspecific variation in laryngeal and ear morphology in male cricket frogs (Acris crepitans)

In a previous report, the authors found significant population variation in the calls of cricket frogs ( Acris crepitans ) that could not be explained by geographic variation in body size alone. Here we extend that work by investigating intraspecific population variation in the morphological characteristics underlying acoustic communication in male cricket frogs from several sites in Texas. We measured the volumes of laryngeal and auditory components responsible for the generation or reception of species-specific vocalizations in male frogs from eight populations. We found significant differences among populations in body size, as well as all the laryngeal and ear components we measured. With the exception of vocal cord and extracolumella volumes, the volumes of these anatomical structures differ among populations independently of body size as determined by a covariate analysis with snout-vent length as the covariate. Call dominant frequency differs among populations in a clinal pattern and head width, arytenoid cartilage, vocal cord and dilator muscle volume show a similar pattern when the residuals of the regression of morphological component on SVL are assessed for this trend. The results show that both larynx and ear structures can change in size independently of body size, yielding significant geographic variation in the behavioral and physiological expressions of the acoustic communication system underlying mate choice.     

Evolution of intraspecific variation in the advertisement call of a cricket frog (Acris crepitans, Hylidae)

Advertisement calls of the cricket frog, Acris crepitans , show statistically significant variation among populations in all call variables measured. Call variables show strong clinal variation resulting in calls of lower frequency, longer duration and slower call rates produced by A. c. blanchardi in open habitat in the west of the range, and calls of higher frequency, shorter calls and faster call rates produced by A. c. crepitans in the pine forests in the eastern part of the range. This clinal variation does not result from pleiotropic effects of body size or any other morphological characters we measured.
The two subspecies usually reside in different habitats, but some A. c. blanchardi reside in an isolated pine forest in central Texas. By comparing the calls of this subspecies in open and forest habitat, and by statistically removing the effects of clinal variation for all populations, we determined that habitat explains some of the variation in call structure; this is not true of subspecies.
Our data reject several hypotheses that purport to explain the evolution of mate recognition signals. (1) We reject the notion of Paterson and others that there is strong stabilizing selection on species-specific mate recognition signals. (2) There is no support for the hypothesis that call variation is primarily due to pleiotropic effects of body size or other morphological characters over the geographic range we examined. (3) There is no evidence for reproductive character displacement. (4) Our data, as well as experimental studies of habitat acoustics, support the hypothesis that some differences in calls among habitats result from environmental selection on call structure to enhance call transmission. We suggest that the latter hypothesis does not explain the strong clinal component of call variation. This might result from the passive effects of gene flow between populations at the extremes of the range under selection generated by habitat acoustics.     

Trade-off in short- and long-distance communication in tungara (Physalaemus pustulosus) and cricket (Acris crepitans) frogs

Female phonotaxis in túngara (Physalaemus pustulosus)and cricket (Acris crepitans) frogs is biased toward male advertisementcalls or call components of lower frequency. This behavioralbias might result in part from a mismatch between the spectralcharacteristics of the advertisement call and the most sensitive frequencyof the peripheral end organ implicated in reception of thesesounds. In both species, females are tuned to frequencies lowerthan average for the calls in their population. This mismatch,however, represents the situation during short-distance communication.Female frogs can also use the call to detect choruses at longdistances, and the spectral distribution of call energy canvary with transmission distance. We used computer simulationsto test the hypothesis that there is a better match betweentuning and call spectral energy at long distances from the callingmale than at short distances by comparing the performance (soundenergy received) of the natural tuning curve relative to anoptimal tuning curve (i.e., one centered at the call's dominantfrequency). The relative performance of the natural tuning curveincreased with distance in túngara frogs. For the twosubspecies of cricket frogs, however, the relative performancedecreased at longer distances. The performance did not equalthe optimal tuning curve at the distances tested. The resultsindicate that the relationship between calls and auditory tuningcannot be optimal for both long and short distance reception.The relationship between female tuning and call dominant frequencymay represent a compromise between short and long distance communication,and the bias toward short or long distances may vary among species.     

