A bioreaction–diffusion model for growth of marine sponge explants in bioreactors

Marine sponges are sources of high-value bioactives. Engineering aspects of in vitro culture of sponges from cuttings (explants) are poorly understood. This work develops a diffusion-controlled growth model for sponge explants. The model assumes that the explant growth is controlled by diffusive transport of at least some nutrients from the surrounding medium into the explant that generally has a poorly developed aquiferous system for internal irrigation during early stages of growth. Growth is assumed to obey Monod-type kinetics. The model is shown to satisfactorily explain the measured growth behavior of the marine sponge Crambe crambe in two different growth media. In addition, the model is generally consistent with published data for growth of explants of the sponges Disidea avara and Hemimycale columella. The model predicted that nutrient concentration profiles for nutrients, such as dissolved oxygen within the explant, are consistent with data published by independent researchers. In view of the proposed model’s ability to explain available data for growth of several species of sponge explants, diffusive transport does play a controlling role in explant growth at least until a fully developed aquiferous system has become established. According to the model and experimental observations, the instantaneous growth rate depends on the size of the explant and all those factors that influence the diffusion of critical nutrients within the explant. Growth follows a hyperbolic profile that is consistent with the Monod kinetics.     

Hypothesized Kinetic Models for Describing the Growth of Globular and Encrusting Demosponges

The marine sponges Dysidea avara and Chondrosia reniformis (globular forms) were cultured in the laboratory on a diet of viable Phaeodactylum tricornutum cells and dissolved nutrients (algae and fish powders). Our growth data were combined with literature data for Pseudosuberites andrewsi (a globular sponge) and for the encrusting sponges Oscarella lobularis, Hemimycale columella, and Crambe crambe. The suitability of three growth models—linear, exponential, and radial accretive—for describing the growth of globular and encrusting sponges was assessed. Radial accretive growth was determined to be the best model to describe growth of both encrusting and globular sponges. Average growth rates of 0.051 ± 0.016 and 0.019 ± 0.003 mm/day (calculated as the increase of the radius of the sponge per day) were obtained experimentally for D. avara and C. reniformis, respectively.     

Dispersal strategies in sponge larvae: integrating the life history of larvae and the hydrologic component

While known to be uniformly non-feeding, short-lived, and potentially short dispersing, sponge larvae display different behaviours (swimming ability and taxis). Our aim was to show whether sponge larvae with different behaviours exhibit different dispersal strategies under variable intensity of water movements. We first assessed the distribution of larvae of six taxa: Dictyoceratida spp., Dysidea avara, Crambe crambe, Phorbas tenacior, Scopalina lophyropoda, and Cliona viridis, collected through plankton sampling, and the abundance of the corresponding adult sponges across three hard bottom communities and a sandy bottom from a north-west Mediterranean rocky shore. We then tested adult–larvae couplings (abundance of larvae vs abundance of adults) under increasing levels of water movements (surge) to assess the importance of this environmental factor in driving differences in dispersal strategies. Adults of Dictyoceratida spp., D. avara, and P. tenacior were most abundant in semi-dark caves (SDC), C. crambe and C. viridis in communities of sciaphilic algae (SA), whereas the distribution of S. lophyropoda was extremely patchy, being present almost only in the SA community of one of the five stations studied. Larvae of Dictyoceratida spp. and P. tenacior were more abundant in the SDC, whereas D. avara and C. crambe were homogeneously distributed across the communities. The larvae of C. viridis were more abundant in the SA communities and the S. lophyropoda larvae were mostly present in one station and one community (SA). Increased water movement did not modify the adult–larvae coupling for Dictyoceratida spp., D. avara, and C. crambe, whereas it broke up the positive association for P. tenacior and to some extent S. lophyropoda. For C. viridis, possible variability in adult–larvae coupling was not tested because the larvae were collected on only one day under calm sea conditions. We confirm that efficient-swimming larvae with some cue response can actively counteract hydrodynamic forces and highlight the importance of both larval behaviour and environmental conditions in determining small-scale patterns of dispersal.     

