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In the monitoring the discharge of ballast water, the count of living algal cells is of utmost significant. Variable fluorescence, denoted as Fv, stands as an optimal parameter for photosynthetic fluorescence, efficiently charactering the living algal cells count, unaffected by the ballast waters' complex background fluorescence environment. This study deeply investigates the quantitative relationship between Fv and the count of living algal cells. Observations indicate that single cell fluorescence yield (abbreviated as SCF) varies significantly across different algae species, leading to considerable errors in quantifying living algal cell count in ballast water with unknown components using the calibration relationship between Fv and the cell count. Thus, correcting SCF prior to calibration becomes necessary. The paper proposes an innovative SCF correction method based on cell cross-sectional area and an e μ factor (where μ is the expected value of the functional absorption cross-section of PSII) This method mitigates the influence of cell size and species differences on quantifying the living algal cell count. Correction operation trials revealed that dividing the SCF measurement by cell cross-sectional area and multiplying by e μ enhanced the correction effect. Comparative experiments demonstrated marked improvement: Relative errors (REs) for Chlorella pyrenoidosa and Chlorella marine, both belonging to the Chlorophyta group, fell from 92.1% and 90.6% to 37.2% and 9.5% respectively post-correction. Similarly, REs for Thalassiosira weissflogii and Nitzschia closterium minutissima, from the Bacillariophyta group, decreased from 74.7% and 68.1% to 14.3% and 19.1% respectively. The RE of Peridinium from the Pyrrophyta group dropped from 28.4% to 12.1%. The results underscore the effectiveness of cell cross-sectional area and e μ in correcting SCF, thus offering a novel correction method for swift and precise measurement of living algal cell count in ballast water, based on variable fluorescence., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Membrane lipid remodeling contributes to the environmental acclimation of plants. In the green lineage, a betaine lipid, diacylglyceryl-N,N,N-trimethylhomoserine (DGTS), is included exclusively among green algae and nonflowering plants. Here, we show that the green alga Chlorella kessleri synthesizes DGTS under phosphorus-deficient conditions through the eukaryotic pathway via the ER. Simultaneously, phosphatidylcholine and phosphatidylethanolamine, which are similar to DGTS in their zwitterionic properties, are almost completely degraded to release 18.1% cellular phosphorus, and to provide diacylglycerol moieties for a part of DGTS synthesis. This lipid remodeling system that substitutes DGTS for extrachloroplast phospholipids to lower the P-quota operates through the expression induction of the BTA1 gene. Investigation of this lipid remodeling system is necessary in a wide range of lower green plants for a comprehensive understanding of their phosphorus deficiency acclimation strategies., (© 2022. The Author(s).)

The judgment of microalgae viability is a vital procedure in the process of microalgae culture and treatment, which also plays an important role in bioremediation, bioindication, and pharmacology fields. The current conventional methods for defining living/dead microalgal cells are complicated or laborious. Hence, developing a simple and reliable detection method for microalgae viability is still challenging. Here, we developed chlorella-based carbonized polymer dots (c-CPDs) by a hydrothermal method. Due to their small average size of 5.0 nm, obvious excitation-dependent emission, stable fluorescence properties, and low toxicity, c-CPDs could be used for distinguishing living or dead chlorella by testing different fluorescence characteristics of c-CPD-labeled chlorella. Compared with conventional cellular dyes used for differentiating living/dead microalgae, c-CPDs significantly reduced toxicity, showing good sensitivity and reliability. This work provided a method to prepare environmentally friendly carbon dots (CDs) using microalgae, which had potential to be prepared on a large scale and might be applied feasibly in the preparation of doped CDs by controlling the growth of chlorella.

Main Conclusion: The supramolecular organization of the photosystem supercomplexes in the green alga Chlorella sorokiniana belonging to Trebouxiophyceae are essentially the same as those of Chlamydomonas reinhardtii belonging to Chlorophyceae. The photosynthetic conversion of light energy into chemical energy is performed by photosystems II and I (PSII and PSI) embedded within the thylakoid membranes. In plants and green algae, PSII and PSI comprise the core complex and light-harvesting complexes (LHCII and LHCI), forming PSII-LHCII and PSI-LHCI supercomplexes, respectively. The structural information about photosystem supercomplexes of green algae has been limited to chlorophytic algae. Here, to obtain an insight into the evolution of Chlorophyta, we determined the supramolecular organization of the PSII-LHCII and PSI-LHCI supercomplexes from the freshwater green alga Chlorella sorokiniana, which belongs to Trebouxiophyceae. The obtained results showed that the supramolecular organizations of the photosystem supercomplexes in C. sorokiniana were essentially the same as those of the model green alga C. reinhardtii, which belongs to Chlorophyceae, namely PSII-LHCII supercomplex formed the C 2 S 2 M 2 L 2 configuration and PSI-LHCI supercomplex was associated with 10 LHCI subunits.

Anaerobic digestion of microalgal biomass for biogas production may be limited due to the cell wall resulting in an inefficient bioconversion. Enzymatic pretreatments are applied for inducing cell damage/lysis and organic matter solubilisation and this way increasing biogas production. We evaluated enzymatic pretreatments in different conditions for comparing in relation to cell wall rupture, increase of soluble material and increase in biogas production through anaerobic digestion performance in BMP assay. Chlorella sorokiniana cultures were subjected to three different enzymatic pretreatments, each under four different conditions of enzyme/substrate ratio, pH and application time. The results showed increases over 21% in biogas productions for all enzymatic pretreatments. Enzymatic pretreatment was effective at damaging microalgae cell wall, releasing organic compounds and increasing the rate and final methane yield in BMP tests. We observed a synergistic activity between the mixtures enzymes, which would depend on operational conditions used for each pretreatment.

The frequent occurrence of algal blooms in surface water has attracted more and more attention, which caused many water quality problems, including disinfection byproducts (DBPs). Algal organic matter (AOM) including intracellular organic matter (IOM) and extracellular organic matter (EOM), was a well-known precursor to DBPs formation in drinking water. This study evaluated the effect of ultraviolet (UV) irradiation on the cell integrity, IOM release and DBPs formation during subsequent chlorination of Chlorella sp. Results showed the damage rates of algal cells increased to 40.1% after the high UV irradiation of 528 mJ/cm 2 , which contributed to the release of IOM. In addition, UV irradiation was effective in reducing the formation of haloacetic acids (HAAs) both in AOM and IOM, but promoted the formation of nitrogenous DBPs (N-DBPs) from AOM in subsequent chlorination. Furthermore, neutral pH exerted a positive effect on the formation of DBPs. UV irradiation decreased the bromine substitution factor (BSF) value of AOM at a high bromide level. The BSF values increased with increasing of the concentration of bromide. Moreover, more amino acids and low molecular weight precursors were produced after UV irradiation in filtered supernatant, which contributed to the formation of N-DBPs with algal chlorination. Overall, this information demonstrated pre-oxidation of UV irradiation could be used to treat the algal-rich drinking water., (Copyright © 2019. Published by Elsevier B.V.)

