Taxonomic Sampling Research Articles

Unveiling hidden diversity: Phylogenomics of neotomine rodents and taxonomic implications for the genus Peromyscus.

Neotomine rodents (Cricetidae, Neotominae) represent one of the most commonly encountered and diverse group of rodents in North America, yet phylogenetic relationships within this group remain uncertain. This subfamily is known for its rapid evolution, adding more complexity to our efforts to unravel their evolutionary history. The main debate revolves around the recognition of the genus Peromyscus as monophyletic or paraphyletic due to its relationship with other genera such as Habromys, Megadontomys, Podomys, Neotomodon, and Osgoodomys. Here, we aim to resolve phylogenetic relationships within Neotominae, to further explore their evolutionary history and taxonomic boundaries. We used target capture and high-throughput sequencing of complete mitogenomes and thousands of genome-wide ultraconserved elements loci (UCEs). Our comprehensive analyses encompassed 53 species of Neotominae spanning 12 previously described genera, along with one yet-undescribed genus. We also investigated 12 out of the 13 species groups within Peromyscus. Our analyses including Maximum Likelihood and Bayesian Inference with both mitogemomes and UCEs, as well as the coalescent species-tree-based approach with UCEs, consistently recovered concordant and well-resolved phylogenies with high levels of nodal support. We identified seven main clades within Neotominae that could potentially be recognized at the generic level, mostly to categorize the genus Peromyscus as a monophyletic group, including one species group within "Peromyscus". Furthermore, our divergence dating estimates place the crown age of Neotominae to be around the late Miocene at ca. 7.9 - 10.7 mya. While generic level diversification continued through the Pliocene, species level diversification predominantly occurred during the late Pliocene, extending through the Pleistocene and Holocene. These epochs have been recognized as periods with significant changes in flora and fauna, driving ecological transformations on a global scale. We hypothesized that climatic and vegetation shifts during the Neogene and Quaternary, coupled with geological events, topographical features, and the presence of biogeographical corridors played a pivotal role in the speciation and diversification of Neotominae. Recognizing the importance of generating genomic-scale data coupled with a broad taxonomic sampling, our study, for the first time, offers resolution of the relationships among the main lineages of Neotominae. We expect that the phylogeny presented here will serve as a foundational resource for future systematic and evolutionary studies. This includes facilitating a proper comprehensive taxonomic revision of the group and the formal description and naming of new genera.

Spatial Nonstationarity in Phenological Responses of Nearctic Birds to Climate Variability.

Climate change is shifting the phenology of migratory animals earlier; yet an understanding of how climate change leads to variable shifts across populations, species and communities remains hampered by limited spatial and taxonomic sampling. In this study, we used a hierarchical Bayesian model to analyse 88,965 site-specific arrival dates from 222 bird species over 21 years to investigate the role of temperature, snowpack, precipitation, the El-Niño/Southern Oscillation and the North Atlantic Oscillation on the spring arrival timing of Nearctic birds. Interannual variation in bird arrival on breeding grounds was most strongly explained by temperature and snowpack, and less strongly by precipitation and climate oscillations. Sensitivity of arrival timing to climatic variation exhibited spatial nonstationarity, being highly variable within and across species. A high degree of heterogeneity in phenological sensitivity suggests diverging responses to ongoing climatic changes at the population, species and community scale, with potentially negative demographic and ecological consequences.

Insect Mitochondrial Genomics: A Decade of Progress.

The past decade has seen the availability of insect genomic data explode, with mitochondrial (mt) genome data seeing the greatest growth. The widespread adoption of next-generation sequencing has solved many earlier methodological limitations, allowing the routine sequencing of whole mt genomes, including from degraded or museum specimens and in parallel to nuclear genomic projects. The diversity of available taxa now allows finer-scale comparisons between mt and nuclear phylogenomic analyses; high levels of congruence have been found for most orders, with some significant exceptions (e.g., Odonata, Mantodea, Diptera). The evolution of mt gene rearrangements and their association with haplodiploidy have been tested with expanded taxonomic sampling, and earlier proposed trends have been largely supported. Multiple model systems have been developed based on findings unique to insects, including mt genome fragmentation (lice and relatives) and control region duplication (thrips), allowing testing of hypothesized evolutionary drivers of these aberrant genomic phenomena. Finally, emerging research topics consider the contributions of mt genomes to insect speciation and habitat adaption, with very broad potential impacts. Integration between insect mt genomic research and other fields within entomology continues to be our field's greatest opportunity and challenge.

