Natural products potential of Dictyoceratida sponges-associated micro-organisms

The marine environment represents one of the most underexplored environments in the world. Marine sponges have a higher taxonomic diversity according to definite environmental conditions. They have been considered interesting sources for bioactive compounds. Dictyoceratida sponges are divided into five families which are widely distributed and habituating different types of micro-organisms. However, some secondary metabolites are probably not produced by the sponges themselves, but rather by their associated micro-organisms. These secondary metabolites are characterized by different chemical structures and consequently different biological activities. This review outlines the reported secondary metabolites from micro-organisms associated with Dictyoceratida sponges and their investigated biological activities from 1991 to 2019. The increasing research studies in this field can play a major role in marine microbial natural products drug discovery in the future.     

Molecular phylogeny of the freshwater sponges in Lake Baikal

The phylogenetic relationship of the freshwater sponges (Porifera) in Lake Baikal is not well understood. A polyphyletic and/or monophyletic origin have been proposed. The (endemic) Baikalian sponges have been subdivided into two families: endemic Lubomirskiidae and cosmopolitan Spongillidae. In the present study, two new approaches have been made to resolve the phylogenetic relationship of Baikalian sponges; analysis of (1) nucleotide sequences from one mitochondrial gene, the cytochrome oxidase subunit I (COI) and of (2) one selected intron from the tubulin gene. Specimens from the following endemic Baikalian sponge species have been studied; Lubomirskia baicalensis , Baikalospongia intermedia, Baikalospongia recta , Baikalospongia bacillifera and Swartschewskia papyracea . They are all grouped to the family of Lubomirskiidae. Sequence comparisons were performed with the ubiquitously distributed freshwater sponge Spongilla lacustris (family Spongillidae) as well as with one marine sponge, Suberites domuncula . A sequence comparison * * The sequences reported here are being deposited in the EMBL data base. of the mitochondrial COI gene revealed a monophyletic grouping of the endemic Baikalian sponges with S. lacustris as the most related species to the common ancestor. The sequences of the COI gene from B. recta , B. intermedia , B. bacillifera and L. baicalensis were found to be identical and separated from those of S. lacustris and S. papyracea . In a second approach, the exon/intron sequences framing the intron‐2 of the sponge tubulin gene were chosen for the phylogenetic analysis. The intron sequences were aligned and used for construction of a phylogenetic tree. This analysis revealed again a monophyletic grouping with S. lacustris as the closest related species to the common ancestor. It is concluded that the Baikalian sponges, which have been studied here, are of monophyletic origin. Furthermore, the data suggest that the endemic species S. papyracea is the phylogenetically oldest, extant, endemic Baikalian sponge species.     

Cloning of Hsp70 genes from the marine sponges Sycon raphanus (Calcarea) and Rhabdocalyptus dawsoni (Hexactinellida). An approach to solve the phylogeny of sponges

The phylogenetic relationships among the three classes of the Porifera-Demospongiae, Calcarea and Hexactinellida-are still unresolved, despite the use of molecular analyses of rRNA. To determine whether phylogenetic resolution of these classes is possible based on genes coding for specific proteins, in the present study the genes for the 70 kDa heat shock protein [Hsp70] were isolated from Rhabdocalyptus dawsoni [Hexactinellida] and from Sycon raphanus [Calcarea], and compared to that previously isolated from the demosponge Geodia cydonium. The gene from R. dawsoni is 2021 bp long and encodes a predicted Hsp70 of Mr 77, 697; the protein comprises the characteristic sites of eukaryotic, cytoplasmic Hsp70 polypeptides. The Hsp70 isolated from cDNA from S. raphanus is 2326 bp long. It encodes a potential polypeptide of Mr 85, 927 and belongs to the same class of Hsp70s. All three sponge sequences for Hsp70 were found to be highly identical to both human and plant Hsp70s. The degree of identity at the amino acid (aa) level between the sponge sequences and the human sequence for Hsp70 is 77%-84% and at the nucleotide (nt) level, between 69% and 75%. Resolution of the phylogenetic relationship between the three classes of sponges based on the Hsp70 was not possible due to the high degree of identity [similarity] of their respective aa sequences, which ranged from 80% [90%] to 82% [91%]. The evolutionary rates-K_aa-values-calculated for the sponge Hsp70 molecules, are low, reflecting the strong functional contraints placed upon these polypeptides. These values range from 0.125 times 10^-9 for G. cydonium and R. dawsoni to 0.087 times 10^-9 for S. raphanus. Higher values have previously been reported for the G. cydonium galectin molecule [K_aa-value of 1.7 times 10^-9] and the receptor tyrosine kinase [1.24 times 10^-9] from the same animal. The occurrence of at least one double mutation, in the codon for the aa Ser in the conserved regions of the Hsp70 sequences, also suggests that these molecules are subjected to strong functional constraints.     

