Analyzing morphometry among extant and extinct species: A case study of genus Agassizia (Spatangoida: Echinoidea) / Análisis de la morfometría entre especies existentes y extintas: Un estudio de caso del género Agassizia (Spatangoida: Echinoidea)

Abstract Introduction: The genus Agassizia in Mexico is represented both in the fossil record by the species Agassizia regia† during the Miocene of Chiapas and by the extant species Agassizia excentrica on the Atlantic coast and Agassizia scrobiculata on the Pacific coast. Qualitative diagnosis and descriptions make it hard to distinguish morphological boundaries between species, especially in groups with fossils and recent representatives, increasing the level of complexity by having samples of disparate qualities and quantities. Objective: We propose the use of little explored statistical methods in the comparison of paleontological and biological populations. This methodology allowed us to resolve issues of missing values in a morphometric data set for the genus Agassizia. Methods: Using samples recently collected and specimens already housed in collections, we explore a routine of recovery of missing data MICE and the numerical and graphic analyses PERMANOVA, PCA, and SIMPER to compare morphometric parameters between these species for recognizing diagnostic characters. Results: Our results show a morphological difference in the length of the ambulacrum II and the length and width of the periproct and peristome structures, these being greater in A. scrobiculata, with a consistent pattern in both population samples not previously described. Conclusions: Quantitative morphometric comparisons can be an assertive and complementary tool to determine distinctive differentiation characteristics in species of the same genus. Comparative morphology reviews should be an ongoing exercise to keep taxonomic knowledge on both extinct and extant species up to date. Our research encourage the scientific community studying fossil populations to utilize quantitative and multivariate methods to strengthen their investigations.

Resumen Introducción: El género Agassizia en México está representado tanto en el registro fósil por la especie Agassizia regia† del Mioceno de Chiapas, como por las especies actuales Agassizia excentrica de la costa del Atlántico y Agassizia scrobiculata de la costa del Pacífico. Las descripciones y diagnosis cualitativas dificultan reconocer los limites morfológicos entre especies, especialmente en grupos con representantes fósiles y recientes, e incrementando el nivel de complejidad al tener muestras de cantidad y calidad desiguales. Objetivo: Proponemos el uso de métodos estadísticos poco explorados en la comparación de poblaciones paleontológicas y biológicas. Esta metodología nos permitió resolver problemas de valores faltantes en un conjunto de datos morfométricos para el género Agassizia. Métodos: Usando muestras recolectadas para este fin, así como provenientes de colecciones científicas, exploramos una rutina de recuperación de datos faltantes MICE, y los análisis numéricos y gráficos PERMANOVA, PCA y SIMPER para comparar parámetros morfométricos entre estas especies y reconocer caracteres de diagnóstico. Además, comparamos cuidadosamente los caracteres morfológicos descritos previamente en la literatura taxonómica y la descripción ambiental del hábitat actual de A. scrobiculata. Resultados: Nuestros resultados muestran una diferencia morfológica en la longitud del ambulacrum II y la longitud y anchura de las estructuras del periprocto y peristoma, siendo estas mayores en A. scrobiculata, con un patrón consistente en ambas muestras poblacionales no descrito previamente. El hábitat actual de las muestras de A. scrobiculata en la costa del Pacífico es un sistema costero poco profundo con sedimentos arenosos y temperaturas tropicales. Bahía Chamela comparte varias similitudes con la fauna y las condiciones ambientales previamente descritas en el Mioceno de Chiapas. Conclusiones: Las comparaciones morfométricas cuantitativas pueden ser una herramienta poderosa y complementaria para determinar caracteres distintivos de diferenciación en especies del mismo género. Las revisiones de morfología comparativa deben ser un ejercicio continuo para mantener actualizado el conocimiento taxonómico sobre las especies existentes y extintas. Nuestro trabajo busca incentivar a la comunidad científica que trabaja con poblaciones fósiles a explorar estos y otros métodos cuantitativos y multivariados para fortalecer sus investigaciones.

Phylogeography, colouration, and cryptic speciation across the Indo-Pacific in the sea urchin genus Echinothrix.