A mild and low-energy cell disruption method with high efficiency has growing application potential in both the extraction of high-value microalgal products and the inactivation of microalgal cells. Conventional technologies available have disadvantages including high energy consumption, the use of chemicals and so on. Here, this study developed an efficient microalgal cell disruption method using the copper oxide nanowire (CuONW)-modified three-dimensional (3D) copper foam electrodes with a low applied voltage. Electrodes with nanowires synthesized at 400 °C, the optimal preparation temperature, achieved efficient microalgal cell electroporation. Microalgal cells were completely inactivated and disrupted at the voltage of 2 V with the hydraulic retention time (HRT) of 10 s. Scanning electron microscopy (SEM) images showed obvious electroporation damage on the cell surface upon electroporation-treatment (2 V, 30 s). The amount of released cellular inclusion increased significantly with prolonged HRT and the energy consumption of this technology was only 0.014 kWh/kg via the treatment of 2 V and 10 s. This study provided a novel, energy-efficient and chemical-free technique for both microalgal products extraction and cell inactivation., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The combined effect of polystyrene (PS) particles and triphenyltin chloride (TPTCl) to the green algae Chlorella pyrenoidosa was studied. The 96 h IC 50 of TPTCl to the green algae C. pyrenoidosa was 30.64 μg/L. The toxicity of PS particles to C. pyrenoidosa was size-dependent, with the 96 h IC 50 at 9.10 mg/L for 0.55 μm PS but no toxicity observed for 5.0 μm PS. The exposure to 0.55 μm PS led to damage on structure of algal cells, which could in turn cause inhibition on photosynthesis and population growth of the green algae. TPTCl concentrations in test medium were lowered by 15-19% at presence of 0.55 μm PS particles, indicating a reduced bioavailability of TPTCl. In spite of this reduced bioavailability, the presence of PS increased the toxicity of TPTCl, which might be attributed to facilitated uptake of TPTCl by the green algae after the damage of cell structure. The overall results of the present study provided important information on the effect of PS on the bioavailability and toxicity of TPTCl to phytoplankton species.

Thorium (Th) is a natural radioactive element present in the environment and has the potential to be used as a clean nuclear fuel. Relatively little is known about the aquatic toxicity of Th, especially in nanoparticulate form, which may be the main chemical species of Th in the natural waters. In this study, impacts of ThO 2 nanoparticles (NPs) with two different sizes (52 ± 5 nm, s-ThO 2 vs. 141 ± 6 nm, b-ThO 2 ) on a green alga Chlorella pyrenoidosa (C. pyrenoidosa) were evaluated. Results indicated that C. pyrenoidosa was more sensitive to s-ThO 2 (96-h EC 30  = 64.1 μM) than b-ThO 2 (96-h EC 30  = 100.2 μM). Exposure to 200 μM of ThO 2 NPs reduced the chlorophyll-a and chlorophyll-b contents of the algal cells. At 96 h, SEM and TEM showed that more agglomerates of s-ThO 2 than those of b-ThO 2 were attached onto the surface of algal cells. Reactive oxygen species (ROS) generation and membrane damage were induced after the attachment of high concentrations of ThO 2 NPs. The heteroagglomeration between ThO 2 NPs and algal cells and increased oxidative stress might play important roles in the toxicity of ThO 2 NPs. To the best of our knowledge, this is the first report on aquatic toxicity of ThO 2 NPs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Organophosphorus compounds exhibit a wide range of toxicity to mammals. In this study the effect of malathion on the growth and biochemical parameters of microalgae was evaluated. Three microalgae (Micractinium pusillum UUIND2, Chlorella singulari UUIND5 and Chlorella sorokiniana UUIND6) were used in this study. Among the three algal strains tested, Chlorella sorokiniana UUIND6 was able to tolerate 100 ppm of malathion. The photosynthetic pigments, the protein, carbohydrate and lipid contents of microalgal cells were also analyzed. About 90% degradation was recorded in 25 ppm, 50 ppm and 70% was recorded in 100 ppm of malathion by Chlorella sorokiniana. A mechanism of degradation of malathion by Chlorella sorokiniana is proposed in this study. Activity of carboxylesterase was increased in algal cells cultivated in malathion containing medium which confirmed that malathion degraded into phosphate. Increased amount of Malondialdehye (MDA) indicate the development of free radicals under the stress of malathion which substantialy increase de novo TAG biosynthesis, while increased level of superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) suggested their association in scavenging of free radical., (Copyright © 2019 Elsevier B.V. All rights reserved.)

There has been a developing technology in algae with pharmaceuticals wastewater. However, the effect and the underlying mechanism of pharmaceuticals on algae are not well understood. To investigate the effect and mechanism of pharmaceuticalson microalgae, four pharmaceuticals of clofibric acid (CLF), ciprofloxacin (CIP), diclofenac (DCF) and carbamazepine (CBZ) on C. pyrenoidosa culture were analyzed. At low concentrations (<10 mg/L), the pharmaceuticals, especially the DCF, exhibited positive effects on both the structure and function of algal cultures; algal growth (i.e., chlorophyll a accumulation, lipid accumulation) and activities of antioxidant enzymes were stimulated. The algal metabolite differences of various DCF concentrations were investigated and a total of 91 substances were identified, whose samples were clustered and clearly separated. The key metabolomics pathway analysis found that the DCF promoted the carbohydrate and fatty acid metabolic pathway in C. pyrenoidosa under relatively low concentrations (<10 mg/L). However, the algae metabolomics pathway was disturbed significantly under the action of a high concentration of DCF (>100 mg/L). The study detected the effects of four pharmaceuticals on C. pyrenoidosa and demonstrated that the usage of metabolomics analysis complemented with DCF could be an effective approach to understand the mechanism of molecular evolution in C. pyrenoidosa for microalgal biomass and bioenergy from wastewater in researches of biological resources.

