Correction: Land-based drip-irrigated culture of Ulva compressa: the effect of culture platform design and nutrient concentration on biomass production and protein content.

[This corrects the article DOI: 10.1371/journal.pone.0199287.].

Simultaneous quantum yield measurements of carbon uptake and oxygen evolution in microalgal cultures.

The photosynthetic quantum yield (Φ), defined as carbon fixed or oxygen evolved per unit of light absorbed, is a fundamental but rarely determined biophysical parameter. A method to estimate Φ for both net carbon uptake and net oxygen evolution simultaneously can provide important insights into energy and mass fluxes. Here we present details for a novel system that allows quantification of carbon fluxes using pH oscillation and simultaneous oxygen fluxes by integration with a membrane inlet mass spectrometer. The pHOS system was validated using Phaeodactylum tricornutum cultured with continuous illumination of 110 µmole quanta m-2 s-1 at 25°C. Furthermore, simultaneous measurements of carbon and oxygen flux using the pHOS-MIMS and photon flux based on spectral absorption were carried out to explore the kinetics of Φ in P. tricornutum during its acclimation from low to high light (110 to 750 µmole quanta m-2 s-1). Comparing results at 0 and 24 hours, we observed strong decreases in cellular chlorophyll a (0.58 to 0.21 pg cell-1), Fv/Fm (0.71 to 0.59) and maximum ΦCO2 (0.019 to 0.004) and ΦO2 (0.028 to 0.007), confirming the transition toward high light acclimation. The Φ time-series indicated a non-synchronized acclimation response between carbon uptake and oxygen evolution, which has been previously inferred based on transcriptomic changes for a similar experimental design with the same diatom that lacked physiological data. The integrated pHOS-MIMS system can provide simultaneous carbon and oxygen measurements accurately, and at the time-resolution required to resolve high-resolution carbon and oxygen physiological dynamics.

Land-based drip-irrigated culture of Ulva compressa: The effect of culture platform design and nutrient concentration on biomass production and protein content.

This work developed a laboratory prototype methodology for cost-effective, water-sparing drip-irrigation of seaweeds, as a model for larger-scale, on-land commercial units, which we envision as semi-automated, inexpensive polyethylene sheet-covered bow-framed greenhouses with sloping plastic covered floors, water-collecting sumps, and pumped recycling of culture media into overhead low-pressure drip emitters. Water droplets form on the continually wetted interior plastic surfaces of these types of greenhouses scattering incoming solar radiation to illuminate around and within the vertically-stacked culture platforms. Concentrated media formulations applied through foliar application optimize nutrient uptake by the seaweeds to improve growth and protein content of the cultured biomass. An additional attribute is that seaweed growth can be accelerated by addition of anthropogenic CO2-containing industrial flue gases piped into the head-space of the greenhouse to reuse and recycle CO2 into useful algal biomass. This demonstration tested three different drip culture platform designs (horizontal, vertical and slanted) and four increasing fertilizer media concentrations (in seawater) for growth, areal productivity, and thallus protein content of wild-collected Ulva compressa biomass, against fully-submerged controls. Cool White fluorescent lights provided 150-200 µmol photon m-2 s-1 illumination on a 12/12 hr day/night cycle. Interactive effects we tested using a four-level single factorial randomized block framework (p<0.05). Growth rates and biomass of the drip irrigation designs were 3-9% day-1 and 5-18 g m-2 day-1 (d.w.) respectively, whereas the fully-submerged control group grew better at 8-11% per day with of 20-30 g m-2 day-1, indicating further optimization of the drip irrigation methodology is needed to improve growth and biomass production. Results demonstrated that protein content of Ulva biomass grown using the vertically-oriented drip culture platform and 2x fertilizer concentrations (42:16:36 N:P:K) was 27% d.w., approximating the similarly-fertilized control group. The drip methodology was found to significantly improve gas and nutrient mass transfer through the seaweed thalli, and overall, the labor- and-energy-saving methodology would use a calculated 20% of the seawater required for conventional on-land tank-based tumble culture.

System and method for research-scale outdoor production of microalgae and cyanobacteria.

Eukaryotic microalgae and cyanobacteria have recently reemerged as promising organisms in the effort to develop sustainable options for production of food and fuel. However, substantial discrepancies consistently arise between laboratory and outdoor cultivation, and gains demonstrated using laboratory technologies have not paralleled gains observed in field demonstrations. For these reasons, a low-maintenance system and process for research-scale outdoor cultivation of a variety of both freshwater and marine microalgae and cyanobacteria was developed. Nine genera were evaluated in the system, demonstrating cultivation of both laboratory model and commercial-production organisms. Hundreds to thousands of grams of dry biomass could be produced in a single growth cycle, suitable for a variety of uses including inoculum generation, protein production, and biofuel applications. Following testing in outdoor stock-ponds, Scenedesmus and Nannochloropsis were grown semi-continuously in an 8000 L airlift-driven raceway, yielding in total over 8 kg of dry biomass for each strain.

