Symbiont transmission in marine sponges: reproduction, development, and metamorphosis.

Marine sponges (phylum Porifera) form symbioses with diverse microbial communities that can be transmitted between generations through their developmental stages. Here, we integrate embryology and microbiology to review how symbiotic microorganisms are transmitted in this early-diverging lineage. We describe that vertical transmission is widespread but not universal, that microbes are vertically transmitted during a select developmental window, and that properties of the developmental microbiome depends on whether a species is a high or low microbial abundance sponge. Reproduction, development, and symbiosis are thus deeply rooted, but why these partnerships form remains the central and elusive tenet of these developmental symbioses.

Improved landfill leachate quality using ozone, UV solar radiation, hydrogen peroxide, persulfate and adsorption processes.

Advanced oxidation processes based on ozonation, UV solar radiation, hydrogen peroxide, and persulfate were tested for the complete treatment of a specific landfill leachate. As a post-treatment of the advanced oxidation process, an additional adsorption process was carried out using a natural zeolite. Application of the UVsolar/O3/H2O2 process required 140 min of irradiation and the consumption of 0.67 g/L H2O2 to eliminate 56% and 17% of colour and chemical oxygen demand (COD), respectively. When adding persulfate to the system (UVsolar/O3/H2O2/S2O8-2), colour and COD were reduced by 29% and 77%, respectively, with a 0.2 g/L concentration of S2O8-2 requiring 250 min of irradiation time. In an experimental run with landfill leachate, adsorption post-treatment with a natural zeolite resulted in 36%, 99%, and 18% total reductions in COD, ammonium, and chloride, respectively. In another experimental run, adsorption with a zeolite was used as a pre-treatment for the advanced oxidation process (UVsolar/O3/H2O2 and UVsolar/O3/H2O2/S2O8-2). The results were similar to those for adsorption post-treatment, with 30%, 90%, and 20% total reductions in COD, ammonium, and chloride, respectively. Despite improvements, some measured parameters of treated landfill leachate still exceeded Chilean regulations for water quality. Furthermore, Lactuca sativa seeds did not germinate after irrigation with 100% treated landfill leachate or dilutions above 3%. Finally, EC50 values were enhanced during treatment, going from 0.002% for raw landfill leachate to 1.179% after the adsorption process.

Chemoselective Reactions for the Synthesis of Glycoconjugates from Unprotected Carbohydrates.

Glycobiology is the comprehensive biological investigation of carbohydrates. The study of the role and function of complex carbohydrates often requires the attachment of carbohydrates to surfaces, their tagging with fluorophores, or their conversion into natural or non-natural glycoconjugates, such as glycopeptides or glycolipids. Glycobiology and its "omics", glycomics, require easy and robust chemical methods for the construction of these glycoconjugates. This review gives an overview of the rapidly expanding field of chemical reactions that selectively convert unprotected carbohydrates into glycoconjugates through the anomeric position. The discussion is divided in terms of the anomeric bond type of the newly formed glycoconjugates, including O-, N-, S-, and C-glycosides.

Folding Topology of a Short Coiled-Coil Peptide Structure Templated by an Oligonucleotide Triplex.

The rational design of a well-defined protein-like tertiary structure formed by small peptide building blocks is still a formidable challenge. By using peptide-oligonucleotide conjugates (POC) as building blocks, we present the self-assembly of miniature coiled-coil α-helical peptides guided by oligonucleotide duplex and triplex formation. POC synthesis was achieved by copper-free alkyne-azide cycloaddition between three oligonucleotides and a 23-mer peptide, which by itself exhibited multiple oligomeric states in solution. The oligonucleotide domain was designed to furnish a stable parallel triplex under physiological pH, and to be capable of templating the three peptide sequences to constitute a small coiled-coil motif displaying remarkable α-helicity. The formed trimeric complex was characterized by ultraviolet thermal denaturation, gel electrophoresis, circular dichroism (CD) spectroscopy, small-angle X-ray scattering (SAXS), and molecular modeling. Stabilizing cooperativity was observed between the trimeric peptide and the oligonucleotide triplex domains, and the overall molecular size (ca. 12 nm) in solution was revealed to be independent of concentration. The topological folding of the peptide moiety differed strongly from those of the individual POC strands and the unconjugated peptide, exclusively adopting the designed triple helical structure.

