Undescribed Species Research Articles

Two new species of Hemidactylus Goldfuss, 1820 (Squamata: Gekkonidae) from the coastal areas of northern Somaliland

The taxonomy of many Hemidactylus geckos from the Horn of Africa has recently been evaluated. However, the lack of fresh material for some species and also regions has led to the misidentification of some taxa and an underestimation of the true diversity in others. In this study we analyse new material of Hemidactylus collected from poorly known coastal areas of northern Somaliland. Our results support the existence of two yet undescribed species within the arid clade. One of the new species is small-sized and closely related to H. afarensis from the Afar Triangle in Ethiopia, and together with two other species (H. foudaii, H. sinaitus) forms the monophyletic group for which we propose here the name sinaitus species group. The other new species is medium to large-sized, known from only one locality, and belongs to the macropholis species group, where it is a sister species of H. macropholis. Both new endemic species from the northern coastal hyper-arid plains highlight the role of northern Somaliland as an important biodiversity hotspot. Furthermore, our results show pronounced genetic diversity within H. macropholis, and imply possible existence of cryptic taxa within the species. Hemidactylus arnoldi and H. tropidolepis have, for the first time, been sequenced and placed in a phylogenetic context in this study. Our results confirm that H. tropidolepis is nested within the African radiation and is most closely related to H. funaiolii. Hemidactylus arnoldi clustered within the Arabian radiation of Hemidactylus and it was recovered as a sister lineage to all other species of the radiation.

Molecular delimitation of cryptic Australian squid species of the genus Uroteuthis Rehder, 1945 (Cephalopoda: Loliginidae), provides a baseline of diversity to resolve classification challenges throughout the Indo-Pacific

ABSTRACT This study provides a comprehensive molecular phylogenetic analysis of Uroteuthis squid from the Indo-Pacific region. The main aim was to increase sample coverage from northern and eastern Australian waters to resolve the identity and distribution of Uroteuthis species taken by local fisheries. Two mitochondrial regions, cytochrome c oxidase subunit I (COI) and 16S ribosomal RNA (16S rRNA), were sequenced from 220 new specimens and analysed with a further 51 sequences from GenBank to create a combined phylogeny for the genus. Three nuclear regions, 18S ribosomal DNA (18S rDNA), 28S ribosomal DNA (28S rDNA) and rhodopsin, were also sequenced from representatives of each species. Based on the mitochondrial phylogeny plus distance and tree-based delimitation models, a COI species barcode gap of 4–5% is proposed for discriminating Uroteuthis species. Applying this gap partitioned many described species into species complexes; for example, U. duvaucelii, U. noctiluca and U. edulis resolved into 10 species. Although more conserved, mitochondrial 16S rRNA sequences differentiated all new species clades, whereas none of the nuclear markers resolved the closest species. Results confirm that neither U. chinensis nor U. edulis occurs in Australian waters. Five undescribed species are identified from northern and eastern Australia, of which four are consistent with earlier allozyme studies (and two align with existing DNA sequences). One is a new southeastern, deeper shelf species differentiated in this study, along with a sixth undescribed species from Indonesian waters. Results of the molecular analysis are now being used to inform complementary morphometric analyses for new species descriptions, and genetic stock structure assessments of these important fisheries resources.

Morphology and molecular evidence reveal hidden diversity among snapping shrimp of the Alpheus obesomanus group (Decapoda: Alpheidae) with the description of a new species from Brazil

The snapping shrimp family Alpheidae Rafinesque, 1815 includes numerous species, most of which present controversial geographical distributions. The disjunct distribution of Alpheus simus Guérin-Méneville, 1856 in the western Atlantic, from Florida to the south of the Caribbean Sea and then from Rio Grande do Norte to Bahia in Brazil, suggests that Brazilian material may belong to an undescribed species. The examination of specimens previously identified as A. simus revealed the distinct morphology of material from Brazil, resulting in the description of a new species based on morphological and molecular evidence. The specimens were collected from fragments of fire coral at the Parque Municipal Marinho do Recife de Fora, Porto Seguro, Bahia, Brazil. Alpheus coralvivo sp. nov. belongs to the A. obesomanus group and is morphologically similar to A. simus (northwestern Atlantic) and A. saxidomus Holthuis, 1980 (eastern Pacific), differing mainly in the stylocerite and scaphocerite lengths and in the large gap between the cephalothorax and pleuron of the first pleonal segment. Molecular analysis using the mitochondrial cytochrome oxidase subunit I gene support the existence of the new species.

