Development Mode Research Articles

Deuterostomic Development in the Protostome Priapulus caudatus

The fate of the blastopore during development in the bilaterian ancestor is currently not well understood. In deuterostomes, the blastopore forms the anus, but its fate in protostome groups is variable. This variability, combined with an absence of information from key taxa, hampers the reconstruction of the ancestral developmental mode of the Protostomia and the Bilateria. The blastopore fate of the bilaterian ancestor plays a crucial role in understanding the transition from radial to bilateral symmetric organisms. Priapulids have a conservative morphology, an abundant Cambrian fossil record, and a phylogenetic position that make them a key group in understanding protostome evolution. Here, we characterize gastrulation and the embryonic expression of genes involved in bilaterian foregut and hindgut patterning in Priapulus caudatus. We show that the blastopore gives rise to the anus at the vegetal pole and that the hindgut markers brachyury and caudal are expressed in the blastopore and anus, whereas the foregut markers foxA and goosecoid are expressed in the mouth in the animal hemisphere. Thereby, gastrulation in the conservatively evolving protostome P. caudatus follows strictly a deuterostomic pattern. These results are more compatible with a deuterostomic rather than protostomic (blastopore forms the mouth) or amphistomic (mouth and anus are formed simultaneously) mode of development in the last common bilaterian ancestor.

Geographic comparison of seasonal migration events of the large white butterfly, Pieris brassicae

1. The multivoltine butterfly Pieris brassicae (Linnaeus) inhabits most climatic zones in Europe and Asia. Its regional populations are adapted to the respective climatic conditions and hibernate in those regions. Yet how these adaptations are stabilised in spite of the species' regular migration events is unclear.2. In this study, significant differences were found in the preferred flight direction of P. brassicae depending on the season and the butterfly's geographic origin. The individual flight behaviours indicated that flight direction is a heritable character.3. The bivoltine population from northern Germany undertook a typical return flight in the second generation of the year. This reverse flight was induced by the developmental mode of the caterpillars. A diapause in the pupal stage triggered a northward migration, and a non‐diapause a southward migration.4. In contrast to bivoltine populations, the second generation of multivoltine populations maintained the direction of the hibernating generation.5. A unique situation was determined for the population from the northern coast of Spain. The North Sea seemed to be a migration barrier that forced an adaptation, with the first generation flying southeastward, and the second generation northeastward.6. The flight pattern of at least some of the investigated populations suggests that, after their yearly migration, the descendants of a local population are able to reach the home range of their ancestors again.

Segment polarity gene expression in a myriapod reveals conserved and diverged aspects of early head patterning in arthropods

Arthropods show two kinds of developmental mode. In the so-called long germ developmental mode (as exemplified by the fly Drosophila), all segments are formed almost simultaneously from a preexisting field of cells. In contrast, in the so-called short germ developmental mode (as exemplified by the vast majority of arthropods), only the anterior segments are patterned similarly as in Drosophila, and posterior segments are added in a single or double segmental periodicity from a posterior segment addition zone (SAZ). The addition of segments from the SAZ is controlled by dynamic waves of gene activity. Recent studies on a spider have revealed that a similar dynamic process, involving expression of the segment polarity gene (SPG) hedgehog (hh), is involved in the formation of the anterior head segments. The present study shows that in the myriapod Glomeris marginata the early expression of hh is also in a broad anterior domain, but this domain corresponds only to the ocular and antennal segment. It does not, like in spiders, represent expression in the posterior adjacent segment. In contrast, the anterior hh pattern is conserved in Glomeris and insects. All investigated myriapod SPGs and associated factors are expressed with delay in the premandibular (tritocerebral) segment. This delay is exclusively found in insects and myriapods, but not in chelicerates, crustaceans and onychophorans. Therefore, it may represent a synapomorphy uniting insects and myriapods (Atelocerata hypothesis), contradicting the leading opinion that suggests a sister relationship of crustaceans and insects (Pancrustacea hypothesis). In Glomeris embryos, the SPG engrailed is first expressed in the mandibular segment. This feature is conserved in representatives of all arthropod classes suggesting that the mandibular segment may have a special function in anterior patterning.

Development and evolution of the vertebrate primary mouth.

