Effective biomass harvesting of marine diatom Chaetoceros muelleri by chitosan-induced flocculation,preservation of biomass,and recycling of culture medium for aquaculture feed application

Microalgae are a promising new source of biomass; however, large-scale economical harvesting of microalgal biomass is a major technological and economic challenge, limiting the commercial production of microalgal biomass for high-value compounds. In this study, the cationic polymer chitosan was used for the harvesting of the marine diatom Chaetoceros muelleri. Natural flocculation, and pH and chitosan-induced flocculation were studied in detail. The effects of flocculant dosage, culture pH, initial biomass concentration, and sedimentation time were investigated on biomass recovery. The results showed that flocculation efficiency can reach > 99% with an optimum dosage of chitosan (80 mg L^?1) at pH 9.6 and settling time of 40 minutes for biomass concentration from 0.2 to 1.2 g L^?1. The reusability of the recycled water, preservation of biomass after harvesting, and cost of the harvesting process were evaluated. The results showed that the chitosan-induced flocculation offers an efficient, cost-effective, rapid, and sustainable harvesting method for C. muelleri biomass for food and feed applications in aquaculture.

     

Effects of kinetics of adsorption and coalescence on continuous foam concentration of proteins: Comparison of experimental results with model predictions

Protein enrichment and recovery were measured in a continuous foam concentration column for bovine serum albumin (BSA) for different pool heights, foam heights, superficial gas velocities, bubble sizes, feed flow rates, pH, and ionic strengths. Protein enrichment was found to decrease with an increase in pool height for low pool heights, reach a minimum at an intermediate pool height, and subsequently increase with pool height for sufficiently large pool heights eventually approaching an asymptotic value. Such a behavior was due to the combined effects of kinetics of adsorption of protein and coalescence. The increase in protein enrichment with pool height was due to the predominant effect of kinetics of adsorption of protein, whereas the opposite behavior at low pool heights was due to the predominant effect of coalescence in the foam. Protein enrichment was found to be higher for smaller feed concentrations, smaller gas velocities, larger bubble sizes, and larger foam heights. Enrichment at pH values different from the isoelectric point was found to be higher because of more coalescence. A model for foam concentration of proteins was employed to predict enrichment and recovery. The model predictions agreed well with the experimental data. (c) 1996 John Wiley & Sons, Inc.     

Isolation and purification of protease from human placenta by foam fractionation

The effect of operating parameters like pH, protein concentration, column geometry, and gas flow rate on the separation efficiency of proteolytic enzymes from crude human placental homogenate has been studied in a batch foam column. Purification has been found to be optimum at pH 8.0, close to the isoelectric pH, at which the surface adsorption of the protein on the foam bubbles is maximum. Both purification and recovery varied significantly with total protein concentration. Stable bubble formation was hindered at lower protein concentrations, while extraneous proteins rather than the protease were preferentially adsorbed at higher protein concentrations, decreasing the purification efficiency. Column diameter and column height should be optimized for any specific feed protein concentration and gas flow rate. However, the enrichment ratio was found to decrease with the increase in flow rate. The results indicate that foam fractionation is an effective separation process for recovering valuable biochemicals from biological materials.     

