Robert C. Drewes

A molecular phylogenetic analysis of the family Rhacophoridae with an emphasis on the Asian and African genera

Using characters from mitochondrial DNA to construct maximum parsimony and maximum likelihood trees, we performed a phylogenetic analysis on representative species of 14 genera: 12 that belong to the treefrog family Rhacophoridae and two, Amolops and Rana, that are not rhacophorids. Our results support a phylogenetic hypothesis that depicts a monophyletic family Rhacophoridae. In this family, the Malagasy genera Aglyptodactylus, Boophis, Mantella, and Mantidactylus form a well-supported sister clade to all other rhacophorid genera, and Mantella is the sister taxon to Mantidactylus. Within the Asian/African genera, the genus Buergeria forms a well-supported clade of four species. The genera, except for Chirixalus, are generally monophyletic. An exception to this is that Polypedates dennysii clusters with species of Rhacophorus, suggesting that the taxonomy of the rhacophorids should be revised to reflect this relationship. Chirixalus is not monophyletic. Unexpectedly, there is strong support for Chirixalus doriae from Southeast Asia forming a clade with species of the African genus Chiromantis, suggesting that Chiromantis dispersed to Africa from Asia. Also, there is strong support for the sister taxon relationship of Chirixalus eiffingeri and Chirixalus idiootocus apart from other congeners.

Water, Nitrogen and Ion Balance in the African Treefrog Chiromantis petersi Boulenger (Anura: Rhacophoridae), With Comments on the Structure of the Integument

SummaryPhysiological and anatomical investigations were carried out onChiromantis petersi, an African rhacophorid treefrog, with the following results: 1.The minimum rate of evaporative water loss (EWL) was 0.41±0.25 mg/g.h.2.The maximum rate of water uptake in dehydrated frogs averaged 75% body weight/h in the first 10 min of rehydration.3.The low EWL correlates with the unique structure of the chromatophore units of the dorsum, the sides and the gular region.4.The high rates of water uptake correlate with the structure of the verrucae hydrophilica of the abdominal and femoral surfaces. These verrucae are not unique to this species.5.When denied water and force-fed mealworms for 30 days, plasma osmotic concentrations increased from 210 mosM to 384 mosM, with Na+, Cl− and urea contributing most to the increase.6.The ratio of urinary K+∶Na+ excretion is 3∶1.7.Excretion totaled 155 mg N/kg·day with 97% as uric acid, 2% as ammonia and 1% as urea.8.Since urea accumulated in the body at the rate of 58.5±6.1 mg N/kg·day, total nitrogen production was 213 mg N/kg·day. Uric acid formed 70% thereof.9.Chiromantis petersi is capable of surviving without free water for prolonged periods and is as well adapted to its xeric environment as are many desert reptiles.

Thermoregulatory Response to Heat in the Waterproof Frogs Phyllomedusa and Chiromantis

The thermal relations of waterproof frogs of two genera (Phyllomedusa and Chiromantis) were studied in an outdoor enclosure and, in the laboratory, in a thermal gradient, in a heated wind tunnel, and under an imposed radiant heat load. When allowed to move freely in a thermal gradient, no frogs showed a distinct preferred temperature, although Chiromantis spp. consistently avoided the cool end of the gradient. Both Chiromantis spp. and Phyllomedusa sauvagei voluntarily tolerated high body temperatures of 38 and 40 C, respectively. When subjected to a convective heat load, either outdoors or in the laboratory wind tunnel, both C. xerampelina and P. sauvagei allowed body temperature (Tb) to track air temperature (Ta) until Tb reached 38-39 C. Further increases in Ta resulted in little or no increase in Tb, whereas evaporative water loss (EWL) increased in direct proportion to the temperature difference (Ta − Tb) and with wind speed to the power of about 0.4. Phyllomedusa azurae increased water loss at a lower Tb (ca. 35-36 C) and did not regulate as precisely. A similar pattern was seen when frogs were subjected to rapid radiant heating. A sudden increase in EWL was observed when Chiromantis spp. reached a body temperature of 39 C and when P. azurae reached 35 C. Glands in the skin begin secretory activity when EWL increases, and the mechanism for thermoregulation in these frogs is apparently analogous to sweating.

