Hydric constraints upon foraging in a terrestrial salamander,Desmognathus ochrophaeus (Amphibia: Plethodontidae)

Summary The time until salamanders voluntarily abandoned foraging (the water time limit) and the amount of water lost when salamanders abandoned foraging (dehydration deficit) were determined for terrestrial plethodontid salamanders, Desmognathus ochrophaeus, foraging at various vapor pressure gradients in the laboratory. Salamander activity was correlated with the rate of water loss and was inversely related to the water time limit. Animals at 0.35–0.86 kPa vapor pressure gradients abandoned foraging and returned to moist retreats significantly sooner than animals in water-saturated air. The early retreat of animals in dry air was related in part to high rates of water loss and in part to the modest dehydration deficit (3.8%) at which animals abandoned foraging. Locomotor performance and foraging ability were unaffected by dehydration until dehydration deficits exceeded 12%. This suggests that salamanders in unsaturated air abandoned foraging at a low dehydration deficit to conserve and replenish water reserves rather than to avoid outright incapacitation or death. Present address: Department of Zoology, University of Guelph, Guelph, Ontario N1G 2W1, Canada     

Population differentiation in plethodontid salamanders: divergence of allozymes and sexual compatibility among populations of Desmognathus imitator and D. ochrophaeus (Gaudata: Plethodontidae)

Plethodontid salamanders of the genus Desmognathus exhibit varying levels of genetic differentiation among and within both allopatric and sympatric taxa. This provides excellent opportunities to study population differentiation and speciation. Two morphologically similar species in this genus, D. imitator and D. ochrophaeus, are genetically well-differentiated from one another and occur in sympatry with no evidence of hybridization and introgression. We report that the degree of sexual isolation between these two species is very high, regardless of whether the populations under comparison are allopatric or sympatric with one another. Neither reinforcement nor reproductive character displacement are required to explain the evolution of sexual incompatibility in sympatry. Sexual behaviour apparently diverges while populations are allopatric with one another. Preliminary study indicates that D. imitator consists of populations among which there may be significant sexual isolation in the absence of detectable genetic differentiation.     

The courtship behaviour of the spotted dusky salamander, Desmognathus fuscus conanti (Amphibia: Caudata: Plethodontidae)

The courtship behaviour of the North American southern dusky salamander, Desmognathus fuscus conanti (Plethodontidae), is described for the first time. The male is the most active participant, as is typical for caudate amphibians, and much of his behaviour can be understood in terms of orientation to and stimulation (or persuasion) of the female. The latter is largely accomplished by the transfer of courtship pheromones from the male's mental (chin) gland. The orientation and persuasion phases of courtship appear necessary if sperm transfer by means of a spermatophore deposited upon the substratum is to be successful. The courtship of D. fuscus conanti is qualitatively identical to that of D. santeetlah , with which it is sexually compatible in the laboratory and with which it may hybridize in nature. However, a broader comparison of species in the genus Desmognathus reveals that the greatest interspecific differences occur during the orientation and persuasion phases of courtship. A complete understanding of the evolution of courtship behaviour in desmognathine salamanders awaits availability of ethograms for all other taxa and a robust, independent phytogeny for the entire subfamily.     

Osseous pathologies in the lungless salamander Desmognathus fuscus (Plethodontidae)

The majority of reported pathologies in lissamphibians (salamanders, caecilians and frogs) include limb deformities such as missing limbs, multiple extra limbs and digits, or incomplete limb formation. However, comparatively little is known about congenital vertebral malformations or posttraumatic pathologies (e.g. injuries, infections) in the vertebral column of salamanders. In the present study, we describe eight vertebral deformities in three cleared and stained specimens of Desmognathus fuscus. Two specimens display developmental deformities which range from a potential non-segmented wedge vertebra to fully segmented hemivertebrae. The vertebral pathology in the third specimens possibly results from a parasitic infection. Apparently, these osseous deformities were not severe enough to prohibit survival of the specimens.     

