The presence of two morphological forms in the Asian dark firefly Lucidina biplagiata (the Japanese name: Oba-botaru) in Japan was suggested by Dr. Sakyo Kanda in 1935, but they have long been considered to be the morphotypes of a single species. In this study, we examined the genetic variations and temporal emergence of Oba-botaru specimens at a single locality in Aichi, Japan. Phylogenetic analyses of mitochondrial and nuclear markers revealed two distinct lineages within L. biplagiata. Adult emergence patterns differed between the two sympatric groups. We found that the mode of the pronotal color patterns could be largely distinguished between the two groups, and also between bimodal emergence. Preliminary mating experiments showed that males and females from the two genetic groups did not mate in captivity. These results strongly support the existence of two sympatric, reproductively isolated species-level lineages within L. biplagiata.
Abstract Mammals diversified greatly throughout their evolution, giving rise to a wide variety of terrestrial locomotor adaptations and autopodial morphologies. Many extinct mammals exhibit peculiar morphologies, often without extant analogues, whereas others show convergences with phylogenetically distant modern mammals. Xenarthrans exemplify the former pattern; litopterns (extinct), the latter. Xenarthrans comprise three major lineages with distinct morphologies and ecologies: sloths, anteaters, and cingulates. The litopterns included medium-sized ungulates resembling modern horses, along with tridactyl megamammals resembling camels. We used Anatomical Network Analysis to quantify complexity of the manus and pedes of a sample of 31 mammals including extant species and extinct xenarthrans and litopterns, via Principal Component Analysis, Procrustes test, and Partial Least Squares. Digital reduction appears as associated with increased complexity, ranging from autopodia with few highly connected elements to those with many elements but lower connectivity, and a significant, positive covariation between manus and pes. This trend, consistent even in forms with morphofunctionally disparate manus and pedes, indicates remarkable structural resilience in the mammalian autopodia through evolution. Xenarthrans displayed atypical patterns, including marked differences between manus and pes in sloths and anteaters, revealing exceptional autopodial plasticity. Conversely, the autopodial organization of litopterns indicates convergences with modern perissodactyls and artiodactyls.
Abstract Identifying nematode species is challenging due to morphological plasticity and few diagnostic characters, especially in Panagrolaimus, which has many genetically distinct but morphologically similar cryptic species adapted to extreme environments. Morphology-based methods often fail to distinguish divergent Panagrolaimus, limiting discovery. Genome assemblies address these challenges by providing a detailed genetic blueprint for identification. We combined genome sequencing, phylogenetic analyses using 18S rRNA and ultra-conserved elements, and morphology to describe three novel species: Panagrolaimus einhardi sp. nov. (formerly Panagrolaimus sp. ES5, Rhineland, Germany), Panagrolaimus shuimeiren sp. nov. (Namib Desert, Namibia), and Panagrolaimus nebliphilus sp. nov. (Atacama Desert, Chile). Panagrolaimus einhardi is named after Prof. Einhard Schierenberg, a renowned expert and valued member of the nematode community, who isolated the species himself. The identifications are supported by high-quality assemblies from PacBio HiFi or Oxford Nanopore long reads. The P. einhardi assembly was scaffolded using Hi-C, yielding a 116 Mb collapsed assembly with 44 scaffolds (N50: 28 Mb). Panagrolaimus shuimeiren has an assembly size of 69 Mb (49 scaffolds, N50: 13 Mb), and P. nebliphilus an assembly size of 70 Mb (24 scaffolds, N50: 13 Mb). Integrating morphology and genomics clarifies biological diversity in cryptic-rich lineages like Panagrolaimidae, resolving relationships where morphology alone is ambiguous.