Microalgae biotechnologies are rapidly developing into new commercial settings. Several high value products already exist on the market, and systems development is focused on cost reduction to open up future economic opportunities for food, fuel and freshwater production. Light is a key environmental driver for photosynthesis and optimising light capture is therefore critical for low cost, high efficiency systems. Here a novel high-throughput screen that simulates fluctuating light regimes in mass cultures is presented. The data was used to model photosynthetic efficiency (PE µ , mol photon -1 m 2 ) and chlorophyll fluorescence of two green algae, Chlamydomonas reinhardtii and Chlorella sp. Response surface methodology defined the effect of three key variables: density factor (D f , 'culture density'), cycle time (t c , 'mixing rate'), and maximum incident irradiance (I max ). Both species exhibited a large rise in PE µ with decreasing I max and a minimal effect of t c (between 3-20 s). However, the optimal D f of 0.4 for Chlamydomonas and 0.8 for Chlorella suggested strong preferences for dilute and dense cultures respectively. Chlorella had a two-fold higher optimised PE µ than Chlamydomonas, despite its higher light sensitivity. These results demonstrate species-specific light preferences within the green algae clade. Our high-throughput screen enables rapid strain selection and process optimisation.

Astaxanthin represents a high-value ketocarotenoid that has been gaining great attention from both the science and public communities in recent years. Microalgae are the primary source of natural astaxanthin. Chlorella zofingiensis, a freshwater and oleaginous green microalga, is capable of growing well photoautotrophically, heterotrophically, and mixotrophically for high-density biomass and astaxanthin production. Here we describe the astaxanthin production pipeline using C. zofingiensis as cell factories under different trophic conditions, including strain preparation, inoculation, and cultivation, biomass harvest and dewatering, and astaxanthin extraction, determination, and quantification.

Objective: To fabricate a novel microbial photobioelectrochemical cell using silicon microfabrication techniques., Results: High-density photosynthetic cells were immobilized in a microfluidic chamber, and ultra-microelectrodes in a microtip array were inserted into the cytosolic space of the cells to directly harvest photosynthetic electrons. In this way, the microbial photobioelectrochemical cell operated without the aid of electron mediators. Both short circuit current and open circuit voltage of the microbial photobioelectrochemical cell responded to light stimuli, and recorded as high as 250 pA and 45 mV, respectively., Conclusion: A microbial photobioelectrochemical cell was fabricated with potential use in next-generation photosynthesis-based solar cells and sensors.

Heavy ion beams have many exciting applications, including radiotherapy of deep-seated tumors and simulation tests of space irradiation for astronauts. These beams often use a feature that concentrates the energy deposition largely along the end of the energy pathway, leading to different distributions of biological effects along the axial direction. Currently, there is relatively little information regarding the radial directional difference of biological effects along the heavy ion paths. This study utilized a filter membrane that was quantatively applied with cells to demonstrate a 3D distribution model of irradiation on biological effects in living organisms. Some results have indicated that there is excitatory effect on the non-irradiated regions with energetic ions, which may give new insights into the distribution of biological effects along the paths of heavy ion beams with mid-high energy.

Background: Microalgae have the potential to rapidly accumulate lipids of high interest for the food, cosmetics, pharmaceutical and energy (e.g. biodiesel) industries. However, current lipid extraction methods show efficiency limitation and until now, extraction protocols have not been fully optimized for specific lipid compounds. The present study thus presents a novel lipid extraction method, consisting in the addition of a water treatment of biomass between the two-stage solvent extraction steps of current extraction methods. The resulting modified method not only enhances lipid extraction efficiency, but also yields a higher triacylglycerols (TAG) ratio, which is highly desirable for biodiesel production., Results: Modification of four existing methods using acetone, chloroform/methanol (Chl/Met), chloroform/methanol/H 2 O (Chl/Met/H 2 O) and dichloromethane/methanol (Dic/Met) showed respective lipid extraction yield enhancement of 72.3, 35.8, 60.3 and 60.9%. The modified acetone method resulted in the highest extraction yield, with 68.9 ± 0.2% DW total lipids. Extraction of TAG was particularly improved with the water treatment, especially for the Chl/Met/H 2 O and Dic/Met methods. The acetone method with the water treatment led to the highest extraction level of TAG with 73.7 ± 7.3 µg/mg DW, which is 130.8 ± 10.6% higher than the maximum value obtained for the four classical methods (31.9 ± 4.6 µg/mg DW). Interestingly, the water treatment preferentially improved the extraction of intracellular fractions, i.e. TAG, sterols, and free fatty acids, compared to the lipid fractions of the cell membranes, which are constituted of phospholipids (PL), acetone mobile polar lipids and hydrocarbons. Finally, from the 32 fatty acids analyzed for both neutral lipids (NL) and polar lipids (PL) fractions, it is clear that the water treatment greatly improves NL-to-PL ratio for the four standard methods assessed., Conclusion: Water treatment of biomass after the first solvent extraction step helps the subsequent release of intracellular lipids in the second extraction step, thus improving the global lipids extraction yield. In addition, the water treatment positively modifies the intracellular lipid class ratios of the final extract, in which TAG ratio is significantly increased without changes in the fatty acids composition. The novel method thus provides an efficient way to improve lipid extraction yield of existing methods, as well as selectively favoring TAG, a lipid of the upmost interest for biodiesel production.