Divergence of mate recognition in the African striped mouse (Rhabdomys)

Mate recognition systems (MRSs) are complex signal-receivertraits. The present study addressed the roles of phylogeny,ecology, and geography in shaping the MRS of the African stripedmouse (Rhabdomys), which has a wide distribution in southernAfrica. Two putative species are recognized, which have differentecologies: Rhabdomys pumilio (arid) and Rhabdomys dilectus (mesic).The latter may be further subdivided into 2 subspecies (R. dilectusdilectus and R. dilectus chakae). Using 2 discrete populationsper taxon, we investigated within- and between-taxon variationsin male odor quality and female perception using habituation-discriminationand habituation-generalization tests, and female preferencein 2-way choice tests. Our results indicate: 1) no within-taxonvariation in odor quality, perception, or preference; 2) the2 subspecies of R. dilectus carry signals of different qualitiesbut share a common odor characteristic distinct from that ofR. pumilio; 3) female R. pumilio did not show a preference whentheir own species and R. d. chakae odors were presented simultaneouslybut displayed assortative preference when the alternative wasR. d. dilectus; 4) females of the 2 subspecies showed dissimilarpreferences: R. d. chakae for the genetically more similar taxonand R. d. dilectus for the most different one. Although we couldnot rule out the influence of ecology, we concluded that phylogenyappeared a more parsimonious explanation for the pattern ofdivergence in Rhabdomys. Further, we discuss our results inlight of current models on signal-receiver coevolution.     

Histology of mental and caudal courtship glands in three genera of plethodontid salamanders (Amphibia: Plethodontidae)

Salamanders in the family Plethodontidae exhibit a unique tail‐straddle walk during courtship that can include the use of sexually dimorphic mental and caudal courtship glands. This study presents novel histological and fine structure data on mental glands and caudal courtship glands in Plethodon mississippi, Desmognathus conanti and Eurycea quadridigitata using both light microscopy and scanning electron microscopy. This study represents the first use of scanning electron microscopy to observe these glands. Both mental and caudal courtship glands were observed to vary seasonally in gland diameter and histology according to the breeding season of each species. Morphological variation was observed across the three species studied in both clustering and relative size of the glands compared to neighbouring mucous and granular glands. Hypertrophied mental glands are larger than mucous or granular glands in all species, but relationships among caudal courtship glands and other skin glands vary among species. In E. quadridigitata, active caudal courtship glands are larger than mucous and granular glands, but in D. conanti, caudal courtship glands are similar in size to granular glands and larger than mucous glands. In P. mississippi, caudal courtship glands are scattered among significantly larger granular glands and are similar in size to mucous glands.     

Sperm storage in the spermatheca of the red-back salamander,Plethodon cinereus (Amphibia: Plethodontidae)

In northern Indiana, the mating season of Plethodon cinereus occurs after hibernation from March until June, when oviposition begins. During the mating season, a female stores sperm in its spermatheca, a compound tubular gland in the roof of the cloaca. The apical cytoplasm of the spermathecal epithelium is filled with large secretory vacuoles whose product is released while sperm are stored. Females induced to oviposit in June and July by injections of human chorionic gonadotropin (hCG) still retain much sperm 1 month after oviposition, but secretory vacuoles are absent in all specimens sacrificed in July and August. Instead, some sperm are embedded in the spermathecal epithelium with resultant spermiophagy involving lysosomes. A female sacrificed in September 2 months after oviposition possesses scant sperm, but spermiophagy alone does not seem extensive enough to account for the decrease in sperm numbers. Females sacrificed in October prior to hibernation lack sperm in their spermathecae; some secretory vacuoles are present, but they are not as numerous or as enlarged as in specimens collected in March and May. Inter- and intrafamilial differences in the cytology of sperm storage may not be phyletically informative at the family level but related to species-specific reproductive adaptations. J. Morphol. 234:131–146, 1997. © 1997 Wiley-Liss, Inc.     

