Mediterranean Bioconstructions Along the Italian Coast.

Marine bioconstructions are biodiversity-rich, three-dimensional biogenic structures, regulating key ecological functions of benthic ecosystems worldwide. Tropical coral reefs are outstanding for their beauty, diversity and complexity, but analogous types of bioconstructions are also present in temperate seas. The main bioconstructions in the Mediterranean Sea are represented by coralligenous formations, vermetid reefs, deep-sea cold-water corals, Lithophyllum byssoides trottoirs, coral banks formed by the shallow-water corals Cladocora caespitosa or Astroides calycularis, and sabellariid or serpulid worm reefs. Bioconstructions change the morphological and chemicophysical features of primary substrates and create new habitats for a large variety of organisms, playing pivotal roles in ecosystem functioning. In spite of their importance, Mediterranean bioconstructions have not received the same attention that tropical coral reefs have, and the knowledge of their biology, ecology and distribution is still fragmentary. All existing data about the spatial distribution of Italian bioconstructions have been collected, together with information about their growth patterns, dynamics and connectivity. The degradation of these habitats as a consequence of anthropogenic pressures (pollution, organic enrichment, fishery, coastal development, direct physical disturbance), climate change and the spread of invasive species was also investigated. The study of bioconstructions requires a holistic approach leading to a better understanding of their ecology and the application of more insightful management and conservation measures at basin scale, within ecologically coherent units based on connectivity: the cells of ecosystem functioning.

Life-cycle traits in the demosponge Hymeniacidon perlevis in a land-based fish farm.

Background: The demosponge Hymeniacidon perlevis is characterized by wide geographic distribution and great adaptability to numerous and highly variable climatic and hydrological conditions. Indeed, the species can colonize many different environments, including several unusual ones, such as concrete drainage conduits of a marine land-based fish farm plant. This research aimed to enhance existing knowledge on the reproductive cycle and growth performance of H. perlevis while also evaluating the impact of a controlled supply of trophic resources, wastewater flow and constant water temperature on these biological traits. Methods: Specimens included in this one-year study inhabited drainage conduits of a land-based fish farm. The approach included measurements of sponge biomass and occurrence and abundance of reproductive elements across different seasons and environmental parameters, such as fish biomass, trophic resources, and wastewater flow. Sponge growth and reproductive elements, including oocytes, spermatic cysts, and embryos, were measured monthly in sponges positioned in the drainage conduit, thus with different trophic resources but with constant water temperature. Finally, we used generalized additive models to describe variables that contribute the most to the growth of sponges. Results: Growth performance showed marked variations during the study period. The highest increase in sponge volume was observed from August/September to January/March. The volume of sponges was principally determined by the reduction of reared fish biomass and the increase of pellet amount and wastewater flow. Sponge specimens exhibited an active state during the entire study, as proven by the occurrence of recruits. However, sexual elements were only sporadically observed, thus not permitting the recognition of a true sexual cycle. Discussion: The results of the present study confirmed that H. perlevis exhibits high flexibility and adaptability to the differential, and somewhat extreme, environmental conditions. Indeed, this species can live, grow and reproduce in the drainage conduits of the fish farm, where the species face constant darkness, water temperature and continuous nutritional supply. In such conditions, H. perlevis display an active state during the entire year, while avoiding stages of decline and long dormancy usually observed in wild populations. It seems plausible that stable environmental conditions induce an almost continuous sexual phase, probably under the control of endogenous factors. No asexual elements were detected, although it was impossible to exclude the contribution of asexual reproduction in the origin of the newly settled sponges, which were repeatedly detected throughout the study. The growth performance seemed linked to the fish farm conditions, thus providing useful indications on the best maintenance conditions for H. perlevis in land-based integrated multitrophic systems, where the species could be used for wastewater treatment.

Alien and native species in Italian marine and transitional waters.