Despite the great diversity of the phylum Chordata, lifelong obligate symbiosis with cyanobacteria is only known in certain species belonging to four genera of didemnid ascidians: Didemnum, Diplosoma, Lissoclinum, and Trididemnum. This photosymbiosis in didemnid ascidians exhibits diversity in photosymbiont taxa, symbiont location in the host ascidian, and modes of vertical transmission of symbionts. While the family Didemnidae comprises over 600 colonial species, approximately 30 symbiotic species are known from tropical waters, alongside many non-symbiotic congeners. This study aimed to elucidate the evolutionary history of this unique photosymbiosis in the family Didemnidae by constructing molecular phylogenetic trees based on the partial sequences of two regions in the mitochondrial cytochrome c oxidase subunit I gene (COI) and mapping the character states of photosymbiosis in each ascidian species harboring obligate photosymbionts onto the trees. The molecular phylogenetic trees generally supported the monophyly of Didemnum, Diplosoma, and Lissoclinum, but not that of Trididemnum. In contrast, the phylogenetic tree did not support the monophyly of the following character states of obligate photosymbiosis in ascidians: photosymbiont taxa (Prochloron, Synechocystis, and filamentous species), photosymbiont location (tunic, tunic cells, and peribranchial/common cloacal cavities), and modes of vertical transmission of photosymbionts in which the symbionts are attached to the larval trunk, collected by the rastrum, or transferred by tunic cells. These results suggest that obligate photosymbiosis with similar character states may have been exclusively established multiple times within the family Didemnidae. Because the modes of vertical transmission are closely associated with symbiont location, similar modes of vertical transmission may have evolved multiple times under similar colony structures and reproductive modes.
Abstract Mermithids are a group of nematodes parasitising arthropods and molluscs. Despite their ecological importance and broad host range, they have remained understudied in recent decades. Using molecular taxonomic approaches, we establish 12 new species of mermithids belonging to the genera Mesomermis, Isomermis, and Gastromermis, all isolated from larvae of black flies of the genera Simulium and Prosimulium. We provide an updated phylogeny of mermithids parasitising European representatives of these black fly genera, based on the nuclear 18S rRNA gene, the ITS1–5.8S–ITS2 region, and the mitochondrial cytochrome c oxidase subunit I gene. Secondary structure models of the 18S rRNA, 5.8S rRNA, and ITS2 molecules are also presented. Our aim is to establish a more robust taxonomic framework for this diverse nematode group, as earlier species descriptions relied on poorly preserved juvenile morphology and lacked molecular data, resulting in misidentifications and substantial taxonomic confusion. We demonstrate that molecular methods are highly effective for species delimitation in mermithids, particularly when multiple independent nuclear and mitochondrial loci are combined. The establishment of these new species will support future research on mermithids and help to resolve long-standing taxonomic uncertainties within the family Mermithidae.
Abstract Understanding the processes driving diversification in natural environments requires approaches that can disentangle historical isolation, gene flow, and ecological factors. Homonota underwoodi Kluge, 1964, a gecko endemic to western Argentina, exhibits a broad distribution across the Monte Desert and adjacent highlands, yet its taxonomic boundaries remain unexplored. Here, we used phylogenetic inference, species delimitation, demographic modelling, and morphometric analyses to test for cryptic diversity and to reconstruct the evolutionary history of this taxon. Mitochondrial phylogenies reveal two deeply divergent, geographically structured clades: a widespread lowland lineage and a montane lineage confined to the eastern slopes of the Sierra de Famatina and adjacent systems. Single and multi-locus species delimitation methods consistently support this two-lineage scenario, while demographic modelling favours an isolation-with-migration model with historical bottlenecks in both lineages. Despite limited phenotypic differentiation, environmental association analyses underscore the role of topographic and climatic barriers in maintaining genetic structure. The ‘Famatina’ lineage is strongly supported by delimitation analyses based on genetic data, and its geographic restriction, demographic history, and environmental context provide additional evidence for evolutionary independence. Its description contributes to a growing recognition of the northern Andes and pre-Andean ranges of Argentina as hotspots of diversification shaped by Cenozoic orogeny and climatic shifts.
Abstract While the overwhelming majority of hypotrichous ciliates are lorica-free benthic organisms, certain species like those in the genus Stichotricha Perty, 1849 exhibit dual lifestyles, i.e. residing in self-secreted loricae when attached to substrates, while adopting a free-swimming aloricate existence upon detachment. These chambered ciliates are rarely studied, and most species lack information on their infraciliature, morphogenetic processes, and molecular phylogeny. In this study, a new species, Stichotricha weishanensis sp. nov. and a new Chinese record, also type species, Stichotricha secunda Perty, 1849, were investigated. Morphogenetic information of S. weishanensis sp. nov., which constitutes the first detailed morphogenetic report of this genus, and the first SSU rDNA sequences for both species are provided. Phylogenetic analyses consistently placed Stichotricha within the family Chaetospiridae with full support and revealed a close relationship between Chaetospiridae and Gonostomatidae, supported by the shared Gonostomum-patterned adoral zone, arrangement of dorsal kineties, and certain morphogenetic features. Based on an analysis of the updated information, the diagnoses of Stichotricha and S. secunda are revised and a key to the identification of Stichotricha species is provided. In addition, loricae secretion behaviour, selective adaptive mechanisms and geographic distributions of Stichotricha species are discussed.
