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Microalgae show great promise for producing valuable molecules like biofuels, but their large-scale production faces challenges, with harvesting being particularly expensive due to their low concentration in water, necessitating extensive treatment. While methods such as centrifugation and filtration have been proposed, their efficiency and cost-effectiveness are limited. Flotation, involving air-bubbles lifting microalgae to the surface, offers a viable alternative, yet the repulsive interaction between bubbles and cells can hinder its effectiveness. Previous research from our group proposed using an amphiphilic chitosan derivative, polyoctyl chitosan (PO-chitosan), to functionalize bubbles used in dissolved air flotation (DAF). Molecular-scale studies performed using atomic force microscopy (AFM) revealed that PO-chitosan's efficiency correlates with cell surface properties, particularly hydrophobic ones, raising the question of whether this molecule can in fact be used more generally to harvest different microalgae. Evaluating this, we used a different strain of Chlorella vulgaris and first characterized its surface properties using AFM. Results showed that cells were hydrophilic but could still interact with PO-chitosan on bubble surfaces through a different mechanism based on specific interactions. Although force levels were low, flotation resulted in 84% separation, which could be explained by the presence of AOM (algal organic matter) that also interacts with functionalized bubbles, enhancing the overall separation. Finally, flocculation was also shown to be efficient and pH-independent, demonstrating the potential of PO-chitosan for harvesting microalgae with different cell surface properties and thus for further sustainable large-scale applications.

The composition of species and the physiological status of microalgal cells serve as significant indicators for monitoring marine environments. Symbiotic with corals, Symbiodiniaceae are more sensitive to the environmental response. However, current methods for evaluating microalgae tend to be population-based indicators that cannot be focused on single-cell level, ignoring potentially heterogeneous cells as well as cell state transitions. In this study, we proposed a microalgal cell detection method based on computer vision and microfluidics, which combined microscopic image processing, microfluidic chip and convolutional neural network to achieve label-free, sheathless, automated and high-throughput microalgae identification and cell state assessment. By optimizing the data import, training process and model architecture, we solved the problem of identifying tiny objects at the micron scale, and the optimized model was able to perform the tasks of cell multi-classification and physiological state assessment with more than 95% mean average precision. We discovered a novel transition state and explored the thermal sensitivity of three clades of Symbiodiniaceae, and discovered the phenomenon of cellular heat shock at high temperatures. The evolution of the physiological state of Symbiodiniaceae cells is very important for directional cell evolution and early warning of coral ecosystem health.

Macroalgae are multicellular, aquatic autotrophs that play vital roles in global climate maintenance and have diverse applications in biotechnology and eco-engineering, which are directly linked to their multicellularity phenotypes. However, their genomic diversity and the evolutionary mechanisms underlying multicellularity in these organisms remain uncharacterized. In this study, we sequenced 110 macroalgal genomes from diverse climates and phyla, and identified key genomic features that distinguish them from their microalgal relatives. Genes for cell adhesion, extracellular matrix formation, cell polarity, transport, and cell differentiation distinguish macroalgae from microalgae across all three major phyla, constituting conserved and unique gene sets supporting multicellular processes. Adhesome genes show phylum- and climate-specific expansions that may facilitate niche adaptation. Collectively, our study reveals genetic determinants of convergent and divergent evolutionary trajectories that have shaped morphological diversity in macroalgae and provides genome-wide frameworks to understand photosynthetic multicellular evolution in aquatic environments., (Copyright © 2024 The Author. Published by Elsevier Inc. All rights reserved.)

Cell attachment onto electrode-forming biocathodes is a promising alternative to expensive catalysts used for electricity production in bioelectrochemical systems (BESs). Though BESs have been extensively studied for decades, the processes, underlying mechanisms, and determinant driving forces of microalgal biocathode formation remain largely unknown. In this study, we employed a model unicellular motile microalga, Chlamydomonas microsphaera, to investigate the microalgal attachment processes onto the electrode surface of a BES and to identify the determinant factors. Results showed that the initial attachment of C. micrrosphaera cells is determined by the applied external voltage rather than nutrient availability and occurs via electrotaxis-mediated cell motility. The subsequent development of the C. microsphaera biofilm is then increasingly determined by nutrient availability. Our results revealed that, in the absence of an external voltage, nutrient availability remains a dominant factor controlling the fate of the microalgal surface attachment and subsequent biofilm formation processes. Thus, our results show that electrotactic and chemotactic movements are crucial to facilitate the initial attachment and subsequent biofilm formation of C. microsphaera onto the electrode surfaces of BES. This study provides new insights into the mechanisms of microalgal surface attachment and biofilm formation processes on microalgal biocathodes, which hold great promise for improving the electrochemical properties of cathodes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Microalgae are photosynthetic organisms widely distributed in nature and serve as a sustainable source of bioproducts. Their carbohydrate components are also promising candidates for bioenergy production and bioremediation, but the structural characterization of these heterogeneous polymers in cells remains a formidable problem. Here we present a widely applicable protocol for identifying and quantifying the glycan content using magic-angle-spinning (MAS) solid-state NMR (ssNMR) spectroscopy, with validation from glycosyl linkage and composition analysis deduced from mass-spectrometry (MS). Two-dimensional 13 C- 13 C correlation ssNMR spectra of a uniformly 13 C-labeled green microalga Parachlorella beijerinckii reveal that starch is the most abundant polysaccharide in a naturally cellulose-deficient strain, and this polymer adopts a well-organized and highly rigid structure in the cell. Some xyloses are present in both the mobile and rigid domains of the cell wall, with their chemical shifts partially aligned with the flat-ribbon 2-fold xylan identified in plants. Surprisingly, most other carbohydrates are largely mobile, regardless of their distribution in glycolipids or cell walls. These structural insights correlate with the high digestibility of this cellulose-deficient strain, and the in-cell ssNMR methods will facilitate the investigations of other economically important algae species.