A targeted gene phylogenetic framework to investigate diversification in the highly diverse yet geographically restricted red devil spiders (Araneae, Dysderidae).

The family Dysderidae is a highly diverse group of nocturnal ground-dwelling and active-hunter spiders. Dysderids are mostly restricted to the Western Palearctic, and particularly rich and abundant around the Mediterranean region. Interestingly, the distribution of species richness among its 24 genera and three subfamilies is highly biased-80% of its 644 documented species belong to just two genera, Dysdera (326) and Harpactea (211). Dysderidae provides an excellent study case for evolutionary and ecological research. It includes cases of trophic specialization, which are uncommon among spiders, and exhibit other remarkable biological (e.g. holocentric chromosomes), behavioural (e.g. cryptic female choice), evolutionary (e.g. adaptive radiation) and ecological features (e.g. recurrent colonization of the subterranean environment). The lack of a quantitative hypothesis on its phylogenetic structure has hampered its potential as a testing ground for evolutionary, biogeographical and ecological hypotheses. Here, we present the results of a target, multi-locus phylogenetic analysis, using mitochondrial (cox1, 16s and 12s) and nuclear genes (h3, 28s and 18s), of the most exhaustive taxonomic sample within Dysderidae (104 spp.) to date and across related families (Synspermiata) (83 spp.). We estimate divergence times using a combination of fossil and biogeographic node calibrations and use this timeline to identify shifts in diversification rates. Our results support the monophyly of the Dysderidae subfamilies Rhodinae and Dysderinae but reject Harpacteinae as currently defined. Moreover, the clades recovered within Harpacteinae do not support its current taxonomy. The origin of the family most likely post-dated the break-up of Pangea, and cave colonization may be older than previously considered. After correcting for the taxonomic artefacts, we identified a significant shift in diversification rates at the base of the genus Dysdera. Although the unique coexistence of specialist and generalist diets within the lineage could be suggested as the potential driver for the rate acceleration, further quantitative analyses would be necessary to test this hypothesis.

Underground speciation: Unraveling the systematics and evolution of the highly diverse tuco-tucos (genus Ctenomys) with genomic data

Subterranean rodents of the genus Ctenomys (tuco-tucos) are endemic to South America and have experienced relatively recent radiation. There are about 67 recognized species that originated in approximately 1–2 MY. They stand out for their species richness, extraordinary chromosomal diversity, and wide range of habitat they occupy in the continent. Phylogenetic relationships among species of tuco-tucos have been challenging to resolve. Groups of closely-related species have been suggested, but their relationships must be resolved. This study estimates the phylogeny of the genus using massive sequencing, generating thousands of independent molecular markers obtained by RADseq, with a taxonomic sampling that includes 66% of the recognized species. The sequences obtained were mapped against the C. sociabilis genome, recovering up to 1,215 widely shared RAD loci with more than 19,000 polymorphic sites. Our new phylogenetic hypothesis corroborated the species groups previously proposed with cytochrome b gene sequences and provided a much greater resolution of the relationships among species groups. The frater group is sister to all other tuco-tucos, whereas some of the earlierliest proposals placed the sociabilis group as sister to all other tuco-tucos. Ctenomys leucodon, previously proposed as an independent lineage, is associated with the frater group with moderate statistical support. The magellanicus and mendocinus are sister groups in a major clade formed by the boliviensis, talarum, tucumanus, torquatus, and opimus groups. Ctenomys viperinus, included in the phylogeny for the first time, belongs to the tucumanus group. This multi-locus phylogenetic hypothesis provides insights into the historical biogeography of understanding this highly diverse genus.

