Engineered living photosynthetic biocomposites for intensified biological carbon capture.

Carbon capture and storage is required to meet Paris Agreement targets. Photosynthesis is nature's carbon capture technology. Drawing inspiration from lichen, we engineered 3D photosynthetic cyanobacterial biocomposites (i.e., lichen mimics) using acrylic latex polymers applied to loofah sponge. Biocomposites had CO2 uptake rates of 1.57 ± 0.08 g CO2 g-1biomass d-1. Uptake rates were based on the dry biomass at the start of the trial and incorporate the CO2 used to grow new biomass as well as that contained in storage compounds such as carbohydrates. These uptake rates represent 14-20-fold improvements over suspension controls, potentially scaling to capture 570 tCO2 t-1biomass yr-1, with an equivalent land consumption of 5.5-8.17 × 106 ha, delivering annualized CO2 removal of 8-12 GtCO2, compared with 0.4-1.2 × 109 ha for forestry-based bioenergy with carbon capture and storage. The biocomposites remained functional for 12 weeks without additional nutrient or water supplementation, whereupon experiments were terminated. Engineered and optimized cyanobacteria biocomposites have potential for sustainable scalable deployment as part of humanity's multifaceted technological stand against climate change, offering enhanced CO2 removal with low water, nutrient, and land use penalties.

Lesser weever fish (Echiichthys vipera Cuvier, 1829) venom is cardiotoxic but not haemorrhagic.

Despite comprising over half of the biodiversity of living venomous vertebrates, fish venoms are comparatively understudied. Venom from the lesser weever fish (Echiichthys vipera syn. Trachinus vipera) has received only cursory attention despite containing one of the most potent venom toxins (trachinine). Literature records are further complicated by early studies combining the venom with that of the related greater weever (Trachinus draco). The current study used a chicken chorioallantoic membrane assay to investigate venom bioactivity following the application of measured quantities of crude venom to a major bilateral vein at 1 cm distance from the heart. The venom had a dose-dependent effect on survival rate and exhibited dose-dependent cardiotoxic properties at day six of development. Crude E. vipera triggered tachycardia at doses of 37.58 and 44.88 µg/µL and bradycardia at 77.4 µg/µL. The three highest doses (65.73, 77.4 and 151.24 µg/µL) caused significant mortality. These data also suggested intra-specific variation in E. vipera venom potency. Unlike a number of other piscine venoms, E. vipera venom was not haemorrhagic at the concentrations assayed.

The venoms of the lesser (Echiichthys vipera) and greater (Trachinus draco) weever fish- A review.

In comparison with other animal venoms, fish venoms remain relatively understudied. This is especially true for that of the lesser Echiichthys vipera and greater weever fish Trachinus draco which, apart from the isolation of their unique venom cytolysins, trachinine and dracotoxin, respectively, remain relatively uncharacterised. Envenomation reports mainly include mild symptoms consisting of nociception and inflammation. However, like most fish venoms, if the venom becomes systemic it causes cardiorespiratory and blood pressure changes. Although T. draco venom has not been studied since the 1990's, recent studies on E. vipera venom have discovered novel cytotoxic components on human cancer cells, but due to the scarcity of research on the molecular make-up of the venom, the molecule(s) causing this cytotoxicity remains unknown. This review analyses past studies on E. vipera and T. draco venom, the methods used in the , the venom constituents characterised, the reported symptoms of envenomation and compares these findings with those from other venomous Scorpaeniformes.

Diatom-Derived Polyunsaturated Aldehydes Are Unlikely to Influence the Microbiota Composition of Laboratory-Cultured Diatoms.

Diatom-derived oxylipins, including polyunsaturated aldehydes (PUA), are considered to have infochemical, allelochemical and bacteriostatic properties, with plausible roles as grazing deterrents and regulators of inter- and intraspecific competition. However, the extent and mechanisms of how PUA influence diatom-bacteria interactions remain unresolved. In this study, impacts on the diversity of the associated bacterial communities (microbiota) of two contrasting Skeletonema marinoi strains (a PUA and a non-PUA producer) were investigated under three nitrate conditions in batch culture. Further, the response of the culture microbiota was studied when spiked with PUA at ecologically relevant concentrations (86nM octadienal and 290nM heptadienal). Of the 741 identified OTUs, Proteobacteria was the most abundant phylum (62.10%), followed by Bacteroidetes (12.33%) and Firmicutes (6.11%). Escherichia/Shigella were the most abundant genera for all treatments. Similar communities were present in both spiked and non-spiked cultures suggesting they can tolerate PUA exposure at realistic concentrations. This study suggests that PUA are not major drivers of diatom-bacteria interactions in laboratory cultures.

