Sponge Axinella Research Articles

Short macrocyclic peptides in sponge genomes

Sponges (Porifera) contain many peptide-specialized metabolites with potent biological activities and significant roles in shaping marine ecology. It is well established that symbiotic bacteria produce bioactive "sponge" peptides, both on the ribosome (RiPPs) and nonribosomally. Here, we demonstrate that sponges themselves also produce many bioactive macrocyclic peptides, such as phakellistatins and related proline-rich macrocyclic peptides (PRMPs). Using the Stylissa carteri sponge transcriptome, methods were developed to find sequences encoding 46 distinct RiPP-type core peptides, of which ten encoded previously identified PRMP sequences. With this basis set, the genome and transcriptome of the sponge Axinella corrugata was interrogated to find 35 PRMP precursor peptides encoding 31 unique core peptide sequences. At least 11 of these produced cyclic peptides that were present in the sponge and could be characterized by mass spectrometry, including stylissamides A-D and seven previously undescribed compounds. Precursor peptides were encoded in the A. corrugata genome, confirming their animal origin. The peptides contained signal peptide sequences and highly repetitive recognition sequence-core peptide elements with up to 25 PRMP copies in a single precursor. In comparison to sponges without PRMPs, PRMP sponges are incredibly enriched in potentially secreted polypeptides, with >23,000 individual signal peptide encoding genes found in a single transcriptome. The similarities between PRMP biosynthetic genes and neuropeptides in terms of their biosynthetic logic suggest a fundamental biology linked to circular peptides, possibly indicating a widespread and underappreciated diversity of signaling peptide post-translational modifications across the animal kingdom.

Effect of denaturing agents on a lectin isolated from marine sponge Axinella donnaniand its potential cytotoxic activity

Marine sponges are oldest metazoans which were found to have structural and biologically diverse lectins. A lectin isolated from marine sponge Axinella donnani (ADL) were evaluated for its stability against denaturing agents such as Urea, Thiourea and Guanidine hydrochloride. Urea and Thiourea decreases the stability of ADL in higher concentration (5 mM) while Guanidine hydrochloride does not show any activity. Finally, cyt otoxicity of ADL at different concentrations (0.25 500 mg/mL) was tested against MCF 7, A431 and HeLa cell lines by MTT assay. ADL exhibited potential cytotoxicity against these cell lines after 24 hours in a dose dependent manner. Thus, ADL could be a pot ent cytotoxic agent in clinical applications.

Possible Population Growth of Astrospartus mediterraneus (Risso, 1826) (Ophiuroidea, Gorgonocephalidae) in the Mediterranean Sea

During the last decades, the number of observations of the basket star Astrospartus mediterraneus (Risso, 1826) in the Mediterranean Sea has significantly grown, thanks to SCUBA diver and ROV sightings, citizen reports, as well as particularly large catches by the artisanal fishery. Having been generally considered rare in many areas of the basin, such a long-term increase of records might assign to this basket star the putative role of a winner species in the context of climate changes. In the present study, we combined the overall literature information with the data available for the Ligurian Sea collected during extensive ROV campaigns conducted between 2012 and 2022 at a depth ranging from 20 to 123 m, to better understand the distribution and abundance of this species. The basket star was observed in almost the whole explored bathymetric range living on gorgonians (Eunicella cavolini, E. verrucosa, Paramuricea clavata, and Leptogorgia sarmentosa) and massive sponges (Aplysina cavernicola, Sarcotragus foetidus, Spongia lamella, and Axinella polypoides). In the considered period, the number of recorded specimens did not show a clear trend, but differences emerged over years and months. These variations were strongly correlated with rainfall amounts that, in oligotrophic waters, such as those of the Ligurian Sea, represent an important input of organic matter for these passive filter feeders, especially in the summertime.

Axinellamine E, One New Pyrrololactam Alkaloid from the South China Sea Sponge Axinella sp.