Background: Biological invasions are one of the major threats to the ecosystem structure and functioning. After the initial introduction, frequently mediated by human activities, alien species can overcome different biogeographical and ecological barriers and determine severe impacts on native biodiversity and socio-economic activities. The Italian peninsula is located at the intersection of large trade routes within the Mediterranean Sea. Such position, along with the intense commercial activity and the high population density of the Italian coast, are considered important drivers of alien species in Italian marine and transitional ecosystems. The Italian peninsula, however, is also one of the regions with the highest native species richness within the Mediterranean Sea and, therefore, it is crucial to account for both alien and native species diversity when estimating the impact of biological invasion. Yet, such comprehensive information is frequently scattered across several biodiversity information systems and databases. New information: Here, two datasets with alien and native species records in Italian marine and transitional waters are described. These datasets, created for the LifeWatch Italy case study on alien species, are the result of a large-scale collaboration involving experts working across the whole range of taxonomic diversity. The marine dataset includes a total of 12,219 records belonging to 3,772 species gathered from 91 investigated sites and seven EUNIS habitats. The dataset on transitional waters biodiversity includes 3,838 records belonging to 2,019 species found in 23 locations and four EUNIS habitats. Alien species were recorded in both marine and transitional waters, accounting respectively for 140 and 171 biological records belonging to 59 and 97 species. These occurrence data can be used for further research studies or management purposes, including the evaluation of the invasion risk and the formulation of alien species control and management plans. Furthermore, these compiled datasets can be used as input data for the Biotope vulnerability case study of LifeWatch ERIC, which offers a number of ICT services for the calculation of the incidence and of the impact of alien species on European biotopes.

Epidemic mortality of the sponge Ircinia variabilis (Schmidt, 1862) associated to proliferation of a Vibrio bacterium.

In recent years, several episodes of mass mortality of sessile epibenthic invertebrates, including sponges, have been recorded worldwide. In the present study, we report a disease event on Ircinia variabilis recorded in September 2009 along the southern Adriatic and Ionian seas (Apulian coast), with the aim to quantify the mortality incidence on the sponge population, to investigate the effect of the disease on the sponge tissues and to assess whether the disease is associated with vibrios proliferation. The injured sponges showed wide necrotic areas on the surface or disruption of the body in several portions. Necrotic areas were whitish and often were covered with a thin mucous coat formed by bacteria. In the most affected specimens, sponge organisation resulted partial or complete loss, with the final exposure of the dense skeletal network of spongine fibres to the environment. The results of microbiological cultural analysis using in parallel Marine Agar 2216 and thiosulphate/citrate/bile salts/sucrose agar demonstrated that, in affected specimens, vibrios represented 15.8 % of the total I. variabilis surface culturable bacteria. Moreover, all the isolated vibrios, grown from the wide whitish areas that characterize the surface of the diseased sponges, were identified, and their assignment to the Vibrio rotiferianus was consistent with phylogenetic analysis and data of morphological, cultural and biochemical tests. Studies on V. rotiferianus have shown that its pathogenicity, with respect to various aquatic organisms, is higher than that of Vibrio harveyi. The factors triggering the disease outbreak in Ircinia variabilis populations remain unclear. At present, we can hypothesize the involvement in the disease of a synergetic mechanism that, under stressful physiological conditions (high temperature, elevated nutrients and reduced water flow), induces sponge pathogens, in our case V. rotiferanius, to become virulent, making sponges unable to control their proliferation. Additional studies are needed to understand the etiological processes as well as the factors involved in sponges recovering from this epidemic event allowing them to face mass mortality. A drastic reduction of sponge-specific representatives could have marked a negative impact on the environmental health on account of their role in the sea remediation processes as filter-feeding organisms.

Large-Scale Distribution of the European Seahorses (Hippocampus Rafinesque, 1810): A Systematic Review.