The sea urchins Echinothrix calamaris and Echinothrix diadema have sympatric distributions throughout the Indo-Pacific. Diverse colour variation is reported in both species. To reconstruct the phylogeny of the genus and assess gene flow across the Indo-Pacific we sequenced mitochondrial 16S rDNA, ATPase-6, and ATPase-8, and nuclear 28S rDNA and the Calpain-7 intron. Our analyses revealed that E. diadema formed a single trans-Indo-Pacific clade, but E. calamaris contained three discrete clades. One clade was endemic to the Red Sea and the Gulf of Oman. A second clade occurred from Malaysia in the West to Moorea in the East. A third clade of E. calamaris was distributed across the entire Indo-Pacific biogeographic region. A fossil calibrated phylogeny revealed that the ancestor of E. diadema diverged from the ancestor of E. calamaris ~ 16.8 million years ago (Ma), and that the ancestor of the trans-Indo-Pacific clade and Red Sea and Gulf of Oman clade split from the western and central Pacific clade ~ 9.8 Ma. Time since divergence and genetic distances suggested species level differentiation among clades of E. calamaris. Colour variation was extensive in E. calamaris, but not clade or locality specific. There was little colour polymorphism in E. diadema.

Cnidarian-bilaterian comparison reveals the ancestral regulatory logic of the ß-catenin dependent axial patterning.

In animals, body axis patterning is based on the concentration-dependent interpretation of graded morphogen signals, which enables correct positioning of the anatomical structures. The most ancient axis patterning system acting across animal phyla relies on ß-catenin signaling, which directs gastrulation, and patterns the main body axis. However, within Bilateria, the patterning logic varies significantly between protostomes and deuterostomes. To deduce the ancestral principles of ß-catenin-dependent axial patterning, we investigate the oral-aboral axis patterning in the sea anemone Nematostella-a member of the bilaterian sister group Cnidaria. Here we elucidate the regulatory logic by which more orally expressed ß-catenin targets repress more aborally expressed ß-catenin targets, and progressively restrict the initially global, maternally provided aboral identity. Similar regulatory logic of ß-catenin-dependent patterning in Nematostella and deuterostomes suggests a common evolutionary origin of these processes and the equivalence of the cnidarian oral-aboral and the bilaterian posterior-anterior body axes.

A new species and comparative morphology of Philippine sea biscuits (Echinoidea: Clypeaster/).

A new species of clypeasterid sea biscuit, Clypeaster brigitteae n. sp., is described from material collected in the Philippines at depths between 100 and 200 m. The new taxon increases the number of Clypeaster species recorded from the Philippines to nine, representing nearly a quarter of the world's diversity of the genus. Other Philippine species include: C. annandalei Koehler, 1922; C. fervens Koehler, 1922; C. humilis (Leske, 1778); C. japonicus Döderlein, 1885; C. latissimus (Lamarck, 1816); C. pateriformis Mortensen, 1948; C. reticulatus (Linnaeus, 1758); and C. virescens Döderlein, 1885. Using type material where available, each of these species is compared and contrasted with C. brigitteae n. sp. in tables consisting of new data derived from general test shape and size, petal structure, food grooves, plate architecture, internal structure, and morphology of spines, pedicellariae, and tube feet.

Efecto de la temperatura sobre la morfología y reproducción en el erizo de mar Arbacia dufresnii (Echinodermata: Echinoidea) / Effect of temperature on morphology and reproduction of sea urchin, Arbacia dufresnii (Echinodermata: Echinoidea)

Introducción: El aumento de los valores del CO2 atmosférico y el consecuente calentamiento del agua de mar constituyen factores de estrés para los ecosistemas marinos, renovando el interés en la fisiología de los equinoideos. El equinoideo Arbacia dufresnii es una especie ampliamente estudiada y con amplia distribución en los golfos norpatagónicos. Objetivo: Evaluar el efecto de temperatura del agua de mar sobre su morfología y reproducción. Métodos: Individuos de A. dufresnii fueron expuestos durante 10 semanas (Abril-Junio 2016) a tres tratamientos de temperatura (13 °C control, 15 °C y 17 °C) de acuerdo con las condiciones actuales del agua de mar y los valores esperados para el próximo siglo, según diferentes escenarios estimados. Al finalizar el experimento, diámetro y peso del individuo así como los pesos de los órganos internos fueron estadística e histológicamente analizados. Resultados: Se observó una mayor mortalidad en los individuos expuestos a las mayores temperaturas lo que podría deberse a un estrés en el individuo. Para el tratamiento control y el de 17 °C no se observaron cambios en los pesos secos de gónadas e intestino. En cambio, los individuos expuestos a 15 °C presentaron un aumento en el diámetro del caparazón y en el peso seco de gónadas e intestino. Los estadios gonadales aesta temperatura (15 °C) para machos fueron similares al ambiente a diferencia de las hembras que se observaron estadios más avanzados. Conclusiones: Esto indicaría que 15 °C podría ser la temperatura óptima para la asimilación del alimento y obtención de energía. A esta temperatura se observan cambios a corto plazo en los órganos de metabolismo rápido pero no en los órganos calcificados (caparazón y linterna de Aristóteles).

