Potential chemical defenses of Antarctic sponges against sympatric microorganisms

This study analyzed the bioactivity of extracts from 25 Antarctic demosponge species against 20 bacterial isolates and 1 diatom species collected from the waters off the western Antarctic Peninsula. All sponge species had lipophilic and hydrophilic extracts assayed at two concentrations (1× and 3× natural concentration) against 16 strains of Gamma Proteobacteria, 1 Flavobacterium, and 3 unidentified species of bacteria isolated from sympatric sponges. The majority of the bacterial isolates had no growth inhibition with only one isolate found to consistently have any growth inhibition due to sponge extracts. The sponges also had lipophilic and hydrophilic extracts assayed at three concentrations (0.3×, 1×, and 3× natural concentration) against the chain-forming pennate diatom Syndroposis sp. Almost every sponge’s lipophilic extract (96%) resulted in significant diatom mortality at the estimated natural concentration with the majority of the extracts (60%) still resulting in significant mortality at 30% of the natural concentration. The hydrophilic extracts of 60% of the sponges resulted in significant diatom mortality at the natural concentration. Even at 30% of the natural concentration, 24% of the hydrophilic sponge extracts resulted in significant diatom mortality. These sponges appear to have stronger defenses against diatom fouling than bacterial growth suggesting that there may be more selective pressure for chemical defenses against diatoms.     

Biofilm diatom community structure: Influence of temporal and substratum variability

Abstract

Diatoms, which are early autotrophic colonisers, are an important constituent of the biofouling community in the marine environment. The effects of substratum and temporal variations on the fouling diatom community structure in a monsoon-influenced tropical estuary were studied. Fibreglass and glass coupons were exposed every month for a period of 4 days and the diatom population sampled at 24 h intervals, over a period of 14 months. The planktonic diatom community structure differed from the biofilm community. Pennate diatoms dominated the biofilms whilst centric diatoms were dominant in the water column. Among the biofilm diatoms, species belonging to the genera Navicula, Amphora, Nitzschia, Pleurosigma and Thalassionema were dominant. On certain occasions, the influence of planktonic blooms was also seen on the biofilm community. A comparative study of biofilms formed on the two substrata revealed significant differences in density and diversity. However species composition was almost constant. In addition to substratum variations, the biofilm diatom community structure also showed significant seasonal variations, which were attributed to physico-chemical and biological changes in both the water and substratum. Temporal variations in the tychopelagic diatoms of the water were also observed to exert an influence on the biofilm diatom community. Variations in diatom communities may determine the functional ecosystem of the benthic environment.     

Multiple defensive roles for triterpene glycosides from two Caribbean sponges

Despite their high nutritional value and a lack of physical defenses, most marine sponges appear to be minimally affected by predators, competitors, and fouling organisms, possibly due to sponge chemical defenses. In the last 15 years, several triterpene glycosides have been isolated from sponges, but their ecological or physiological roles are largely unknown. We tested triterpene glycosides from Erylus formosus and Ectyoplasia ferox, Caribbean sponges belonging to two different orders, in field and laboratory assays for effects on fish feeding, attachment by potential biofilm-forming bacteria, fouling by invertebrates and algae, and overgrowth by neighboring sponges. Formoside and other triterpene glycosides from Erylus formosus deterred predation, microbial attachment, and fouling by invertebrates and algae. Triterpene glycosides from Ectyoplasia ferox were found to be antipredatory and allelopathic. Thus, triterpene glycosides in these sponges appear to have multiple ecological functions. Tests with different triterpene glycosides at several concentrations indicated that small differences in molecular structure affect ecological activity. In order to establish whether triterpene glycosides could be involved in water-borne versus surface-mediated interactions, the presence of triterpene glycosides in the seawater surrounding live sponges was measured using two in situ sampling methods followed by HPLC and NMR spectral analysis. Water-borne triterpene glycosides were below detection limits for both species. However, top sponge layers and swabs of the surfaces of both sponges contained sufficiently high concentrations of triterpene glycosides to deter bacterial settlement and fouling of Erylus formosus surfaces and overgrowth of Ectyoplasia ferox by neighboring sponges. Enemies of these sponges appear to be deterred by surface contact of triterpene glycosides rather than by water-borne interactions. The dual strategy of employing one group of compounds for multiple purposes and minimizing the loss of compounds into seawater suggests that these organisms utilize chemical defenses with efficiency.     