Is The Amphibian Tree of Life really fatally flawed

Wiens (2007 , Q. Rev. Biol. 82, 55–56) recently published a severe critique of Frost et al.s (2006, Bull. Am. Mus. Nat. Hist. 297, 1–370) monographic study of amphibian systematics, concluding that it is “a disaster” and recommending that readers “simply ignore this study”. Beyond the hyperbole, Wiens raised four general objections that he regarded as “fatal flaws”: (1) the sampling design was insufficient for the generic changes made and taxonomic changes were made without including all type species; (2) the nuclear gene most commonly used in amphibian phylogenetics, RAG‐1, was not included, nor were the morphological characters that had justified the older taxonomy; (3) the analytical method employed is questionable because equally weighted parsimony “assumes that all characters are evolving at equal rates”; and (4) the results were at times “clearly erroneous”, as evidenced by the inferred non‐monophyly of marsupial frogs. In this paper we respond to these criticisms. In brief: (1) the study of Frost et al. did not exist in a vacuum and we discussed our evidence and evidence previously obtained by others that documented the non‐monophyletic taxa that we corrected. Beyond that, we agree that all type species should ideally be included, but inclusion of all potentially relevant type species is not feasible in a study of the magnitude of Frost et al. and we contend that this should not prevent progress in the formulation of phylogenetic hypotheses or their application outside of systematics. (2) Rhodopsin, a gene included by Frost et al. is the nuclear gene that is most commonly used in amphibian systematics, not RAG‐1. Regardless, ignoring a study because of the absence of a single locus strikes us as unsound practice. With respect to previously hypothesized morphological synapomorphies, Frost et al. provided a lengthy review of the published evidence for all groups, and this was used to inform taxonomic decisions. We noted that confirming and reconciling all morphological transformation series published among previous studies needed to be done, and we included evidence from the only published data set at that time to explicitly code morphological characters (including a number of traditionally applied synapomorphies from adult morphology) across the bulk of the diversity of amphibians (Haas, 2003, Cladistics 19, 23–90). Moreover, the phylogenetic results of the Frost et al. study were largely consistent with previous morphological and molecular studies and where they differed, this was discussed with reference to the weight of evidence. (3) The claim that equally weighted parsimony assumes that all characters are evolving at equal rates has been shown to be false in both analytical and simulation studies. (4) The claimed “strong support” for marsupial frog monophyly is questionable. Several studies have also found marsupial frogs to be non‐monophyletic. Wiens et al. (2005, Syst. Biol. 54, 719–748) recovered marsupial frogs as monophyletic, but that result was strongly supported only by Bayesian clade confidence values (which are known to overestimate support) and bootstrap support in his parsimony analysis was < 50%. Further, in a more recent parsimony analysis of an expanded data set that included RAG‐1 and the three traditional morphological synapomorphies of marsupial frogs, Wiens et al. (2006, Am. Nat. 168, 579–596) also found them to be non‐monophyletic. Although we attempted to apply the rule of monophyly to the naming of taxonomic groups, our phylogenetic results are largely consistent with conventional views even if not with the taxonomy current at the time of our writing. Most of our taxonomic changes addressed examples of non‐monophyly that had previously been known or suspected (e.g., the non‐monophyly of traditional Hyperoliidae, Microhylidae, Hemiphractinae, Leptodactylidae, Phrynobatrachus, Ranidae, Rana, Bufo; and the placement of Brachycephalus within “Eleutherodactylus”, and Lineatriton within “Pseudoeurycea”), and it is troubling that Wiens and others, as evidenced by recent publications, continue to perpetuate recognition of non‐monophyletic taxonomic groups that so profoundly misrepresent what is known about amphibian phylogeny.