Foam fractionation of globular proteins

Foam fractionation of bovine serum albumin (BSA) was studied as a model system for potato wastewater. The effects of feed concentration, superficial gas velocity, feed flow rate, bubble size, pH, and ionic strength on the enrichment and recovery of BSA were investigated in a single-stage continuous foam fractionation column. Enrichments ranged from 1.5 to 6.0 and recoveries from 5 to 85%. The feed concentrations were varied from 0.01 to 0.2 wt %, and enrichments were found to increase with lower feed concentrations. Enrichments also increased with lower superficial gas velocities and larger bubble sizes. At sufficiently low feed flow rates, enrichment was found to increase with an increase in the flow rate, eventually becoming insensitive to the feed flow rate at higher values. The pH was varied from 3.5 to 7.0 and ionic strength from 0.001M to 0.2M. The effects of pH and ionic strength were found to be coupled with bubble size. A minimum bubble size was found at pH 4.8, the isoelectric point of BSA, resulting in a minimum in the enrichment. Bubble size, and thus enrichment, was found to increase as the ionic strength decreased from 0.2M to 0.01M. Previous models(1,2) for the hydrodynamics of foam column were extended for a singlestage continuous foam fractionation column for the prediction of enrichment and recovery. The model assumed adsorption equilibrium, infinite surface viscosity, and bubbles of the same size. Though coalescence was formally accounted for in the model by considering bubble size as a function of foam height, calculations for the experimental runs were performed only for the case of no coalescence. Quantitative predictions of enrichment and recovery could not be made with a single representative bubble size because of the broad inlet bubble size distribution as well as broadening of the distribution as a result of coalescence. The experimental enrichments were higher and recoveries were lower than the model predictions, the discrepancy being more pronounced at lower feed concentrations because of increased coalescence. The higher enrichments are due to the predominant effect of internal reflux as a result of coalescence whereas the lower recoveries are a result of detrimental effects of broadening bubble size distributions.     

Performance of a continuous foam separation column as a function of process variables

Enrichment and recovery of bovine serum albumin has been examined in a continuous foam separation column. The effects of the operating factors, superficial air velocity, feed flow rate, feed concentration and pH on the above characteristics was investigated. The protein enrichment decreased with the increase in the value of each of these parameters. Protein recovery increased with increasing air velocity, decreased with increasing feed flow rate and did not change very much with increasing feed concentration. Maximum protein recovery was obtained at the isoelectric point (pH 4.8) of the protein. Maximum protein recovery was found to be a strong function of the air velocity in the range 0.05-0.15 cm/s. Further increase in air velocity did not have much effect on recovery because of very large bubbles formed as a result of coalescence. Bubble size was determined as a function of the above factors in the liquid and foam sections of the column. It was found to be dependent on protein concentration, feed flow rate and solution pH. The effect was more significant in the foam section of the column. The bubbles in the foam section were significantly larger (about 3-10 times) than those in the liquid, with a sharp change at the foam-liquid interface. The bubble size measurements were used to calculate the interfacial area and it was shown that the rate of protein removal increases with increasing interfacial area.     

Cell recovery by continuous flotation

Summary Cell recovery by means of continuous flotation of the Hansenula polymorpha cultivation medium without additives was investigated as a function of the cultivation conditions as well as of the flotation equipment construction and flotation operational parameters. The cell enrichment and separation is improved at high liquid residence times, high aeration rates, small bubble sizes, increasing height of the aerated column, and diameter of the foam column. Increasing cell age and cultivation with nitrogen limitation reduce the cell separation.Symbols C_P cell mass concentration in medium g·l^–1 - C_R cell mass concentration in residue g·l^–1 - C_S cell mass concentration in foam liquid g·l^–1 - V equilibrium foam volume cm³ - V gas flow rate through the aerated liquid column cm³·s^–1 - V_F feed rate to the flotation column ml/min - ¹ V _S/V foaminess s - mean liquid residence time in the column s     

Comparative life cycle assessment of a commercial algal multiproduct biorefinery and wild caught fishery for small pelagic fish

Purpose

The purpose of this study was to compare the environmental impacts of omega-3 fatty acid (n-3), high protein feed and biofuel production from algae to the impacts of the production of those products from fish.

Methods

The functional unit was the production of one metric ton of omega-3 fatty acids from algae (fish) and the accompanying co-products of biofuel and high protein feed. This was a cradle to gate LCA. Four scenarios were used in this model. The algae multiproduct model (MPM) scenario was the baseline using only unit operations currently in use at the reference facility (Cellana LLC). A low-energy centrifuge replaced the existing conventional centrifuge (MPM (LE)) to reduce energy consumption. The MPM was improved in a different manner, employing membrane filtration prior to centrifugation (MPM (MF)). These three scenarios were compared to the conventional production of the same products from fish (conventional product model: CPM). This life cycle assessment investigated the following impacts: ozone depletion potential, global warming potential, smog formation potential, acidification potential, and eutrophication potential.

