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Given its profound disservice, a bacteria-microalgae-fungi combined system was designed to treat kitchen waste. Firstly, a new type of microbial agent homemade compound microorganisms (HCM) (composed of Serratia marcescens, Bacillus subtilis and other 11 strains) with relatively high bio-security were developed for pretreating kitchen waste, and HCM efficiently degraded 85.2 % cellulose, 94.3 % starch, and 59.0 % oil. HCM also accomplished brilliantly the initial nutrients purification and liquefaction conversion of kitchen waste. Under mono-culture mode (fungi and microalgae were inoculated separately in the pre - and post-stages) and co-culture mode (fungi and microalgae were inoculated simultaneously in the early stage), microalgae-fungi consortia were then applied for further water purification and resource utilization of kitchen waste liquefied liquid (KWLL) produced in the pretreatment stage. Two kinds of microalgae-fungi consortia (Chlorella sp. HQ and Chlorella sp. MHQ2 form consortia with pellet-forming fungi Aspergillus niger HW8-1, respectively) removed 79.5-83.0 % chemical oxygen demand (COD), 44.0-56.5 % total nitrogen (TN), 90.3-96.4 % total phosphorus (TP), and 64.9-71.0 % NH4+-N of KWLL. What's more, the microalgae-fungi consortia constructed in this study accumulated abundant high-value substances at the same time of efficiently purifying KWLL. Finally, in the biomass harvesting stage, pellet-forming fungi efficiently harvested 81.9-82.1 % of microalgal biomass in a low-cost manner through exopolysaccharides adhesion, surface proteins interaction and charge neutralization. Compared with conventional microalgae-bacteria symbiosis system, the constructed bacteria-microalgae-fungi new-type combined system achieves the triple purpose of efficient purification, resource utilization, and biomass recovery on raw kitchen waste through the trilogy strategy, providing momentous technical references and more treatment systems selection for future kitchen waste treatment.

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The efficient and economical treatment of wastewater using microalgae has attracted much attention. However, harvesting microalgae cells from treated wastewater remains challenging. In the present study, a Chlorella vulgaris suspension containing filamentous fungi Aspergillus niger and Chaetomium gracile was successfully used to construct a self-flocculating system, with a microalgae flocculation efficiency of 99.6% achieved by gravity sedimentation within 4 h. The diameter of fungi played an important role in determining flocculation efficiency, and the optimal particle size was 10 mm. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) results indicated that the sweeping action of fungal mycelia and the interaction between the functional groups of fungi and the C. vulgaris surface contributed to improve flocculation. Co-cultivation of C. vulgaris and fungi could effectively remove 83.53%, 94.45% and 76.88% of total phosphorus, total nitrogen and chemical oxygen demand, respectively, from the sludge leachate from a sugar mill. The fungal-algal biomass reached 5.75 g/L. Herein, the constructed self-flocculation system had coupled efficient flocculation of C. vulgaris with removal of pollutants from wastewater in a short period of time, and providing a green, pollution-free, low-cost method for simultaneous wastewater treatment and the production of high quality biomass.

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The use of chitosan as a flocculant has become a topic of interest over the years due to its positively charged polymer and biodegradable and non-toxic properties. However, most studies only focus on microalgae and wastewater treatment. This study provides crucial insight into the potential of using chitosan as an organic flocculant to harvest lipids and docosahexaenoic acid (DHA-rich Aurantiochytrium sp. SW1 cells by examining the correlation of flocculation parameters (chitosan concentration, molecular weight, medium pH, culture age, and cell density) toward the flocculation efficiency and zeta potential of the cells. A strong correlation between the pH and harvesting efficiency was observed as the pH increased from 3, with the optimal flocculation efficiency of >95% achieved at a chitosan concentration of 0.5 g/L at pH 6 where the zeta potential was almost zero (3.26 mV). The culture age and chitosan molecular weight have no effect on the flocculation efficiency but increasing the cell density decreases the flocculation efficiency. This is the first study to reveal the potential of chitosan to be used as a harvesting alternative for thraustochytrid cells.