A new species of Thamnophis (Serpentes, Colubridae) from Jalisco, Mexico, with a discussion on the phylogeny, taxonomy, and distribution of snakes related to Thamnophis scalaris

Garter snakes in the genus Thamnophis from Mexico have a long and convoluted taxonomic history. From 2015 to 2022, we conducted a comprehensive sampling of Mexican Thamnophis species, aiming to link molecular phylogenies with the recognized species related to T. scalaris in the highlands of Mexico. Here, we present an analysis of mitochondrial DNA to resolve the status of two enigmatic highland Thamnophis populations. Our research resulted in the identification and morphological characterization of a previously undescribed Thamnophis species from the state of Jalisco in western Mexico. We also clarify the identity and relationships of several previously enigmatic populations of Thamnophis. This work presents new data for Thamnophis phylogenetics from the Mexican highlands and offers a framework for future conservation efforts.

Photorhabdus africana sp. nov. isolated from Heterorhabditis entomopathogenic nematodes

One Gram-negative, rod-shaped bacterial strain, isolated from an undescribed Heterorhabditis entomopathogenic nematode species was characterized to determine its taxonomic position. The 16S rRNA gene sequences indicate that it belongs to the class Gammaproteobacteria, to the family Morganellaceae, to the genus Photorhabdus, and likely represents a novel bacterial species. This strain, designated here as CRI-LCT, was therefore molecularly, biochemically, and morphologically characterized to describe the novel bacterial species. Phylogenetic reconstructions using 16S rRNA gene sequences show that CRI-LCT is closely related to P. laumondii subsp. laumondii TT01T and to P. laumondii subsp. clarkei BOJ-47T. The 16rRNA gene sequences between CRI-LCT and P. laumondii subsp. laumondii TT01T are 99.1% identical, and between CRI-LCT and P. laumondii subsp. clarkei BOJ-47T are 99.2% identical. Phylogenetic reconstructions using whole genome sequences show that CRI-LCT is closely related to P. laumondii subsp. laumondii TT01T and to P. laumondii subsp. clarkei BOJ-47T. Moreover, digital DNA-DNA hybridization (dDDH) values between CRI-LCT and its two relative species P. laumondii subsp. laumondii TT01T and P. laumondii subsp. clarkei BOJ-47T are 65% and 63%, respectively. In addition, we observed that average nucleotide identity (ANI) values between CRI-LCT and its two relative species P. laumondii subsp. laumondii TT01T and P. laumondii subsp. clarkei BOJ-47T are 95.8% and 95.5%, respectively. These values are below the 70% dDDH and the 95–96% ANI divergence thresholds that delimits prokaryotic species. Based on these genomic divergence values, and the phylogenomic separation, we conclude that CRI-LCT represents a novel bacterial species, for which we propose the name Photorhabdus africana sp. nov. with CRI-LCT (= CCM 9390T = CCOS 2112T) as the type strain. The following biochemical tests allow to differentiate P. africana sp. nov. CRI-LCT from other species of the genus, including its more closely related taxa: β-Galactosidase, citrate utilization, urease and tryptophan deaminase activities, indole and acetoin production, and glucose and inositol oxidation. Our study contributes to a better understanding of the taxonomy and biodiversity of this important bacterial group with great biotechnological and agricultural potential.