Human pressures on marine ecosystems have caused extensive degradation of marine habitats and several local extinctions. Overexploitation and destructive fishing practices are responsible for biodiversity loss in many coastal ecosystems. The definition of conservation programs in marine fish requires comprehensive knowledge on large-scale geographical distribution, while considering distribution/abundance patterns in relation to key environmental variables. Due to their life-cycle traits, the two European seahorses (Hippocampus guttulatus and H. hippocampus), as with other congeneric species, are particularly sensitive to the effects of anthropogenic activities and habitat changes. However, information on the ecological distribution of these two species is scattered, patchy, and mainly focused on small-scale studies. In this paper, we followed an international standard protocol for systematic reviews (the PRISMA protocol) to provide a detailed assessment of the two species' geographical distribution in relation to the environmental characteristics. According to the 134 analyzed studies, Hippocampus guttulatus is more common in confined areas, while H. hippocampus is found in marine shelf waters. With several interspecific differences, seagrasses were the most used holdfasts of both species. The EUNIS codes (European nature information system) referring to a specific and unique habitat were discussed as a potential tool for defining the ecological distribution of the two species. The obtained results and their future implementation could help plan conservation actions.

Effects and Impacts of Different Oxidative Digestion Treatments on Virgin and Aged Microplastic Particles.

Although several sample preparation methods for analyzing microplastics (MPs) in environmental matrices have been implemented in recent years, important uncertainties and criticalities in the approaches adopted still persist. Preliminary purification of samples, based on oxidative digestion, is an important phase to isolate microplastics from the environmental matrix; it should guarantee both efficacy and minimal damage to the particles. In this context, our study aims to evaluate Fenton's reaction digestion pre-treatment used to isolate and extract microplastics from environmental matrices. We evaluated the particle recovery efficiency and the impact of the oxidation method on the integrity of the MPs subjected to digestion considering different particles' polymeric composition, size, and morphology. For this purpose, two laboratory experiments were set up: the first one to evaluate the efficacy of various digestion protocols in the MPs extraction from a complex matrix, and the second one to assess the possible harm of different treatments, differing in temperatures and volume reagents used, on virgin and aged MPs. Morphological, physicochemical, and dimensional changes were verified by Scanning Electron Microscope (SEM) and Fourier Transformed Infrared (FTIR) spectroscopy. The findings of the first experiment showed the greatest difference in recovery rates especially for polyvinyl chloride and polyethylene terephthalate particles, indicating the role of temperature and the kind of polymer as the major factors influencing MPs extraction. In the second experiment, the SEM analysis revealed morphological and particle size alterations of various entities, in particular for the particles treated at 75 °C and with major evident alterations of aged MPs to virgin ones. In conclusion, this study highlights how several factors, including temperature and polymer, influence the integrity of the particles altering the quality of the final data.

Sexual and asexual reproduction in a Mediterranean Tethya (Porifera, Demospongiae) species.

BACKGROUND: The reproductive cycle of the recently described sponge Tethya meloni was investigated for a period of 15 months (September 2018 - November 2019) in the Mar Piccolo of Taranto (Southern Italy) and was compared with data previously collected for the other two sympatric species of the same genus known for Mediterranean Sea, T. citrina and T. aurantium. RESULTS: T. meloni is a gonochoric species with a sex ratio strongly shifted towards females. Asexual budding was a seasonal process, limited to few specimens. In a specimen collected in September 2018 both oocytes and buds occurred, suggesting that in T. meloni the sexual and asexual phases may coexist both at the population and individual levels. CONCLUSIONS: The data obtained from this research compared with the available literature confirm the high temporal variability of the reproductive cycles in the Mediterranean species of Tethya, but with common general characteristics. In sexual reproduction, the oocyte production period lasts several months, with a peak between summer and autumn while spermatogenesis, shorter but with greater reproductive effort, follows the onset of oogenesis. The asexual reproduction phase of T. meloni, on the other hand, occurs in a short period and seems to have less importance in the overall reproductive process.