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Biofilm-based algal technologies have gained popularity due to higher biomass productivity, efficient harvesting, and water-saving over suspended growth systems. A rotating attached system was designed to assess the biofilm-forming capacity of different isolated microalgal strains from the Persian Gulf. Four microalgal strains, including two Chlorella sp., one Picochlorum sp. and one filamentous cyanobacterium Desmonostoc sp. were cultivated on four carriers: jute, cotton, yarn and nylon. The carriers' physicochemical surface characteristics and attachment effects, like contact angle, were investigated. The incorporated biomass and exopolysaccharides (EPS) content in the suspended and biofilm system was calculated and compared. The results showed that the cyanobacterium strain had the biofilm formation capability on both jute and cotton in the attached cultivation system. Under the same culture conditions, the biomass productivity on jute and cotton carriers was significantly higher (4.76 and 3.61 g m- 2 respectively) than the growth in aqueous suspension (1.19 g m- 2 d- 1). The greatest incorporated exopolysaccharides amount was observed on jute (43.62 ± 4.47%) and the lowest amount was obtained from the growth on positive charge yarn (18.62 ± 1.88%). This study showed that in comparison with planktonic growth, the colonization of cyanobacterial cells and subsequent production of extracellular matrix and biofilm formation can lead to increased biomass production.

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Microalgal biomass harvesting and cell disruption are the main bottlenecks for downstream processing of microalgae such as high-value bioproducts extraction and biofuels production. In this study, we evaluated the performance of dual flocculation between cationic surfactants and bio-polymer of chitosan for simultaneous biomass harvesting and bioproducts extraction from Chlorella sorokiniana microalgae. First, the effects of individual natural flocculants of chitosan and two cationic surfactants: cetyltrimethylammonium bromide (CTAB) and dodecyltrimethylammonium bromide (DTAB) on biomass harvesting were studied. Next, the synergistic effect of dual flocculation between the cationic surfactants and chitosan on harvesting efficiency, time and flocculant dosage was investigated. Finally, we evaluated the potential of high value bioproducts extraction from microalgae after the individual and dual flocculation processes. Zeta potential analysis and microscopic images were employed to achieve mechanistic understanding. Maximum biomass harvesting efficiencies of 85 %, 88 % and 78 % were achieved using individual flocculants of chitosan, CTAB and DTAB, under their optimum dosages of 100, 400 and 4000 mg/L, respectively. A significant synergistic effect of dual flocculation between chitosan (C) and cationic surfactants on biomass harvesting efficiency (CTAB-C: 99 % and DTAB-C: 97 %), settling time (CTAB-C: 2 min and DTAB-C: 5 min) and optimum dosage of surfactants (CTAB-C: 100 mg/L and DTAB-C: 1000 mg/L) was observed. The synergistic effect was associated with multiple flocculation mechanisms of charge neutralization and bridging induced by cationic surfactants and chitosan, respectively. Furthermore, bioproducts recovery efficiencies of 12 %, 25 % and 15 % of cell dry weight were achieved for protein, carbohydrate and lipid, respectively by using dual flocculation of CTAB surfactant and chitosan at much lower dosage of 100 mg/L.

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A novel method of one-step co-cultivation and harvesting of microalgae and fungi, for efficient starch wastewater treatment and high-value biomass production was developed. By combination of Aspergillus oryzae and Chlorella pyrenoidosa, nutrients in wastewater could be converted to useful microbial biomass, while the wastewater was purified. Moreover, the microalgae C. pyrenoidosa could gradually be encapsulated in fungal pellets which promoted the biomass harvesting. The free algal cells could be completely harvested by fungal pellets within 72 h. The synergistic effects between them greatly improved the removal efficiencies of main pollutants as the removal efficiency of COD, TN, and TP reached 92.08, 83.56, and 96.58 %, respectively. In addition, the final biomass concentration was higher than that of individual cultures. The protein and lipid concentration was also significantly improved and reached 1.92 and 0.99 g/L, respectively. This study provides a simple and efficient strategy for simultaneous wastewater treatment and high-value biomass production.

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Immobilized microalgae are good for wastewater treatment and biomass production. This study investigated treatment efficiency of a continuously operated system employing immobilized microalgae for secondary effluent of wastewater treatment plants, as well as the effectiveness on induction of valuable products, harvesting and dewatering of microalgae biomass. Under semi-continuous operation condition, microalgal dry weight increased 4.75 times within 2 d, associated with the highest removal rate of ammonia and phosphate at 28.95 mg/L·d and 4.83 mg/L·d, respectively. An immobilized microalgae membrane bioreactor (iMBR) was continuously operated for a month. The harvested immobilized microalgae beads were transferred to induction stage to obtain 4.5 times increase of lipid content per cell within 2 d. Immobilized microalgae performed 1.9 cm/s settling velocity and 97% water removal efficiency around 40 °C. A prospective integrated process on resource recovery and carbon neutrality was proposed for wastewater treatment, induction, harvesting and dewatering of immobilized microalgae cells.

