Collagenic architecture and morphotraits in a marine basal metazoan as a model for bioinspired applied research.

In some Porifera (Demospongiae: Keratosa), prototypes of the connective system are almost exclusively based on collagenic networks. We studied the topographic distribution, spatial layout, microtraits, and/or morphogenesis of these collagenic structures in Ircinia retidermata (Dictyoceratida: Irciniidae). Analyses were carried out on a clonal strain from sustainable experimental mariculture by using light and scanning electron microscopy. Histology revealed new insights on the widely diversified and complex hierarchical assemblage of collagenic structures. Key evolutionary novelties in the organization of sponge connective system were found out. The aquiferous canals are shaped as corrugate-like pipelines conferring plasticity to the water circulation system. Compact clusters of elongated cells are putatively involved in a nutrient transferring system. Knob-ended filaments are characterized by a banding pattern and micro-components. Ectosome and outer endosome districts are the active fibrogenetic areas, where exogenous material constitutes an axial condensation nucleus for the ensuing morphogenesis. The new data can be useful to understand not only the evolutionary novelties occurring in the target taxon but also the morpho-functional significance of its adaptive collagenic anatomical traits. In addition, data may give insights on both marine collagen sustainable applied researches along with evolutionary and phylogenetic analyses, thus highlighting sponges as a key renewable source for inspired biomaterials. Therefore, we also promote bioresources sustainable exploitation with the aim to provide new donors of marine collagen, thereby supporting conservation of wild populations/species.

Biodiversity of Nearctic inland water: discovery of the genus Heterorotula (Porifera, Spongillida, Spongillidae) in the Appalachian Mountains, with biogeographical implications and description of new species.

This paper reports the discovery of a small population of sponges in the Pigeon River of eastern Tennessee, USA, which were morphologically distinct from Spongillida of North America. A morphological comparative analysis resulted in the first Nearctic record of the genus Heterorotula with the description of a new species Heterorotulalucasi sp. nov. diverging from all other known species by its unique combinations of diagnostic morphotraits of spicules and gemmules. The new record enlarges the geographic range of the genus which has been known until now only from Australia, New Zealand, New Caledonia, Japan (as an alien species), and from subequatorial Brazil (as subfossil remains). The discovery of a biogeographic enclave of Heterorotula in the southeastern United States contributes to the understanding of Porifera inland water biodiversity, biogeographic patterns, and adaptive morphotraits in the Nearctic and globally. Data confirm that the Appalachian region (Ordovician-Permian origin) of Tennessee and, in general, of North America have high levels of diversity and endemicity.

Relationship between bacterial phylotype and specialized metabolite production in the culturable microbiome of two freshwater sponges.

Microbial drug discovery programs rely heavily on accessing bacterial diversity from the environment to acquire new specialized metabolite (SM) lead compounds for the therapeutic pipeline. Therefore, knowledge of how commonly culturable bacterial taxa are distributed in nature, in addition to the degree of variation of SM production within those taxa, is critical to informing these front-end discovery efforts and making the overall sample collection and bacterial library creation process more efficient. In the current study, we employed MALDI-TOF mass spectrometry and the bioinformatics pipeline IDBac to analyze diversity within phylotype groupings and SM profiles of hundreds of bacterial isolates from two Eunapius fragilis freshwater sponges, collected 1.5 km apart. We demonstrated that within two sponge samples of the same species, the culturable bacterial populations contained significant overlap in approximate genus-level phylotypes but mostly nonoverlapping populations of isolates when grouped lower than the level of genus. Further, correlations between bacterial phylotype and SM production varied at the species level and below, suggesting SM distribution within bacterial taxa must be analyzed on a case-by-case basis. Our results suggest that two E. fragilis freshwater sponges collected in similar environments can exhibit large culturable diversity on a species-level scale, thus researchers should scrutinize the isolates with analyses that take both phylogeny and SM production into account to optimize the chemical space entering into a downstream bacterial library.

The genus Corvospongilla Annandale, 1911 (Porifera: Demospongiae: Spongillida) from Madagascar freshwater with description of a new species: biogeographic and evolutionary aspects.