Cultivation of Sponge Larvae: Settlement,Survival, and Growth of Juveniles

The aim of this study was to culture sponge juveniles from larvae. Starting from larvae we expected to enhance the survival and growth, and to decrease the variation in these parameters during the sponge cultures. First, settlement success, morphological changes during metamorphosis, and survival of Dysidea avara, Ircinia oros, Hippospongia communis, under the same culture conditions, were compared. In a second step, we tested the effects of flow and food on survival and growth of juveniles from Dysidea avara and Crambe crambe. Finally, in a third experiment, we monitored survival and growth of juveniles of D. avara and C. crambe transplanted to the sea to compare laboratory and field results. The results altogether indicated that sponge culture from larvae is a promising method for sponge supply and that laboratory culture under controlled conditions is preferred over sea cultures in order to prevent biomass losses during these early life stages.     

The choanoderm of Sycettusa hastifera (Calcarea,Porifera) is able to generate new individuals

One of the main characteristics of sponges is their capacity for cell dedifferentiation. This capability can allow an impressive amount of asexual reproduction in these animals, because they are able to develop new individuals from just a few somatic cells. Studies of dedifferentiation, however, have focused mainly on sponges of the class Demospongiae. Therefore, we investigated here whether individuals of three different species of Calcarea are able to reconstitute new individuals following artificial fragmentation. We observed that fragmentation releases clumps of choanoderm able to initiate somatic embryogenesis. In Borojevia brasiliensis (asconoid aquiferous system, subclass Calcinea) and Paraleucilla magna (leuconoid aquiferous system, subclass Calcaronea), these clumps started to develop, but they did not pass through the first developmental phases. In Sycettusa hastifera (syconoid aquiferous system, subclass Calcaronea), the choanoderm was reorganized into primmorphs that fused to each other and formed an exopinacoderm. The first primmorphs’ spicules were triactines. Despite a large mortality rate, the primmorphs developed into olynthus stages. The somatic embryogenesis and the metamorphosis of the olynthus were similar to those observed during the sexual development of this and other calcareous sponge species. Our results show that in S. hastifera, and perhaps in other syconoid calcareous sponges, somatic embryogenesis occurs mainly from choanocytes, at least in vitro. However, primmorph development does not follow the same pattern observed in post‐metamorphic sexual development, as in that case diactines are always the first spicules to be synthesized in calcaronean species.     

16S rDNA clone library-based bacterial phylogenetic diversity associated with three South China Sea sponges

Culture-independent molecular techniques, 16S rDNA clone library alongside RFLP and phylogenetic analysis, were applied to investigate the bacterial diversity associated with three South China Sea sponges, Stelletta tenui, Halichondria rugosa and Dysidea avara. A wide bacterial diversity was detected according to total genomic DNA-based 16S rDNA clone library, abundant clones with low identify with sequences retrieved from database were found as well as uncultured sponge symbionts. The phylogenetic analysis shows that the bacterial community structure of Stelletta tenui is similar to that of Halichondria rugosa comprising gamma-Proteobacteria and Firmicutes. Whereas, alpha-Proteobacteria, gamma-Protebacteria, Bacteroidetes and uncultured sponge symbionts were found in sponge Dysidea avara, suggesting that Dysidea avara has the highest bacteria diversity among these sponges. A specific sponge–microbe association is suggested based on the difference of bacterial diversity among these three sponges from the same geography location and the observed sponge species-specific bacteria.     

Leptothoe,a new genus of marine cyanobacteria (Synechococcales) and three new species associated with sponges from the Aegean Sea

