Sea Urchin Research Articles

Does the righting behaviour effectively reflect stress in Arbacia dufresnii Arbacioida: Arbaciidae)?

Introduction: Righting behaviour has been used as a health indicator in response to stressor variables. Using this parameter in aquaculture could help to reduce mortality and improve welfare in the sea urchin Arbacia dufresnii culture.
 Objective: The purpose of this study was to determine the effect of sex, diameter, and three stressor factors on the righting behaviour of the sea urchin A. dufresnii.
 Methods: A total of 300 animals were evaluated for complete righting behaviour (CRB) time, with 100 of them also recording half righting behaviour (HRB) time. Three stressors were applied to the animals: serial repetitions (three successive turnings), temperature (24-hour shock), and spawning induction with KCl injection. A stopwatch was used to record the time, and a precision calliper was used to measure the diameter.
 Results: Righting time was discovered to be diameter dependent but sex independent. The upper temperature limit of 19 °C had a significant effect on righting behaviour compared to 16 °C and 13 °C with CRB times up to 150 seconds.  Serial repetitions and spawning had no significant effect. However, based on the recorded times, it can be deduced that spawning had an effect on the health of the animals, with CRB times of up to 150 seconds compared to the control, with lower times.
 Conclusions: Complete righting behaviour appears to be an optimal indicator for evaluating the health and condition of the sea urchin A. dufresnii, but more tests would be performed to confirm the effect of the control treatment on post-spawning stress.

Effect of gamete aging on fertilization success of the sea urchin Arbacia dufresnii (Arbacioida: Arbaciidae)

Introduction: Short-term gametes storage is an inexpensive and simple technique that allows the use of the same batch of eggs or sperm at different times, maximizing the application of research protocols and the use of gametes in production. Arbacia dufresnii is a sea urchin species with proven aquaculture potential and already used in the nutraceutical industry. Aging of its gametes is unknown and is a needed information to scale up the production.
 Objective: Determine the effect of male and female gamete aging on the fertilization success of Arbacia dufresnii. This will allow optimizing the use of gametes after collection decoupling spawning from fertilization.
 Methods: A. dufresnii individuals were induced to spawn and gametes were kept at 12 ± 1 °C throughout each bioassay. Sperm was separated into two treatments: activated sperm in seawater (AS), and dry sperm (DS). Two bioassays were made: Bioassay 1 evaluated the effect of time on fertility by performing fertilization tests at 0 h, 24 h, 48 h, 72 h, and 96 h after spawning. Bioassay 2 evaluated the contribution of each type of aged gamete on fertility, combining aged gametes (96 h) with fresh gametes (0 h).
 Results: Bioassay 1: the fertilization success obtained by combining eggs (E) with AS or DS presented important differences. While the fertilization success remained acceptable (greater than 50 %) for up to 72 h using ExDS, it only remained acceptable for up to 48 h using ExAS. Bioassay 2: acceptable fertilization success was found by combining aged E (96 h) with fresh sperm, or aged DS (96 h) with fresh E, but not using aged AS with fresh E.
 Conclusions: The findings of this work show that fertilization success in A. dufresnii gametes remains relatively unchanged for up to 48 h after spawning when combining ExAS, and for up to 72 h when combining ExDS. However, when combining aged E or aged DS with a fresh gamete, post-collection fertilization can be extended up to 96 h. In this work, the first steps have been taken to understand the conservation time of A. dufresnii gametes with minimum intervention.

Tropomyosin induces the synthesis of magnesian calcite in sea urchin spines

Calcium carbonate is present in many biominerals, including in the exoskeletons of crustaceans and shells of mollusks. High Mg-containing calcium carbonate was synthesized by high temperatures, high pressures or high molecular organic matter. For example, biogenic high Mg-containing calcite is synthesized under strictly controlled Mg concentration at ambient temperature and pressure. The spines of sea urchins consist of calcite, which contain a high percentage of magnesium. In this study, we investigated the factors that increase the magnesium content in calcite from the spines of the sea urchin, Heliocidaris crassispina. X-ray diffraction and inductively coupled plasma mass spectrometry analyses showed that sea urchin spines contain about 4.8% Mg. The organic matrix extracted from the H. crassispina spines induced the crystallization of amorphous phase and synthesis of magnesium-containing calcite, while amorphous was synthesized without SUE (sea urchin extract). In addition, aragonite was synthesized by SUE treated with protease-K. HC tropomyosin was specifically incorporated into Mg precipitates. Recombinant HC-tropomyosin induced calcite contained 0.1–2.5% Mg synthesis. Western blotting of sea urchin spine extracts confirmed that HC tropomyosin was present in the purple sea urchin spines at a protein weight ratio of 1.5%. These results show that HC tropomyosin is one factor that increases the magnesium concentration in the calcite of H. crassispina spines.