The vertebrate oral region represents a key interface between outer and inner environments, and its structural and functional design is among the limiting factors for survival of its owners. Both formation of the respective oral opening (primary mouth) and establishment of the food-processing apparatus (secondary mouth) require interplay between several embryonic tissues and complex embryonic rearrangements. Although many aspects of the secondary mouth formation, including development of the jaws, teeth or taste buds, are known in considerable detail, general knowledge about primary mouth formation is regrettably low. In this paper, primary mouth formation is reviewed from a comparative point of view in order to reveal its underestimated morphogenetic diversity among, and also within, particular vertebrate clades. In general, three main developmental modes were identified. The most common is characterized by primary mouth formation via a deeply invaginated ectodermal stomodeum and subsequent rupture of the bilaminar oral membrane. However, in salamander, lungfish and also in some frog species, the mouth develops alternatively via stomodeal collar formation contributed both by the ecto- and endoderm. In ray-finned fishes, on the other hand, the mouth forms via an ectoderm wedge and later horizontal detachment of the initially compressed oral epithelia with probably a mixed germ-layer derivation. A very intriguing situation can be seen in agnathan fishes: whereas lampreys develop their primary mouth in a manner similar to the most common gnathostome pattern, hagfishes seem to undergo a unique oropharyngeal morphogenesis when compared with other vertebrates. In discussing the early formative embryonic correlates of primary mouth formation likely to be responsible for evolutionary-developmental modifications of this area, we stress an essential role of four factors: first, positioning and amount of yolk tissue; closely related to, second, endoderm formation during gastrulation, which initiates the process and constrains possible evolutionary changes within this area; third, incipient structure of the stomodeal primordium at the anterior neural plate border, where the ectoderm component of the prospective primary mouth is formed; and fourth, the prime role of Pitx genes for establishment and later morphogenesis of oral region both in vertebrates and non-vertebrate chordates.

Enlightenment of Development of Higher Education in South Korea to China

China and South Korea are two neighboring countries and are both developing countries. Furthermore, the modern and contemporary higher education of these two countries originates from the middle and late 19th Century, so there are a lot of similarities between the two countries. However, although the origination period of modern and contemporary higher education of the two countries is almost similar, the developmental speed of higher education in these two countries is not totally the same due to the different strategic measures taken by the two countries in terms of developmental mode of higher education. Thus, study on differences between the higher education of these two countries and refer to the experience of South Korean higher education is of great and important realistic significance to improve higher education in China and promote and economic and social development. Through an analysis in the development history and educational system of higher education in South Korea, this article makes a comparison of the similarities and differences between China and South Korea in terms of the current condition and reform of their educational system so as to find out the gap between China and South Korea in terms of the higher education and explore a correct path for development of China higher education.

Sipunculan Larvae and "Cosmopolitan" Species

Sipuncula is a relatively small taxon with roughly 150 recognized species. Many species are geographically widespread or "cosmopolitan." The pelagosphera larvae of some species are estimated to spend several months in the plankton. However, recent molecular evidence suggests that many of the "cosmopolitan" species actually represent species-complexes, some not even monophyletic. Herein, we present data on three sipunculan species with different developmental modes that occur both in the Sea of Japan and in the Northeast Pacific. The development of the three species-Phascolosoma agassizii, Thysanocardia nigra, and Themiste pyroides-is exceptionally well studied in both regions of the Pacific. Significant differences have been observed between the two regions with respect to egg size, developmental mode, and developmental timing. In general, eggs are larger and development slower in the Northeast Pacific when compared with the Sea of Japan. These differences have been explained as a result of phenotypic plasticity exhibited under different environmental conditions, in particular temperature, but we show that the populations of all three species are also remarkably distinct genetically and that gene flow between the two regions is extremely unlikely. In Thysanocardia nigra, we even found two very distinct genetic lineages within the same location in the Northeast Pacific. The amount of genetic divergence between populations from the Sea of Japan and those from the Northeast Pacific is not correlated with developmental mode. Themiste pyroides, the species with the most abbreviated development, actually has the least degree of genetic divergence between the regions. Analyses of molecular variance show that the majority of the observed variation in all three species is between the regions. We conclude that all three "cosmopolitan" species actually represent complexes of cryptic or pseudo-cryptic species. These examples demonstrate that a solid taxonomic framework based on molecular and morphological evidence is a prerequisite for evaluating relationships between dispersal capabilities, species' ranges, and the connectivity of populations.