Results and discussion

The environmental impacts of producing omega-3 fatty acids from algae were higher than producing omega-3 fatty acids from fish if membrane filtration was not used. Membrane filtration reduced most of the environmental impacts of the algae system by more than 50%. Fuel consumption was the only factor that caused the fish systems to change by greater than 10% from the baseline. Productivity, membrane filtration electricity, and annual operating days could each affect the environmental impacts of the algae system by greater than 10% from the baseline. Improvements to the algae system depend on improvements to cultivation and harvesting, with the impacts from processing being very small.

Conclusions

This study presented results comparing the environmental impacts from a multiproduct system from algae and from fish. The results of this study can serve as a benchmark for the environmental impacts of an algal multiproduct biorefinery compared to the conventional production of those same products from fish. Areas of improvement have been identified for the algae production system for dewatering and cultivation. The amount of n-3 had little impact on the n-3 market but had a significant effect on the existing algal n-3 market. The amount of fuel and feed produced had a negligible effect on both markets.

     

Enrichment in axial direction of aqueous foam in continuous foam separation

Estimation of overhead production enrichment in continuous foam separation was conducted with a surfactant: sodium n-dodecylbenzenesulfonate (SDBS) and soluble proteins: ovalbumin (OA) and hemoglobin (HB). Axial profiles of the volumetric flow rate and the concentration of the collapsed foam liquid within the column were measured, and the enrichment ratio and the liquid holdup in axial direction were determined experimentally. The proposed model was fitted to the experimental results obtained with various experimental conditions (superficial gas velocity, feed concentration and pH) and was in reasonable agreement with the experimental data by using the least square regression. The present model makes it possible to estimate the foamate concentration at a desired foam height.     

The composition and structure of photosystem I‐associated antenna from Cyanidioschyzon merolae

Red algae contain two types of light‐harvesting antenna systems, the phycobilisomes and chlorophyll a binding polypeptides (termed Lhcr), which expand the light‐harvesting capacity of the photosynthetic reaction centers. In this study, photosystem I (PSI) and its associated light‐harvesting proteins were isolated from the red alga Cyanidioschyzon merolae. The structural and functional properties of the largest PSI particles observed were investigated by biochemical characterization, mass spectrometry, fluorescence emission and excitation spectroscopy, and transmission electron microscopy. Our data provide strong evidence for a stable PSI complex in red algae that possesses two distinct types of functional peripheral light‐harvesting antenna complex, comprising both Lhcr and a PSI‐linked phycobilisome sub‐complex. We conclude that the PSI antennae system of red algae represents an evolutionary intermediate between the prokaryotic cyanobacteria and other eukaryotes, such as green algae and vascular plants.     

Technology of streptomycin sulfate separation by two-stage foam separation

Industrial discharges from manufacturing streptomycin sulfate (SS) are inhibitory to biological wastewater treatment and need to be stripped of residual SS. For effective SS recovery from the wastewater, a two-stage foam separation technology was investigated using a column with a vertical ellipsoid-shaped channel (VEC) and a conventional one, and sodium dodecyl sulfate (SDS) served as the collector. The mechanism of enhancing foam drainage by VEC was theoretically analyzed. In the first stage, the column with VEC was used and under the optimal conditions of the liquid-loading volume 300 mL, volumetric airflow rate 100 mL/min, the initial pH 7.0 and the molar ratio of SDS to SS 8.0, an improved SS enrichment ratio of 16.7 was obtained. In the second stage, a conventional column was used and with a volumetric airflow rate of 450 mL/min, the foamate had a SS concentration of about 0.5 g/L, so it was used as the feed solution of the first stage. By the two-stage technology, the total SS recovery percentage reached as high as 99.7%. Thus, it was significantly effective for the two-stage foam separation technology to recover SS from the simulative wastewater.     