Temporal dynamics of bacterial populations in recirculating aquaculture systems for sea urchin production

Introduction: Sea urchin aquaculture is a rising industry, and in consequence, there is a need to establish optimal culture parameters to ensure the health of the cultured animals.
 Objective: To evaluate the bacterial counts in the seawater of sea urchin (Arbacia dufresnii) aquaculture recirculating systems (RAS).
 Methods: The bacteriological water quality of two RAS containing sea urchins was determined. For approximately two months, weekly water samples were taken. The bacteriological quality was determined by counting total aerobic heterotrophic populations, lactic acid bacteria, enterobacterias and genus Vibrio. Physicochemical parameters were also measured.
 Results: There was no presence of disease or mortality. Enterobacteria and lactic acid bacteria were not detected from both RAS systems. The number of animals had an important effect on the observed difference in the count of total bacteria and Vibrio spp. In RAS 1 the maximum counts of total bacteria and Vibrio spp. were 2.8 x 105 ± 1.7 x 105 and 1.45 x 105 ± 3.6 x 104 UFC ml-1, respectively. In RAS 2 total bacteria and Vibrio spp. exhibited repetitive behavior over time influenced in part by water exchange and mainly by feeding. The results indicate that periodic water changes ensure a limited growth of bacterial strains as Vibrio and other bacteria.
 Conclusions: Our results suggests that the bacterial count levels recorded in this study can be used as a threshold or safety limit for Arbacia dufresnii aquaculture.

Study of histopathology on Arbacia lixula (Arbaciidae: Arbacioida) and Paracentrotus lividus (Parachinidae: Camarodonta) with bald sea urchin disease symptoms in Gran Canaria Island, Spain

Introduction: Sea urchin diseases have been documented in several locations worldwide, with reported occurrences of bacterial, protozoan, fungal, and algal infections.
 Objective: This study aimed to investigate pathogen agents in populations of Arbacia lixula and Paracentrotus lividus along the coast of Gran Canaria Island (Central-East Atlantic, Spain).
 Methods: Sampling was conducted at San Cristobal beach, on the Northeast side of the island, where sea urchins were manually collected from depths of 1-3 m during June, July, and October 2022. Swab samples were taken from the external and internal areas of the lesions and cultured on various media plates.
 Results: Eight different pathogen agents, including bacteria and fungi, were identified, with Vibrio alginolyticus being the most frequently observed bacteria in all diseased sea urchin samples. Additionally, ciliated protozoans were found within the tests, potentially acting as opportunistic parasites.
 Conclusions: This research provides a unique perspective on bald sea urchin disease by identifying a significant number of associated pathogens, including Candida, previously unreported in diseased organisms. Furthermore, the study highlights the presence of an inflammatory response in tissues with bacterial colonies, offering crucial insights into understanding this sea urchin disease.

Variation of the diet of the sea urchin (Diadema mexicanum (Diadematoida: Diadematidae)) according to its size in the Eastern Tropical Pacific

Introduction: The sea urchin Diadema mexicanum, due to its bioerosion activity, is considered of ecological importance. This phenomenon could negatively or positively affect coral reef ecosystems. The bioerosion process varies according to the abundance and size of the sea urchin.
 Objective: Juvenile organisms possess different metabolic needs compared to adults, so knowing their stomach content according to size allows us to quantify the selection of substrate bioeroded.
 Methods: To determine this, D. mexicanum individuals were collected in 12 sites from January 2009 to September 2010 along the Eastern Tropical Pacific coast. The stomach content was categorized in Carbonated Fraction (CF), Non-Carbonated Fraction (NCF), and Organic Matter (OM). Stomach content was analyzed according to a) juvenile (< 2.5 cm) or adult (> 2.5 cm) stage and b) locality.
 Results: Juveniles presented the following stomach content average percentages: 20.7 % OM, 12 % NCF and 67.9 % CF; and adults: 11.4 % OM, 14.8 % NCF and 73.8 % CF. Based on a Wilcoxon test and a Kendall linear regression, the following results were obtained. The carbonated fraction in the stomach increased by 0.47 units on average for every cm of growth (p < 0.05). OM consumed by D. mexicanum increases only 0.05 units for every cm of growth (p < 0.05). We found a difference of stomach content depending on the site (p < 0.05) and life stage (p < 0.05). Localities like Huatulco and Coco presented significant differences that could be related to local oceanographic conditions.
 Conclusions: We relate these changes of the stomach fractions to the necessity of the juvenile sea urchins for nutrients to maintain their growth. The amount of OM is crucial for the development of early stages, meaning that there is a difference in substrate selection associated with growth.