Polymorphism in Developmental Mode and Its Effect on Population Genetic Structure of a Spionid Polychaete, Pygospio elegans

Population genetic structure of sedentary marine species is expected to be shaped mainly by the dispersal ability of their larvae. Long-lived planktonic larvae can connect populations through migration and gene flow, whereas species with nondispersive benthic or direct-developing larvae are expected to have genetically differentiated populations. Poecilogonous species producing different larval types are ideal when studying the effect of developmental mode on population genetic structure and connectivity. In the spionid polychaete Pygospio elegans, different larval types have been observed between, and sometimes also within, populations. We used microsatellite markers to study population structure of European P. elegans from the Baltic Sea (BS) and North Sea (NS). We found that populations with planktonic larvae had higher genetic diversity than did populations with benthic larvae. However, this pattern may not be related to developmental mode, since in P. elegans, developmental mode may be associated with geography. Benthic larvae were more commonly seen in the brackish BS and planktonic larvae were predominant in the NS, although both larval types also are found from both areas. Significant isolation-by-distance (IBD) was found overall and within regions. Most of the pair-wise F(ST) comparisons among populations were significant, although some geographically close populations with planktonic larvae were found to be genetically similar. However, these results, together with the pattern of IBD, autocorrelation within populations, as well as high estimated local recruitment, suggest that dispersal is limited in populations with planktonic larvae as well as in those with benthic larvae. The decrease in salinity between the NS and BS causes a barrier to gene flow in many marine species. In P. elegans, low, but significant, differentiation was detected between the NS and BS (3.34% in AMOVA), but no clear transition zone was observed, indicating that larvae are not hampered by the change in salinity.

Sustainable development and upgrading mode of coal industry in China

It is an emerging realistic problem on how to promote a high level of technology in the coal industry, find new upgrading powers and create new competitive advantages, which are also the core problems to efficient transformation pattern of economic growth for coal industry in the “twelfth five-year” period, involving the key to China’s energy supply and energy security. Through field surveys and inductive analyses, analyzing of the content of upgrading the coal industry as an entry point, this paper analyzes the sustainable development mechanisms for the coal industry from the aspects of work force, power, methods, goals, and so on; and puts forward corresponding developmental modes according to the mechanisms which can improve resource recovery and mineral resources utilization rates by putting them into practice.

Study on the Strategies of the Spatial Development Mode of the Creative Park Derived from the Creative Industries

With the booming development of creative industries, the creative park - the spatial form of this new era-plays a fairly important role in modern urban life. Creative park not only performs the responsibilities of constantly making deep dialogue and communication between people and city’s cultural life, but also makes a variety of types of space achieve regenerative transformation and functional transformation. After the early stages of development, the present problems the creative park face is how to survive with an attitude of sustainable development and a positive spatial development modes. This article aims to summarize some constructive developmental mode strategies on the basis of finished cases, which has certain positive and practical significance for the creative park space derived from the creative industries.

Histamine is a modulator of metamorphic competence in Strongylocentrotus purpuratus (Echinodermata: Echinoidea).

BackgroundA metamorphic life-history is present in the majority of animal phyla. This developmental mode is particularly prominent among marine invertebrates with a bentho-planktonic life cycle, where a pelagic larval form transforms into a benthic adult. Metamorphic competence (the stage at which a larva is capable to undergo the metamorphic transformation and settlement) is an important adaptation both ecologically and physiologically. The competence period maintains the larval state until suitable settlement sites are encountered, at which point the larvae settle in response to settlement cues. The mechanistic basis for metamorphosis (the morphogenetic transition from a larva to a juvenile including settlement), i.e. the molecular and cellular processes underlying metamorphosis in marine invertebrate species, is poorly understood. Histamine (HA), a neurotransmitter used for various physiological and developmental functions among animals, has a critical role in sea urchin fertilization and in the induction of metamorphosis. Here we test the premise that HA functions as a developmental modulator of metamorphic competence in the sea urchin Strongylocentrotus purpuratus.ResultsOur results provide strong evidence that HA leads to the acquisition of metamorphic competence in S. purpuratus larvae. Pharmacological analysis of several HA receptor antagonists and an inhibitor of HA synthesis indicates a function of HA in metamorphic competence as well as programmed cell death (PCD) during arm retraction. Furthermore we identified an extensive network of histaminergic neurons in pre-metamorphic and metamorphically competent larvae. Analysis of this network throughout larval development indicates that the maturation of specific neuronal clusters correlates with the acquisition of metamorphic competence. Moreover, histamine receptor antagonist treatment leads to the induction of caspase mediated apoptosis in competent larvae.ConclusionsWe conclude that HA is a modulator of metamorphic competence in S. purpuratus development and hypothesize that HA may have played an important role in the evolution of settlement strategies in echinoids. Our findings provide novel insights into the evolution of HA signalling and its function in one of the most important and widespread life history transitions in the animal kingdom - metamorphosis.

