DNA barcode reveals candidate species of Scinax and Ololygon (Anura: Hylidae) in Atlantic Forest

Molecular species delimitation methods are efficient tools to identify species, including the discovery of new taxa and cryptic organisms, thus being useful to biodiversity studies. In the present work, 16S mitochondrial sequences and cytochrome oxidase I (COI) were used to evaluate the richness of species in the genus Scinax and Ololygon from a biodiversity hotspot in Atlantic Forest. A total of 109 specimens formally belonging to eight species of Scinax and three species of Ololygon were collected in 13 localities along the state of Bahia (northeastern Brazil) and one site in Espírito Santo (southeastern Brazil). Of the Scinax species collected in this study, three were morphologically differentiated from other described species and identified as putative new species (Scinax sp.1, Scinax sp.2 and Scinax sp.3). The species delimitations were inferred using three different methods: ABGD, PTP and mPTP which allowed recognizing 11 Scinax species and five Ololygon species. Scinax sp. 1, Scinax sp. 2 and Scinax sp. 3, have been confirmed as new putative species and Ololygon argyreornata possibly contains cryptic species. We suggest additional studies, including morphological and bioacoustic data to validate these new putative species.     

An historical interpretation of habitat use by frogs in a Central Amazonian Forest

Interpretations of habitat use in tropical frog assemblages have centred on resource partitioning and stressed the influence of interspecific interaction and climatic fluctuation on numbers of species using various habitats. We used audio strip transects and visual methods to determine the species composition, reproductive modes, and habitat occupancy patterns of the entire assemblage of frog species in 1900 hectares of primary forest north of Manaus in the central Amazon. We then compared taxon, reproductive mode, and habitat of species at six analogous lowland forest sites of similar species richness (five in the Amazon and one in Southeast Asia) to determine similarity of habitat use among sites and whether habitat is strongly associated with species» systematic positions. In all lowland Amazonian faunas, most species with aquatic development use pools, many species undergo some degree of terrestrial development, and few species are riparian or develop in streams. In contrast, about half the species in Southeast Asian assemblages are riparian and develop in streams, and few species develop terrestrially. Because reproductive mode and habitat associate strongly with taxon, patterns of habitat use observed at this regional scale are better explained by historical biogeography and differential rates of speciation than by proximal selection generated by contemporary environmental conditions. This study presents an inventory of frog species in a central Amazonian terre-firme forest and measurements of habitat availability and use by an entire assemblage of frogs throughout a large area (other portions of this study were published by Gascon, 1990, 1991; Zimmerman & Rodrigues, 1990; Zimmerman, 1991). We asked whether this local pattern of habitat occupancy differed from the regional Amazonian pattern and whether local species composition could be predicted from (sub)habitat composition. Viewing the assemblage at the local level did reveal species-(sub)habitat relationships masked at the broader regional level. About half the pool-breeders at the Manaus forest study sites would not use pools that could be flooded by a permanent stream; several species distinguished between permanent and temporary ponds; and some species occupied all available breeding habitat, whereas others occurred patchily. This pattern was maintained over four breeding seasons, and species composition could be predicted from (sub)habitat composition. Phylogeny was not a predictor of subhabitat occupancy. Perhaps species are phylogenetically constrained to develop in pool, stream, riparian, or terrestrial habitats, but contemporary selection governs their narrow distribution within these major habitat types. Finally, we asked whether anuran species richness in the central Amazon differs from that of the upper or lower Amazon. One genus, Eleutherodactylus , accounts for elevated species richness at upper Amazonian sites. Dry seasons in the central and lower Amazon are unlikely to restrict the spread of eleutherodactylines, which reproduce terrestrially. There are as many non-eleutherodactylines with terrestrial development at seasonal sites as there are at continually wet sites. Colonization history and the topography of central and lower Amazonia are more likely to limit eleutherodactyline richness.     

Phylogenetic origins of local-scale diversity patterns and the causes of Amazonian megadiversity

What explains the striking variation in local species richness across the globe and the remarkable diversity of rainforest sites in Amazonia? Here, we apply a novel phylogenetic approach to these questions, using treefrogs (Hylidae) as a model system. Hylids show dramatic variation in local richness globally and incredible local diversity in Amazonia. We find that variation in local richness is not explained primarily by climatic factors, rates of diversification (speciation and extinction) nor morphological variation. Instead, local richness patterns are explained predominantly by the timing of colonization of each region, and Amazonian megadiversity is linked to the long-term sympatry of multiple clades in that region. Our results also suggest intriguing interactions between clade diversification, trait evolution and the accumulation of local richness. Specifically, sympatry between clades seems to slow diversification and trait evolution, but prevents neither the accumulation of local richness over time nor the co-occurrence of similar species.     