Abstract Species defined by striking morphological traits are often identified primarily through these characters, which can obscure subtler variation and lead to underestimation of true diversity and overestimation of geographical ranges of particular species. This issue is evident in the genus Minibiotus, whose species exhibit high morphological similarity and are characterized by long-standing taxonomic uncertainties. Minibiotus aculeatus, originally described by Murray in 1910 and later redescribed by Claxton in 1998, exemplifies this problem. For decades it was identified solely by its conspicuous dorsolateral cuticular spines, regarded as its primary diagnostic characteristic. We examined 19 populations fitting the scarce description of M. aculeatus: 5 from Australia and 14 from New Zealand. Morphological and morphometric traits were analysed using phase contrast and scanning electron microscopy, and molecular analyses were performed on four markers: three nuclear (18S rRNA, 28S rRNA, and ITS-2) and one mitochondrial (COI) DNA fragment. Phylogenetic relationships were reconstructed using Bayesian inference, and species boundaries were estimated with genetic and phenotypic data (both qualitative and quantitative). Our results suggest the presence of not one but ten species, constituting the Minibiotus aculeatus complex. However, the relationships between the species were not fully resolved, resulting in multiple lineages recovered in polytomy. Such a topology indicates that most of the diversification within the complex took place in a short period of time, in comparison to the subsequent evolution of the recovered lineages. Moreover, low levels of congruency between phenotypic and genetic variation of the analysed individuals suggest morphological stasis and/or a young age of the complex, which did not provide sufficient time for the evolution of divergent traits. The stasis and/or insufficient time for morphological differentiation resulted in apparent species crypsis, a phenomenon more and more often integratively documented in various tardigrade groups, which creates serious challenges for delineation and identification of species also in the M. aculeatus complex. Owing to the lack of eggs, we were unable to redescribe M. aculeatus and designate a neotype series and neotype DNA sequences, but we reinstate Minibiotus subjulietae stat. rev. as a valid species. Nevertheless, despite problems with phylogenetic resolution and species delineation, the geographical distribution of the M. aculeatus complex across south-eastern Australasia clearly suggests stenothermic tolerance and preferences for a temperate to cold climate for all species in the complex.
Planigales are the smallest marsupials in the world. Seven species are currently recognized, six that occur in Australia (Planigale ingrami, P. gilesi, P. kendricki, P. maculata, P. tealei and P. tenuirostris) and one that occurs only in New Guinea (P. novaeguineae). Genetic studies using mitochondrial and nuclear DNA fragments have suggested cryptic diversity within both the P. ingrami and P. maculata species complexes that warrant formal taxonomic revision. Here, we addressed the unrecognized diversity in the P. ingrami complex using comprehensive genetic sampling and morphological assessments of specimens from museum collections. We built on previous studies by obtaining DNA sequences from the mitochondrial 12S ribosomal RNA locus and thousands of nuclear single nucleotide polymorphism loci for >220 individuals, comprehensively sampling across species distributions. Phylogenetic analyses and species delimitation approaches were used to define species boundaries and assess divergence between existing and novel species. This was coupled with a thorough morphological examination of museum specimens, including cranial and external morphometrics. We found three diagnosable forms within the P. ingrami species complex. We reinstate Planigale subtilissima, occurring in the Kimberley, from synonymy, synonymize P. ingrami brunnea, and describe a new and highly distinctive species, Planigale petrophila sp. nov., from Arnhem Land in northern Australia.
