A review on pretreatment methods for lipid extraction from microalgae biomass.

Microalgal lipids are promising and sustainable sources for the production of third-generation biofuels, foods, and medicines. A high lipid yield during the extraction process in microalgae could be influenced by the suitable pretreatment and lipid extraction methods. The extraction method itself could be attributed to the economic and environmental impacts on the industry. This review summarizes the pretreatment methods including mechanical and non-mechanical techniques for cell lysis strategy before lipid extraction in microalgae biomass. The multiple strategies to achieve high lipid yields via cell disruption techniques are discussed. These strategies include mechanical (shear forces, pulse electric forces, waves, and temperature shock) and non-mechanical (chemicals, osmotic pressure, and biological) methods. At present, two techniques of the pretreatment method can be combined to increase lipid extraction from microalgae. Therefore, the extraction strategy for a large-scale application could be further strengthened to optimize lipid recovery by microalgae.

Harvesting Aurantiochytrium sp. SW1 via Flocculation Using Chitosan: Effects of Flocculation Parameters on Flocculation Efficiency and Zeta Potential.

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.

Construction of microbial electrodialysis cells equipped with internal proton migration pathways: Enhancement of wastewater treatment, desalination, and hydrogen production.

Microbial electrodialysis cells (MEDCs) offer simultaneous wastewater treatment, water desalination, and hydrogen production. In a conventional design of MEDCs, the overall performance is retarded by the accumulation of protons on the anode due to the integration of an anion exchange membrane (AEM). The accumulation of protons reduces the anolyte pH to become acidic, affecting the microbial viability and thus limiting the charge carrier needed for the cathodic reaction. This study has modified the conventional MEDC with an internal proton migration pathway, known as the internal proton migration pathway-MEDC (IP-MEDC). Simulation tests under abiotic conditions demonstrated that the pH changes in the anolyte and catholyte of IP-MEDC were smaller than the pH changes in the anolyte and catholyte without the proton pathways. Under biotic conditions, the performance of the IP-MEDC agreed well with the simulation test, showing a significantly higher chemical oxygen demand (COD) removal rate, desalination rate, and hydrogen production than without the migration pathway. This result is supported by the lowest charge transfer resistance shown by EIS analysis and the abundance of microbes on the bioanode through field emission scanning electron microscopy (FESEM) observation. However, hydrogen production was diminished in the second-fed batch cycle, presumably due to the active diffusion of high Cl¯ concentrations from desalination to the anode chamber, which was detrimental to microbial growth. Enlarging the anode volume by threefold improved the COD removal rate and hydrogen production rate by 1.7- and 3.4-fold, respectively, owing to the dilution effect of Cl¯ in the anode. This implied that the dilution effect satisfies both the microbial viability and conductivity. This study also suggests that the anolyte and catholyte replacement frequencies can be reduced, typically at a prolonged hydraulic retention time, thus minimizing the operating cost (e.g., solution pumping). The use of a high concentration of NaCl (35 g L-1) in the desalination chamber and catholyte provides a condition that is close to practicality.

Air-liquid interface cultivation of Navicula incerta using hollow fiber membranes.

Microalgae cultivation in open ponds requires a large footprint, while most photobioreactors need improvement in the ratio of surface to volume and energy consumption. In this study, polyethersulfone (PES) and poly(vinylidene fluoride) (PVDF) hollow fiber membranes with a large surface area were rearranged into open-ended and dead-ended configurations to improve the air-liquid interface cultivation of Navicula incerta. N. incerta were successfully grown on the porous membrane surface with the nutrients circulating inside the lumen. Fourier-transform infrared spectra showed the accumulation of polysaccharides, proteins and humic acids. Hydrophilic polysaccharides reduced water contact angles on PES and PVDF membranes to 37.2 ± 2.6° and 55.7 ± 3.3°, respectively. However, the porosity of PES (80.1 ± 1.1%) and PVDF (61.3 ± 4.5%) membranes were not significantly affected even after cultivation and harvesting of N. incerta. Scanning electron images further confirmed that N. incerta, cell debris and extracellular organic matter accumulated on the membrane. With large pores and a hydrophobic surface, PVDF hollow fiber membranes offered a greater improvement in N. incerta cell growth rate compared to PES hollow fiber membranes despite using different configurations. In the dead-ended configuration, they even attained the greatest improvement in N. incerta growth rate, up to 54.0%. However, PES hollow fiber membranes only achieved improvement in harvesting efficiency within the range of 18.7-38.0% due to weak cell adhesion. PVDF hollow fiber membranes significantly promoted the growth of microalgae N. incerta through the air-liquid interface system, leading to potential applications in wastewater treatment.

Unveiling antimicrobial activity of microalgae Chlorella sorokiniana (UKM2), Chlorella sp. (UKM8) and Scenedesmus sp. (UKM9).