We report here the first record of five genera of Spongillida (Porifera, Demospongiae) in both lentic and lotic water of Madagascar and the discovery of a new species of the genus Corvospongilla from the Betsiboka River hydrographic basin. Corvospongilla lemuriensis sp. n. diverges from all the other known species of the genus in its unique combination of diagnostic morphotraits (a) sessile gemmules with chambered pneumatic layer within tri-layered theca and enveloped by spicular cage of dominant acanthostrongyles, (b) dominant acanthostrongyles and less frequent acanthoxeas-strongyloxeas as megascleres and gemmuloscleres. The new species is characterised by the exclusive presence of a well-developed pneumatic layer in sessile gemmules not described for the genus until now. The new record confirms a Gondwanian track for the genus Corvospongilla and increases the knowledge on Spongillida historical biogeography. In synthesis the present discovery of Corvospongilla in the Malagasy biogeographic province (a) contributes to the assessment of Afrotropical biodiversity, (b) enlarges the geographic range of the genus reducing its disjunct distribution, and (c) focus on the evolutionary history of adaptive morphofunctional traits of resting bodies and life cycle timing in ephemeral water of the south-eastern Austral hemisphere.

Minimalist barcodes for sponges: a case study classifying African freshwater Spongillida.

African sponges, particularly freshwater sponges, are understudied relative to demosponges in most other geographical regions. Freshwater sponges (Spongillida) likely share a common ancestor; however, their evolutionary history, particularly during their radiation into endemic and allegedly cosmopolitan groups, is unclear. Freshwater sponges of at least 58 species of 17 genera and four families are described from Central and Eastern Africa, but the diversity is underestimated due to limited distinguishable morphological features. The discovery of additional cryptic species is very likely with the use of molecular techniques such as DNA barcoding. The Royal Museum of Central Africa (MRAC, Tervuren, Belgium) hosts one of the largest collections of (Central) African freshwater sponge type material. Type specimens in theory constitute ideal targets for molecular taxonomy; however, the success is frequently hampered by DNA degradation and deamination, which are a consequence of suboptimal preservation techniques. Therefore, we genotyped African demosponge holotype material of the MRAC with specific short primers suitable for degenerated tissue and compare the results with the current, morphology-based classification. Our results demonstrate the utility of minimalistic barcodes for identification of sponges, potentially enabling efficient identification of individuals in taxonomic or metabarcoding studies, and highlight inconsistencies in the current freshwater sponge classification.

Long-term experimental in situ farming of Crambe crambe (Demospongiae: Poecilosclerida).

BACKGROUND: The marine sponge Crambe crambe was chosen as an experimental model of sustainable shallow-water mariculture in the Sardinian Sea (Western Mediterranean) to provide biomass with high potential in applied research. METHODS: Explants were cultured in four long-term experiments (19 and 31 months at ca. 2.5 m depth), to determine the suitability of new culture techniques by testing substrata and seeding time (season), and monitoring survival and growth. Explants were excised and grown in an experimental plant close to the wild donor sponge population. Percentage growth rate (GR%) was measured in terms of surface cover area, and explant survival was monitored in situ by means of a digital photo camera. RESULTS: Explant survival was high throughout the trial, ranging from 78.57% to 92.85% on travertine tiles and from 50% to 71.42% on oyster shells. A few instances of sponge regression were observed. Explant cover area correlated positively with season on two substrata, i.e., tiles and shells. The surface cover area and GR% of explants were measured in the starting phase and monitored up to the end of the trial. High GR% values were observed both on tiles (>21%) and on oyster shells (>15%). DISCUSSION: The data on the behaviour and life-style of cultured fragments, together with an increase >2,400% in cover area, demonstrate that in situ aquaculture is a viable and sustainable method for the shallow-water biomass supply of Crambe crambe.

How Environmental Factors Affect the Production of Guanidine Alkaloids by the Mediterranean Sponge Crambe crambe.