Introduction: The increase in atmospheric CO2 values and the consequent warming of seawater constitute stressors for marine ecosystems, renovating interest in echinoid physiology. The echinoid Arbacia dufresnii is a widely studied species with a wide distribution in the North Patagonian gulfs. Objective: Evaluate the effect of seawater temperature on its morphology and reproduction. Methods: Individuals were exposed for 10 weeks (April-June 2016) to three temperature treatments (13 °C control, 15 °C and 17 °C) according to the current conditions of seawater and the expected values for the next century. At the end of the experiment, diameter and weight of the whole individual and of its internal organs were statistically and histologically analyzed. Results: Individuals exposed to higher temperatures died in greater numbers. The control, and the 17 °C treatment, had no changes in dry gonad and intestine weights. However, the 15 °C group had wider diameter of test and a higher dry weight of gonads and intestine. At this temperature, the gonadal stages for males were similar to the environment, but females had more advanced stages. Conclusions: Temperature of 15°C could be optimal for assimilating food and obtaining energy. Gonads and intestine would be showing changes in the short term, unlike the calcified organs (test and Aristotle's lantern).

The influence of density on survival and larval development in the sea urchin Arbacia dufresnii (Echinodermata: Echinoidea)

Introduction: Density is one of the critical factors in echinoderm larvae for aquaculture purposes. Echinoplutei larvae are very sensitive to overcrowding. High culture density can lead to problems with bacteria or protozoa, decreasing survival and generating abnormal morphotypes. Objective: To evaluate the effect of culture density on survival and larval growth in the sea urchin Arbacia dufresnii. Methods: Two days after fertilization of A. dufresnii we we kept treatments at 1, 3, 5 and 10 larvae.ml-1, with three replicates each. We recorded survival and abnormal morphotypes periodically, as well as growth:somatic rod length, total length, and length of the post oral arms,. we applied generalized linear models. Results: Survival is dependent on density, time and replicates, and their interactions. Larval growth depended on density and time, also with interaction between the variables. The treatment of 5 larvae.ml-1 had the highest survival and larval condition. Conclusions: Larval culture of A. dufresnii had the best results at 5 larvae.ml-1.

Introducción: La densidad es uno de los factores críticos para las larvas de equinodermo para fines de acuicultura. Las larvas de Echinoplutei son muy sensibles al hacinamiento. Una alta densidad de cultivo puede provocar problemas con bacterias o protozoos, disminuyendo la supervivencia y generando morfotipos anormales. Objetivo: Evaluar el efecto de la densidad de cultivo sobre la supervivencia y el crecimiento larvario del erizo de mar Arbacia dufresnii. Métodos: Dos días después de la fertilización de A. dufresnii se mantuvieron los tratamientos a 1, 3, 5 y 10 larvas.ml-1, con tres repeticiones cada uno. Registramos la supervivencia y los morfotipos anormales periódicamente, así como el crecimiento: longitud de la varilla somática, longitud total y longitud de los brazos posorales. aplicamos modelos lineales generalizados. Resultados: La supervivencia depende de la densidad, el tiempo y las réplicas y sus interacciones. El crecimiento larvario dependió de la densidad y el tiempo, también de la interacción entre las variables. El tratamiento de 5 larvas.ml-1 tuvo la mayor supervivencia y condición larvaria. Conclusiones: El cultivo larvario de A. dufresnii tuvo los mejores resultados con 5 larvas.ml-1.

