Sponge Aplysina Research Articles

Synthesis of nanostructured chitin–hematite composites under extreme biomimetic conditions

Chitin of poriferan origin is a unique and thermostable biological material. It also represents an example of a renewable materials source due to the high regeneration ability of Aplysina sponges under marine ranching conditions. Chitinous scaffolds isolated from the skeleton of the marine sponge Aplysina aerophoba were used as a template for the in vitro formation of Fe2O3 under conditions (pH ∼ 1.5, 90 °C) which are extreme for biological materials. Novel chitin–Fe2O3 three dimensional composites, which have been prepared for the first time using hydrothermal synthesis, were thoroughly characterized using numerous analytical methods including Raman spectroscopy, XPS, XRD, electron diffraction and HR-TEM. We demonstrate the growth of uniform Fe2O3 nanocrystals into the nanostructured chitin substrate and propose a possible mechanism of chitin–hematite interactions. Moreover, we show that composites made of sponge chitin–Fe2O3 hybrid materials with active carbon can be successfully used as electrode materials for electrochemical capacitors.

Complex ecological associations: competition and facilitation in a sponge–algal interaction

Over the past few decades, Caribbean coral reefs have undergone a phase shift from coral-dominated to algal-dominated communities due to several factors, including increased input of anthropogenic nutrients. With the decline in coral cover, sponges have also become more dom- inant members of Caribbean coral reef communities. Increased algal and sponge dominance on Caribbean reefs has led to an increase in the frequency of interaction between these 2 groups. This study used a factorial design to assess the independent and interactive effects of contact and elevated nutrient levels on 2 common members of these communities, the sponge Aplysina cauli- formis, and the macroalga Microdictyon marinum. Algal contact had a significant negative phys- iological effect on A. cauliformis, affecting both the host sponge and its cyanobacterial symbionts. While elevated nutrient levels had some positive effects on the sponge photosymbionts, this only occurred in the absence of algal contact or a shading/abrasion control, and elevated nutrient lev- els had a negative effect on the sponge holobiont. In contrast, M. marinum responded positively to experimentally enhanced nutrient levels and to sponge contact under ambient nutrient regimes, but was not affected by sponge contact under elevated nutrient concentrations. Stable isotope enrichment experiments showed that the alga's positive response to sponge contact was associated with nitrogen transfer from the sponge over the course of the experiment. Thus, while A. cauliformis facilitates increased productivity in M. marinum, algal contact competitively inhibits sponge condition.

Antifouling activity of symbiotic bacteria from sponge Aplysina gerardogreeni

A key area in marine antifoulant research is the discovery of new environmentally friendly solutions that prevent biofilm formation and associated biocorrosion. Taking into consideration the natural mechanisms of marine organisms to protect against epibiosis, new biomimetic solutions can be utilised against biofouling, and marine bacteria are promising agents. Therefore, the goal of this study was to identify cultivable bacteria with antifouling (AF) activity associated with the sponge Aplysina gerardogreeni. A collection of 63 bacteria was isolated, and the organic extracts were assayed against various microfouler strains (16 bacteria and five microalgae). The results showed that 87% of bacterial extracts were active against the microfoulers tested. Sixteen of them can be considered to possess AF potential and belong to the genera Bacillus, Micrococcus, Paracoccus, Pseudobacter, Pseudovibrio, Psychrobacter, Staphylocuccus and Terribacillus. Bioactivity showed temporal variations; the highest activity was in February and June and the lowest in October. Bacillus bacteria were dominant and showed AF activities throughout the year. The results revealed those marine bacteria sponge-associated and the genus Bacillus in particular, are promising AF agents.