A Proposal for the Evolution of Cathepsin and Silicatein in Sponges.

Cathepsins are enzymes capable of degrading proteins intracellularly. They occur ubiquitously in opisthokonts, but their potential to provide insight across the evolutionary transition from protists to metazoans remains poorly investigated. Here, we explore the evolution of cathepsins using comparative analyses of transcriptomic datasets, focusing on both, protists (closely related to metazoans), and early divergent animals (i.e., sponges). We retrieved DNA sequences of nine cathepsin types (B, C, D, F, H, L, O, Z, and silicatein) in the surveyed taxa. In choanoflagellates, only five types (B, C, L, O, Z) were identified, all of them being also found in sponges, indicating that while all cathepsins present in protists were conserved across metazoan lineages, cathepsins F and H (and probably D) are metazoan acquisitions. The phylogeny of cysteine protease cathepsins (excluding cathepsin D) revealed two major lineages: lineage B (cathepsins B and C) and lineage L (cathepsins F, H, L, O, Z). In the latter lineage, a mutation at the active site of cathepsin L gave rise to silicatein, an enzyme exclusively known to date from siliceous sponges and involved in the production of their silica spicules. However, we found that several sponges with siliceous spicules did not express silicatein genes and that, in contrast, several aspiculate sponges did contain silicatein genes. Our results suggest that the ability to silicify may have evolved independently within sponges, some of them losing this capacity secondarily. We also show that most phylogenies based on cathepsin and silicatein genes (except for that of cathepsin O) failed to recover the major lineages of sponges.

[Validation of the Rockall score in elderly patients with non variceal upper gastrointestinal bleeding in a third level general hospital]. / Validación del score de Rockall en pacientes adultos mayores con hemorragia digestiva alta no variceal en un hospital general de tercer nivel.

OBJECTIVE: To validate the Rockall score in elderly patients with non variceal upper gastrointestinal bleeding, in terms of mortality and recurrent bleeding at 30 days follow-up. MATERIAL AND METHODS: Patients older than 60 year-old, with non variceal upper gastrointestinal bleeding demonstrated by endoscopy, who were attended in a third level general hospital from June 2009 to June 2013, were included. Data was analyzed with the ROC (Receiver Operating Characteristic) curve. The area under curve was obtained to assess the Rockall score. RESULTS: One hundred ninety patients were included, 64.2% were males, with an average age of 74 years old. Overall mortality was 16.8% and respiratory failure with second bleeding episode was the most common cause of death (34.3% and 31.3% respectively). 5.52% of patients presented a second bleeding episode. A transfusion of more than 2 blood-packs was needed in the 24.7% of the patients. The area under ROC curve using the Rockall score was 0,76 (IC: 0.68-0.84) for mortality risk, 0.71 (IC: 0.55-0.88) for the risk of rebleeding and 0.66 (IC: 0.58-0.74) for needing a more than 2 blood-packs transfusion. CONCLUSIONS: Rockall score is a good predictor for mortality and rebleeding during the 30 day-period after a non-variceal bleeding episode in elderly patients. The best sensibility and specificity was obtained with the scores of 5 and 6, respectively.

ß-Cyclodextrin-bearing gold glyconanoparticles for the development of site specific drug delivery systems.

Three novel gold nanoparticles containing multiple long, flexible linkers decorated with lactose, ß-cyclodextrin, and both simultaneously have been prepared. The interaction of such nanoparticles with ß-d-galactose-recognizing lectins peanut agglutinin (PNA) and human galectin-3 (Gal-3) was demonstrated by UV-vis studies. Gal-3 is well-known to be overexpressed in several human tumors and can act as a biorecognizable target. This technique also allowed us to estimate their loading capability toward the anticancer drug methotrexate (MTX). Both results make these glyconanoparticles potential site-specific delivery systems for anticancer drugs.

New and rare sponges from the deep shelf of the Alboran Island (Alboran Sea, Western Mediterranean).