Discovering Hidden Diversity of Characins (Teleostei: Characiformes) in Ecuador’s Yasuní National Park

Background

Management and conservation of biodiversity requires adequate species inventories. The Yasuní National Park is one of the most diverse regions on Earth and recent studies of terrestrial vertebrates, based on genetic evidence, have shown high levels of cryptic and undescribed diversity. Few genetic studies have been carried out in freshwater fishes from western Amazonia. Thus, in contrast with terrestrial vertebrates, their content of cryptic diversity remains unknown. In this study, we carried out genetic and morphological analyses on characin fishes at Yasuní National Park, in eastern Ecuador. Our goal was to identify cryptic diversity among one of the most speciose fish families in the Amazon region. This is the first time that genetic evidence has been used to assess the species content of the Napo Basin, one of the richest regions in vertebrate diversity.

Results

Phylogenetic analyses of partial mitochondrial 16S ribosomal RNA gene (∼600 pb) DNA sequences from 232 specimens of the family Characidae and its closest groups revealed eight candidate new species among 33 species sampled, representing a 24% increase in species number. Analyses of external morphology allowed us to confirm the species status of six of the candidate species.

Conclusions

Our results show high levels of cryptic diversity in Amazonian characins. If this group is representative of other Amazonian fish, our results would imply that the species richness of the Amazonian ichthyofauna is highly underestimated. Molecular methods are a necessary tool to obtain more realistic inventories of Neotropical freshwater fishes.     

Amphibian diversity in Polish cities: Taxonomic diversity,functional diversity and evolutionary distinctiveness

As a process affecting animal communities, urbanization has been the subject of numerous studies. However, amphibians are still among the least studied vertebrate groups in urbanized landscapes. Generally, it has been found that the process of loss of amphibian diversity is nonrandom, with species from older evolutionary lines at greater risk. Regional data on amphibian assemblages in urban areas is a very useful tool for assessing how these assemblages react to changes.The aim of the present paper is to assess the diversity of amphibians in Polish cities based on data in the relevant literature, exploring different metrics (e.g., taxonomic, functional, and evolutionary diversity) calculated in amphibian species assemblages. We used data from 18 articles (including grey literature), characterized by comparable research methods and published between 1999 and 2017.Overall, amphibian species richness (ASR) amounted to an average of 9 species, ranging from 5 to 11 species per city. The higher species richness occurred in Białystok in North-west of Poland. Functional evenness (FEve), evolutionary distinctiveness (ED sum), and functional richness (FRic) were strongly positively correlated with ASR. However, ED mean was not significantly correlated with the total number of species in the community. Three taxa, the hybridogenetic water frogs Pelophylax esculentus complex, the common toad Bufo bufo, and the common frog Rana temporaria, occurred in all analyzed amphibian assemblages. Our study is one of the first attempts to compare urban amphibian assemblages, using different and complementary diversity metrics on a large spatial scale. In conclusion, we highlight that urban areas play an important role for conservation of amphibians, because they support amphibian assemblages characterized by a high level of overall diversity.     

The importance of considering both taxonomic and habitat guild approaches in small mammal research

Studies on the effects of habitat fragmentation on small mammals often lead to confounding results as they only consider taxonomic groups in their analysis and neglect functional diversity of the communities. Here we describe the structure and composition of small mammal communities at 22 sites, ranging from 41 to 7035 ha, in a hyper‐fragmented landscape of an Amazonia‐Cerrado ecotone. Also, in considering a taxonomic and habitat guild approach, we report the effects of habitat structures and patch spatial attributes on richness, abundance and species composition. Small mammal richness reported in southern Amazonia (N = 23 species) is greater than most previous studies in the tropics. All rare small mammals captured in this study were forest interior species. Richness of forest interior species was positively related to larger patches, as shown by the species–area relationship. However, 52% of the small mammal species were in forest fragments smaller than 50 ha, highlighting the importance of preserving both large and small forest fragments in a landscape with accelerated habitat reduction. Richness of edge‐tolerant species was not associated with the tested variables, yet edge‐tolerant species were more abundant in degraded environments. Marsupials were positively associated with vertical habitat structures, while rodents were more strongly related to a ground‐level habitat structure. The landscape studied is extremely variable and has contributed to the difficulty in detecting clear patterns, particularly when considering only one approach. Because of the complementary outputs when analysing either taxonomic groups or habitat guilds, we recommend the use of multi‐taxa studies of different guilds to assist decision makers in designing conservation strategies and appropriate management of small mammal populations.     

