Melt viscosity is central to choices of lunar processing technologies, including additive manufacturing, molten regolith electrolysis, and sintering processes, all of which may be central to future lunar exploration and in-situ resource utilization (ISRU). Here, we investigate the viscosity of a diverse set of lunar regolith simulants, specifically LX-M100, LX-T100, LMS-1, LHS-1, JSC-1A, EAC-1A, OPRL2N, OPRL2NT, OPRH2N, and OPRH3N. High-temperature concentric cylinder viscometry together with differential scanning calorimetry (DSC) / ‘Shift Factor’-derived viscosities were combined to describe viscosity across a broad temperature range, from superliquidus conditions to temperatures just above the glass transition (Tg). Selected fitted VFT-curves were compared with data measured by fast differential scanning calorimetry (FDSC). Post-viscometry glasses, obtained from reheated and then quenched samples, were analyzed using electron probe microanalysis (EPMA) and rock magnetic measurements. The results of these indicate post-viscometry heterogeneities (nano/microlites / liquid immiscibility) for some of the samples leading to their exclusion from calorimetric study but not impacting the interpretation of high-temperature viscosity measurements. Empirical and machine-learning-based viscosity models were assessed against experimental data, with the configurational-entropy model of Sehlke and Whittington (2016) performing robustly across the full range, while the recent gpvisc framework (Le Losq et al., 2025) yielded the lowest errors at high temperatures and near Tg These findings emphasize the importance of melt homogeneity and careful characterization to ensure accurate Tg-based viscosity predictions, vital for diverse lunar ISRU technologies especially under well-characterized redox conditions.

Chaetognaths, a phylum of enigmatic marine predators, present a significant challenge to phylogenetic reconstruction due to their uncertain evolutionary placement. While transcriptome analyses have suggested affinities with the Gnathifera clade, genomic data for this group remain scarce, hindering a comprehensive understanding of their evolution. Here, we present the first chromosome-level genome assembly of Flaccisagitta enflata, a species within the Aphragmophora order. The genome assembly includes 9 chromosome candidates with a total size of 794 Mb and a BUSCO score of 91.3% against the Metazoa lineage. This high-quality genome assembly provides a crucial resource for comparative genomic analyses within Chaetognatha and the broader Gnathifera clade, and it will facilitate investigations into chaetognath evolution and their phylogenetic relationships, addressing long-standing questions regarding their placement within the animal kingdom. ### Competing Interest Statement NG received a travel grant of 500$ from Oxford Nanopore to present their work at the conference of the European Society for Evolutionary Biology in Prague, 2022.

The phylum Placozoa remains one of the least explored among early-branching metazoan lineages. For over 130 years, this phylum had been represented by the single species Trichoplax adhaerens-an animal with the simplest known body plan (three cell layers without any organs) but complex behaviors. Recently, extensive sampling of placozoans across the globe and their subsequent genetic analysis have revealed incredible biodiversity with numerous cryptic species worldwide. However, only a few culture protocols are available to date, and all are for one species only. Here, we describe the breeding of four different species representing two placozoan genera: Trichoplax adhaerens, Trichoplax sp. H2, Hoilungia sp. H4, and Hoilungia hongkongensis originating from diverse biotopes. Our protocols allow to culture all species under comparable conditions. Next, we outlined various food sources and optimized strain-specific parameters enabling long-term culturing. These protocols can facilitate comparative analyses of placozoan biology and behaviors, which together will contribute to deciphering general principles of animal organization.

AbstractThe marine animal phylum Placozoa is characterized by a poorly explored cryptic biodiversity combined with very limited knowledge of their ecology. While placozoans are typically found as part of the epibenthos of coastal waters, known placozoan predators, namely small, shell‐less sea slugs belonging to the family Rhodopidae (Mollusca: Gastropoda: Heterobranchia), inhabit the interstitium of seafloor sediment. In order to gain further insights into this predator–prey relationship and to expand our understanding of placozoan ecological niches, we screened publicly available whole‐body metagenomic data from two rhodopid specimens collected from coastal sediments. Our analysis not only revealed the signatures of three previously unknown placozoan lineages in these sea slug samples but also enabled the assembly of three complete and two partial mitochondrial chromosomes belonging to four previously described placozoan genera, substantially extending the picture of placozoan biodiversity. Our findings further refine the molecular phylogeny of the Placozoa, corroborate the recently established taxonomic ranks in this phylum, and provide molecular support that known placozoan clades should be referred to as genera. We finally discuss the main finding of our study – the presence of placozoans in the sea floor sediment interstitium – in the context of their ecological, biological, and natural history implications.