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The most frequently used method to harvest microalgae on an industrial scale is centrifugation, although this has very high energy costs. To reduce these costs, a continuous electrocoagulation process for harvesting Chlorella vulgaris was developed and tested using a pilot-scale 111 L working volume device consisting of an electrolyser with iron electrodes, aggregation channel and lamellar settler. The flow rate of the microalgal suspension through the device was 240 L/h. When using controlled cultivation and subsequent electrocoagulation, a high harvesting efficiency (above 85%), a low Fe contamination in the harvested biomass (<4 mg Fe/g dry biomass, a harvested biomass complied with legislative requirements for food) and significant energy savings were achieved. When comparing electrocoagulation and subsequent centrifugation with the use of centrifugation alone, energy savings were 80 % for a biomass harvesting concentration of 0.23 g/L. Electrocoagulation was thus proven to be a feasible pre-concentration method for harvesting microalgae.

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Biodiesel produced from microalgae is a potential alternative due to the high growth rate of microalgae, the possibility of using nonarable land, and high lipid accumulation rate. Microalgae cultivation, cell harvesting and disruption are the important steps before lipid extraction for the biodiesel. In the co-submission article, the details of the whole process cannot be clearly explained. In this regard, we present the details of methods on parameter of photo-bioreactor for cultivating microalgae, flocculation tests to determine optimal flocculant dosage in harvesting, parameter of Dimethyl ether (DME) subcritical extraction device and full-factorial design for investigating the influence of extraction time, initial water content and DME dosage on the extraction performance. It will allow researchers to reproduce these experiments. • The method shows a cell disruption assisted lipid extraction by subcritical dimethyl ether. • Model is built from full-factorial design to investigate multi-factor influence. • Differential scanning calorimetry can be applicable to measure free water content.

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Cyanobacteria and microalgae are considered as interesting feedstocks for either the production of high value bio-based compounds and biofuels or wastewater treatment. Nevertheless, the high costs of production, mainly due to the harvesting process, hamper a wide commercialization of industrial cyanobacteria and microalgae based products. Recent studies have found in autoflocculation and bioflocculation promising spontaneous processes for a low-cost and environmentally sustainable cyanobacteria and microalgae biomass harvesting process. In the present work, bioflocculation process has been studied for three different inocula: filamentous cyanobacteria, microalgae and their mixture. Their cultivation has been conducted in batch mode using two different cultivation media: synthetic aqueous solution and urban wastewater. The removal of nutrients and flocculation process performance were monitored during the entire cultivation time. Results have proved that bioflocculation and sedimentation processes occur efficiently for filamentous cyanobacteria cultivated in synthetic aqueous solution, whereas such processes are less efficient in urban wastewater due to the specific characteristics of this medium that prevent bioflocculation to occur. Besides different efficiencies associated to cultivation media, this work highlighted that bioflocculation of sole microalgae is not as effective as when they are cultivated together with filamentous cyanobacteria.

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Flocculation is a low-cost harvesting technique for microalgae biomass production, but flocculation efficiency is species dependent. In this study, we investigated the efficacy of two synthetic (polyacrylamide) and one natural (chitosan) flocculants against three algal species: the cyanobacterium Synechocystis sp., the freshwater Chlorella vulgaris, and the marine Phaeodactylum tricornutum at laboratory and pilot scales to evaluate harvesting efficiency, biomass integrity and media recycling. Growth phase affected the harvesting efficiency of the eukaryotic microalgae. The flocculation was optimal at stationary phase with high flocculation efficiency achieved using polyacrylamides at 24-36 mg/g dry weight. The effect of the flocculants on the harvested biomass was investigated. The flocculated Synechocystis sp. showed a higher proportion of compromised cells compared to C. vulgaris and P. tricornutum likely due to differences in cell walls composition. Compromised cells could lead to the release of valuable products into the surrounding growth media during flocculation. The residual culture media was recycled once with no impact on cell growth for all treatments and algal species. The flocculation technique was demonstrated at pilot-scale using 350 L microalgal suspension, showing an efficiency of 82-90% at a polyacrylamide dosage of 6.5-10 mg/L. This efficiency and the biomass quality are comparable to the laboratory-scale results. Overall, results indicate that polyacrylamide flocculants work on a wide range of species without the need for pre-treatment. The information generated in this study can contribute to making the microalgae industry more competitive.