Abstract In this study, we synthesize the known late Quaternary crocodylian record in Australasia through literature review and direct assessment of fossil and zooarchaeological material. The late Pleistocene record, mainly from Australia, consists of partial skeletal remains mostly referable to the extinct Mekosuchinae, with some attributable to Crocodylus. The youngest reliably dated mekosuchine fossil is ∼20 000 years old, suggesting mekosuchine decline and extinction coincided with that of other Australian megafauna. In contrast, three south-west Pacific islands—New Caledonia, Vanuatu, and Fiji—were Holocene refuges for mekosuchines, whose remains occur in archaeological contexts, indicating human interaction. Their extinction followed soon after human arrival, suggesting anthropogenic influence as a potential key factor. Crocodylus johnstoni occurs in palaeontological (potentially 28 kya) and archaeological sites in Australia. Crocodylus porosus has an ambiguous fossil record potentially going back over 40 kya, but is definitively present by the Holocene. Most Crocodylus remains come from coastal sites overlapping modern ranges. Archaeological evidence supports crocodile utilization by humans in Australia, Torres Strait, and New Guinea. The fragmentary nature of the known fossil material, as well as the current lack of reliable dates, leaves many unanswered questions about the morphology, palaeobiology, and disappearance of mekosuchines.
The emergence of Neogastropoda in the Early Cretaceous and its rapid radiation in the Mid-Cretaceous remain poorly understood due to the extremely limited number of their early fossils. This is why each record from that critically important interval is essential for documenting the early history of the order, as well as for studies utilizing molecular clocks to test hypotheses about evolutionary rates and timing. Here, we describe a new vasid species, Fimbrivasum bulgaricum sp. nov., with a preserved protoconch and a shell morphology with prominent ornamentation and characteristic columellar folds. The specimen also exhibits the earliest occurrences of some functional traits, such as varices and a labral tooth, in Neogastropoda. This finding suggests a much earlier origin of the family Vasidae than previously believed, dating it as the Barremian. It also indicates that neogastropod diversification was well underway in the earliest Cretaceous. We also review the earliest records of living neogastropod superfamilies and their diversification in the Cretaceous showing that the fossil record does not fit well with the molecular phylogeny in the current state of knowledge highlighting the poor fossil record of the group outside of Europe and the United States, and challenges in identification of early members of particular clades.
Abstract Reactive oxygen species (ROS) are crucial signalling molecules, but their excessive accumulation under stress can lead to oxidative damage. In this study, we investigated the influence of age, sex, and duration of anhydrobiosis on the ROS levels in intact tardigrades (Paramacrobiotus experimentalis) and their released coelomocytes (storage cells). The determination of ROS levels, based on confocal microscopy and the quantification of 2,7-dichlorodihydrofluorescein diacetate (DCFH2-DA) fluorescence, was performed 48 h after rehydration following short or long anhydrobiosis (tun duration of 3 or 30 days, respectively), for females and males assigned to three distinct age classes of adult animals (young, mature, and old). ROS levels were influenced differently by the age and sex of animals, in addition to tun duration, and differed distinctly between intact animals and released storage cells, suggesting an important effect of the extracellular environment. Furthermore, although the ROS level could serve as a survival indicator, its predictive power is dependent on age. Thus, the ROS-mediated response to anhydrobiosis is shaped by its duration, in addition to the age and sex of the animal. Moreover, the response differs between cellular and whole-organism levels.
Jumping spiders (Salticidae) are a diverse group of non-web-building predators and the most species-rich family. Rhene is a species-rich genus of the predominantly New World tribe Dendryphantini. However, its monophyly and phylogenetic composition are untested. We present the first molecular phylogeny of Rhene using mitochondrial (COI) and nuclear markers (28S, 18S, and H3), set within a broader framework of the prominent Old World Marpissoida, to test its monophyly and resolve its divergence patterns. Our analyses robustly support the monophyly of Rhene and reveal a previously unrecognised endemic lineage in the montane forests of Sri Lanka that currently comprises two new species: Rhene fuscigera sp. nov. and Rhene nigrigera sp. nov. Additionally, we provide revised descriptions for the Sri Lankan representatives of the widely distributed species R. albigera, R. flavicomans, R. flavigera, and R. rubrigera. Divergence dating indicates an Early Oligocene split (similar to 32.8 Mya) between the Old World (Ballini) and New World (Dendryphantini) lineages of Marpissoida, probably involving transoceanic dispersal. An Early to Middle Miocene dispersal via the Bering land bridge probably facilitated the colonisation of the Old World by dendryphantines. Finally, the endemic Sri Lankan Rhene lineage diversified during the Mid-Pleistocene (similar to 0.63 Mya), consistent with climatic speciation in montane 'sky-island' refugia generated by monsoon-induced habitat fragmentation. The newly described species, with their small population sizes and restricted distribution in montane forests, may be endangered, highlighting the urgent need for conservation assessment.