Diversity and biostratigraphy of the late Oligocene-late Miocene sand dollars (Echinoidea: Scutelliformes) of Argentina and Uruguay / Diversidad y bioestratigrafía de las galletas de mar (Echinoidea: Scutelloida: Scutellifomes) del Oligoceno tardío-Mioceno tardío de Argentina y Uruguay

Introduction: Scutelliforms were diverse and widespread in shallow marine environments during Neogene times in South America. Nevertheless, they have almost never been used as biostratigraphic tools. Objective: To provide a refined stratigraphic frame useful for calibrating temporal dimensions of scutelliform diversity from Argentina and Uruguay and its correlation with the molluscan assemblages previously proposed. Methods: A detailed survey of their geographic and stratigraphic provenance was carried out. We revised both the bibliography and collections (institutional and from our own field work). Results: The group is represented by 14 species belonging to six genera, and four assemblages were identified. Numerical dates of the Neogene marine rocks obtained recently allowed their placement in a chronological scheme: "Iheringiella" sp. A is restricted to the late Oligocene, the genera Camachoaster and "Eoscutella" and the species Monophoraster telfordi to the early Miocene, Abertella gualichensis and Abertella miskellyi to the middle Miocene, and Monophoraster duboisi, Amplaster coloniensis and Amplaster ellipticus to the late Miocene. Non-lunulate scutelliforms are not restricted to the late Oligocene as previously supposed. The oldest occurrence of the genus Monophoraster corresponds to the early Miocene, and along with Iheringiella are long-living taxa that embrace the 25.3 Ma-18.1 Ma (Iheringiella patagonensis) and approximately 15 Ma-6.48 Ma (Monophoraster darwini) intervals. The presence of Iheringiella in the early Miocene of northeastern Patagonia is corroborated, reaching there its northernmost distribution. Monophoraster darwini has a temporal range from the late Miocene (where it was previously thought to be restricted) back to the middle Miocene, since this is the species yielded in the well-known and discussed "Monophoraster and Venericor Beds". Conclusions: The Paleogene-Neogene scutelliforms of Argentina and Uruguay range from the late Oligocene to the late Miocene. There is a good correspondence among the numerical ages, molluscan biozones and scutelliform assemblages.

Introducción: Realizamos una síntesis actualizada de la composición taxonómica de las asociaciones de escutélidos del Oligoceno tardío-Mioceno tardío de Argentina y Uruguay. Objetivo: Ubicar los escutélidos en un marco estratigráfico refinado, a los efectos de observar su diversidad a lo largo del tiempo y la correlación de sus ensambles con los ya conocidos de moluscos. Métodos: Revisamos en detalle la procedencia geográfica y estratigráfica de los ejemplares y su asignación taxonómica, basándonos tanto en la bibliografía como en colecciones institucionales y producto de nuestras campañas de colecta. Resultados: El grupo está representado por 14 especies agrupadas en seis géneros, siendo identificadas en cuatro ensambles. Las edades numéricas recientemente obtenidas permitieron su ubicación en un esquema cronológico: "Iheringiella" sp. A se restringe al Oligoceno tardío, Camachoaster y "Eoscutella", y Monophoraster telfordi al Mioceno temprano, Abertella gualichensis y Abertella miskellyi al Mioceno medio, y Monophoraster duboisi, Amplaster coloniensis y Amplaster ellipticus al Mioceno tardío. Los escutélidos sin lúnula no están restringidos al Oligoceno tardío como se suponía. Monophoraster aparece en el Mioceno temprano y junto con Iheringiella abarcan largos períodos de tiempo, encontrándoselos entre los 25.3 Ma-18.1 Ma (Iheringiella patagonensis) y aproximadamente los 15 Ma-6.48 Ma (Monophoraster darwini). Se corrobora la presencia de Iheringiella en el Mioceno temprano del noreste de Patagonia, donde alcanza su punto más septentrional. Monophoraster darwini se encuentra desde el Mioceno medio hasta el Mioceno tardío (lapso para el cual previamente se consideraba restringido), ya que se trata de la especie presente en los bien conocidos "estratos con Monophoraster y Venericor". Conclusiones: Los escutélidos del Paleógeno-Neógeno de Argentina y Uruguay se encuentran desde el Oligoceno tardío hasta el Mioceno tardío. Hay una buena correspondencia entre las edades numéricas, las biozonas de moluscos y los ensambles de escutélidos.

Gross and microscopic pathology of West Indian sea eggs (Tripneustes ventricosus).

In this study, we performed comprehensive pathology examinations on 83 Tripneustes ventricosus from 11 locations on St. Kitts to build baseline data necessary for disease diagnosis in this species. Gross abnormalities were observed in 23/83 (28%) urchins and included spine loss, visceral hyperpigmentation, test discoloration, and test ulceration. Ciliates were the only protists identified in this study via examination of tissue wet mounts and histology, documented in 50/83 (60%) urchins. Microscopic observations associated with visibly abnormal status included muscle necrosis, test and appendage inflammation, appendage (tube feet, spines, and pedicellariae) degeneration, severe coelomocytosis, and generalized hypermelanosis. Enterocyte intranuclear inclusion bodies, microbial aggregates, nerve pigmentation, enteric pigmentation, integument-associated crustaceans, and encysted metazoan parasites were of uncertain pathological significance. The etiology for any lesion was not microscopically apparent, contrasting literature implicating common marine bacteria in urchin diseases. This study highlights the importance of histopathology in urchin disease investigations and facilitates the recognition of disease in T. ventricosus.