Most marine sponges are known to produce a large array of low molecular-weight metabolites which have applications in the pharmaceutical industry. The production of so-called specialized metabolites may be closely related to environmental factors. In this context, assessing the contribution of factors like temperature, nutrients or light to the metabolomes of sponges provides relevant insights into their chemical ecology as well as the supply issue of natural sponge products. The sponge Crambe crambe was chosen as a model due to its high content of specialized metabolites belonging to polycyclic guanidine alkaloids (PGA). First results were obtained with field data of both wild and farmed specimens collected in two seasons and geographic areas of the North-Western Mediterranean. Then, further insights into factors responsible for changes in the metabolism were gained with sponges cultivated under controlled conditions in an aquarium. Comparative metabolomics showed a clear influence of the seasons and to a lesser extent of the geography while no effect of depth or farming was observed. Interestingly, sponge farming did not limit the production of PGA, while ex situ experiments did not show significant effects of several abiotic factors on the specialized metabolome at a one-month time scale. Some hypotheses were finally proposed to explain the very limited variations of PGA in C. crambe placed under different environmental conditions.

Natural collagenic skeleton of marine sponges in pharmaceutics: Innovative biomaterial for topical drug delivery.

The growing interest in the use of recyclable and biodegradable natural materials has become a relevant topic in pharmaceutics. In this work, we suggest the use and valorization of natural horny skeleton of marine sponges (Porifera, Dictyoceratida) as bio-based dressing for topical drug delivery. Biomaterial characterization focusing on morpho-functional traits, swelling behavior, fluid uptake performances, glycosaminoglycans content and composition and microbiological quality assessment was carried out to investigate the collagenic skeleton properties. After grinding and sieving processes, l-cysteine hydrochloride-loaded formulations were designed in form of powder or polymeric film by testing various drug concentrations and different drying parameters. Drug content, SEM analyses and in vitro permeation studies were performed to test the suitability of skeleton-based formulations. To this respect, drying time and temperature are key parameters for skeleton-mediated drug crystallization. Consequently, this behavior seems to influence drug loading and permeation profiles of formulations. The high percentages of drug are found after absorption into sponge powder and in vitro permeation studies demonstrate that cysteine is released more slowly than the pure drug within 1h. Such a system is attractive because it combines the known healing properties of cysteine with the advantageous potentials of the collagen/proteoglycan network, which can act as biocompatible carrier able to absorb the excess of the wound exudate while releasing the drug. Furthermore, due to its glycosaminoglycans content, natural sponge skeletal scaffold might act as bioactive-biomimetic carrier regulating the wound healing processes.

A new species of Agelas from the Zanzibar Archipelago, western Indian Ocean (Porifera, Demospongiae).

A new sponge species (Demospongiae: Agelasida: Agelasidae) is described from the eastern coast of Unguja Island in the Zanzibar Archipelago. Agelas sansibarica sp. n. is compared to all other Agelas species described so far. The new species differs from its congeners mainly in its three categories of verticillate spicules (acanthostyles, acanthostrongyles, and acanthoxeas) and their sizes. Acanthostrongyles, well represented in the spicular complement, are an exclusive trait of the new species widening the morphological range of the genus. Summarizing on spicular complement and spicular morphotraits of 36 species belonging to the genus Agelas: i) 32 species show only acanthostyles from Indo-Pacific (n = 14), Atlantic (n = 17), and Mediterranean (n = 1); ii) three Indo-Pacific species show acanthostyles and acanthoxeas; iii) one species Agelas sansibarica sp. n. from the western Indian Ocean is characterised by the unique trait of three categories of verticillate spicules (acanthostyles, acanthostrongyles and acanthoxeas). A key for the Indo-Pacific species is supplied together with short descriptions, illustrations, and geographic range; literature on chemical bioprospecting of the genus Agelas is also provided.

Swimming speed alteration in the early developmental stages of Paracentrotus lividus sea urchin as ecotoxicological endpoint.