Imbalanced light absorption by photosystem I (PSI) and photosystem II (PSII) in oxygenic phototrophs leads to changes in interaction of photosystems altering the linear electron flow. In plants and green algae, this imbalance is mitigated by a partial migration of the chlorophyll a/b containing light-harvesting antenna between the two photosystem core complexes. This migration is registered as fluorescence changes of the pigment apparatus and is termed the reverse transitions between States 1 and 2. By contrast, the molecular mechanism of State 1/2 transitions in phycobilisome (PBS)-containing photosynthetics, cyanobacteria and red algae, is still insufficiently understood. The suggested hypotheses - PBS movement along the surface of thylakoid membrane between PSI and PSII complexes, reversible PBS detachment from the dimeric PSII complex, and spillover - have some limitations as they do not fully explain the accumulated data. Here, we have recorded changes in the stationary fluorescence emission spectra of red algae and cyanobacteria in States 1/2 at room temperature, which allowed us to offer an explanation of the existing contradictions. The change of room temperature fluorescence of chlorophyll belonged to PSII was revealed, while the fluorescence of PBS associated with the PSII complexes remained during States 1/2 transitions at the stable level. Only the reversible dissociation of PBS from the monomeric PSI was revealed earlier which implied different degree of surface contact of PBS with the two photosystems. The detachment of PBS from the PSI corresponds to ferredoxin oxidation as electron carrier and the increase of cyclic electron transport in the pigment apparatus in State I.

Microalgae offer a promising source of biofuel and a wide array of high-value biomolecules. Large-scale cultivation of microalgae at low density poses a significant challenge in terms of water management. High-density microalgae cultivation, however, can be challenging due to biochemical changes associated with growth dynamics. Therefore, there is a need for a biomarker that can predict the optimum density for high biomass cultivation. A locally isolated microalga Cyanobacterium aponinum CCC734 was grown with optimized nitrogen and phosphorus in the ratio of 12:1 for sustained high biomass productivity. To understand density-associated bottlenecks secretome dynamics were monitored at biomass densities from 0.6 ± 0.1 to 7 ± 0.1 g/L (2 to 22 OD) in batch mode. Liquid chromatography coupled with mass spectrometry identified 880 exometabolites in the supernatant of C. aponinum CCC734. The PCA analysis showed similarity between exometabolite profiles at low (4 and 8 OD) and mid (12 and 16 OD), whereas distinctly separate at high biomass concentrations (20 and 22 OD). Ten exometabolites were selected based on their role in influencing growth and are specifically present at low, mid, and high biomass concentrations. Taking cues from secretome dynamics, 5.0 ± 0.5 g/L biomass concentration (16 OD) was optimal for C. aponinum CCC734 cultivation. Further validation was performed with a semi-turbidostat mode of cultivation for 29 days with a volumetric productivity of 1.0 ± 0.2 g/L/day. The secretomes-based footprinting tool is the first comprehensive growth study of exometabolite at the molecular level at variable biomass densities. This tool may be utilized in analyzing and directing microalgal cultivation strategies and reduction in overall operating costs., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Endosymbioses have shaped the evolutionary trajectory of life and remain ecologically important. Investigating oceanic photosymbioses can illuminate how algal endosymbionts are energetically exploited by their heterotrophic hosts and inform on putative initial steps of plastid acquisition in eukaryotes. By combining three-dimensional subcellular imaging with photophysiology, carbon flux imaging, and transcriptomics, we show that cell division of endosymbionts ( Phaeocystis ) is blocked within hosts (Acantharia) and that their cellular architecture and bioenergetic machinery are radically altered. Transcriptional evidence indicates that a nutrient-independent mechanism prevents symbiont cell division and decouples nuclear and plastid division. As endosymbiont plastids proliferate, the volume of the photosynthetic machinery volume increases 100-fold in correlation with the expansion of a reticular mitochondrial network in close proximity to plastids. Photosynthetic efficiency tends to increase with cell size, and photon propagation modeling indicates that the networked mitochondrial architecture enhances light capture. This is accompanied by 150-fold higher carbon uptake and up-regulation of genes involved in photosynthesis and carbon fixation, which, in conjunction with a ca.15-fold size increase of pyrenoids demonstrates enhanced primary production in symbiosis. Mass spectrometry imaging revealed major carbon allocation to plastids and transfer to the host cell. As in most photosymbioses, microalgae are contained within a host phagosome (symbiosome), but here, the phagosome invaginates into enlarged microalgal cells, perhaps to optimize metabolic exchange. This observation adds evidence that the algal metamorphosis is irreversible. Hosts, therefore, trigger and benefit from major bioenergetic remodeling of symbiotic microalgae with potential consequences for the oceanic carbon cycle. Unlike other photosymbioses, this interaction represents a so-called cytoklepty, which is a putative initial step toward plastid acquisition., Competing Interests: The authors declare no competing interest.

Since the removal of contaminations in microalgal cultures is extremely laborious and time-consuming, we developed a rapid workflow to obtain axenicity by a combination of fluorescence-activated cell sorting (FACS) and plate spreading. During method development, several cyanobacteria and green algae strains were successfully made axenic. At the end, method transferability to another FACS device was demonstrated. Our workflow offers great time-savings with less hands-on laboratory work compared to conventional isolation techniques., (Copyright © 2021. Published by Elsevier B.V.)

The extremophilic unicellular red microalga Galdieria sulphuraria (Cyanidiophyceae) is able to grow autotrophically, or mixo- and heterotrophically with 1% glycerol as a carbon source. The alga divides by multiple fission into more than two cells within one cell cycle. The optimal conditions of light, temperature and pH (500 µmol photons m -2 s -1 , 40 °C, and pH 3; respectively) for the strain Galdieria sulphuraria (Galdieri) Merola 002 were determined as a basis for synchronization experiments. For synchronization, the specific light/dark cycle, 16/8 h was identified as the precondition for investigating the cell cycle. The alga was successfully synchronized and the cell cycle was evaluated. G. sulphuraria attained two commitment points with midpoints at 10 and 13 h of the cell cycle, leading to two nuclear divisions, followed subsequently by division into four daughter cells. The daughter cells stayed in the mother cell wall until the beginning of the next light phase, when they were released. Accumulation of glycogen throughout the cell cycle was also described. The findings presented here bring a new contribution to our general understanding of the cell cycle in cyanidialean red algae, and specifically of the biotechnologically important species G. sulphuraria .

