Antibacterial Activity and Drug Release of Chitosan Sponge Containing Doxycycline Hyclate

The purpose of the present study was to develop and characterize the chitosan sponges loading with doxycycline hyclate and their antibacterial activities. The pore density of chitosan sponge prepared with freeze drying technique was increased as the higher concentrated chitosan solution was used. The sponge prepared from 10% w/w of the chitosan solution and crosslinking with glutaraldehyde solution was utilized for loading with doxycycline hyclate. The drug release and sustainable antibacterial activity of fabricated sponge were assessed using dissolution test and agar diffusion test, respectively. Drug release from non-crosslinked sponge into phosphate buffer pH7.4 was slower than that from crosslinked sponge since the former could absorb the medium and form gel to retard the initial drug diffusion. Sustainable antibacterial activity of developed sponge was evident against S. aureus and E. coli. In conclusion, the in vitro release profile and antibacterial efficiency indicated that doxycycline hyclate could be sustained using chitosan sponge.     

Bacteria associated with the sponge Spongia officinalis as indicators of contamination

A lot of micro-organisms can been found in the sponge body. In the present study, bacteria and fungi were isolated from the massive commercial sponge Spongia officinalis Linnaeus, 1759 (Porifera, Demospongiae). Samples (sponge extract and sea water) were collected from several areas of the Aegean Sea (ports, beaches, eutrophic gulf and open sea) with different levels and types of contamination. Bacteria identified in sponge extract belong to the genera Escherichia, Morganella, Proteus, Pasteurella, Aeromonas, Pseudomonas and Acinetobacter. Two fungal genera, Trichosporum and Fisarium, were identified in sponge extract. Bacterial load was greater in sponge extract than in proximal sea water, reflecting the sponge ability to concentrate bacteria in its body. The assessment of bacteria associated with the sponge S. officinalis as indicators of contamination is proposed.     

Sponge–seaweed associations in species of Ptilophora (Gelidiaceae, Rhodophyta)

Sponge–seaweed associations in the seaweed genus Ptilophora are poorly understood; therefore, 94 specimens, representing all 17 species of Ptilophora, were examined to detail this phenomenon. All but 2 Ptilophora species were shown to produce surface proliferations, with 13 species found to have sponge associations. Evidence for facultative sponge epiphytism was found with species–specific interactions being unlikely. Results show that surface proliferations are not induced by sponge epiphytes, as they often occur in the absence of sponge epiphytes, and vice versa. The significant number of proliferate thalli found with sponge epiphytes suggests that there is a likely relationship between the presence of surface proliferations and sponge infestation. Sponge epiphytes and Ptilophora species appeared structurally related in that the sponge probably exploits a niche habitat provided by the alga, for which surface proliferations might aid the sponge in bonding to the alga.     

Compositional analysis of archaeal communities in high and low microbial abundance sponges in the Misool coral reef system,Indonesia

The high/low microbial abundance (HMA/LMA) dichotomy in sponges has been the subject of several studies over recent years, but few studies have analysed this dichotomy in terms of the sponge archaeal community and function. Using a 16S rRNA gene barcoded pyrosequencing approach and predictive functional analysis (PICRUSt) we compared the archaeal composition, richness and predicted function of one HMA sponge (Xestospongia testudinaria), one LMA sponge (Stylissa carteri) and one sponge species of unknown microbial abundance (Aaptos lobata). Although most of the archaeal sequences were assigned to the Crenarchaeota phylum, S. carteri had the highest percentage of sequences assigned to the Euryarchaeota phylum. Variation among sponge species explained >85% of the variation in archaeal operational taxonomic unit (OTU) composition with each sponge species forming a distinct cluster. There were significant differences in predicted PICRUSt profiles among sponge species, suggesting that archaeal communities present in the studied sponge species may perform different functions. X. testudinaria and A. lobata were similar both in terms of OTU and KEGG orthologues composition, which may indicate that A. lobata is a HMA sponge. Additionally, some of the most enriched functions seem to be related to traits associated with high and low microbial abundance sponges.     

