Characterization of nine polymorphic microsatellite loci in the dyeing poison frog Dendrobates tinctorius (Dendrobatidae), and their cross-species utility in two other dendrobatoid species

While field and laboratory based studies have provided significant insights into the parental care and courtship behaviour of dendrobatoid frogs, a comprehensive assessment of their genetic mating systems and population genetic parameters has been precluded because ofthe lack of highly variable DNA markers. Here we document the development of nine novel polymorphic microsatellite markers for the dyeing poison frog Dendrobates tinct or ius (Dendrobatidae ). We found between three and 16 alleles per locus in 60 individuals (30 males, 30 females) from the field site Saut Parare, French Guiana, with an average observed heterozygosity of 0. 75. None of the loci deviated significantly from Hardy-Weinberg equilibrium or showed linkage disequilibrium. We also report successful cross-species amplification of the nine markers in two other dendrobatoid species (Allobates femora/is and Oophaga pumilio). These markers have the potential to aid in determining the genetic structure of local populations, identifying small-scale phylogenies such as parent-offspring relationships and will allow for cross-species comparisons within dendrobatoid species. Therefore, these markers can be applied to a wide range of scientific fields, such as conservation, behavioural ecology and evolutionary biology.

Intraspecific and intraspecific views of colour signals in the strawberry poison frog Dendrobates Pumilio

Poison frogs in the anuran family Dendrobatidae use bright colors on their bodies to advertise toxicity. The species Dendrobates pumilio Schmidt 1858, the strawberry poison frog, shows extreme polymorphism in color and pattern in Panama. It is known that females of D. pumilio preferentially choose mates of their own color morph. Nevertheless, potential predators must clearly see and recognize all color morphs if the aposermatic signaling system is to function effectively. We examined the ability of conspecifics and a model predator to discriminate a diverse selection of D. pumilio colors from each other and from background colors. Microspectrophotometry of isolated rod and cone photoreceptors of D. pumilio revealed the presence of a trichromatic photopic visual system. A typical tetrachromatic bird system was used for the model predator. Reflectance spectra of frog and background colors were obtained, and discrimination among spectra in natural illuminants was mathematically modeled. The results revealed that both D. pumilio and the model predator discriminate most colors quite well, both from each other and from typical backgrounds, with the predator generally performing somewhat better than the conspecifics. Each color morph displayed at least one color signal that is highly visible against backgrounds to both visual systems. Our results indicate that the colors displayed by the various color morphs of D. pumilio are effective signals both to conspecifics and to a model predator.

Ontogenetic Scaling of Poison Glands in Dendrobates Pumilio

The relationship between granular (poison) gland size and density was examined in an ontogenetic series of the strawberry dart-poison frog, Dendrobates pumilio. Specimens used in this study were collected from the La Selva Biological Station in northeastern Costa Rica. Patches of skin from the dorsal surface of seven frogs, ranging in size from 11 to 23 mm snout-vent length (SVL), were fixed and embedded in paraffin for histological sectioning. Poison gland size and density were quantified microscopically in these sections. Poison glands are uniformly distributed across the skin and mean poison gland diameter increases at a rate faster than snout-vent length from 42.5 at SVL 11mm to 120.0 at SVL 23 mm. Conversely, gland density decreases with body size from 71.9 glands/mm2 to 33.2 glands/mm2 • Due to the positive allometric growth of the poison glands, the percentage of skin surface occupied by poison glands increases from 10.1-22.1% in small frogs (SVL<18 >mm) to 50.0-65.2% in large frogs (SVL>19MM), resulting in more toxin per mm2 in the larger animals. The largest increase in toxicity is correlated temporally with the onset of sexual maturity rather than with changes in aposematic coloring.

