Separate Species Research Articles

Morphology and phylogenetic position of three anaerobic ciliates from the classes Odontostomatea and Muranotrichea (Ciliophora).

The diversity of the classes Odontostomatea and Muranotrichea, which contain solely obligate anaerobes, is poorly understood. We studied two populations of Mylestoma sp., one of Saprodinium dentatum (Odontostomatea), two of Muranothrix felix sp. nov., and one of Muranothrix sp. (Muranotrichea) employing live observation, protargol impregnation, scanning electron microscopy, and 18S rRNA gene sequencing. Conspecificity of Mylestoma sp., described here, with a previously described species of this genus cannot be excluded since no species have been studied with modern methods. Phylogenetically, the genus Mylestoma is closely related to the odontostomatid Discomorphella pedroeneasi, although the phylogenetic position of class Odontostomatea itself remains unresolved. The newly described muranotrichean species, Muranothrix felix sp. nov., is morphologically similar to M. gubernata but can be distinguished by its fewer macronuclear nodules and fewer adoral membranelles; moreover, it is clearly distinguished from M. gubernata by its 18S rRNA gene sequence. Another population, designated here as Muranothrix sp., most likely represents a separate species.

Problematic putative pachycephalosaurids: Synchrotron µCT imaging shines new light on the anatomy and taxonomic validity of Gravitholus albertae from the Belly River Group (Campanian) of Alberta, Canada

The taxonomic validity of the holotype and sole specimen of the pachycephalosaurid Gravitholus albertae (TMP 1972.027.0001) from the Belly River Group (Alberta, Canada), remains unresolved forty years after its first description. The diagnosis for this species is tenuous at best and extensive cranial fusion has prevented a thorough description and taxonomic referral of TMP 1972.027.0001. We used synchrotron µCT imaging to identify fused sutures and segment the individual elements that comprise TMP 1972.027.0001. This allowed for a detailed description of the specimen in a more thorough comparative framework with other known pachycephalosaurid specimens. Using new observations of contacts between cranial elements, the morphological distinction of TMP 1972.027.0001 from other Belly River Group pachycephalosaurids was tested with bivariate and multivariate morphometric analyses. TMP 1972.027.0001 is morphologically consistent as an end-stage semaphorant of Stegoceras validum. Furthermore, we find no taxonomically significant morphometric distinctions between Gravitholus albertae, Hanssuesia sternbergi, and Stegoceras validum, and propose the former two are synonymous with the later. Large Stegoceras validum frontoparietals show statistically significant dimorphism in the thickness of the frontonasal boss, which is not apparent amongst juvenile and subadult specimens. Pathologies consistent with intraspecific combat (“headbutting”) appear restricted to frontoparietal domes with proportionally taller frontonasal bosses, and suggests that the two morphs represent sexual dimorphs, rather than separate species. Foraminacephale brevis and Stegoceras validum are the only named pachycephalosaurids recognised in the Dinosaur Park Formation. The stratigraphic and temporal range of Stegoceras validum is extended into the underlying Oldman Formation. Pachycephalosaurid diversity in the Campanian is reduced as a result of these revised taxonomic hypotheses. A revised phylogenetic character matrix, recognising taxonomic synonymies and ontogenetically dependent character states results in a largely unresolved Pachycephalosauria.

Crocus speciosus (Iridaceae)—A New Species for the Bulgarian Flora

This is the first report on the autumn-flowering species Crocus speciosus, belonging to C. ser. Speciosi from the Bulgarian flora. The species was found in Southeastern Bulgaria, in the area between Ahtopol and Rezovo. Re-examining the Bulgarian collections, the earliest specimen was collected in 1975, was probably overlooked, and most likely determined as C. pulchellus. The nearest known localities of the species are on the territory of Türkiye. In this study, we compared C. pulchellus and C. ibrahimii using DNA sequence data from the nuclear ITS1/2 region and morphological features. Our study showed a close relationship between the specimens from Bulgaria and the recently deposited data of C. speciosus and their separation from the closely related C. pulchellus. Together with the previously cited white anthers as a key feature for determination, the molecular data confirmed a clear distinction between the samples with white anthers in the two species. The morphological data of our taxon overlapped with the description of C. ibrahimii. The molecular data strongly supported the affiliation of C. speciosus s.l., but did not support the recognition of C. ibrahimii as a separate species and it should be referred to as a subspecies of C. speciosus subsp. ibrahimii Rukšāns.