Recognition by Population and Genetic Similarity in the Mountain Dusky Salamander,Desmognathus ochrophaeus (Amphibia: Plethodontidae)

Adult males and females of Desmognathus ochrophaeus were tested in two-choice substrate discrimination trials in which salamanders were given a choice of the marked substrates of opposite-sex individuals from their own population and from a geographically distinct population. Females exhibited a significant preference for the marked substrates of extrapopulation males, whereas there was no evidence that males made any such discrimination of the marked substrates of females. We suggest that females may utilize substrate cues to make initial assessments of potential mates. We compared data from substrate discrimination trials to random amplified polymorphic DNA (RAPD) profiles generated for each individual. There was no evidence that females made any distinction of substrate odours based on our assessment of genetic similarity. The estimated genetic differences between the two populations were minimal.     

Complex secretory products in the oviduct of the plethodontid salamander Bolitoglossa dofleini (Amphibia, Urodela)

The ultrastructure of the secretory granules in the cells of the subdivisions of the oviduct in the neotropical plethodontid salamander Bolitoglossa dofleini was studied by transmission electron microscopy. In addition, we applied the cationic dye Cuprolinic Blue (CB) at different electrolyte concentrations to demonstrate proteoglycans, and the pyrogallol red-copper (PR-C) method to stain proteins at the ultrastructural level. The entire oviduct is lined by a simple epithelium that contains ciliated and microvillous cells in the first subdivision, the aglandular pars recta; microvillous cells show a moderate secretory activity. The following pars convoluta is differentiated into five glandular subdivisions and the aglandular “uterine portion”. Especially in the glandular parts, the epithelium is arranged in longitudinal folds. At their crests ciliated and microvillous cells similar to those in the pars recta occur. Gland cells are crowded with secretory granules that differ in their structural complexity (with and without electron-dense spheres or masses; elaborated, homogeneous or granular matrix; spherical; distorted) along the various subdivisions. Further, as suggested by the CB-technique, the cranial subdivisions contain large amounts of sulphated proteoglycans that decrease in the caudal direction. Carboxylated proteglycans appear to be present in all subdivisions examined. Electron-dense spheres of secretory granules are largely free of CB-precipitates, but stain more or less intensely with PR-C. The ultrastructure of the pars recta, and especially the “uterine portion” indicates transporting capability. The epithelial cells of the “uterus” have coated pits and a considerable amount of lysosome-like bodies.     

Heterochronic mechanisms of morphological diversification and evolutionary change in the neotropical salamander,Bolitoglossa occidentalis (Amphibia: Plethodontidae)

Bolitoglossa occidentalis, a lowland salamander of Mexico and Guatemala, has a highly derived morphology. The features that are derived with respect to the condition in generalized members of the genus include the following: (1) small body size; (2) short tail; (3) fully webbed hands and feet; (4) reduction and loss of certain phalangeal elements; (5) fusion of carpals and tarsals; (6) absence of prefrontal bones; and (7) reduced skull ossification. The ontogeny of this species was analyzed quantitatively and compared with the patterns of growth and differentiation encountered in two morphologically generalized members of the genus, B. rostrata and B. subpalmata. Most of the derived features can be explained by invoking a single heterochronic process: truncation of development at a small size (most likely the product of early maturation). Therefore, B. occidentalis is a paedomorphic species whose morphology has been attained through the process of progenesis. This result supports Alberch's ('80a) prediction, based on functional analysis, that the principle adaptation to arboreality in B. occidentalis is small size; other derived morphological features are associated with the organism's truncated development and may have no adaptive significance. However, patterns of dissociation are found within this overall progenetic process. Some of these include the following: (1) accelerated growth rates of the metatarsals and first phalanges, and retarded growth rates of the second and third phalangeal elements; (2) dissociation between rates of ossification of the skull and the autopodial elements; and (3) dissociation between the timing of termination of the process of shape change during the ontogeny of the foot (the product of differential growth between digital and interdigital areas) and termination of growth in overall foot size (foot surface area). This later result illustrates the independence of morphogenetic phenomena (shape change) from processes of growth (size increase).     