Microalgae are versatile microorganisms with applications in food, energy and fine chemicals, among others. Modelling the dynamics of microalgae inside a photobioreactor is a convenient and inexpensive way to determine the concentration of cells over time. Numerous models have been developed in the literature, but only a few are able to give relevant biological information. Such information can then be used to further improve the production process. The objective of this work was to develop a model for the determination of microalgal dynamics inside a photobioreactor as a function of the environmental conditions, to retrieve the size-specific growth and division rates as well as the number of daughter cells. The results demonstrate how to evaluate the time needed for microalgae to complete a full life-cycle. Inexpensive laboratory-based procedures and mathematical modelling are combined for the determination of relevant biological parameters., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Controlled nitrate feeding strategies for fed-batch cultures of microalgae were applied for the enhancement of lipid production and microalgal growth rates. In particular, in this study, the effect of nitrate feeding rates on lipid and biomass productivities in fed-batch cultures of Nannochloropsis gaditana were investigated using three feeding modes (i.e., pulse, continuous, and staged) and under two light variations on both lipid productivity and fatty acid compositions. Higher nitrate levels negatively affected lipid production in the study. Increasing the light intensity increased the lipid contents of the microalgae in all three fed-batch feeding modes. A maximum of 58.3% lipid- to dry weight ratio was achieved when using pulse-fed cultures at an illumination of 200 μmol photons m -2  s -1 and 10 mg/day of nitrate feeding. This condition also resulted in the maximum lipid productivity of 44.6 mg L -1  day -1 . The fatty acid compositions of the lipids consisted predominantly of long-chain fatty acids (C:16 and C:18) and accounted for 70% of the overall fatty acid methyl esters. Pulse feeding mode was found to significantly enhance the biomass and lipid production. The other two feeding modes (continuous and staged) were not ideal for lipid and biomass production. This study demonstrates the applicability of pulse feeding strategies in fed-batch cultures as an appropriate cultivation strategy that can increase both lipid accumulation and biomass production., (© 2021 American Institute of Chemical Engineers.)

Light attenuation is a primary challenge limiting the upscaling of photobioreactors for sustainable bio-production. One key to this challenge, is to model and optimise the light/dark cycles so that cells within the dark region can be frequently transferred to the light region for photosynthesis. Therefore, this study proposes the first mechanistic model to integrate the light/dark cycle effects into biomass growth kinetics. This model was initially constructed through theoretical derivation based on the intracellular reaction kinetics, and was subsequently modified by embedding a new parameter, effective light coefficient, to account for the effects of culture mixing. To generate in silico process data, a new multiscale reactive transport modelling strategy was developed to couple fluid dynamics with biomass growth kinetics and light transmission. By comparing against previous experimental and computational studies, the multiscale model shows to be of high accuracy. Based on its simulation result, an original correlation was proposed to link effective light coefficient with photobioreactor gas inflow rate; this has not been done before. The impact of this study is that by using the proposed mechanistic model and correlation, we can easily control and optimise photobioreactor gas inflow rates to alleviate light attenuation and maintain a high biomass growth rate., (© 2021 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals LLC.)

Microalgae are renewable, sustainable, and economical sources of biofuels and are capable of addressing pressing global demand for energy security. However, two challenging issues to produce high-level biofuels are to separate promising algal strains and protect biofuels from contamination of undesired bacteria, which rely on an economical and high-resolution separation technology. Separation technology based on induced-charge electroosmotic (ICEO) vortices offers excellent promise in economical microalga separation for producing biofuels because of its reconfigurable and flexible profiles and sensitive and precise selectivity. In this work, a practical ICEO vortex device is developed to facilitate high-resolution isolation of rich-lipid microalgae for the first time. We investigate electrokinetic equilibrium states of particles and particle-fluid ICEO effect in binary-particle manipulation. Nanoparticle separation is performed to demonstrate the feasibility and resolution of this device, yielding clear separation. Afterward, we leverage this technology in isolation of Chlorella vulgaris from heterogeneous microalgae with the purity exceeding 96.4%. Besides, this platform is successfully engineered for the extraction of single-cell Oocystis sp., obtaining the purity surpassing 95.2%. Moreover, with modulating parameters, we isolate desired-cell-number Oocystis sp. enabling us to investigate proliferation mode and carry out transcriptome analyses of Oocystis sp. for high-quality neutral lipids. This platform can be extended directly to economically separate other biological micro/nanosamples to address pressing issues, involving energy security, environmental monitoring, and disease diagnosis.

Melting of the Greenland Ice Sheet is a leading cause of land-ice mass loss and cryosphere-attributed sea level rise. Blooms of pigmented glacier ice algae lower ice albedo and accelerate surface melting in the ice sheet's southwest sector. Although glacier ice algae cause up to 13% of the surface melting in this region, the controls on bloom development remain poorly understood. Here we show a direct link between mineral phosphorus in surface ice and glacier ice algae biomass through the quantification of solid and fluid phase phosphorus reservoirs in surface habitats across the southwest ablation zone of the ice sheet. We demonstrate that nutrients from mineral dust likely drive glacier ice algal growth, and thereby identify mineral dust as a secondary control on ice sheet melting.

Bioprospecting for biodiesel potential in microalgae primarily involves a few model species of microalgae and rarely on non-model microalgae species. Therefore, the present study determined changes in physiology, oil accumulation, fatty acid composition and biodiesel properties of a non-model microalga Messastrum gracile SE-MC4 in response to 12 continuous days of nitrate-starve (NS) and nitrate-replete (NR) conditions respectively. Under NS, the highest oil content (57.9%) was achieved despite reductions in chlorophyll content, biomass productivity and lipid productivity. However, under both NS and NR, palmitic acid and oleic acid remained as dominant fatty acids thus suggesting high potential of M. gracile for biodiesel feedstock consideration. Biodiesel properties analysis returned high values of cetane number (CN 61.9-64.4) and degree of unsaturation (DU 45.3-57.4) in both treatments. The current findings show the possibility of a non-model microalga to inherit superior ability over model species in oil accumulation for biodiesel development.

Cryopreservation has been successfully used in the banking and maintenance of cultures of microorganisms, from bacteria to yeasts, since the onset of cryobiology. Biobanking of marine biological resources is crucial for development of scientific knowledge as researchers rely on guaranteed access to reliable, stable resources. Culture collections play a key role in the provision of marine biological resources as they ensure long-term ex situ storage of biological resources that are made available for public and private sector research and education. In this chapter, we provide protocols for cryopreservation of different types of algae cultures.