The sponge fauna from the deep shelf (70 to 200 m) of the Alboran Island (Alboran Sea, Western Mediterranean) was investigated using a combination of ROV surveys and collecting devices in the frame of the EC LIFE+ INDEMARES Grant aimed to designate marine areas of the Nature 2000 Network within Spanish territorial waters. From ROV surveys and 351 examined specimens, a total of 87 sponge species were identified, most belonging in the Class Demospongiae, and one belonging in the Class Hexactinellida. Twenty six (29%) species can be regarded as either taxonomically or faunistically relevant. Three of them were new to science (Axinella alborana nov. sp.; Axinella spatula nov. sp.; Endectyon filiformis nov. sp.) and 4 others were Atlantic species recorded for the first time in the Mediterranean Sea (Jaspis eudermis Lévi & Vacelet, 1958; Hemiasterella elongata Topsent, 1928; Axinella vellerea Topsent, 1904; Gelliodes fayalensis Topsent, 1892). Another outstanding finding was a complete specimen of Rhabdobaris implicata Pulitzer-Finali, 1983, a species only known from its holotype, which had entirely been dissolved for its description. Our second record of the species has allowed a neotype designation and a restitution of the recently abolished genus Rhabdobaris Pulitzer-Finally, 1983, also forcing a slight modification of the diagnosis of the family Bubaridae. Additionally, 12 species were recorded for the first time from the shelf of the Alboran Island, including a few individuals of the large hexactinellid Asconema setubalense Kent, 1877 that provided the second Mediterranean record of this "North Atlantic" hexactinellid. ROV explorations also revealed that sponges are an important component of the deep-shelf benthos, particularly on rocky bottoms, where they make peculiar sponge gardens characterized by a wide diversity of small, erect species forming a dense "undergrowth" among a scatter of large sponges and gorgonians. The great abundance and the taxonomic singularities of the sponge fauna occurring in these deep-shelf bottoms strongly suggest these habitats to be considered within the environmental protection of the Nature 2000 Network.

Silica-associated proteins from hexactinellid sponges support an alternative evolutionary scenario for biomineralization in Porifera.

Metazoans use silicon traces but rarely develop extensive silica skeletons, except for the early-diverging lineage of sponges. The mechanisms underlying metazoan silicification remain incompletely understood, despite significant biotechnological and evolutionary implications. Here, the characterization of two proteins identified from hexactinellid sponge silica, hexaxilin and perisilin, supports that the three classes of siliceous sponges (Hexactinellida, Demospongiae, and Homoscleromorpha) use independent protein machineries to build their skeletons, which become non-homologous structures. Hexaxilin forms the axial filament to intracellularly pattern the main symmetry of the skeletal parts, while perisilin appears to operate in their thickening, guiding extracellular deposition of peripheral silica, as does glassin, a previously characterized hexactinellid silicifying protein. Distant hexaxilin homologs occur in some bilaterians with siliceous parts, suggesting putative conserved silicifying activity along metazoan evolution. The findings also support that ancestral Porifera were non-skeletonized, acquiring silica skeletons only after diverging into major classes, what reconciles molecular-clock dating and the fossil record.

Microbiome changes through the ontogeny of the marine sponge Crambe crambe.