Cyanobacterial diversity associated with sponges remains underestimated, though it is of great scientific interest in order to understand the ecology and evolutionary history of the symbiotic relationships between the two groups. Of the filamentous cyanobacteria, the genus Leptolyngbya is the most frequently found in association with sponges as well as the largest and obviously polyphyletic group. In this study, five Leptolyngbya‐like sponge‐associated isolates were investigated using a combination of molecular, chemical, and morphological approach and revealed a novel marine genus herein designated Leptothoe gen. nov. In addition, three new species of Leptothoe, Le. sithoniana, Le. kymatousa, and Le. spongobia, are described based on a suite of distinct characters compared to other marine Leptolyngbyaceae species/strains. The three new species, hosted by four sponge species, showed different degrees of host specificity. Leptothoe sithoniana and Le. kymatousa hosted by the sponges Petrosia ficiformis and Chondrilla nucula, respectively, seem to be more specialized than Le. spongobia, which was hosted by the sponges Dysidea avara and Acanthella acuta. All three species contained nitrogen‐fixing genes and may contribute to the nitrogen budget of sponges. Leptothoe spongobia TAU‐MAC 1115 isolated from Acanthella acuta was shown to produce microcystin‐RR indicating that microcystin production among marine cyanobacteria could be more widespread than previously determined.     

The aquiferous systems of three marine demospongiae

The aquiferous systems of three common, coastal, marine Demospongiae, Halichondria panicea (Pallas), Haliclona permollis (Bowerbank) and Microciona Prolifera (Ellis and Solander), are analyzed by measurements of cross-sectional areas of conducting elements. The patterns in demosponges of extremely different organizational morphologies are found to be quantitatively similar. The porocyte nature of the ostia is established for all three species. Choanocyte chamber densities range from 1 to 1.8 × 10⁷ chambers ml⁻¹ with 57 to 95 choanocytes per chamber (means). Cross-sectional area of the intervillar space of the choanocyte collars is calculated to be 12 to 56 times the lateral surface area of the specimen. Velocities of water movement through specific elements of the aquiferous system are calculated from cross-sectional area data and measured oscular flow of Haliclona permollis. The calculated Reynolds numbers lie below the critical value and fluid flow is thus considered laminar throughout the aquiferous systems of these sponges.     

Effects of predation pressure and prey density on short-term indirect interactions between two prey species that share a common predator

1. Generalist predators are important contributors to reliable conservation biological control. Indirect interactions between prey species that share a common generalist predator can influence both community dynamics and the efficacy of biological control. 2. Laboratory cage experiments investigated the impact of the combined consumptive and non-consumptive effects of predation by adult Hippodamia convergens as a shared predator on the population growth and relative abundance of Acyrthosiphon pisum and Aphis gossypii as prey species. Predation pressure and prey density were varied. 3. At low predation pressure the indirect interaction between aphid species was asymmetrical with a proportionally greater negative impact of predation on A. gossypii than on A. pisum. At intermediate predation pressure, the indirect interaction became symmetrical. At high predation pressure and higher levels of prey density, it was asymmetrical with greater negative impact on A. pisum, often driven to local extinction while A. gossypii populations persisted. 4. A linear mixed-effects model including early population growth of both aphid species and predation pressure explained 96% and 92% of the variation in the population growth of A. pisum and A. gossypii, respectively, over an 8-day period. The overall effect of shared predation on the indirect interaction between the two aphid species is best described as apparent commensalism, where A. pisum benefited from early population growth of A. gossypii, while A. gossypii was unaffected by early population growth of A. pisum. Considering these indirect interactions is important for conservation biological control efforts to be successful.     

Functional convergence of microbes associated with temperate marine sponges

Most marine sponges establish a persistent association with a wide array of phylogenetically and physiologically diverse microbes. To date, the role of these symbiotic microbial communities in the metabolism and nutrient cycles of the sponge‐microbe consortium remains largely unknown. We identified and quantified the microbial communities associated with three common Mediterranean sponge species, Dysidea avara, Agelas oroides and Chondrosia reniformis (Demospongiae) that cohabitate coralligenous community. For each sponge we quantified the uptake and release of dissolved organic carbon (DOC) and nitrogen (DON), inorganic nitrogen and phosphate. Low microbial abundance and no evidence for DOC uptake or nitrification were found for D. avara. In contrast A. oroides and C. reniformis showed high microbial abundance (30% and 70% of their tissue occupied by microbes respectively) and both species exhibited high nitrification and high DOC and NH₄⁺ uptake. Surprisingly, these unique metabolic pathways were mediated in each sponge species by a different, and host specific, microbial community. The functional convergence of microbial consortia found in these two sympatric sponge species, suggest that these metabolic processes may be of special relevance to the success of the holobiont.     