Five questions about Streptococcus mutans: Theoretical study of its transmission and colonisation

PurposeThis theoretical study aimed to identify the decisive (and controllable) factors involved in Streptococcus mutans (Sm) infection through addressing questions about (i) the time and prevalence pattern (including the raison d’etre of the discrete period for the infection or WI) of initial Sm colonisation and (ii) the infant's selection of bacterial types and their diversity, which are not yet definitely answered by empirical works. MethodA model of Sm infection (within-host type) was developed. For questions (i): using the basic model, stochastic simulation was performed to reproduce longitudinal observations of the initial colonisation time. A symmetrical or right-skewed gamma distribution was assumed for the maximum colonisable area (Kmax) and transmission rate (mx). Additionally, 3 or 4 developmental modes of colonisable area [K(t)] were assigned based on the Kmax value. For (ii): by extending the basic model to the two-bacterial type model, intraspecific competition analysis focusing on the differences in mx (received by the infancy) and colonisation ability (θD) was performed. ResultsThe basic model simulation showed that mx and K(t) played a pivotal role in determining the individual time of initial colonisation and their variations among infants in forming its prevalence patterns (with or without WI). The competition model simulation showed that higher mx could be more advantageous in competitive colonisation than higher θD under repeated invasions. Accordingly, it played a decisive role in infant's selection of initially, persistently and transiently colonising bacterial types, and thus in their diversity. Conclusions(i) The mx is the primary and controllable (risk) factor that extensively affects various aspects of the Sm infection process. (ii) Also, the growing carrying capacity, i.e., K(t) is another important factor when considering how to effectively delay the onset of the colonisation. (iii) Thus, currently, the most feasible and effective control measure for the infection should be microbiological interventions in the primary host with concurrent oral hygiene and dietary control in the exposed child.

The situation and consideration about pre-hospital emergency transfer of children

It is a brief introduction of different transfer modes with focus on ranges,measures,quality control,systems,training and process of transfer system of children according to the situation in China even abroad.With analysis of main problems and causes of children' transfer system,the discussion contained management of pre-hospital transfer system,risk assessment of critical patients,establishment of transfer network of critically ill children,the relationship of first aid and pre-hospital transfer,an effective response to public health emergencies of children as well as developmental mode for emergency transfer of children. Key words: Pre-hospital emergency transfer; Children

Phylogenetic analysis of cryptic speciation in the polychaete Pygospio elegans

Development in marine invertebrate species can take place through a variety of modes and larval forms, but within a species, developmental mode is typically uniform. Poecilogony refers to the presence of more than one mode of development within a single species. True poecilogony is rare, however, and in some cases, apparent poecilogony is actually the result of variation in development mode among recently diverged cryptic species. We used a phylogenetic approach to examine whether poecilogony in the marine polychaete worm, Pygospio elegans, is the result of cryptic speciation. Populations of worms identified as P. elegansooded, and intermediate larvae; these modes are found both within and among populations. We examined sequence variation among partial mitochondrial cytochrome c oxidase subunit I sequences obtained for 279 individual worms sampled across broad geographic and environmental scales. Despite a large number of unique haplotypes (121 haplotypes from 279 individuals), sequence divergence among European samples was low (1.7%) with most of the sequence variation observed within populations, relative to the variation among regions. More importantly, we observed common haplotypes that were widespread among the populations we sampled, and the two most common haplotypes were shared between populations differing in developmental mode. Thus, our results support an earlier conclusion of poecilogony in P elegans. In addition, predominantly planktonic populations had a larger number of population-specific low-frequency haplotypes. This finding is largely consistent with interspecies comparisons showing high diversity for species with planktonic developmental modes in contrast to low diversity in species with brooded developmental modes.