Identification and reproductive isolation of Euborellia species (Insecta, Dermaptera, Anisolabididae) from East and Southeast Asia.

Euborellia (Anisolabididae: Anisolabidinae) is one of the most speciose genera of earwigs (Dermaptera), and its species-level classification is difficult. To settle the classification of brachypterous species with abbreviated tegmina recorded from East and Southeast Asia, we examined the morphology and reproductive isolation of three tentative Euborellia species, and analyzed the DNA barcoding region of the mitochondrial cytochrome oxidase subunit I (COI) gene. The observed complete reproductive isolation among the three Euborellia taxa and considerable differentiation in the COI sequences clearly show that each should be treated as a separate species. Based on morphology, distribution and the DNA sequence, we identify Euborellia sp. 1 of Malaysia as E.annulata (Fabricius), a circumtropical cosmopolitan with no records of a fully winged form. Samples from Ioto Island (= Iwo-jima Island: Ogasawara Islands, southern Japan) were also identified as this species. Euborellia sp. 3, from the main islands of Japan, was generally larger and lacked a Y-shaped pigmented area on the penis lobe, which is characteristic of Euborellia sp. 1. We propose reinstating E.pallipes (Shiraki) as the oldest name for this taxon. Euborellia sp. 2, even the brachypterous form, can be distinguished from these two species by its paler coloration (particularly the femora), ecarinate post-abdomen, and the shape of the male genitalia (parameres). We tentatively identify this species as E.philippinensis Srivastava based on the morphology of the brachypterous form, although the macropterous form cannot be distinguished from E.femoralis (Dohrn).

Isozyme evidence regarding relationships within Rhizomnium (Mniaceae)

One of the two most speciose genera in the Mniaceae, Rhizomnium includes 14 to 16 named taxa with the likelihood that more remain to be described. Our previous research established that R. pseudopunctatum is an allodiploid whose haploid progenitors are R. gracile and R. magnifolium. In the course of that work, we sampled five additional species as possible progenitors. Using isozymes from starch-gel electrophoresis, we screened all eight species to compare levels of genetic variation within species, genetic distances between species, and genetic population structure. Levels of variation are generally lower than for species of Plagiomnium but are similar to those for species of Cinclidium. Relationships of species based on genetic data are mostly congruent with Koponen's (1973) “assumed phylogeny” of Rhizomnium, but there are also some clear conflicts. For example, R. appalachianum is highly divergent from the other three species placed in sect. Macromnium, and R. glabrescens does not seem to fit in sect. Rhizomnium. Genetic distance data, however, do support Koponen & Sun's (2016) recognition of R. chlorophyllosum as a separate species from R. punctatum.

Taxonomic Notes on an Endemic Variety of Portulaca oleracea [Portulacaceae] Found in India is suppose to Promote as Separate Species Portulaca linearifolia

Santapau & Hennery reported four species of Portulaca found in all over India namely P. oleracea, P. pilosa, P. quadrifida and P. wightiana, among these P. oleracea and P. pilosa further divided in variety and sub species or race by Sivarajan & Manilal and Geesink respectively. In the present investigation author has experienced that P. oleracea var. linearifloia proposed by Sivarajan & Manilal has adequate amount of different characters right now from habit, morphology, shape and size of leaf, shape of operculum and seed testa characteristics hence, assumed to promote the variety as a separate species under name Portulaca linearifolia (Sivarajan & Manilal) Dheeren Panwar com. nov., rather than the variety of type species.

Taxonomic Notes on an Endemic Variety of Portulaca oleracea [Portulacaceae] Found in India is suppose to Promote as Separate Species Portulaca linearifolia

Taxonomic Notes on an Endemic Variety of Portulaca oleracea [Portulacaceae] Found in India is suppose to Promote as Separate Species Portulaca linearifolia

A Revision of the Rhinolophus hipposideros group (Chiroptera: Rhinolophidae) with Definition of an Additional Species from the Middle East