BACKGROUND: Poriferans (sponges) are highly adaptable organisms that can thrive in diverse marine and freshwater environments due, in part, to their close associations with internal microbial communities. This sponge microbiome can be acquired from the surrounding environment (horizontal acquisition) or obtained from the parents during the reproductive process through a variety of mechanisms (vertical transfer), typically resulting in the presence of symbiotic microbes throughout all stages of sponge development. How and to what extent the different components of the microbiome are transferred to the developmental stages remain poorly understood. Here, we investigated the microbiome composition of a common, low-microbial-abundance, Atlantic-Mediterranean sponge, Crambe crambe, throughout its ontogeny, including adult individuals, brooded larvae, lecithotrophic free-swimming larvae, newly settled juveniles still lacking osculum, and juveniles with a functional osculum for filter feeding. RESULTS: Using 16S rRNA gene analysis, we detected distinct microbiome compositions in each ontogenetic stage, with variations in composition, relative abundance, and diversity of microbial species. However, a particular dominant symbiont, Candidatus Beroebacter blanensis, previously described as the main symbiont of C. crambe, consistently occurred throughout all stages, an omnipresence that suggests vertical transmission from parents to offspring. This symbiont fluctuated in relative abundance across developmental stages, with pronounced prevalence in lecithotrophic stages. A major shift in microbial composition occurred as new settlers completed osculum formation and acquired filter-feeding capacity. Candidatus Beroebacter blanensis decreased significatively at this point. Microbial diversity peaked in filter-feeding stages, contrasting with the lower diversity of lecithotrophic stages. Furthermore, individual specific transmission patterns were detected, with greater microbial similarity between larvae and their respective parents compared to non-parental conspecifics. CONCLUSIONS: These findings suggest a putative vertical transmission of the dominant symbiont, which could provide some metabolic advantage to non-filtering developmental stages of C. crambe. The increase in microbiome diversity with the onset of filter-feeding stages likely reflects enhanced interaction with environmental microbes, facilitating horizontal transmission. Conversely, lower microbiome diversity in lecithotrophic stages, prior to filter feeding, suggests incomplete symbiont transfer or potential symbiont digestion. This research provides novel information on the dynamics of the microbiome through sponge ontogeny, on the strategies for symbiont acquisition at each ontogenetic stage, and on the potential importance of symbionts during larval development.

Highly efficient grafting of hetero-complementary amidinium and carboxylate hydrogen-bonding/ionic pairs onto polymer surfaces.

We describe a grafting methodology, based on thiol-fluoroarene chemistry, to efficiently incorporate complementary hydrogen-bonding carboxylate and amidinium groups into polymer backbones. The process was optimized both in solution and on the surface of processed films, with the aim to produce materials showing hetero-complementary adhesion.

Deep metazoan phylogeny: when different genes tell different stories.

Molecular phylogenetic analyses have produced a plethora of controversial hypotheses regarding the patterns of diversification of non-bilaterian animals. To unravel the causes for the patterns of extreme inconsistencies at the base of the metazoan tree of life, we constructed a novel supermatrix containing 122 genes, enriched with non-bilaterian taxa. Comparative analyses of this supermatrix and its two non-overlapping multi-gene partitions (including ribosomal and non-ribosomal genes) revealed conflicting phylogenetic signals. We show that the levels of saturation and long branch attraction artifacts in the two partitions correlate with gene sampling. The ribosomal gene partition exhibits significantly lower saturation levels than the non-ribosomal one. Additional systematic errors derive from significant variations in amino acid substitution patterns among the metazoan lineages that violate the stationarity assumption of evolutionary models frequently used to reconstruct phylogenies. By modifying gene sampling and the taxonomic composition of the outgroup, we were able to construct three different yet well-supported phylogenies. These results show that the accuracy of phylogenetic inference may be substantially improved by selecting genes that evolve slowly across the Metazoa and applying more realistic substitution models. Additional sequence-independent genomic markers are also necessary to assess the validity of the phylogenetic hypotheses.

Poly(amido amine)-based mannose-glycodendrimers as multielectron redox probes for improving lectin sensing.

An easy-to-prepare series of electroactive poly(amido amine) (PAMAM)-based dendrimers of generations G0 to G2 having mannopyranosylferrocenyl moieties in the periphery to detect carbohydrate-protein interactions is reported. The synthesis involved the functionalization of the PAMAM surface with azidomethylferrocenyl groups and subsequent coupling of mannoside units by the Cu(I)-catalyzed Huisgen reaction. The binding affinity of the series of electroactive glycodendrimers was studied by isothermal titration calorimetry (ITC) and differential pulse voltammetry (DPV). Upon complexation of the glycodendrimers conjugates with prototypical concanavalin A (Con A), voltammograms showed a decrease of the peak current. Such dendrimers showed a notable improvement of redox sensing abilities toward Con A when compared with mono- and divalent analogues, based on both the glycoside multivalent and ferrocene dendritic effects.

[Risk factors associated to mortality by upper GI bleeding in patients from a public hospital. A case control study]. / Factores de riesgo asociados a la mortalidad por hemorragia digestiva alta en pacientes de un hospital público. Estudio caso control.