Evaluating the Efficiency of DNA Metabarcoding to Analyze the Diet of Hippocampus guttulatus (Teleostea: Syngnathidae).

Seahorses are considered a flagship species for conservation efforts and due to their conservation status, improving knowledge on their dietary composition while applying a non-invasive approach, could be useful. Using Hippocampus guttulatus as a case study, the present study represents pioneering research into investigating the diet of seahorses by NGS-based DNA metabarcoding of fecal samples. The study developed and tested the protocol for fecal DNA metabarcoding during the feeding trials where captive seahorses were fed on a diet of known composition; the process was subsequently applied on fecal samples collected from wild individuals. The analysis of samples collected during the feeding trials indicated the reliability of the applied molecular approach by allowing the characterization of the effectively ingested prey. In the field study, among detected prey species, results revealed that the majority of the seahorse samples contained taxa such as Amphipoda, Decapoda, Isopoda, and Calanoida, while less common prey taxa were Gastropoda and Polyplacophora. As only a small amount of starting fecal material is needed and the sampling procedure is neither invasive nor lethal. The present study indicates DNA metabarcoding as useful for investigating seahorse diet and could help define management and conservation actions.

Massive bioconstructions built by Neopycnodonte cochlear (Mollusca, Bivalvia) in a mesophotic environment in the central Mediterranean Sea.

The present paper provides a multidisciplinary fine-scale description of a Mediterranean mesophotic new habitat dominated by the bivalve Neopycnodonte cochlear (Poli, 1795), building large and thick pinnacles on vertical cliffs at two study areas along the southern Italian coast. The pinnacles, constituted by a multilayered aggregation of living and dead specimens of N. cochlear, were interconnected with each other to form a framework of high structural complexity, never observed before for this species. The bioconstruction, considerably extended, resulted very complex and diversified in the associated community of structuring organisms. This latter included 165 taxa attributable to different ecological groups occurring in different microhabitats of the bioconstruction. Among the secondary structuring taxa there were scleractinians, serpulids and bryozoans, all contributing to the deposition of calcium carbonate, and poriferans, helping to bind shells together or eroding carbonate by boring species. In comparison with coralligenous sensu stricto and the recently described Mediterranean mesophotic coral reef, the Neopycnodonte bioconstruction showed peculiar features, since it lacked the major contribution of encrusting coralline algae and scleractinians as reef builders, respectively.

Spinal glioneuronal tumor with neuropil-like islands and meningeal dissemination: histopathological and radiological study of a pediatric case.

Cerebral and spinal location of glioneuronal tumors have been recently described as a novel type of primary CNS neoplasia. A distinctive rare form of glioneuronal tumors with neuropil-like islands (GTNI) have been reported to occur in the adult cerebrum, whereas spinal GTNI localization is extremely rare. In the present report we describe a case of a 15-month-old child with a spinal GTNI of the cervical region and meningeal dissemination. Histologically the tumor was composed of round, small neurocytic-like cells arranged around eosinophilic neuropil cores and embedded in a diffuse fibrillar glial component forming prominent "rosetted" neuropil islands displaying strong immunoreactivity for neuronal markers. Cerebral GTNI shows abundant glial components not rarely exhibiting anaplastic features that justify their inclusion within the group of diffuse astrocytomas. In contrast, including our case, spinal GTNI do not show histological evidence of anaplastic features and exhibits a significant neuronal component that may imply considering these lesions in a separate group. Nevertheless, due to their exceptional rarity, the natural history of these lesions is not yet fully understood, but spinal GTNI seems to have an unfavorable clinical course despite their benign histopathological features, which must be taken into account for appropriate treatment and follow-up of the patient.

Two new species of deep-sea sponges (Porifera, Demospongiae) from submarine canyons of the Sardinian continental margin (western Mediterranean Sea).