Integrative taxonomy of the featherback fish genus Chitala (Osteoglossomorpha: Notopteridae) in Peninsular Malaysia

AbstractThe taxonomy of the freshwater featherback fish genus Chitala (Osteoglossomorpha: Notopteridae) remains unsettled because delimiting and naming Chitala species based solely on morphology led to different hypotheses. In this study, we aim to determine the number and identity of Chitala species present in Peninsular Malaysia by employing an integrative taxonomic approach that combines morphology and two mitochondrial markers: the cytochrome c oxidase subunit I (COI) and cytochrome b genes. A total of 60 specimens of Chitala were collected throughout Peninsular Malaysia. Initially, all specimens were sorted into three distinct categories based on their body marking patterns. A molecular phylogenetic analysis revealed that each of these groups was monophyletic, and an automated species partition method further recognized them as distinct species. These results support the presence of three Chitala species in Peninsular Malaysia, even though a multivariate morphometric analysis failed to fully differentiate each of these three species. Comparison with original species descriptions and genetic data from an additional 70 Chitala specimens from neighbouring regions allowed us to name these species as Chitala lopis, Chitala borneensis, and Chitala ornata. We also showed the presence of two undescribed species in Indonesia.

A new species of direct-developing frog of the genus Eleutherodactylus (Anura: Eleutherodactylidae) from Tamaulipas, Mexico

The number of species of the subgenus Syrrhophus, genus Eleutherodactylus has increased rapidly in the last eight years, due to recent taxonomic studies. This subgenus of direct-developing frogs is well represented in Mexico, which harbors more than 90% of the species richness reported for the subgenus. In this study, we describe one new species, Eleutherodactylus (Syrrhophus) coelum sp. nov. from the El Cielo Biosphere Reserve in Tamaulipas, Mexico, based on morphological and molecular data. The new species, which was previously assigned to E. cystignathoides, differs from its congeners mainly by its small size and by having the tips of Finger III and IV expanded to twice the width of the narrowest part of these fingers. Phylogenetic analyses recovered a well-supported relationship of the new species as the sister group to an undescribed taxon from San Luis Potosí. This analysis also indicates there are more lineages wating for description.

Development of mitochondrial DNA cytochrome c oxidase subunit I primer sets to construct DNA barcoding library using next-generation sequencing.

Insects are one of the most diverse eukaryotic groups on the planet, with one million or more species present, including those yet undescribed. The DNA barcoding system has been developed, which has aided in the identification of cryptic species and undescribed species. The mitochondrial cytochrome c oxidase I region (mtDNA COI) has been utilised for the barcoding analysis of insect taxa. Thereafter, next-generation sequencing (NGS) technology has been developed, allowing for rapid acquisition of massive amounts of sequence data for genetic analyses. Although NGS-based PCR primers designed to amplify the mtDNA COI region have been developed, their target regions were only a part of COI region and/or there were taxonomic bias for PCR amplification. As the mtDNA COI region is a traditional DNA marker for the DNA barcoding system, modified primers for this region would greatly contribute to taxonomic studies. In this study, we redesigned previously developed PCR primer sets that targetted the mtDNA COI barcoding region to improve amplification efficiency and to enable us to conduct sequencing analysis on NGS. As a result, the redesigned primer sets achieved a high success rate (> 85%) for species examined in this study, covering four insect orders (Coleoptera, Lepidoptera, Orthoptera and Odonata). Thus, by combining the primers with developed primer sets for 12S or 16S rRNA regions, we can conduct more detailed taxonomic, phylogeographic and conservation genetic studies using NGS.

A new antshrike (Aves: Thamnophilidae) endemic to the Caatinga and the role of climate oscillations and drainage shift in shaping cryptic diversity of Neotropical seasonal dry forests