We erect here the new aeduellid Stephaniella melanocephala gen. et sp. nov. from the Late Carboniferous of the Saint-& Eacute;tienne Basin (Loire, France). This is the first vertebrate to be described in detail from this basin. It is characterized by a unique combination of features such as four extrascapulars, a low maxilla, a trapezoidal suboperculum, two branchiostegal rays, and a sharp caudal inversion. It exhibits a unique large antero-dorsal process of the suboperculum that sets it apart from all other aeduellids. We also redescribe the aeduellid Puertollanichthys ritchiei from the Late Carboniferous of the Puertollano Basin (Spain). This work allows a better reconstruction of the phylogeny of Aeduellidae and confirms several assumptions made in recent studies. Our phylogenetic analysis finds Stephaniella melanocephala Gon & ccedil;alves et al. gen. et sp. nov. as a basal member within Aeduellidae in a clade comprising P. ritchiei and Spinarichthys dispersus. This helps to decipher the early evolutionary history of this family. We highlight that aeduellids underwent an initial radiation in the Stephanian B-C, quickly followed by dispersal events in the northern foreland, hinterland, and southern foreland of the Variscan Belt. Dispersions continued during the Autunian in the Saar-Nahe Basin (Germany, northern foreland).
European subterranean crustaceans represent a significant component of freshwater biodiversity; however, many taxa remain poorly studied. Among them are the groundwater-dwelling isopods of the family Sphaeromatidae, which are distributed across France, Switzerland, Italy, Slovenia, and the Dinaric Karst. We examined specimens spanning the full geographic range of all known species and subspecies, integrating morphological analyses with multilocus DNA data to identify molecular operational taxonomic units (MOTUs) and to test long-standing taxonomic hypotheses. Our results reveal a well-supported clade of European subterranean Sphaeromatidae comprising 29-32 MOTUs, with contrasting patterns of diversity: some morphologically uniform taxa show deep genetic divergence, while others display extensive morphological variation but limited genetic differentiation. The phylogeny supports two geographically distinct genera-Caecosphaeroma in the west and Monolistra in the east-but does not support the traditional subgeneric classification, suggesting paraphyletic or polyphyletic origins. Time-calibrated analyses indicate that the group originated in the Cretaceous, pre-dating the Alpine and Dinaric orogeny, which probably played a major role in subsequent diversification. This study provides the first comprehensive phylogenetic framework for European subterranean Sphaeromatidae, highlights pronounced mismatches between morphological and genetic diversity, and establishes a foundation for future research on their evolution and biogeographic history.
Abstract This study examines the structure of female germ cell clusters and the process of oogenesis in the facultatively parthenogenetic tardigrade Milnesium inceptum Morek, Suzuki, Schill, Georgiev, Yankova, Marley & Michalczyk, 2019 (Eutardigrada, Apochela) using light microscopy, transmission electron microscopy (TEM), and scanning electron microscopy (SEM), as well as 3D reconstructions obtained via serial block-face scanning electron microscopy (SBEM). The meroistic ovary of Mil. inceptum is an unpaired, sac-like structure. It consists of a small germarium with oogonia and a large vitellarium containing germ cell clusters. Each cluster consists of five multinuclear cells interconnected by cytoplasmic bridges and numerous mononuclear cells attached to them. The oogenesis of this species is divided into three phases: previtellogenesis, vitellogenesis, and choriogenesis. The yolk is formed by authosynthesis within both oocytes and trophocytes. The egg capsule consists of two envelopes: a vitelline envelope and a three-layered chorion. The obtained results show that the germ cell cluster organization of Mil. inceptum differs from that described in tardigrades belonging to the order Parachela. These findings provide a deeper understanding of the reproductive biology of tardigrades.