In this study, harvesting efficiency (HE) of bioflocculant (egg shell) was observed with variation in flocculent concentrations (0-100mgL -1 ), temperature (30°C, 35°C 40°C, 45°C and 50°C) and variable contact time (0-50min). It was found maximum (≈95.6%) with 100mgL -1 bioflocculant concentration whereas influence of temperature was also observed with optimized concentration of bioflocculant (100mgL -1 ) at 40°C (≈98.1%) and 50°C (≈99.3%), in 30min of contact time. Significant changes in algal cell structures were also analyzed after exposure to various temperatures with microscopy, SEM (Scanning electron microscopy) and EDS (Energy dispersive X-ray spectroscopy) images with and without bioflocculant. The experimental data was found to be a good fit with pseudo-second order kinetic model. The thermodynamic functions such as ΔG (Gibbs free energy), ΔH (enthalpy), ΔS (entropy) were also determined. The negative value of ΔG and positive value of ΔH and ΔS shows the spontaneous and endothermic nature of flocculation process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Effective enhancement of neutral lipid (especially triacylglycerol, TAG) content in microalgae is an important issue for commercialization of microalgal biorefineries. Pressure is a key physical factor affecting the morphological, physiological, and biochemical behaviors of organisms. In this paper, we report a new stress-based method for induction of TAG accumulation in microalgae (specifically, Chlorella sp. KR-1 and Ch. sp. AG20150) by very-short-duration application of mild pressure. Pressure treatments of 10-15bar for 2h resulted in a considerable, ∼55% improvement of the 10-100g/Lcells' TAG contents compared with the untreated control. The post-pressure-treatment increase of cytoplasmic TAG granules was further confirmed by transmission electron microscopy (TEM). Notwithstanding the increased TAG content, the total lipid content was not changed by pressurization, implying that pressure stress possibly induces rapid remodeling/transformation of algal lipids rather than de novo biosynthesis of TAG., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We have developed a micro electromechanical systems (MEMS) device which enables plasma treatment for cells cultured in media. The device, referred to as the plasma-on-chip, comprises microwells and microplasma sources fabricated together in a single chip. The microwells have through-holes between the microwells and microplasma sources. Each microplasma source is located on the backside of each microwells. The reactive components generated by the microplasma sources pass through the through-holes and reach cells cultured in the microwells. In this study, a plasma-on-chip device was modified for a stable plasma treatment. The use of a dielectric barrier discharge (DBD) technique allowed a stable plasma treatment up to 3 min. The plasma-on-chip with the original electrode configuration typically had the maximum stable operation time of around 1 min. Spectral analysis of the plasma identified reactive species such as O and OH radicals that can affect the activity of cells. Plasma treatment was successfully performed on yeast (Saccharomyces cerevisiae) and green algae (Chlorella) cells. While no apparent change was observed with yeast, the treatment degraded the activity of the Chlorella cells and decreased their fluorescence. The device has the potential to help understand interactions between plasma and cells., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Algal endosymbiosis is widely distributed in eukaryotes including many protists and metazoans, and plays important roles in aquatic ecosystems, combining phagotrophy and phototrophy. To maintain a stable symbiotic relationship, endosymbiont population size in the host must be properly regulated and maintained at a constant level; however, the mechanisms underlying the maintenance of algal endosymbionts are still largely unknown. Here we investigate the population dynamics of the unicellular ciliate Paramecium bursaria and its Chlorella-like algal endosymbiont under various experimental conditions in a simple culture system. Our results suggest that endosymbiont population size in P. bursaria was not regulated by active processes such as cell division coupling between the two organisms, or partitioning of the endosymbionts at host cell division. Regardless, endosymbiont population size was eventually adjusted to a nearly constant level once cells were grown with light and nutrients. To explain this apparent regulation of population size, we propose a simple mechanism based on the different growth properties (specifically the nutrient requirements) of the two organisms, and based from this develop a mathematical model to describe the population dynamics of host and endosymbiont. The proposed mechanism and model may provide a basis for understanding the maintenance of algal endosymbionts., (© 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Endosymbioses are driving forces underlying cell evolution. The endosymbiosis exhibited by Paramecium bursaria is an excellent model with which to study symbiosis. A single-cell microscopic analysis of P. bursaria reveals that endosymbiont numbers double when the host is in the division phase. Consequently, endosymbionts must arrange their cell cycle schedule if the culture-condition-dependent change delays the generation time of P. bursaria. However, it remains poorly understood whether endosymbionts keep pace with the culture-condition-dependent behaviors of P. bursaria, or not. Using microscopy and flow cytometry, this study investigated the life cycle behaviors occurring between endosymbionts and the host. To establish a connection between the host cell cycle and endosymbionts comprehensively, multivariate analysis was applied. The multivariate analysis revealed important information related to regulation between the host and endosymbionts. Results show that dividing endosymbionts underwent transition smoothly from the division phase to interphase, when the host was in the logarithmic phase. In contrast, endosymbiont division stagnated when the host was in the stationary phase. This paper explains that endosymbionts fine-tune their cell cycle pace with their host and that a synchronous life cycle between the endosymbionts and the host is guaranteed in the symbiosis of P. bursaria.

A compartmentalized genome scale metabolic network was reconstructed for Chlorella variabilis to offer insight into various metabolic potentials from this alga. The model, iAJ526, was reconstructed with 1455 reactions, 1236 metabolites and 526 genes. 21% of the reactions were transport reactions and about 81% of the total reactions were associated with enzymes. Along with gap filling reactions, 2 major sub-pathways were added to the model, chitosan synthesis and rhamnose metabolism. The reconstructed model had reaction participation of 4.3 metabolites per reaction and average lethality fraction of 0.21. The model was effective in capturing the growth of C. variabilis under three light conditions (white, red and red+blue light) with fair agreement. This reconstructed metabolic network will serve an important role in systems biology for further exploration of metabolism for specific target metabolites and enable improved characteristics in the strain through metabolic engineering., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This work reports on an efficient microfluidic approach for continuous production of hollow Ca-alginate microfibers with controllable structures and functions. A coaxial microcapillary microfluidic device combined with a rotator is constructed to produce a cylindrical flow jet with four aqueous solutions as templates for continuous fabrication and collection of microfibers. A four-aqueous-phase flow jet with an intermediate buffer flow between the Ca(2+)-containing and alginate-containing flows is used as the template for microfiber fabrication. The buffer flow efficiently controls the diffusion of Ca(2+) into the alginate-containing flow as well as the crosslinking reaction, thus ensuring the continuous fabrication of hollow Ca-alginate microfibers under relatively low flow rates without clogging of the microchannel. The structure of the hollow microfibers can be flexibly adjusted by changing the flow rates and device dimensions. Meanwhile, the continuous fabrication process of the microfibers allows flexible incorporation of a functional component into the sheath flow for functionalization and addition of active substances in the core flow for encapsulation. This is demonstrated by fabricating hollow Ca-alginate microfibers with a wall containing magnetic nanoparticles for magnetic functionalization and with hollow internals containing Chlorella pyrenoidosa cells for confined growth. This work provides an efficient strategy for continuous fabrication of functional hollow Ca-alginate microfibers with controllable structures and functions.

Background: Within the context of microalgal lipid production for biofuels and bulk chemical applications, specialized higher throughput devices for small scale parallelized cultivation are expected to boost the time efficiency of phototrophic bioprocess development. However, the increasing number of possible experiments is directly coupled to the demand for lipid quantification protocols that enable reliably measuring large sets of samples within short time and that can deal with the reduced sample volume typically generated at screening scale. To meet these demands, a dye based assay was established using a liquid handling robot to provide reproducible high throughput quantification of lipids with minimized hands-on-time., Results: Lipid production was monitored using the fluorescent dye Nile red with dimethyl sulfoxide as solvent facilitating dye permeation. The staining kinetics of cells at different concentrations and physiological states were investigated to successfully down-scale the assay to 96 well microtiter plates. Gravimetric calibration against a well-established extractive protocol enabled absolute quantification of intracellular lipids improving precision from ±8 to ±2 % on average. Implementation into an automated liquid handling platform allows for measuring up to 48 samples within 6.5 h, reducing hands-on-time to a third compared to manual operation. Moreover, it was shown that automation enhances accuracy and precision compared to manual preparation. It was revealed that established protocols relying on optical density or cell number for biomass adjustion prior to staining may suffer from errors due to significant changes of the cells' optical and physiological properties during cultivation. Alternatively, the biovolume was used as a measure for biomass concentration so that errors from morphological changes can be excluded., Conclusions: The newly established assay proved to be applicable for absolute quantification of algal lipids avoiding limitations of currently established protocols, namely biomass adjustment and limited throughput. Automation was shown to improve data reliability, as well as experimental throughput simultaneously minimizing the needed hands-on-time to a third. Thereby, the presented protocol meets the demands for the analysis of samples generated by the upcoming generation of devices for higher throughput phototrophic cultivation and thereby contributes to boosting the time efficiency for setting up algae lipid production processes.