Effect of dopamine on early larvae of sea urchins, Mesocentrotus nudus and Strongylocentrotus intermedius.

Larvae of many echinoids are known to be phenotypically plastic and capable of changing the growth rate of their post-oral arms depending on the microalgae concentration in their habitat. As literature data show, developing larvae use chemosensation to detect algae in the environment and "adjust" the rate of growth of their post-oral arms through dopamine signaling. According to our results, dopamine has a significant effect on the post-oral arm growth in early larvae of two sea urchin species, Mesocentrotus nudus and Strongylocentrotus intermedius. The dopamine effect depends on concentration: the higher the dopamine concentration in the water, the shorter the post-oral arms. We suggest that the pattern of response to variation in dopamine concentration, manifested by early larvae of both species, is similar to that observed at different concentrations of microalgae.

Quantitative 3D structural analysis of the cellular microstructure of sea urchin spines (II): Large-volume structural analysis.

Biological cellular materials have been a valuable source of inspiration for the design of lightweight engineering structures. In this process, a quantitative understanding of the biological cellular materials from the individual branch and node level to the global network level in 3D is required. Here we adopt a multiscale cellular network analysis workflow demonstrated in the first paper of this work series to analyze the biomineralized porous structure of sea urchin spines from the species Heterocentrotus mamillatus over a large volume (ca. 0.32mm3). A comprehensive set of structural descriptors is utilized to quantitatively delineate the long-range microstructural variation from the spine center to the edge region. Our analysis shows that the branches gradually elongate (~50% increase) and thicken (~100% increase) from the spine center to edge, which dictates the spatial variation of relative density (from ~12% to ~40%). The branch morphology and network organization patterns also vary gradually with their positions and orientations. Additionally, the analysis of the cellular network of individual septa provides the interconnection characteristics between adjacent septa, which are the primary structural motifs used for the construction of the cellular structure in the edge region. Lastly, combining the extracted long-range cellular network and finite element simulations allows us to efficiently examine the spatial and orientational dependence of local effective Young's modulus across the spine's radius. The structural-mechanical analysis here sheds light on the structural designs of H. mamillatus' porous spines, which could provide important insights for the design and modeling of lightweight yet strong and damage-tolerant cellular materials. STATEMENT OF SIGNIFICANCE: Previous investigations on the cellular structures of sea urchin spines have been mainly based on 2D measurements or 3D quantification of small volumes with limited structural parameters. This limits our understanding of the interplay between the 3D microstructural variations and the mechanical properties in sea urchin spines, which hence constrains the derivation of the underlying principles for bio-inspired designs. This work utilizes our multiscale 3D network analysis, for the first time, to quantify the 3D cellular network and its variation across large volumes in sea urchin spines from individual branch and node level to the cellular network level. The network analysis demonstrated here is expected to be of great interest to the fields of biomineralization, functional biological materials, and bio-inspired material design.

Quantitative 3D structural analysis of the cellular microstructure of sea urchin spines (I): Methodology.

The mineralized skeletons of echinoderms are characterized by their complex, open-cell porous microstructure (also known as stereom), which exhibits vast variations in pore sizes, branch morphology, and three-dimensional (3D) organization patterns among different species. Quantitative description and analysis of these cellular structures in 3D are needed in order to understand their mechanical properties and underlying design strategies. In this paper series, we present a framework for analyzing such structures based on high-resolution 3D tomography data and utilize this framework to investigate the structural designs of stereom by using the spines from the sea urchin Heterocentrotus mamillatus as a model system. The first paper here reports the proposed cellular network analysis framework, which consists of five major steps: synchrotron-based tomography and hierarchical convolutional neural network-based reconstruction, machine learning-based segmentation, cellular network registration, feature extraction, and data representation and analysis. This framework enables the characterization of the porous stereom structures at the individual node and branch level (~10 µm), the local cellular level (~100 µm), and the global network level (~1 mm). We define and quantify multiple structural descriptors at each level, such as node connectivity, branch length and orientation, branch profile, ring structure, etc., which allows us to investigate the cellular network construction of H. mamillatus spines quantitatively. The methodology reported here could be tailored to analyze other natural or engineering open-cell porous materials for a comprehensive multiscale network representation and mechanical analysis. STATEMENT OF SIGNIFICANCE: The mechanical robustness of the biomineralized porous structures in sea urchin spines has long been recognized. However, quantitative cellular network representation and analysis of this class of natural cellular solids are still limited in the literature. This constrains our capability to fully understand the mechanical properties and design strategies in sea urchin spines and other similar echinoderms' porous skeletal structures. Combining high-resolution tomography and computer vision-based analysis, this work presents a multiscale 3D network analysis framework, which allows for extraction, registration, and quantification of sea urchin spines' complex porous structure from the individual branch and node level to the global network level. This 3D structural analysis is relevant to a diversity of research fields, such as biomineralization, skeletal biology, biomimetics, material science, etc.