Explosivity in erupting volcanoes is controlled by the degassing dynamics and the viscosity of the ascending magma in the conduit. Magma crystallisation enhances both heterogeneous bubble nucleation and increases in magma bulk viscosity. Nanolite crystallisation has been suggested to enhance such processes too, but in a noticeably higher extent. Yet the precise causes of the resultant strong viscosity increase remain unclear. Here we report experimental results for rapid nanolite crystallisation in natural silicic magma and the extent of the subsequent viscosity increase. Nanolite-free and nanolite-bearing rhyolite magmas were subjected to heat treatments, where magmas crystallised or re-crystallised oxide nanolites depending on their initial state, showing an increase of one order of magnitude as oxide nanolites formed. We thus demonstrate that oxide nanolites crystallisation increases magma bulk viscosity mainly by increasing the viscosity of its melt phase due to the chemical extraction of iron, whereas the physical effect of particle suspension is minor, almost negligible. Importantly, we further observe that this increase is sufficient for driving magma fragmentation depending on magma degassing and ascent dynamics.

Well-annotated and contiguous genomes are an indispensable resource for understanding the evolution, development, and metabolic capacities of organisms. Sponges, an ecologically important non-bilaterian group of primarily filter-feeding sessile aquatic organisms, are underrepresented with respect to available genomic resources. Here we provide a high-quality and well-annotated genome of Aphrocallistes vastus, a glass sponge (Porifera: Hexactinellida) that forms large reef structures off the coast of British Columbia (Canada). We show that its genome is approximately 80 Mb, small compared to most other metazoans, and contains nearly 2500 nested genes, more than other genomes. Hexactinellida is characterized by a unique skeletal architecture made of amorphous silicon dioxide (SiO2), and we identified 419 differentially expressed genes between the osculum, i.e. the vertical growth zone of the sponge, and the main body. Among the upregulated ones, mineralization-related genes such as glassin, as well as collagens and actins, dominate the expression profile during growth. Silicateins, suggested being involved in silica mineralization, especially in demosponges, were not found at all in the A. vastus genome and suggests that the underlying mechanisms of SiO2 deposition in the Silicea sensu stricto (Hexactinellida + Demospongiae) may not be homologous.

Mounting evidence suggests that animals and their associated bacteria interact via intricate molecular mechanisms, and it is hypothesized that disturbances to the microbiome influence animal development. Here, we show that the loss of a key photosymbiont (i.e., bleaching) upon shading correlates with a stark body-plan reorganization in the common aquarium cyanosponge Lendenfeldia chondrodes. The morphological changes observed in shaded sponges include the development of a thread-like morphology that contrasts with the flattened, foliose morphology of control specimens. The microanatomy of shaded sponges markedly differed from that of control sponges, with shaded specimens lacking a well-developed cortex and choanosome. Also, the palisade of polyvacuolar gland-like cells typical in control specimens was absent in shaded sponges. The morphological changes observed in shaded specimens are coupled with broad transcriptomic changes and include the modulation of signaling pathways involved in animal morphogenesis and immune response, such as the Wnt, transforming growth factor β (TGF-β), and TLR-ILR pathways. This study provides a genetic, physiological, and morphological assessment of the effect of microbiome changes on sponge postembryonic development and homeostasis. The correlated response of the sponge host to the collapse of the population of symbiotic cyanobacteria provides evidence for a coupling between the sponge transcriptomic state and the state of its microbiome. This coupling suggests that the ability of animals to interact with their microbiomes and respond to microbiome perturbations has deep evolutionary origins in this group.

The viscosity of molten Earth mantle has been determined for various redox states using very high temperature viscometry at 1 bar. The viscosity-temperature relationship of the most oxidised pyrolite studied here is comparable to that of a previous determination of the viscosity of a peridotite melt. For the first time, the effect of iron redox state on molten mantle viscosity has been determined by calorimetric analysis of the glass transition on extremely carefully characterised glassy samples quenched from conditions of variable fO2 using gas-mixing levitation. There is a clear trend of decreasing viscosity with increasing redox state over the range of Fe3+/sigma Fe. = 0.07-0.29. We also observe an increase in glass transition temperatures with increasing melt depolymerisation (i. e., higher NBO/T ratios). Both the negative oxidation dependence of viscosity and glass transition behaviour upon depolymerisation are contradictory to the trends observed from redox viscometry of more silica-rich melts but may be consistent with the broadly diminishing positive redox viscometry trends of melts with increasing basicity.