Distribution of sponges on the Mauritanian continental shelf

Transect sampling on the continental shelf off the Banc d'Arguin, Mauritania, revealed a patchy distribution of sponge species and populations. Major sponge habitats are inshore coastal sandstone flats harbouring a diverse infaunal sponge fauna, and the moving substrate of shells of live gastropods or empty shells occupied by hermit crabs which harbour a numerically important but species-poor incrusting sponge fauna. The latter forms occur over most of the depth profile. Horizontal distribution of the sponges shows a distinct gap in species and numbers associated with a muddy bottom area where the sediment-rich Banc d'Arguin run-off is found. North and south of this gap the specific composition of the infaunal sponges is considerably different and this is explained as evidence for limitation of range extensions, notably of the southern sponge fauna, by the muddy area.     

Glass sponges (Porifera, Hexactinellida) of the northern Mid-Atlantic Ridge

Glass sponges (Hexactinellida) collected under the framework of the MAR-ECO project on the G.O. Sars cruise to the northern Mid-Atlantic Ridge, between the Azores and the Reykjanes Ridge, and on the 49th cruise of Akademik Mstislav Keldysh to the Charlie-Gibbs Fracture Zone are described. Fourteen species were identified in the material. This relatively rich fauna includes several novel findings, indicating that the hexactinellid fauna of the northern Mid-Atlantic Ridge is poorly investigated. One genus, Dictyaulus, has never before been reported from the Atlantic Ocean. Two species belonging to the genera Heterotella and Amphidiscella are new to science. Two other species, Rossella nodastrella Topsent, 1915 and Doconestes sessilis Topsent, 1928, have been collected just one other time. Finally, a probable new genus of Euplectellidae, which unfortunately cannot be adequately described because of how small the specimens are, is represented among these collections. A large portion of dead, rigid skeletons of Euretidae and spicule mats of Rossellidae are also reported but not described in detail. Representation of some genera in the Charlie-Gibbs Fracture Zone, coupled with recent observations from elsewhere along the Mid-Atlantic mountain chain, suggests that this fauna is as similar to those of the Indian Ocean and Indo-West Pacific as it is to West or East Atlantic faunas.     

Phototactic responses of larvae from the marine sponges Neopetrosia proxima and Xestospongia bocatorensis (Haplosclerida: Petrosiidae)

Abstract. Previous studies suggest that phototaxis in sponge larvae is generated by the bending of a tuft of long posterior cilia (LPC). The photoresponsiveness of these cilia is often assayed by examining their reaction to sudden changes in light intensity. Here, we document and describe the larvae of the tropical marine sponges Neopetrosia proxima and Xestospongia bocatorensis and examine the phototactic behavior of their larvae. Both species brood ovoid, tufted parenchymella larvae, clearly countering an earlier hypothesis that all petrosid sponges are oviparous. Larvae of N. proxima were positively phototactic and settled after 2 d, while larvae of X. bocatorensis were negatively phototactic and settled in as little as 4 h. In both species, LPC quickly responded to changes in the light intensity. When the light intensity is reduced, the larvae of N. proxima fold the cilia inwards immediately without beating, then flare them outwards, beating for a few seconds, and then gradually return to the neutral position while continuing to beat. In contrast, the larvae of X. bocatorensis flare the cilia outwards when the light intensity is reduced and fold them inwards when the light intensity is increased. Comparisons with reported ciliary responses to light for other species demonstrate that these responses do not show the hypothesized one-to-one correspondence with phototactic behaviors and are, therefore, of limited use in explaining the mechanisms that coordinate larval swimming.     