In this paper, we used a convolutional neural network to study the classification of marine microalgae by using low-resolution Mueller matrix images. Mueller matrix images of 12 species of algae from 5 families were measured by a Mueller matrix microscopy with an LED light source at 514 nm wavelength. The data sets of seven resolution levels were generated by the bicubic interpolation algorithm. We conducted two groups of classification experiments; one group classified the algae into 12 classes according to species category, and the other group classified the algae into 5 classes according to family category. In each group of classification experiments, we compared the classification results of the Mueller matrix images with those of the first element (M11) images. The classification accuracy of Mueller matrix images declines gently with the decrease of image resolution, while the accuracy of M11 images declines sharply. The classification accuracy of Mueller matrix images is higher than that of M11 images at each resolution level. At the lowest resolution level, the accuracy of 12-class classification and 5-class classification of full Mueller matrix images is 29.89% and 35.83% higher than those of M11 images, respectively. In addition, we also found that the polarization information of different species had different contributions to the classification. These results show that the polarization information can greatly improve the classification accuracy of low-resolution microalgal images.

A transmission hyperspectral microscopic imager (THMI) that utilizes machine learning algorithms for hyperspectral detection of microalgae is presented. The THMI system has excellent performance with spatial and spectral resolutions of 4 µm and 3 nm, respectively. We performed hyperspectral imaging (HSI) of three species of microalgae to verify their absorption characteristics. Transmission spectra were analyzed using principal component analysis (PCA) and peak ratio algorithms for dimensionality reduction and feature extraction, and a support vector machine (SVM) model was used for classification. The average accuracy, sensitivity and specificity to distinguish one species from the other two species were found to be 94.4%, 94.4% and 97.2%, respectively. A species identification experiment for a group of mixed microalgae in solution demonstrates the usability of the classification method. Using a random forest (RF) model, the growth stage in a phaeocystis growth cycle cultivated under laboratory conditions was predicted with an accuracy of 98.1%, indicating the feasibility to evaluate the growth state of microalgae through their transmission spectra. Experimental results show that the THMI system has the capability for classification, identification and growth stage estimation of microalgae, with strong potential for in-situ marine environmental monitoring and early warning detection applications.

A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.

Single-cell metabolite analysis plays an important role in biological study. While mass spectrometry is a powerful tool for identification and quantitation of metabolites, the low absolute analyte amounts in single cell and difficulty in sampling represent significant challenges in single cell analysis. In this study, we developed an effective method with a simple sampling procedure for analyzing single cells. A single cell was driven to a capillary tip through electro-migration, followed by releasing the cell contents through electroporation, into a sealed small volume (∼1.5 pL) to prevent dilution. Subsequent mass spectrometry analysis was performed directly with nanoelectrospray ionization. This method was applied for analyzing a variety of cells and monitoring the metabolic changes in response to perturbed cell culturing conditions. This method opens a new avenue for easy and rapid analysis of single cells with high sensitivity.

The unicellular photosynthetic organisms known as microalgae are becoming one of the most important models for aquatic system studies. Among them, Chlamydomonas reinhardtii is widely used as a bioindicator of pollution or of different changes in the environment. Numerous pollutants are present in aquatic environments, particularly plastics and nanoplastics. Physiological variations after an environmental change highlight variation in the macromolecular composition of microalgae (proteins, nucleic acids, lipids and carbohydrates). Recently, Fourier transform infrared vibrational spectroscopy has been described as a reliable tool, sensitive and allowing rapid measurement of macromolecular composition of microalgae. Coupled with preprocessing and principal component analysis, it is well adapted to monitoring the effect of environmental stress on biochemical composition. In this study, infrared spectroscopy, combined with multivariate analysis, has been tested first on known environmental stresses such as light intensity variation and nitrogen limitation. Then, this technique has been applied to monitor the interaction and potential impacts of polystyrene nanoparticles on microalgae. The results showed slight variations on protein and carbohydrates bands in the presence of nanoplastics, suggesting that their presence led to modifications in the biochemical composition of the microalgae. To confirm the interaction between microalgae and nanoplastics, visualization by confocal microscopy and cytotoxicity measurement has been carried out. Results showed that polystyrene nanoparticles seemed to adsorb on microalgae surface, leading to a loss of plasma membrane integrity. The resulting chemical modifications, even if moderate, could be detected by infrared spectroscopy' showing that this tool could be very helpful in the understanding of nanoparticle-microalgae interaction mechanisms.

We developed a low-cost multi-core inertial microfluidic centrifuge (IM-centrifuge) to achieve a continuous-flow cell/particle concentration at a throughput of up to 20 mL/min. To lower the cost of our IM-centrifuge, we clamped a disposable multilayer film-based inertial microfluidic (MFIM) chip with two reusable plastic housings. The key MFIM chip was fabricated in low-cost materials by stacking different polymer-film channel layers and double-sided tape. To increase processing throughput, multiplexing spiral inertial microfluidic channels were integrated within an all-in-one MFIM chip, and a novel sample distribution strategy was employed to equally distribute the sample into each channel layer. Then, we characterized the focusing performance in the MFIM chip over a wide flow-rate range. The experimental results showed that our IM-centrifuge was able to focus various-sized particles/cells to achieve volume reduction. The sample distribution strategy also effectively ensured identical focusing and concentration performances in different cores. Finally, our IM-centrifuge was successfully applied to concentrate microalgae cells with irregular shapes and highly polydisperse sizes. Thus, our IM-centrifuge holds the potential to be employed as a low-cost, high-throughput centrifuge for disposable use in low-resource settings., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The inertial microfluidic technique, as a powerful new tool for accurate cell/particle separation based on the hydrodynamic phenomenon, has drawn considerable interest in recent years. Despite numerous microfluidic techniques of particle separation, there are few articles in the literature on separation techniques addressing external outlet geometry to increase the throughput efficiency and purity. In this work, we report on a spiral inertial microfluidic device with high efficiency (>98%). Herein, we demonstrate how changing the outlet geometry can improve the particle separation throughput. We present a complete separation of 4 and 6 μm from 10 μm particles potentially applicable to separate microalgae (Tetraselmis suecica from Phaeodactylum tricornutum). Two spiral microchannels with the same cross section dimension but different outlet geometry were considered and tested to investigate the particle focusing behavior and separation efficiency. As compared with particle focusing observed in channels with a simple outlet, the particle focusing in a modified outlet geometry appears in a more successful focusing manner with complete separation. This simple approach of particle separation makes it attractive for lab-on-a-chip devices for continuous extraction and filtration of a wide range of cell/particle sizes., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Introduction: Marine planktonic communities are complex microbial consortia often dominated by microscopic algae. The taxonomic identification of individual phytoplankton cells usually relies on their morphology and demands expert knowledge. Recently, a live single-cell mass spectrometry (LSC-MS) pipeline was developed to generate metabolic profiles of microalgae., Objective: Taxonomic identification of diverse microalgal single cells from collection strains and plankton samples based on the metabolic fingerprints analyzed with matrix-free laser desorption/ionization high-resolution mass spectrometry., Methods: Matrix-free atmospheric pressure laser-desorption ionization mass spectrometry was performed to acquire single-cell mass spectra from collection strains and prior identified environmental isolates. The computational identification of microalgal species was performed by spectral pattern matching (SPM). Three similarity scores and a bootstrap-derived confidence score were evaluated in terms of their classification performance. The effects of high and low-mass resolutions on the classification success were evaluated., Results: Several hundred single-cell mass spectra from nine genera and nine species of marine microalgae were obtained. SPM enabled the identification of single cells at the genus and species level with high accuracies. The receiver operating characteristic (ROC) curves indicated a good performance of the similarity measures but were outperformed by the bootstrap-derived confidence scores., Conclusion: This is the first study to solve taxonomic identification of microalgae based on the metabolic fingerprints of the individual cell using an SPM approach.