Co-digestion of microalgae with potato processing waste and glycerol: effect of glycerol addition on methane production and the microbial community.

The production of methane-rich biogas from the anaerobic digestion (AD) of microalgae is limited by an unfavorable biomass carbon-to-nitrogen (C/N) ratio; however, this may be ameliorated using a co-digestion strategy with carbon-rich feedstocks. For reliable plant operation, and to improve the economics of the process, secure co-feedstock supply (ideally as a waste-stream) is important. To this end, this study investigated the feasibility of co-digesting microalgae (Chlorella vulgaris) with potato processing waste (potato discarded parts, PPWdp; potato peel, PPWp) and glycerol, while monitoring the response of the methanogenic community. In this semi-continuous study, glycerol (1 and 2% v/v) added to mixtures of C. vulgaris : PPWdp enhanced the specific methane yields the most, by 53-128%, whilst co-digestion with mixtures of C. vulgaris : PPWp enhanced the methane yields by 62-74%. The microbial communities diverged markedly over operational time, and to a lesser extent in response to glycerol addition. The acetoclast Methanosaeta was abundant in all treatments but was replaced by Methanosarcina in the potato peel with glycerol treatment due to volatile fatty acid (VFA) accumulation. Our findings demonstrate that the performance of microalgae co-digestion is substantially improved by the addition of glycerol as an additional co-feedstock. This should improve the economic case for anaerobically digesting microalgae as part of wastewater treatment processes and/or the terminal step of a microalgae biorefinery.

Equistatin and equinatoxin gene expression is influenced by environmental temperature in the sea anemone Actinia equina.

We examined the gene expression levels of equinatoxin and equistatin in the sea anemone Actinia equina, when reared at varying environmental temperatures for five months. Both genes were significantly downregulated at 10 °C compared to 16 °C but showed no significant change at 22 °C. This provides the first evidence of an effect of temperature on gene expression, but with no effect of increasing temperatures such as those predicted due to climate change.

Anthropogenic noise compromises the anti-predator behaviour of the European seabass, Dicentrarchus labrax (L.).

Anthropogenic noise is a significant pollutant of the world's oceans, affecting behavioural and physiological traits in a range of species, including anti-predator behaviours. Using the open field test, we investigated the effects of recordings of piling and drilling noise on the anti-predator behaviour of captive juvenile European seabass in response to a visual stimulus (a predatory mimic). The impulsive nature of piling noise triggered a reflexive startle response, which contrasted the behaviour elicited by the continuous drilling noise. When presented with the predatory mimic, fish exposed to both piling and drilling noise explored the experimental arena more extensively than control fish exposed to ambient noise. Fish under drilling and piling conditions also exhibited reduced predator inspection behaviour. Piling and drilling noise induced stress as measured by ventilation rate. This study provides further evidence that the behaviour and physiology of European seabass is significantly affected by exposure to elevated noise levels.

Assessing the impact of diclofenac, ibuprofen and sildenafil citrate (Viagra®) on the fertilisation biology of broadcast spawning marine invertebrates.