Marine antifoulants from bifurcaria bifurcata (phaeophyceae,cystoseiraceae) and other brown macroalgae

In this study, the antifouling activity of a series of extracts and linear diterpenes isolated from Bifurcaria bifurcata (Velley) Ross, a common brown alga of the Atlantic shores of Europe, and derivatives of these compounds was investigated. Antifouling assays with crude extracts from other brown algae, found abundantly along the coast of South Africa (Bifurcaria brassicaeformis, Bifurcariopsis capensis), the Atlantic shores of Europe (Halidrys siliquosa) and the coast of Mediterranean sea (Cladostephus verticillatus, Halopteris scoparia), are also reported. The fractions were tested in laboratory assays against representative species of the major groups of fouling organisms, viz. bacteria, fungi, diatoms, spores and zygotes of macroalgae and the blue mussel Mytilus edulis. Several components showed promising levels of activity. The high, albeit variable, level of antifouling activity suggests a potential for novel active ingredients in antifouling preparations.     

Antidiatom activity of marine bacteria associated with sponges from San Juan Island,Washington

Crude extracts of 52 marine bacteria associated with sponges, which were collected from the sea near San Juan Island, Washington, USA, were screened using diatom attachment assays against Amphora sp., Nitzschia closterium, Sellaphora sp. and Stauroneis sp. to investigate their antidiatom activities. Among these samples, five expressed strong anti-adhesion effects on all four tested diatoms. There was no negative effect observed from those five active samples on the growth of Amphora sp. Those five active samples were prepared from respective isolates, which all belonged to the genus Bacillus based on 16S rRNA gene sequencing analysis. The results of present study indicate that Bacillus may play important roles for sponges’ chemical defence against biofouling of diatoms and that the metabolites of Bacillus may be a potential source of natural antifouling compounds.     

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.     

Does concentrating chemical defenses within specific regions of marine sponges result in enhanced protection from predators?

Chemical defenses are an effective mode of predator deterrence across benthic marine organisms, but their production may come with associated costs to the organism as limited resources are diverted away from primary processes like growth and reproduction. Organisms concentrating ecologically relevant levels of these defenses in tissues most at risk to predator attack may alleviate this cost while deterring predators. We addressed this hypothesis by investigating the deterrence of chemical extracts from the inner and outer regions of the sponges Aplysina fulva, Ircinia felix, and I. campana from a temperate hard-bottom reef in the South Atlantic Bight. Assays were conducted using natural fish assemblages and sea urchins. Although, A. fulva and I. felix have higher concentrations of defensive metabolites in the outer and inner regions, respectively, extracts from these regions did not display enhanced deterrency against fish or mobile invertebrate predators. Likewise, extracts from both regions of the sponge Ircinia campana, which has a uniform distribution of defensive chemicals throughout, did not differ in their ability to deter either group of predators. Since chemical defenses were effective deterrents at lower concentrations, secondary metabolite allocation patterns observed among these sponges are likely not driven by predation pressure from generalist fish and mobile invertebrate predators on these reefs. Alternatively, these patterns may be driven by other ecological stressors, another suite of predators, or may be more effective at deterring predators when combined with structural defenses.     