Studies on Secondary Metabolites and In vitro and In silico Anticholinesterases Activities of the Sea Urchin Echinometra mathaei Crude Venoms from the Persian Gulf-Bushehr

Background: Echinoderms are a unique source of amazing secondary metabolites with a wide spectrum of biological activities. Several species of sea urchins contain various toxins and biologically active metabolites. One of the most attractive approaches to treat Alzheimer's disease is searching for effective marine natural products with cholinesterase inhibitory activities. Objective: The current study is designed to investigate the in vitro and in silico acetylcholinesterase and butyrylcholinesterase inhibitory activities of the Persian Gulf echinoderm sea urchin Echinometra mathaei venom and related chemical compounds. Methods: The experiments for LD50, total protein, protein bands, in vitro cholinesterase inhibitory activities, the identity of secondary metabolites, and the in silico evaluations, respectively, were performed by Spearman-Karber, Lowry, SDS-PAGE, Ellman's spectroscopic, GC-MS, and docking methods. Results: The LD50 (IV rat) of the spine, gonad, and coelomic fluid from sea urchin samples were 2.231 ± 0.09, 1.03 ± 0.05, and 1.12 ± 0.13 mg/ml, respectively. The SDS-PAGE and total protein studies showed that at least a portion of the venom is protein in nature. GC-MS analysis of the identified samples revealed 12, 23, and 21 compounds with different chemical types, including alkaloids, terpenes, and steroids, respectively. According to the results, all samples act as significant inhibitors of both enzymes. In silico data for the identified compounds also confirmed the experimental results. Conclusion: The alkaloid compound 6H-Indolo[3,2,1-de] [1,5] naphthyridine-6-one,1,2,3a,4,5- hexahydro-8-hydroxy-3-methyl (C7) had the highest affinity for both enzymes. Further research is needed to determine whether C7 could be a therapeutic candidate for Alzheimer's disease.

Bacteriological study of the sea urchin Paracentrotus lividus (Lamarck, 1816) gonads associated with seawater physico-chemistry and environmental factors in the Mediterranean Sea of Morocco

The preservation of marine ecosystems and the safety of harvested seafood have elicited global concern. In this study, we evaluated bacterial contamination levels in sea urchin Paracentrotus lividus gonads with relation to environmental factors and the physicochemical parameters of seawater. This investigation aimed to assess bacterial contamination levels of Total Aerobic Mesophilic Bacteria (TAMB), Enterobacteriaceae (ETB), Fecal Coliforms (FC), Staphylococcus aureus(STAPH), Escherichia coli (E. coli), and Salmonella spp. (SALM) in sea urchin gonads collected from the Tangier coast between December 2020 and November 2021, per Moroccan regulations (NM 08.7.040). Twenty-one physicochemical, environmental, and biological parameters were assessed in 360 samples of P. lividus from four seasons. The results of these parameters were processed using multivariate statistical approaches. It was found that almost all biological parameters had concentrations below Moroccan regulatory limits, except for FC in July (173 CFU/g). Furthermore, E. coli and SALM were absent. Principal component analysis (PCA) demonstrated that the F1 component was associated with a combination of environmental, microbiological, and anthropogenic factors. These associations contributed to 59.02% of all data variations and were strongly influenced by seasonality. The F2 component contributed to 26.33% of all data variations. These findings may aid decision-makers in the protection of public health in sensitive locations such as the Tangier coast in northern Morocco.

The positioning mechanics of microtubule asters in Drosophila embryo explants.