Here we describe two new species of deep-sea sponges collected during the exploration of Cold-Water Coral (CWC) banks discovered in the Nora and Coda Cavallo canyons (north-eastern and south coast of Sardinia respectively). Poecillastra tavianii n. sp. differs from the other congeneric species mainly for the dicho- and mesodichotriaenes, never observed in the genus, and the abundance and variety of spirasters. Hymedesmia (Hymedesmia) quadridentata n. sp. is characterized by the presence of two types of chelae. In particular, the unguiferate chelae (round shaft and four teeth at both extremities) represent a peculiar character of the species. Our contribution increases the number of sponge fauna associated to the best known Central Mediterranean CWC habitats to 98 improving the still scant knowledge on the biodiversity of the Mediterranean CWC habitats.

Data on the diet composition of Hippocampus guttulatus cuvier, 1829: Different prey preferences among habitats.

The data presented here support research article entitled 'Trophic flexibility and prey selection of the wild long-snouted seahorse Hippocampus guttulatus Cuvier, 1829 in three coastal habitats'' Ape et al., 2019. Determinations of the dietary composition, differences in prey selection and potential prey abundance and availability among three habitats at Taranto Mar Piccolo were based on the analysis of gut contents of seahorses and sediment samples. Both highly (Corallina elongata and Cladophora prolifera) and low complex (sandy bottom) habitats were investigated. Prey items were divided into two size classes: <1 mm and >1mm. Data about the total abundance of each prey size class in gut contents and sediments in three different habitats and PERMANOVA comparisons are given.

A Mediterranean mesophotic coral reef built by non-symbiotic scleractinians.

This is the first description of a Mediterranean mesophotic coral reef. The bioconstruction extended for 2.5 km along the Italian Adriatic coast in the bathymetric range -30/-55 m. It appeared as a framework of coral blocks mostly built by two scleractinians, Phyllangia americana mouchezii (Lacaze-Duthiers, 1897) and Polycyathus muellerae (Abel, 1959), which were able to edify a secondary substrate with high structural complexity. Scleractinian corallites were cemented by calcified polychaete tubes and organized into an interlocking meshwork that provided the reef stiffness. Aggregates of several individuals of the bivalve Neopycnodonte cochlear (Poli, 1795) contributed to the compactness of the structure. The species composition of the benthic community showed a marked similarity with those described for Mediterranean coralligenous communities and it appeared to be dominated by invertebrates, while calcareous algae, which are usually considered the main coralligenous reef-builders, were poorly represented. Overall, the studied reef can be considered a unique environment, to be included in the wide and diversified category of Mediterranean bioconstructions. The main reef-building scleractinians lacked algal symbionts, suggesting that heterotrophy had a major role in the metabolic processes that supported the production of calcium carbonate. The large amount of available suspended organic matter in the area could be the main nutritional source for these species, as already suggested in the literature referred to Mediterranean cold-water corals.

Evaluation of microbiological accumulation capability of the commercial sponge Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae).

This study was carried out to evaluate the microbiological accumulation capability of the demosponge Spongia officinalis var. adriatica. Six microbiological parameters were researched in two sampling periods in the water and in reared sponge samples coming from sites with different degrees of microbial contamination: an off-shore fish farm displaced off the Apulian coast (Southern Adriatic Sea) and a no-impacted area displaced into the Marine Protected Area of Porto Cesareo (Apulian coast-Ionian Sea). We detected the density of culturable heterotrophic bacteria by spread plate on marine agar, total culturable bacteria at 37 degrees C on Plate Count Agar and vibrios on thiosulphate-citrate-bile-sucrose-salt (TCBS) agar. Total and fecal coliforms as well as fecal streptococci concentrations were detected by the MPN method. Bacterial densities were always higher in the sponge homogenates compared with the corresponding seawater in the sampling points and in both sampling periods. As regard vibrios, total culturable bacteria at 37 degrees C and fecal streptococci concentrations, the highest values were observed in the sponge samples coming from the off-shore fish farm during the summer period. The ability of Spongia officinalis var. adriatica to accumulate the microbial pollution indicators suggests that this species can be employed as a bioindicator for monitoring water quality.