Rubrobacter aplysinae sp. nov., isolated from the marine sponge Aplysina aerophoba

A Gram-stain-positive, non-spore-forming bacterium (strain RV113(T)) was isolated from the marine sponge Aplysina aerophoba. 16S rRNA gene sequence analysis showed that strain RV113(T) belongs to the genus Rubrobacter, and is related most closely to Rubrobacter bracarensis VF70612_S1(T) (96.9% similarity) and more distantly related (<93%) to all other species of the genus Rubrobacter. The peptidoglycan diamino acid was lysine. Strain RV113(T) exhibited a quinone system with menaquinone MK-8 as the predominant compound. The polar lipid profile of strain RV113(T) consisted of the major compounds phosphatidylglycerol and two unidentified phosphoglycolipids. The major fatty acid was anteiso-C17 : 0ω9c. These chemotaxonomic traits are in agreement with those of other species of the genus Rubrobacter. The results of physiological and biochemical tests allowed the clear phenotypic differentiation of strain RV113(T) from all recognized Rubrobacter species. Strain RV113(T) is thus considered to represent a novel species, for which the name Rubrobacter aplysinae sp. nov. is proposed. The type strain is RV113(T) ( = DSM 27440(T) = CECT 8425(T)).

Exploring Individual- to Population-Level Impacts of Disease on Coral Reef Sponges: Using Spatial Analysis to Assess the Fate, Dynamics, and Transmission of Aplysina Red Band Syndrome (ARBS)

BackgroundMarine diseases are of increasing concern for coral reef ecosystems, but often their causes, dynamics and impacts are unknown. The current study investigated the epidemiology of Aplysina Red Band Syndrome (ARBS), a disease affecting the Caribbean sponge Aplysina cauliformis, at both the individual and population levels. The fates of marked healthy and ARBS-infected sponges were examined over the course of a year. Population-level impacts and transmission mechanisms of ARBS were investigated by monitoring two populations of A. cauliformis over a three year period using digital photography and diver-collected data, and analyzing these data with GIS techniques of spatial analysis. In this study, three commonly used spatial statistics (Ripley’s K, Getis-Ord General G, and Moran’s Index) were compared to each other and with direct measurements of individual interactions using join-counts, to determine the ideal method for investigating disease dynamics and transmission mechanisms in this system. During the study period, Hurricane Irene directly impacted these populations, providing an opportunity to assess potential storm effects on A. cauliformis and ARBS.ResultsInfection with ARBS caused increased loss of healthy sponge tissue over time and a higher likelihood of individual mortality. Hurricane Irene had a dramatic effect on A. cauliformis populations by greatly reducing sponge biomass on the reef, especially among diseased individuals. Spatial analysis showed that direct contact between A. cauliformis individuals was the likely transmission mechanism for ARBS within a population, evidenced by a significantly higher number of contact-joins between diseased sponges compared to random. Of the spatial statistics compared, the Moran’s Index best represented true connections between diseased sponges in the survey area. This study showed that spatial analysis can be a powerful tool for investigating disease dynamics and transmission in a coral reef ecosystem.

Molecular community profiling reveals impacts of time, space, and disease status on the bacterial community associated with the Caribbean spongeAplysina cauliformis

Reports of marine sponge diseases have increased in recent years, but few etiologic agents have been identified. Aplysina red band syndrome (ARBS), a condition observed in the Caribbean sponge Aplysina cauliformis, is characterized by a rust-colored leading margin. Culture-independent methods (terminal restriction fragment length polymorphism and clone library analyses) were used to assess bacterial communities associated with healthy and ARBS-affected sponges from two locations over 2 years. Although the bacterial communities associated with healthy and ARBS-affected sponges were significantly different, the sponges maintained a core bacterial community across space, time, and health status. Ten terminal restriction fragments were shown to change significantly between sponge health conditions, with six increasing in abundance with disease and four decreasing. The prevalence of the photosymbiont Synechococcus spongiarum decreased with ARBS infection, suggesting a functional consequence of disease. After cultivating a red-pigmented Leptolyngbya strain from ARBS lesions, transmission studies were conducted to determine whether this organism was the ARBS pathogen. Despite significantly increased abundance of Leptolyngbya spp. in diseased sponges, signs of ARBS were not observed in healthy sponges following 24 days of contact with the cultured strain. Additional work with this model system is needed to increase our understanding of the dynamics of marine diseases.

An aeroplysinin-1 specific nitrile hydratase isolated from the marine sponge Aplysina cavernicola.