Postembryonic ontogeny of the spadefoot toad,Scaphiopus intermontanus (Anura: Pelobatidae): Skeletal morphology

Postembryonic skeletal ontogeny of the pelobatid frog Scaphiopus intermontanus is described based on a developmental series of cleared-and-stained, whole-mount specimens. The focus is on laboratory-reared individuals fed a herbivorous diet as larvae. Although there is variation in the timing of ossification of individual skeletal elements relative to developmental stages based on external morphological criteria, the sequence of skeletal development generally is conservative. Compared with its close relative, S. bombifrons, ossifications that occur during prometamorphosis tend to be slightly delayed in S. intermontanus; however, cranial bones that ossify during late metamorphic climax in S. intermontanus are delayed until postmetamorphosis in S. bombifrons. The differences in timing between the two species are consistent, however, with differences observed between two developmental series of S. intermontanus raised at two different temperatures. Noteworthy features of skeletal development in S. intermontanus include: 1) presence of palatine ossifications that form from independent centers of ossification and soon fuse with the postnarial portion of the vomers to form the compound vomeropalatine bones; 2) compound sphenethmoid that may arise from four or more endochondral centers of ossification and one dorsal, dermal center of ossification; and 3) presence of transverse processes and vestigal prezygapophyses on the first postsacral vertebra. The morphology of the larval orbitohyoideus and interhyoideus muscles is compared. The record of skeletal ontogeny and muscle morphology presented herein for the herbivorous larval morph can serve as a baseline for comparisons with the ontogeny of the carnivorous larval morph of Scaphiopus. J. Morphol. 238:179–244, 1998. © 1998 Wiley-Liss, Inc.     

Responses of male cricket frogs (Acris crepitans) to attenuated and degraded advertisement calls

We examined the vocal and non‐vocal responses of male cricket frogs (Acris crepitans) to conspecific advertisement calls that had been attenuated or degraded by reducing the depth of amplitude modulation (AM). Both are characteristic of changes to the call as it is transmitted through natural habitats. As stimulus calls became more intense or less degraded, male cricket frogs gradually decreased their call rate and increased the number of call groups and pulse groups in their calls, changes indicative of increased aggressive interactions. At the higher intensities and lower degradation levels, the probability that males would shift to one of two non‐vocal behavioral responses, attacking the perceived intruder or ceasing calling and abandoning the call site, gradually increased. The results show that differences in signal attenuation and AM degradation levels are perceived by males and trigger both vocal and non‐vocal behavioral responses consistent with their use in evaluating the distance to a challenging male. Furthermore, the results indicate that the male responses are graded, increasing as intensity rises and degradation falls, and hierarchical, with vocal responses preceding behavioral responses over the range of intensities and degradation levels presented.     

Characterization,multiplex conditions,and cross‐species utility of tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi)

We have isolated and characterized 17 tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi), an anuran common in the central USA. Sixteen loci were organized into four multiplex amplification reactions. These loci were highly polymorphic when screened in 55 individuals from two distant populations, with 11–48 alleles per locus (average = 24.8). Observed and expected heterozygosities ranged from 0.18 to 0.97 and from 0.17 to 0.96, respectively. Nine loci were also polymorphic in Acris crepitans crepitans, with seven polymorphic in Acris gryllus. Five loci amplified in all three taxa. These loci will be useful for population‐ and species‐level investigations of this widespread group.     