The pervasive association of demosponges with diverse bacterial communities has been a research topic since the late 1970s. Studies characterizing these bacterial communities have detected over 40 bacterial phyla associated with sponges and determined that sponge microbiomes differ from the bacterioplankton of the surrounding water and include sponges-specific bacterial associates. Despite the large body of information available on the diversity of the sponge microbiome, the molecular mechanisms used by sponges to interact with their bacterial symbionts and vice versa remain poorly understood. Yet, studying how sponges interact with their microbiomes is pivotal to understanding how these systems react to environmental changes and when these changes lead to a disruption of sponge-microbe associations. An important factor hampering the characterization of the sponge-microbiome molecular cross-talk mechanisms is the lack of broadly applicable molecular methods to, for instance, efficiently sequence meta-transcriptomes across a broad range of sponge species. To overcome this limitation, we implemented a hybrid-capture strategy capable of depleting both sponge and bacterial rRNA from total RNA extracts of highly divergent demosponges with microbiomes of different complexity. Our pan-demosponge rRNA depletion strategy allows for the efficient characterization of the metatranscriptome of diverse demosponge holobionts and the simultaneous quantification of gene expression in both the host and its microbiome.### Competing Interest StatementThe pan-demosponge bait set was designed by siTOOLs in collaboration with the authors. The authors did not receive any payments or will profit from future siTOOLs sales. siTOOLs was not involved in the analysis of the data generated or in writing the manuscript, but provided information and feedback used for the methods section.

The use of RNA sequencing (RNA-Seq) data and the generation of de novo transcriptome assemblies have been pivotal for studies in ecology and evolution. This is especially true for nonmodel organisms, where no genome information is available. In such organisms, studies of differential gene expression, DNA enrichment bait design and phylogenetics can all be accomplished with de novo transcriptome assemblies. Multiple tools are available for transcriptome assembly, but no single tool can provide the best assembly for all data sets. Therefore, a multi-assembler approach, followed by a reduction step, is often sought to generate an improved representation of the assembly. To reduce errors in these complex analyses while at the same time attaining reproducibility and scalability, automated workflows have been essential in the analysis of RNA-Seq data. However, most of these tools are designed for species where genome data are used as reference for the assembly process, limiting their use in nonmodel organisms. We present TransPi, a comprehensive pipeline for de novo transcriptome assembly, with minimum user input but without losing the ability of a thorough analysis. A combination of different model organisms, k-mer sets, read lengths and read quantities was used for assessing the tool. Furthermore, a total of 49 nonmodel organisms, spanning different phyla, were also analysed. Compared to approaches using single assemblers only, TransPi produces higher BUSCO completeness percentages, and a concurrent significant reduction in duplication rates. TransPi is easy to configure and can be deployed seamlessly using Conda, Docker and Singularity.

Placozoans are a promising model system to study fundamental regeneration processes in a morphologically and genetically very simple animal. We here provide a brief introduction to the enigmatic Placozoa and summarize the state of the art of animal handling and experimental manipulation possibilities.

Brasilicardin A ( 1 ) consists of an unusual anti / syn / anti -perhydrophenanthrene skeleton with a carbohydrate side chain and an amino acid moiety. It exhibits potent immunosuppressive activity, yet its mode of action differs from standard drugs that are currently in use. Further pre-clinical evaluation of this promising, biologically active natural product is hampered by restricted access to the ready material, as its synthesis requires both a low-yielding fermentation process using a pathogenic organism and an elaborate, multi-step total synthesis. Our semi-synthetic approach included a) the heterologous expression of the brasilicardin A gene cluster in different non-pathogenic bacterial strains producing brasilicardin A aglycone ( 5 ) in excellent yield and b) the chemical transformation of the aglycone 5 into the trifluoroacetic acid salt of brasilicardin A ( 1 a ) via a short and straightforward five-steps synthetic route. Additionally, we report the first preclinical data for brasilicardin A.

Methanol dried over powdered 4 Å molecular sieves can be used for a selective mono-de-O-acetylation of the phenolic acetyl group of the per-O-acetyl protected brasilicardin A carbohydrate side chain. This reaction opens a practical procedure for a synthetic access to derivates of the immunosuppressive and cytotoxic natural product brasilicardin A.

– Comparative measurements and analysis of 3D ERT and GPR results.– Evaluation of different ERT data processing methods.