Allometry and phylogenetic divergence: Correspondence or incongruence?

The potential connection between trends of within species variation, such as those of allometric change in morphology, and phylogenetic divergence has been a central topic in evolutionary biology for more than a century, including in the context of human evolution. In this study, I focus on size-related shape change in craniofacial proportions using a sample of more than 3200 adult Old World monkeys belonging to 78 species, of which 2942 specimens of 51 species are selected for the analysis. Using geometric morphometrics, I assess whether the divergence in the direction of static allometries increases in relation to phyletic differences. Because both small samples and taxonomic sampling may bias the results, I explore the sensitivity of the main analyses to the inclusion of more or less taxa depending on the choice of a threshold for the minimum sample size of a species. To better understand the impact of sampling error, I also use randomized subsampling experiments in the largest species samples. The study shows that static allometries vary broadly in directions without any evident phylogenetic signal. This variation is much larger than previously found in ontogenetic trajectories of Old World monkeys, but the conclusion of no congruence with phylogenetic divergence is the same. Yet, the effect of sampling error clearly contributes to inaccuracies and tends to magnify the differences in allometric change. Thus, morphometric research at the boundary between micro- and macro-evolution in primates, and more generally in mammals, critically needs very large and representative samples. Besides sampling error, I suggest other non-mutually exclusive explanations for the lack of correspondence between allometric and phylogenetic divergence in Old World monkeys, and also discuss why directions might be more variable in static compared to ontogenetic trajectories. Even if allometric variation may be a poor source of information in relation to phylogeny, the evolution of allometry is a fascinating subject and the study of size-related shape changes remains a fundamental piece of the puzzle to understand morphological variation within and between species in primates and other animals.

The moulting arthropod: a complete genetic toolkit review.

Exoskeletons are a defining character of all arthropods that provide physical support for their segmented bodies and appendages as well as protection from the environment and predation. This ubiquitous yet evolutionarily variable feature has been instrumental in facilitating the adoption of a variety of lifestyles and the exploitation of ecological niches across all environments. Throughout the radiation that produced the more than one million described modern species, adaptability afforded by segmentation and exoskeletons has led to a diversity that is unrivalled amongst animals. However, because of the limited extensibility of exoskeleton chitin and cuticle components, they must be periodically shed and replaced with new larger ones, notably to accommodate the growing individuals encased within. Therefore, arthropods grow discontinuously by undergoing periodic moulting events, which follow a series of steps from the preparatory pre-moult phase to ecdysis itself and post-moult maturation of new exoskeletons. Each event represents a particularly vulnerable period in an arthropod's life cycle, so processes must be tightly regulated and meticulously executed to ensure successful transitions for normal growth and development. Decades of research in representative arthropods provide a foundation of understanding of the mechanisms involved. Building on this, studies continue to develop and test hypotheses on the presence and function of molecular components, including neuropeptides, hormones, and receptors, as well as the so-called early, late, and fate genes, across arthropod diversity. Here, we review the literature to develop a comprehensive overview of the status of accumulated knowledge of the genetic toolkit governing arthropod moulting. From biosynthesis and regulation of ecdysteroid and sesquiterpenoid hormones, to factors involved in hormonal stimulation responses and exoskeleton remodelling, we identify commonalities and differences, as well as highlighting major knowledge gaps, across arthropod groups. We examine the available evidence supporting current models of how components operate together to prepare for, execute, and recover from ecdysis, comparing reports from Chelicerata, Myriapoda, Crustacea, and Hexapoda. Evidence is generally highly taxonomically imbalanced, with most reports based on insect study systems. Biases are also evident in research on different moulting phases and processes, with the early triggers and late effectors generally being the least well explored. Our synthesis contrasts knowledge based on reported observations with reasonably plausible assumptions given current taxonomic sampling, and exposes weak assumptions or major gaps that need addressing. Encouragingly, advances in genomics are driving a diversification of tractable study systems by facilitating the cataloguing of putative genetic toolkits in previously under-explored taxa. Analysis of genome and transcriptome data supported by experimental investigations have validated the presence of an "ultra-conserved" core of arthropod genes involved in moulting processes. The molecular machinery has likely evolved with elaborations on this conserved pathway backbone, but more taxonomic exploration is needed to characterise lineage-specific changes and novelties. Furthermore, linking these to transformative innovations in moulting processes across Arthropoda remains hampered by knowledge gaps and hypotheses based on untested assumptions. Promisingly however, emerging from the synthesis is a framework that highlights research avenues from the underlying genetics to the dynamic molecular biology through to the complex physiology of moulting.