Use of sea urchin spines with chitosan gel for biodegradable film production.

In recent years, the film production from natural polymers has considerably increased in food industry as an alternative to the petroleum based synthetic films. Chitosan is one of the most preferred biopolymers for bio-based film production, due to its biocompatibility, biodegradability, antioxidant activity and antimicrobial properties. Because of its hydrophilic properties, chitosan based films dissolve in water, limiting its uses in industry, to overcome this problem; we mixed 200 and 400 mg of the sea urchin spine powder (SUSP) with 20 mL chitosan gel respectively, to obtain a hydrophobic film. The chitosan films prepared with 200 mg SUSP showed a rise in the degree of contact angle from 70.2° to 107° providing hydrophobicity properties. On the other hand, addition of 400 mg of SUSP to chitosan film resulted in a contact angle of 96.1°. Moreover, the antioxidant activity and thermal stability of the films were increased in the presence of SUSP. Fourier Transform Infrared Spectrophotometry results proved the interactions between chitosan and SUSP. Chitosan films have smooth surface while SUSP blended films have rough surface morphology. These results demonstrated that SUSP is needed to improve the properties of chitosan films for usage in food industry.

Cell rearrangement induced by filopodial tension accounts for the late phase of convergent extension in the sea urchin archenteron.

George Oster was a pioneer in using mechanical models to interrogate morphogenesis in animal embryos. Convergent extension is a particularly important morphogenetic process to which George Oster gave significant attention. Late elongation of the sea urchin archenteron is a classic example of convergent extension in a monolayered tube, which has been proposed to be driven by extrinsic axial tension due to the activity of secondary mesenchyme cells. Using a vertex-based mechanical model, we show that key features of archenteron elongation can be accounted for by passive cell rearrangement due to applied tension. The model mimics the cell elongation and the Poisson effect (necking) that occur in actual archenterons. We also show that, as predicted by the model, ablation of secondary mesenchyme cells late in archenteron elongation does not result in extensive elastic recoil. Moreover, blocking the addition of cells to the base of the archenteron late in archenteron elongation leads to excessive cell rearrangement consistent with tension-induced rearrangement of a smaller cohort of cells. Our mechanical simulation suggests that responsive rearrangement can account for key features of archenteron elongation and provides a useful starting point for designing future experiments to examine the mechanical properties of the archenteron.

A new ophiocistioid with soft-tissue preservation from the Silurian Herefordshire Lagerstätte, and the evolution of the holothurian body plan.

Reconstructing the evolutionary assembly of animal body plans is challenging when there are large morphological gaps between extant sister taxa, as in the case of echinozoans (echinoids and holothurians). However, the inclusion of extinct taxa can help bridge these gaps. Here we describe a new species of echinozoan, Sollasina cthulhu, from the Silurian Herefordshire Lagerstätte, UK. Sollasina cthulhu belongs to the ophiocistioids, an extinct group that shares characters with both echinoids and holothurians. Using physical-optical tomography and computer reconstruction, we visualize the internal anatomy of S. cthulhu in three dimensions, revealing inner soft tissues that we interpret as the ring canal, a key part of the water vascular system that was previously unknown in fossil echinozoans. Phylogenetic analyses strongly suggest that Sollasina and other ophiocistioids represent a paraphyletic group of stem holothurians, as previously hypothesized. This allows us to reconstruct the stepwise reduction of the skeleton during the assembly of the holothurian body plan, which may have been controlled by changes in the expression of biomineralization genes.

A phylogenomic resolution of the sea urchin tree of life.