Sponge community structure and anti-predator defenses on temperate reefs of the South Atlantic Bight

Predator–prey interactions can play a significant role in shaping the structure of both terrestrial and marine communities. Sponges are major contributors to benthic community structure on temperate reefs and although several studies have investigated how abiotic processes control sponge distributions on these reefs, the role of predation is less clear. We investigated the relationship between sponge predators and the distribution of sponges on temperate reefs in the South Atlantic Bight (SAB), off Georgia, USA. We documented sponge species richness and abundance, spongivorous fish density, and examined the ability of 19 sponge species to chemically and structurally deter predation by fishes. We also conducted reciprocal transplant experiments to determine if predation by fishes contributes to the observed zonation of sponge species on these reefs. Our surveys revealed two distinct sponge assemblages: one characterized by amorphous and encrusting sponge morphotypes colonizing the vertical, rocky outcroppings (scarp sponge community), while the other consisted of pedunculate, digitate, and arborescent growth forms occurring on the sediment-laden reef top (plateau sponge community). Spongivorous fishes were more abundant on the scarp than the plateau and scarp sponges were found to be more effective than plateau sponges at chemically deterring generalist fishes. In contrast, plateau sponges were more reliant on structural defenses: a result consistent with the higher spicule content of their skeletons. Transplant experiments confirmed that predators prevent some plateau sponges from colonizing the scarp even though they possess structural defenses. Thus, predation appears to play a role in shaping sponge community structure on SAB reefs by restricting those species lacking adequate chemical defenses to habitats where there is a paucity of spongivores.     

Polymorphic microsatellite loci in the sponge Crambe crambe (Porifera: Poecilosclerida) and their variation in two distant populations

Seven polymorphic microsatellite loci were characterized in the marine encrusting sponge Crambe crambe from a partial genomic DNA‐enriched library. Preliminary data on allelic variation of these loci in two distant populations of C. crambe are presented to assess their potential utility as high‐resolution genetic markers for this species. The number of alleles per locus ranged between three and 16 and the distributions of allele frequencies differed considerably between the two populations, indicating a marked genetic differentiation between them. These are the first microsatellite loci reported from any species in the phylum.     

Distinct histomorphology for growth arrest and digitate outgrowth in cultivated Haliclona sp. (Porifera: Demospongiae)

The use of sponges in biotechnological processes is limited by the supply problem, and sponge biomass production is becoming a current topic of research. The distinction between characteristics for growth and growth arrest is also important for environmental monitoring. In this study, we analyze the morphology of the digitate outgrowths from the sponge Haliclona sp . The sponge Haliclona sp . was successfully cultivated for 14 months in a closed system. The morphological characterization of growth arrest was performed after submitting explants to starvation‐stress for approximately 2 weeks, to correlate morphology with growth and growth arrest. The digitate outgrowth showed three distinct regions: mature (MR), transition (TR) and immature (IR). Our data suggest a growth developmental program, with collagen fascicles guiding axial growth in IR, followed by progressive development of choanocyte chambers and large aquiferous systems at the more mature proximal region (choanosome). The intercalation of choanocyte chambers and small aquiferous systems inside collagen fascicles previously originated at the IR region can be responsible for thickening expansion and conversion of the collagen fascicles into columnar choanosome in MR. The growth arrest after starvation‐stress assay showed morphological changes in the IR corroborating collagen in the extreme tip of the digitate outgrowth as an important role in guiding of axial growth of Haliclona sp . The identification of distinct morphologies for growth and growth arrest suggest a growth developmental program, and these data could be useful for further investigations addressing sponge biomass gain and environmental monitoring.     