A nitrile hydratase (NHase) that specifically accepts the nitrile aeroplysinin-1 (1) as a substrate and converts it into the dienone amide verongiaquinol (7) was isolated, partially purified and characterized from the Mediterranean sponge Aplysina cavernicola; although it is currently not known whether the enzyme is of sponge origin or produced by its symbiotic microorganisms. The formation of aeroplysinin-1 and of the corresponding dienone amide is part of the chemical defence system of A. cavernicola. The latter two compounds that show strong antibiotic activity originate from brominated isoxazoline alkaloids that are thought to protect the sponges from invasion of bacterial pathogens. The sponge was shown to contain at least two NHases as two excised protein bands from a non denaturating Blue Native gel showed nitrile hydratase activity, which was not observed for control samples. The enzymes were shown to be manganese dependent, although cobalt and nickel ions were also able to recover the activity of the nitrile hydratases. The temperature and pH optimum of the studied enzymes were found at 41 °C and pH 7.8. The enzymes showed high substrate specificity towards the physiological substrate aeroplysinin-1 (1) since none of the substrate analogues that were prepared either by partial or by total synthesis were converted in an in vitro assay. Moreover de-novo sequencing by mass spectrometry was employed to obtain information about the primary structure of the studied NHases, which did not reveal any homology to known NHases.

Poriferan chitin as a template for hydrothermal zirconia deposition

Chitin is a thermostable biopolymer found in various inorganic-organic skeletal structures of numerous invertebrates including sponges (Porifera). The occurrence of chitin within calcium- and silica-based biominerals in organisms living in extreme natural conditions has inspired development of new (extreme biomimetic) synthesis route of chitin-based hybrid materials in vitro. Here, we show for the first time that 3D-α-chitin scaffolds isolated from skeletons of the marine sponge Aplysina aerophoba can be effectively mineralized under hydrothermal conditions (150°C) using ammonium zirconium(IV) carbonate as a precursor of zirconia. Obtained chitin-ZrO2 hybrid materials were characterized by FT-IR, SEM, HRTEM, as well as light and confocal laser microscopy. We suggest that formation of chitin-ZrO2 hybrids occurs due to hydrogen bonds between chitin and ZrO2.

Polyketide genes in the marine sponge Plakortis simplex: a new group of mono‐modular type I polyketide synthases from sponge symbionts

SummarySponge symbionts are a largely unexplored source of new and unusual metabolic pathways. Insights into the distribution and function of metabolic genes of sponge symbionts are crucial to dissect and exploit their biotechnological potential. Screening of the metagenome of the marine sponge Plakortis simplex led to the discovery of the swf family, a new group of mono-modular type I polyketide synthase/fatty acid synthase (PKS/FAS) specifically associated with sponge symbionts. Two different examples of the swf cluster were present in the metagenome of P. simplex. A third example of the cluster is present in the previously sequenced genome of a poribacterium from the sponge Aplysina aerophoba but was formerly considered orthologous to the wcb/rkp cluster. The swf cluster was also found in six additional species of sponges. Therefore, the swf cluster represents the second group of mono-modular PKS, after the supA family, to be widespread in marine sponges.The putative swf operon consists of swfA (type I PKS/FAS), swfB (reductase and sulphotransferase domains) and swfC (radical S-adenosylmethionine, or radical SAM). Activation of the acyl carrier protein (ACP) domain of the SwfA protein to its holo-form by co-expression with Svp is the first functional proof of swf type genes in marine sponges. However, the precise biosynthetic role of the swf clusters remains unknown.

Minor Brominated Compounds from Extract of the Sponge Aplysina sp.

Previously unknown minor brominated compounds 1a, 1b, and 2 were isolated from the EtOH extract of the sponge Aplysina sp. (South China Sea). Their structures were established by NMR spectroscopic methods and mass spectrometry.