From a morphological point of view, placozoans are among the most simple free-living animals. This enigmatic phylum is critical for our understanding of the evolution of animals and their cell types. Their millimeter-sized, disc-like bodies consist of only three cell layers that are shaped by roughly seven major cell types. Placozoans lack muscle cells and neurons but are able to move using their ciliated lower surface and take up food in a highly coordinated manner. Intriguingly, the genome of Trichoplax adhaerens, the founding member of the enigmatic phylum, has disclosed a surprising level of genetic complexity. Moreover, recent molecular and functional investigations have uncovered a much larger, so-far hidden cell-type diversity. Here, we have extended the microanatomical characterization of a recently described placozoan species—Hoilungia hongkongensis. In H. hongkongensis, we recognized the established canonical three-layered placozoan body plan but also came across several morphologically distinct and potentially novel cell types, among them novel gland cells and “shiny spheres”-bearing cells at the upper epithelium. Thus, the diversity of cell types in placozoans is indeed higher than anticipated.

Foraminifera are single-celled eukaryotes (protists) of large ecological importance, as well as environmental and paleoenvironmental indicators and biostratigraphic tools. In addition, they are capable of surviving in anoxic marine environments where they represent a major component of the benthic community. However, the cellular adaptations of Foraminifera to the anoxic environment remain poorly constrained. We sampled an oxic-anoxic transition zone in marine sediments from the Namibian shelf, where the generaBolivinaandStainforthiadominated the Foraminifera community, and use metatranscriptomics to characterize Foraminifera metabolism across the different geochemical conditions. Relative Foraminifera gene expression in anoxic sediment increased an order of magnitude, which was confirmed in a 10-day incubation experiment where the development of anoxia coincided with a 20-40-fold increase in the relative abundance of Foraminifera protein encoding transcripts, attributed primarily to those involved in protein synthesis, intracellular protein trafficking, and modification of the cytoskeleton. This indicated that many Foraminifera were not only surviving but thriving, under the anoxic conditions. The anaerobic energy metabolism of these active Foraminifera was characterized by fermentation of sugars and amino acids, fumarate reduction, and potentially dissimilatory nitrate reduction. Moreover, the gene expression data indicate that under anoxia Foraminifera use the phosphogen creatine phosphate as an ATP store, allowing reserves of high-energy phosphate pool to be maintained for sudden demands of increased energy during anaerobic metabolism. This was co-expressed alongside genes involved in phagocytosis and clathrin-mediated endocytosis (CME). Foraminifera may use CME to utilize dissolved organic matter as a carbon and energy source, in addition to ingestion of prey cells via phagocytosis. These anaerobic metabolic mechanisms help to explain the ecological success of Foraminifera documented in the fossil record since the Cambrian period more than 500 million years ago.

The membrane-associated progesterone receptor (MAPR) family consists of heme-binding proteins containing a cytochrome b(5) (cytb(5)) domain characterized by the presence of a MAPR-specific interhelical insert region (MIHIR) between helices 3 and 4 of the canonical cytb5-domain fold. Animals possess three MAPR genes (PGRMC-like, Neuferricin and Neudesin). Here we show that all three animal MAPR genes were already present in the common ancestor of the opisthokonts (comprising animals and fungi as well as related single-celled taxa). All three MAPR genes acquired extensions C-terminal to the cytb(5) domain, either before or with the evolution of animals. The archetypical MAPR protein, progesterone receptor membrane component 1 (PGRMC1), contains phosphorylated tyrosines Y139 and Y180. The combination of Y139/Y180 appeared in the common ancestor of cnidarians and bilaterians, along with an early embryological organizer and synapsed neurons, and is strongly conserved in all bilaterian animals. A predicted protein interaction motif in the PGRMC1 MIHIR is potentially regulated by Y139 phosphorylation. A multilayered model of animal MAPR function acquisition includes some pre-metazoan functions (e.g., heme binding and cytochrome P450 interactions) and some acquired animal specific functions that involve regulation of strongly conserved protein interaction motifs acquired by animals (Metazoa). This study provides a conceptual framework for future studies, against which especially PGRMC1 ' s multiple functions can perhaps be stratified and functionally dissected.