Chemical investigation of the EtOH extract of the sponge Axinella sp. collected from the South China Sea resulted in the identification of one new pyrrololactam alkaloid, axinellamine E (2), along with four known analogs (1, 3-5). Compound 1 was initially separated as enantiomers and was further separated to be optically pure compounds (1 a and 1 b) by a chiral column. The planar structure of compound 2 was determined mainly by 1D-, 2D-NMR, and HR-ESI-MS data analyses. Absolute configurations of 1 a and 1 b was defined by calculated ECD spectra method. All of the compounds were evaluated for their anti-inflammatory activities against nitric oxide production in lipopolysaccharide-induced RAW 264.7 cells among which compound 1 showed weak activity at 40 μg/mL. Plausible biosynthetic pathways corresponding to aldisine analogs of 1, 2, 4, and 5 were also discussed.

Axinellasins A-D, Immunosuppressive Cycloheptapeptide Diastereomers, Discovered via a Precursor Ion Scanning-Supercritical Fluid Chromatography Strategy from the Marine Sponge Axinella species.

The precursor ion scanning-supercritical fluid chromatography (PI-SFC) method was applied to explore new methionine sulfoxide-containing cycloheptapeptides, axinellasins A-D (1-4), from the marine sponge Axinella sp. Their structures, including absolute configurations, were elucidated by detailed spectroscopic analyses and X-ray crystallography. The total synthesis of 4 was completed via an Fmoc solid/solution-phase synthesis. Compounds 1-4 exhibited immunosuppressive effects via inhibition of T and B cell proliferation, and 1 and 4 showed better inhibitory activities than their corresponding diastereomers.

10Z‑Hymenialdisine inhibits angiogenesis by suppressing NF‑κB activation in pancreatic cancer cell lines.

10Z-Hymenialdisine is a natural product derived from the marine sponge Axinella carteri. 10Z-Hymenialdisine has anti-inflammatory effects exerted through NF-κB; however, it is unclear whether 10Z-Hymenialdisine has anti-angiogenic effects in cancer cells. In the present study, both the anti-angiogenic and antimetastatic effects of this compound in pancreatic cancer were investigated. It was initially confirmed that 10Z-Hymenialdisine significantly inhibited the proliferation of pancreatic cancer cells. Next, using both reverse transcription-quantitative PCR and ELISA, it was demonstrated that 10Z-Hymenialdisine significantly suppressed the expression of VEGF and IL-8 mRNAs and proteins in pancreatic cancer. Immunohistochemical analysis revealed that 10Z-Hymenialdisine inhibited NF-κB activity in pancreatic cancer cell lines. It was also identified that 10Z-Hymenialdisine inhibited tube formation in EA.hy926 cells. In vivo, 10Z-Hymenialdisine significantly inhibited the growth of BxPC-3 pancreatic cancer cells that were subcutaneously injected into model mice. In conclusion, the present study demonstrated that 10Z-Hymenialdisine exerted anti-angiogenic effects by suppressing NF-κB activity and angiogenic factors, such as VEGF and IL-8, in pancreatic cancer cell lines. 10Z-Hymenialdisine has potential applications as a novel therapeutic agent for the treatment of pancreatic cancer.

Heavy metal accumulation capacity of Axinella damicornis (Esper, 1794) (Porifera, Demospongiae): a tool for bioremediation of polluted seawaters

Within the overwhelming increase of pollutants spilled into the sea, heavy metals are emerging contaminants due to their high toxicity, persistence, and accumulation in both the organisms and the environment. This study aims to assess the retention capability of lead and cadmium by the sponge Axinella damicornis under laboratory conditions. The sponges were exposed for 144 h to seawaters artificially polluted with Pb and Cd separately and with a mixture of the two metals. The retention efficiency of the sponge in removing the metals from seawater and the metal uptake in the sponge body at the end of the experiment were measured. The highest values were recorded for Pb: in the sponge, this metal resulted in 6 times and 9 times more concentrated than Cd, in the case of single and double contamination respectively. The metal concentrations, especially for Pb, were much higher in A. damicornis than in other organisms investigated in the sea. The synergistic effects of the mixture of Pb and Cd were also evaluated, and remarkable signs of stress and necrosis were recorded in the specimens exposed to the two metals combined. This study paves the way to the knowledge increasing on both the effective effects of heavy metal contamination on the organisms and on the possible use of A. damicornis as efficient tool for bioremediation of polluted seawaters.