Productivity and significance of headwater streams: population structure and biomass of the black-bellied salamander (Desmognathus quadramaculatus)

1. Headwater streams are a significant feature of the southern Appalachian landscape, comprising more than 70% of the total stream length in the region. Salamanders are the dominant vertebrate within headwater‐riparian forest ecosystems, but their ecological role is not clearly understood. 2. We studied a population of black‐bellied salamanders (Desmognathus quadramaculatus) at a headwater stream in the southern Appalachian Mountains using radio‐telemetry and mark‐recapture methods. The length and area of headwater streams in the region were estimated using GIS. 3. Home ranges of radio‐tracked salamanders were relatively small (mean = 1.06 m²). Adult salamanders in our telemetry study inhabited edge microhabitats significantly more often than either stream or riparian microhabitats, and the same trend was observed in the mark‐recapture study. 4. We estimated the population density at this site to be 11 294 salamanders ha^?1, amounting to 99.30 kg ha^?1 of biomass, an estimate that is six times greater than reported in previous studies. The majority of this biomass was found within the stream, but 22% was found in the surrounding riparian habitat more than 1 m from the stream. Using headwater stream length and area estimates, we extrapolated biomass estimates for black‐bellied salamanders inhabiting stream and riparian microhabitats across the study region. 5. We report one of the largest estimates of secondary consumer biomass for a headwater ecosystem, attesting to the overall productivity of headwater streams. Headwaters are known to be important for ecological and ecosystem processes and our biomass estimates suggest that salamanders are a critical component to these systems.     

Environmental factors associated with genetic and phenotypic divergence among sympatric populations of Arctic charr (Salvelinus alpinus)

The mechanisms by which phenotypic and genetic divergence may occur among sympatric, conspecific populations have been widely discussed but are still not well understood. Possible mechanisms include assortative mating based on morphology or variation in the reproductive behaviour of phenotypes, and both have been suggested to be relevant to the differentiation of salmonid populations in post-glacial lakes. Here, we studied Arctic charr (Salvelinus alpinus) in Windermere, where putative populations are defined by spatial and temporal variation in spawning. Genetic differentiation was assessed based on nine microsatellite loci, and phenotypic variation was assessed from morphometric characters. We test hypotheses about the relative role of morphology, spawning season and spawning habitat in the evolution of genetic divergence among these populations. Distinct from other lake systems, we find that both morphological and genetic differentiation are restricted primarily to one of two interconnecting basins, that genetic and morphological differentiation are decoupled in this lake and that both phenotype and environment have changed over the last 20 years. The implication is that breeding habitat plays a primary role in isolating populations that differentiate by drift and that phenotypically plastic changes, potentially related to foraging specializations, have either become secondarily decoupled from the genetically defined populations or were never fundamental in driving the evolution of genetic diversity in the Windermere system.     

Geographic allozyme variation in the Japanese clouded salamander, Hynobius nebulosus (Amphibia: Urodela)

Using starch gel electrophoresis, geographical enzyme variation was surveyed in 46 populations of the Japanese salamander Hynobius nebulosus , which occurs widely in western Japan. This species exhibits substantial local genetic differentiation and is more diverse in inland regions than on small islands. In several different analyses, two groups of populations, one from Kyushu Island to the westernmost part of Chugoku district, Honshu (western group), and another from Shikoku through Kinki to Chubu district, Honshu (eastern group), were consistently recognized. Most of the remaining populations were placed in two less clearly-defined groups: the montane group from the Chugoku Mountains and the Chugoku group from the coastal regions of Chugoku district. The divergence patterns of H. nebulosus are thought to be related to the geological history of its range. The populations near the boundary of the western group were found to share several alleles that were predominant on both sides of the boundary, indicating past secondary contact. Differentiation in the montane group did not follow the isolation by distance model, probably because several populations included were genetically mixed with lowland populations of the Chugoku group.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 311–330.     