Microtubule asters are essential in localizing the action of microtubules in processes including mitosis and organelle positioning. In large cells, such as the one-cell sea urchin embryo, aster dynamics are dominated by hydrodynamic pulling forces. However, in systems with more densely positioned nuclei such as the early Drosophila embryo, which packs around 6000 nuclei within the syncytium in a crystalline-like order, it is unclear what processes dominate aster dynamics. Here, we take advantage of a cell cycle regulation Drosophila mutant to generate embryos with multiple asters, independent from nuclei. We use an ex vivo assay to further simplify this biological system to explore the forces generated by and between asters. Through live imaging, drug and optical perturbations, and theoretical modeling, we demonstrate that these asters likely generate an effective pushing force over short distances.

Valorisation of sea urchin (Paracentrotus lividus) gonads through canning

Fresh sea urchin (Paracentrotus lividus) gonads are a delicacy with short seasonal availability, very often heterogeneous in size and intrinsic characteristics. This study aimed to valorise this resource through the preparation of canned products (with/without Porphyra spp.) and evaluate their physicochemical and sensory quality (3–12 months). Canning contributed to a decrease in protein, K and most carotenoids contents; and a concentration of lipids, ash, Na and Se levels. A simulated 12-month ageing led to decrease the protein and β-carotene contents; and the Na and lutein levels concentration. The macroalgae addition resulted in an orange, darker and less soft product, with higher carbohydrates, Na, Se and carotenoids contents. A 25 g-dose contributes to significant daily intakes of protein (8–9%), EPA+DHA (47–53%), I (35–62%) and Se (30–47%). The products were commercially stable/sterile and had good sensory acceptance. Overall, canning constitutes a strategy to provide a nutritionally balanced product available all year-round.

Toxicity of organic solvents and surfactants to the sea urchin embryos

A comparative toxicity of widely applied organic solvents (methanol, ethanol, n-propanol, i-propanol, n-butanol, 2-butanol, i-butanol, t-butanol, 3-methoxy-3-methylbutanol-1 (MMB), ethylene glycol, diethylene glycol, 2-methoxyethanol, 2-ethoxyethanol, glycerol, ethyl acetate, acetonitrile, benzene, dioxane, dimethylformamide, dimethylacetamide, dimethylsulfoxide, 2-pyrrolidone, and N-methyl-2-pyrrolidone) and surfactants (PEG 300, PEG 6000, Tween 20, Tween 80, miramistin, and Cremophor EL) was studied using a sea urchin embryo model. Sea urchin embryo morphological alterations caused by the tested chemicals were described. The tested molecules affected P. lividus embryo development in a concentration-dependent manner. The observed phenotypic anomalies ranged from developmental delay and retardation of plutei growth to formation of aberrant blastules and gastrules, cleavage alteration/arrest, and embryo mortality. Discernible morphological defects were found after embryo exposure with common pharmaceutical ingredients, such as glycerol, Tween 80, and Cremophor EL. In general, solvents were less toxic than surfactants. PEG 6000 PEG 300, DMSO, ethanol, and methanol were identified as the most tolerable compounds with minimum effective concentration (MEC) values of 3.0–7.92 mg/mL. Previously reported MEC value of Pluronic F127 (4.0 mg/mL) fell within the same concentration range. Toxic effects of methanol, ethanol, DMSO, 2-methoxyethanol, 2-ethoxyethanol, Tween 20, and Tween 80 on P. lividus embryos correlated well with their toxicity obtained using other cell and animal models. The sea urchin embryos could be considered as an appropriate test system for toxicity assessment of solvents and surfactants for their further application as solubilizers of hydrophobic molecules in conventional in vitro cell-based assays and in vivo mammalian models. Nevertheless, to avoid adverse effect of a solubilizing agent in ecotoxicological and biological experiments, the preliminary assessment of its toxicity on a chosen test model would be beneficial.

Composite material in the sea urchin Cidaris rugosa: ordered and disordered micrometre-scale bicontinuous geometries.