Introduction to Symposium: Poecilogony--A Window on Larval Evolutionary Transitions in Marine Invertebrates

Poecilogony is the intraspecific variation in developmental mode that has been described in some marine invertebrates. Poecilogonous species produce different larval forms (e.g., free-swimming planktotrophic larvae as well as brooded lecithotrophic or adelphophagic larvae). Poecilogony can be a controversial topic, since it is difficult to identify and characterize the phenomenon with certainty. It has been challenging to determine whether poecilogony represents developmental polymorphism with a genetic basis or developmental polyphenism reflecting plastic responses to environmental cues. Other outstanding questions include whether common mechanisms underlie the developmental variation we observe in poecilogonous species, and whether poecilogony is maintained in different taxa through similar mechanisms or selective pressures. Poecilogonous species provide a unique opportunity to elucidate the cellular, developmental, and genetic mechanisms underlying evolutionary transitions in developmental mode, as well as to help clarify the selective pressures and possible ecological circumstances that might be involved. Here, we describe an integrative approach to the study of poecilogony and its role in larval evolutionary transitions highlighted during a symposium held at the 2012 annual meeting of the Society for Integrative and Comparative Biology.

Marine Mollusca of the last 2 million years in New Zealand. Part 5. Summary

Evolution occurred equally through allopatric speciation and anagenesis during Pliocene–Early Pleistocene time. However, no anagenesis is observed in Castlecliffian to Recent Mollusca. Possible explanations include (1) increasing shallowness and energy of the environment (Sheldon's ‘plus ça change’ model); (2) lecithotrophs increased with falling temperatures, and are more susceptible than planktotrophs to allopatric speciation; (3) ‘plus ça change’ implies that anagenesis continued unabated, but unrecorded, offshore from the recorded Pleistocene fauna. Three lineages (Pelicaria vermis, Stiracolpus symmetricus, Aoteadrillia wanganuiensis) became highly variable as a result of their changed developmental mode. Origination continued throughout Castlecliffian–Haweran time at a background rate of 0–2 new species per oxygen isotope stage (OIS), whereas extinction and range extensions of warm-water taxa to Wanganui Basin both reached a peak in OIS 9–7. However, the faunal record results from the interplay of temperature change and the preservation of different depositional environments. Biostratigraphy and temperature change also are reviewed.

Transcriptome Analysis and SNP Development Can Resolve Population Differentiation of Streblospio benedicti, a Developmentally Dimorphic Marine Annelid

Next-generation sequencing technology is now frequently being used to develop genomic tools for non-model organisms, which are generally important for advancing studies of evolutionary ecology. One such species, the marine annelid Streblospio benedicti, is an ideal system to study the evolutionary consequences of larval life history mode because the species displays a rare offspring dimorphism termed poecilogony, where females can produce either many small offspring or a few large ones. To further develop S. benedicti as a model system for studies of life history evolution, we apply 454 sequencing to characterize the transcriptome for embryos, larvae, and juveniles of this species, for which no genomic resources are currently available. Here we performed a de novo alignment of 336,715 reads generated by a quarter GS-FLX (Roche 454) run, which produced 7,222 contigs. We developed a novel approach for evaluating the site frequency spectrum across the transcriptome to identify potential signatures of selection. We also developed 84 novel single nucleotide polymorphism (SNP) markers for this species that are used to distinguish coastal populations of S. benedicti. We validated the SNPs by genotyping individuals of different developmental modes using the BeadXPress Golden Gate assay (Illumina). This allowed us to evaluate markers that may be associated with life-history mode.

A global molecular phylogeny of 147 periwinkle species (Gastropoda, Littorininae)

Reid, D. G., Dyal, P. & Williams, S.T. (2012) A global molecular phylogeny of 147 periwinkle species (Gastropoda, Littorininae). —Zoologica Scripta, 41, 125–136.Complete species‐level molecular phylogenies have been published for several genera of Littorinidae (e.g. Echinolittorina, Littoraria). Here we add new sequence data from three genes (28S rRNA, 12S rRNA, cytochrome oxidase c subunit I) for single specimens of an additional 24 species, to make a data set of 147 (97%) of the 152 recognized species of the subfamily Littorininae. This three‐gene data set is analysed to produce a phylogenetic hypothesis for the subfamily, which includes the first complete species‐level phylogeny of the genus Peasiella and the first three‐gene phylogeny of all Littorina species. The non‐planktotrophic species of Littorina have previously been classified together (as subgenus Neritrema), implying a single origin of this developmental mode. Tests of this hypothesis with the new data are inconclusive, and resolution is not improved in a tree constructed from five genes (adding previously published sequences of 16S rRNA and cytochrome b). Using available fossils for calibration we generate a BEAST chronogram, which emphasizes that the radiation of Littorina is more recent than that of other littorinine genera. A database is provided, listing all known species of Littorininae, with their distributions, development, ecology and gene sequences, as a tool for future evolutionary studies of this model group.