Spicule dimensions as taxonomic criteria in the identification of haplosclerid sponges from the shores of Anglesey

The lengths and widths of at least 100 spicules from each of 126 specimens, comprising at least 12 species of haplosclerid sponges, mainly from Church Island and Rhosneigr, Anglesey, North Wales have been measured. It was found that spicule dimensions by themselves would be unreliable in the identification of species. The sample means, medians and maxima, when plotted using width and length axes, form specific clusters that overlap, sometimes to a considerable extent. The correlation coefficients for width against length varied widely, even when single species were considered. The variation arises from differences in the numbers of juvenile spicules in various samples. The best correlation coefficients exceeded 0.8 and the regression constants for linear correlation in such samples could be useful in the diagnosis and taxonomy of the species. The coefficients of variation for length and width also yielded parameters for species characterization. In general the coefficient for length tended to increase as the mean length increased, whereas that for width tended to decrease with increasing mean width. Some species stood apart from the general trends, however. The coefficients varied widely from one sample to another of the same species, identified using a variety of diagnostic features. To some extent the variation was linked with the date of collection. When all the data were combined, the average coefficient for width decreased markedly in May. The same was true for two species that were separately considered. The decrease was not simply caused by a change in mean width, changes in standard deviation also being involved. There was a tendency for certain species, in particular Reniera rosea and Gellius angulatus , to produce thin spicules in the spring, which became incorporated distally in the primary spicule bundles in July-August.     

Histological investigation of organisms with hard skeletons: a case study of siliceous sponges

Siliceous and calcareous sponges commonly are treated with acid to remove the spicules prior to embedding and cutting for histological investigations. Histology of spiculated sponge tissue represents a challenging problem in sponge histotechnology. Furthermore, fluorescence in situ hybridization (FISH), a key method for studying sponge-associated microbes, is not possible after acid treatment. For a broad range of siliceous sponge species, we developed and evaluated methods for embedding in paraffin, methylmethacrylate resins, LR White resin and cryomatrix. Different methods for cutting tissue blocks as well as mounting and staining sections also were tested. Our aim was to enable histological investigations and FISH without prior removal of the spicules. To obtain an overview of tissue and skeleton arrangement, we recommend embedding tissue blocks with LR White resin combined with en bloc staining techniques for large specimens with thick and numerous spicules, but paraffin embedding and subsequent staining for whole small specimens. For FISH on siliceous sponges, we recommend Histocryl embedding if the spicule content is high, but paraffin embedding if it is low. Classical histological techniques are used for detailed tissue examinations.     

The extent of asexual reproduction in sponges of the genus Chondrilla (Demospongiae: Chondrosida) from the Caribbean and the Brazilian coasts

In this study, we estimated the extent of asexual reproduction and genotypic diversity at the intra- and interspecific levels in sponges of the genus Chondrilla in the Caribbean and along the coast of Brazil. Allozymes were used to identify the genotypes of specimens of Chondrilla in each location. The four species studied displayed a large variation in the extent of clonal reproduction and genotypic diversity, with the two species from the Bahamas having a greater proportion of asexually produced individuals than those along the coast of Brazil. Conspecific Brazilian populations of Chondrilla sp. had large differences in clonality: the population from a heterogeneous environment and under the influence of a strong upwelling had little clonality (7%), whereas the population located 350 km south along the coast, in a more homogeneous and temporally stable environment, had a five-fold larger (39%) proportion of asexually derived individuals. Finally, we were able to confirm that, besides fission, the genus Chondrilla displays a second mode of asexual reproduction, by fragmentation of teardrop shaped individuals.     

Growth patterns of Lower Palaeozoic sponges

Detailed studies of the growth patterns of modern siliceous sponges are restricted to demosponges and theoretical models. It is generally assumed that sponge growth is essentially incremental, with completion of one arbitrary unit being followed by external addition. All recent species are thick-walled, but Lower Palaeozoic sponges are dominated by thin-walled hexactinellids, with most Cambrian taxa consisting of a single spicule layer. Large populations of a primitive dictyospongiid have allowed the reconstruction of the growth patterns of their spicules and body morphology. The results indicate that growth occurred through continuous expansion of the globose body, accompanied by continuous enlargement of existing spicules, with a spicule size limit being reached only during the lifetime of a few individuals. It is noted that this skeletal growth pattern is otherwise restricted to deuterostomes. Consecutive appearance of successive spicule size orders appears to have maintained a maximum inhalant pore area. Comparisons with more limited data from two acanthose hexactinellids and a hazeliid demosponge indicate that an identical growth pattern operated in these species. The subsequent evolution of growth patterns is discussed, with various mechanisms producing the later thick-walled morphologies of hexactinellids and demosponges. The implications of these observations are discussed with reference to identification and systematics, since spicule size and arrangement are shown to vary during growth.     