Continuous foaming for protein recovery: part I. Recovery of beta-casein

Foam separation is known to have potential for separation of biological molecules with a range of surface activities. A statistical study (factorial design) was carried out to establish the optimum operating conditions for the continuous foam separation of beta-casein. Maximum values of enrichment of beta-casein into the foam phase were found for low levels of initial feed protein concentration, gas flow rate, feed-flow rate, and high foam heights. Maximum values of protein recovery, were generally found at high levels of initial feed protein concentration, gas-flow rate, feed-flow rate, and low foam heights. The highest values obtained for enrichment and separation ratio were 54.7 and 181.3, respectively, with a simultaneous protein recovery of 62%; thus, illustrating the potential effectiveness of this technique. The effect of foaming on protein conformation is also important, and in this study protein structure was analyzed before and after foam separation experiments. Techniques used were: native polyacrylamide gel electrophoresis (PAGE), UV absorbance spectroscopy, circular dichroism, and fluorescence. Native PAGE showed no detectable changes in protein structure. However, absorbance scanning, fluorimetry, and circular dichroism revealed some conformational changes over a range of concentration effects.     

Effect of cereal harvesting method on a recent population decline of Corn Buntings Emberiza calandra on the Western Isles of Scotland

Capsule Population decline since 1995 is associated with the harvesting of cereals as arable silage.

Aim To survey Corn Bunting populations on the Western Isles and test whether population changes since 1995 are associated with change in cereal harvesting methods.

Methods Twelve areas of machair on the Uists and Benbecula were re-surveyed in 2002, after previous surveys in 1983 and 1995. These and other areas of suitable habitat on these islands, plus Berneray, Baleshare, Vallay, Barra and Vatersay, were surveyed in 2003–05 to provide a breeding population estimate for the Western Isles. Cereal strips in the 12 core survey areas were mapped in 2002 and the timing and method of harvesting of each was recorded. Cattle-feeding stands in 24 crofting townships were visited in March 2003 to record the type of fodder available, measure grain abundance and ripeness in fodder samples, and record foraging Corn Buntings. Experimental trials tested whether Corn Buntings selected fodder with riper grain.

Results The Western Isles Corn Bunting population was estimated at 117 occupied territories in 2005, a decline of 17% since 2002/03, and of at least 62% since 1995. Population decline was greater in areas with higher proportions of the cereal area harvested as arable silage. Arable silage contained fewer and less-ripe grains than traditionally harvested cereal, and was less likely to attract feeding Corn Buntings. Corn Buntings preferred to feed on silage containing riper grain.

Conclusions The remaining Corn Bunting population of the Western Isles is declining rapidly, probably because of reduction in winter grain supply caused by change from traditional reaper-binder harvesting and stacking of fully ripe cereals to harvesting the crop as unripe or partially ripe arable silage. Management change to restore a supply of ripe grain over winter is likely to be essential if this population decline is to be halted and reversed.     

Sulphur stable isotopes can distinguish trophic dependence on sediments and plankton in boreal lakes

1. Stable isotopes of carbon are useful for differentiating between freshwater food chains based on planktonic algae or benthic algae, but are reported to be of limited use for identifying food chains based on sedimentary detritus. Because data from marine systems suggest that stable isotopes of sulphur (δ³⁴S values) have potential in this regard, we tested their utility in freshwater lakes.
2. We found that sulphate in the water column of four boreal lakes was enriched in ³⁴S compared to the sulphur in bulk sediments from these lakes. Furthermore, within a given lake, insects known to feed on sediment (directly or via predation) had δ³⁴S values similar to those of sediment, whereas planktonic and benthic invertebrates known to feed on suspended particles had δ³⁴S values similar to those of sulphate in the water column.
3. Using the stable S isotope values of invertebrates that obtain their S from either the sediment or the water column as end members in a two-source mixing model, we show that two fish species obtain their food from both planktonic and sedimentary sources. Furthermore, model results suggest that, as expected, the more benthic-feeding fish species obtains more of its S from the sediment compartment than does the species that feeds in the water-column.
4. Our results suggest that measurements of stable sulphur isotopes provide a means of distinguishing between members of food chains that are based in the water column from those based on sedimentary detritus. As such, they would be a useful complement to stable C isotopes that are used to distinguish between food chains based on planktonic or benthic algae.     