Seasonal variations in the fouling diatom community structure from a monsoon influenced tropical estuary

Seasonal variations in the fouling diatom community from a monsoon influenced tropical estuary were investigated. The community composition did not differ significantly between stainless steel and polystyrene substrata due to dominance by Navicula spp. The experimental evidence suggests that Amphora, which is a dominant fouling diatom in temperate waters, ultimately dominates the community in tropical regions when conditions are favourable. These observations reveal that a faster onset of macrofouling interferes with the microfouling community wherein the faster recruiters that have a higher abundance in ambient waters, dominate the community. Seasonal variations were observed in the fouling diatom community. Navicula delicatula dominated during the post-monsoon and tychopelagic diatoms (Melosira and Odontella) were observed during the monsoon. Low diatom abundance was recorded during the pre-monsoon season. The results indicate that although the fouling diatom community composition does not vary between substrata, there is a seasonal change in the community depending on the physical, chemical and biological interactions.     

The diatoms ingested by freshwater snails: temporal,spatial, and interspecific variation

Seventeen species of diatoms, representing a broad range of sizes, shapes, and growth habits, were collected from rocks in rapidly-flowing sections of the Mitchell River, North Carolina. The diatoms ingested by adult Goniobasis proxima, juvenile Leptoxis carinata, and adult Physa sp. co-occurring in this habitat were indistinguishable from one another, in spite of great differences in radular morphology. All snails sampled the diatom flora almost randomly, with only one or two of the larger diatom species under-represented in the gut contents. Some snails also seemed to selectively ingest the smaller individuals of the larger diatom taxa, and larger individuals of the smaller diatom taxa. The diatoms identifiable in juvenile Goniobasis guts were somewhat more distinctive, although this seemed to be due at least partly to more mechanical breakage. The diatom flora of quiet, muddy pools was much different from that of shallow, rocky areas, but once again, Goniobasis seemed to sample the available flora randomly. Seasonal variation was also apparent in the diatom diet of Goniobasis. We suggest that in some cases, it may be reasonable to use snails to sample the diatom assemblage present in a particular habitat, if more direct methods are impractical.     

Testing attachment point theory: diatom attachment on microtextured polyimide biomimics

Abstract

This paper explores diatom attachment to a range of laser etched polyimide surfaces to directly test ‘attachment point theory’. Static bioassays were conducted on microtextured polyimide surfaces using four diatom species, Fallacia carpentariae, Nitzschia cf. paleacea, Amphora sp. and Navicula jeffreyi with cell sizes ranging from 1 – 14 μm. The microtextured polyimides were modelled from natural fouling resistant bivalve surfaces and had wavelengths above, below and at the same scale as the diatom cell sizes. Diatoms attached in significantly higher numbers to treatments where the numbers of attachment points was highest. The lowest diatom attachment occurred where cells were slightly larger than the microtexture wavelength, resulting in only two theoretical points of attachment. The results support attachment point theory and highlight the need to address larval/cell size in relation to the number of attachment points on a surface. Further studies examining a range of microtexture scales are needed to apply attachment point theory to a suite of fouling organisms and to develop structured surfaces to control the attachment and development of fouling communities.     

Mediterranean karst springs: diatom biodiversity hotspots under the pressure of hydrological fluctuation and nutrient enrichment

Abstract

Mediterranean karst springs are affected by strong climatic seasonality, with long, dry and hot summers and are increasingly threatened by anthropogenic pressures. In Sardinia (western Mediterranean Sea, Italy), they are largely unexplored and their biocoenoses are mostly unknown. The diatom flora from two substrates (cobbles and macrophytes) in eight springs of different areas of the island was investigated in summer 2016 and winter 2017. A total of 162 diatom taxa (58 genera) were found of which 27 (17 genera) only on cobbles and 26 (18 genera) only on macrophytes. The most abundant species from both substrates were Achnanthidium minutissimum, Amphora indistincta, Amphora pediculus, Cocconeis euglypta, Planothidium frequentissimum and P. lanceolatum. Overall, 67 taxa (40 genera) were recorded in single sites and some of these taxa showed high affinity with specific environmental conditions. Hydrological stability (water flow permanence), discharge and nutrients were the main environmental factors influencing diatom assemblages. Our results suggest that diatoms can reflect important local factors related to the vulnerability of these spring ecosystems and underline the importance of their preservation both for biodiversity and water quality maintenance.     