Egg Size, Life History, and Tropical Marine Gastropod Biogeography*

Comparative studies of egg size and larval morphological characteristics coupled with estimates of dispersal abilities enabled Kohn and Perron (1994) to predict attributes of geographic distribution patterns of Indo-West Pacific Conus species that vary widely in developmental mode. However, because at that time, no species-level phylogenetic hypothesis had ever been proposed for Conus, we were constrained to treat each species as an independent entity. During the past decade, however, molecular phylogenetics of Conus has progressed to the point where character mapping and the method of phylogenetically independent contrasts can evaluate the importance of phylogenetic relatedness to biogeographic patterns. Analyses of species in two well-supported clades with data on ranges of trait variations suggest that significant evolutionary associations remain among developmental traits that affect dispersal ability after accounting for phylogenetic relationships. These are clades of molluscivorous and piscivorous Conus species, both exclusively in the Indo-Pacific region. As yet inadequate data on both development types and biogeography exist to determine whether phylogenetic relatedness predicts biogeographic patterns elsewhere in this hyperdiverse genus.

Expression of pair rule gene orthologs in the blastoderm of a myriapod: evidence for pair rule-like mechanisms?

BackgroundA hallmark of Drosophila segmentation is the stepwise subdivision of the body into smaller and smaller units, and finally into the segments. This is achieved by the function of the well-understood segmentation gene cascade. The first molecular sign of a segmented body appears with the action of the pair rule genes, which are expressed as transversal stripes in alternating segments. Drosophila development, however, is derived, and in most other arthropods only the anterior body is patterned (almost) simultaneously from a pre-existing field of cells; posterior segments are added sequentially from a posterior segment addition zone. A long-standing question is to what extent segmentation mechanisms known from Drosophila may be conserved in short-germ arthropods. Despite the derived developmental modes, it appears more likely that conserved mechanisms can be found in anterior patterning.ResultsExpression analysis of pair rule gene orthologs in the blastoderm of the pill millipede Glomeris marginata (Myriapoda: Diplopoda) suggests that these genes are generally involved in segmenting the anterior embryo. We find that the Glomeris pairberry-1 ( pby-1) gene is expressed in a pair rule pattern that is also found in insects and a chelicerate, the mite Tetraynchus urticae. Other Glomeris pair rule gene orthologs are expressed in double segment wide domains in the blastoderm, which at subsequent stages split into two stripes in adjacent segments.ConclusionsThe expression patterns of the millipede pair rule gene orthologs resemble pair rule patterning in Drosophila and other insects, and thus represent evidence for the presence of an ancestral pair rule-like mechanism in myriapods. We discuss the possibilities that blastoderm patterning may be conserved in long-germ and short-germ arthropods, and that a posterior double segmental mechanism may be present in short-germ arthropods.

Nidicolous tadpoles rather than direct development in Malagasy frogs of the genus Gephyromantis

Frogs in the genus Gephyromantis from Madagascar were assumed to have a direct developmental mode, i.e. the complete embryonic and larval development within the egg, but recently free-swimming, exotrophic tadpoles of a few species have been found. Herein we provide detailed morphological descriptions of the tadpoles of five more species of this genus, indicating a developmental mode other than direct development in species of Gephyromantis. Tadpoles of Gephyromantis granulatus, G. sculpturatus, G. tschenki, and G. ventrimaculatus were found free-swimming in streams, and tadpoles of G. sp. aff. blanci were raised after hatching from clutches found in the leaf litter. All tadpoles were identified by DNA barcoding. The oral discs of all five species are characterized by the lack of many typical morphological traits of exotrophic tadpoles (such as oral papillae and keratodonts). This indicates that these tadpoles are either non-feeding (endotrophic) or only facultatively feeding tadpoles. We classify these tadpoles as nidicolous based on the observation that the larvae of G. sp. aff. blanci stayed after hatching in the jelly nest until metamorphosis. It remains unclarified whether all species have strictly nidicolous tadpoles, and the larvae of the four species found in the streams were just accidentally washed into these streams; alternatively, some of these tadpoles might be nidicolous at first but in some species need to live in free water at later stages.