Lymph Pools in the Basement, Sump Pumps in the Attic: The Anuran Dilemma for Lymph Movement

Amphibians are a vertebrate group transitional between aquatic and terrestrial environments. Consequently, both increases and decreases in blood volume are a natural biological stress associated with aquatic and terrestrial environments. In comparison with other vertebrate classes, anuran amphibians have the most rapid compensation and greatest capacity to compensate for changes in blood volume and survive dehydration. Unlike in mammals, a Starling transcapillary uptake mechanism does not account for this fluid mobilization because lymph flow is a substantial and important additional factor. The role of the lymphatic system in flux of fluids back into the circulation varies interspecifically in anurans and is an order of magnitude greater in anurans than in mammals. Current models of lymph movement in anurans are centered on the role of lymph hearts, but we suggest that these models are untenable. We present a new hypothesis for lymph movement involving (1) pressure differences created by compartmentalization of the hind limb lymph spaces into sacs of serially graded compliance to move lymph horizontally and (2) both negative and positive pressure differences created by contraction of skeletal muscles to move lymph vertically. The primary function of some of these skeletal muscles may be solely for lymph movement, but some may also be involved with other functions such as pulmonary ventilation.

Unique role of skeletal muscle contraction in vertical lymph movement in anurans

SUMMARY Electromyographic (EMG) activity of skeletal muscles that either insert on the skin or are associated with the margins of subcutaneous lymph sacs was monitored for two species of anurans, Chaunus marinus and Lithobates catesbeiana (formerly Bufo marinus and Rana catesbeiana). Our hypothesis was that contraction of these muscles varies the volume, and hence pressure, within these lymph sacs, and that this pressure is responsible for moving lymph from ventral, gravitationally dependent reaches of the body to dorsally located lymph hearts. EMG activity of M. piriformis, M. gracilis minor, M. abdominal crenator, M. tensor fasciae latae, M. sphincter ani cloacalis, M. cutaneous pectoris and M. cutaneous dorsi was synchronous with pressure changes in their associated lymph sacs. These muscles contracted synchronously, and the pressures generated within the lymph sacs were sufficient to move lymph vertically against gravity to the lymph hearts. The pressure relationships were complex; both negative and positive pressures were recorded during a contractile event, a pattern consistent with the addition and loss of lymphatic fluid to the lymph sacs. Severing the tendons of some of the muscles led to lymph pooling in gravitationally dependent lymph sacs. These data are the first to: (1) describe a function for many of these skeletal muscles; (2) document the role of skeletal muscles in vertical lymph movement in anurans; and (3) reinterpret the role of the urostyle, a bony element of the anuran pelvic girdle.

Reed frog diversification in the Gulf of Guinea: Overseas dispersal, the progression rule, and in situ speciation

Oceanic islands accumulate endemic species when new colonists diverge from source populations or by in situ diversification of resident island endemics. The relative importance of dispersal versus in situ speciation in generating diversity on islands varies with a number of archipelago characteristics including island size, age, and remoteness. Here, we characterize interisland dispersal and in situ speciation in frogs endemic to the Gulf of Guinea islands. Using mitochondrial sequence and genome‐wide single‐nucleotide polymorphism data, we demonstrate that dispersal proceeded from the younger island (São Tomé) to the older island (Príncipe) indicating that for organisms that disperse overseas on rafts, dispersal between islands may be determined by ocean currents and not island age. We find that dispersal between the islands is not ongoing, resulting in genotypically distinct but phenotypically similar lineages on the two islands. Finally, we demonstrate that in situ diversification on São Tomé Island likely proceeded in allopatry due to the geographic separation of breeding sites, resulting in phenotypically distinct species. We find evidence of hybridization between the species where their ranges are sympatric and the hybrid zone coincides with a transition from agricultural land to primary forest, indicating that anthropogenic development may have facilitated secondary contact between previously allopatric species.