The sponge-like biomineralized calcite materials found in echinoderm skeletons are of interest in terms of both structure formation and biological function. Despite their crystalline atomic structure, they exhibit curved interfaces that have been related to known triply periodic minimal surfaces. Here, we investigate the endoskeleton of the sea urchin Cidaris rugosa that has long been known to form a microstructure related to the Primitive surface. Using X-ray tomography, we find that the endoskeleton is organized as a composite material consisting of domains of bicontinuous microstructures with different structural properties. We describe, for the first time, the co-occurrenceof orderedsingle Primitive and singleDiamond structures and of a disordered structure within a single skeletal plate. We show that these structures can be distinguished by structural properties including solid volume fraction, trabeculae width and, to a lesser extent, interface area and mean curvature. Indoing so, we present a robust method that extracts interface areas and curvature integrals from voxelized datasets using the Steiner polynomial for parallel body volumes. We discuss these very large-scale bicontinuous structures in the context of their function, formation and evolution.

The positioning mechanics of microtubule asters in Drosophila embryo explants

Microtubule asters are essential in localizing the action of microtubules in processes including mitosis and organelle positioning. In large cells, such as the one-cell sea urchin embryo, aster dynamics are dominated by hydrodynamic pulling forces. However, in systems with more densely positioned nuclei such as the early Drosophila embryo, which packs around 6000 nuclei within the syncytium in a crystalline-like order, it is unclear what processes dominate aster dynamics. Here, we take advantage of a cell cycle regulation Drosophila mutant to generate embryos with multiple asters, independent from nuclei. We use an ex vivo assay to further simplify this biological system to explore the forces generated by and between asters. Through live imaging, drug and optical perturbations, and theoretical modeling, we demonstrate that these asters likely generate an effective pushing force over short distances.

Optimizing the biomass balance of macroalgae and sea urchins in kelp beds by removing the urchins

ABSTRACT In this study, we conducted sea urchin removal (SUR) experiments at two sites along Hokkaido’s southwestern and northwestern coasts in the Sea of Japan and one site along the northeast coast in the Sea of Okhotsk, to elucidate the biomass balance between macroalgae and sea urchins at different urchin barren sites. The experiments resulted in the restoration of macroalgal biomass at all sites and an increase in macroalgal diversity, indicating that it was the impact of high grazing pressure reducing the macroalgae, regardless of the particular experimental site. Notably, the recovery of the macroalgal biomass was characterized by increased kelp biomass, including Saccharina japonica var. religiosa and S. japonica var. ochotensis. In addition, negative correlations were observed between macroalgal biomass and sea urchin density. The lowest sea urchin density inducing an increase of macroalgal biomass was approximately the same in the reproductive growth phase (0.1 kg m−2) and in the vegetative growth phase (0.15 kg m−2) of Saccharina spp., the dominant kelps, regardless of the experimental site. These values can be helpful indicators for managing sea urchin density in order to restore macroalgae. In contrast, the maximum sea urchin density inducing the lowest macroalgal biomass varied markedly depending on the study site, indicating that different levels of kelp productivity are influenced by oceanographic conditions. Moreover, a strong positive correlation was observed between macroalgal biomass and sea urchins’ gonadosomatic index (GSI). Based on this correlation, macroalgal biomass supporting sea urchins with commercial value (GSI ≥ 15%) was estimated to be 1.6 kg m−2. This sea urchin density value is also a helpful indicator for managing macroalgal biomass and commercially valuable sea urchins. Moreover, this value will depend on the influence of the oceanographic environment of the site, particularly the nutrient conditions, on macroalgal productivity. In addition, we have also discussed how macroalgae and sea urchins can best coexist at the experimental sites.

Mesocosm study of PAC-modified clay effects on Karenia brevis cells and toxins, chemical dynamics, and benthic invertebrate physiology