Succession and physiological health of freshwater microalgal fouling in a Tasmanian hydropower canal

Freshwater microalgal biofouling in hydropower canals in Tarraleah, Tasmania, is dominated by a single diatom species, Gomphonema tarraleahae. The microfouling community is under investigation with the aim of reducing its impact on electricity generation. Species succession was investigated using removable glass slides. Fouled slides were examined microscopically and for chlorophyll a biomass. Chl a biomass increased steeply after 8 weeks (0.09–0.87 mg m^?2), but increased much earlier on slides surrounded by a biofouled inoculum. Succession began with low profile diatoms such as Tabellaria flocculosa, progressing to stalked diatoms such as Gomphonema spp. and Cymbella aspera. Few chlorophytes and no filamentous algae were present. Pulse amplitude modulated fluorometry was used to measure the physiological health of fouling on the canal wall. Maximum quantum yield (F _v/F _m) measurements were consistently <0.18, indicating that the fouling mat consisted of dead or dying algae. The succession and physiological health of cells in the fouling community has broad implications for mitigation techniques used.     

Le diatomee bentoniche del porto di Palermo

Abstract

Benthic diatoms in the harbour of Palermo (Sicily).—The first results of a systematic investigation on the benthic diatoms on the fouling settlements, at four selected sites in the harbour of Palermo, are reported. Samples of comparable areas have been examined either on the fouling of vertical walls of the wharfs or on asbestos panels vertically submerged at the same place for at least eighteen months. Seventy-seven species have been found. No marked taxonomic difference is evident between the populations on the stoneworks and those on the panels.     

A simple method to evaluate the potential for degradation of antifouling biocides

A simple method to measure the degradation of antifouling biocides is described which measures the loss of biocidal activity from seawater by bioassay. The bioassay employs either the ship‐fouling diatom Amphora or the brine shrimp Anemia. Loss of bioaclivity from sterile seawater indicates abiotic degradation whilst loss of bioactivity from natural seawater indicates biodegradation. Results are presented for three biocides, viz. the trihalomethylthio compound, N‐dichlorofluoromethylthio‐N’,N'‐dimethyl‐N‐phenyl‐sulphamide (Preventol A4S), di‐n‐octylamine, and the isothiazolone compound 4,5‐dichloro‐2‐n‐octyl‐4‐isothiazolin‐3‐one (Sea‐Nine 211).     

Adhesion and motility of fouling diatoms on a silicone elastomer

Recent demands for non-toxic antifouling technologies have led to increased interest in coatings based on silicone elastomers that ‘release’ macrofouling organisms when hydrodynamic conditions are sufficiently robust. However, these types of coatings accumulate diatom slimes, which are not released even from vessels operating at high speeds ( > 30 knots). In this study, adhesion strength and motility of three common fouling diatoms (Amphora coffeaeformis var. perpusilla (Grunow) Cleve, Craspedostauros australis Cox and Navicula perminuta Grunow) were measured on a polydimethylsiloxane elastomer (PDMSE) and acid-washed glass. Adhesion of the three species was stronger to PDMSE than to glass but the adhesion strengths varied. The wall shear stress required to remove 50% of cells from PDMSE was 17 Pa for Craspedostauros, 24 Pa for Amphora and >> 53 Pa for Navicula; the corresponding values for glass were 3, 10 and 25 Pa. In contrast, the motility of the three species showed little or no correlation between the two surfaces. Craspedostauros moved equally well on glass and PDMSE, Amphora moved more on glass initially before movement ceased and Navicula moved more on PDMSE before movement ceased. The results show that fouling diatoms adhere more strongly to a hydrophobic PDMSE surface, and this feature may contribute to their successful colonization of low surface energy, foul-release coatings. The results also indicate that diatom motility is not related to adhesion strength, and motility does not appear to be a useful indicator of surface preference by diatoms.     