Separating specific cell phenotypes from a heterotypic mixture is a critical step in many research projects. Traditional methods usually require a large sample volume and a complex preparation process that may alter cell property during the sorting process. Here we present the use of electrical impedance as an indicator of cell health and for identifying specific microalgal phenotypes. We developed a microfluidic platform for measuring electrical impedance at different frequencies using the bacterium-sized green alga Picochlorum SE3. The cells were cultured under different salinity conditions and sampled at four different time points. Our results demonstrate the utility of electrical impedance as an indicator of cell phenotype by providing results that are consistent with known changes in cell size and physiology. Outliers in the cell data distribution are particularly useful because they represent phenotypes that have the ability to maintain size and/or membrane ionic permeability under prolonged salt stress. This suggests that our device can be used to identify and sort desired (e.g., experimentally evolved, mutant) cell phenotypes based on their electrical impedance properties.

The development of nanotechnology and the upsurge of interest in titanium dioxide (TiO 2 ) nanoparticles, especially the anatase and rutile crystalline phases, in consumer products such as paint and sunscreen, has polluted the aquatic environment and had adverse effects on living organisms, especially algae. Microalgae help to preserve the aquatic ecosystem. Accordingly, the present study investigated the effects of anatase and rutile TiO 2 nanoparticles on the growth, photosynthetic pigment (chlorophyll), photosynthesis, and respiration rate of two algae species, Dunaliella salina (at NaCl concentrations of 1.5 and 0.5 M) and Dunaliella tertiolecta (at NaCl concentrations of 0.5 and 0.17 M). Treatment with 50, 100, 150, and 200 ppm of TiO 2 and nano-TiO 2 revealed that nano-TiO 2 inhibited the growth and decreased the specific growth rate, chlorophyll, and photosynthesis of both algal species. The rate of decrease was significantly lower at higher concentrations of NaCl in both species; however, the greatest significant difference was observed at lower concentrations of NaCl in the anatase phase. The respiration rate increased for 2 weeks but, especially at lower concentrations of NaCl, the rate of increase declined at higher concentrations after exposure to both substances, especially in the anatase phase. The findings reveal that nano-TiO 2 has a toxic effect on Dunaliella algae and its effect depends on the concentration of NaCl. The toxic effect was shown to decrease at higher concentrations of NaCl.

Laser capture microdissection is a valuable technique in individually isolating single cells whether in tissue networks or deposited from a cell suspension. New developments have enabled coupling of laser capture microdissection with mass spectrometry via liquid vortex capture sampling probe. This enables online metabolic profiling of sectioned cells. Here, we describe the protocol used to deposit, isolate, and individually chemically characterize single Allium cepa and Chlamydomonas reinhardtii cells by laser capture microdissection-liquid vortex capture mass spectrometry.

In situ monitoring in microbial bioprocesses is mostly restricted to chemical and physical properties of the medium (e.g.,pH value and the dissolved oxygen concentration). Nevertheless, the morphology of cells can be a suitable indicator for optimal conditions, since it changes with dependence on the growth state, product accumulation and cell stress. Furthermore, the single-cell size distribution provides not only information about the cultivation conditions, but also about the population heterogeneity. To gain such information, a photo-optical in situ microscopy device 1 was developed to enable the monitoring of the single-cell size distribution directly in the cell suspension in bioreactors. An automated image analysis is coupled to the microscopy based on a neural network model, which is trained with user-annotated images. Several parameters, which are gained from the captures of the microscope, are correlated to process relevant features of the cells, like their metabolic activity. Until now, the presented in situ microscopy probe series was applied to measure the pellet size in filamentous fungi suspensions. It was used to distinguish the single-cell size in microalgae cultivation and relate it to lipid accumulation. The shape of cellular particles was related to budding in yeast cultures. The microscopy analysis can be generally split into three steps: (i) image acquisition, (ii) particle identification, and (iii) data analysis, respectively. All steps have to be adapted to the organism, and therefore specific annotated information is required in order to achieve reliable results. The ability to monitor changes in cell morphology directly in line or on line (in a by-pass) enables real-time values for monitoring and control, in process development as well as in production scale. If the off line data correlates with the real-time data, the current tedious off line measurements with unknown influences on the cell size become needless.