AbstractThe Caatinga is the largest patch of Seasonal Dry Tropical Forest in the Neotropics, located in northeastern Brazil and characterized mainly by deciduous vegetation and extreme rainfall seasonality. It has historically been treated as a biologically impoverished domain, but recent studies uncovered new diversification patterns and several new taxa of frogs, mammals, insects, and fishes. Here we employed a dense sampling regime to evaluate whether the São Francisco River (SFR) would have promoted genetic diversification and fixed phenotypic differences and how Quaternary climatic oscillations shaped distribution and population sizes in a Caatinga endemic species, the Silvery‐cheeked Antshrike (Sakesphoroides cristatus). We adopted an integrative approach using multilocus genetic, plumage, vocal data, and ecological niche modelling (ENM) to characterize evolutionary units and niche suitability in past scenarios. We recovered strong genetic structure across the SFR that was congruent with plumage and vocal variation, revealing a yet undescribed species named herein as Sakesphoroides niedeguidonae, sp. nov. (urn:lsid:zoobank.org:act:2B9FC637‐008A‐4E9A‐B92B‐69ED7FE23823s). The splitting time estimated between the newly described species and S. cristatus is consistent with the establishment of the modern course of SFR, with a more recent course shift apparently promoting the secondary contact between the two species in the Raso da Catarina region. After their split, both species experienced increases in population sizes and range sizes at similar times during the Last Glacial Maximum. We expect other Caatinga avian endemic lineages to show similar patterns of genetic differentiation across the SFR that were enhanced by Quaternary climatic oscillations.

Hidden Phytophthora diversity unveiled in tree nurseries of the Czech Republic with traditional and metabarcoding techniques

Phytophthora diversity was examined in eight forest and ornamental nurseries in the Czech Republic. A leaf baiting isolation technique and, in two nurseries, also Illumina DNA metabarcoding were used to reveal the diversity of Phytophthora in soil and irrigation water and compare the efficacy of both approaches. In total, baiting revealed the occurrence of 12 Phytophthora taxa in 59.4% of soil samples from seven (87.5%) nurseries. Additional baiting of compost was carried out in two nurseries and two Phytophthora species were recovered. Irrigation water was examined in three nurseries by baiting or by direct isolation from partially decomposed floating leaves collected from the water source, and two Phytophthora species were obtained. Illumina sequencing of soil and water samples was done in two and one nurseries, respectively. Phytophthora reads were identified as 45 Phytophthora taxa, 15 of them previously unknown taxa from Clades 6, 7, 8 and 9. Another 11 taxa belonged to known or undescribed species of the oomycete genera Globisporangium, Hyaloperonospora, Nothophytophthora, Peronospora and Plasmopara. Overall, with both techniques 50 Phytophthora taxa were detected with five taxa (P. taxon organica, P. plurivora, P. rosacearum, P. syringae and P. transitoria) being exclusively detected by baiting and 38 only by DNA metabarcoding. Particularly common records in DNA barcoding were P. cinnamomi and P. lateralis which were not isolated by baiting. Only seven species were detected by both techniques. It is recommended to use the combination of both techniques to determine true diversity of Phytophthora in managed or natural ecosystems and reveal the presence of rare or unknown Phytophthora taxa.

Toward a Revision of the Genus Orthosia (Apocynaceae: Asclepiadoideae: Asclepiadeae)

This paper summarizes the present knowledge of the genus Orthosia Decne., based on the study of herbarium material, type specimens, digital images, field observations, and Sanger sequencing of 101 accessions for six chloroplast markers. The results of these analyses encourage a rather narrow species circumscription in most cases. A total of 55 species are recognized, nine of which are new to science. Additionally, incomplete material from some remote areas points to the existence of a handful of yet undescribed species. For some species, morphological and molecular results are incongruent, also indicating the need for future studies. All accepted species are described and illustrated, and their distributions are provided. Nine species and two subspecies are newly described, 13 new combinations and two new synonyms are proposed, and 12 names are lectotypified and one is neotypified.