Mangrove-sponge associations: a possible role for tannins

A positive correlation between sponge coverage and tannin concentrations in prop roots of Rhizophora mangle L. has previously been reported. However, the ecological role of tannins within the mangrove sponge association remains speculative. This study investigated whether tannins play a role in sponge recruitment and assessed tannin and polyphenol production in R. mangle roots in response to sponge colonization. We demonstrated in a field experiment using artificial substrates with different tannin concentrations that tannins are positively involved in larval recruitment of the sponge Tedania ignis and that roots significantly enhanced tannin and polyphenolic content in response to natural and experimental sponge fouling. Differential recruitment in response to tannins may have been the result of a behavioral response in sponge larvae. It is also possible that tannins affected the structure of the fouling microbial biofilm on the artificial substrate, or tannins affected the post-settlement dynamics of sponge recruits. Elevations in concentrations of tannins and polyphenolic compounds upon coverage with sponges, combined with differential recruitment of T. ignis in response to differences in tannin concentrations, may indicate a positive feedback in recruitment. This may in part explain the typical heterogeneity in sponge coverage and community composition among roots.     

Invasive alga <Emphasis Type="Italic">Caulerpa racemosa</Emphasis> var. <Emphasis Type="Italic">cylindracea</Emphasis> makes a strong impact on the Mediterranean sponge <Emphasis Type="Italic">Sarcotragus spinosulus</Emphasis>

Here we present the first observation of the impact of the invasive Caulerpa racemosa var. cylindracea on native photophilic sponge species in the Adriatic Sea, with special focus on Sarcotragus spinosulus. Caulerpa racemosa var. cylindracea is able to completely overgrow the sponge, developing an exceptionally thick canopy with a maximum measured density of 1,887 m of stolons m⁻² and 40,561 fronds m⁻². Necrosis of the sponge surface was significantly correlated with the algal dry biomass, frond number and stolon length. Dense algal canopy, penetration of the algal stolon and rhizoids into the sponge oscula and covering of the ostiae probably diminishes the seawater circulation through the sponge and consequently results in its smothering and even death. We suggest that chemotropism is the reason why C. racemosa penetrates the sponge oscula and establishes such dense canopy on the sponge.     

Detection and Phylogenetic Analysis of Novel Crenarchaeote and Euryarchaeote 16S Ribosomal RNA Gene Sequences from a Great Barrier Reef Sponge

The presence of Archaea in the Great Barrier Reef marine sponge Rhopaloeides odorabile was investigated by 16S ribosomal RNA community analysis of total DNA extracted from the sponge tissue. The 16S rRNA gene sequences corresponding to group I crenarchaeotes and group II euryarchaeotes were recovered from R. odorabile tissue. The location of archaeal cells within the sponge tissue was investigated using fluorescently labeled oligonucleotide probes. The presence of Archaea was confirmed within all regions of the sponge tissue from R. odorabile, with a significantly higher number of archaeal cells located in the pinacoderm than the mesohyl region. This is the first report of euryarchaeaotes associated with marine sponges. Received April 16, 2001; accepted June 27, 2001     

The effects of cadmium and mercury on gemmule formation and gemmosclere morphology in Ephydatia fluviatilis (Porifera : Spongillidae)

A laboratory study of the effects of cadmium and mercury on the freshwater sponge, Ephydatia fluviatilis, was conducted. Sponge cuttings were exposed to concentrations of cadmium or mercury which ranged from 1.000 to 0.001 ppm for one month. The responses exhibited by the specimens resulted in four groups characterized as follows: sponge colony survived and produced gemmules with normal gemmoscleres; sponge colony survived and produced gemmules with malformed gemmoscleres; sponge colony survived but did not gemmulate; sponge colony died. A direct discriminant functions analysis of the four sponge groups, metal concentrations and other chemical data established a highly significant correlation between increasing metal concentrations and amount of damage to the sponge.     

Skeletal response of <Emphasis Type="Italic">Lophelia pertusa</Emphasis> (Scleractinia) to bioeroding sponge infestation visualised with micro-computed tomography

The skeleton morphology of the azooxanthellate cold-water coral Lophelia pertusa can be strongly influenced by invasive boring sponges that infest corallites in the still living part of the colony. Atypically swollen corallites of live Lophelia pertusa from the Galway Mound (Belgica Carbonate Mound Province, Porcupine Seabight, NE Atlantic), heavily excavated by boring organisms, have been examined with a wide range of non-destructive and destructive methods: micro-computed tomography, macro- and microscopic observations of the outer coral skeleton, longitudinal and transversal thin sections and SEM analyses of coral skeleton casts. As a result, three excavating sponge species have been distinguished within the coral skeleton: Alectona millari, Spiroxya heteroclita and Aka infesta. Furthermore, four main coral/sponge growth stages have been recognised: (1) cylindrical juvenile corallite/no sponge cavities; (2) flared juvenile corallite/linear sponge cavities (if present); (3) slightly swollen adult corallites/chambered oval sponge cavities; (4) very swollen adult corallites/widespread cavities. The inferred correlation between corallite morphology and boring sponge infestation has been detected in micro-computed tomography (micro-CT) images and confirmed in sponge trace casts and peculiar features of coral skeleton microstructure. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorised users.     