OBJECTIVE: Identify and establish risk factors associated with mortality secondary to upper gastrointestinal bleeding up to 30 days after the episode, at the Hospital Nacional Cayetano Heredia. MATERIAL AND METHODS: A retrospective analytic observational case-control study was made with a case: control proportion of 1:3, analyzing 180 patient from which 135 were the controls and 45 the cases. It was determined by biological plausibility as potential risk factors to 14 variables, with which were performed bivariate and multivariate logistic analyses. RESULTS: It was found in the bivariate logistic analysis as variables statistically related to mortality: age (OR=1.02), hematemesis (OR=2.57), in-hospital upper gastrointestinal bleeding (OR=4), cirrhosis (OR=2.67), malignancy (OR=5,37), admittance to intensive care unit/Shock-Trauma (OR=9.29), Rockall score greater than 4 (OR=19.75), rebleeding (OR=5.65), and number of packed red blood cell transfusions(OR=1.22). While in the multivariate logistic analysis, the only variables statistically related to mortality were: malignancy (OR=5.35), admittance to intensive care unit/Shock-Trauma (OR=8.29), and Rockall score greater than 4 (OR=8.43). CONCLUSIONS: The factors that increase the risk of mortality in patient with upper gastrointestinal bleeding are: Rockall score greater than 4, admittance to intensive care unit/Shock-Trauma, rebleeding, in-hospital upper gastrointestinal bleeding, cirrhosis, hematemesis, malignancy, the number of red blood cell transfusion, and age.

[Frequency and mortality by rebleeding in cirrhotic patients treated for bleeding esophagic varices in two hospitals in Lima Peru during years 2009 to 2011]. / Frecuencia y mortalidad de resangrado en pacientes cirróticos tratados por hemorragia de várices esofágicas en dos hospitales de Lima-Perú durante los años 2009-2011.

UNLABELLED: During the first 6 weeks after a variceal hemorrhage there is a 30-40% of probability of recurrence and those who rebleed 20- 30% die. Passed this period, the risk of rebleeding is of 60% and reaches a mortality of 60-70% in two years without treatment. OBJECTIVE: Describe the frequency of rebleeding and mortality due to rebleeding in cirrhotic patients treated for variceal hemorrhage at Endoscopic Centers of Hospital Nacional Cayetano Heredia, Lima, Peru and Hospital Nacional Arzobispo Loayza, Lima, Peru during the years 2009-2011. MATERIALS AND METHODS: The study type is a transversal, periodic and retrospective one in which were included 176 cirrhotic patients older than 14 years who have bleed for esophageal varices and that have received endoscopic therapy. The instruments used were a data sheet with all the information obtained from the clinical chart of each patient, the CHILD score to assess severity of hepatic disease, endoscopic informs and phone calls. RESULTS: The frequency of rebleeding before 6 weeks was 32.20% (56 patients). Also, the frequency of rebleeding after that time was 22.56% (37 patients). There was a mortality rate of 5.70% (10 patients) and a mortality rate due to rebleeding of 13.33% (6 patients). CONCLUSIONS: Variceal hemorrhage is an important cause of mobimortality in peruvian people. The frequency of rebleeding and mortality due to rebleeding resulted slightly lower than in other countries.

[The usefulness of the Baylor score in patients with upper gastrointestinal bleeding in a hospital in Lima, Peru]. / Utilidad del score de Baylor en pacientes con hemorragia digestiva alta en un hospital de Lima, Perú

OBJECTIVE: To demonstrate the usefulness of the Baylor score in patients with upper gastrointestinal bleeding (UGB) due to peptic ulcer, in terms of mortality and recurrent bleeding at 30 days follow-up. MATERIAL AND METHODS: This study has collected information prospectively into a registration form from medical histories. Patients included were those who came to the "Hospital Nacional Cayetano Heredia", Lima, Peru, in the period between June 2009 and May 2011, with UGB due to peptic ulcer demonstrated by endoscopy. The data was analyzed with the ROC curve (Receiver Operating Characteristic). RESULTS: We included 181 from 380 patients with UGB (47.63%), 74% were male, mean age 56 years old; 20 patients died during follow-up: 8 due to gastrointestinal bleeding and 12 from other causes, 10 patients had recurrence of bleeding during a 30 days follow-up. When performing the analysis of the ROC curve with the Baylor score and mortality, it was reported IC 95% value of 0.89 (0.83-0.95), with recurrence of bleeding an IC 95% value of 0.81 (0.68-0.93) and need for transfusion of over two globular packages an IC 95% value of 0.79 (0.70-0.86). CONCLUSIONS: Baylor score is a good predictor of mortality and recurrence of bleeding during the first 30 days after a first episode of an upper gastrointestinal bleeding and an acceptable predictor of the need to transfuse more than two globular packages during hospitalization.