Brasilicardin A (BraA) is a promising immunosuppressive compound produced naturally by the pathogenic bacterium Nocardia terpenica IFM 0406. Heterologous host expression of brasilicardin gene cluster showed to be efficient to bypass the safety issues, low production levels and lack of genetic tools related with the use of native producer. Further improvement of production yields requires better understanding of gene expression regulation within the BraA biosynthetic gene cluster (Bra-BGC); however, the only so far known regulator of this gene cluster is Bra12. In this study, we discovered the protein LysRNt, a novel member of the LysR-type transcriptional regulator family, as a regulator of the Bra-BGC. Using in vitro approaches, we identified the gene promoters which are controlled by LysRNt within the Bra-BGC. Corresponding genes encode enzymes involved in BraA biosynthesis as well as the key Bra-BGC regulator Bra12. Importantly, we provide in vivo evidence that LysRNt negatively affects production of brasilicardin congeners in the heterologous host Amycolatopsis japonicum. Finally, we demonstrate that some of the pathway related metabolites, and their chemical analogs, can interact with LysRNt which in turn affects its DNA-binding activity.

Bacterial symbionts are integral to the health and homeostasis of invertebrate hosts. Notably, members of the Rickettsiales genus Wolbachia influence several aspects of the fitness and evolution of their terrestrial hosts, but few analogous partnerships have been found in marine systems. We report here the genome, phylogenetics, and biogeography of a ubiquitous and novel Rickettsiales species that primarily associates with marine organisms. We previously showed that this bacterium was found in scleractinian corals, responds to nutrient exposure, and is associated with reduced host growth and increased mortality. This bacterium, like other Rickettsiales, has a reduced genome indicative of a parasitic lifestyle. Phylogenetic analysis places this Rickettsiales within a new genus we define as "Candidatus Aquarickettsia." Using data from the Earth Microbiome Project and SRA databases, we also demonstrate that members of "Ca. Aquarickettsia" are found globally in dozens of invertebrate lineages. The coral-associated "Candidatus A. rohweri" is the first finished genome in this new clade. "Ca. A. rohweri" lacks genes to synthesize most sugars and amino acids but possesses several genes linked to pathogenicity including Tlc, an antiporter that exchanges host ATP for ADP, and a complete Type IV secretion system. Despite its inability to metabolize nitrogen, "Ca. A. rohweri" possesses the NtrY-NtrX two-component system involved in sensing and responding to extracellular nitrogen. Given these data, along with visualization of the parasite in host tissues, we hypothesize that "Ca. A. rohweri" reduces coral health by consuming host nutrients and energy, thus weakening and eventually killing host cells. Last, we hypothesize that nutrient enrichment, which is increasingly common on coral reefs, encourages unrestricted growth of "Ca. A. rohweri" in its host by providing abundant N-rich metabolites to be scavenged.

SUMMARY Understanding the temporal changes of the Earth’s magnetic field intensity is one of the main goals of modern palaeomagnetism. For most palaeointensity methods to yield reliable results, the magnetic minerals must obey a set of rules. One of these rules is the additivity of partial thermal (TRM) or anhysteretic remanent magnetizations (ARM). Additivity was previously shown for partial TRM in single-domain particles and more generally for ARMs. Additivity between these two low-field remanences, however, has not been investigated, yet. This paper presents a series of rock magnetic experiments on natural low Ti titanomagnetites (Curie temperature between 534 °C and 561 °C) examining the effects of high temperatures on alternating field (AF) demagnetization and acquisition of an ARM. One of our sample sets comes from a borehole drilled through the impact melt sheet of the Manicouagan crater (Canada), the other from the Rocche Rosse lava flow on the island of Lipari (Italy). Hysteresis parameters indicate the magnetic carriers in the pseudo-single-domain range showing no evidence for oxidation. Thermal demagnetization at 300 °C and 500 °C before AF demagnetization shifts the coercivity spectra towards higher fields. AF demagnetization experiments at 500 °C show a significant (by a factor between 1.4 and >7.6) reduction in median destructive field and a shift towards lower coercivities. A linear relationship was found between the peak magnetic field required to demagnetize a fraction of a full TRM of a sample at a specific temperature and the one necessary to demagnetize the same fraction at room temperature after heating to that temperature. The comparison of full ARM and partial TRM at successively higher temperatures with a hybrid hTARM reveals that combined additivity between the two kinds of remanences is fulfilled. These results open the possibility to demagnetize highly coercive minerals, such as hematite and goethite, which is often not achievable at elevated temperatures. Furthermore, the additivity of TRM and ARM remanences may be used to develop novel hybrid TRM/ARM palaeointensity methods for samples, where heating is problematic (e.g. in meteorites).