From a high-potential biomass perspective, microalgae have recently attracted considerable attention due to their extensive application in many areas. Although studies searching for algal species with extensive application potential are ongoing, technical development for their assessment and maintenance of quality in culture are also critical and inescapable challenges. Considering the sensitivity of microalgae to environmental changes, management of algal quality is one of the top priorities for industrial applications. Helping substitute for conventional methods such as manual hemocytometry, turbidity, and spectrophotometry, this review presents an image-based, automated cell counter with a fluorescence filter to measure chlorophyll autofluorescence emitted by algae. Capturing chlorophyll-bearing cells selectively, the device accomplished precise qualification of algal numbers. The results for cell density using the device with fluorescence detection were almost identical to those obtained using hemocytometry. The automated functions of the device allow operators to reduce working hours, for not only cell density analysis but simultaneous multiparametric analysis such as cell size and algal status based on chlorophyll integrity. The automated device boldly supports further development of algal application and might contribute to opening up more avenues in the microalgal industry.

There are a huge number, and abundant types, of microalgae in the ocean; and most of them have various values in many fields, such as food, medicine, energy, feed, etc. Therefore, how to identify and separation of microalgae cells quickly and effectively is a prerequisite for the microalgae research and utilization. Herein, we propose a microfluidic system that comprised microalgae cell separation, treatment and viability characterization. Specifically, the microfluidic separation function is based on the principle of deterministic lateral displacement (DLD), which can separate various microalgae species rapidly by their different sizes. Moreover, a concentration gradient generator is designed in this system to automatically produce gradient concentrations of chemical reagents to optimize the chemical treatment of samples. Finally, a single photon counter was used to evaluate the viability of treated microalgae based on laser-induced fluorescence from the intracellular chlorophyll of microalgae. To the best of our knowledge, this is the first laboratory prototype system combining DLD separation, concentration gradient generator and chlorophyll fluorescence detection technology for fast analysis and treatment of microalgae using marine samples. This study may inspire other novel applications of micro-analytical devices for utilization of microalgae resources, marine ecological environment protection and ship ballast water management.

Collecting microalgae from water with less energy and cost is significant to gain economic profit from microalgae harvesting and processing. Foam separation has certain advantages including low energy consumption, simple operation and easy maintenance of the equipment. Natural surfactants, compared to traditional surfactants, were used to harvest and separate the freshwater microalgae Desmodesmus brasiliensis by foam separation. Results showed a recovery percentage of 93.6% and an enrichment ratio of 23.1 with the natural surfactant cocamidopropyl betaine (CAPB), suggesting that this low-cost surfactant can be applied to microalgae biomass recovery on a commercial scale using foam separation with no negative effect on the content of microalgae chlorophyll, carotenoid or protein.

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.

There is increasing concern about how microplastics (MPs) are impacting marine ecosystems. In particular, studies on how MPs impact microalgae are required because of the abundance of MPs and importance of green microalgae as primary producers. This study investigated how MPs that are larger (200 μm) than algal cells impact them, using the marine microalga Dunaliella salina as the test species. The microalga was exposed to polyethylene MPs for 6 days. Of interest, the growth and photosynthetic activity of D. salina was enhanced with exposure to MPs, while cell morphology (size and granularity) was not impacted. This phenomenon might be explained by trace concentrations of additive chemicals (endocrine disruptors, phthalates, stabilizers) that possibly leached from MPs promoting the growth and photosynthetic activity of D. salina. We also confirmed that MP size contributes towards determining how plastics affect microalgae. Specifically, as MP size shrinks compared to algal cell size, MPs have increasingly adverse effects. MPs of very small size (like nanoplastics) induce particularly adverse effects on algae. Further studies are required to establish the relationship between algal cell size and MP size., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Microplastics (MPs) could pose potential risks to microalgae, the primary producer of marine ecosystems. Currently, few studies focus on the interaction of aged MPs with other pollutants and their toxic effects to microalgae. Therefore, the present study aimed to investigate i) the aging of microplastics polyvinyl chloride (mPVC) in simulated seawater and the changes in physical and chemical properties; ii) the effects of single mPVC (virgin and aged) and copper on microalgae Chlorella vulgaris; and iii) the interaction of aged mPVC and copper and the oxidative stress towards C. vulgaris. In this study, some wrinkles, rough and fractured surface textures can be observed on the aged mPVC, accompanying with increased hydroxyl groups and aromatic carbon-carbon double bond but decreased carbon hydrogen bond. It was found that single virgin or aged mPVC at low concentration (10 mg/L) had significant inhibition on the growth of C. vulgaris but no inhibition at higher concentration (100, 1,000 mg/L), which can be reasonably explained by the aggregation and precipitation of mPVC at high concentration. The aging of mPVC inhibited the growth of C. vulgaris with the maximum growth inhibition ratio (IR) of 35.26% as compared with that of virgin mPVC (IR = 28.5%). However, the single copper could significantly inhibit the growth of C. vulgaris and the inhibitory effects increased with concentration (0.2, 0.5, 1.0 mg/L). Furthermore, both the single aged mPVC (10 mg/L) and copper (0.5 mg/L) caused serious cell damage, although the concentration of superoxide dismutase (SOD) and the intracellular malonaldehyde (MDA) increased. In contrast to single treatment, the growth of C. vulgaris can be enhanced by the combined group with copper (0.5 mg/L) and aged mPVC (10 mg/L)., (Copyright © 2019. Published by Elsevier B.V.)

Microalgae cells have been recognized as a promising sustainable resource to meet worldwide growing demands for renewable energy, food, livestock feed, water, cosmetics, pharmaceuticals, and materials. In order to ensure high-efficiency and high-quality production of biomass, biofuel, or bio-based products, purification procedures prior to the storage and cultivation of the microalgae from contaminated bacteria are of great importance. The present work proposed and developed a simple, sheathless, and efficient method to separate microalgae Chlorella from bacteria Bacillus Subtilis in a straight channel using the viscoelasticity of the medium. Microalgae and bacteria migrate to different lateral positions closer to the channel centre and channel walls respectively. Fluorescent microparticles with 1 μm and 5 μm diameters were first used to mimic the behaviours of bacteria and microalgae to optimize the separating conditions. Subsequently, size-based separation in Newtonian fluid and in viscoelastic fluid in straight channels with different aspect ratios was compared and demonstrated. Under the optimal condition, the removal ratio for 1 μm microparticles and separation efficiency for 5 μm particles can reach up to 98.28% and 93.85% respectively. For bacteria and microalgae cells separation, the removal ratio for bacteria and separation efficiency for microalgae cells is 92.69% and 100% respectively. This work demonstrated the continuous and sheathless separation of microalgae from bacteria for the first time by viscoelastic microfluidics. This technique can also be applied as an efficient and user-friendly method to separate mammalian cells or other kinds of cells.

Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood., Competing Interests: Conflict of interest statement: Y.O., Y.S., Y.W., and K.G. are inventors of a patent application covering SRS imaging with a wavelength-switched laser. All other authors declare no conflict of interest., (Copyright © 2019 the Author(s). Published by PNAS.)

Microalgal harvesting and dewatering are the main bottlenecks that need to be overcome to tap the potential of microalgae for production of valuable compounds. Water surface-floating microalgae form robust biofilms, float on the water surface along with gas bubbles entrapped under the biofilms, and have great potential to overcome these bottlenecks. However, little is known about the molecular mechanisms involved in the water surface-floating phenotype. In the present study, we analysed the genome sequence of a water surface-floating microalga Chlorococcum sp. FFG039, with a next generation sequencing technique to elucidate the underlying mechanisms. Comparative genomics study with Chlorococcum sp. FFG039 and other non-floating green microalgae revealed some of the unique gene families belonging to this floating microalga, which may be involved in biofilm formation. Furthermore, genetic transformation of this microalga was achieved with an electroporation method. The genome information and transformation techniques presented in this study will be useful to obtain molecular insights into the water surface-floating phenotype of Chlorococcum sp. FFG039.

Intelligent image-activated cell sorting (iIACS) is a machine-intelligence technology that performs real-time intelligent image-based sorting of single cells with high throughput. iIACS extends beyond the capabilities of fluorescence-activated cell sorting (FACS) from fluorescence intensity profiles of cells to multidimensional images, thereby enabling high-content sorting of cells or cell clusters with unique spatial chemical and morphological traits. Therefore, iIACS serves as an integral part of holistic single-cell analysis by enabling direct links between population-level analysis (flow cytometry), cell-level analysis (microscopy), and gene-level analysis (sequencing). Specifically, iIACS is based on a seamless integration of high-throughput cell microscopy (e.g., multicolor fluorescence imaging, bright-field imaging), cell focusing, cell sorting, and deep learning on a hybrid software-hardware data management infrastructure, enabling real-time automated operation for data acquisition, data processing, intelligent decision making, and actuation. Here, we provide a practical guide to iIACS that describes how to design, build, characterize, and use an iIACS machine. The guide includes the consideration of several important design parameters, such as throughput, sensitivity, dynamic range, image quality, sort purity, and sort yield; the development and integration of optical, microfluidic, electrical, computational, and mechanical components; and the characterization and practical usage of the integrated system. Assuming that all components are readily available, a team of several researchers experienced in optics, electronics, digital signal processing, microfluidics, mechatronics, and flow cytometry can complete this protocol in ~3 months.

Quick algae harvesting methodologies relating optimum flocculent dose (D Opt. ), percentage harvesting efficiency (%HE) and percentage water recovery (%W Recovery ) to the in-situ hydrodynamic properties of water-algae systems are presented. Flocculation of three microalgae in single and sequential steps, using chemical (polymer and ferric chloride) and biological (egg shells) flocculants, was studied. Zeta potential and pH analysis were completed to further understand the flocculation mechanism. Polymer at D Opt. of 7.0 g/kgDS resulted in W Recovery of 90% and %HE of 96.7%. Lower %HE (92.1), %W Recovery (79) and noticeable algal cells deformation was observed for ferric chloride at D Opt. of 7.0 g/kg DS. Bio-flocculant conserved algal structure and resulted in %HE of 96.2 and %W Recovery of 90 at D Opt. of 5.4 g/kgDS. Significant % HE of 99.8, %W Recovery of 99.8%, and up to 95% reduction in D Opt. were achieved in sequential flocculation. The results established the effectiveness and suitability of sequential/ bio-flocculation for algae harvesting., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

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.)

Microalgae adhesion plays a critical role in developing effective photobioreactors for large-scale production of microalgae biofuel. This study focused on elucidating the influencing mechanism of liquid medium pH on microalgae adhesion by identifying the dominant interactions between cell and substratum using a criterion. Herein, the adhesion of three microalgae onto two substrata at a series of pH was observed using a flow chamber. The results indicated that the adhesion of freshwater Chlorella sp. onto PVC and glass and marine Chlorella sp. and N. oculata onto glass decreased with increasing pH, because these adhesions were dominated by the EL interaction, and the pH would influence the adhesion primarily by affecting the ζ potential of the cell and substratum. Whereas, the adhesion of marine Chlorella sp. and N. oculata onto PVC increased with increasing pH, because these adhesions were dominated by Lewis acid-base (AB) interaction, and the pH would influence the adhesion primarily by affecting the components of surface free energy of cell. The study demonstrated that the influencing mechanism of pH on adhesion can be conclusively elucidated by identifying the dominant interaction between the cell and the surface, and may have significant implications for predicting cell adhesion in various applications., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Increasing numbers of studies in the past few decades have demonstrated vertically-oriented nanoneedles arrays (NNAs) as innovative tools to interrogate and manipulate biological cells, where the needles are inserted into the cells as functional probes for high-throughput detection and biomolecule delivery. However, majority of these studies use mammalian cells: leaving NNA application to plant cells still in its infancy and largely unexplored. This paper highlights our contributions in exploring the utility of NNAs to microalgae - a diverse group of aquatic, photosynthetic organisms studied intensively as bio-factories for producing high-value-added products such as fuels and pharmaceuticals. Microalgal strain development processes have long suffered from the hard cell wall that surrounds the cell and inhibits delivery of foreign materials into the cell. Conically-shaped, metallic NNAs were developed with template synthesis that successfully penetrate through the cell wall barrier and achieve material injection - using the widely studied model microalga, Chlamydomonas reinhardtii. Earlier works from mammalian cells are introduced and discussed to clarify the framework established in this field, while recent studies of both mammalian and microalgal cells are also referenced to examine the trends, challenges, and future perspectives of NNA application to microalgae., (© 2019 The Chemical Society of Japan & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Microalgae are an ubiquitous and powerful driver of geochemical cycles which have formed Earth's biosphere since early in the evolution. Lately, microalgal research has been strongly stimulated by economic potential expected in biofuels, wastewater treatment, and high-value products. Similar to bacteria and other microorganisms, most work so far has been performed on the level of suspensions which typically contain millions of algal cells per millilitre. The thus obtained macroscopic parameters average cells, which may be in various phases of their cell cycle or even, in the case of microbial consortia, cells of different species. This averaging may obscure essential features which may be needed for the correct understanding and interpretation of investigated processes. In contrast to these conventional macroscopic cultivation and measuring tools, microfluidic single-cell cultivation systems represent an excellent alternative to study individual cells or a small number of mutually interacting cells in a well-defined environment. A novel microfluidic photobioreactor was developed and successfully tested by the photoautotrophic cultivation of Chlorella sorokiniana. The reported microbioreactor facilitates automated long-term cultivation of algae with controlled temperature and with an illumination adjustable over a wide range of photon flux densities. Chemical composition of the medium in the microbioreactor can be stabilised or modulated rapidly to study the response of individual cells. Furthermore, the algae are cultivated in one focal plane and separate chambers, enabling single-cell level investigation of over 100 microcolonies in parallel. The developed platform can be used for systematic growth studies, medium screening, species interaction studies, and the thorough investigation of light-dependent growth kinetics., Competing Interests: The authors have declared that no competing interests exist.