A giant foraminifer that converges to the feeding strategy of carnivorous sponges: Spiculosiphon oceana sp. nov. (Foraminifera, Astrorhizida).

The foraminifer Spiculosiphon oceana sp. nov. is a giant (>4 cm) agglutinated astrorhizid, which makes the second known species of this unusual genus and its first Mediterranean record. It has a peculiar stalked, capitate, monothalamous test. Bleach digestion and X-ray microanalysis indicated the test to be made exclusively of siliceous sponge spicules agglutinated in organic cement. The organism stands on a hollow, 4 cm long, 0.5 cm thick stalk built with highly selected, long and thin spicule fragments, tightly cemented together in parallel to the main axis of the stalk. The proximal end of the stalk is closed and slightly expanded into a bulb-like structure, designed to penetrate between the sand grains and maintaining the test upright while avoiding a permanent attachment to the substratum. The distal stalk end becomes a hollow, globe-like structure that contains the main protoplasm. The globelike region is built with loosely agglutinated and irregularly-shaped spicules, allowing extrusion of the pseudopodia through the cavities between the spicules. The globelike structure also serves as an anchoring basis, from which long and thin, solid tracts protrude radially to make a spherical crown that attains about 4 mm in total diameter. The radiating tracts are built with highly selected aciculate spicule fragments held together with a translucent organic cement. They provide skeletal support for the extension of a crown of pseudopodia into the water column. This arrangement is thought to enhance the chances of the pseudopodia to contact demersal planktonic prey. In summary, Spiculosiphon species collect and arrange sponge spicules with high selectivity to recreate a body morphology that strongly converges to that of some carnivorous sponges, which allows these predatory foraminifera to exploit a prey capturing strategy similar to that of the carnivorous sponges. This idea is also consistent with our report of an additional, yet undetermined, Spiculosiphon species occurring in the same sublittoral Mediterranean cave where carnivorous sponges were first discovered.

Use of vinasse and coffee waste as chelating agent of photo-Fenton landfill leachate treatment.

This research studies the use of vinasse (VS) coming from Pisco and caffeic acid (Caa) from solid coffee waste as chelating agents of this process, to carry out a photo-Fenton process using UVc lamps of 254-nm wavelength for 60 min, at the natural pH of the landfill leachate (8.9). Without the chelating agent, there was a removal of UV 254 and COD of 54.2% and 54.7%, respectively, when the photo-Fenton reaction was carried out at pH 3; at pH 6, the removal of UV 254 and COD was 13.1% and 39.2%, respectively, and at pH 8.9, the elimination of UV 254 and COD was 10.8% and 16.1%, respectively. When Caa was used in the landfill leachate (LL) for the photo-catalytic processes carried out at pH 8.9, a removal of 24.1%, 43.0%, and 47.4% of UV 254 was obtained using 5 mg/L, 50 mg/L, and 100 mg/L of Caa. The removal of UV 254 was 27.3%, 30.7%, and 36.3% using 5 mg/L, 50 mg/L, and 100 mg/L of VS, respectively, and the removal of COD was 32.2%, 35.4%, and 39.2% using 5 mg/L, 50 mg/L, and 100 mg/L of VS, respectively. When Caa was used in the LL at pH 8.9, the concentration of total Fe went from 37.5 to 33.2, from 40.2 to 36.8, and from 45.2 to 42.1, using 5 mg/L, 50 mg/L, and 100 mg/L of caffeic acid, respectively. Using VS in the LL at pH 8.9, the concentration of total Fe along the run went from 35.1 to 32.2, from 39.4 to 34.8, and from 42.1 to 40.2, using 5 mg/L, 50 mg/L, and 100 mg/L of VS, respectively. As a result of these processes, it was noted that the use of Caa and VS increases the solubility of Fe at a higher pH.