Modified clay compounds are used globally as a method of controlling harmful algal blooms, and their use is currently under consideration to control Karenia brevis blooms in Florida, USA. In 1400 L mesocosm tanks, chemical dynamics and lethal and sublethal impacts of MC II, a polyaluminum chloride (PAC)-modified kaolinite clay, were evaluated over 72 h on a benthic community representative of Sarasota Bay, which included blue crab (Callinectes sapidus), sea urchin (Lytechinus variegatus), and hard clam (Mercenaria campechiensis). In this experiment, MC II was dosed at 0.2 g L-1 to treat bloom-level densities of K. brevis at 1 × 106 cells L-1. Cell removal in MC II-treated tanks was 57% after 8 h and 95% after 48 h. In the water column, brevetoxin analogs BTx-1 and BTx-2 were found to be significantly higher in untreated tanks at 24 and 48 h, while in MC II-treated tanks, BTx-3 was found to be higher at 48 h and BTx-B5 was found to be higher at 24 and 48 h. In MC II floc, we found no significant differences in BTx-1 or BTx-2 between treatments for any time point, while BTx-3 was found to be significantly higher in the MC II-treated tanks at 48 and 72 h, and BTx-B5 was higher in MC II-treated tanks at 24 and 72 h. Among various chemical dynamics observed, it was notable that dissolved phosphorus was consistently significantly lower in MC II tanks after 2 h, and that turbidity in MC II tanks returned to control levels 48 h after treatment. Dissolved inorganic carbon and total seawater alkalinity were significantly reduced in MC II tanks, and partial pressure of CO2 (pCO2) was significantly higher in the MC II-only treatment after 2 h. In MC II floc, particulate phosphorus was found to be significantly higher in MC II tanks after 24 h. In animals, lethal and sublethal responses to MC II-treated K. brevis did not differ from untreated K. brevis for either of our three species at any time point, suggesting MC II treatment at this dosage has negligible impacts to these species within 72 h of exposure. These results appear promising in terms of the environmental safety of MC II as a potential bloom control option, and we recommend scaling up MC II experiments to field trials in order to gain deeper understanding of MC II performance and dynamics in natural waters.

Myriad Mapping of nanoscale minerals reveals calcium carbonate hemihydrate in forming nacre and coral biominerals

Calcium carbonate (CaCO3) is abundant on Earth, is a major component of marine biominerals and thus of sedimentary and metamorphic rocks and it plays a major role in the global carbon cycle by storing atmospheric CO2 into solid biominerals. Six crystalline polymorphs of CaCO3 are known—3 anhydrous: calcite, aragonite, vaterite, and 3 hydrated: ikaite (CaCO3·6H2O), monohydrocalcite (CaCO3·1H2O, MHC), and calcium carbonate hemihydrate (CaCO3·½H2O, CCHH). CCHH was recently discovered and characterized, but exclusively as a synthetic material, not as a naturally occurring mineral. Here, analyzing 200 million spectra with Myriad Mapping (MM) of nanoscale mineral phases, we find CCHH and MHC, along with amorphous precursors, on freshly deposited coral skeleton and nacre surfaces, but not on sea urchin spines. Thus, biomineralization pathways are more complex and diverse than previously understood, opening new questions on isotopes and climate. Crystalline precursors are more accessible than amorphous ones to other spectroscopies and diffraction, in natural and bio-inspired materials.

Effects of thermal treatment on sea-urchin-like platinum nanoparticlese

The size and shape of metallic nanoparticles significantly influence the focus of light illumination and affect the plasmonic energy transformation process in plasmonic photocatalytic reactions. This study examined the effects of thermal treatment on the surface shape and plasmonic enhancement of Pt nanoparticles. SEM and XRD results indicate that the nanosized protruding thrusts on the surfaces of Pt sea urchin particles have a lower chemical potential and higher activity. They can be melted and reshaped at 600℃, which is much lower than their melting point of over 1700 ℃. The XPS results suggest that thermal treatment at 600℃ does not cause additional oxidation of the Pt nanocrystals. The samples not subjected to high-temperature treatment exhibited notable photocatalytic degradation of methylene blue in water under UV illumination. These results demonstrate that the surface features of Pt sea urchin nanoparticles play an essential role in the photocatalytic degradation of methylene blue under UV light. Larger particles treated at high temperatures exhibited reduced photocatalytic processing efficiency, whereas smaller particles treated at a lower temperature exhibited increased efficiency, indicating that the protruding sharp cones enhance plasmonic effects in photocatalytic reactions.