Exposure to synthetic chemicals is a key environmental challenge faced by aquatic organisms. The time and dose effects of the pharmaceuticals diclofenac, ibuprofen, and sildenafil citrate on sperm motility and successful fertilisation are studied using the echinoderms, Asterias rubens and Psammechinus miliaris, and the polychaete worm Arenicola marina, all important components of the marine benthos. Motility was reduced for all species when exposed to diclofenac concentrations ≥0.1 µg/L. Exposure to ≥1.0 µg/L of ibuprofen affected only P. miliaris gametes and fertilisation success of A. marina. A. rubens and P. miliaris sperm increased in both percentage motility and swimming velocity when exposed to sildenafil citrate at concentrations ≥18 and ≥ 50 ng/L, respectively. Pre-incubation of sperm with sildenafil citrate significantly increased fertilisation success in A. rubens and P. miliaris but not in A. marina. Pre-incubated A. rubens oocytes fertilised successfully in ibuprofen. According to EU Directive 93/67/EEC, diclofenac is classified as a very toxic substance to gametes of A. rubens, P. miliaris, and A. marina (EC50 = 100-1000 µg/L) while ibuprofen is classified as very toxic to gametes of P. miliaris but non-toxic to gametes of A. marina (EC50 > 10,000 µg/L). The present study indicates that diclofenac exposure may have negative impacts on invertebrate reproductive success, whereas ibuprofen potentially may compromise P. miliaris reproduction. This study provides a valuable insight into the mechanisms that allow marine invertebrates to survive and reproduce in contaminated and changing habitats.

Eutrophication and warming-driven green tides (Ulva rigida) are predicted to increase under future climate change scenarios.

The incidence and severity of extraordinary macroalgae blooms (green tides) are increasing. Here, climate change (ocean warming and acidification) impacts on life history and biochemical responses of a causative green tide species, Ulva rigida, were investigated under combinations of pH (7.95, 7.55, corresponding to lower and higher pCO2), temperature (14, 18°C) and nitrate availability (6 and 150µmolL-1). The higher temperature accelerated the onset and magnitude of gamete settlement. Any two factor combination promoted germination and accelerated growth in young plants. The higher temperature increased reproduction, which increased further in combination with elevated pCO2 or nitrate. Reproductive success was highest (64.4±5.1%) when the upper limits of all three variables were combined. Biochemically, more protein and lipid but less carbohydrate were synthesized under higher temperature and nitrate conditions. These results suggest that climate change may cause more severe green tides, particularly when eutrophication cannot be effectively controlled.

Profiling bacterial communities associated with sediment-based aquaculture bioremediation systems under contrasting redox regimes.

Deposit-feeding invertebrates are proposed bioremediators in microbial-driven sediment-based aquaculture effluent treatment systems. We elucidate the role of the sediment reduction-oxidation (redox) regime in structuring benthic bacterial communities, having direct implications for bioremediation potential and deposit-feeder nutrition. The sea cucumber Holothuria scabra was cultured on sediments under contrasting redox regimes; fully oxygenated (oxic) and redox stratified (oxic-anoxic). Taxonomically, metabolically and functionally distinct bacterial communities developed between the redox treatments with the oxic treatment supporting the greater diversity; redox regime and dissolved oxygen levels were the main environmental drivers. Oxic sediments were colonised by nitrifying bacteria with the potential to remediate nitrogenous wastes. Percolation of oxygenated water prevented the proliferation of anaerobic sulphate-reducing bacteria, which were prevalent in the oxic-anoxic sediments. At the predictive functional level, bacteria within the oxic treatment were enriched with genes associated with xenobiotics metabolism. Oxic sediments showed the greater bioremediation potential; however, the oxic-anoxic sediments supported a greater sea cucumber biomass. Overall, the results indicate that bacterial communities present in fully oxic sediments may enhance the metabolic capacity and bioremediation potential of deposit-feeder microbial systems. This study highlights the benefits of incorporating deposit-feeding invertebrates into effluent treatment systems, particularly when the sediment is oxygenated.

MALDI-TOF Mass Spectrometry Discriminates Known Species and Marine Environmental Isolates of Pseudoalteromonas.