Exploring Phylogenetic Relationships of the Mecocephala Group (Hemiptera: Pentatomidae) Using Continuous and Discrete Characters, with Notes on Taxonomy.

The Mecocephala group comprises about 50 species, restricted to the Neotropics and with the highest species richness in Neotropical South America. Several species use rice as host plants and their identification is facilitated by the presence of exaggerated head proportions and a unique male genitalic morphology. The taxonomy of the group has been extensively explored, but inferring its monophyly and especially its internal phylogenetic relationships has been challenging. Here, we inferred the phylogenetic relationships for the group assembling the most complete taxonomic sampling to date, analyzing discrete and continuous morphological characters through equal and implied weighted parsimony analyses. The monophyly of the group was recovered, but internal relationships varied slightly according to the dataset tested. Thus, we propose internal arrangements for the group and provide a formal description of the Mecocephala group, diagnoses for each genus, a dichotomous key to identify its genera, and illustrations of the morphological characters and type species.

Phylogeography of mammals in Southeast Alaska and implications for management of the Tongass National Forest

AbstractInsular evolution on archipelagos generates a significant proportion of global biodiversity, yet islands are among the ecosystems most sensitive to accelerating anthropogenic disturbance, introductions of non‐native species, and emerging pathogens, among other conservation challenges. The Alexander and Haida Gwaii archipelagos along North America's North Pacific Coast support a disproportionate number of endemic taxa compared to other high‐latitude terrestrial ecosystems. In this region, endemics in Canada are explicitly protected, but in the United States, endemics have been operationally ignored. We reviewed regional research on terrestrial mammals and endemics from 2000–2022 to guide wildlife management. Elevated regional endemism is due to a combination of deep and shallow temporal processes (i.e., long‐term refugial isolation vs. recent colonization). With adequate sampling, genomic analyses are well‐suited to identifying nuanced patterns of divergence and endemism, thereby facilitating a deeper understanding of regional diversity. We identified 18 mammalian endemics in Southeast Alaska, USA, at varying taxonomic scales, but research effort has significant taxonomic biases and sampling infrastructure remains inadequate. Of the 66 terrestrial and aquatic mammal species in Southeast Alaska, only 55% are represented by ≥10 archived samples over the last 2 decades. Across taxa, major spatial and temporal sampling gaps limit interpretations of wildlife responses to changing environmental conditions. The Tongass National Forest is spread across an island archipelago, and climate change is projected to have disproportionate impacts on island endemics worldwide. In this case, the United States Forest Service is not closely monitoring endemic taxa, as was required by the Tongass Land Management Plan in 1997. Our review underscores a need for increased consideration of how endemism can be incorporated into land and wildlife management across the Alexander Archipelago. Moving forward, we encourage state and federal agencies, Indigenous communities, and international collaborators to continue to partner with natural history biorepositories to ensure strategic wildlife sampling infrastructure is built and made accessible to the broader scientific community as part of the land management process.

Complete phylogeny of Micrathena spiders suggests multiple dispersal events among Neotropical rainforests, islands and landmasses, and indicates that Andean orogeny promotes speciation.