10Z-Hymenialdisine Inhibits Angiogenesis by Suppressing NF-κB Activation in Pancreatic Cancer Cell Lines

Introduction: 10Z-Himenialdicin is a natural product derived from the marine sponge Axinella carteri. It has reported that 10Z-Himenialdicin has an anti-angiogenetic effect through the NF-κB which produces angiogenic factors such as VEGF and IL-8. Although there is no previous report which revealed the anti-angiogenetic effects of 10Z-Himenialdicin on cancer cells, there is a potential of inhibiting the anti-angiogenetic and prevent the metastasis in pancreatic cancer.

A Multi-Species Investigation of Sponges' Filtering Activity towards Marine Microalgae.

Chronic discharge of surplus organic matter is a typical side effect of fish aquaculture, occasionally leading to coastal eutrophication and excessive phytoplankton growth. Owing to their innate filter-feeding capacity, marine sponges could mitigate environmental impact under integrated multitrophic aquaculture (IMTA) scenarios. Herein, we investigated the clearance capacity of four ubiquitous Mediterranean sponges (Agelas oroides, Axinella cannabina, Chondrosia reniformis and Sarcotragus foetidus) against three microalgal substrates with different size/motility characteristics: the nanophytoplankton Nannochloropsis sp. (~3.2 μm, nonmotile) and Isochrysis sp. (~3.8 μm, motile), as well as the diatom Phaeodactylum tricornutum (~21.7 μm, nonmotile). In vitro cleaning experiments were conducted using sponge explants in 1 L of natural seawater and applying different microalgal cell concentrations under light/dark conditions. The investigated sponges exhibited a wide range of retention efficiencies for the different phytoplankton cells, with the lowest average values found for A. cannabina (37%) and the highest for A. oroides (70%). The latter could filter up to 14.1 mL seawater per hour and gram of sponge wet weight, by retaining 100% of Isochrysis at a density of 105 cells mL−1, under darkness. Our results highlight differences in filtering capacity among sponge species and preferences for microalgal substrates with distinct size and motility traits.

Cultivation of fractionated cells from a bioactive-alkaloid-bearing marine sponge Axinella sp.

Sponges are among the most primitive multicellular organisms and well-known as a major source of marine natural products. Cultivation of sponge cells has long been an attractive topic due to the prominent evolutionary and cytological significance of sponges and as a potential approach to supply sponge-derived compounds. Sponge cell culture is carried out through culturing organized cell aggregates called 'primmorphs.' Most research culturing sponge cells has used unfractionated cells to develop primmorphs. In the current study, a tropical marine sponge Axinella sp., which contains the bioactive alkaloids, debromohymenialdisine (DBH), and hymenialdisine (HD), was used to obtain fractionated cells and the corresponding primmorphs. These alkaloids, DBH and HD, reportedly show pharmacological activities for treating osteoarthritis and Alzheimer's disease. Three different cell fractions were obtained, including enriched spherulous cells, large mesohyl cells, and small epithelial cells. These cell fractions were cultivated separately, forming aggregates that later developed into different kinds of primmorphs. The three kinds of primmorphs obtained were compared as regards to appearance, morphogenesis, and cellular composition. Additionally, the amount of alkaloid in the primmorphs-culture system was examined over a 30-d culturing period. During the culturing of enriched spherulous cells and developed primmorphs, the total amount of alkaloid declined notably. In addition, the speculation of alkaloid secretion and some phenomena thatoccurred during cell culturing are discussed.

Isolation, identification, and HPTLC quantification of dehydrodeoxycholic acid from Persian Gulf sponges

This study aims to investigate the chemical constituents of sponges Dysidea avara (D. avara) and Axinella sinoxea (A. sinoxea), grown up in the Persian Gulf, as well as dehydrodeoxycholic acid (DHCA) content in methanolic extracts of the selected sponges. The chromatography-mass spectrometry (GC–MS) fingerprint of bioactive compounds from methanolic extracts of the selected marine sponge samples was investigated. Based on molecular docking results, among chemical compounds found in marine sponges, DHCA has anti-inflammatory and antipsoriatic properties. They also indicated that DHCA generated stable complexes with 1w81, 3bqm, and 3k8o receptors (psoriasis-related targets) with a binding energy (BE) of −9.26, −10.62, and −7.59 kcal mol−1, respectively. DHCA is isolated from the methanolic extracts of marine sponge samples on chromatographic plates was quantified after derivatization with anisaldehyde reagent by the validated HPTLC method. In-situ HPTLC-DPPH was also calculated to evaluate the free radical-scavenging activity (FRSA) of DHCA. In-silico ADME (Absorption, Distribution, Metabolism, Excretion) predictions revealed that the compound had minimum toxicity and acceptable human intestinal absorption (HIA), as well as low skin permeability. These can potentially be employed as lead compounds to develop a novel antipsoriatic drug.