高原鼠兔种群的性比

性比是反映动物种群的基本特征之一，因此成为种群生态学的主要研究内容。就种群的性比而言，在不同的物种、或同一物种在不同的时期和条件下都有极大的不同~[1-4]。性比的改变将影响动物的种群结构、家庭组成和交配关系，也影响动物两性的配偶竞争、繁殖投入、繁殖成功以及性二型的分化等~[4-13]。在繁殖过程中，动物可以采用改变性比的方法调节种群的密度和大小[2,14,15]。同时，动物的性比受多种因素的影响，不仅存在季节、年间以及地理变化的差异，而且受环境条件(如气温和降水量)的影响~[2～4]。捕食风险也是影响性比的主要因素。虽然许多动物出生时的性比是一样的，但是，由于捕食者在捕食猎物的过程中存在明显的选择性，使其性比发生了明显的变化~[16]。对于动物种群性比的研究已有诸多报道，其研究的动物包括海蛇尾(Ophiactis savignyi)~[5]、龙虾(Homarus gam- marus)~[6]、射毒蛙(Dendrobates purnilio)~[9]、斑马雀(Taeniopygyia guttata castanotis)~[8]、褐家鼠(Rattus norvegicus)~[17]等。高原鼠兔(Ochotona curzoniae)是青藏高原草地生态系统中的优势小哺乳类动物。有关高原鼠兔种群性比的研究，国内学者有过涉及~[18,19]，但所作工作多是在高原鼠兔繁殖生态学研究中进行的静态描述，对种群性比的年龄变化，季节和年度变化及其与环境条件的关系等方面的研究尚缺详细的报道。本文通过对高原鼠兔种群性比的年间比较、不同胎次性比的分析和越冬对其性比影响的研究，旨在探讨高原鼠兔的性比变化在其种群调节中的作用。

Paradox lost: variable colour-pattern geometry is associated with differences in movement in aposematic frogs

Aposematic signal variation is a paradox: predators are better at learning and retaining the association between conspicuousness and unprofitability when signal variation is low. Movement patterns and variable colour patterns are linked in non-aposematic species: striped patterns generate illusions of altered speed and direction when moving linearly, affecting predators' tracking ability; blotched patterns benefit instead from unpredictable pauses and random movement. We tested whether the extensive colour-pattern variation in an aposematic frog is linked to movement, and found that individuals moving directionally and faster have more elongated patterns than individuals moving randomly and slowly. This may help explain the paradox of polymorphic aposematism: variable warning signals may reduce protection, but predator defence might still be effective if specific behaviours are tuned to specific signals. The interacting effects of behavioural and morphological traits may be a key to the evolution of warning signals. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Convergent evolution of aposematic coloration in Neotropical poison frogs: a molecular phylogenetic perspective

Poison frogs of the family Dendrobatidae contain cryptic as well as brightly colored, presumably aposematic species. The prevailing phylogenetic hypothesis assumes that the aposematic taxa form a monophyletic group while the cryptic species (Colostethus sensu lato) are basal and paraphyletic. Analysis of 86 dendrobatid sequences of a fragment of the 16S rRNA gene resulted in a much more complex scenario, with several clades that contained aposematic as well as cryptic taxa. Monophyly of the aposematic taxa was significantly rejected by SH-tests in an analysis with additional 12S and 16S rDNA fragments and reduced taxon sampling. The brightly colored Allobates femoralis and A. zaparo (Silverstone) comb. nov. (previously Epipedobates) belong in a clade with cryptic species of Colostethus. Additionally, Colostethus pratti was grouped with Epipedobates, and Colostethus bocagei with Cryptophyllobates. In several cases, the aposematic species have general distributions similar to those of their non-aposematic sister groups, indicating multiple instances of regional radiations in which some taxa independently acquired bright color. From a classificatory point of view, it is relevant that the type species of Minyobates, M. steyermarki, resulted as the sister group of the genus Dendrobates, and that species of Mannophryne and Nephelobates formed monophyletic clades, corroborating the validity of these genera. Leptodactylids of the genera Hylodes and Crossodactylus were not unambiguously identified as the sister group of the Dendrobatidae; these were monophyletic in all analyses and probably originated early in the radiation of Neotropical hyloid frogs.

Phylogenetic systematics of dart-poison frogs and their relatives (Amphibia: Athesphatanura: Dendrobatidae)