The Neotropical region is the most diverse on the planet, largely owing to its mosaic of tropical rainforests. Multiple tectonic and climatic processes have been hypothesized to contribute to generating this diversity, including Andean orogeny, the closure of the Isthmus of Panama, the GAARlandia land bridge and historical connections among currently isolated forests. Micrathena spiders are diverse and widespread in the region, and thus a complete phylogeny of this genus allows the testing of hypotheses at multiple scales. We estimated a complete, dated phylogeny using morphological data for 117 Micrathena species and molecular data of up to five genes for a subset of 79 species. Employing event-based approaches and biogeographic stochastic mapping while considering phylogenetic uncertainty, we estimated ancestral distributions, the timing and direction of dispersal events and diversification rates among areas. The phylogeny is generally robust, with uncertainty in the position of some of the species lacking sequences. Micrathena started diversifying around 25 Ma. Andean cloud forests show the highest in-situ speciation, while the Amazon is the major dispersal source for adjacent areas. The Dry Diagonal generated few species and is a sink of diversity. Species exchange between Central and South America involved approximately 23 dispersal events and started ~20 Ma, which is consistent with a Miocene age for the Isthmus of Panama closure. We inferred four dispersal events from Central America to the Antilles in the last 20 Myr, indicating the spiders did not reach the islands through the GAARlandia land bridge. We identified important species exchange routes among the Amazon, Andean cloud forests and Atlantic forests during the Plio-Pleistocene. Sampling all species of the genus was fundamental to the conclusions above, especially in identifying the Andean forests as the area that generated the majority of species. This highlights the importance of complete taxonomic sampling in biogeographic studies.

Unifying functional and population ecology to test the adaptive value of traits.

Plant strategies are phenotypes shaped by natural selection that enable populations to persist in a given environment. Plant strategy theory is essential for understanding the assembly of plant communities, predicting plant responses to climate change, and enhancing the restoration of our degrading biosphere. However, models of plant strategies vary widely and have tended to emphasize either functional traits or life-history traits at the expense of integrating both into a general framework to improve our ecological and evolutionary understanding of plant form and function. Advancing our understanding of plant strategies will require investment in two complementary research agendas that together will unify functional ecology and population ecology. First, we must determine what is phenotypically possible by quantifying the dimensionality of plant traits. This step requires dense taxonomic sampling of traits on species representing the broad diversity of phylogenetic clades, environmental gradients, and geographical regions found across Earth. It is important that we continue to sample traits locally and share data globally to fill biased gaps in trait databases. Second, we must test the power of traits for explaining species distributions, demographic rates, and population growth rates across gradients of resource limitation, disturbance regimes, temperature, vegetation density, and frequencies of other strategies. This step requires thoughtful, theory-driven empiricism. Reciprocal transplant experiments beyond the native range and synthetic demographic modelling are the most powerful methods to determine how trait-by-environment interactions influence fitness. Moving beyond easy-to-measure traits and evaluating the traits that are under the strongest ecological selection within different environmental contexts will improve our understanding of plant adaptations. Plant strategy theory is poised to (i) unpack the multiple dimensions of productivity and disturbance gradients and differentiate adaptations to climate and resource limitation from adaptations to disturbance, (ii) distinguish between the fundamental and realized niches of phenotypes, and (iii) articulate the distinctions and relationships between functional traits and life-history traits.

Please Mind the Gap: Indel-Aware Parsimony for Fast and Accurate Ancestral Sequence Reconstruction and Multiple Sequence Alignment Including Long Indels.