Recifin A, Initial Example of the Tyr-Lock Peptide Structural Family, Is a Selective Allosteric Inhibitor of Tyrosyl-DNA Phosphodiesterase I.

Tyrosyl-DNA phosphodiesterase 1 (TDP1) is a molecular target for the sensitization of cancer cells to the FDA-approved topoisomerase inhibitors topotecan and irinotecan. High-throughput screening of natural product extract and fraction libraries for inhibitors of TDP1 activity resulted in the discovery of a new class of knotted cyclic peptides from the marine sponge Axinella sp. Bioassay-guided fractionation of the source extract resulted in the isolation of the active component which was determined to be an unprecedented 42-residue cysteine-rich peptide named recifin A. The native NMR structure revealed a novel fold comprising a four strand antiparallel β-sheet and two helical turns stabilized by a complex disulfide bond network that creates an embedded ring around one of the strands. The resulting structure, which we have termed the Tyr-lock peptide family, is stabilized by a tyrosine residue locked into three-dimensional space. Recifin A inhibited the cleavage of phosphodiester bonds by TDP1 in a FRET assay with an IC50 of 190 nM. Enzyme kinetics studies revealed that recifin A can specifically modulate the enzymatic activity of full-length TDP1 while not affecting the activity of a truncated catalytic domain of TDP1 lacking the N-terminal regulatory domain (Δ1-147), suggesting an allosteric binding site for recifin A on the regulatory domain of TDP1. Recifin A represents both the first of a unique structural class of knotted disulfide-rich peptides and defines a previously unseen mechanism of TDP1 inhibition that could be productively exploited for potential anticancer applications.

Marine Fungus Aspergillus chevalieri TM2-S6 Extract Protects Skin Fibroblasts from Oxidative Stress.

The strain Aspergillus chevalieri TM2-S6 was isolated from the sponge Axinella and identified according to internal transcribed spacer (ITS) molecular sequence homology with Aspergillus species from the section Restricti. The strain was cultivated 9 days on potato dextrose broth (PDB), and the medium evaluated as antioxidant on primary normal human dermal fibroblasts (NHDF). The cultivation broth was submitted to sterile filtration, lyophilized and used without any further processing to give the Aspergillus chevalieri TM2-S6 cultivation broth ingredient named ACBB. ACCB contains two main compounds: tetrahydroauroglaucin and flavoglaucin. Under oxidative stress, ACCB showed a significant promotion of cell viability. To elucidate the mechanism of action, the impact on a panel of hundreds of genes involved in fibroblast physiology was evaluated. Thus, ACCB stimulates cell proliferation (VEGFA, TGFB3), antioxidant response (GPX1, SOD1, NRF2), and extracellular matrix organization (COL1A1, COL3A1, CD44, MMP14). ACCD also reduced aging (SIRT1, SIRT2, FOXO3). These findings indicate that Aspergillus chevalieri TM2-S6 cultivation broth exhibits significant in vitro skin protection of human fibroblasts under oxidative stress, making it a potential cosmetic ingredient.

Integrating Molecular Networking and 1H NMR Spectroscopy for Isolation of Bioactive Metabolites from the Persian Gulf Sponge Axinella sinoxea.