Behavioral endpoints have been used for decades to assess chemical impacts at concentrations unlikely to cause mortality. With recently developed techniques, it is possible to investigate the swimming behavior of several organisms under laboratory conditions. The aims of this study were: i) assessing for the first time the feasibility of swimming speed analysis of the early developmental stage sea urchin Paracentrotus lividus by an automatic recording system ii) investigating any Swimming Speed Alteration (SSA) on P. lividus early stages exposed to a chemical reference; iii) identifying the most suitable stage for SSA test. Results show that the swimming speed of all the developmental stages was easily recorded. The swimming speed was inhibited as a function of toxicant concentration. Pluteus were the most appropriate stage for evaluating SSA in P. lividus as ecotoxicological endpoint. Finally, swimming of sea urchin early stages represents a sensitive endpoint to be considered in ecotoxicological investigations.

Discovery of living Potamolepidae (Porifera: Spongillina) from Nearctic freshwater with description of a new genus.

We report here the first record of a living Potamolepidae (Cherokeesia n. gen.) from the Nearctic Region and from the northern hemisphere. The new species Cherokeesia armata from southern Appalachians diverges at generic and specific level from all the other known taxa of the family Potamolepidae in its unique combination of diagnostic traits: gemmular theca armed by gemmuloscleres ranging from small strongyle-like spicules to stout, large oxeas; absence of pneumatic layer; spiny oxeas as main skeleton megascleres; irregular, slender pauci- to uni-spicular skeletal network. The most similar species belong to the genera Potamophloios and Oncosclera. The circumtropical biogeographic pattern of extant Potamolepidae, previously considered of Gondwanian to Gondwanian-like origin, is enlarged to the Nearctic. The present Tennessee discovery confirms a wider range of the family. An updated species inventory of Nearctic Spongillina, a checklist of the family Potamolepidae at the global level together with a key to the genera of Potamolepidae are also provided.

Adaptive morpho-traits, taxonomy and biogeography of Metania Gray, 1867 (Porifera: Spongillina: Metaniidae) with the description of a new species from Madagascar.

A comparative analysis of gemmular architecture adaptive morpho-traits at family level is reported for Metaniidae together with the discovery and description of a new species from the River Mangoky (High Plateau), Madagascar. The new Malagasy species, ascribed to Metania for diagnostic traits of the skeleton and the gemmular architecture, differs from all the other known species of the genus in its unique combination of diagnostic traits. Metania madagascariensis sp. nov. is characterised by encrusting growth form, conulose surface, specialized ectosomal skeleton, alveolate-reticulate choanosomal skeleton, two types of megascleres as smooth oxeas (α) and acanthoxeas (ß) ornamented with spines and/or tubercles, one type of microsclere as acanthoxeas with dense spines/tubercles bearing rosettes of microspines at tips; gemmules with or without cage of megascleres and frequently surrounded by microscleres; gemmular theca trilayered with pneumatic layer of fibrous spongin, boletiform (trumpet-like) gemmuloscleres with proximal true rotule large, smooth and with irregular blunt margins, and distal knob-like pseudorotule notably smaller, with a few hooks at the margins. M. madagascariensis belongs to the Afrotropical species group of Metania. Identification keys and an annotated checklist at global level are also provided together with a species-level discussion of Metania focusing on morphology, taxonomy, nomenclature and biogeography.

Isolation of hydroxyoctaprenyl-1',4'-hydroquinone, a new octaprenylhydroquinone from the marine sponge Sarcotragus spinosulus and evaluation of its pharmacological activity on acetylcholine and glutamate release in the rat central nervous system.

Three polyprenyl-1',4'-hydroquinone derivatives, heptaprenyl-1',4'-hydroquinone (1), octaprenyl-1',4'-hydroquinone (2), and hydroxyoctaprenyl-1',4'- hydroquinone (3) were isolated from the marine sponge Sarcotragus spinosulus collected at Baia di Porto Conte, Alghero (Italy). Our findings indicate that the compounds isolated from S. spinosulus can significantly modulate the release of glutamate and acetylcholine in the rat hippocampus and cortex and might, therefore, represent the prototype of a new class of drugs regulating glutamatergic and cholinergic transmission in the mammalian central nervous system.

An overview of the Mediterranean cave-dwelling horny sponges (Porifera, Demospongiae).