Notes on an undescribed new Dysphaea species from north-eastern India and north-eastern Bangladesh (Odonata: Euphaeidae)

Notes on an undescribed new Dysphaea species from north-eastern India and north-eastern Bangladesh (Odonata: Euphaeidae)

Recircumscription of the genera Goniozus and Sierola (Hymenoptera: Bethylidae) based on morphological and molecular evidence

A number of undescribed species of the bethylid genera Goniozus and Sierola have been discovered in Australia and North America with character states that had previously been thought to be exclusive to one or the other, prompting a re-examination of the generic boundaries. Analysis of a large dataset of cytochrome oxidase I mtDNA sequences and key morphological characters found that Goniozus is well defined by two insertion-deletion events in COI. The traditional morphological characters can also be used to separate the genera, but variation within the regional fauna must be taken into account. Sierola always has four maxillary palpomeres; Goniozus usually has five, but some Nearctic species have four. Goniozus always has cell 2R1 of the fore wing open, with vein Rs weakly curved; Sierola has Rs bent sharply towards the wing margin, usually closing 2R1, but a small number of recently-described Hawaiian species and many undescribed Australian species and have it narrowly to moderately open. As a result of these changes, two species are transferred from Goniozus to Sierola: Sierola angulata (Muesebeck, 1933) comb. nov. from Australia, and Sierola jamiei (Ward, 2013) comb. nov. from New Zealand.

Hidden invasiveness of non-native Schlegel's Japanese gecko (Reptilia: Squamata: Gekkonidae) and three-way competition among natives and non-natives in Japan.

In this study, we investigated the invasiveness of Gekko japonicus, a prevalent gecko species in Japan and an ancient non-native species, focusing on its competition with both the undescribed endemic Gekko species (referred to as Nishiyamori in Japanese) and G. hokouensis. These species are co-distributed with G. japonicus, leading us to hypothesize that G. japonicus was invasive upon its initial introduction. We employed niche analysis and population genetics through ddRAD-seq to assess the historical invasiveness of G. japonicus by comparing regions with and without interspecies competition. Our niche analysis across the Goto Islands, Hiradojima Island (colonized by G. japonicus) and the Koshikishima Islands (not colonized by G. japonicus) indicated that endemic Gekko sp. alter their microhabitat usage in response to invasions by other gecko species, despite having similar suitable habitats and microhabitat preferences. Population genetic analysis revealed significant population declines in Gekko sp. within areas of introduced competition, in contrast to stable populations in areas without such competition. These findings suggest a tripartite competitive relationship among the gecko species, with G. japonicus and G. hokouensis invasions restricting the distribution of the endemic Gekko sp. Consequently, G. japonicus may have historically acted as an invasive species. Acknowledging the historical dynamics of current biodiversity is crucial for addressing complex ecological issues and making informed conservation decisions.

Ribbon worms (phylum Nemertea) from Bodega Bay, California: A largely undescribed diversity.

The diversity of nemerteans along the Pacific coast of the United States is regarded as well characterized, but there remain many cryptic, undescribed, and "orphan" species (those known only in their larval form). Recent sampling of nemerteans in Oregon and Washington has begun to fill in these taxonomic gaps, but nemertean diversity in California has received relatively little attention over the past 60 years. During the summers of 2019 and 2020, nemertean specimens were collected from 20 locations in the Bodega Bay region of northern California, USA, including rocky intertidal shores, sandy beaches, mudflats, and other habitats. Based on morphological assessment and DNA sequence analysis (partial Cytochrome Oxidase I and 16S rRNA genes), our surveys identified 34 nemertean species. Only 13 of these (38%) can be confidently assigned to described species. Another 11 represent species that are new to science, including members of the genera Riserius, Nipponnemertes, Poseidonemertes, Zygonemertes, Nemertellina, Oerstedia, and three species of uncertain affiliation. The remaining ten species include undescribed or cryptic species of uncertain status that have been found previously along the Pacific Coast of North America. Our surveys also document extensions of known geographic ranges for multiple species, including the first records in California of Antarctonemertesphyllospadicola, Cephalothrixhermaphroditica, and Maculauraoregonensis. This is the first report of the genus Nemertellina in the northeast Pacific and Riserius in California. Overall, our findings highlight how much remains to be learned about the diversity and distribution of nemerteans in the northeast Pacific.

Southern Africa's Great Escarpment as an amphitheater of climate-driven diversification and a buffer against future climate change in bats.