The known diversity of dart-poison frog species has grown from 70 in the 1960s to 247 at present, with no sign that the discovery of new species will wane in the foreseeable future. Although this growth in knowledge of the diversity of this group has been accompanied by detailed investigations of many aspects of the biology of dendrobatids, their phylogenetic relationships remain poorly understood. This study was designed to test hypotheses of dendrobatid diversification by combining new and prior genotypic and phenotypic evidence in a total evidence analysis. DNA sequences were sampled for five mitochondrial and six nuclear loci (approximately 6,100 base pairs [bp]; x=3,740 bp per terminal; total dataset composed of approximately 1.55 million bp), and 174 phenotypic characters were scored from adult and larval morphology, alkaloid profiles, and behavior. These data were combined with relevant published DNA sequences. Ingroup sampling targeted several previously unsampled species, including Aromobates nocturnus, which was hypothesized previously to be the sister of all other dendrobatids. Undescribed and problematic species were sampled from multiple localities when possible. The final dataset consisted of 414 terminals: 367 ingroup terminals of 156 species and 47 outgroup terminals of 46 species. Direct optimization parsimony analysis of the equally weighted evidence resulted in 25,872 optimal trees. Forty nodes collapse in the strict consensus, with all conflict restricted to conspecific terminals. Dendrobatids were recovered as monophyletic, and their sister group consisted of Crossodactylus, Hylodes, and Megaelosia, recognized herein as Hylodidae. Among outgroup taxa, Centrolenidae was found to be the sister group of all athesphatanurans except Hylidae, Leptodactyidae was polyphyletic, Thoropa was nested within Cycloramphidae, and Ceratophryinae was paraphyletic with respect to Telmatobiinae. Among dendrobatids, the monophyly and content of Mannophryne and Phyllobates were corroborated. Aromobates nocturnus and Colostethus saltuensis were found to be nested within Nephelobates, and Minyobates was paraphyletic and nested within Dendrobates. Colostethus was shown to be rampantly nonmonophyletic, with most species falling into two unrelated cis- and trans-Andean clades. A morphologically and behaviorally diverse clade of median lingual process-possessing species was discovered. In light of these findings and the growth in knowledge of the diversity of this large clade over the past 40 years, we propose a new, monophyletic taxonomy for dendrobatids, recognizing the inclusive clade as a superfamily (Dendrobatoidea) composed of two families (one of which is new), six subfamilies (three new), and 16 genera (four new). Although poisonous frogs did not form a monophyletic group, the three poisonous lineages are all confined to the revised family Dendrobatidae, in keeping with the traditional application of this name. We also propose changes to achieve a monophyletic higher-level taxonomy for the athesphatanuran outgroup taxa. Analysis of character evolution revealed multiple origins of phytotelm-breeding, parental provisioning of nutritive oocytes for larval consumption (larval oophagy), and endotrophy. Available evidence indicates that transport of tadpoles on the dorsum of parent nurse frogs-a dendrobatid synapomorphy-is carried out primitively by male nurse frogs, with three independent origins of female transport and five independent origins of biparental transport. Reproductive amplexus is optimally explained as having been lost in the most recent common ancestor of Dendrobatoidea, with cephalic amplexus arising independently three times. © American Museum of Natural History 2006.

Batrachochytrium dendrobatidis in amphibians confiscated from illegal wildlife trade and used in an ex situ breeding program in Brazil

This paper describes an outbreak of chytridiomycosis affecting a group of Dendrobates tinctorius, a Neotropical anuran species, confiscated from the illegal wildlife trade and housed in a private zoo in Brazil as part of an ex situ breeding program. We examined histological sections of the skin of 30 D. tinctorius and 20 Adelphobates galactonotus individuals. Twenty D. tinctorius (66.7%) and none of the A. galactonotus were positive for Batrachochytrium dendrobatidis (Bd). Multiple development stages of Bd infection were observed. The reasons for the interspecific difference in the rate of infection could not be determined, and further studies are advised. Because the examined population consisted of confiscated frogs, detailed epidemiological aspects could not be investigated, and the source of the fungus remains uncertain. The existence of ex situ amphibian populations is important for protecting species at higher risk in the wild, and ex situ amphibian conservation and breeding programs in Brazil may be established using confiscated frogs as founders. However, this paper alerts these programs to the urgency of strict quarantine procedures to prevent the introduction of potential pathogens, particularly Bd, into ex situ conservation programs.

Sexual dimorphism and directional sexual selection on aposematic signals in a poison frog

It is commonly assumed that natural selection imposed by predators is the prevailing force driving the evolution of aposematic traits. Here, we demonstrate that aposematic signals are shaped by sexual selection as well. We evaluated sexual selection for coloration brightness in populations of the poison frog Oophaga [Dendrobates] pumilio in Panama's Bocas del Toro archipelago. We assessed female preferences for brighter males by manipulating the perceived brightness of spectrally matched males in two-way choice experiments. We found strong female preferences for bright males in two island populations and weaker or ambiguous preferences in females from mainland populations. Spectral reflectance measurements, coupled with an O. pumilio-specific visual processing model, showed that O. pumilio coloration was significantly brighter in island than in mainland morphs. In one of the island populations (Isla Solarte), males were significantly more brightly colored than females. Taken together, these results provide evidence for directional sexual selection on aposematic coloration and document sexual dimorphism in vertebrate warning coloration. Although aposematic signals have long been upheld as exemplars of natural selection, our results show that sexual selection should not be ignored in studies of aposematic evolution.