Filtering activity of Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae) on bacterioplankton: implications for bioremediation of polluted seawater.

A study on the filtering activity has been carried out on reared specimens of the demosponge Spongia officinalis var. adriatica coming from an off-shore farm displaced off the Apulian coast (Ionian Sea). The experience was carried out under laboratory conditions, by using natural seawater collected from the sponge environment. The study demonstrates a high efficiency of the sponge in removing bacteria. Bacterial concentration significantly decreases in presence of the sponge, with a marked drop after 2 h from the start of the experience. The maximum clearance rate was 210 ml h(-1) g(-1) DW at 60 min. Retention efficiency reached the highest value of 61% at 120 min. The bacterial density removed by the S. officinalis filtering activity was 12.3 +/- 1.8 x 10(4) cells ml(-1) corresponding to a biomass of about 11.7 +/- 1.4 microg Cl(-1). The sponge fed preferentially large- and medium-size bacteria, whereas the small ones are fed after the removal of the largest size categories. The results obtained suggest that S. offcinalis is a suitable species for marine environmental bioremediation.

The co-occurrence of the demosponge Hymeniacidon perlevis and the edible mussel Mytilus galloprovincialis as a new tool for bacterial load mitigation in aquaculture.

Pollutants in marine coastal areas are mainly a consequence of anthropogenic inputs, and microorganisms often play a major role in determining the extent of this pollution. Thus, practical and eco-friendly techniques are urgently required in order to control or minimise the pathogenic bacterial problem. The bacterial accumulation of Mytilus galloprovincialis (Lamarck 1919) in the presence or absence of another filter feeder, the demosponge Hymeniacidon perlevis (Montagu 1818) on sewage flowing into the Northern Ionian Sea has been estimated in a laboratory study. On account of the interesting results obtained, we also evaluated the bioremediation capability of the sponges when reared in co-culture with mussels. Specimens of M. galloprovincialis and H. perlevis were collected from the Mar Grande and from the Second Inlet of the Mar Piccolo of Taranto (Northern Ionian Sea, Italy), respectively. In the laboratory, we detected the bacterial abundances in the sewage, in sponge homogenates (both sponges alone and sponges that have been added to sewage with mussels) and in mussel homogenates (both mussels alone and mussels that have been added to sewage with sponges). In the field, we estimated the bacterial concentration in both the seawater within the mussels culture and the seawater collected where mussels were reared in co-culture with sponges. The bacteriological analyses were performed analysing the following parameters: the density of culturable heterotrophic bacteria by spread plate on marine agar, total culturable bacteria at 37 °C on plate count agar and vibrios on thiosulphate-citrate-bile-sucrose-salt (TCBS) agar. Total coliforms, Escherichia coli and intestinal streptococci concentrations were detected by the MPN method. The study demonstrates a higher efficiency of the sponges in removing all the considered bacterial groups compared to the mussels. Due to the conspicuous bacterial accumulation by the sponge, we can conclude that the co-occurrence of the filter-feeder H. perlevis with M. galloprovincialis is a powerful tool in reducing the bacterial load in shellfish culture areas thus playing a role in mitigating the health hazard related to the consumption of edible mussels.

Analytical investigations on the lindane bioremediation capability of the demosponge Hymeniacidon perlevis.