Problems of Coloniality,Modularity, and Individuality in Sponges and Special Features of Their Morphogeneses During Growth and Asexual Reproduction

A comparative analysis of the organization of sponges has been carried out to clarify problems of their coloniality, individuality, and modularity. The morphological, physiological, morphogenetic, and immunological aspects of the problem have been analyzed. The followers of the hypothesis of colonial organization of sponges interpret the process of “new zooid” formation as an “incomplete asexual reproduction.” A comparative analysis of morphogeneses in sponges during growth processes and asexual reproduction has clearly shown them to be different. A rearrangement (remodeling) of structures accompanied by disorganization and reorganization of tissues in neighboring elements of aquiferous system is the basis of growth. Migration of polypotent and secretory cells into the core of bud development is the major mechanism of budding. The formation of new aquiferous units (aquiferous modules) does not represent an “incomplete asexual reproduction.” Thus, the terms “colony” and “zooid” cannot be applied to the sponges. A morphologically separate sponge, irrespective of its level of organization (ascon, sycon, or leucon) and the number of oscula (aquiferous modules) should be considered as an individual.     

Environmental and spatial drivers of beta diversity components of chironomid metacommunities in contrasting freshwater systems

Marine sponges are exposed to predation as well as to a wide array of potentially harmful microorganisms, and therefore they often possess chemical activity against putative predators and/or pathogens. Some crude extracts from sponges are effective in avoiding microbial colonization or potential infections, and in protecting them against predation. Here, the antibacterial activity of 18 sponge species of Antarctic shallow-waters was tested against four Antarctic and four human pathogenic bacteria. Moreover, all sponge extracts were tested for feeding repellence against the seastar Odontaster validus, one of the main predators living in those habitats. All the sponges showed antibacterial activity against at least one bacterial isolate, although not all of them were active against pathogenic bacteria. The antibacterial effect against sympatric bacteria was stronger than to pathogenic bacteria. In contrast, feeding deterrence was low, with similar activities in both hydrophilic and lipophilic extracts. Only four sponges (Myxilla lyssostyla, Phorbas areolatus, Polymastia invaginata and Iophon sp.), presented repellent chemical defenses. Therefore, we conclude that chemical defenses are widespread in Antarctic shallow-water sponges, and in fact, these sponges are better protected against bacteria than against the seastar predator. We conclude that Antarctic sponges represent a valuable source of biological active compounds with pharmacological and potential ecological relevance.     

Vegetation structure influences predation rates of early nests in subarctic breeding waders

Ground-nesting species are vulnerable to a wide range of predators and often experience very high levels of nest predation. Strategies to reduce nest vulnerability can include concealing nests in vegetation and/or nesting in locations in which nests and eggs are camouflaged and less easy for predators to locate. These strategies could have important implications for the distribution of ground-nesting species and the success rates of nests in areas with differing vegetation structure. However, the factors influencing the success of nest concealment and camouflage strategies in ground-nesting species are complex. Here we explore the effects of local vegetation structure and extent of nest concealment on nest predation rates in a range of ground-nesting, sympatric wader species with differing nest concealment strategies (open-nest species: Oystercatcher Haematopus ostralegus, Golden Plover Pluvialis apricaria and Whimbrel Numenius phaeopus; concealed-nest species: Black-tailed Godwit Limosa limosa, Redshank Tringa totanus and Snipe Gallinago gallinago) in south Iceland, in landscapes that comprise substantial variability in vegetation structure at a range of scales. We monitored 469 nests of these six wader species in 2015 and 2016 and ~40% of these nests were predated. Nest predation rates were similar for open-nest and concealed-nest species and did not vary with vegetation structure in the surrounding landscape, but nest-concealing species were ~10% more likely to have nests predated when they were poorly concealed, and the frequency of poorly concealed nests was higher in colder conditions at the start of the breeding season. For concealed-nest species, the reduced capacity to hide nests in colder conditions is likely to reflect low rates of vegetation growth in such conditions. The ongoing trend for warmer springs at subarctic latitudes could result in more rapid vegetation growth, with consequent increases in the success rates of early nests of concealed-nest species. Temperature-related effects on nest concealment from predators could thus be an important mechanism through which climate change affecting vegetation could have population-level impacts on breeding birds at higher latitudes.     

Sponge-Cell Culture? A Molecular Identification Method for Sponge Cells

Dissociated sponge cells are easily confused with unicellular organisms. This has been an obstacle in the development of sponge-cell lines. We developed a molecular detection method to identify cells of the sponge Dysidea avara in dissociated cell cultures. The 18S ribosomal RNA gene from a Dysidea avara specimen was sequenced and compared to eukaryotic 18S rDNA sequences picked up from a proliferating cell culture that originated from a dissociated Dysidea avara specimen. Our method proved unambiguously that this was not a sponge-cell culture. Therefore, it provides a valuable tool for further research on sponge-cell cultures.     