The polarities of the cytoplasm and reactants were measured through dielectric spectroscopy, contact angle test, NMR, and FTIR to investigate the mechanisms underlying acid-catalyzed lipid transesterification in wet microalgal cells subjected to microwave irradiation. Organics with apolar functional groups in the cytoplasm decreased the contact angle of methanol against triglyceride by 13.92°, which subsequently increased transesterification efficiency by 2.4 times. The microalgal biomass, given its higher hydrophilicity index of 1.96 than lipids, was more accessible to hydrophilic alcohols, which subsequently promoted transesterification. Water in the cytoplasm promoted the dielectric constant of methanol and increased the contact angle of methanol against triglyceride by 20.51°, which subsequently decreased transesterification efficiency by 72.6%. The inhibitory effect of water on transesterification weakened with the prolonged carbon lengths of the alcohols because of decreased polarity. Microwave decreased the electric constants of alcohols and reduced the polarity difference between alcohols and lipids, thereby improving transesterification efficiency., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The ability to image large numbers of cells at high resolution enhances flow cytometric analysis of cells and cell populations. In particular, the ability to image intracellular features adds a unique aspect to analyses, and can enable correlation between molecular phenomena resulting in alterations in cellular phenotype. Unicellular microalgae are amenable to high-throughput analysis to capture the diversity of cell types in natural samples, or diverse cellular responses in clonal populations, especially using imaging cytometry. Using examples from our laboratory, we review applications of imaging cytometry, specifically using an Amnis(®) ImageStream(®)X instrument, to characterize photosynthetic microalgae. Some of these examples highlight advantages of imaging flow cytometry for certain research objectives, but we also include examples that would not necessarily require imaging and could be performed on a conventional cytometer to demonstrate other concepts in cytometric evaluation of microalgae. We demonstrate the value of these approaches for (1) analysis of populations, (2) documentation of cellular features, and (3) analysis of gene expression.

Photobiological hydrogen production is of great importance because of its promise for generating clean renewable energy. In nature, green algae cannot produce hydrogen as a result of the extreme sensitivity of hydrogenase to oxygen. However, we find that silicification-induced green algae aggregates can achieve sustainable photobiological hydrogen production even under natural aerobic conditions. The core-shell structure of the green algae aggregates creates a balance between photosynthetic electron generation and hydrogenase activity, thus allowing the production of hydrogen. This finding provides a viable pathway for the solar-driven splitting of water into hydrogen and oxygen to develop green energy alternatives by using rationally designed cell-material complexes., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study investigated the feasibility of lipid production of Chlorella sp. from waste materials. Lipid-extracted microalgal biomass residues (LMBRs) and molasses were hydrolyzed, and their hydrolysates were analyzed. Five different hydrolysate mixture ratios (w/w) of LMBRs/molasses (1/0, 1/1, 1/4, 1/9, and 0/1) were used to cultivate Chlorella sp. The results showed that carbohydrate and protein were the two main compounds in the LMBRs, and carbohydrate was the main compound in the molasses. The highest biomass concentration of 5.58 g/L, Y biomass/sugars of 0.59 g/g, lipid productivity of 335 mg/L/day, and Y lipids/sugars of 0.25 g/g were obtained at the hydrolysate mixture ratio of LMBRs/molasses of 1/4. High C/N ratio promoted the conversion of sugars into lipids. The lipids extracted from Chlorella sp. shared similar lipid profile of soybean oil and is therefore a potential viable biodiesel feedstock. These results showed that Chlorella sp. can utilize mixed sugars and amino acids from LMBRs and molasses to accumulate lipids efficiently, thus reducing the cost of microalgal biodiesel production and improving its economic viability.

Exploitation of magnetic flocculants is regarded as a very promising energy-saving approach to microalgae harvesting. However, its practical applicability remains limited, mainly because of the problem of the postharvest separation of magnetic flocculants from microalgal flocs, which is crucial both for magnetic-flocculant recycling and high-purity microalgal biomasses, but which is also a very challenging and energy-consuming step. In the present study, we designed magnetic nanoflocculants dually functionalizable by two different organosilane compounds, (3-aminopropyl)triethoxysilane (APTES) and octyltriethoxysilane (OTES), which flocculate negatively charged microalgae and are readily detachable at the water-nonpolar organic solvent (NOS) interface only by application of an external magnetic field. APTES functionalization imparts a positive zeta potential charge (29.6 mV) to magnetic nanoflocculants, thereby enabling microalgae flocculation with 98.5% harvesting efficiency (with a dosage of 1.6 g of dMNF/g of cells). OTES functionalization imparts lipophilicity to magnetic nanoflocculants to make them compatible with NOS, thus effecting efficient separation of magnetic flocculants passing through the water-NOS interface sieve from hydrophilic microalgae. Our new energy-saving approach to microalgae harvesting concentrates microalgal cultures (∼1.5 g/L) up to 60 g/L, which can be directly connected to the following process of NOS-assisted wet lipid extraction or biodiesel production, and therefore provides, by simplifying multiple downstream processes, a great potential cost reduction in microalgae-based biorefinement.

In this study, a numerical simulation using computational fluid dynamics (CFD) was used to investigate the hydrodynamic characteristics of circular ponds with three different impellers (hydrofoil, four-pitched-blade turbine, and grid plate). The reliability of the CFD model was validated by particle image velocimetry (PIV). Hydrodynamic analyses were conducted to evaluate the average velocity magnitude along the light direction (Uz), turbulence properties, average shear stress, pressure loss and the volume percentage of dead zone inside circular ponds. The simulation results showed that Uz value of hydrofoil was 58.9, 40.3, and 28.8% higher than those of grid plate with single arm, grid plate with double arms and four-pitched blade turbines in small-scale circular ponds, respectively. In addition, hydrofoil impeller with down-flow operation had outstanding mixing characteristics. Lastly, the results of Chlorella pyrenoidosa cultivation experiments indicated that the biomass concentration of hydrofoil impeller with down-flow operation was 65.2 and 88.8% higher than those of grid plate with double arms and four-pitched-blade turbine, respectively. Therefore, the optimal circular pond mixing system for microalgae cultivation involved a hydrofoil impeller with down-flow operation.