Life cycle GHG emissions from microalgal biodiesel--a CA-GREET model.

A life cycle assessment (LCA) focused on greenhouse gas (GHG) emissions from the production of microalgal biodiesel was carried out based on a detailed engineering and economic analysis. This LCA applies the methodology of the California Low Carbon Fuel Standard (CA LCFS) and uses life cycle inventory (LCI) data for process inputs, based on the California-Modified Greenhouse Gases, Regulated Emissions, and Energy use in Transportation (CA GREET) model. Based on detailed mass and energy balances, calculated GHG emissions from this algal biodiesel system are 70% lower than those of conventional diesel fuel, meeting the minimum 50% GHG reduction requirements under the EPA RFS2 and 60% for the European Union Renewable Energy Directive. This LCA study provides a guide to the research and development objectives that must be achieved to meet both economic and environmental goals for microalgae biodiesel production.

Environmental flow regimes for Dysidea avara sponges.

The aim of our research is to design tank systems to culture Dysidea avara for the production of avarol. Flow information was needed to design culture tanks suitable for effective production. Water flow regimes were characterized over a 1-year period for a shallow rocky sublittoral environment in the Northwestern Mediterranean where D. avara sponges are particularly abundant. Three-dimensional Doppler current velocities at 8-10-m depths ranged from 5 to 15 cm/s over most seasons, occasionally spiking to 30-66 cm/s. A thermistor flow sensor was used to map flow fields in close proximity ( approximately 2 cm) to individual sponges at 4.5-, 8.8-, and 14.3-m depths. These "proximal flows" averaged 1.6 cm/s in calm seas and 5.9 cm/s during a storm, when the highest proximal flow (32.9 cm/s) was recorded next to a sponge at the shallowest station. Proximal flows diminished exponentially with depth, averaging 2.6 cm/s +/- 0.15 SE over the entire study. Flow visualization studies showed that oscillatory flow (0.20-0.33 Hz) was the most common regime around individual sponges. Sponges at the 4.5-m site maintained a compact morphology with large oscula year-around despite only seasonally high flows. Sponges at 8.8 m were more erect with large oscula on tall protuberances. At the lowest-flow 14.3-m site, sponges were more branched and heavily conulated, with small oscula. The relationship between sponge morphology and ambient flow regime is discussed.

Hypothesized kinetic models for describing the growth of globular and encrusting demosponges.

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

Large-scale production of pharmaceuticals by marine sponges: sea, cell, or synthesis?

Marine sponges are known to produce an overwhelming array of secondary metabolites with pharmaceutical potential. The technical and economical potential of using marine sponges for large-scale production of these compounds was assessed for two cases: the anticancer molecule halichondrin B from a Lissodendoryx sp., and avarol from Dysidea avara for its antipsoriasis activity. An economic and technical analysis was done for three potential production methods: mariculture, ex situ culture (in tanks), and cell culture. We concluded that avarol produced by mariculture or ex situ culture could become a viable alternative to currently used pharmaceuticals for the treatment of psoriasis. Production of halichondrin B from sponge biomass was found to not be a feasible process, mainly due to the extremely low concentration of the compound in the sponge. Technical feasibility was also analyzed for five alternatives: chemical synthesis, wild harvest, primmorph culture, genetic modification and semi-synthesis. It was concluded that the latter two approaches could prove to be valuable methods for the production of pharmaceuticals, based on chemical structures of secondary metabolites present in trace amounts in marine sponges.

Aquaculture of three phyla of marine invertebrates to yield bioactive metabolites: process developments and economics.

Large-scale, renewable supplies of chemical constituents derived from marine invertebrates have limited development of potential new natural product drugs. This paper describes the development of two in-sea aquaculture systems designed and engineered for production of large quantities of biomass for two species of marine invertebrates desired for their natural product chemical constituents. The two invertebrates and their products were: (1) the cosmopolitan, arborescent bryozoan Bugula neritina (Phylum Bryozoa) for its anticancer chemical constituent bryostatin 1; and (2) Ecteinascidia turbinate (Phylum Tunicata) the source of anticancer ecteinascidin 743. For the third invertebrate Phylum Porifera, and its representative sponge Acanthella cavernosa (desired for its anti-parasitic and anti-infective kalihinols) in-sea systems were not developed in favor of controlled environment tank aquaculture systems. For the bryozoan and tunicate, projected economics for commercial-scale in-sea production proved cost effective. This was in contrast to the controlled environment sponge culture tank system, which did not prove to be economical due to inherent slow growth and low natural product yields of the sponge in culture. A non-destructive method for "milking" natural product chemicals from sponges was tested and is described.