Cultivable Bacterial Community from South China Sea Sponge as Revealed by DGGE Fingerprinting and 16S rDNA Phylogenetic Analysis

The cultivable bacterial communities associated with four South China Sea sponges—Stelletta tenuis, Halichondria rugosa, Dysidea avara, and Craniella australiensis in mixed cultures—were investigated by microbial community DNA-based DGGE fingerprinting and 16S rDNA phylogenetic analysis. Diverse bacteria such as α-, γ-, δ-Proteobacteria, Bacteroidetes, and Firmicutes were cultured, some of which were previously uncultivable bacteria, potential novel strains with less than 95% similarity to their closest relatives and sponge symbionts growing only in the medium with the addition of sponge extract. According to 16S rDNA BLAST analysis, most of the bacteria were cultured from sponge for the first time, although similar phyla of bacteria have been previously recognized. The selective pressure of sponge extract on the cultured bacterial species was suggested, although the effect of sponge extract on bacterial community in high nutrient medium is not significant. Although α- and γ-Proteobacteria appeared to form the majority of the dominant cultivable bacterial communities of the four sponges, the composition of the cultivable bacterial community in the mixed culture was different, depending on the medium and sponge species. Greater bacterial diversity was observed in media C and CS for Stelletta tenuis, in media F and FS for Halichondria rugosa and Craniella australiensis. S. tenuis was found to have the highest cultivable bacterial diversity including α-, γ-, δ-Proteobacteria, Bacteroidetes, and Firmicutes, followed by sponge Dysidea avara without δ-Proteobacteria, sponge Halichondria rugosa with only α-, γ-Proteobacteria and Bacteroidetes, and sponge C. australiensis with only α-, γ-Proteobacteria and Firmicutes. Based on this study, by the strategy of mixed cultivation integrated with microbial community DNA-based DGGE fingerprinting and phylogenetic analysis, the cultivable bacterial community of sponge could be revealed effectively.     

Effects of freshwater sponge Ephydatia fluviatilis on conjugative transfer of antimicrobial resistance in Enterococcus faecalis strains in aquatic environments

Filter feeding is a biotic process that brings waterborne bacteria in close contact with each other and may thus support the horizontal transfer of their antimicrobial resistance genes. This laboratory study investigated whether the freshwater sponge Ephydatia fluviatilis supported the transfer of vancomycin resistance between two Enterococcus faecalis strains that we previously demonstrated to exhibit pheromone responsive plasmid conjugation. Microcosm experiments exposed live and dead colonies of laboratory - grown sponges to a vancomycin-resistant donor strain and a rifampicin-resistant recipient strain of Ent. faecalis. Enterococci with both resistance phenotypes were detected on double selection plates. In comparison to controls, abundance of these presumed transconjugants increased significantly in water from sponge microcosms. Homogenized suspensions of sponge cells also yielded presumed transconjugants; however, there was no significant difference between samples from live or dead sponges. Fluorescent in situ hybridization analysis of the sponge cell matrix using species-specific probes revealed the presence of enterococci clusters with cells adjacent to each other. The results demonstrated that sponge colonies can support the horizontal transfer of antimicrobial resistance although the mechanism underlying this process, such as binding of the bacteria to the sponge collagen matrix, has yet to be fully elucidated.     

Introductory comments

Abstract

Associated microorganisms have been described in numerous marine sponges. Their metabolic activity, however, has not yet been investigated in situ. We quantified for the first time microbial processes in a living sponge. Sulfate reduction rates of up to 1200 nmol cm^?3d^?1 were measured in the cold-water bacteriosponge Geodia barretti . Oxygen profiles and chemical analysis of sponge tissue and canal water revealed steep oxygen gradients and a rapid turnover of oxygen and sulfide, dependent on the pumping activity of the sponge. Identification of the microbial community with fluorescently labelled oligonucleotide probes (FISH) indicates the presence of sulfate-reducing bacteria belonging to the Desulfoarculus/Desulfomonile/Syntrophus -cluster in the choanosome of this sponge. Analysis of lipid biomarkers indicates biomass transfer from associated sulfate-reducing bacteria or other anaerobic microbes to sponge cells. These results show the presence of an anoxic micro-ecosystem in the sponge G. barretti, and imply mutualistic interactions between sponge cells and anaerobic microbes. Understanding the importance of anaerobic processes within the sponge/microbe system may help to answer unsolved questions in sponge ecology and biotechnology.     