BACKGROUND: Echinoidea is a clade of marine animals including sea urchins, heart urchins, sand dollars and sea biscuits. Found in benthic habitats across all latitudes, echinoids are key components of marine communities such as coral reefs and kelp forests. A little over 1000 species inhabit the oceans today, a diversity that traces its roots back at least to the Permian. Although much effort has been devoted to elucidating the echinoid tree of life using a variety of morphological data, molecular attempts have relied on only a handful of genes. Both of these approaches have had limited success at resolving the deepest nodes of the tree, and their disagreement over the positions of a number of clades remains unresolved. RESULTS: We performed de novo sequencing and assembly of 17 transcriptomes to complement available genomic resources of sea urchins and produce the first phylogenomic analysis of the clade. Multiple methods of probabilistic inference recovered identical topologies, with virtually all nodes showing maximum support. In contrast, the coalescent-based method ASTRAL-II resolved one node differently, a result apparently driven by gene tree error induced by evolutionary rate heterogeneity. Regardless of the method employed, our phylogenetic structure deviates from the currently accepted classification of echinoids, with neither Acroechinoidea (all euechinoids except echinothurioids), nor Clypeasteroida (sand dollars and sea biscuits) being monophyletic as currently defined. We show that phylogenetic signal for novel resolutions of these lineages is strong and distributed throughout the genome, and fail to recover systematic biases as drivers of our results. CONCLUSIONS: Our investigation substantially augments the molecular resources available for sea urchins, providing the first transcriptomes for many of its main lineages. Using this expanded genomic dataset, we resolve the position of several clades in agreement with early molecular analyses but in disagreement with morphological data. Our efforts settle multiple phylogenetic uncertainties, including the position of the enigmatic deep-sea echinothurioids and the identity of the sister clade to sand dollars. We offer a detailed assessment of evolutionary scenarios that could reconcile our findings with morphological evidence, opening up new lines of research into the development and evolutionary history of this ancient clade.

Structural design of the echinoid's trabecular system.

The multi-plated skeleton of echinoids is made of the stereom, a light-weight construction which resembles a micro-beam framework. Although the two-dimensional design of the stereom has been studied, its spatial architecture is only little known. It is, however, imperative to understand the spatial architecture of the trabecular system in order to interpret its structural principles of this load-bearing construction. The echinoid's trabecular system is thus analyzed in-depth with respect to eight topological descriptors. The echinoid's plates are divided into two regions, the center of which consists of an unordered stereom, and the margin which consists of an ordered stereom. The eight trabecular descriptors indicate that the basal topology of the two plate regions are similar. The trabecular system predominantly consists of short and stocky trabeculae that show little tortuosity. The majority of trabeculae intersect in a 3N configuration, where three trabeculae intersect in one common node. Trabeculae in the 3N configuration intersect in an angle of around 120° resulting in a planar and triangular motif. These planar elements, when arranged in an angular off-set, can resist multi-dimensional loads. Results also show that the trabecular orientation perpendicular to the plate's surface is at an angle of 60°. The trabecular orientation in the plate's horizontal plane is directional. Both trabecular orientations reflect a construction which is capable of resisting applied loads and can distribute these loads over the entire skeleton. The spatial architecture of the echinoid's trabecular system is thus considered to be a performative light-weight and load-bearing system.

Structural stress response of segmented natural shells: a numerical case study on the clypeasteroid echinoid Echinocyamus pusillus.

The skeleton of Echinocyamus pusillus is considered as an exceptional model organism for structural strength and skeletal integrity within the echinoids as demonstrated by the absence of supportive collagenous fibres between single plates and the high preservation potential of their skeletons. The structural principles behind this remarkably stable, multi-plated, light-weight construction remain hardly explored. In this study, high-resolution X-ray micro-computed tomography, finite-element analysis and physical crushing tests are used to examine the structural mechanisms of this echinoid's skeleton. The virtual model of E. pusillus shows that the material is heterogeneously distributed with high material accumulations in the internal buttress system and at the plate boundaries. Finite-element analysis indicates that the heterogeneous material distribution has no effect on the skeleton's strength. This numerical approach also demonstrates that the internal buttress system is of high significance for the overall skeletal stability of this flattened echinoid. Results of the finite-element analyses with respect to the buttress importance were evaluated by physical crushing tests. These uniaxial compression experiments support the results of the simulation analysis. Additionally, the crushing tests demonstrate that organic tissues do not significantly contribute to the skeletal stability. The strength of the echinoid shell, hence, predominantly relies on the structural design.