Locality or habitat? Exploring predictors of biodiversity in Amazonia

Amazonia is an environmentally heterogeneous and biologically megadiverse region, and its biodiversity varies considerably over space. However, existing knowledge on Amazonian biodiversity and its environmental determinants stems almost exclusively from studies of macroscopic above‐ground organisms, notably vertebrates and trees. In contrast, diversity patterns of most other organisms remain elusive, although some of them, for instance microorganisms, constitute the overwhelming majority of taxa in any given location, both in terms of diversity and abundance. Here, we use DNA metabarcoding to estimate prokaryote and eukaryote diversity in environmental soil and litter samples from 39 survey plots in a longitudinal transect across Brazilian Amazonia using 16S and 18S gene sequences, respectively. We characterize richness and community composition based on operational taxonomic units (OTUs) and test their correlation with longitude and habitat. We find that prokaryote and eukaryote OTU richness and community composition differ significantly among localities and habitats, and that prokaryotes are more strongly structured by locality and habitat type than eukaryotes. Our results 1) provide a first large‐scale mapping of Amazonian soil biodiversity, suggesting that OTU richness patterns might follow substantially different patterns from those observed for macro‐organisms; and 2) indicate that locality and habitat factors interact in determining OTU richness patterns and community composition. This study shows the potential of DNA metabarcoding in unveiling Amazonia's outstanding diversity, despite the lack of complete reference sequence databases for the organisms sequenced.     

Molecular systematics and phylogeography of Amazonian poison frogs of the genus Dendrobates

The study of Amazonian biodiversity requires detailed knowledge of the phylogenetic relationships of closely related taxa distributed across Amazonia. The Amazonian poison frogs of the genus Dendrobates have undergone many taxonomic revisions, but the phylogenetic relationships within this group remain poorly understood. Most previous classifications were based on morphology and skin toxin analyses, with limited use of DNA sequence data. Using mtDNA sequence data from four gene regions (cytochrome b, cytochrome oxidase I, 16S rRNA, and 12S rRNA), we present a molecular phylogenetic analysis of the evolutionary relationships within a representative group of Amazonian Dendrobates. We use the resulting phylogenetic hypothesis to investigate different biogeographic hypotheses concerning genetic divergence and species diversity in Amazonia. The results of the analysis support the presence of ancient paleogeographic barriers to gene flow between eastern and western Amazonia, and indicate substantial genetic divergence between species found in the northern and southern regions of western Amazonia.     

Useful Species Richness,Proportion of Exotic Species,and Market Orientation on Amazonian Dark Earths and Oxisols<Superscript>1</Superscript>

Useful Species Richness, Proportion of Exotic Species, and Market Orientation on Amazonian Dark Earths and Oxisols Anthropogenic soils of Amazonia, known as Amazonian Dark Earths (ADE), are environments with elevated soil fertility that can produce crops that otherwise yield poorly on the leached and highly acidic Oxisols that dominate much of the basin. While ADE sites near urban centers often attract commercial horticultural production of nutrient–demanding exotics, these soils are also considered possibly unique reservoirs of endemic agrobiodiversity because of their relationship to pre–Columbian indigenous occupation. Through botanical surveys and interviews with smallholder farmers, this study compared useful species richness, proportion of exotic species, and market orientation of farms situated on ADE and non–anthropogenic Oxisols in the municipality of Borba in Central Brazilian Amazonia. Species richness was similar on Amazonian Dark Earth and Oxisol farms (19.6 spp vs. 18.3 spp); however, ADE farms showed significantly higher proportions of exotic species (39% vs. 26%; p = 0.025). Furthermore, ADE farms in Borba demonstrated significantly higher market orientation (61.0% vs. 47.3%; p = 0.028), likely a result of the advantage of Amazonian Dark Earths for production of crops that are nutrient–demanding or pH–sensitive crops that have higher values in the nearby regional market of Manaus.     