Chondrocranium and cranial muscle morphology in Lysapsus and Pseudis tadpoles (Anura: Hylidae: Hylinae)

The chondrocranium and cranial muscles of Lysapsus limellum, Pseudis cardosoi, P. minuta and four subspecies of P. paradoxa (caribensis, occidentalis, paradoxa and platensis) are described. Lysapsus and Pseudis are related to the Hylidae by: (1) vertical processus anterolateralis of the larval crista parotica, and (2) presence of m. mandibulolabialis inferior and superior (the latter reversed in P. cardosoi). We confirm some larval synapomorphies previously proposed for the pseudids, except the m. subarcualis obliquus II having one head and the absence of copula anterior. We add three new features for the pseudids: (1) the distal end of ceratobranchiales III and IV projects towards either the capsula auditiva (Lysapsus) or the processus basicapsularis – new term – (Pseudis); (2) the arcus subocularis is twice as wide at the level of the processus ascendens as it is just posterior to the processus muscularis quadrati, and (3) the lateral corner of the processus articularis quadrati largely projects laterally. We diagnose Lysapsus larvae (the states of Pseudis in parentheses) as follows: (1) short processus pseudopterygoideus (long), (2) absence of lateral development of larval crista parotica (presence), (3) absence of commissura proximalis II (presence), (4) presence of processus lateralis hyalis – new term – (absence), (5) absence of processus basicapsularis (presence), (6) processus ascendens slightly curved along its entire extent (curved at the union with the basi cranii only), and (7) absence of m. tympanopharyngeus (presence).     

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.     

Skeletal development of the Mexican spadefoot, Spea multiplicata (Anura: Pelobatidae)

The larval chondrocranium of Spea multiplicata is described, as is the development and adult morphology of the skeleton. There are major modifications to the larval chondrocranium throughout development, including the presence of embryonic trabeculae in young tadpoles and significant reorganization of cartilaginous structures at metamorphosis. The first bone to ossify is the parasphenoid (Stage 35), followed by the presacral neural arches, ilium, and femur (Stage 36). By Stage 39, most of the postcranial elements have begun to ossify. Metamorphic climax is accomplished over three Gosner stages (39-41) and involves major modifications to the chondrocranium, as well as the appearance of three cranial elements (septomaxilla, nasal, and premaxilla). After metamorphosis, the exoccipital, vomer, dentary, angulosplenial, squamosal, pterygoid, sphenethmoid, ischium, and hyoid begin to ossify. The stapes, mentomeckelian, operculum, carpals, and tarsals do not appear until juvenile and adult stages. The development of the hyoid and cartilaginous condensations of the carpals and tarsals are described. In addition, phenotypic plasticity within the genus and the absence of a palatine (= neopalatine) bone are discussed.     

The ontogeny of Pseudis platensis (Anura,Hylidae): Heterochrony and the effects of larval development on postmetamorphic life

Recent studies have described the giant tadpole, delayed metamorphic transformations, and absence of postmetamorphic growth of the skeleton of Pseudis Platensis. These features address questions about derived patterns of life cycles and the role of the heterochrony during the metamorphosis in anurans. Using anatomical methods, we provide new data on the development of reproductive, digestive and integument systems, and age inference obtained from ontogenetic series of Pseudis platensis. Our results indicate that at the end of metamorphosis, the adult skin is completely differentiated, including the calcified dermal layer; the testis has seminiferous tubules with spermatogonia, spermatocytes, and spermatids; ovarian sacs present previtellogenic ova; and the adult digestive tract is fully formed. The froglets differ from adults only in being unable to reproduce. The entire life cycle of P. platensis can occur in 4 years. In the first year, larval development, growth to adult size, and gonad differentiation are completed. Long larval development rather than size of the tadpoles seems to be involved in the absence of juvenile stages. J. Morphol., 2010. © 2009 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.     

Phylogenetic relationships among marsupial frog genera (Anura: Hylidae: Hemiphractinae) based on evidence from morphology and natural history

We reassess the phylogenetic relationships of genera of hemiphractine hylid frogs (Marsupial Treefrogs) and discuss the evolution of several distinctive characters within this group using parsimony analysis. Fifty-one morphological and life-history characters were sampled from two species of Cryptobatrachus , three species of Flectonotus , 17 species of Gastrotheca , all five species of Hemiphractus , and one species of Stefania as the ingroup and three hyline, one phyllomedusine, and one pelodryadine species as outgroups. Our results support the mon-ophyly of Flectonotus, Cryptobatrachus , and Hemiphractus. Gastrotheca is paraphyletic with respect to Hemiphractus , dorsal pouches were lost in the ancestor of Hemiphractus. Direct development is a synapomorphy for Hemiphractinae and tadpoles were regained independently several times. These results stand in stark contrast to the prevailing paradigm regarding marsupial frog relationships.     