Initially, the Rhinolophus hipposideros group was defined by two morphological traits, the structure of the nose-leaf and the shape of basioccipital bone of the skull. Originally, it consisted of two species, R. hipposideros and R. midas, whereas currently it is considered to contain a single species, R. hipposideros, under whose rank both original species have been joined. The interpretation of geographic variability within the group has traditionally been based on variation in body and skull size, nose-leaf shape, and several selected skull and tooth characters. This approach resulted in delimitations of up to seven subspecies, mostly in the Mediterranean area, a conception introduced more than a hundred years ago and accepted by many authors till today. We investigated the phylogenetic relationships among populations of R. hipposideros with the help of molecular genetic, morphological, and acoustic examinations. Our analysis uncovered the existence of an unexpected diversity within the R. hipposideros group, challenging its current phylogenetic and taxonomic arrangements. The molecular genetic analysis of almost 100 samples and morphological examinations of about 300 specimens showed two main, geographically exclusive, phylogenetic lineages within the group, well delimited by molecular characteristics and possessing two distinct morphotypes and two distinct echotypes. These two lineages are isolated deep enough to be considered separate species. One of them, R. hipposideros s.str., is widespread over the south-western Eurasia and north-western and north-eastern Africa, and the other, R. midas, is distributed in a small range around the Strait of Hormuz and Gulf of Oman. The extensive range of R. hipposideros s.str. is inhabited at least by two subspecies, separated mainly by the genetic characters, whereas the morphological and echolocation traits do not distinguish the populations sufficiently. The western R. h. hipposideros occurs in the Maghreb and Europe west of the Dnieper River, Bosporus, and the Strait of Karpathos, and the eastern R. h. minimus lives east of this boundary, including the populations of Crimea, Caucasus, the Middle East, and north-eastern Africa (Sudan to Djibouti). The two subspecies also differ in karyotype, with 2n = 58 in R. h. minimus and 2n = 54–56 in R. h. hipposideros. The taxonomic position of the easternmost populations of R. hipposideros s.str. (West Turkestan, Afghanistan, Kashmir) remains unresolved and has to be investigated more elaborately and using a more extensive sample set.

A GC-MS and LC-HRMS perspective on the chemotaxonomic investigation of the natural hybrid Origanum × lirium and its parents, O. vulgare subsp. hirtum and O. scabrum.

The genus Origanum L. (Lamiaceae) is widespread in the Mediterranean region. However, approximately 75% of the species are only encountered in the eastern part. Out of these, a total of nine species (11 taxa) and three natural hybrids occur in Greece. Nevertheless, so far, there is no consensus regarding their precise botanical classification in the literature. In fact, the taxon Origanum × lirium has been proposed both as a separate species as well as natural hybrid between Origanum vulgare subsp. hirtum and Origanum scabrum. In this scope, the aim of the current study is to shed light on the matter through the investigation of the chemical composition of both the essential oils and the polar extracts of the mentioned taxa, collected from different geographical regions of Greece. As it was demonstrated by both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-high-resolution mass spectrometry (LC-HRMS) data, and highlighted by our comparative analysis, it can be stipulated that Origanum × lirium shares its chemotype to a large extent with its parent species concerning both volatile and polar constituents. Additionally, geographical origin conditions stood out as a key factor influencing their chemical composition. Altogether, the present work provides useful information on the chemical composition of the taxa under investigation, while our findings support the opinion that Origanum × lirium should be considered not as a separate species, but rather as a hybrid on the way to becoming a species.

Taxonomic and nomenclatural notes on Andryala glandulosa subsp. cheiranthifolia (Cichorieae, Asteraceae) from Madeira Island, Portugal

Currently two subspecies of Andryala glandulosa endemic to Madeira (Portugal) are recognized: A. glandulosa subsp. glandulosa and A. glandulosa subsp. cheiranthifolia. A thorough revision of both taxonomical and nomenclatural aspects led us to the conclusion that the subspecific epithet cheiranthifolia was misapplied. Thus, we suggest to synonymize Andryala glandulosa with A. cheiranthifolia and accept A. glandulosa as the correct name for the taxon presently known as A. glandulosa subsp. glandulosa. The taxon currently recognized as A. glandulosa subsp. cheiranthifolia exhibits distinct vegetative and reproductive structures when compared to A. glandulosa subsp. glandulosa and should, therefore, be treated as a separate species, for which the name A. subglabrata is hereby proposed. A detailed description and illustration of A. subglabrata are provided and its lectotype is designated. Diagnostic characters are discussed and compared with those of A. glandulosa.