Ubiquitins (polyubiquitin and ubiquitin extension protein) in marine sponges: cDNA sequence and phylogenetic analysis

The complete nucleotide sequences of two Suberites domuncula cDNAs and one Sycon raphanus cDNA, all encoding ubiquitin, have been determined. One cDNA from S. domuncula codes for polyubiquitin with four tandemly repeated monomeric units and the second cDNA encodes ubiquitin fused to a ribosomal protein of 78 amino acids (aa). S. domuncula possesses at least one additional polyubiquitin gene, from which the last two monomers were also sequenced. All analysed genes from S. domuncula encode identical ubiquitin proteins, with only one aa difference (Ala 19) to the human/higher animals ubiquitin (Pro 19). Ubiquitin in S. domuncula is identical with the ubiquitin found in another Demospongia, Geodia cydonium. The cDNA from S. raphanus encodes polyubiquitin with seven tandemly repeated units. All these gene monomers code for the same ubiquitin, which differs from the human/higher animals ubiquitin only at position 24 (Asp in Sycon, Glu in others). However, ubiquitin from S. raphanus (Calcarea) shows two aa differences (positions 19 and 24), when compared with the ubiquitin sequences from the two Demospongiae. In a phylogenetic tree constructed by multiple sequence alignment of all sponge ubiquitin gene monomers so far identified, all monomers from the same species cluster together, with the clear exception of the monomer from S. domuncula ribosomal protein fusion gene. This monomer branches off first from the tree and forms a separate line; this gives evidence for a very ancient split of ubiquitin-ribosomal-protein fusion genes from polyubiquitin encoding genes and their long separate coexistence in eukaryotes. The ubiquitin extension protein from S. domuncula is 78 aa long, displays all characteristics of 76–81 aa long ribosomal fusion proteins and shows 78% identity in the first 73 aa with the human S27a protein. However, its C-terminal sequence: 69-GLTYVYKKSD-78 is more similar to the plant consensus (69-GLTYVYQ/NK-76), than to the higher animal consensus (69-CLTYCFNK-76). This protein isolated from a sponge, belonging to the phylogenetically oldest multicellular animals, the Porifera, branches off first from the phylogenetic tree of metazoan ubiquitin extension proteins of the small ribosomal subunits.     

Reproductive cycles of the sympatric excavating sponges Cliona celata and Cliona viridis in the Mediterranean Sea

Abstract. Clionaids are excavating sponges, which live in and grow into calcareous substrates. We studied the sexual reproductive cycles of two clionaid sponges coexisting in a Mediterranean coastal basin (Porto Cesareo, Italy), Cliona viridis and Cliona celata, by analyzing monthly tissue samples of ten specimens of each species collected over a 2‐year period. From May to June of the second study year, supplementary samples were taken weekly. Up to 90% of the specimens of C. viridis and 70% of those of C. celata sampled were reproductive during the study. In both species, but particularly in C. viridis, reproductive investment, measured as the percentage of sponge tissue occupied by gametes, was high. Oocytes were present almost year‐round in both species, except for a 1–4‐month period after zygote release. In contrast, spermatogenesis occurred most frequently in May in both species, when (May–June) oocytes reached their greatest diameters. Cliona viridis and C. celata are hermaphrodites, with oocytes and spermatic cysts coexisting in 10% of the studied individuals in the first year of the study, and in 30% during the second. No developing embryos or larvae were incubated in the sponge tissues, and fertilization was not observed. Temperature may play a role in triggering some important phases of the reproduction of these Cliona, such as oocyte maturation and spermatogenesis, which occurred when water temperature increased from 17°C to 25°C between May and June.     

Phylogenetic signal in the evolution of body colour and spicule skeleton in calcareous sponges

Some of the morphological characters used in Porifera taxonomy have often been shown to be inconsistent. In the present study, we tested the phylogenetic coherence of currently used taxonomic characters of the calcarean genus Clathrina. For this, 20 species of Clathrina and three other calcinean genera (Ascandra, Guancha, and Leucetta) were sequenced for the ITS and D2 region of the 28S ribosomal DNA. Maximum‐likelihood and maximum‐parsimony algorithms were used to reconstruct phylogenetic trees. Deep divergences were observed in our tree and Clathrina was shown to be paraphyletic. The major split in our topology showed a clear‐cut distinction between sponges with and without tetractine spicules. Moreover, a group of yellow‐coloured Clathrina was clearly separated from the remaining white‐coloured species. Our results show that the presence of diactines, water‐collecting tubes, the degree of cormus anastomosis, and actine shapes do not correlate with the major clades of the calcinean phylogeny. On the other hand, the presence of tripods, the absence of tetractines, and the presence of spines in the apical actine of tetractines seem to be good synapomorphies for clades in our tree. Our results demonstrate that skeleton characters can be reliably used in higher level taxonomy in Clathrinida. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 163 , 1026–1034.     