THE ROLE OF THE REACTIVITY AND CONTENT OF IRON OF AEROSOL DUST ON GROWTH RATES OF TWO ANTARCTIC DIATOM SPECIES1

The atmosphere is widely recognized as a major source of Fe in the form of iron‐containing dust. This study provides the first experiments in which the impact of dust on the growth rates of single species of Antarctic diatoms was assessed under laboratory conditions. The dust was among others characterized by x‐ray powder diffraction analysis, analysis of total and amorphous Fe content, and dissolution rates of Fe in seawater. The amount of bioavailable Fe from the dust was determined, not via the complicated chemistry of Fe in seawater but by using diatoms as bioindicators for available Fe. Cultures of two large diatom species, Actinocyclus sp. and Thalassiosira sp., were amended with potential dust aerosols from two dust‐supplying regions, Namibia and Mauritania, and responses on growth rates were monitored. Apart from a difference in total Fe content, a difference in crystallinity existed in the Fe minerals of both dust types. The fraction of amorphous Fe was reflected in a higher reactivity/dissolution of Fe in seawater. The increase in growth rate upon dust addition was positively related with the amount of amorphous Fe in the dust and with the dissolution rate of Fe in seawater. However, compared with equal FeCl₃ concentrations, the dissolved Fe from the dust was not completely available for the diatoms. Interestingly, the diatoms used only a small part of the dissolved Fe, demonstrating the importance of algae as bioindicators.     

Static vs dynamic settlement and adhesion of diatoms to ship hull coatings

Many experiments utilize static immersion tests to evaluate the performance of ship hull coatings. These provide valuable data; however, they do not accurately represent the conditions both the hull and fouling organisms encounter while a ship is underway. This study investigated the effect of static and dynamic immersion on the adhesion and settlement of diatoms to one antifouling coating (BRA 640), four fouling-release coatings (Intersleek^® 700, Intersleek^® 900, Hempasil X3, and Dow Corning 3140) and one standard surface (Intergard^® 240 Epoxy). Differences in community composition were observed between the static and dynamic treatments. Achnanthes longipes was present on all coatings under static immersion, but was not present under dynamic immersion. This was also found for diatoms in the genera Bacillaria and Gyrosigma. Melosira moniformis was the only diatom present under dynamic conditions, but not static conditions. Several common fouling diatom genera were present on panels regardless of treatment: Amphora, Cocconeis, Entomoneis Cylindrotheca, Licmophora, Navicula, Nitzschia, Plagiotropis, and Synedra. Biofilm adhesion, diatom abundance and diatom diversity were found to be significantly different between static and dynamic treatments; however, the difference was dependent on coating and sampling date. Several coatings (Epoxy, DC 3140 and IS 700) had significantly higher biofilm adhesion on dynamically treated panels on at least one of the four sampling dates, while all coatings had significantly higher diatom abundance on at least one sampling date. Diversity was significantly greater on static panels than dynamic panels for Epoxy, IS 700 and HX3 at least once during the sampling period. The results demonstrate how hydrodynamic stress will significantly influence the microfouling community. Dynamic immersion testing is required to fully understand how antifouling surfaces will respond to biofilm formation when subjected to the stresses experienced by a ship underway.     

Antidiatom and antibacterial activity of epiphytic bacteria isolated from <Emphasis Type="Italic">Ulva</Emphasis><Emphasis Type="Italic">lactuca</Emphasis> in tropical waters

Bacteria and diatoms are primary colonizers of marine surfaces and hence play a crucial role in the attachment and subsequent growth of macroorganisms. It has been suggested that the temperate green alga Ulva lactuca relies on the defence provided by the epiphytic bacterial community to regulate surface fouling of colonising organisms. In this study, ten resident bacterial isolates from tropical U. lactuca were tested for their antibacterial and antidiatom properties that may regulate surface colonization on the algae. Sixty percent of the epiphytic isolates expressed antibacterial properties against other resident bacteria and 80% had antidiatom activity against the pennate diatom, Cylindrotheca fusiformis. Isolates of the Pseudoalteromonas genus showed both- antibacterial and antidiatom activities, while members of the genus Bacillus, Vibrio and Shewanella mostly possessed antidiatom activity. Our results show that a high proportion of bacterial isolates from tropical U. lactuca, like that of their temperate counterparts contain antibiotic properties that might impact on the bacterial community composition and prevent fouling by diatoms.     