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The microalga Scenedesmus obtusiusculus AT-UAM efficiently captured CO2 from two flue gas streams in a hybrid photobioreactor located in a greenhouse. Uptake rates of CO2, NO, and SO2 from a formulated gas stream were 160.7 mg L-1 day-1, 0.73 mg L-1 day-1, and 1.56 mg L-1 day-1, respectively, with removal efficiencies of 100% for all gases. Exhaust gases of a motor generator were also removed with uptake rates of 111.4 mg L-1 day-1, 0.42 mg L-1 day-1, and 0.98 mg L-1 day-1, obtaining removal efficiencies of 77%, 71%, and 53% for CO2, NOx, and SO2, respectively. On average, 61% of the CO2 from both flue gas streams was assimilated as microalgal biomass. The maximum CO2 uptake rate of 182 mg L-1 day-1 was achieved for formulated flue gas flow rate above 100 mL min-1. The biomass recovery of 88% was achieved using a 20-L electro-coagulation-flotation chamber coupled to a settler with a low specific power consumption of 0.27 kWh kg-1. The photobioreactor was operated for almost 7 months without contamination of invasive species or a decrease in the activity. It is a very encouraging result for long-term operation in flue gas treatment.

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Constructed wetlands built for water treatment often need biomass harvesting to remove nutrients from the system. Usually harvesting is done during the peak growing season to maximize the amount of nutrients removed from the system. This, however, can create huge methane fluxes that escape from plant tissues to the atmosphere. We used manual chambers and eddy covariance measurements to analyze the increase in methane emissions due to the harvesting of two common wetland species, Typha spp. and Schoenoplectus spp., in two climatically different constructed wetlands in Estonia and California. In addition, we determined the biomass nutrient and carbon concentrations from harvested biomass. We found that harvesting during the summer season, e.g. June and August, resulted in a significant release of methane at both sites. At the California site, baseline median methane emissions were 217.6 nmol m-2 s-1, and harvesting resulted in increases to 395.4 nmol m-2 s-1 that decreased to baseline emission within three days. Footprint modeling demonstrated that the emission increases measured by eddy covariance were dominated by contributions from the cut area to the total footprint signal. At the Estonian site, harvesting resulted in methane increases of 15.9 nmol m-2 s-1 to 110.4 nmol m-2 s-1 in August. However, in September and October the emission was significantly lower. Plant biomass analyses showed clear temporal dynamics in terms of nutrient concentration, being highest in summer and lowest in winter. Our experiments indicate that the optimal time for aboveground biomass harvesting is at the end of the growing season before nutrient translocation to belowground plant structures begins coinciding with lowest methane emissions. Therefore, strategic planning of the harvest timing may help reduce greenhouse gas emissions from managed wetlands and thus improve their multi-faceted ecological benefit.

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Bacterial and algal floc formation was induced by inoculating three species of wastewater-derived bacteria (Melaminivora jejuensis, Comamonas flocculans, and Escherichia coli) into algal cultures (Chlorella sorokiniana). Bacterial and algal flocs formed in algal cultures inoculated with M. jejuensis and C. flocculans, and these flocs showed higher sedimentation rates than pure algal culture. The floc formed by M. jejuensis (4988.46 ± 2589.81 µm) was 10-fold larger than the floc formed by C. flocculans (488.60 ± 226.22 µm), with a three-fold higher sedimentation rate (M. jejuensis, 91.08 ± 2.32% and C. flocculans, 32.55 ± 6.33%). Biomass and lipid productivity were improved with M. jejuensis inoculation [biomass, 102.25 ± 0.35 mg/(L·day) and 57.80 ± 0.20 mg/(L·day)] compared with the productivity obtained under pure algal culture conditions [biomass, 78.00 ± 3.89 mg/(L·day) and lipids, 42.26 ± 2.11 mg/(L·day)]. Furthermore, the fatty acid composition of the biomass produced under pure algal culture conditions was mainly composed of C16:0 (43.67%) and C18:2 (45.99%), whereas the fatty acid composition of the biomass produced by M. jejuensis was mainly C16:0 (31.80%), C16:1 (24.45%), C18:1 (20.23%), and C18:2 (16.11%). These results suggest the possibility of developing an efficient method for harvesting microalgae using M. jejuensis and provide information on how to improve biomass productivity using floc-forming bacteria.