Hosting 1460 plant and 126 vertebrate endemic species, the Great Escarpment (hereafter, Escarpment) forms a semi-circular "amphitheater" of mountains girdling southern Africa from arid west to temperate east. Since arid and temperate biota are usually studied separately, earlier studies overlooked the biogeographical importance of the Escarpment as a whole. Bats disperse more widely than other mammalian taxa, with related species and intraspecific lineages occupying both arid and temperate highlands of the Escarpment, providing an excellent model to address this knowledge gap. We investigated patterns of speciation and micro-endemism from modeled past, present, and future distributions in six clades of southern African bats from three families (Rhinolophidae, Cistugidae, and Vespertilionidae) having different crown ages (Pleistocene to Miocene) and biome affiliations (temperate to arid). We estimated mtDNA relaxed clock dates of key divergence events across the six clades in relation both to biogeographical features and patterns of phenotypic variation in crania, bacula and echolocation calls. In horseshoe bats (Rhinolophidae), both the western and eastern "arms" of the Escarpment have facilitated dispersals from the Afrotropics into southern Africa. Pleistocene and pre-Pleistocene "species pumps" and temperate refugia explained observed patterns of speciation, intraspecific divergence and, in two cases, mtDNA introgression. The Maloti-Drakensberg is a center of micro-endemism for bats, housing three newly described or undescribed species. Vicariance across biogeographic barriers gave rise to 29 micro-endemic species and intraspecific lineages whose distributions were congruent with those identified in other phytogeographic and zoogeographic studies. Although Köppen-Geiger climate models predict a widespread replacement of current temperate ecosystems in southern Africa by tropical or arid ecosystems by 2070-2100, future climate Maxent models for 13 bat species (all but one of those analyzed above) showed minimal range changes in temperate species from the eastern Escarpment by 2070, possibly due to the buffering effect of mountains to climate change.

Two novel Archaeorhizomyces species isolated from ericoid mycorrhizal roots and their association with ericaceous plants in vitro

Archaeorhizomyces is a diverse and ubiquitous genus of the subphylum Taphrinomycotina, which contains soil-inhabiting/root-associated fungi. Although ecological importance and root-associating lifestyles of Archaeorhizomyces can be postulated, morphological aspects of fungal body and root colonization are largely unknown due to the scarcity of cultures. We obtained three unidentified Archaeorhizomyces isolates from ericoid mycorrhizal (ErM) roots of Rhododendron scabrum and Rhododendron × obtusum collected in Japan. To advance our understanding of lifestyle of the genus, we investigated their general morphology, phylogeny, and in vitro root-colonizing ability in ericoid mycorrhizal hosts, Vaccinium virgatum and Rhododendron kaempferi. Some morphological characteristics, such as slow glowing white-to-creamy-colored colonies and formation of yeast-like or chlamydospore-like cells, were shared between our strains and two described species, Archaeorhizomycesfinlayi and Archaeorhizomyces borealis, but they were phylogenetically distant. Our strains were clearly distinguished as two undescribed species based on morphology and phylogenetic relationship. As seen in typical ErM fungi, both species frequently formed hyphal coils within vital rhizodermal cells of ErM plants in vitro. The morphology of hyphal coils was also different between species. Consequently, two novel species, Archaeorhizomyces notokirishimae sp. nov. and Archaeorhizomyces ryukyuensis sp. nov., were described.