Giving the early fossil record of sponges a squeeze

Twenty candidate fossils with claim to be the oldest representative of the Phylum Porifera have been re‐analysed. Three criteria are used to assess each candidate: (i) the diagnostic criteria needed to categorize sponges in the fossil record; (ii) the presence, or absence, of such diagnostic features in the putative poriferan fossils; and (iii) the age constraints for the candidate fossils. All three criteria are critical to the correct interpretation of any fossil and its placement within an evolutionary context. Our analysis shows that no Precambrian fossil candidate yet satisfies all three of these criteria to be a reliable sponge fossil. The oldest widely accepted candidate, Mongolian silica hexacts from c. 545 million years ago (Ma), are here shown to be cruciform arsenopyrite crystals. The oldest reliable sponge remains are siliceous spicules from the basal Cambrian (Protohertzina anabarica Zone) Soltanieh Formation, Iran, which are described and analysed here in detail for the first time. Extensive archaeocyathan sponge reefs emerge and radiate as late as the middle of the Fortunian Stage of the Cambrian and demonstrate a gradual assembly of their skeletal structure through this time coincident with the evolution of other metazoan groups. Since the Porifera are basal in the Metazoa, their presence within the late Proterozoic has been widely anticipated. Molecular clock calibration for the earliest Porifera and Metazoa should now be based on the Iranian hexactinellid material dated to c. 535 Ma. The earliest convincing fossil sponge remains appeared at around the time of the Precambrian‐Cambrian boundary, associated with the great radiation events of that interval.     

Intragenomic variation of the rDNA internal transcribed spacers in sponges (Phylum Porifera): implications for phylogenetic studies

The internal transcribed spacer regions (ITS1 and ITS2) of the tandemly repeated nuclear ribosomal DNA clusters are frequently used as markers for fine scale analyses in diverse animals. In certain taxa, ITS is nearly exclusively used for population level or inter-specific studies, despite the frequent presence of divergent paralogs within individual genomes that can be phylogenetically misleading. For the first time we survey diverse marine sponges to determine the extent and phylogenetic implications of intragenomic polymorphisms (IGPs) exhibited at their ITS loci. We discover that the extent of IGP varies greatly between taxa (with most taxa exhibiting very few) and cannot be predicted by taxonomy. Furthermore, we demonstrate that ITS can be phylogenetically informative between species when moderate levels of IGPs are detected, but that ITS paralogy can interfere with population level studies. We caution against the routine use of ITS in phylogenetic studies of sponges without (1) screening for IGPs in specimens from every population sampled; (2) including all divergent paralogs in phylogenetic analyses; (3) testing ITS data using other single-copy, unlinked loci (such as nuclear introns).     

First evidences of surfactant biodegradation by marine sponges (Porifera): an experimental study with a linear alkylbenzenesulfonate

The main objective of this work was to better appreciate the role of benthic macro-organisms in the biodegradation of detergents in the marine environment. Sponges, which could be highly resistant to pollution and which are highly active filter-feeders, appear as interesting organisms in this topic. An experimental study conducted in aquarium with seawater enriched in a pure linear alkylbenzenesulfonate (LAS), namely 1-(p-Sulfophenyl)nonane, has shown that the primary degradation was 10 times more rapid in the presence of the sponge Spongia officinalis than in the presence of only marine bacteria. The main degradation metabolites, 3-(p-sulfophenyl)propionic acid and p-sulfobenzoic acid were produced in greater amounts in the presence of sponge. The very rapid degradation kinetics observed in this study may be due to the symbiotic microbial community present in S. officinalis. The sponge may also have promoted the activity of marine bacteria by transforming the original molecule in a more available secondary product. This study demonstrates for the first time that benthic macro-organisms can play an important role in the transformation of biodegradable contaminants, such as LAS.     

Investigations of the aufwuchs of freshwater sponges. I. A quantitative comparison between the surfaces of Spongilla Lacustris and three aquatic macrophytes

The aufwuchs of the epithelium of the freshwater sponge Spongilla lacustris was examined and compared qualitatively and quantitatively with that on three plant substrates common to its habitat: Nymphea odorata, Nuphar advena and Utricularia spp. Estimations of the percentage surface area with attached forms and the number of attached green algal filaments yielded significantly lower values for the sponge. Two theories explaining the mechanism effecting this reduced amount of aufwuchs are discussed.