New taxonomic status of the Pseudotachycines procerus guizhouensis Zhu & Shi, 2022 (Rhaphidophoridae: Aemodogryllinae: Aemodogryllini)

The genus Pseudotachycines belongs to the tribe Aemodogryllini of the subfamily Aemodogryllinae. In the present paper, we examined the morphological features of male and female adults of the subspecies P. procerus guizhouensis, and suggested that this subspecies should be treated as a separate species. We redescribed the male of P. guizhouensis in detail. The morphological information and some related photographs of the female of this species were provided for the first time in this study. The morphological differentiation between P. guizhouensis and P. procerus was discussed.

In the land of the blind: Exceptional subterranean speciation of cryptic troglobitic spiders of the genus Tegenaria (Araneae: Agelenidae) in Israel

Caves have long been recognized as a window into the mechanisms of diversification and convergent evolution, due to the unique conditions of isolation and life in the dark. These lead to adaptations and reduce dispersal and gene flow, resulting in high levels of speciation and endemism. The Israeli cave arachnofauna remains poorly known, but likely represents a rich assemblage. In a recent survey, we found troglophilic funnel-web spiders of the genus Tegenaria in 26 caves, present mostly at the cave entrance ecological zone. In addition, we identified at least 14 caves inhabited by troglobitic Tegenaria, which are present mostly in the twilight and dark ecological zones. Ten of the caves, located in the north and center of Israel, are inhabited by both troglophilic and troglobitic Tegenaria. These spiders bear superficial phenotypic similarities but differ in the levels of eye reduction and pigmentation. To test whether these taxa constitute separate species, as well as understand their relationships to epigean counterparts, we conducted a broad geographic sampling of cave-dwelling Tegenaria in Israel and Palestine, using morphological and molecular evidence. Counterintuitively, our results show that the troglobitic Tegenaria we studied are distantly related to the troglophilic Tegenaria found at each of the cave entrances we sampled. Moreover, seven new troglobitic species can be identified based on genetic differences, eye reduction level, and features of the female and male genitalia. Our COI analysis suggest that the Israeli troglobitic Tegenaria species are more closely related to eastern-Mediterranean congeners than to the local sympatric troglophile Tegenaria species, suggesting a complex biogeographic history.

Exploring the speciation continuum of slow worms: location and extent of the Anguis fragilis/veronensis hybrid zone in southeastern France

Abstract With five currently recognized species that form several secondary contact zones, slow worms (Anguidae: Anguis) offer a valuable model to study the fate of evolutionary lineages in the face of hybridization and genetic introgression. The relationships between the Western Slow Worm Anguis fragilis and the Italian Slow Worm Anguis veronensis are particularly puzzling. Their respective distributions remain poorly known on the edges of their parapatric ranges, as both species lack external differentiation. Contra earlier mitochondrial phylogenies, new phylogenomic inferences have shown that A. fragilis and A. veronensis are sister taxa, thus casting doubts on their specific status. In this study, we analyze the A. fragilis/veronensis transition in southeastern France, based on one mitochondrial (ND2) and two nuclear (PRLR and HA1) genetic markers in 81 specimens from 61 localities. The ranges of A. fragilis and A. veronensis roughly extends northwest and southeast of the Rhône-Durance valleys, respectively, with clear signs of introgressive hybridization in the areas of contact (notably the eastern parts of the lower Rhône valley). Based on the three molecular markers analyzed, gene flow does not seem to reach outside the narrow hybrid zone, which likely indicates (incomplete) intrinsic reproductive isolation. Hence, we provisionally suggest maintaining A. veronensis as a separate species from A. fragilis. More generally, patterns of genetic divergence, external differentiation, and hybridization (both historical and contemporary) in Anguis ssp. supports a speciation continuum spanning from cryptic, genetically compatible alloparapatric lineages to phenotypically distinct, deeply diverged and fully reproductively isolated taxa able to coexist in sympatry.

Phylogenomic inference and demographic model selection suggest peripatric separation of the cryptic steppe ant species Plagiolepis pyrenaica stat. rev.