Abstract Tardigrades are well known for their ability to endure desiccation via cryptobiosis, yet their responses to soil moisture variation remain poorly understood in agro-ecosystems. To explore these, we used metabarcoding approaches to investigate tardigrades in 16 pairs of adjacent flooded paddy and non-flooded upland soils, which represent contrasting moisture conditions. Tardigrades were detected in 64 out of 96 samples, with a higher detection rate in paddy soils (88%) compared with upland soils (46%). Paddy soils exhibited significantly higher tardigrade richness and distinct community composition compared to upland soils. Eleven taxonomic lineages were identified in all those samples, with Hypsibius dominating paddy soils and Ursulinius dominating upland soils. Moreover, the proportion of herbivorous tardigrades was significantly higher in paddy soils than in upland soils. Our findings provide evidence that field-management practices and their associated changes in soil biotic and physicochemical properties may critically influence tardigrade richness, community composition, and ecological associations, as well as their contribution to soil food webs. Furthermore, our study underscores that the genera Hypsibius, Grevenius, Acutuncus, Pilatobius, Dianea, and Pseudobiotus are closely associated with soil moisture conditions, suggesting potential sensitivity to altered soil water availability under future climate or land-use change.
Brine shrimp (Artemia) are indispensable in global aquaculture and widely utilized as model organisms across diverse biological disciplines. Nevertheless, a considerable proportion of published studies suffer from irreproducibility and reduced reliability, often due to inadequate consideration of taxonomic resolution. In particular, the persistent and erroneous application of the binomial designation Artemia salina as a generic reference to all species, populations, and parthenogenetic lineages represents a major source of confusion. This article emphasizes that taxonomy is not merely a formal exercise in classical biology but a fundamental scientific framework for cataloguing and conserving Artemia wild biodiversity at both the population/lineage and species levels, in accordance with the International Code of Zoological Nomenclature (ICZN). In the absence of taxonomic verification and clear documentation of the geographic origin of Artemia specimens, the findings of most studies committing this A. salina fallacy cannot be reliably reproduced or independently validated.
Abstract All limno-terrestrial heterotardigrades share a broadly uniform ontogenetic pattern, progressing from two-clawed larvae lacking an anus and gonopore to four-clawed, sexually mature adults through one or two moulting stages. Within the family Echiniscidae, some species undergo changes in the number or length of trunk appendages during development, whereas others exhibit more stable morphologies. Acanthechiniscus is a rarely encountered echiniscid genus characterized by lateral appendages on each trunk segment. Owing to its rarity, detailed ontogenetic data have previously been available only for Acanthechiniscus goedeni. In this study, by integrative analyses of seven Holarctic populations, we characterize the ontogeny of Acanthechiniscus victor and Acanthechiniscus islandicus, and a new species, Acanthechiniscus aestivalis sp. nov., is described based on material collected from North Greenland. Both A. victor and A. aestivalis sp. nov. exhibit stable morphologies across post-embryonic stages, without significant morphometric shifts, whereas A. islandicus and A. goedeni have larvae that differ in chaetotaxy from later stages. In addition to ontogenetic differences, A. victor and A. aestivalis sp. nov. display morphology more similar to Cornechiniscus, whereas A. islandicus and A. goedeni share several distinct phenotypic peculiarities. Phylogenetic analyses based on partial 18S rRNA, 28S rRNA, and COI sequences suggest the diphyly of Acanthechiniscus, with the clade (A. victor and A. aestivalis sp. nov.) most likely sister to Cornechiniscus, based on morphological clues. These findings led to the erection of the Holarctic Polyacanthechiniscus gen. nov., and underline the significance of the Arctic for tardigrade biodiversity research.
Newportia is one of the most diverse genera within the order Scolopendromorpha. Over more than a century, several subgenera have been proposed based mainly on morphology. Few studies to date have used total evidence-based analyses to infer the relationships between the nominal subgenera. The objectives of this work are to use morphological data, and mitochondrial and nuclear molecular markers to study the phylogenetic relationships of these groups and to discuss some of the traditionally recognized morphological characters for the genus. Phylogenetic relationships were inferred using three optimality criteria. The results obtained in all total-evidence analyses provide support for monophyly of the subgenera Tidops and Scolopendrides. The analyses provide evidence for a clade formed by Scolopendrides-like species in which tarsus 2 is not subdivided. Based on the total-evidence analysis we revalidate the subgenus Newportia (Scolopendrides) Saussure, 1858. The monophyly of the subgenus Newportides is not supported in any of the analyses, suggesting that presence or absence of a pretarsal claw on the ultimate leg was subject to homoplasy, but the close relationship of Andeocryptops with Ectonocryptops and Ectonocryptoides was consistently recovered. The recognition of these subgenera will provide a better understanding of phylogenetic relationships within Newportia.