The composition of polyunsaturated fatty acids (PUFAs) in triacylglycerols (TAGs) is key to health benefits and for oil applications, yet the underlying genetic mechanism remains poorly understood. In this study, by in silico, ex vivo, and in vivo profiling of type-2 diacylglycerol acyltransferases (DGAT2s) in Nannochloropsis oceanica we revealed two novel PUFA-preferring enzymes that discriminate individual PUFA species in TAG assembly, with NoDGAT2J for linoleic acid (LA) and NoDGAT2K for eicosapentaenoic acid (EPA). The LA and EPA composition of TAG molecules is mediated in vivo via the functional partitioning between NoDGAT2J and 2K, both of which are localized in the chloroplast envelope. By modulating transcript abundance of the DGAT2s, an N. oceanica strain bank was created, where proportions of LA and EPA in TAG vary by 18.7-fold (between 0.21% and 3.92% dry weight) and 34.7-fold (between 0.09% and 3.12% dry weight), respectively. These findings lay the foundation for producing designer TAG molecules with tailored health benefits or for biofuel applications in industrial microalgae and higher-plant crops., (Copyright © 2018 The Author. Published by Elsevier Inc. All rights reserved.)

In this study, an innovative helical coagulation reactor (HCR) was developed for harvesting microalgae by sedimentation with polyaluminium chloride (PAC). The effects of construction and hydrodynamic characteristics on harvesting performance were investigated. Results showed that a higher harvesting efficiency, 96.37%, was achieved for the large and compact flocs generated by the HCR, and the settling rate of flocs was substantially influenced by the velocity gradient (G) and the Reynolds number (Re). When the Reynolds number closed to the transition between laminar and turbulent flow (4000), the flocs settled faster (20.51 m h -1 ), although settling slowed as the Reynolds number increased further because of ruptured flocs. The settling rate of flocs could be further improved to 23.27 m h -1  by a pulse flow field, mainly due to larger and more compact flocs forming in the plug pipe flow. Furthermore, a comparative investigation of a mechanically agitated vessel and the HCR with the same Camp number (Gt) showed that the HCR achieved higher settling rates and a shorter residence time than those with a mechanical agitator. The HCR provided a uniform dissipation of energy and high velocity gradient while avoiding electrical and mechanical energy consumption, suggesting this reactor is an efficient and economic option for microalgae harvesting., (Copyright © 2018 The Society for Biotechnology, Japan. Published by 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.)

Nowadays, the pretreatment of wastewater prior to discharge is very important in various industries as the wastewater without any treatment contains high organic pollution loads that would pollute the receiving waterbody and potentially cause eutrophication and oxygen depletion to aquatic life. The reuse of seafood wastewater discharge in microalgae cultivation offers beneficial purposes such as reduced processing cost for wastewater treatment, replenishing ground water basin as well as financial savings for microalgae cultivation. In this paper, the cultivation of Chlorella vulgaris with an initial concentration of 0.01 ± 0.001 g⋅L -1 using seafood sewage discharge under sunlight and fluorescent illumination was investigated in laboratory-scale without adjusting mineral nutrients and pH. The ability of nutrient removal under different lighting conditions, the metabolism of C. vulgaris and new medium as well as the occurrence of auto-flocculation of microalgae biomass were evaluated for 14 days. The results showed that different illumination sources did not influence the microalgae growth, chemical oxygen demand (COD) and biochemical oxygen demand (BOD) significantly. However, the total nitrogen (total-N) and total phosphorus (total-P) contents of microalgae were sensitive to the illumination mode. The amount of COD, BOD, total-N and total-P were decreased by 88%, 81%, 95%, and 83% under sunlight mode and 81%, 74%, 79%, and 72% under fluorescent illumination, respectively. Furthermore, microalgae were auto-flocculated at the final days of cultivation with maximum biomass concentration of 0.49 ± 0.01 g⋅L -1 , and the pH value had increased to pH 9.8 ± 0.1 under sunlight illumination., (Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The composition of the ship's ballast water is complex and contains a large number of microalgae cells, bacteria, microplastics, and other microparticles. To increase the accuracy and efficiency of detection of the microalgae cells in ballast water, a new microfluidic chip for continuous separation of microalgae cells based on alternating current dielectrophoresis was proposed. In this microfluidic chip, one piece of 3-dimensional electrode is embedded on one side and eight discrete electrodes are arranged on the other side of the microchannel. An insulated triangular structure between electrodes is designed for increasing the inhomogeneity of the electric field distribution and enhancing the dielectrophoresis (DEP) force. A sheath flow is designed to focus the microparticles near the electrode, so as to increase the suffered DEP force and improve separation efficiency. To demonstrate the performance of the microfluidic separation chip, we developed two species of microalgae cells (Platymonas and Closterium) and a kind of microplastics to be used as test samples. Analyses of the related parameters and separation experiments by our designed microfluidic chip were then conducted. The results show that the presented method can separate the microalgae cells from the mixture efficiently, and this is the first time to separate two or more species of microalgae cells in a microfluidic chip by using negative and positive DEP force simultaneously, and moreover it has some advantages including simple operation, high efficiency, low cost, and small size and has great potential in on-site pretreatment of ballast water., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)