Despite having important biological implications, insertion, and deletion (indel) events are often disregarded or mishandled during phylogenetic inference. In multiple sequence alignment, indels are represented as gaps and are estimated without considering the distinct evolutionary history of insertions and deletions. Consequently, indels are usually excluded from subsequent inference steps, such as ancestral sequence reconstruction and phylogenetic tree search. Here, we introduce indel-aware parsimony (indelMaP), a novel way to treat gaps under the parsimony criterion by considering insertions and deletions as separate evolutionary events and accounting for long indels. By identifying the precise location of an evolutionary event on the tree, we can separate overlapping indel events and use affine gap penalties for long indel modeling. Our indel-aware approach harnesses the phylogenetic signal from indels, including them into all inference stages. Validation and comparison to state-of-the-art inference tools on simulated data show that indelMaP is most suitable for densely sampled datasets with closely to moderately related sequences, where it can reach alignment quality comparable to probabilistic methods and accurately infer ancestral sequences, including indel patterns. Due to its remarkable speed, our method is well suited for epidemiological datasets, eliminating the need for downsampling and enabling the exploitation of the additional information provided by dense taxonomic sampling. Moreover, indelMaP offers new insights into the indel patterns of biologically significant sequences and advances our understanding of genetic variability by considering gaps as crucial evolutionary signals rather than mere artefacts.

An evaluation of fish and invertebrate mercury concentrations in the Caribbean Region

Mercury is a ubiquitous pollutant of global concern but the threat of exposure is not homogenously distributed at local, regional, or global scales. The primary route of human exposure to mercury is through consumption of aquatic foods, which are culturally and economically important in the wider Caribbean Region, especially for Small Island Developing States (SIDS). We compiled more than 1600 samples of 108 unique species of fish and aquatic invertebrates collected between 2005 and 2023 from eleven countries or territories in the wider Caribbean Region. There was wide variability in total mercury concentrations with 55% of samples below the 0.23 µg/g wet weight (ww) guideline from the U.S. FDA/EPA (2022) for 2 or 3 weekly servings and 26% exceeding the 0.46 µg/g ww guideline consistent with adverse effects on human health from continual consumption, particularly for sensitive populations. Significant relationships were found between total mercury concentrations and taxonomic family, sampling country, fish length, and trophic level. The data analyzed here support the need for further sampling with concrete geospatial data to better understand patterns and mechanisms in mercury concentrations and allow for more informed decision making on the consumption of fish and invertebrates from the wider Caribbean Region as well as supporting efforts to evaluate the effectiveness of national, regional, and international mercury policies.

DeepDive: estimating global biodiversity patterns through time using deep learning

Understanding how biodiversity has changed through time is a central goal of evolutionary biology. However, estimates of past biodiversity are challenged by the inherent incompleteness of the fossil record, even when state-of-the-art statistical methods are applied to adjust estimates while correcting for sampling biases. Here we develop an approach based on stochastic simulations of biodiversity and a deep learning model to infer richness at global or regional scales through time while incorporating spatial, temporal and taxonomic sampling variation. Our method outperforms alternative approaches across simulated datasets, especially at large spatial scales, providing robust palaeodiversity estimates under a wide range of preservation scenarios. We apply our method on two empirical datasets of different taxonomic and temporal scope: the Permian-Triassic record of marine animals and the Cenozoic evolution of proboscideans. Our estimates provide a revised quantitative assessment of two mass extinctions in the marine record and reveal rapid diversification of proboscideans following their expansion out of Africa and a >70% diversity drop in the Pleistocene.

Mitochondrial genome rearrangements and phylogenomics of the Hymenoptera (Insecta) using an expanded taxon sample

The order Hymenoptera is one of the most species-rich insect orders, with more than 150,000 described extant species. Many hymenopteran insects have very different mitochondrial genome (mitogenome) organizations compared to the putative ancestral organization of insects. In this study, we sequenced 18 mitogenomes of representatives in the order Hymenoptera to increase taxonomic sampling. A total of 475 species were used in phylogenetic analyses, including 18 new mitogenomes and 457 existing mitogenomes. Using a site-heterogeneous model, Bayesian’s inference from amino acid data yielded more resolved relationships among Hymenoptera than maximum-likelihood analysis and coalescent-based species analyses. The monophyly of Symphyta was not supported. The Xyeloidea was the earliest branching clade in the Hymenoptera. The Orussoidea was closely related to Apocrita. Within Apocrita, the Parasitoida was non-monophyletic. The monophyly of most Parasitoida superfamilies received strong support. The Proctotrupomorpha clade was supported in Bayesian’s analysis. The Apoidea was monophyletic when excluding Ampulex compressa from consideration. The superfamilies Vespoidea and Chrysidoidea were found to be non-monophyletic. Comparisons of mitochondrial gene order revealed a higher frequency of gene rearrangement among lineages with a parasitoid lifestyle, particularly prominent in Chalcidoidea. The degree of gene rearrangement ranked second in specific taxa of Cynipoidea and Ichneumonoidea.