Sulfur deficiency in plant cells has not been considered as a potential abiotic factor that can induce oxidative stress. We studied the antioxidant defense system of Chlorella sorokiniana cultured under sulfur (S) deficiency, imposed for a maximum period of 24 h, to evaluate the effect of an S shortage on oxidative stress. S deprivation induced an immediate (30 min) but transient increase in the intracellular H2O2 content, which suggests that S limitation can lead to a temporary redox disturbance. After 24 h, S deficiency in Chlorella cells decreased the glutathione content to <10% of the value measured in cells that were not subjected to S deprivation. Consequently, we assumed that the cellular antioxidative mechanisms could be altered by a decrease in the total glutathione content. The total ascorbate pool increased within 2 h after the initiation of S depletion, and remained high until 6 h; however, ascorbate regeneration was inhibited under limited S conditions, indicated by a significant decrease in the ascorbate/dehydroascorbate (AsA/DHA) ratios. Furthermore, ascorbate peroxidase (APX) and superoxide dismutase (SOD) were activated under S deficiency, but we assumed that these enzymes were involved in maintaining the cellular H2O2 balance for at least 4 h after the initiation of S starvation. We concluded that S deprivation triggers redox changes and induces antioxidant enzyme activities in Chlorella cells. The accumulation of total ascorbate, changes in the reduced glutathione/oxidized glutathione (GSH/GSSG) ratios and an increase in the activity of SOD and APX enzymes indicate that oxidative perturbation occurs during S deprivation., (© The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Industrial wastewaters often contain high levels of metal mixtures, in which metal mixtures may have synergistic or antagonistic effects on aquatic organisms. A combination of metallomics and nuclear magnetic resonance spectroscopy (NMR)-based metabolomics was employed to understand the consequences of multi-metal systems (Cu, Cd, Pb) on freshwater microalgae. Morphological characterization, cell viability and chlorophyll a determination of metal-spiked Chlorella sp. suggested synergistic effects of Cu and Cd on growth inhibition and toxicity. While Pb has no apparent effect on Chlorella sp. metabolome, a substantial decrease of sucrose, amino acid content and glycerophospholipid precursors in Cu-spiked microalgae revealed Cu-induced oxidative stress. Addition of Cd to Cu-spiked cultures induced more drastic metabolic perturbations, hence we confirmed that Cu and Cd synergistically influenced photosynthesis inhibition, oxidative stress and membrane degradation. Total elemental analysis revealed a significant decrease in K, and an increase in Na, Mg, Zn and Mn concentrations in Cu-spiked cultures. This indicated that Cu is more toxic to Chlorella sp. as compared to Cd or Pb, and the combination of Cu and Cd has a strong synergistic effect on Chlorella sp. oxidative stress induction. Oxidative stress is confirmed by liquid chromatography tandem mass spectrometry analysis, which demonstrated a drastic decrease in the GSH/GSSG ratio solely in Cu-spiked cultures. Interestingly, we observed Cu-facilitated Cd and Pb bioconcentration in Chlorella sp. The absence of phytochelatins and an increment of extracellular polymeric substances (EPS) yields in Cu-spiked cultures suggested that the mode of bioconcentration of Cd and Pb is through adsorption of free metals onto the algal EPS rather than intracellular chelation to phytochelatins.

Integrated and genome-based flux balance analysis, metabolomics, and (13)C-label profiling of phototrophic and heterotrophic metabolism in Chlorella protothecoides, an oleaginous green alga for biofuel. The green alga Chlorella protothecoides, capable of autotrophic and heterotrophic growth with rapid lipid synthesis, is a promising candidate for biofuel production. Based on the newly available genome knowledge of the alga, we reconstructed the compartmentalized metabolic network consisting of 272 metabolic reactions, 270 enzymes, and 461 encoding genes and simulated the growth in different cultivation conditions with flux balance analysis. Phenotype-phase plane analysis shows conditions achieving theoretical maximum of the biomass and corresponding fatty acid-producing rate for phototrophic cells (the ratio of photon uptake rate to CO2 uptake rate equals 8.4) and heterotrophic ones (the glucose uptake rate to O2 consumption rate reaches 2.4), respectively. Isotope-assisted liquid chromatography-mass spectrometry/mass spectrometry reveals higher metabolite concentrations in the glycolytic pathway and the tricarboxylic acid cycle in heterotrophic cells compared with autotrophic cells. We also observed enhanced levels of ATP, nicotinamide adenine dinucleotide (phosphate), reduced, acetyl-Coenzyme A, and malonyl-Coenzyme A in heterotrophic cells consistently, consistent with a strong activity of lipid synthesis. To profile the flux map in experimental conditions, we applied nonstationary (13)C metabolic flux analysis as a complementing strategy to flux balance analysis. The result reveals negligible photorespiratory fluxes and a metabolically low active tricarboxylic acid cycle in phototrophic C. protothecoides. In comparison, high throughput of amphibolic reactions and the tricarboxylic acid cycle with no glyoxylate shunt activities were measured for heterotrophic cells. Taken together, the metabolic network modeling assisted by experimental metabolomics and (13)C labeling better our understanding on global metabolism of oleaginous alga, paving the way to the systematic engineering of the microalga for biofuel production., (© 2015 American Society of Plant Biologists. All Rights Reserved.)

Chlorella, the archetype of unicellular green algae, is a high-performance primary producer in aquatic and terrestrial ecosystems. Under the simple spherical morphology of Chlorella, many other 'green balls' unfolded as independent phylogenetic lineages as a result of convergent evolution. By contrast, green algae with strikingly different phenotypes were unmasked as close relatives of Chlorella by modern molecular techniques. Here, we point to the increasing impact of these diverse protists on ecology, evolution, and biotechnology in the light of integrative taxonomy., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The aim of this study was to investigate the effects of various organic carbon sources (glucose, galactose, fructose, sucrose, maltose, lactose and starch) on the growth and biochemical composition of Chlorella pyrenoidosa. Monosaccharides were found to exert stronger stimulative effects on the algal growth than disaccharides and starch. After 10-day culture, addition of 0.5-5.0 g L(-1) glucose and galactose significantly reduced the cellular protein contents by 27.7-63.7% and 22.6-60.5%, respectively, and significantly increased the carbohydrate contents by 103.2-266.5% and 91.9-240.0%, respectively. However, addition of 0.5-5.0 g L(-1) disaccharides and starch did not significantly affect the contents of lipid, protein and carbohydrate. Similar to the normal nitrogen condition, the cellular biochemical composition was not significantly affected by addition of 3.0 g L(-1) disaccharides and starch under the low nitrogen condition. Finally, the significance of this work in the biotechnological application of mixotrophic cultivation of C. pyrenoidosa was further discussed., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Though less attention has been paid to microalgae as a feedstock for bioethanol production, many microalgae seem to have this potential since they contain no lignin, minor hemicellulose, and abundant carbohydrate. The objective of this study was to investigate the effect of nitrogen starvation on carbohydrate and starch accumulation in green microalga Chlorella zofingiensis and assess the feasibility of using this microalga as a bioethanol feedstock. The results showed that the specific growth rate under nitrogen starvation (0.48 day(-1)) was much lower than that under nitrogen repletion (1.02 day(-1)). However, nitrogen starvation quickly induced the accumulation of carbohydrate, especially starch. After merely 1 day of nitrogen starvation, carbohydrate and starch increased 37 % and 4.7-fold, respectively. The highest carbohydrate content reached 66.9 % of dry weight (DW), and 66.7 % of this was starch. In order to obtain enough carbohydrate productivities for bioethanol production, two-stage cultivation strategy was implemented and found to be effective for enhancing biomass, carbohydrate, and starch simultaneously. The optimal biomass, carbohydrate, and starch productivities of C. zofingiensis were obtained after 5 days of cultivation, and their values were 699, 407, and 268 mg L(-1) day(-1), respectively.