Facies of Liassic sponge mounds, central High Atlas, Morocco

Summary Liassic sponge mounds of the central High Atlas (Rich area, northern Morocco) have a stratigraphic range from the Lower/Upper Sinemurian boundary interval up to the lower parts of the Lower Pliensbachian (Carixian). The base of Liassic sponge mounds consists of a transgressive discontinuity, i.e., a condensed section of microbioclastic wackestones with firm- and hardgrounds, ferruginous stromatolites, sponge spicules and ammonites. The top of Liassic sponge mounds is an irregular palaeorelief covered by cherty marl-limestone rhythmites, namely hemipelagic spicular wackestones with radiolaria. In the Rich area, section Foum Tillicht, the sponge mound succession has a total thickness of about 250 meters. Within this succession we distinguished between three mound intervals. The lower mound interval shows only small, meter-scale sponge mounds consisting of boundstones with lyssakine sponges, commensalicTerebella and the problematicumRadiomura. This interval forms a shallowing-upward sequence culminating in a bedded facies withTubiphytes, calcareous algae (Palaeodasycladus), sponge lithoclasts, coated grains, and thin rims of marine cement. The middle mound interval is aggradational with decametric mounds and distinct thrombolitic textures and reefal cavities. The mound assemblage here consists of hexactinellid sponges, lithistid demosponges, non-rigid demosponges,Radiomura, Serpula (Dorsoserpula), Terebella, encrusting bryozoa, and minor contributions by calcareous sponges, and excavating sponges (typeAka). Thrombolites are dendrolitic and may reach sizes of several tens of centimeters, similar to the maximum size of siliceous sponges. The upper mound interval appears retrogradational and geometries change upsection from mound shapes to flat lenses and level-bottom, biostromal sponge banks. The biotic assemblage is similar to that of the middle mound interval and there is no difference between mound and bank communities. The demise of sponge mounds is successive from regional spread in the Sinemurian to more localised spots in the Lower Pliensbachian. This reduction correlates with an increasing influence of pelagic conditions. At Foum Tillicht, sponge mounds lack any photic contribution and there is virtually no differentiation into subcommunities between mound surface and cavity dwelling organisms. There is some evidence that the heterotrophic food web of mound communities was sourced by oxygen minimum zone edge effects, namely microbial recycling of essential elements such as N and P. Basin geometry suggests a waterdepth of several 100's of meters, well below the photic zone and possibly only controlled by the depth range of the oxygen minimum zone. Palaeoceanographic conditions of well-stratified deeper water masses diminished gradually during widespread transgression across the Sinemurian to Pliensbachian boundary culminating in the Lower Pliensbachianibex ammonite zone.     

Trophodynamics of the sclerosponge Ceratoporella nicholsoni along a shallow to mesophotic depth gradient

The sclerosponge Ceratoporella nicholsoni is a hyper-calcifying high microbial abundance sponge. This sponge has been observed at high densities throughout the Caribbean in the mesophotic zone (30–150 m), as well as cryptic environments in shallow (< 30 m) depths. Given the densities of this sponge, it could play an important role in the cycling of inorganic and organic sources of carbon and nitrogen at mesophotic depths. Additionally, there is broad interest in this sponge as a tool for paleobiology, paleoclimatology and paleoceanography. As a result, it is increasingly important to understand the ecology of these unique sponges in the underexplored Caribbean mesophotic zone. Here we show that this sponge increases in abundance from shallow depths into the mesophotic zone of Grand Cayman Island. We observed no significant differences in the stable isotope signatures of δ¹⁵N and δ¹³C of sponge tissue between depths. A predictive model of sponge diet with increasing depth shows that these sponges consume dissolved organic matter of algal and coral origin, as well as the consumption of particulate organic matter consistent with the interpretation of the stable isotope data. The taxonomic composition of the sclerosponge microbiome was invariant across the shallow to mesophotic depth range but did contain the Phylum Chloroflexi, known to degrade a variety of dissolved organic carbon sources. These data suggest that the depth distribution of this sponge may not be driven by changes in trophic strategy and is potentially regulated by other biotic or abiotic factors.