Species of the genus Periclimenaeus (Decapoda, Caridea, Palaemonidae) associated with sponges from the Veracruz Coral Reef System National Park, SW Gulf of Mexico

The relationship between the depth of the carapace of shrimps of the genus Periclimenaeus and canal diameter of the sponges Ailochroia crassa, Aplysina fistularis, Callyspongia armigera, Ircinia fistularis, I. strobilina and Sidonops neptuni from Veracruz Coral Reef System National Park, SW Gulf of Mexico, Mexico, were studied. Thirty sponges were collected manually, using SCUBA, and placed underwater in hermetically sealed plastic bags, 1 : 1 formalin/alcohol solution was added to the plastic bags; then the crustaceans were manually removed and fixed in 70% ethanol. Correlations between mean depth of the carapace of a shrimp species and canal diameter of the sponges inhabited by that shrimp species were estimated. The total number of Periclimenaeus shrimps was 108, with three species associated with six species of sponges. The shrimps were found on seven reefs of the coral reef system. The most abundant species of shrimp were P. perlatus and P. wilsoni with 67 and 35 individuals, respectively. In the association of the shrimps with the sponges, P. perlatus was found in 84% of Aplysina fistularis specimens and P. wilsoni was found in 66% of Ircinia fistularis specimens. The deepest carapace was found in the females of P. pearsei with 3.96 ± 0.04 mm. The regression of the depth of the carapace of the shrimps against the canal diameter of the species of sponges showed a positive and significant relationship (P<0.001). These correlations raise the need to explore the full range of morphological and physiological adaptations of species of the genus Periclimenaeus to the sponges they inhabit, and the geographical distribution of Periclimenaeus-sponge interactions is essential to understand how the use of sponge hosts has contributed to the biodiversity of shrimps throughout the SW Gulf of Mexico.

Extreme Biomimetics: formation of zirconium dioxide nanophase using chitinous scaffolds under hydrothermal conditions.

Chitinous scaffolds isolated from the skeleton of marine sponge Aplysina cauliformis were used as a template for the in vitro formation of zirconium dioxide nanophase from ammonium zirconium(iv) carbonate (AZC) under extreme conditions (150 °C). These novel zirconia-chitin based composites were prepared for the first time using hydrothermal synthesis, and were thoroughly characterized using a plethora of analytical methods. The thermostability of the chitinous 3D matrix makes it ideal for use in the hydrothermal synthesis of monoclinic nanostructured zirconium dioxide from precursors like AZC. These zirconium-chitin composites have a high potential for use in a broad range of applications ranging from synthetic catalysis to biocompatible materials for bone and dental repair. The synthetic methods presented in this work show an attractive route for producing monoclinic zirconium dioxide on a 3D biocompatible scaffold with ease.

Diversity of the candidate phylum Poribacteria in the marine sponge Aplysina fulva

Poribacterial clone libraries constructed for Aplysina fulva sponge specimens were analysed with respect to diversity and phylogeny. Results imply the coexistence of several, prevalently “intra-specific” poribacterial genotypes in a single sponge host, and suggest quantitative analysis as a desirable approach in studies of the diversity and distribution of poribacterial cohorts in marine sponges.

The Antiangiogenic Compound Aeroplysinin-1 Induces Apoptosis in Endothelial Cells by Activating the Mitochondrial Pathway

Aeroplysinin-1 is a brominated metabolite extracted from the marine sponge Aplysina aerophoba that has been previously characterized by our group as a potent antiangiogenic compound in vitro and in vivo. In this work, we provide evidence of a selective induction of apoptosis by aeroplysinin-1 in endothelial cells. Studies on the nuclear morphology of treated cells revealed that aeroplysinin-1 induces chromatin condensation and nuclear fragmentation, and it increases the percentage of cells with sub-diploid DNA content in endothelial, but not in HCT-116, human colon carcinoma and HT-1080 human fibrosarcoma cells. Treatment of endothelial cells with aeroplysinin-1 induces activation of caspases-2, -3, -8 and -9, as well as the cleavage of apoptotic substrates, such as poly (ADP-ribose) polymerase and lamin-A in a caspase-dependent mechanism. Our data indicate a relevant role of the mitochondria in the apoptogenic activity of this compound. The observation that aeroplysinin-1 prevents the phosphorylation of Bad relates to the mitochondria-mediated induction of apoptosis by this compound.