Foam separation of microbial cells

Batch foam separation has been employed to separate Saccharomyces carlsbergensis cells from their broth without the use of any external surface-active agent. A model has been developed to predict the foamate cell concentration as well as the variation of cell concentration in the bulk liquid in the foam column as a function of time. The model assumes a linear equilibrium relation between the cell concentrations at the interface and the bulk. The foam has interface as well as interstitial liquid. The interface is assumed to be in equilibrium with the interstitial liquid, which in turn is assumed to have the same concentration as the bulk. The interfacial area is calculated by assuming the foam bubbles to be pentagonal dodecahedral in shape. The model has been found to explain the results of foam separation of cells quite well, particularly with respect to the effect of bubble size and aeration rate.     

Use of Algae for Removing Heavy Metal Ions From Wastewater: Progress and Prospects

ABSTRACT

Many algae have immense capability to sorb metals, and there is considerable potential for using them to treat wastewaters. Metal sorption involves binding on the cell surface and to intracellular ligands. The adsorbed metal is several times greater than intracellular metal. Carboxyl group is most important for metal binding. Concentration of metal and biomass in solution, pH, temperature, cations, anions and metabolic stage of the organism affect metal sorption. Algae can effectively remove metals from multi-metal solutions. Dead cells sorb more metal than live cells. Various pretreatments enhance metal sorption capacity of algae. CaCl₂ pretreatment is the most suitable and economic method for activation of algal biomass. Algal periphyton has great potential for removing metals from wastewaters. An immobilized or granulated biomass-filled column can be used for several sorption/desorption cycles with unaltered or slightly decreased metal removal. Langmuir and Freundlich models, commonly used for fitting sorption data, cannot precisely describe metal sorption since they ignore the effect of pH, biomass concentration, etc. For commercial application of algal technology for metal removal from wastewaters, emphasis should be given to: (i) selection of strains with high metal sorption capacity, (ii) adequate understanding of sorption mechanisms, (iii) development of low-cost methods for cell immobilization, (iv) development of better models for predicting metal sorption, (v) genetic manipulation of algae for increased number of surface groups or over expression of metal binding proteins, and (vi) economic feasibility.     

Light regulation of light‐harvesting antenna size substantially enhances photosynthetic efficiency and biomass yield in green algae†

One of the major factors limiting biomass productivity in algae is the low thermodynamic efficiency of photosynthesis. The greatest thermodynamic inefficiencies in photosynthesis occur during the conversion of light into chemical energy. At full sunlight the light‐harvesting antenna captures photons at a rate nearly 10 times faster than the rate‐limiting step in photosynthetic electron transport. Excess captured energy is dissipated by non‐productive pathways including the production of reactive oxygen species. Substantial improvements in photosynthetic efficiency have been achieved by reducing the optical cross‐section of the light‐harvesting antenna by selectively reducing chlorophyll b levels and peripheral light‐harvesting complex subunits. Smaller light‐harvesting antenna, however, may not exhibit optimal photosynthetic performance in low or fluctuating light environments. We describe a translational control system to dynamically adjust light‐harvesting antenna sizes for enhanced photosynthetic performance. By expressing a chlorophyllide a oxygenase (CAO) gene having a 5′ mRNA extension encoding a Nab1 translational repressor binding site in a CAO knockout line it was possible to continuously alter chlorophyll b levels and correspondingly light‐harvesting antenna sizes by light‐activated Nab1 repression of CAO expression as a function of growth light intensity. Significantly, algae having light‐regulated antenna sizes had substantially higher photosynthetic rates and two‐fold greater biomass productivity than the parental wild‐type strains as well as near wild‐type ability to carry out state transitions and non‐photochemical quenching. These results have broad implications for enhanced algae and plant biomass productivity.     