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BACKGROUND: Microalgal biomass harvesting using traditional chemicals is costly for the production of biofuels, hindering the scale-up process of the technology. Thus, the search for a cost-effective microalgal harvesting method is extremely important. Using chitosan as a natural flocculant to harvest microalgal biomass seems to be an efficient and convenient solution. Although microalgal biomass flocculation by chitosan has been reported in some previous studies, literature on the harvesting of microalgae C. vulgaris biomass using such polymer is scanty. In addition, there is limited information available on whether the usage of chitosan during the harvesting will affect downstream lipid extraction. Still, whether microalgae can be re-grown with the spent medium after chitosan flocculation is still unknown. RESULTS: In this study, microalgal biomass harvesting using chitosan as a natural flocculant and aluminum sulfate as a traditional flocculant was compared and evaluated. Optimal doses and effects on biomass sedimentation, spent medium recycling and lipid extraction were investigated. The results showed that the optimal doses for chitosan and aluminum sulfate to achieve more than 90% biomass recovery were 0.25 and 2.5 g/L, respectively. The sedimentation time of 10 min was found to be the most appropriate to remove over 90% biomass from culture. The spent medium after chitosan flocculation could be potentially recycled for the re-cultivation of microalgae, which demonstrated robust growth in comparison with those grown in the recycled medium from aluminum sulfate flocculation. The lipid content of microalgae harvested by chitosan reached 32.9, 4.6% higher than that of those harvested by aluminum sulfate, indicating that the application of the natural flocculant would not impact the downstream extraction of microalgal lipids. CONCLUSION: The results herein presented, demonstrated that chitosan is applicable for microalgal harvesting during the upscaling process. Flocculation method developed by using chitosan as a natural flocculant is a worthy microalgal harvesting method for microalgae-based biofuel production. There is hope that chitosan can be reasonably and technically realistically applied in a full-scale process for the harvesting of microalgal biomass.

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We report on the use of the polyethylenimine-based (PEI) sorbents for immobilization and harvesting of microalgae (MA) cells. Specific materials assessed were porous solid polymers from highly-branched PEI synthesized by cross-linking with epichlorohydrin (ECH) or diethylene glycol diglycidyl ether (DGDE). We estimated the effect of PEI/cross-linker ratio on the MA attachment and biocompatibility of the sorbents with the MA cells. A decrease in the cross-linker percentage resulted in the enhancement of the immobilization efficiency but impaired the cell viability as was manifested by inhibition of the photosynthetic activity of the MA cells. The rate of Chlorella vulgaris cell attachment to the sorbents with ECH was faster as compared to that of the PEI-DGDE-based polymers. The cells immobilized on the PEI-ECH sorbents showed a more profound decline in their viability (assessed via photosynthetic activity). The sorbents with 60% of DGDE were characterized by high immobilization efficiency. These sorbents supported a prolonged cultivation of the immobilized MA without impairing their viability and metabolic activity. We conclude that the sorbents with a lower percentage of DGDE (<30%) and sorbents with ECH are suitable for harvesting of the MA cells intended for immediate downstream processing, potentially without the cell desorption. To the best of our knowledge, this is the first report on successful application of PEI-based sorbents in microalgal biotechnology.

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With increasing global population and depleting resources, there is an apparent demand for radical unprecedented innovation to satisfy the basal needs of lives. Hence, non-conventional renewable energy resources like biodiesel have been worked out in past few decades. Biofuel (e.g. Biodiesel) serves to be the most sustainable answer to solve "food vs. fuel crisis". In biorefinery process, lipid extraction from oleaginous microbial lipids is an integral part as it facilitates the release of fatty acids. Direct lipid extraction from wet cell-biomass is favorable in comparison to dry-cell biomass because it eliminates the application of expensive dehydration. However, this process is not commercialized yet, instead, it requires intensive research and development in order to establish robust approaches for lipid extraction that can be practically applied on an industrial scale. This review aims for the critical presentation on cell disruption, lipid recovery and purification to support extraction from wet cell-biomass for an efficient transesterification.