DNA barcoding of common freshwater eleotrids (Gobiiformes: Eleotridae) in the Philippines, with a global comparison and notes on their conservation and management

Eleotrids, also known as gudgeons or sleeper gobies, are found mostly in freshwater and mangrove habitats of tropical and subtropical regions. Some eleotrids exhibit minimal morphological variation at the species level. In this study, DNA barcoding was used to assess the molecular diversity of common freshwater eleotrids in the Philippines in relation to other eleotrid species from other countries with available DNA sequences in public databases. A total of 422 mitochondrial DNA cytochrome c oxidase I (COI) gene sequences were analyzed: 107 of which were generated from this study, while the remaining 315 sequences were mined from Barcode of Life Data System and GenBank. Both the neighbor-joining and maximum likelihood trees constructed based on the COI sequences clustered most of the species in concordance with their taxonomic classification. The mean Kimura 2-parameter genetic distances within species and genera were 1.31% and 11.60%, respectively. As expected, the mean interspecific distance was higher than the mean intraspecific distance except in some species. This study revealed the presence of nine distinct eleotrid species belonging to five genera in the Philippines. These species include Giuris laglaizei, G. tolsoni, Eleotris acanthopomus, E. fusca, E. melanosoma, Belobranchus sp., Belobranchus belobranchus, Butis butis, and Ophiocara ophicephalus. This study also revealed the presence of possibly undescribed Belobranchus species in the Philippines. This undescribed Belobranchus species differs by a high percentage of genetic divergence from the other species of Belobranchus (3.00% from B. segura and 13.40% from B. belobranchus). Furthermore, high genetic distance (2.62%) was also observed between G. laglaizei species of Laguna de Bay and Mindanao. This study strongly recommends a taxonomic review of the Laguna de Bay G. laglaizei and the Philippine Belobranchus spp., as these species may harbor species complexes that require closer examination so that proper conservation and management measures can be implemented for each species.

Phylogenetic diversity of the Pristimantis anaiae species group, with a description of a new species of Pristimantis (Anura, Strabomantidae) from Cerro Candelaria, Tungurahua, Ecuador

Continuous field work on mountains in the upper Pastaza watershed, showed an unexpected cryptic diversity in the recently described Pristimantis anaiae species group. Based on molecular and morphological characters we describe a new frog species from the central region of the eastern Ecuadorian Andes. We present an updated phylogeny for the Pristimantis anaiae species group, based on widely used, ubiquitous, and available genes, three mitochondrial genes (12S rRNA, 16S rRNA, ND1) and RAG1 nuclear gene. Our phylogenetic hypothesis increases genetic diversity of the species group and shows two well-supported subclades. We show the relationships of the new species and include for the first time Pristimantis maryanneae, belonging to a clade related to P. resistencia + P. glendae. Our phylogeny shows that new species, from Cerro Candelaria, is more closely related to P. kunam from Sangay National Park and differs from it by having a less conspicuous dorsolateral row of enlarged tubercles, and sacral oval marks surrounded by thin white pale lines. The new species and two additional undescribed species occur on the south side of the Rio Pastaza Valley, at Cerro Candelaria within the Llanganates – Sangay Ecological Corridor, reflecting the high importance of this area for conservation and understanding of evolution processes in the area, suggesting a probably adaptive radiation zone for the P. anaiae species group.

Uncovering further diversity of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) in South American bats

Ochoterenatrema Caballero, 1943 is a genus of lecithodendriid digeneans that prior to this study included 8 species parasitic in bats in the Western Hemisphere. Species of Ochoterenatrema possess a unique morphological feature in form of the pseudogonotyl on the sinistral side of the ventral sucker. In this study, we describe 2 new species of Ochoterenatrema from bats in Ecuador. The new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles, length of oesophagus, sucker ratio and the body shape, among other features. We have generated partial nuclear 28S rDNA and mitochondrial cox1 gene DNA sequences from both new species. The newly obtained sequences were used to differentiate among species and study the phylogenetic interrelationships among Ochoterenatrema spp. The internal topology of the clade was weakly supported, although the cox1 tree was much better resolved than the 28S tree. Comparison of sequences revealed 0–1.2% interspecific divergence in 28S and 3.3–20.5% interspecific divergence in cox1 among Ochoterenatrema spp. The new findings demonstrate that bats in South America likely harbor multiple additional undescribed species of Ochoterenatrema. More extensive sampling from broader geographic and host ranges, especially in North America, should allow for a better understanding of the evolution of host associations and morphological traits of this lineage of lecithodendriid digeneans.