Reproductive habitat selection in alien and native populations of the genus Discoglossus

The existence of suitable breeding habitats is an important factor explaining the regional presence of an anuran species. This study examined patterns of habitat selection in populations of three species of the genus Discoglossus: Discoglossus galganoi (south-western Iberian Peninsula), Discoglossus scovazzi (Morocco) and Discoglossus pictus (three different areas were included in the study: Sicily, Tunisia and north-eastern Iberian Peninsula). The populations of D. pictus on the Iberian Peninsula are allochthonous, and analysis of these patterns may provide insights into the processes that regulate the invasion phase. The hypotheses tested were: (i) congeneric species show the same patterns of habitat selection, and alien species have been established following these patterns; (ii) there are differences in species associations between assemblages structured deterministically and by chance, i.e. native versus invaded assemblages. The larval habitats of three species of this genus were characterized by measuring physical and chemical parameters of the water bodies. We examined the covariation between the presence of Discoglossus species and the species richness of sympatric anurans, and investigated a possible relationship between morphological similarity (as a proxy of functional group) and overlap in habitat use. The results showed that congeneric species are morphologically conservative and also select very similar types of aquatic habitat. The alien population and other sympatric species showed a high degree of overlap in habitat use, which was greater than that observed in the native assemblage with a similar functional richness. Species associations were not structured on the basis of morphological similarity in any of the assemblages. Among native populations, the presence of Discoglossus was either negatively correlated or not significantly correlated with species richness. Only the alien population showed a positive correlation between its presence and species richness, which suggests a loss of assemblage structure.     

Patterns of speciation are similar across mountainous and lowland regions for a Neotropical plant radiation (Costaceae: Costus)

High species richness and endemism in tropical mountains are recognized as major contributors to the latitudinal diversity gradient. The processes underlying mountain speciation, however, are largely untested. The prevalence of steep ecogeographic gradients and the geographic isolation of populations by topographic features are predicted to promote speciation in mountains. We evaluate these processes in a species-rich Neotropical genus of understory herbs that range from the lowlands to montane forests and have higher species richness in topographically complex regions. We ask whether climatic niche divergence, geographic isolation, and pollination shifts differ between mountain-influenced and lowland Amazonian sister pairs inferred from a 756-gene phylogeny. Neotropical Costus ancestors diverged in Central America during a period of mountain formation in the last 3 million years with later colonization of Amazonia. Although climatic divergence, geographic isolation, and pollination shifts are prevalent in general, these factors do not differ between mountain-influenced and Amazonian sister pairs. Despite higher climatic niche and species diversity in the mountains, speciation modes in Costus appear similar across regions. Thus, greater species richness in tropical mountains may reflect differences in colonization history, diversification rates, or the prevalence of rapidly evolving plant life forms, rather than differences in speciation mode.     

Responses of Primates to Landscape Change in Amazonian Land‐bridge islands—a Multi‐scale Analysis

Large hydroelectric dams are one of the current drivers of habitat loss across Amazonian forests. We investigated how the primate community at a hydroelectric dam in Brazilian Amazonia responded to changes in the landscape and local habitat structure of land‐bridge islands after 21 yr of post‐isolation history. The Balbina Dam, constructed in 1986, inundated 3129 km² of primary forests and created more than 3500 variable‐sized islands. We conducted primate and habitat structure surveys on 20 islands from 5 to 1815 ha, and extracted forest patch and landscape metrics for each island. The number of primate species per island varied between 0 and 7 species. Primate composition varied substantially according to both island area and forest cover remaining within the landscape, whereas island area alone was the most significant predictor of richness. Locally, tree density and vertical stratification were the most significant explanatory variables of primate composition and richness. A model containing area effects had the most explanatory power regarding site occupancy for most species. Individually, each species responded differently, with howler and brown capuchin monkeys showing greater tolerance to cope with habitat changes. Body size was also an important predictor of primate occupancy. We recommend protecting large fragments and enhancing the suitability of surrounding habitats to ensure primate conservation in most Neotropical fragmented landscapes. Given the flat topography of hydroelectric reservoirs, which mainly favors the formation of small islands, and the escalating hydropower development plans in Amazonia, our findings provide evidence for pervasive detrimental impacts of dams on primate communities.     