The genus Pseudoalteromonas constitutes an ecologically significant group of marine Gammaproteobacteria with potential biotechnological value as producers of bioactive compounds and of enzymes. Understanding their roles in the environment and bioprospecting for novel products depend on efficient ways of identifying environmental isolates. Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) biotyping has promise as a rapid and reliable method of identifying and distinguishing between different types of bacteria, but has had relatively limited application to marine bacteria and has not been applied systematically to Pseudoalteromonas. Therefore, we constructed a MALDI-TOF MS database of 31 known Pseudoalteromonas species, to which new isolates can be compared by MALDI-TOF biotyping. The ability of MALDI-TOF MS to distinguish between species was scrutinized by comparison with 16S rRNA gene sequencing. The patterns of similarity given by the two approaches were broadly but not completely consistent. In general, the resolution of MALDI-TOF MS was greater than that of 16S rRNA gene sequencing. The database was tested with 13 environmental Pseudoalteromonas isolates from UK waters. All of the test strains could be identified to genus level by MALDI-TOF MS biotyping, but most could not be definitely identified to species level. We conclude that several of these isolates, and possibly most, represent new species. Thus, further taxonomic investigation of Pseudoalteromonas is needed before MALDI-TOF MS biotyping can be used reliably for species identification. It is, however, a powerful tool for characterizing and distinguishing among environmental isolates and can make an important contribution to taxonomic studies.

Proteomic-based biotyping reveals hidden diversity within a microalgae culture collection: An example using Dunaliella.

Accurate and defendable taxonomic identification of microalgae strains is vital for culture collections, industry and academia; particularly when addressing issues of intellectual property. We demonstrate the remarkable effectiveness of Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry (MALDI-TOF-MS) biotyping to deliver rapid and accurate strain separation, even in situations where standard molecular tools prove ineffective. Highly distinctive MALDI spectra were obtained for thirty two biotechnologically interesting Dunaliella strains plus strains of Arthrospira, Chlorella, Isochrysis, Tetraselmis and a range of culturable co-occurring bacteria. Spectra were directly compared with genomic DNA sequences (internal transcribed spacer, ITS). Within individual Dunaliella isolates MALDI discriminated between strains with identical ITS sequences, thereby emphasising and enhancing knowledge of the diversity within microalgae culture collections. Further, MALDI spectra did not vary with culture age or growth stage during the course of the experiment; therefore MALDI presents stable and accurate strain-specific signature spectra. Bacterial contamination did not affect MALDI's discriminating power. Biotyping by MALDI-TOF-MS will prove effective in situations wherein precise strain identification is vital, for example in cases involving intellectual property disputes and in monitoring and safeguarding biosecurity. MALDI should be accepted as a biotyping tool to complement and enhance standard molecular taxonomy for microalgae.

The effect of bubble size on the efficiency and economics of harvesting microalgae by foam flotation.

The effect of bubble size and rise velocity on the efficiency of a foam flotation microalgae harvesting unit was determined. Three sparger and input airflow combinations were used: (1) limewood sparger with constant airflow, (2) ceramic flat plate sparger with constant airflow and (3) ceramic flat plate sparger with an oscillating airflow. The ceramic sparger with oscillating flow generated the smallest bubbles within the liquid pool and the largest bubbles within the foam phase. This delivered the highest levels of biomass recovery due to enhanced bubble-algae collision and attachment efficiencies. The smaller bubbles generated by the ceramic sparger under constant or oscillating airflow had significantly faster rise velocities when compared to the larger bubbles produced by the limewood spargers. The faster velocities of the smaller bubbles were due to momentum transfer to the liquid phase. Analyses of the harvest economics revealed that the ceramic flat plate sparger with an oscillating airflow delivered the best overall cost-benefit relationship.

Response of copepods to elevated pCO2 and environmental copper as co-stressors--a multigenerational study.

We examined the impacts of ocean acidification and copper as co-stressors on the reproduction and population level responses of the benthic copepod Tisbe battagliai across two generations. Naupliar production, growth, and cuticle elemental composition were determined for four pH values: 8.06 (control); 7.95; 7.82; 7.67, with copper addition to concentrations equivalent to those in benthic pore waters. An additive synergistic effect was observed; the decline in naupliar production was greater with added copper at decreasing pH than for decreasing pH alone. Naupliar production modelled for the two generations revealed a negative synergistic impact between ocean acidification and environmentally relevant copper concentrations. Conversely, copper addition enhanced copepod growth, with larger copepods produced at each pH compared to the impact of pH alone. Copepod digests revealed significantly reduced cuticle concentrations of sulphur, phosphorus and calcium under decreasing pH; further, copper uptake increased to toxic levels that lead to reduced naupliar production. These data suggest that ocean acidification will enhance copper bioavailability, resulting in larger, but less fecund individuals that may have an overall detrimental outcome for copepod populations.