The ant Plagiolepis taurica Santschi, 1920 (Hymenoptera, Formicidae) is a typical species of the Eurasian steppes, a large grassland dominated biome that stretches continuously from Central Asia to Eastern Europe and is represented by disjunct outposts also in Central and Western Europe. The extent of this biome has been influenced by the Pleistocene climate, and steppes expanded recurrently during cold stages and contracted in warm stages. Consequently, stenotopic steppe species such as P. taurica repeatedly went through periods of demographic expansion and severe isolation. Here, we explore the impact of these dynamics on the genetic diversification within P. taurica. Delimitation of P. taurica from other Plagiolepis species has been unclear since its initial description, which raised questions on both its classification and its spatiotemporal diversification early on. We re-evaluate species limits and explore underlying mechanisms driving speciation by using an integrative approach based on genomic and morphometric data. We found large intraspecific divergence within P. taurica and resolved geographically coherent western and eastern genetic groups, which likewise differed morphologically. A morphometric survey of type material showed that Plagiolepis from the western group were more similar to P. barbara pyrenaica Emery, 1921 than to P. taurica; we thus lift the former from synonymy and establish it as separate species, P. pyrenaica stat. rev. Explicit evolutionary model testing based on genomic data supported a peripatric speciation for the species pair, probably as a consequence of steppe contraction and isolation during the mid-Pleistocene. We speculate that this scenario could be exemplary for many stenotopic steppe species, given the emphasized dynamics of Eurasian steppes.

Species identification and population genetics of the Antarctic fish genera Lepidonotothen and Nototheniops (Perciformes, Notothenioidei)

AbstractAccurate species identification is essential to assess biodiversity and species richness in ecosystems threatened by rapid and recent environmental changes, such as warming in most Antarctic waters. The Lepidonotothen species complex comprises demersal notothenioid fishes which inhabit the shelf areas of the Antarctic Peninsula, the Scotia Arc and sub‐Antarctic islands with a circum‐Antarctic distribution. Species determination in this group has often been problematic. In particular, whether Lepidonotothen squamifrons and Lepidonotothen kempi are valid as separate species has been questioned. In this study, we analysed the genetic variation among four nominal southern polar species within this complex (L. kempi, L. squamifrons, Nototheniops larseni, Nototheniops nudifrons) by means of three different markers (ND2 and tRNA mitochondrial genes and a panel of 16 nuclear microsatellites). We tested whether individuals morphologically assigned to L. kempi showed genetic separation from L. squamifrons. Our analyses indicated a lack of differentiation between L. kempi and L. squamifrons. However, a genetically distinct population was found for L. squamifrons at the Shag Rocks islands near South Georgia. Antarctic and sub‐Antarctic islands are known to be home to many cryptic species and further studies will elucidate if the genetically differentiated population we found potentially originated from this context and can be considered an incipient species. Our analysis contributes to further characterize the species composition of the most abundant fish suborder in the Southern Ocean, which is among the regions most threatened by climate change.

Challenges and constraints when applying few shot learning to a real-world scenario: In-the-wild camera-trap species classification

Few shot learning (FSL) describes the challenge of learning to classify when there are only few labeled examples. The goal is to be able to adapt to a new classification task using a minimum amount of new data. However, when applying FSL to real-world problems, a number of constraints and challenges are encountered: model selection, training data selection, hyper-parameter and cost function, and final decision making. In this paper, we consider a realistic problem that fits perfectly with the narratives of FSL: our goal is to classify animal species that appear in our in-the-wild camera traps located in Senegal, when these species have yet to appear in popular animal datasets. Using the philosophy of FSL, we first train a FSL network to learn to separate animal species, using popular, large animal datasets, and then evaluate the network with our data, for which there are few labeled images. Using this framework, we conduct a comparison between FSL models, parameter selection, training strategy, etc. We also discuss a weakness of the current FSL framework – lack of the option “does not belong to any of the classes”. This is a common problem in real-world systems due to false positives when detecting bounding boxes that contain, in our case, animals. We then propose an additional constraint to overcome this weakness.