In this study, Chlorella zofingiensis harvesting by dissolved air flotation (DAF) was critically evaluated with regard to algal concentration, culture conditions, type and dosage of coagulants, and recycle ratio. Harvesting efficiency increased with coagulant dosage and leveled off at 81%, 86%, 91%, and 87% when chitosan, Al(3+) , Fe(3+) , and cetyl trimethylammonium bromide (CTAB) were used at dosages of 70, 180, 250, and 500 mg g(-1) , respectively. The DAF efficiency-coagulant dosage relationship changed with algal culture conditions. Evaluation of the influence of the initial algal concentration and recycle ratio revealed that, under conditions typical for algal harvesting, it is possible that the number of bubbles is insufficient. A DAF algal harvesting model was developed to explain this observation by introducing mass-based floc size distributions and a bubble limitation into the white water blanket model. The model revealed the importance of coagulation to increase floc-bubble collision and attachment, and the preferential interaction of bubbles with larger flocs, which limited the availability of bubbles to the smaller sized flocs. The harvesting efficiencies predicted by the model agree reasonably with experimental data obtained at different Al(3+) dosages, algal concentrations, and recycle ratios. Based on this modeling, critical parameters for efficient algal harvesting were identified., (© 2014 Wiley Periodicals, Inc.)

Electrochemical characteristics of O2 and/or mediators secreted by a single cell of Chlorella fusea were analyzed by using amperometric measurements on microelectrodes embedded in a microfluidic device. A single cell was trapped in a microfluidic channel, which simplifies the mass transfer phenomenon, i.e., one-dimensional distribution of solutes in the channel. Such amperometric measurements allowed us to obtain more refined data in a localized space and to understand photosynthetic behavior of algae at the single cell level. In addition, the concentration of a photosynthetic mediator, p-benzoquinone, was numerically calculated by using the finite element method.

The present study investigates the effects of different physico-chemical parameters for the growth of Chlorella sorokiniana and subsequently determination of nutritional values of its biomass. Most suitable temperature, light intensity, pH, and acetic acid concentration were 30°C, 100 μmol m(-2)s(-1), pH 7.5, and 34.8mM, respectively for the growth of this microorganism. Arrhenius growth activation energy, Ea was calculated as 7.08 kJ mol(-1). Monod kinetics constants: maximum specific growth rate (μ max) and substrate (acetic acid) affinity coefficient (Ks) were determined as 0.1 ± 0.01 h(-1) and 76 ± 8 mg L(-1), respectively. Stoichiometric analysis revealed the capture of 1.83 g CO2 and release of 1.9 g O2 for 1g algal biomass synthesis. Algal biomass of C. sorokiniana was found rich in protein and several important minerals such as Mg, Ca, and Fe. Astaxanthin and β-carotene were extracted and quantified using high performance liquid chromatography., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Self-regulation of temperature in microchip systems is crucial for their applications in biomedical fields such as cell culture and biomolecule synthesis as well as those cases that require constant temperature conditions. Here we report on a simple and versatile approach for in situ fabrication of a smart hydrogel microvalve within a microchip for thermostatic control. The thermo-responsive hydrogel microvalve enables the "on-off" switch by sensing temperature fluctuations to control the fluid flux as well as the fluid heat exchange for self-regulation of the temperature at a constant range. Such temperature self-regulation is demonstrated by integrating the microvalve-incorporated microchip into the flow circulation loop of a micro-heat-exchanging system for thermostatic control. Moreover, the microvalve-incorporated microchip is employed for culturing cells under temperature self-regulation. The smart microvalve shows great potential as a temperature controller for applications that require thermostatic conditions. This approach offers a facile and flexible strategy for in situ fabricating hydrogel microvalves within microchips as chemostats and microreactors for biomedical applications.

This article presents a study on screening of microalgal strains from the Peking University Algae Collection and heterotrophic cultivation for biodiesel production of a selected microalgal strain. Among 89 strains, only five were capable of growing under heterotrophic conditions in liquid cultures and Chlorella sp. PKUAC 102 was found the best for the production of heterotrophic algal biodiesel. Composition of the growth medium was optimised using response surface methodology and optimised growth conditions were successfully used for cultivation of the strain in a fermentor. Conversion of algal lipids to fatty acid methyl esters (FAMEs) showed that the lipid profile of the heterotrophically cultivated Chlorella sp. PKUAC 102 contains fatty acids suitable for biodiesel production.

To determine the optimum light intensity per cell required for rapid growth regardless of cell density, continuous cultures of the microalga Chlorella zofingiensis were grown with a sufficient supply of nutrients and CO2 and were subjected to different light intensities in the range of 75-1000 μE m(-2) s(-1). The cell density of culture increased over time for all light conditions except for the early stage of the high light condition of 1000 μE m(-2) s(-1). The light intensity per cell required for the high specific growth rate of 0.5 day(-1) was determined to be 28-45 μE g-ds(-1) s(-1). The specific growth rate was significantly correlated to light intensity (y=0.721×x/(66.98+x), r(2)=0.85, p<0.05). A high specific growth rate was maintained over a range of light intensities (250-1000 μE m(-2) s(-1)). This range of light intensities suggested that effective production of C. zofingiensis can be maintained outdoors under strong light by using the optimum specific light intensity., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The present study reports evaluation of an indigenous microalgal isolate Chlorella sp. FC2 IITG as a potential candidate for biodiesel production. Characterization of the strain was performed under photoautotrophic, heterotrophic, and mixotrophic cultivation conditions. Further, an open-pond cultivation of the strain under outdoor conditions was demonstrated to evaluate growth performance and lipid productivity under fluctuating environmental parameters and in the presence of potential contaminants. The key findings were: (1) the difference in cultivation conditions resulted in significant variation in the biomass productivity (73-114 mg l⁻¹ day⁻¹) and total lipid productivity (35.02-50.42 mg l⁻¹ day⁻¹) of the strain; (2) nitrate and phosphate starvation were found to be the triggers for lipid accumulation in the cell mass; (3) open-pond cultivation of the strain under outdoor conditions resulted in biomass productivity of 44 mg l⁻¹ day⁻¹ and total lipid productivity of 10.7 mg l⁻¹ day⁻¹; (4) a maximum detectable bacterial contamination of 7 % of the total number of cells was recorded in an open-pond system; and (5) fatty acid profiling revealed abundance of palmitic acid (C16:0), oleic acid (C18:1) and linoleic acid (C18:2), which are considered to be the key elements for suitable quality biodiesel.