Disease and nutrient enrichment as potential stressors on the Caribbean sponge Aplysina cauliformis and its bacterial symbionts

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 456:101-111 (2012) - DOI: https://doi.org/10.3354/meps09716 Disease and nutrient enrichment as potential stressors on the Caribbean sponge Aplysina cauliformis and its bacterial symbionts D. J. Gochfeld1,2,*, C. G. Easson2, C. J. Freeman3, R. W. Thacker3, J. B. Olson4 1National Center for Natural Products Research and 2Environmental Toxicology Research Program, University of Mississippi, University, Mississippi 38677-1848, USA 3Department of Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA 4Department of Biological Sciences, University of Alabama, Tuscaloosa, Alabama 35487, USA *Email: gochfeld@olemiss.edu ABSTRACT: Sponge diseases have recently emerged as potential forces structuring coral reefs. The increasing prevalence of disease on reefs may be due to changes in the virulence of pathogens and/or to decreases in host resistance as a result of changing environmental conditions. Coral reef ecosystems typically thrive in oligotrophic waters; however, runoff of fertilizers or sewage that contains elevated concentrations of nutrients can lead to eutrophic conditions. Aplysina cauliformis is a dominant member of the Caribbean sponge community, and is susceptible to Aplysina Red Band Syndrome (ARBS), a disease that causes reduced sponge growth and survival. We assessed the independent and interacting effects of nutrient enrichment and disease on A. cauliformis, using a factorial field experiment in which healthy and diseased sponges were exposed to nutrient-enriched or non-enriched treatments. Impacts on ARBS virulence (rate of lesion growth) and host response (both sponge and cyanobacterial symbiont growth and physiology) were assessed. ARBS lesions increased rapidly regardless of nutrient treatment, and disease had a significantly greater detrimental impact on sponges than did nutrient enrichment, as evidenced by a decline in sponge mass and reduced total protein content. The sponge−cyanobacterial symbiont relationship was less impacted by disease, although the sponge-associated bacterial community was significantly affected by sponge condition, with healthy sponges and diseased tissue hosting significantly different bacterial assemblages. In contrast, nutrient enrichment had no effects on sponge or symbiont physiology. Disease is a much greater stressor than eutrophication on the growth and physiology of A. cauliformis and its cyanobacterial symbionts. KEY WORDS: Sponge disease · Nutrient enrichment · Symbiosis · Stressors Full text in pdf format PreviousNextCite this article as: Gochfeld DJ, Easson CG, Freeman CJ, Thacker RW, Olson JB (2012) Disease and nutrient enrichment as potential stressors on the Caribbean sponge Aplysina cauliformis and its bacterial symbionts. Mar Ecol Prog Ser 456:101-111. https://doi.org/10.3354/meps09716 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 456. Online publication date: June 07, 2012 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2012 Inter-Research.

Diversity of Nonribosomal Peptide Synthetase Genes in the Microbial Metagenomes of Marine Sponges

Genomic mining revealed one major nonribosomal peptide synthetase (NRPS) phylogenetic cluster in 12 marine sponge species, one ascidian, an actinobacterial isolate and seawater. Phylogenetic analysis predicts its taxonomic affiliation to the actinomycetes and hydroxy-phenyl-glycine as a likely substrate. Additionally, a phylogenetically distinct NRPS gene cluster was discovered in the microbial metagenome of the sponge Aplysina aerophoba, which shows highest similarities to NRPS genes that were previously assigned, by ways of single cell genomics, to a Chloroflexi sponge symbiont. Genomic mining studies such as the one presented here for NRPS genes, contribute to on-going efforts to characterize the genomic potential of sponge-associated microbiota for secondary metabolite biosynthesis.

11-Oxoaerothionin isolated from the marine sponge Aplysina fistularis shows anti-inflammatory activity in LPS-stimulated macrophages

Marine sponges of the order Verongida are a rich source of biologically active bromotyrosine-derived secondary metabolites. However, none of these compounds are known to display anti-inflammatory activity. In the present investigation, we report the anti-inflammatory effects of 11-oxoaerothionin isolated from the Verongida sponge Aplysina fistularis. When RAW264.7 cells and primary macrophages were preincubated with 11-oxoaerothionin and stimulated with LPS (lipopolysaccharide), a concentration-dependent inhibition of iNOS(inducible nitric oxide synthase) protein and NO-2 (Nitrite) production were observed. The same effect was observed when proinflammatory cytokines and PGE2 (Prostaglandin E2) production was evaluated. In summary, we demonstrated that in the presence of LPS, 11-oxoaerothionin suppresses NO2 and iNOS expression as well as inflammatory cytokines and PGE2.