Mandible mechanical properties and composition of the larval Glossosoma boltoni (Trichoptera, Insecta)

Insect feeding structures, such as mandibles, interact with the ingesta (food or/and substrate) and can be adapted in morphology, composition of material and mechanical properties. The foraging on abrasive ingesta, as on algae covering rocks, is particularly challenging because the mandibles will be prone to wear and structural failure, thus suggesting the presence of mandibular adaptations to accompany this feeding behavior. Adaptations to this are well studied in the mouthparts of molluscs and sea urchins, but for insects there are large gaps in our knowledge. In this study, we investigated the mandibles of a grazing insect, the larvae of the trichopteran Glossosoma boltoni. Using scanning electron microscopy, wear was documented on the mandibles. The highest degree was identified on the medial surface of the sharp mandible tip. Using nanoindentation, the mechanical properties, such as hardness and Young’s modulus, of the medial and lateral mandible cuticles were tested. We found, that the medial cuticle of the tip was significantly softer and more flexible than the lateral one. These findings indicate that a self-sharpening mechanism is present in the mandibles of this species, since the softer medial cuticle is probably abraded faster than the harder lateral one, leading to sharp mandible tips. To investigate the origins of these properties, we visualized the degree of tanning by confocal laser scanning microscopy. The autofluorescence signal related to the mechanical property gradients. The presence of transition and alkaline earth metals by energy dispersive X-ray spectroscopy was also tested. We found Ca, Cl, Cu, Fe, K, Mg, Mn, P, S, Si, and Zn in the cuticle, but the content was very low and did not correlate with the mechanical property values.

Structural Basis for Multivalent MUC16 Recognition and Robust Anti-Pancreatic Cancer Activity of Humanized Antibody AR9.6.

Mucin-16 (MUC16) is a target for antibody-mediated immunotherapy in pancreatic ductal adenocarcinoma (PDAC) amongst other malignancies. The MUC16 specific monoclonal antibody AR9.6 has shown promise for PDAC immunotherapy and imaging. Here, we report the structural and biological characterization of the humanized AR9.6 antibody (huAR9.6). The structure of huAR9.6 was determined in complex with a MUC16 SEA (Sea urchin sperm, Enterokinase, Agrin) domain. Binding of huAR9.6 to recombinant, shed, and cell-surface MUC16 was characterized, and anti-PDAC activity was evaluated in vitro and in vivo. huAR9.6 bound a discontinuous, SEA domain epitope with an affinity of ~90 nM. Binding affinity depended on the specific SEA domain(s) present, and glycosylation enhanced affinity by 3-7-fold driven by favorable entropy and enthalpy and via distinct transition state thermodynamic pathways. Treatment with huAR9.6 reduced the in vitro growth, migration, invasion, and clonogenicity of MUC16-positive PDAC cells and patient-derived organoids (PDOs). HuAR9.6 blocked MUC16-mediated ErbB and AKT activation in PDAC cells, PDOs and patient-derived xenografts and induced antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity. More importantly, huAR9.6 treatment caused substantial PDAC regression in subcutaneous and orthotopic tumor models. The mechanism of action of huAR9.6 may depend on dense avid binding to homologous SEA domains on MUC16. The results of this study validate the translational therapeutic potential of huAR9.6 against MUC16-positive PDACs.

Investigating the diets and condition of Centrostephanus rodgersii (long-spined urchin) in barrens and macroalgae habitats in south-eastern Australia

Sea urchins are important herbivores that can graze macroalgae, creating ‘barren’ areas. These barrens are believed to offer little food for urchins due to the absence of attached macroalgae, leading to malnourishment as indicated by a low urchin gonad index. To investigate the diet and resultant gonad index of the long-spined sea urchin Centrostephanus rodgersii in New South Wales, Australia, we dissected 100 C. rodgersii individuals collected in macroalgae and barrens habitats along >470 km of SE NSW coastline. Diverse items were present in the digestive tracts of urchins from all habitats. These items included brown (42% barrens, 46% macroalgae), green (28% barrens, 42% macroalgae) and red algae (15% barrens, 12% macroalgae) and corallines (29% barrens, 37% macroalgae), molluscs (28% barrens, 29% macroalgae) and crustaceans (26% barrens, 22% macroalgae). There was no difference in urchin gut fullness between habitats (85% barrens, 90% macroalgae). Importantly, the gonad index only differed in macroalgae compared to barrens habitats at one location, with no differences detected at the other 4 locations. These results suggest that C. rodgersii has a diverse diet that is similar in both habitats, which could explain the similarities in gut fullness and gonad index. Our results suggest that C. rodgersii eat a broad diet including invertebrates and drift algae, and hence may not be malnourished in barrens. The finding of comparable gonad index between barrens and macroalgal areas further supports this conclusion. These findings challenge the prevailing perspective, indicating that sea urchins have sufficient food to survive and reproduce in different habitats.