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Increased market viability of harvest residues as forest bioenergy feedstock may escalate removal of coarse woody debris in managed forests. Meanwhile, many forest invertebrates use coarse woody debris for cover, food, and reproduction. Few studies have explicitly addressed effects of operational-scale woody biomass harvesting on invertebrates following clearcutting. Therefore, we measured invertebrate community response to large-scale harvest residue removal and micro-site manipulations of harvest residue availability in recently clearcut, intensively managed loblolly pine (Pinus taeda) forests in North Carolina (NC; n = 4) and Georgia (GA; n = 4), USA. We captured 39,794 surface-active invertebrates representing 171 taxonomic groups using pitfall traps situated among micro-site locations (i.e., purposefully retained piles of hardwood stems and piles of conifer stems and areas without coarse woody debris in NC; windrows and no windrows in GA). Micro-site locations were located within six, large-scale treatments (7.16-14.3 ha) in clearcuts. Large-scale treatments represented intensive harvest residue removal, 15% and 30% harvest residue retention, and no harvest residue removal. In NC, ground beetles (Coleoptera: Carabidae) and crickets (Orthoptera: Gryllidae) were three times more abundant in treatments with no harvest residue removal than those with the most intensive harvest residue removal and were reduced in treatments that retained 15% or 30% of harvest residues, although not significantly. Invertebrate taxa richness was greater at micro-site locations with retained hardwood and pine (Pinus spp.) harvest residues than those with minimal amounts of coarse woody debris. In both states, relative abundances of several invertebrate taxa, including cave crickets (Orthoptera: Rhaphidophoridae), fungus gnats (Diptera: Mycetophilidae and Sciaridae), millipedes (Diplopoda), and wood roaches (Blattodea: Ectobiidae), were greater at micro-site locations with retained harvest residues than those with minimal coarse woody debris. Intensified woody biomass harvesting without retention of ≥15% of harvest residue volume may reduce invertebrate taxa richness and abundances of some key invertebrate taxa in regenerating stands. Further, harvest residue management during and after woody biomass harvesting may be an important consideration for maintaining invertebrate diversity and conserving invertebrates that are influential in the maintenance of ecosystem function and integrity in young forests.

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Research on wastewater treatment by means of microalgal-bacterial processes has become a hot topic worldwide during the last two decades. Owing to the lower energy demand for oxygenation, the enhanced nutrient removal and the potential for resource recovery, microalgal-based technologies are nowadays considered as a good alternative to conventional activated sludge treatments in many instances. Nevertheless, biomass harvesting still constitutes one of the major challenges of microalgal-bacterial systems for wastewater treatment, which is hindered by the poor settleability of microalgal biomass. In this review, the use of microalgal-bacterial aggregates (MABAs) to overcome harvesting issues and to enhance resource recovery is presented. The fundamentals of MABAs-based technologies, the operational strategies and factors affecting the formation of MABAs, the microbiology and the methanogenic potential of the aggregates are addressed and critically discussed. The most recent findings and the challenges facing this technology towards its consolidation are also presented.

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Microalga is the only feedstock that has the theoretical potential to completely replace the energy requirements derived from fossil fuels. However, commercialization of this potential source for fuel applications is hampered due to many technical challenges with harvesting of biomass being the most energy intensive process among them. The fresh water microalgal species, Scenedesmus abundans, has been widely recognized as a potential feedstock for production of biodiesel (Mandotra et al., 2014). The present work deals with sedimentation of algal biomass using extracted chitosan and natural bentonite clay powder as flocculant. The effect of flocculant combination and different factors such as temperature, pH, and concentration of algal biomass on sedimentation rates has been analyzed. A high flocculation efficiency of 76.22 ± 7.81% was obtained at an algal biomass concentration of 1 ± 0.05 g/L for a settling time of 1 h at 50 ± 5°C with a settling velocity of 103.2 ± 0.6 cm/h and a maximum surface conductivity of 2,260 ± 2 µS/cm using an optimal design in response surface methodology (RSM). Biopolymer flocculant such as chitosan exhibited better adsorption property along with bentonite clay powder that reduced the settling time significantly.