Trade-Offs in Defensive Metabolite Production But Not Ecological Function in Healthy and Diseased Sponges

Diseases of marine organisms, and sponges in particular, are increasingly reported worldwide. Prior research indicates that the survival of sponges on reefs is due largely to their production of biologically active secondary metabolites that provide protection from a diversity of stressors. Aplysina Red Band Syndrome (ARBS) is an emerging disease affecting Caribbean rope sponges (Aplysina spp.), but it is not known whether secondary metabolites play a role in disease susceptibility and resistance. To investigate whether differences in secondary metabolites may explain variability in susceptibility to ARBS in Aplysina cauliformis, we used high performance liquid chromatography (HPLC) to generate chemical profiles from healthy tissue in both healthy and diseased sponges, and quantified peak areas for 15 metabolites. Analyses of healthy and diseased sponges revealed qualitative and quantitative differences in their chemical profiles. Aplysamine-1 and fistularin-3 were produced in significantly higher concentrations by healthy sponges, whereas aerothionin and 11-oxoaerothionin were found only in diseased sponges. At natural concentrations, extracts from both healthy and diseased sponges deterred feeding by an omnivorous reef fish. Fistularin-3 deterred feeding at concentrations found in healthy sponges, but not at concentrations found in diseased sponges. Aerothionin deterred feeding at concentrations found in diseased sponges, and may at least partially replace the loss of fistularin-3 as a feeding deterrent compound following pathogenesis, suggesting a trade-off in the production of feeding deterrent compounds. Extracts from healthy and diseased sponges inhibited bacterial growth, and both aplysamine-1 and fistularin-3 displayed selective antibacterial activity. Despite differences in secondary metabolite production between healthy and diseased sponges, the stress associated with ARBS does not appear to compromise the ability of A. cauliformis to maintain defenses against some of its natural enemies.

Temporal Trends in the Secondary Metabolite Production of the Sponge Aplysina aerophoba

Temporal changes in the production of secondary metabolites are far from being fully understood. Our study quantified, over a two-year period, the concentrations of brominated alkaloids in the ectosome and the choanosome of Aplysina aerophoba, and examined the temporal patterns of these natural products. Based on standard curves, we quantified the concentrations of aerophobin-2, aplysinamisin-1, and isofistularin-3: three of the four major peaks obtained through chemical profiling with high-performance liquid chromatography. Our results showed a striking variation in compound abundance between the outer and inner layers of the sponge. The ectosome showed high concentrations of bromocompounds during the summer months, while the choanosome followed no pattern. Additionally, we found that, from the outer layer of the sponge, aerophobin-2 and isofistularin-3 were significantly correlated with water temperature. The present study is one of the first to document quantitative seasonal variations in individual compounds over multiple years. Further studies will clarify the role of environmental, biological, and physiological factors in determining the seasonal patterns in the concentration of brominated alkaloids.

Aplysfistularine: a novel dibromotyrosine derivative isolated from Aplysina fistularis

The new dibromotyrosine derivative 3,5-dibromo-4-[3'dimethylamonium]propoxyphenyl]-N,N,N-trimethylethanamonium, here referred to as aplysfistularine (1), was isolated from the marine sponge Aplysina fistularis along with 2-(3,5-dibromo-4methoxyphenyl)-N,N,N-trimethylethanamonium (2), aplysterol (3) and 24,28-didehydroaplysterol (4). Their identification was performed by mass spectrometry, infrared, 1H and 13C NMR, and by comparison with literature data. Compound 2 and the mixture of 3 and 4 were tested in vitro (inhibitory activity) with supercoiled DNA relaxation techniques, and showed inhibitory activity on human DNA topoisomerase II-α. Compound 1 was not tested due to paucity of the material.