Mass mortality of the keratose sponge Sarcotragus foetidus in the Aegean Sea (Eastern Mediterranean) correlates with proliferation of Vibrio bacteria in the tissues.

In the last two decades, episodes of mass mortality in benthic communities have often been associated with climatic anomalies, but the ultimate mechanisms through which they lead to death have rarely been identified. This study reports a mass mortality of wild sponges in the Aegean Sea (Turkey, Eastern Mediterranean), which affected the keratose demosponge Sarcotragus foetidus in September 2021. We examined the occurrence of thermo-dependent bacteria of the genus Vibrio in the sponges, identified through 16S rRNA of colonies isolated from sponge tissue in specific culturing media. Six Vibrio sequences were identified from the sponges, three of them being putatively pathogenic (V. fortis, V. owensii, V. gigantis). Importantly, those Vibrios were isolated from only tissues of diseased sponges. In contrast, healthy individuals sampled in both summer and winter led to no Vibrio growth in laboratory cultures. A 50 years record of sea surface temperature (SST) data for the study area reveals a progressive increase in temperature from 1970 to 2021, with values above 24°C from May to September 2021, reaching an absolute historical maximum of 28.9°C in August 2021. We hypothesize that such elevated SST values maintained for several months in 2021 promoted proliferation of pathogenic Vibrio species (thermo-dependent bacteria) in S. foetidus, triggering or aggravating the course of sponge disease. Thus, vibrioisis emerges as one of the putative mechanisms through which global water warming in the Mediterranean Sea translates into sponge mortality. The historical time course of temperature data for the studied area in the Aegean Sea predicts that recurrent waves of elevated SST are likely to occur in the coming summers. If so, recurrent disease may eventually eliminate this abundant sponge from the sublittoral in the midterm, altering the original bathymetric distribution of the species and compromising its ecological role.

Phylomitogenomics bolsters the high-level classification of Demospongiae (phylum Porifera).

Class Demospongiae is the largest in the phylum Porifera (Sponges) and encompasses nearly 8,000 accepted species in three subclasses: Keratosa, Verongimorpha, and Heteroscleromorpha. Subclass Heteroscleromorpha contains ∼90% of demosponge species and is subdivided into 17 orders. The higher level classification of demosponges underwent major revision as the result of nearly three decades of molecular studies. However, because most of the previous molecular work only utilized partial data from a small number of nuclear and mitochondrial (mt) genes, this classification scheme needs to be tested by larger datasets. Here we compiled a mt dataset for 136 demosponge species-including 64 complete or nearly complete and six partial mt-genome sequences determined or assembled for this study-and used it to test phylogenetic relationships among Demospongiae in general and Heteroscleromorpha in particular. We also investigated the phylogenetic position of Myceliospongia araneosa, a highly unusual demosponge without spicules and spongin fibers, currently classified as Demospongiae incertae sedis, for which molecular data were not available. Our results support the previously inferred sister-group relationship between Heteroscleromorpha and Keratosa + Verongimorpha and suggest five main clades within Heteroscleromorpha: Clade C0 composed of order Haplosclerida; Clade C1 composed of Scopalinida, Sphaerocladina, and Spongillida; Clade C2 composed of Axinellida, Biemnida, Bubarida; Clade C3 composed of Tetractinellida; and Clade C4 composed of Agelasida, Clionaida, Desmacellida, Merliida, Suberitida, Poecilosclerida, Polymastiida, and Tethyida. The inferred relationships among these clades were (C0(C1(C2(C3+C4)))). Analysis of molecular data from M. araneosa placed it in the C3 clade as a sister taxon to the highly skeletonized tetractinellids Microscleroderma sp. and Leiodermatium sp. Molecular clock analysis dated divergences among the major clades in Heteroscleromorpha from the Cambrian to the Early Silurian, the origins of most heteroscleromorph orders in the middle Paleozoic, and the most basal splits within these orders around the Paleozoic to Mesozoic transition. Overall, the results of this study are mostly congruent with the accepted classification of Heteroscleromorpha, but add temporal perspective and new resolution to phylogenetic relationships within this subclass.