Diversity and biogeography of South American mud turtles elucidated by multilocus DNA sequencing (Testudines: Kinosternidae)

Kinosternon is the most speciose genus of extant turtles, with 22 currently recognized species, distributed across large parts of the Americas. Most species have small distributions, but K. leucostomum and K. scorpioides range from Mexico to South America. Previous studies have found discordance between mitochondrial and nuclear phylogenies in some kinosternid groups, with the current taxonomy following the nuclear-based results. Herein, based on extended molecular, geographic, and taxonomic sampling, we explore the phylogeographic structure and taxonomic limits for K. leucostomum and the K. scorpioides group and present a fossil-calibrated nuclear time tree for Kinosternon. Our results reveal contrasting differentiation patterns for the K. scorpioides group and K. leucostomum, despite overlapping distributions. Kinosternon leucostomum shows only shallow geographic divergence, whereas the K. scorpioides group is polyphyletic with up to 10 distinct taxa, some of them undescribed. We support the elevation of K. s. albogulare and K. s. cruentatum to species level. Given the deep divergence within the genus Kinosternon, we propose the recognition of three subgenera, Kinosternon, Cryptochelys and Thyrosternum, and the abandonment of the group-based classification, at least for the K. leucostomum and K. scorpioides groups. Our results show an initial split in Kinosternon that gave rise to two main radiations, one Nearctic and one mainly Neotropical. Most speciation events in Kinosternon occurred during the Quaternary and we hypothesize that they were mediated by both climatic and geological events. Additionally, our data imply that at least three South American colonizations occurred, two in the K. leucostomum group, and one in the K. scorpioides group. Additionally, we hypothesize that discordance between mitochondrial and nuclear phylogenetic signal is due to mitochondrial capture from an extinct kinosternine lineage.

Comparison of DNA metabarcoding and microscopy in analysing planktonic protists from the European Arctic

In the era of climate change-related restructuring of planktonic protist communities, it is especially important to identify possible shifts in their taxonomic composition. While traditional microscopy-based morphological classification is time-consuming and requires experienced taxonomists, metabarcoding seems to substantially accelerate the determination of taxonomic composition. In this study, based on samples collected in summer 2019 from the West Spitsbergen Current, we analysed planktonic protists using both methods. Metabarcoding, based on high-throughput sequencing of the V4 region of the 18S rRNA gene, resulted in a much higher number of operational taxonomic units (OTUs) and sample diversity than microscopy, although the resolution of taxonomic identification ranged from species to phyla. Most morphology-based identification was performed at the species or genus level, additionally allowing us to include information about dominants and size fractions. The highest proportion of 45% shared taxa by both methods was recorded at the class level. The composition of dominant protists differed between the approaches, with most similarities being observed in Bacillariophyceae, for which two genera, Thalassiosira and Eucampia, were found to be the most abundant with both methods. For Dinophyceae, the most abundant representatives identified by microscopy were Gymnodinium spp., Prorocentrum minimum and Gonyaulax gracilis, while in the metabarcoding approach, most dinoflagellates were identified to the class level only. Given the different levels of accuracy of taxonomic determinations and possible biases in results connected to the chosen methodology, we advocate using an integrative taxonomic approach for the classification of planktonic protists based on the combination of microscopy and molecular methods.