Dacryodes edilsonii, a new species from southwestern Amazonia. Studies in neotropical Burseraceae XII

Dacryodes edilsonii Daly (Burseraceae) is one of at least 14 still undescribed neotropical species in the genus; it is also one of a large number of new species emerging from intensified botanical exploration of the Brazilian state of Acre in southwestern Amazonia. The new species is easily distinguished from most other Neotropical species of the genus by its usually densely long-pubescent petiole, rachis, petiolules, and abaxial midvein.     

Low species turnover of upland Amazonian birds in the absence of physical barriers

Aim

One of the oldest and most powerful ways for ecologists to explain distinct biological communities is to invoke underlying environmental differences. But in hyper-diverse systems, which often display high species richness and low species abundance, these sorts of community comparisons are especially challenging. The classic view for Amazonian birds posits that riverine barriers and habitat specialization determine local and regional community composition. We test the tacit, complementary assumption that similar bird communities should therefore permeate uniform habitat between major rivers, regardless of distance.

Location

Upland (terra firme) rainforests of central Amazonia.

Methods

We conducted intensive whole-community surveys of birds in three pairs of 100-ha plots, separated by 40–60 km. We then used dissimilarity indices, cluster analysis, and ordination to characterize differences among the six avian communities.

Results

In all, we detected 244 forest-dependent birds, with an average of 190 species (78%) per plot. Species turnover was negligible, no unique indicator species were found among plot pairs, and all documented species were already known from a complete inventory at one of the three sites.

Main Conclusions

Our study corroborates the classic biogeographical pattern and suggests that turnover contributes little to regional avian diversity within upland forests. Using a grain size of 100 ha, this implies that upland birds perceive the environment as uniform, at least over distances of ~60 km. Therefore, to maximize both local species richness and population persistence, our findings support the conservation of very large tracts of upland rainforest. Our analyses also revealed that the avifauna at Reserva Ducke, encroached by urban sprawl from the city of Manaus, shows the hallmarks of a disturbed community, with fewer vulnerable insectivores. This defaunation signals that even an enormous preserve (10 × 10 km) in lowland Amazonia is not insulated from anthropogenic degradation within the surrounding landscape.     

The use of a functional approach as surrogate of Collembola species richness in European perennial crops and forests

Collembolans are known indicators of soil disturbance, used in several soil biodiversity monitoring programmes. As for other groups of soil microarthropods, taxonomic determination of Collembola species requires a huge effort and expert knowledge. In this study, we evaluated whether identification of Collembola species to the morphotype level, using an eco-morphological index which classifies individuals based on their adaptation to the soil, can be used as a surrogate of species richness in extensive monitoring schemes. The same evaluation was performed for higher taxa surrogates, using taxonomic categories at the genus and family levels. Additionally, sampling effort for perennial crops and forest systems was determined. Species data were collected from 35 sites sampled within different projects with the same number of samples (16 per site). Results showed that, on average, 8–12 samples are enough to have a good estimate of species richness for this type of systems, averaging 80–90%, but varying considerably with site habitat heterogeneity and local Collembola species pool. GLM models (using species richness as response variable) fitted for family, genus or morphotype levels (explanatory variables) were all significant (p < 0.05) but only the latter two had a pseudo R² higher than 0.75. This indicates that when a rapid, cost-effective assessment of Collembola richness in different sites is required, this eco-morphological trait approach, as well as the determination to the genus level, could be used.     

Defining core habitat of local populations of the gray treefrog (Hyla versicolor) based on choice of oviposition site

Concern over amphibian population declines and loss of terrestrial and aquatic habitat have emphasized the need to define habitat requirements for each stage in a species' life history. The realization that pond-breeding amphibians spend most of their lives in the terrestrial environment suggests the need to protect terrestrial as well as aquatic habitat. Many studies on amphibian populations have focused on emigration from breeding sites to define habitat use; however these studies do not typically elucidate terrestrial activities of adults within the breeding season. We measured colonization rates of artificial pools by gray treefrogs (Hyla versicolor) at multiple distances from natural breeding ponds. We found a non-random distribution of egg deposition among distances, with 95% of eggs deposited within 15 m of the breeding pond. Additionally, we found that the time to first colonization of artificial pools increased with respect to distance. Our results indicate that adult gray treefrogs may travel up to 200 m within a breeding season, and that multiple breeding ponds may be considered part of a single population. We suggest that a minimum core terrestrial habitat of 60 m surrounding breeding sites is appropriate for protection of local populations of gray treefrogs.     