Laboratory culture and evaluation of the tubeworm Ficopomatus enigmaticus for biofouling studies.

Ficopomatus enigmaticus, a euryhaline tube-building polychaete worm with a subtropical to temperate distribution, is an increasingly problematic fouling organism. In this study, laboratory protocols for maintaining adult broodstock, destructive spawning, larval culture and a settlement bioassay were developed. The method routinely yielded approximately 200 larvae per spawning adult. The mean number of eggs released by females was 1517 and the mean number of spermatozoids per male was 4.425 × 10(6). Fertilisation success, using an initial concentration of 2.5 × 10(6) spermatozoids and 45 eggs ml(-1), was 76% after a contact time of 60 min. The first cleavage occurred after 20 min and the trocophore larval stage was attained by 18 h. Metatrochophores were observed 4 d post-fertilisation and were competent to settle 1 day later. The proportion of larvae that settled after 48 h was surface-dependent: 10.24% on glass, 1.39% on polystyrene and 11.07% on a poly(dimethylsiloxane) elastomer. The presence of a biofilm on glass increased the rate of settlement 7-fold compared to clean glass.

The steady state anaerobic digestion of Laminaria hyperborea--effect of hydraulic residence on biogas production and bacterial community composition.

Methane production by anaerobic digestion (AD) of macroalgae (seaweed) is a promising algal bioenergy option. Work presented here is primarily based on the AD of Laminaria hyperborea using batch and continuously stirred tank reactors. Extrapolation of data from batch studies to long term continuous reactors was unreliable. A conservative organic loading rate (OLR) of 1 g L(-1) d(-1) was used due to difficulties experienced in achieving steady state performance at an OLR of 1.5 g L(-1) d(-1). Biogas composition and methane yields (60-70%) were near to values expected from terrestrial feedstocks. Biomass washout, as imposed by the dilution rate (i.e., hydraulic residence), had considerable bearing on the biogas generation profile, particularly at >3 hydraulic residences. Inhibition of methanogen growth was linked to nutrient deficiency and potentially antimicrobial compounds associated with the feedstock. Anaerobic digestion of L. hyperborea proved feasible over extended operational periods.

Treatment with algae extracts promotes flocculation, and enhances growth and neutral lipid content in Nannochloropsis oculata--a candidate for biofuel production.

Marine microalgae represent a potentially valuable feedstock for biofuel production; however, large-scale production is not yet economically viable. Optimisation of productivity and lipid yields is required and the cost of biomass harvesting and dewatering must be significantly reduced. Microalgae produce a wide variety of biologically active metabolites, many of which are involved in inter- and intra-specific interactions (the so-called infochemicals). The majority of infochemicals remain unidentified or uncharacterised. Here, we apply known and candidate (undefined extracts) infochemicals as a potential means to manipulate the growth and lipid content of Nannochloropsis oculata-a prospective species for biofuel production. Five known infochemicals (ß-cyclocitral, trans,trans-2,4-decadienal, hydrogen peroxide, norharman and tryptamine) and crude extracts prepared from Skeletonema marinoi and Dunaliella salina cultures at different growth stages were assayed for impacts on N. oculata over 24 h. The neutral lipid content of N. oculata increased significantly with exposure to three infochemicals (ß-cyclocitral, decadienal and norharman); however the effective concentrations affected a significant decrease in growth. Exposure to particular crude extracts significantly increased both growth and neutral lipid levels. In addition, water-soluble extracts of senescent S. marinoi cultures induced a degree of flocculation in the N. oculata. These preliminary results indicate that artificial manipulation of N. oculata cultures by application of algae infochemicals could provide a valuable tool towards achieving economically viable large-scale algae biofuel production.

Structural characterisation of the N-glycan moiety of the barnacle settlement-inducing protein complex (SIPC).