Lindane is an organochlorine pesticide that has been widely used to treat agricultural pests. It is of particular concern because of its toxicity, persistence and tendency to bioaccumulate in terrestrial and aquatic ecosystems. In this context, we investigated the ability of the demosponge Hymeniacidon perlevis to bioremediate lindane polluted seawater during in vitro experimentation. Lindane was extracted by solid-phase micro-extraction (SPME) and determined by gas chromatography-mass spectrometry (GC-MS). Furthermore, we assessed the role exerted in lindane degradation by bacteria isolated from the sponge. Sponges showed low mortality in experimental conditions (lindane concentration 1 µg/L) and were able to remove about 50% of the lindane content from seawater in 48 h. Bacteria isolated from sponges showed a remarkable remediating capacity (up to 97% of lindane removed after 8-days). A lindane metabolite was identified, 1,3,4,5,6-pentachloro-cyclohexene. The results obtained are a prelude to the development of future strategies for the in situ bioremediation of this pollutant.

Inhibitory Effect of Aqueous Extracts from Marine Sponges on the Activity and Expression of Gelatinases A (MMP-2) and B (MMP-9) in Rat Astrocyte Cultures.

The aim of this study was to evaluate whether water soluble compounds present in aqueous extracts from seven Mediterranean demosponges exert biological activity towards matrix metalloproteinases (MMPs), which represent important pathogenic factors of human diseases. Aqueous extracts were tested on LPS-activated cultured rat astrocytes, and levels and expression of MMP-2 and MMP-9 were assessed by zymography and RT-PCR, respectively. Our results demonstrated that the studied extracts contain water soluble compounds able to inhibit MMP-2 and MMP-9 activity and expression. We also compared the anti-MMP activities present in aqueous extracts from wild and reared specimens of Tethya aurantium and T. citrina. The results obtained revealed that the reared sponges maintain the production of bioactive compounds with inhibitory effect on MMP-2 and MMP-9 for all the duration of the rearing period. Taken together, our results indicate that the aqueous extracts from the selected Mediterranean demosponges possess a variety of water-soluble bioactive compounds, which are able to inhibit MMPs at different levels. The presence of biological activity in aqueous extracts from reared specimens of T. aurantium and T. citrina strongly encourage sponge aquaculture as a valid option to supply sponge biomass for drug development on a large scale.

Comparative analysis of protein profiles of aqueous extracts from marine sponges and assessment of cytotoxicity on different mammalian cell types.

Marine natural products extracted from sponges represent a new source for drug discovery. Here we describe a simple method for preparing aqueous extracts from 7 Mediterranean demosponges, which allowed the extraction of water-soluble compounds, such as proteins by homogenization of sponge tissue in phosphate buffered saline (PBS). The comparative analysis by SDS-PAGE showed differences in number of bands, bandwidth and intensity among the sponges analyzed. The PAS/silver staining revealed a substantial and different glycoprotein assortment among the demosponges studied. To further study the biological activities present in the sponge extracts, we determined the non-cytotoxic doses on four different mammalian cell types demonstrating that the optimal non-cytotoxic doses were cell type- and extract-dependent. In conclusion, the extraction method described in this paper represents a fast and efficient procedure for the extraction of water-soluble proteins from marine sponges. Furthermore, the cell viability data suggest the feasibility of this method for the direct in vitro cell-based assays.

Life-cycle traits of Paraleucilla magna, a calcareous sponge invasive in a coastal Mediterranean Basin.

The calcareous sponge Paraleucilla magna, originally observed along the Brazilian coast (Atlantic Ocean), is the only allochthonous invasive species of Porifera reported in the Mediterranean Sea. A 1-year investigation of the population dynamics and life-cycle of this exotic species in the Mar Piccolo di Taranto (southern Italy, central Mediterranean Sea) has provided a good opportunity to test how environmental variations can influence its life-cycle and to ascertain what strategy can be adopted to successfully colonize a new environment. In the Mar Piccolo di Taranto, P. magna exhibits marked temporal changes in biomass. The studied specimens reproduce almost all year round, showing a seasonal pattern that peaks during warm months. This prolonged sexual activity allows P. magna to continuously produce young specimens, with repeated recruitment events taking place throughout the year, thus offsetting the seasonal mortality of adult specimens. This r-strategy enables the non-indigenous sponge to achieve a high degree of maintenance over relatively long periods (ten years at least).