Bacterial community diversity associated with four marine sponges from the South China Sea based on 16S rDNA-DGGE fingerprinting

Culture-independent 16S rDNA-DGGE fingerprinting and phylogenetic analysis were used to reveal the community structure and diversity of the predominant bacteria associated with the four sponges Stelletta tenui, Halichrondria, Dysidea avara, and Craniella australiensis from the South China Sea for the first time. Sponge total community DNA extracted with a direct grinding disruption based method was used successfully after series dilution for 16S rDNA PCR amplification, which simplifies the current procedure and results in good DGGE banding profiles. 16S rDNA-V3 fragments from 42 individual DGGE bands were sequenced and the detailed corresponding bacteria were found in sponges for the fist time based on BLAST results. The sponge-associated bacteria are sponge host-specific because each of the tested four sponges from the same geographical location has different predominant bacterial diversity. Proteobacteria, e.g. α, β and γ subdivisions, make up the majority of the predominant bacteria in sponges and are perhaps in close symbiotic relationship with sponges. Though similar bacteria with close phylogenetic relationships were found among different sponges, the sponge-associated predominant bacterial community structures differ. Sponge C. australiensis has the greatest bacterial diversity, with the four bacteria phyla Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria, followed by the sponge D. avara with the two phyla Proteobacteria and Bacteroidetes, and the sponges S. tenui and Halichrondria with the phylum Proteobacteria. DGGE fingerprint-based analysis should ideally be integrated with band cloning and sequencing, phylogenetic analysis and molecular techniques to obtain precise results in terms of the microbial community and diversity.     

Sponge cell aggregation: checkpoints in development indicate a high level of organismal complexity

Studies of regeneration provide insight across many scales of animal biology from the processes of cellular communication to the ecology of whole populations. Sponges are highly regenerative animals, with studies showing adults can both recover large portions of their body after predation or damage due to storms, and even reform whole individuals, via an aggregation stage, from dissociated tissues. While sponges are clearly highly regenerative, few studies actually show dissociated cells forming functional individuals. As sponges often serve as model organisms for studying the development and function of traits in metazoans, determining the universality and mechanics of their regeneration potential is important. We tested the capacity of members of seven sponge species from temperate freshwater and marine environments, from a range of taxonomic positions, and with different habits, to form functional sponges after dissociation. Development to a functional sponge progressed through a series of checkpoints: the sorting of cells and removal of debris; adhesion to a substrate and differentiation of cells; organization of cells into tissues; and regionalization of tissues. Two of the seven species tested, Spongilla lacustris and Haliclona cf. permollis, progressed through all four checkpoints, while the remaining five species progressed to various levels of development before aggregates disintegrated. Our findings highlight three important conclusions: (1) The ability of aggregates to differentiate into functional sponges is not as widespread as previously thought; (2) The species‐specific ability of aggregates to develop to functional sponges appears to be an adaptive trait; and (3) The progression of development in aggregates through checkpoints, which in later development involves formation of tissues and regionalization of tissues, highlights the complexity of the sponge body plan and suggests fundamental rules in development shared across metazoans.     

Pelagic larvae of New England intertidal gastropods II. Anachis avara

Summary The egg case, pelagic larvae, and young juvenile of Anachis avara Say, a species of prosobranch gastropod belonging to the family Columbellidae, are described and figured from specimes reared in the laboratory.

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Zusammenfassung Die Arbeit beschreibt die Eierkapseln, die pelagischen Larven und die Jugendformen von Anachis avara Say, einer prosobranchen Gastropoden-Spezies, die zur Familie der Columbelliden gehört. Abbildungen von Exemplaren, die im Laboratorium aufgezogen wurden, werden wiedergegeben.

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Contribution number 1341, Woods Hole Oceanographic Institution. This research was supported by N.S.F. grant no. 17883.