Morphology and development of additional bony elements in the genus Brachycephalus (Anura: Brachycephalidae)

We describe the extra bony elements, plates, and osteoderms present in species of the genus Brachycephalus. Samples of eight species of Brachycephalus, including seven populations of Brachycephalus ephippium, were examined. The large additional elements associated with the skull (parotic plate) and vertebrae (vertebral and paravertebral plates) all comprise intramembranous bone, similar to that of the frontoparietal or nasal bones of the skull of most of frogs. Additionally, in the dermis of one unnamed species, we discovered and described true osteoderms. We discuss the morphological nature and diversity of theses elements and their importance as evidence of phylogenetic relationship within Brachycephalus. In summary, three distinct conditions of extra bony elements occur in the genus Brachycephalus: (1) bony plates may be present or absent in species of the genus; (2) a few, small bony plates may be developed and these may be represented by (a) paravertebral plates small and restricted to the distal ends of the transverse processes of the presacral IV, (b) parotic plates small and not covering the tops of the squamosals, and (c) ornamented spinal plates on all vertebrae; and (3) well‐developed bony plates may be present as (a) paravertebral plates forming a ‘bone‐shield’ on the dorsal surface of the trunk, ornamented, and visible through the integument, (b) parotic plates covering the tops of the squamosals, and (c) spinal plates associated with all vertebrae, and ornamented on vertebrate I–VI. Although the phenomenon of miniaturization may be associated with the appearance of new elements in at least some of the species in the genus, the traditional rule may not be universally applicable. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 752–767.     

The larval hyobranchial apparatus of discoglossoid frogs: its structure and bearing on the systematics of the Anura (Amphibia: Anura)

The morphology of the larval hyobranchial apparatus of discoglossoid frog species representing the genera Ascaphus, Alytes, Bombina, and Discoglossus is described and the resulting characters were analysed cladistically. Seven species representing seven major lineages of frogs were included in the cladistic analysis of characters. Several changes in the terminology of the musculature are introduced, and a new interpretation of the subarcualis-muscle system is presented. The phylogenetic analysis suggest that the hyobranchial apparatus was substantially altered in the lineages leading to and within the Pipanura. This notably involved fusion, reduction and loss of skeletal structures and muscles, and splitting of certain muscles into muscle groups. The result confirm previous hypotheses based on the study of adults: discoglossoid species retain the most numlerous plesiomorphic characters among extant ianurans. The larval hyobranchial apparatus is in many features structrually similar to that of urodeles. Many of their character states were most likely present in the most recent common ancestor of all living forgs. The cladistic analysis of 31 characters of ithe larval hyobranchial apparatus supports major clades: Anura, Bombinanura, Pipanura, and Pelobatoidea + Neobatrachia. The cladiostic analysis and interpretation of larval characters is in part compatible with phylogenetic hypotheses based on characters of adults and rRNA sequences, but is in conflict with the Mesobatrachia and Archaeobatrachia concepts of other authors.     

How much fruit do fruit-eating frogs eat? An investigation on the diet of Xenohyla truncata (Lissamphibia: Anura: Hylidae)

This paper presents the results of a 22-month survey and the examination of the intestinal content of 356 specimens of Xenohyla truncata (Anura: Hylidae) from Restinga de Maricá, Rio de Janeiro, Brazil. Our results confirm prior observations that fruits are intrinsic to the diet of these frogs. In addition, these new data increase our understanding of the relationship between frogs and the plants they feed upon. Plant consumption follows availability of fruits in the area, indicating that the diversity of fruits consumed by the frogs does not represent choice, but rather plant phenology and fruit availability.