Phylogeny, chronology, and phylogeography in Australasian Hypnodendraceae

Abstract The Hypnodendrales form the sister group to all other pleurcarpous mosses in the superorder Hypnanae. Many species are closely associated with the cool temperate rainforests of Australasia, while others are widespread in higher altitude wet forests across tropical South-East Asia. Several of the Malesian entities are taxonomically ambiguous, having been variously treated as species complexes or as single very variable species. Here we use dated phylogenies to examine the timing of diversification of major clades within the Hypnodendrales, test the hypothesis that widespread, taxonomically problematic species have diversified relatively recently in South-East Asia from within clades of southern temperate origin, and address taxonomic questions within Hypnodendron. A chronogram with broad taxonomic sampling is constructed, followed by a second dated phylogeny with dense sampling from Hypnodendron vitiense, a representative morphologically variable species found throughout Eastern Australia and South-East Asia. The crown group Hypnodendrales are found to have originated in the mid-Cretaceous, although they share a Most Recent Common Ancestor (MRCA) with the other Hypnanae in the late Jurassic. Extant members of Hypnodendron share a MRCA ±28 Mya, a little before the start of the collision of Australia with the Sunda plate that initiated the Australasian–Malesian floristic interchange. Within H. vitiense, there is strong phylogenetic structure consistent with the diversification and isolation of populations in South-East Asia within the last 10 Mya. The New Zealand endemic H. marginatum is found to be derived from within an Australasian clade of H. vitiense, this in turn being distinct from a South-East Asian/tropical Australian H. vitiense clade. Our results suggest that the phylogeography of this prominent group of mosses closely mirrors that of the rainforest ecosystems of which they are a part.

New dinosaur remains increase theropod diversity in the Cañadón Asfalto Formation (Lower Jurassic), Chubut Province, Argentina

The Jurassic record of South American theropods is scarce, being represented by only seven nominal species. Four of these species (Eoabelisaurus mefi, Asfaltovenator vialidadi, Condorraptor currumili and Piatnitzkysaurus floresi) have been found in Toarcian-aged localities in the Cañadón Asfalto Formation (Chubut Province, Argentina) and, as a result, this unit has the highest known diversity of Jurassic theropods on the continent. A dentary with teeth in situ (MPEF-PV 6775) was found close to the base of this formation and differs from the other species from the unit with overlapping morphology. MPEF-PV 6775 resembles the dentary of the Late Jurassic North American theropod Ceratosaurus in being a robust bone, with a strong dorsal curvature in lateral view, and having relatively tall tooth crowns. MPEF-PV 6775 was scored in two matrices that have a broad taxonomic sample focused on Theropoda and the new specimen was recovered in large polytomies with other early neotheropods. A third analysis was conducted scoring MPEF-PV 6775 into a matrix focused in ceratosaurs because of its morphological similarities to Ceratosaurus. In this analysis, MPEF-PV 6775 was recovered as the sister taxon to Ceratosaurus, sharing with the later a ventral margin of the dentary that is strongly convex anteroposteriorly. These results suggest that MPEF-PV 6775 is a ceratosaurian and probably a ceratosaurid. Thus, our results indicate a higher taxonomic diversity of theropods and, in particular, ceratosaurs in the Cañadón Asfalto Formation. MPEF-PV 6775 seems to provide more evidence for the presence of ceratosaurids in the Jurassic of South America, together with isolated teeth described from the Late Jurassic of Uruguay and some analyses that recovered Eoabelisaurus and Berberosaurus within this clade.

DEVELOPMENT AND VALIDATION OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) FOR DETECTION OF LEPTOSPIR GENETIC MATERIAL

Primer systems were designed and tested and the method of loop-mediated isothermal amplification was validated for rapid diagnosis of leptospirosis. In general, five sets of primers were calculated and analyzed in situ and in vitro, which were ordered for synthesis and chosen for the future as the most promising. Indicators of analytical sensitivity and detection limits of the method were established, the specificity of the method in its tests with homologous and close to taxonomic characteristics heterologous samples of genetic material was proved and completed reproducibility of the method was established under the conditions of its staging on various laboratory equipment and with reagents from different manufacturers. The developed technique of loop-mediated amplification for the diagnosis of leptospirosis was tested on field samples and its diagnostic effectiveness was established.