The geographic position, highly fluctuating sea temperatures and hypersalinity make Persian Gulf an extreme environment. Although this unique environment has high biodiversity dominated by invertebrates, its potential in marine biodiscovery has largely remained untapped. Herein, we aimed at a detailed analysis of the metabolome and bioactivity profiles of the marine sponge Axinella sinoxea collected from the northeast coast of the Persian Gulf in Iran. The crude extract and its Kupchan subextracts were tested in multiple in-house bioassays, and the crude extract and its CHCl3-soluble portion showed in vitro antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecium (Efm). A molecular networking (MN)-based dereplication strategy by UPLC-MS/MS revealed the presence of phospholipids and steroids, while 1H NMR spectroscopy indicated the presence of additional metabolites, such as diketopiperazines (DKPs). Integrated MN and 1H NMR analyses on both the crude and CHCl3 extracts combined with an antibacterial activity-guided isolation approach afforded eight metabolites: a new diketopiperazine, (-)-cyclo(L-trans-Hyp-L-Ile) (8); a known diketopiperazine, cyclo(L-trans-Hyp-L-Phe) (7); two known phospholipids, 1-O-hexadecyl-sn-glycero-3-phosphocholine (1) and 1-O-octadecanoyl-sn-glycero-3-phosphocholine (2); two known steroids, 3β-hydroxycholest-5-ene-7,24-dione (3) and (22E)-3β-hydroxycholesta-5,22-diene-7,24-dione (4); two known monoterpenes, loliolide (5) and 5-epi-loliolide (6). The chemical structures of the isolates were elucidated by a combination of NMR spectroscopy, HRMS and [α]D analyses. All compounds were tested against MRSA and Efm, and compound 3 showed moderate antibacterial activity against MRSA (IC50 value 70 μg/mL). This is the first study that has dealt with chemical and bioactivity profiling of A. sinoxea leading to isolation and characterization of pure sponge metabolites.

Cloning and expression of ATP N-glycosidase from the freshwater sponge Ephydatia muelleri

Previously we had discovered unusual enzymatic activity in the marine sponge Axinella polypoides, ATP N-glycosidase (Reintamm et al., 2003). We show here that the Ephydatia muelleri mRNA encoding protein with PNP_UDP_1 (phosphorylase superfamily) signature is the secreted ATP N-glycosidase. The functionality of the protein was established by recombinant expression in Pichia pastoris. In addition to the enzymatic domain, the full-length protein contains the N-terminal cysteine-rich domain belonging to the subfamily SCP_HrTT-1 (cd05559) of the SCP (sperm coating protein) superfamily (cl00133).

Fungal diversity of mangrove-associated sponges from New Washington, Aklan, Philippines

ABSTRACTSponge-associated fungi are the least explored marine fungal groups. It is only in recent years that fungal symbionts of marine sponges have received attention mainly due to the isolation of bioactive metabolites while not much attention was given to their specificity, biogeography and exact roles in marine sponges. The diversity of fungi associated with mangrove sponges (Axinella sp., Halichondria cf. panicea, Haliclona sp., Tedania sp.) collected from New Washington, Aklan, Philippines were investigated using morphological observation. A total of 110 species of sponge-associated fungi belonging to 22 genera of ascomycetes with 18 genera of asexual morphs whose sexual stage is unknown, 2 genera of basidiomycetes, 21 morphospecies of Mycelia sterilia, 1 unidentified yeast species and 11 unidentified hyphomycetes were isolated from four species of mangrove sponges. This is the first study that explored the diversity and ecology of sponge-associated fungi in mangrove habitats from the Philippines. The results of the study suggest host-preference by various fungal taxa and the development of fungi on these hosts appeared to be strongly influenced by the characteristics or nature of the immediate environment.

The park never born: Outcome of a quarter of a century of inaction on the sea‐floor integrity of a proposed but not established Marine Protected Area

Abstract Marine Protected Areas (MPAs) are a major tool to conserve marine ecosystems but are also strongly attractive to tourists, the increased numbers of which can cause environmental issues if not properly managed. Proposing an MPA and then failing to establish it risks advertising the beauty of a marine area without managing the unavoidable increase in tourism. This is what happened at Gallinara Island (Ligurian Sea, north‐western Mediterranean), where an MPA was proposed in 1990 but has not yet been established. Benthic data collected soon after the proposal (1990–92) have been compared with similar information obtained in subsequent periods (2009 and 2016): change has been assessed both visually and statistically, using multivariate and univariate techniques. The Gallinara sea floor supported five epibenthic communities in 1991, four of which corresponded to European Nature Information System (EUNIS) habitats; the fifth epibenthic community, characterized by the erect sponge Axinella polypoides, should be included in the next revision of EUNIS. In the 25 years since the original MPA proposal, all epibenthic communities have exhibited a reduction in their three‐dimensional structure and biotic homogenization, and severe decreases in species diversity, and the sea floor has been littered with derelict fishing gear and other waste from boats. In absence of the MPA, proposals to develop a management plan for a marine Site of Community Importance (SCI) have been provided, to reduce sea‐based human pressures. The dramatic alteration of Gallinara sea floor after 25 years of negligence and inaction is a sad warning for other unmanaged yet beautiful natural areas within urbanized regions of the world.