Growth limitation of three Arctic sea ice algal species: effects of salinity, pH, and inorganic carbon availability

The effect of salinity, pH, and dissolved inorganic carbon (TCO₂) on growth and survival of three Arctic sea ice algal species, two diatoms (Fragilariopsis nana and Fragilariopsis sp.), and one species of chlorophyte (Chlamydomonas sp.) was assessed in controlled laboratory experiments. Our results suggest that the chlorophyte and the two diatoms have different tolerance to fluctuations in salinity and pH. The two species of diatoms exhibited maximum growth rates at a salinity of 33, and growth rates at a salinity of 100 were reduced by 50% compared to at a salinity of 33. Growth ceased at a salinity of 150. The chlorophyte species was more sensitive to high salinities than the two diatom species. Growth rate of the chlorophyte was greatly reduced already at a salinity of 50 and it could not grow at salinities above 100. At salinity 33 and constant TCO₂ concentration, all species exhibited maximal growth rate at pH 8.0 and/or 8.5. The two diatom species stopped growing at pH > 9.5, while the chlorophyte species still was able to grow at a rate which was 1/3 of its maximum growth rate at pH 10. Thus, Chlamydomonas sp. was able to grow at high pH levels in the succession experiment and therefore outcompeted the two diatom species. Complementary experiments indicated that growth was mainly limited by pH, while inorganic carbon limitation only played an important role at very high pH levels and low TCO₂ concentrations.     

Testing for defensive synergy in Caribbean sponges: Bad taste or glass spicules?

Chemical and physical defenses of sessile organisms against consumers are well described for both terrestrial and marine systems. However, previous studies have focused on chemical or physical defenses in isolation, and have not considered their interaction. Marine sponges provide a model system for testing this interaction. Some sponge species produce secondary metabolites that deter predation; they may also contain siliceous spicules, but previous studies have provided little evidence that spicules in isolation offer any defense against generalist fish predators. To determine whether the two components have an additive, antagonistic, or synergistic interaction, crude organic extracts and spicules from individuals of 8 Caribbean sponge species were isolated and tested in laboratory feeding assays. These included one chemically defended reef sponge (Agelas clathrodes) and seven known to be intermediately deterrent: six from reef habitats (Cinachyrella alloclada, Clathria virgultosa, Cribrochalina infundibulum, Niphates digitalis, Svenzea zeai, and Xestospongia muta) and one from mangrove habitats (Tedania ignis). Extracts and spicules were assayed at various concentrations, both individually and in combination, in laboratory feeding assays with the bluehead wrasse, Thalassoma bifasciatum. A SAS based GENMOD procedure based on an isobolographic analysis model was used for statistical comparisons. Four sponges (A. clathrodes, C. alloclada, C. virgultosa, and one of three individuals of X. muta) showed evidence of synergisms. Of these, synergy in C. alloclada, C. virgultosa, and X. muta was caused by approximately natural concentrations of extracts and spicules. The extract of A. clathrodes was deterrent, but combination assays required nearly a 3-fold reduction in extract concentration and an 8-fold increase in spicule concentration to show the synergistic effect. Contrary to previous findings, spicules from C. infundibulum and two of three individuals of X. muta were deterrent at natural concentrations. Sponge spicules may be defensive in isolation, or may enhance chemical defenses against consumers, but the lack of synergisms for individuals in 4 of 7 species with intermediate levels of chemical defense suggests that defensive synergy is not the general rule and, when present, may be an example of an exaptation.