     

Non-rigid cryptic sponges in oyster patch reefs (Lower Kimmeridgian, Langenberg/Oker, Germany)

Summary Two patch reefs which predominately consist of the oysterNanogyra nana (Sowerby 1822) are exposed in Lower Kimmeridigian strata of the Langenberg hillrange, central Germany. Left oyster valves making up the frame-work of the reefs formed small abundant cavities that were inhabited by a unique sponge community. The excellent preservation of non-rigid sponges was related to early organomineralization within the decaying sponge tissue. As a process of sponge taphonomy, different types of microbially induced carbonates precipitated preserving spicule aggregates. Organomineralization within sponge soft tissues is especially favored with the Langenberg patch reefs due to the closed or semi-closed system conditions with the cavities. The δ¹³C values ofin situ formed microbialities reveal that carbonate precipitation was in equilibrium with Jurassic seawater. The carbon of the microbialites does not derive from the bacterial remineralization of organic matter, but is of a marine source. Likewise, organomineralization is probably related to bacterial EPS or decaying sponge tissues providing an organic matrix for initial carbonate precipitation. Biomarker analyses revealed, that the patch reef microbialites contain terminally branched fatty acids (iso-andanteiso-pentadecanoic acid) in significant concentrations. These fatty acids, like hopanoid hydrocarbons, are most likely of a bacterial source. This is in agreement with sulfate-reducing bacteria remineralizing the decaying sponges as further indicated by the occurrence of framboidal pyrite in sponge microbialites.     

Culturable heterotrophic bacteria from the marine sponge <Emphasis Type="Italic">Dendrilla</Emphasis> <Emphasis Type="Italic">nigra</Emphasis>: isolation and phylogenetic diversity of actinobacteria

Culturable heterotrophic bacterial composition of marine sponge Dendrilla nigra was analysed using different enrichments. Five media compositions including without enrichment (control), enriched with sponge extract, with growth regulator (antibiotics), with autoinducers, and complete enrichment containing sponge extract, antibiotics, and autoinducers were developed. DNA hybridization assay was performed to explore host specific bacteria and ecotypes of culturable sponge-associated bacteria. Enrichment with selective inducers (AHLs and sponge extract) and regulators (antibiotics) considerably enhanced the cultivation potential of sponge-associated bacteria. It was found that Marinobacter (MSI032), Micromonospora (MSI033), Streptomyces (MSI051), and Pseudomonas (MSI057) were sponge-associated obligate symbionts. The present findings envisaged that “Micromonospora–Saccharomonospora–Streptomyces” group was the major culturable actinobacteria in the marine sponge D. nigra. The DNA hybridization assay was a reliable method for the analysis of culturable bacterial community in marine sponges. Based on the culturable community structure, the sponge-associated bacteria can be grouped (ecotypes) as general symbionts, specific symbionts, habitat flora, and antagonists.     

Microbial symbionts of the Australian Great Barrier Reef sponge, Candidaspongia flabellata

Microbial symbionts of the newly described rare, biochemically active Dictyoceratid sponge, Candidaspongia flabellata (Very White Fan) found in the Australian Great Barrier Reef, are being studied in detail. The chemistry of this sponge species is distinctive, and includes a previously undescribed compound, fanolide as well as homosesterterpene and bishomoscalarane secondary metabolites (Bergquist et al., 1999). Current research is focused on assessing the diversity of the microbial community associated with this sponge. The entire culturable community of this sponge has been studied in detail. A total of 228 bacteria, 25 fungi, 3 actinomycetes and 9 cyanobacteria were isolated from 10 individuals of this sponge. Eight eubacteria (designated AB001–AB008), along with seven cyanobacteria were consistently found associated with C. flabellata and absent from the surrounding water column, suggesting that these bacteria have a specific association with the sponge. Partial 16S ribosomal RNA gene sequencing of these isolates was done for phylogenetic characterisation. Electron microscopy was also used to confirm the presence of many morphotypes of bacteria and indicated spatial arrangements of particular morphotypes.     

L-Lactic acid production from raw cassava starch in a circulating loop bioreactor with cells immobilized in loofa (Luffa cylindrica)

l-Lactic acid was produced from raw cassava starch, by simultaneous enzyme production, starch saccharification and fermentation in a circulating loop bioreactor with Aspergillus awamori and Lactococcus lactis spp. lactis immobilized in loofa sponge. A. awamori was immobilized directly in cylindrical loofa sponge while the L. lactis was immobilized in a loofa sponge alginate gel cube. In the loofa sponge alginate gel cube, the sponge serves as skeletal support for the gel with the cells. The alginate gel formed a hard outer layer covering the soft porous gel inside. By controlling the rate and frequency of broth circulation between the riser and downcomer columns, the riser could be maintained under aerobic condition while the downcomer was under anaerobic condition. Repeated fed-batch l-lactic acid production was performed for more than 400 h and the average lactic acid yield and productivity from raw cassava starch were 0.76 g lactic acid g^–1 starch and 1.6 g lactic acid l^–1 h^–1, respectively.