High pH-induced flocculation of marine <Emphasis Type="Italic">Chlorella</Emphasis> sp. for biofuel production

Microalgae are being considered as a promising raw material for biofuel production. However, rapid, efficient, and economic technologies for harvesting microalgae are essential for successful applications. In this study, the high–pH-induced flocculation method was applied to harvest marine Chlorella sp. strains. These algae could be concentrated up to approximately 20-fold by increasing pH using NaOH, with a flocculation efficiency of 90 %. When NaOH dosage was low (1 or 3 mM), the flocculation efficiency decreased considerably with the increase of biomass concentration. At higher NaOH dosage tested (5 or 7 mM), flocculation occurred quickly and efficiently, which tended to be independent of biomass concentration. In larger volumes, all strains were flocculated with similar efficiencies (approximately 90 %) after adding 5 mM NaOH. After flocculation, the flocculated algae cells could be re-cultured as inoculum, and the growth yields in flocculated medium were slightly higher than those from fresh medium. Additionally, for each strain, there were no significant differences in lipid extraction yield and fatty acid composition according to different harvesting methods. These results showed that the high–pH-induced flocculation method could be used to harvest marine Chlorella sp. for biofuel production successfully.     

Respiratory relationships of a symbiotic algal-bacterial culture for wastewater nutrient removal

A continuous symbiotic algal-bacterial system was developed consisting essentially of a mixed Chlorella-activated sludge culture which would efficiently remove nutrients from wastewater under aerobic conditions without supplementary aeration. Oxygen decline data were fitted to a mathematical model used to predict respiratory rates, photosynthetic oxygenation, and steady-state oxygen concentrations. Stable relative biological populations and a dissolved oxygen concentration of about 2 mg/1 were maintained during steady-state operation with daily harvesting of excess biomass. Respiratory and physiological relationships indicated that the carbon dioxide-oxygen balance is a primary control that governs the steady-state operation of a symbiotic algal-bacterial culture. The close association of the algae and bacteria resulted in an algal-bacterial floc with settled rapidly yielding a clear supernatant.     

Polarized light based scheme to monitor column performance in a continuous foam fractionation column

Background  

A polarized light scattering technique was used to monitor the performance of a continuously operated foam fractionation process. The S ₁₁ and S ₁₂ parameters, elements of the light scattering matrix, combined together (S ₁₁ +S ₁₂) have been correlated with the bubble size and liquid content for the case of a freely draining foam. The performance of a foam fractionation column is known to have a strong dependence on the bubble size distribution and liquid hold up in foam. In this study the enrichment is used as a metric, representative of foam properties and column performance, and correlated to the S ₁₁ +S ₁₂ parameter.     

Evolution of o(2) in brown algal chloroplasts

A method is described for the isolation of photosynthetically active chloroplasts from four species of brown algae: Fucus vesiculosis, Nereocystis luetkeana, Laminaria saccharina, and Macrocystis integrifolia. When compared to lettuce and spinach chloroplasts, the algal chloroplasts all showed lower activities for both photosystems II and I. Chloroplasts from all the plants produced H₂O₂, with photosystem I functioning as the O₂ reductant in the light. In contrast to the green plants, however, brown algal chloroplasts strongly reduced O₂ under conditions where both photosystems II and I remain active. Relative variable fluorescence values were lower both in intact plants and chloroplasts of the brown algae than for either spinach or lettuce. It is suggested that although light harvesting activities appear similar in all the plants, details of electron transport in brown algae may differ from those of green plants.