Molecular characterization of Dictyocaulus nematodes in wild red deer Cervus elaphus in two areas of the Italian Alps

Nematodes of the genus Dictyocaulus are the causative agents of parasitic bronchitis and pneumonia in several domestic and wild ungulates. Various species have been described in wild cervids, as the case of Dictyocaulus cervi in red deer, recently described as a separate species from Dictyocaulus eckerti. In Italy, information on dictyocaulosis in wildlife is limited and often outdated. In this work, 250 red deer were examined for the presence of Dictyocaulus spp. in two areas of the Italian Alps (n = 104 from Valle d’Aosta, n = 146 from Stelvio National Park), and the retrieved lungworms were molecularly characterized. Lungworms were identified in 23 and 32 animals from Valle d’Aosta and Stelvio National Park, respectively. The nematodes, morphologically identified as D. cervi, were characterized molecularly (18S rDNA, ITS2, and coxI). Consistently, almost all specimens were found to be phylogenetically related to D. cervi. Three individuals, detected from both study sites and assigned to an undescribed Dictyocaulus sp., clustered with Dictyocaulus specimens isolated from red deer and fallow deer in previous studies. Within each of D. cervi and the undescribed Dictyocaulus sp., the newly isolated nematodes phylogenetically clustered based on their geographical origin. This study revealed the presence of D. cervi in Italian red deer, and an undetermined Dictyocaulus sp. that should be more deeply investigated. The results suggest that further analyses should be focused on population genetics of cervids and their lungworms to assess how they evolved, or co-evolved, throughout time and space and to assess the potential of transmission towards farmed animals.

Additions to the Morphology of the Cucumariidae. I. Intraspecific Variation of the Retractor Muscle in Parathyone braziliensis (Verrill, 1868) and Thyonidium seguroensis (Deichmann, 1930) (Holothuroidea: Dendrochirotida)

Intraspecific variation of the gross morphology of the retractor muscle was studied in 60 specimens of Parathyone braziliensis and 17 specimens of Thyonidium seguroensis. Specimens were dissected to expose the retractor muscle or were analyzed by micro-computed tomography. Early expectations that the point of origin of the retractor muscle at the longitudinal muscle relative to the midpoint of the body length was age-related and that the split of the retractor muscle into separate muscle bands could be used as taxonomic characters to separate species were not supported by anatomical evidence.

Resolving a nearly 90-year-old enigma: The rare Fagus chienii is conspecific with F. hayatae based on molecular and morphological evidence

Taxonomic uncertainties of rare species often hinder effective prioritization for conservation. One such taxonomic uncertainty is the 90-year-old enigma of Fagus chienii. F. chienii was previously only known from the type specimens collected in 1935 in Pingwu County of Sichuan Province, China, and has long been thought to be on the verge of extinction. However, morphological similarities to closely related Fagus species have led many to question the taxonomic status of F. chienii. To clarify this taxonomic uncertainty, we used the newly collected samples to reconstruct a molecular phylogeny of Chinese Fagus species against the phylogenetic backbone of the whole genus using seven nuclear genes. In addition, we examined nine morphological characters to determine whether F. chienii is morphologically distinct from its putatively closest relatives (F. hayatae, F.longipetiolata, and F.lucida). Both morphological and phylogenetic analyses indicated that F. chienii is conspecific with F. hayatae. We recommended that F. chienii should not be treated as a separate species in conservation management. However, conservation strategies such as in situ protection and ex situ germplasm preservation should be adopted to prevent the peculiar “F. chienii” population from extinction.

Genomic evidence for three distinct species in the Erebia manto complex in Central Europe (Lepidoptera, Nymphalidae)

A problem to implement conservation strategies is that in many cases recognized taxa are in fact complexes of several cryptic species. Failure to properly delineate species may lead to misplaced priorities or to inadequate conservation measures. One such species complex is the yellow-spotted ringlet Erebia manto, which comprises several phenotypically distinct lineages, whose degree of genomic isolation has so far not been assessed. Some of these lineages are geographically restricted and thus possibly represent distinct units with conservation priorities. Using several thousand nuclear genomic markers, we evaluated to which degree the bubastis lineage from the Alps and the vogesiaca lineage from the Vosges, are genetically isolated from the widespread manto lineage. Our results suggest that both lineages are genetically as strongly differentiated from manto as other taxonomically well separated sibling species in this genus from each other, supporting a delineation of bubastis and vogesiaca as independent species. Given the restricted and isolated range of vogesiaca as well as the disjunct distribution of bubastis, our findings have significant implication for future conservation efforts on these formerly cryptic species and highlight the need to investigate the genomic identity within species complexes.