Synthesis of Zinc Oxide Nanorods via the Formation of Sea Urchin Structures and Their Photoluminescence after Heat Treatment.

A protocol for the aqueous synthesis of ca. 1-µm-long zinc oxide (ZnO) nanorods and their growth at intermediate reaction progression is presented, together with photoluminescence (PL) characteristics after heat treatment at temperatures of up to 1000 °C. The existence of solitary rods after the complete reaction (60 min) was traced back to the development of sea urchin structures during the first 5 s of the precipitation. The rods primarily formed in later stages during the reaction due to fracture, which was supported by the frequently observed broken rod ends with sharp edges in the final material, in addition to tapered uniform rod ends consistent with their natural growth direction. The more dominant rod growth in the c direction (extending the length of the rods), together with the appearance of faceted surfaces on the sides of the rods, occurred at longer reaction times (>5 min) and generated zinc-terminated particles that were more resistant to alkaline dissolution. A heat treatment for 1 h at 600 or 800 °C resulted in a smoothing of the rod surfaces, and PL measurements displayed a decreased defect emission at ca. 600 nm, which was related to the disappearance of lattice imperfections formed during the synthesis. A heat treatment at 1000 °C resulted in significant crystal growth reflected as an increase in luminescence at shorter wavelengths (ca. 510 nm). Electron microscopy revealed that the faceted rod structure was lost for ZnO rods exposed to temperatures above 600 °C, whereas even higher temperatures resulted in particle sintering and/or mass redistribution along the initially long and slender ZnO rods. The synthesized ZnO rods were a more stable Wurtzite crystal structure than previously reported ball-shaped ZnO consisting of merging sheets, which was supported by the shifts in PL spectra occurring at ca. 200 °C higher annealing temperature, in combination with a smaller thermogravimetric mass loss occurring upon heating the rods to 800 °C.

Cryptic speciation in pan-tropical sea urchins: a case study of an edge-of-range population of Tripneustes from the Kermadec Islands.

Tripneustes is one of the most abundant and ecologically significant tropical echinoids. Highly valued for its gonads, wild populations of Tripneustes are commercially exploited and cultivated stocks are a prime target for the fisheries and aquaculture industry. Here we examine Tripneustes from the Kermadec Islands, a remote chain of volcanic islands in the southwest Pacific Ocean that mark the boundary of the genus' range, by combining morphological and genetic analyses, using two mitochondrial (COI and the Control Region), and one nuclear (bindin) marker. We show that Kermadec Tripneustes is a new species of Tripneustes. We provide a full description of this species and present an updated phylogeny of the genus. This new species, Tripneustes kermadecensis n. sp., is characterized by having ambulacral primary tubercles occurring on every fourth plate ambitally, flattened test with large peristome, one to two occluded plates for every four ambulacral plates, and complete primary series of interambulacral tubercles from peristome to apex. It appears to have split early from the main Tripneustes stock, predating even the split of the Atlantic Tripneustes lineage. Its distinction from the common T. gratilla and potential vulnerability as an isolated endemic species calls for special attention in terms of conservation.

Differences in Larval Arm Movements Correlate with the Complexity of Musculature in Two Phylogenetically Distant Echinoids, Eucidaris tribuloides (Cidaroidea) and Lytechinus variegatus (Euechinoidea).

Within a common body plan, echinoid planktotrophic larvae are morphologically diverse, with variations in overall size, the length, and number of arms and the presence or absence of epidermal structures. In this report, we are interested in variation in larval arm-flexing behavior and correlated differences in larval musculature. Larvae of the cidaroid Eucidaris tribuloides exhibit conspicuous and regular arm-flexing behavior. In contrast, Lytechinus variegatus, a representative of the euechinoid clade, does not exhibit this behavior. We hypothesized that there were differences in musculature that correlated with this behavioral contrast and compared the development and structure of larval muscles between these species. We report substantial differences in some aspects of larval musculature. In addition to previously described oral musculature, both larvae possessed polygon-shaped musculature at the basal end of the larva. However, larval musculature in E. tribuloides was larger and contained additional muscles not observed in larvae of L. variegatus. Therefore, a conspicuous larval behavior consisting of repeated flexing of the postoral and posterodorsal larval arms was correlated with a larger, more complex musculature. This simple contrast indicates that larval musculature not associated with endoderm evolves in a manner that relates to differences in larval behavior and that additional comparisons are warranted.