Mapping the Global Emergence of Batrachochytrium dendrobatidis,the Amphibian Chytrid Fungus

The rapid worldwide emergence of the amphibian pathogen Batrachochytrium dendrobatidis (Bd) is having a profound negative impact on biodiversity. However, global research efforts are fragmented and an overarching synthesis of global infection data is lacking. Here, we provide results from a community tool for the compilation of worldwide Bd presence and report on the analyses of data collated over a four-year period. Using this online database, we analysed: 1) spatial and taxonomic patterns of infection, including amphibian families that appear over- and under-infected; 2) relationships between Bd occurrence and declining amphibian species, including associations among Bd occurrence, species richness, and enigmatic population declines; and 3) patterns of environmental correlates with Bd, including climate metrics for all species combined and three families (Hylidae, Bufonidae, Ranidae) separately, at both a global scale and regional (U.S.A.) scale. These associations provide new insights for downscaled hypothesis testing. The pathogen has been detected in 52 of 82 countries in which sampling was reported, and it has been detected in 516 of 1240 (42%) amphibian species. We show that detected Bd infections are related to amphibian biodiversity and locations experiencing rapid enigmatic declines, supporting the hypothesis that greater complexity of amphibian communities increases the likelihood of emergence of infection and transmission of Bd. Using a global model including all sampled species, the odds of Bd detection decreased with increasing temperature range at a site. Further consideration of temperature range, rather than maximum or minimum temperatures, may provide new insights into Bd-host ecology. Whereas caution is necessary when interpreting such a broad global dataset, the use of our pathogen database is helping to inform studies of the epidemiology of Bd, as well as enabling regional, national, and international prioritization of conservation efforts. We provide recommendations for adaptive management to enhance the database utility and relevance.     

Prioritizing conservation areas and vulnerability analyses of the genus Pinus L. (Pinaceae) in Mexico

Mexico hosts the highest species richness of pines (Pinus, Pinaceae) worldwide; however, the priority areas for their conservation in the country are unknown. In this study, the ecological niche of the 50 native pine species was modeled. Then, through a multi-criteria analysis, the priority areas for the conservation of the genus Pinus were identified according to the spatial patterns of richness, geographic rareness, irreplaceability, the level of vulnerability of their habitat and the status of legal protection. The results revealed that the regions with high species richness differed from those with high endemism. Also, most pine species have undergone processes of habitat degradation, having been the endemic species the most affected. The priority areas covered regions with high species richness, high endemism, and highly degraded forests, located at mountainous portions of the Baja California Peninsula, the Sierra Madre Occidental, the Sierra Madre Oriental, the Trans-Mexican Volcanic Belt, and the Sierra Madre del Sur. A low proportion of priority areas overlapped with protected areas or terrestrial regions considered priorities for biological conservation. These results suggest that conservation efforts for this genus should be focused beyond regions with high species richness and current protected areas. Besides, the priority areas identified in this study can be the basis to create biological corridors and new protected areas, which could contribute significantly to the conservation of this genus in Mexico.     

Spatio-ecological niche segregation of two sympatric species of<Emphasis Type="Italic">Clidemia (Melastomataceae)</Emphasis> in Western Amazonian non-flooded rainforests

The frequent occurrence of sympatric series of closely related plant species in tropical rainforests has evoked claims for and against the application of the competitive exclusion principle in these ecosystems. Narrow niche limits defined by biotic as well as abiotic specialization have been reported for sympatric species of the same genus or family. In Amazonian lowland rainforests this question deserves renewed attention because: (1) the existence of edaphically defined community types has recently been well established, and (2) spatio-ecological niche segregation of congeneric species may help explain not only the maintenance of the high Amazonian alpha-diversity, but also its origin through sympatric ecological speciation. In this study, the morphology, ecology, and distribution patterns of two species,Clidemia epiphytica andC. longifolia (Melastomataceae), from western Amazonia, were analyzed. The aims were to find out whether they really are two distinct taxonomic species and if so, whether they also can be considered biological species; if the species are sympatric; and if they are ecologically specialized. The results showed that the morphological variation of the species seems continuous, but that they exhibit opposite morphological responses to variation in soil cation concentration, which suggests that they also are separate biological species. Furthermore, the species occur sympatrically but in different habitats. It is suggested that a part of the enigma of sympatric congeners in rainforests may be explainable by spatial segregation stemming from ecological specialization in relation to subtle environmental variation. It is hypothesized that the studied species are a good candidate case of sympatric speciation driven by ecological specialization.