Many barnacle species are gregarious and their cypris larvae display a remarkable ability to explore surfaces before committing to permanent attachment. The chemical cue to gregarious settlement behaviour - the settlement-inducing protein complex (SIPC) - is an α(2)-macroglobulin-like glycoprotein. This cuticular protein may also be involved in cyprid reversible adhesion if its presence is confirmed in footprints of adhesive deposited during exploratory behaviour, which increase the attractiveness of surfaces and signal other cyprids to settle. The full-length open-reading frame of the SIPC gene encodes a protein of 1547 amino acids with seven potential N-glycosylation sites. In this study on Balanus amphitrite, glycan profiling of the SIPC via hydrophilic interaction liquid chromatography with fluorescence detection (HILIC-fluorescence) provided evidence of predominantly high mannose glycans (M2-9), with the occurrence of monofucosylated oligomannose glycans (F(6)M2-4) in lower proportions. The high mannose glycosylation found supports previous observations of an interaction with mannose-binding lectins and exogenous mannose increasing settlement in B. amphitrite cypris larvae. Transmission electron microscopy of the deglycosylated SIPC revealed a multi-lobed globular protein with a diameter of ~8 nm. Obtaining a complete structural characterisation of the SIPC remains a goal that has the potential to inspire solutions to the age-old problem of barnacle fouling.

The relevance of marine chemical ecology to plankton and ecosystem function: an emerging field.

Marine chemical ecology comprises the study of the production and interaction of bioactive molecules affecting organism behavior and function. Here we focus on bioactive compounds and interactions associated with phytoplankton, particularly bloom-forming diatoms, prymnesiophytes and dinoflagellates. Planktonic bioactive metabolites are structurally and functionally diverse and some may have multiple simultaneous functions including roles in chemical defense (antipredator, allelopathic and antibacterial compounds), and/or cell-to-cell signaling (e.g., polyunsaturated aldehydes (PUAs) of diatoms). Among inducible chemical defenses in response to grazing, there is high species-specific variability in the effects on grazers, ranging from severe physical incapacitation and/or death to no apparent physiological response, depending on predator susceptibility and detoxification capability. Most bioactive compounds are present in very low concentrations, in both the producing organism and the surrounding aqueous medium. Furthermore, bioactivity may be subject to synergistic interactions with other natural and anthropogenic environmental toxicants. Most, if not all phycotoxins are classic secondary metabolites, but many other bioactive metabolites are simple molecules derived from primary metabolism (e.g., PUAs in diatoms, dimethylsulfoniopropionate (DMSP) in prymnesiophytes). Producing cells do not seem to suffer physiological impact due to their synthesis. Functional genome sequence data and gene expression analysis will provide insights into regulatory and metabolic pathways in producer organisms, as well as identification of mechanisms of action in target organisms. Understanding chemical ecological responses to environmental triggers and chemically-mediated species interactions will help define crucial chemical and molecular processes that help maintain biodiversity and ecosystem functionality.

Exposure to copper and a cytotoxic polyunsaturated aldehyde induces reproductive failure in the marine polychaete Nereis virens (Sars).

A number of metabolites from microalgae, including polyunsaturated aldehydes (PUAs), have been implicated as inducers of reproductive failure in aquatic invertebrates. Current work describes the impacts of the model PUA 2E, 4E-decadienal and copper sulphate applied in isolation and combination on the reproductive performance of the infaunal polychaete, Nereis virens (Sars). The reproductive and life cycle parameters investigated were; fertilisation success, larval survival, sperm motility (percent motility and curvilinear velocity) and sperm DNA damage. Exposure to decadienal and copper sulphate in isolation resulted in dose- and time-dependent reductions for each evaluated endpoint. Fertilisation success was heavily impacted at concentrations of up to 10µM for both compounds. Copper sulphate was more toxic in larval survival assays. Sperm motility impacts, although variable, exhibited rapid onset with pronounced reductions in sperm swimming performance observed within 3min of exposure. The extent of DNA damage was dose-dependent, and in the case of decadienal, rapid in onset. Dual compound exposures resulted in enhanced overall toxicity in all assays. Logistic regression analysis of fertilisation and larval survival assays showed significant synergistic interactions between decadienal and copper sulphate; an increase in concentration of either compound resulted in enhanced toxicity of the other. Longer exposure durations during larval survival assays demonstrated a further increase in both toxicity and synergism. The results indicate that the effects of additional environmental stressors must be considered when attempting to extrapolate laboratory-derived single compound exposures to field situations.