The present synthesis focuses on the so called 'horny sponges' recorded from marine caves of the Mediterranean Sea. The main aim is to provide a list of all recorded species, diagnostic keys to their identification up to family and genus level, and exhaustive, formally uniform descriptions at the species level contributing to sharing of information on the faunistics and taxonomy of Mediterranean cave-dwelling species, including habitat preferences. The majority of species was recorded in 105 Mediterranean marine caves hosting four orders of horny sponges belonging to 9 families, 19 genera and 40 species. Species endemic to the Mediterranean Sea harboured in marine caves are 14 with an endemicity value of 35%. For each species morphological descriptions are supported by illustrations both original and from the literature, including the diagnostic traits of the skeleton by light and scanning electron microscopy giving further characterization at the specific level. A detailed map together with a list of all caves harbouring horny sponges is also provided with geographic coordinates.

Sustainable production of biologically active molecules of marine based origin.

The marine environment offers both economic and scientific potential which are relatively untapped from a biotechnological point of view. These environments whilst harsh are ironically fragile and dependent on a harmonious life form balance. Exploitation of natural resources by exhaustive wild harvesting has obvious negative environmental consequences. From a European industry perspective marine organisms are a largely underutilised resource. This is not due to lack of interest but due to a lack of choice the industry faces for cost competitive, sustainable and environmentally conscientious product alternatives. Knowledge of the biotechnological potential of marine organisms together with the development of sustainable systems for their cultivation, processing and utilisation are essential. In 2010, the European Commission recognised this need and funded a collaborative RTD/SME project under the Framework 7-Knowledge Based Bio-Economy (KBBE) Theme 2 Programme 'Sustainable culture of marine microorganisms, algae and/or invertebrates for high value added products'. The scope of that project entitled 'Sustainable Production of Biologically Active Molecules of Marine Based Origin' (BAMMBO) is outlined. Although the Union is a global leader in many technologies, it faces increasing competition from traditional rivals and emerging economies alike and must therefore improve its innovation performance. For this reason innovation is placed at the heart of a European Horizon 2020 Strategy wherein the challenge is to connect economic performance to eco performance. This article provides a synopsis of the research activities of the BAMMBO project as they fit within the wider scope of sustainable environmentally conscientious marine resource exploitation for high-value biomolecules.

Effects of Agelas oroides and Petrosia ficiformis crude extracts on human neuroblastoma cell survival.

Among marine sessile organisms, sponges (Porifera) are the major producers of bioactive secondary metabolites that defend them against predators and competitors and are used to interfere with the pathogenesis of many human diseases. Some of these biological active metabolites are able to influence cell survival and death, modifying the activity of several enzymes involved in these cellular processes. These natural compounds show a potential anticancer activity but the mechanism of this action is largely unknown. In this study, we investigated the effects of two Mediterranean sponges, Agelas oroides and Petrosia ficiformis on the viability of human neuroblastoma cells. Upon treatment with the methanolic extract of Petrosia ficiformis, a marked cytotoxic effect was observed at any concentration or time of exposure. In contrast, a time- and dose-dependent effect was monitored for Agelas oroides that induced the development of apoptotic features and ROS production in LAN5 cells. These events were suppressed by calpeptin or zVAD and by vitamin C suggesting that the cell death caused by Agelas oroides was calpain- and caspase-dependent and of oxidative nature. Comet assay showed that this methanolic extract was not able to produce a genotoxic effect. Future studies will be applied to investigate the effect of isolated bioactive compounds from crude extract of this sponge which are potentially useful for cancer therapeutics.

The marine sponge Chondrilla nucula Schmidt, 1862 as an elective candidate for bioremediation in integrated aquaculture.

The use of sponges for marine bioremediation in a farming scenario has been investigated focusing on Chondrilla nucula. We report experiments examining clearance and retention rates of the bacterium Escherichia coli. Despite low values expressed for clearance tests, C. nucula exhibited a marked ability to retain high quantities of bacteria. One square meter patch of this sponge can filter up to 14 l/h of sea water retaining up to 7 x 10(10) bacterial cells/h. This suggests that C. nucula is a suitable species for marine environmental bioremediation.