Lymphatic regulation in nonmammalian vertebrates

All vertebrate animals share in common the production of lymph through net capillary filtration from their closed circulatory system into their tissues. The balance of forces responsible for net capillary filtration and lymph formation is described by the Starling equation, but additional factors such as vascular and interstitial compliance, which vary markedly among vertebrates, also have a significant impact on rates of lymph formation. Why vertebrates show extreme variability in rates of lymph formation and how nonmammalian vertebrates maintain plasma volume homeostasis is unclear. This gap hampers our understanding of the evolution of the lymphatic system and its interaction with the cardiovascular system. The evolutionary origin of the vertebrate lymphatic system is not clear, but recent advances suggest common developmental factors for lymphangiogenesis in teleost fishes, amphibians, and mammals with some significant changes in the water-land transition. The lymphatic system of anuran amphibians is characterized by large lymphatic sacs and two pairs of lymph hearts that return lymph into the venous circulation but no lymph vessels per se. The lymphatic systems of reptiles and some birds have lymph hearts, and both groups have extensive lymph vessels, but their functional role in both lymph movement and plasma volume homeostasis is almost completely unknown. The purpose of this review is to present an evolutionary perspective in how different vertebrates have solved the common problem of the inevitable formation of lymph from their closed circulatory systems and to point out the many gaps in our knowledge of this evolutionary progression.

Physiological vagility and its relationship to dispersal and neutral genetic heterogeneity in vertebrates

Vagility is the inherent power of movement by individuals. Vagility and the available duration of movement determine the dispersal distance individuals can move to interbreed, which affects the fine-scale genetic structure of vertebrate populations. Vagility and variation in population genetic structure are normally explained by geographic variation and not by the inherent power of movement by individuals. We present a new, quantitative definition for physiological vagility that incorporates aerobic capacity, body size, body temperature and the metabolic cost of transport, variables that are independent of the physical environment. Physiological vagility is the speed at which an animal can move sustainably based on these parameters. This meta-analysis tests whether this definition of physiological vagility correlates with empirical data for maximal dispersal distances and measured microsatellite genetic differentiation with distance {[FST/[1−FST)]/ln distance} for amphibians, reptiles, birds and mammals utilizing three locomotor modes (running, flying, swimming). Maximal dispersal distance and physiological vagility increased with body mass for amphibians, reptiles and mammals utilizing terrestrial movement. The relative slopes of these relationships indicate that larger individuals require longer movement durations to achieve maximal dispersal distances. Both physiological vagility and maximal dispersal distance were independent of body mass for flying vertebrates. Genetic differentiation with distance was greatest for terrestrial locomotion, with amphibians showing the greatest mean and variance in differentiation. Flying birds, flying mammals and swimming marine mammals showed the least differentiation. Mean physiological vagility of different groups (class and locomotor mode) accounted for 98% of the mean variation in genetic differentiation with distance in each group. Genetic differentiation with distance was not related to body mass. The physiological capacity for movement (physiological vagility) quantitatively predicts genetic isolation by distance in the vertebrates examined.

Physiological Vagility: Correlations with Dispersal and Population Genetic Structure of Amphibians*

Physiological vagility represents the capacity to move sustainably and is central to fully explaining the processes involved in creating fine-scale genetic structure of amphibian populations, because movement (vagility) and the duration of movement determine the dispersal distance individuals can move to interbreed. The tendency for amphibians to maintain genetic differentiation over relatively short distances (isolation by distance) has been attributed to their limited dispersal capacity (low vagility) compared with other vertebrates. Earlier studies analyzing genetic isolation and population differentiation with distance treat all amphibians as equally vagile and attempt to explain genetic differentiation only in terms of physical environmental characteristics. We introduce a new quantitative metric for vagility that incorporates aerobic capacity, body size, body temperature, and the cost of transport and is independent of the physical characteristics of the environment. We test our metric for vagility with data for dispersal distance and body mass in amphibians and correlate vagility with data for genetic differentiation (). Both dispersal distance and vagility increase with body size. Differentiation () of neutral microsatellite markers with distance was inversely and significantly () related to ln vagility. Genetic differentiation with distance was not significantly related to body mass alone. Generalized observations are validated with several specific amphibian studies. These results suggest that interspecific differences in physiological capacity for movement (vagility) can contribute to genetic differentiation and metapopulation structure in amphibians.