The aim of this work was to study the effect of Se(+VI) on viability, cell morphology, and selenomethionine accumulation of the green alga Chlorella sorokiniana grown in batch cultures. Culture exposed to sublethal Se concentrations of 40 mg · L(-1) (212 μM) decreased growth rates for about 25% compared to control. A selenate EC50 value of 45 mg · L(-1) (238.2 μM) was determined. Results showed that chlorophyll and carotenoids contents were not affected by Se exposure, while oxygen evolution decreased by half. Ultrastructural studies revealed granular stroma, fingerprint-like appearance of thylakoids which did not compromise cell activity. Unlike control cultures, SDS PAGE electrophoresis of crude extracts from selenate-exposed cell cultures revealed appearance of a protein band identified as 53 kDa Rubisco large subunit of Chlorella sorokiniana, suggesting that selenate affects expression of the corresponding chloroplast gene as this subunit is encoded in the chloroplast DNA. Results revealed that the microalga was able to accumulate up to 140 mg · kg(-1) of SeMet in 120 h of cultivation. This paper shows that Chlorella sorokiniana biomass can be enriched in the high value aminoacid SeMet in batch cultures, while keeping photochemical viability and carbon dioxide fixation activity intact, if exposed to suitable sublethal concentrations of Se.

We recently isolated a small green alga from a biological sand crust (BSC) in the NW Negev, Israel. Based on its 18S rRNA and rbcL genes, it is a close relative of Chlorella sorokiniana and of certain strains of C. vulgaris and C. variabilis, but differs substantially in many aspects from C. sorokiniana. Because the classification of Chlorellales is still not resolved, we designated this species as C. ohadii (Trebouxiophyceae) in honor of Professor Itzhak Ohad. Under controlled laboratory conditions, C. ohadii showed marked structural and photosynthetic performance changes, depending on the carbon source used during growth, as well as remarkable resistance to photoinhibition. CO2 -dependent O2 evolution was not affected even when exposed to a light intensity of 3500 μmole photons m(-2)  s(-1) , over 1.5 times the maximal intensity reached at the BSC surface, whereas the variable fluorescence declined sharply. We briefly discuss the use of fluorescence to assess photosynthetic rate and the implications of this finding for the assessment of global BSCs activity., (© 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)

To extract lipids from wet microalgae through cell disruption, the effects of microwave treatment on the dynamic cell wall microstructures were investigated. The fractal dimension of raw, untreated microalgal cells was 1.46. The disruption level of microalgal cell walls was enhanced when microwave treatment temperature increased from 80 to 120°C, resulting in an increase in microalgal cell fractal dimension from 1.61 to 1.91. The cell wall thickness and pore diameters in cell walls increased from 0.11 to 0.59 μm and from 0.005 to 0.18 μm, respectively, when microwave treatment time increased from 0 to 20 min. The outer pectin layers of cell walls gradually detached and the porosity of inner cellulose layers increased when microwave treatment time increased to 26 min. The initial point of disruption appeared at the maximum curvature (approximately 1.01×10(7) m(-1)) of cell walls. Numbers of short-chain and saturated lipids increased because of microwave electromagnetic effect., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

This work aimed to develop a rotating algal biofilm (RAB) cultivation system that can be widely adopted by microalgae producers for easy biomass harvest. Algal cells were grown on the surface of a material rotating between nutrient-rich liquid and CO2-rich gaseous phase. Scrapping biomass from the attached surface avoided the expensive harvest operations such as centrifugation. Among various attachment materials, cotton sheet resulted in best algal growth, durability, and cost effectiveness. A lab-scale RAB system was further optimized with harvest frequency, rotation speed, and CO2 levels. The algal biomass from the RAB system had a similar water content as that in centrifuged biomass. An open pond raceway retrofitted with a pilot-scale RAB system resulted in a much higher biomass productivity when compared to a control open pond. Collectively, the research shows that the RAB system is an efficient algal culture system for easy biomass harvest with enhanced biomass productivity., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Microalgae are one of the most promising sustainable energy sources for biodiesel production. However, the high costs of the downstream process are a major bottleneck for commercial-scale production of biodiesel from a microalgal biomass. A novel approach called the cationic surfactant-based harvesting and cell disruption (CSHD) method was studied to determine its effectiveness in simultaneous microalgal biomass harvesting and cell disruption. Using CSHD, the harvesting efficiency reached more than 91% in less than 5 min and 97% in 90 min. Moreover, CSHD exhibited a powerful ability to disrupt the cells; the lipid recovery was increased 133% compared to not using CSHD. CSHD allowed the extraction of up to 100% of the total lipids from a wet microalgal biomass with 80% water content. All of these results were achieved without using energy-intensive equipment. Altogether, our results suggest that CSHD is an energy-efficient technique for the downstream process of microalgal lipid production., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Hard-shell case: Using a (RKK)4 D8 peptide allows mineralization to occur under cytocompatible conditions. Thus individual Chlorella cells could be encapsulated within a SiO2 -TiO2 nanoshell with high cell viability (87 %). The encapsulated Chlorella showed an almost threefold increase in their thermo-tolerance after 2 h at 45 °C., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This work for the first time investigated lipid accumulation by a two-stage regime namely heterotrophic-Cu(II) stressed (HCuS) and underlying molecular basis of lipid biosynthesis in Chlorella protothecoides cells. The results showed that the optimized biomass and lipid yield were achieved by 6.47 g/L and 5.78 g/L with this strategy. The fatty acids compositions (almost 100% of them are C15 to C20) are ideal for preparing high quality biodiesel. Further, 30 differentially expressed proteins response to HCuS were involved in carbohydrate metabolism, carbon fixation, TCA cycle, lipid metabolism, protein biosynthesis, transportation and regulation, ATP and RNA biosynthesis, nucleotide metabolism, ROS scavenging. Especially, glycolysis pathway might be the important contributor for lipid accumulation. In future, further functional analysis of these altered proteins would help to reveal more concerning lipid biosynthesis pathway., (Copyright © 2013 Elsevier Ltd. All rights reserved.)