Two new pyrrolo-2-aminoimidazoles from a Myanmarese marine sponge, Clathria prolifera.

Marine organisms such as marine sponges and soft corals are valuable sources of pharmacologically active secondary metabolites. In our ongoing research on the discovery of new secondary metabolites from marine organisms, two new pyrrolo-2-aminoimidazoles, clathriroles A (1) and B (2), were isolated from the water-soluble portion prepared from the methanol and acetone (2:1) extract of the marine sponge, Clathria prolifera, collected in Myanmar. The chemical structures of the isolated compounds were determined using extensive spectroscopic techniques, including NMR, HRESIMS, IR, and optical rotation, and comparisons with the reported literature. The spectroscopic analyses of 1 and 2 suggested that 1 is an enantiomer of antifungal N-methylmanzacidin C isolated from the marine sponge Axinella brevistyla, whereas 2 is a diastereomer of manzacidin D at C-11 isolated from the marine sponge Astrosclera willeyana. To the best of our knowledge, this is the first report of the isolation of the pyrrolo-2-aminoimidazole compounds from C. prolifera. Furthermore, in contrast to the potency of N-methylmanzacidin C against Saccharomyces cerevisiae, the antifungal assay revealed that 1 and 2 lack any activity against this strain. Thus, these observations may suggest that the absolute configurations at both C-9 and C-11 play an important role in controlling the antifungal activity of this type of compound.

Antibacterial Activity Symbiotic Fungi of Marine Sponge Axinella sp., Aspergillus Sydowii on Four Growth Medium

Many infectious diseases caused by Escherichia coli and Staphylococcus aureus which turned into a resistant pathogen. A symbiotic fungi of marine sponge Axinella sp., Aspergillus sydowii from the waters of Riung, East Nusa Tenggara, Indonesia showed antibacterial activity, cultured on the four media, MEB (ST), Noni Juice Media (MG), avocado leaves media (AL), and Soursop leaves media (SR). The symbiotic fungi was cultured for 14 days on each media. The largest weight of symbiotic fungi biomass on ST media 138,95gr and at least 99,12gr of AL media. Purification of bioactive compound is carried out using separatory funnel, and column chromatography. The highest rendemen of extracts on SR media was 3,67%, while the lowest in ST media was 1,22%. The bioactive test used diffusion agar method. Fungi extracts from four mediums have bioactivity against, E. coli and S. aureus. The biggest inhibition zone obtained from the extract of MG KN-15-3-1-3, with inhibition zone 10.71mm and 10.98mm against E. coli and S. aureus.

Lactones from the Sponge-Derived Fungus Talaromyces rugulosus.

The marine-derived fungus Talaromyces rugulosus isolated from the Mediterranean sponge Axinella cannabina and cultured on solid rice medium yielded seventeen lactone derivatives including five butenolides (1–5), seven (3S)-resorcylide derivatives (6–12), two butenolide-resorcylide dimers (13 and 14), and three dihydroisocoumarins (15–17). Among them, fourteen compounds (1–3, 6–16) are new natural products. The structures of the isolated compounds were elucidated by 1D and 2D NMR (Nuclear Magnetic Resonance) spectroscopy as well as by ESI-HRMS (ElectroSpray Ionization-High Resolution Mass Spectrometry). TDDFT-ECD (Time-Dependent Density Functional Theory-Electronic Circular Dichroism) calculations were performed to determine the absolute configurations of chiral compounds. The butenolide-resorcylide dimers talarodilactones A and B (13 and 14) exhibited potent cytotoxicity against the L5178Y murine lymphoma cell line with IC50 values of 3.9 and 1.3 µM, respectively.