Structural heterogeneity in the basal regions of the teeth of the red-backed salamander,Plethodon cinereus (Amphibia,Plethodontidae)

Ultrastructural and histochemical features of marginal (monostichous) teeth associated with the jawbones are compared with those of palatal (polystichous) teeth that compose two patches in the roof of the mouth. The apices and uncalcified regions are similar in both kinds of teeth, but the basal regions display distinctive differences. While bases (pedestals) of marginal teeth are essentially hollow cylinders that attach to the jawbones by their labial faces, bases of teeth in palatal patches are fused to form two horizontal plates which lack direct attachment to underlying bone. The plates are separated from each other by a pulp-filled space containing fibroblasts, blood vessels, and vertically oriented elements resembling bony spicules. Cylindrical pedestals like those of marginal teeth project from the ventral plate. While the identity of the material composing the basal regions remains controversial, the following evidence suggests that it is similar to “bone of attachment” (Tomes, '23): most of it, unlike dentin, does not develop in direct association with an enamel organ; alcian blue stains the bases of developing teeth but stains dentin, developing dentin, enamel, or mature bone very weakly (if at all); bases of teeth in palatal patches develop in isolation from the parasphenoid bone and thus cannot be considered extensions of it; and marginal teeth attach directly to the jawbones, but the material composing their bases does not blend with the bone. Structural heterogeneity of the basal regions appears to be linked to functional differences exhibited by these two types of teeth.     

Histology and ultrastructure of the caudal courtship glands of the red‐backed salamander,Plethodon cinereus (Amphibia: Plethodontidae)

Caudal courtship glands (CCGs) are sexually dimorphic glands described in the skin of the dorsal tail base of some male salamanders in the genera Desmognathus, Eurycea, and Plethodon in the family Plethodontidae. These glands are believed to deliver pheromones to females during courtship, when the female rests her chin on the dorsal tail base during the stereotypic tail straddling walk unique to plethodontids. Although CCGs have been studied histologically, no investigations of their ultrastructure have been made. This article presents the first study on the fine structure and seasonal variation of CCGs, using the plethodontid Plethodon cinereus. The CCGs vary seasonally in height and secretory activity. The mature secretory granules observed in males collected in October and April consist of oval, biphasic granules that are eosinophilic and give positive reactions to periodic acid‐Schiff for neutral carbohydrates but do not stain for acidic mucosusbtances or proteins with alcian blue and bromphenol blue, respectively. Granular glands, some of which contain mucous demilunes, are twice as large as CCGs, are syncytial (unlike CCGs), and stain for proteins. Mucous glands are similar in size to CCGs, but are basophilic, show no seasonal variation in secretory activity, and stain positive for acidic mucosubstances. CCGs do not resemble cytologically the sexually dimorphic mental glands of some plethodontids, which contain round or oval granules filled with an electron‐dense amorphous substance. The CCGs are similar histologically to sexually dimorphic skin glands described in some anurans, but more comparative work is needed. J. Morphol. 276:319–330, 2015. © 2014 Wiley Periodicals, Inc.     

Structure of the radially asymmetrical uncalcified region of the teeth of the red-backed salamander,Plethodon cinereus (Amphibia,Plethodontidae)

The teeth of the adult plethodontid salamander, Plethodon cinereus, were examined by light and electron microscopy with emphasis on the ringlike zone of uncalcified dentin that divides the calcified portion of each tooth into a proximal pedestal and a distal apex. The uncalcified region displays radial asymmetry, forming an integral part of the posterior wall of the tooth but bulging into the pulp cavity anteriorly, thus forming a hingelike structure. All portions of the dentin, including the uncalcified region, are composed predominantly of collagenous fibers but lack elastin. In scanning electron micrographs of teeth from which the oral mucosa has been removed, the location of the anterior uncalcified hinge is marked externally by a notch-like articulation of the apex and pedestal. Sites of transition between calcified and uncalcified areas of the dentin show no special modifications in transmission electron micrographs, but collagenous fibers in calcified portions are associated with more electron-dense amorphous material than are those in the uncalcified region. Odontoblasts associated with the uncalcified region possess ultrastructural features closely resembling those of odontoblasts found in calcified areas. The uncalcified region seems to afford the teeth a certain degree of flexibility, and the asymmetry of the region appears to allow the teeth to flex only in a posterior direction, thus facilitating the entry of living prey but hindering its escape. The uncalcified region also seems to permit the apex of a tooth to break away from its pedestal without damage to underlying bone.