Microalgae represent promising sources of bioactive compounds for pharmaceutical and industrial applications. The emergence of antibiotic resistant bacteria leads to the need to explore new cost-effective, safe, and potent bioactive compounds from the microalgae. This study aimed to investigate the potential of local microalgae for their antimicrobial properties and bioactive compounds. Three local microalgae namely Chlorella sorokiniana (UKM2), Chlorella sp. UKM8, and Scenedesmus sp. UKM9 biomass methanol extracts (ME) were prepared and tested against Gram-positive and Gram-negative bacteria. Chlorella sp. UKM8-ME showed the highest antibacterial activity. UKM8-ME minimum inhibitory concentrations were in the range of 0.312 to 6.25 mg/mL. Cytotoxicity evaluation using MTT assay showed that the microalgae methanolic extracts did not exhibit cytotoxicity against Vero-cells. The UKM8-ME was mainly containing 28 compounds from the Gas Chromatography-Mass Spectrometry (GC-MS) analysis. Major compounds of UKM8-ME included phenol (18.5%), hexadecanoic acid (18.25%), phytol (14.43%), 9,12-octadecadienoic acid (13.69%), and bicyclo[3.1.1]heptane (7.23%), which have been previously described to possess antimicrobial activity. Hence, Chlorella sp. (UKM8) methanol extracts showed promising antibacterial activity. More comprehensive studies are required to purify these antimicrobial compounds and develop our understanding on their mechanism in UKM8-ME to unleash their specific potential.

Recent advances of natural biopolymeric culture scaffold: synthesis and modification.

Traditionally existing 2D culture scaffold has been inappropriately validated due to the failure in generating the precise therapeutic response. Therefore, this leads to the fabrication of 3D culture scaffold resolving the limitations in the in vivo environment. In recent years, tissue engineering played an important role in the field of bio-medical engineering. Biopolymer material, a novel natural material with excellent properties of nontoxic and biodegradable merits can be served as culture scaffold. This review summarizes the modifications of natural biopolymeric culture scaffold with different crosslinkers and their application. In addition, this review provides the recent progress of natural biopolymeric culture scaffold mainly focusing on their properties, synthesizing and modification and application.

Stability evaluation and formula optimization of cellulose-based scaffold for the air-liquid interface cultivation of Navicula incerta.

Culture scaffolds allow microalgae cultivation with minimum water requirement using the air-liquid interface approach. However, the stability of cellulose-based scaffolds in microalgae cultivation remains questionable. In this study, the stability of regenerated cellulose culture scaffolds was enhanced by adjusting TiO2 loading and casting gap. The membrane scaffolds were synthesized using cellulose dissolved in NaOH/urea aqueous solution with various loading of TiO2 nanoparticles. The TiO2 nanoparticles were embedded into the porous membrane scaffolds as proven by Fourier transform infrared spectra, scanning electron microscopic images, and energy-dispersive X-ray spectra. Although surface hydrophilicity and porosity were enhanced by increasing TiO2 and casting gap, the scaffold pore size was reduced. Cellulose membrane scaffold with 0.05 wt% of TiO2 concentration and thickness of 100 µm attained the highest percentage of Navicula incerta growth rate, up to 37.4%. The membrane scaffolds remained stable in terms of weight, porosity and pore size even they were immersed in acidic solution, hydrogen peroxide or autoclaved at 121 °C for 15 min. The optimal cellulose membrane scaffold is with TiO2 loading of 0.5 wt% and thickness of 100 µm, resulting in supporting the highest N. incerta growth rate and and exhibits good membrane stability.

Sustainable cultivation of Navicula incerta using cellulose-based scaffold incorporated with nanoparticles in air-liquid interface cultivation system.

Microalgae cultivation using open cultivation systems requires large area and it is susceptible to contamination as well as weather changes. Meanwhile, the closed systems require large capital investment, and they are susceptible to the build-up of dissolved oxygen. Air-liquid interface culture systems with low water-footprint, but high packing density can be used for microalgae cultivation if low-cost culture scaffolds are available. In this study, cellulose-based scaffolds were synthesized using NaOH/urea aqueous solution as the solvent. Titanium dioxide (TiO2), silica gel and polyethylene glycol 1000 (PEG 1000) nanoparticles were added into the membrane scaffolds to increase the hydrophilicity of nutrient absorbing to support the growth of microalgae. The membrane scaffolds were characterized by FTIR, SEM, contact angle, porosity and porometry. All three nanoparticles additives showed their ability in reducing the contact angle of membrane scaffolds from 63.4 ± 2.3° to a range of 52.6 ± 1.2° to 38.8 ± 1.5° due to the hydrophilic properties of the nanoparticles. The decreasing in pore size when nanoparticles were added did not affect the porosity of membrane scaffolds. Cellulose membrane scaffold with TiO2 showed the highest percentage of microalgae Navicula incerta growth rate of 22.1% because of the antibacterial properties of TiO2 in lowering the risk of cell contamination and enhancing the growth of N. incerta. The results exhibited that cellulose-based scaffold with TiO2 added could be an effective support in plant cell culture field.

Dose-response analysis of toxic effect from palm oil mill effluent (POME) by-products on biohydrogen producing bacteria - A preliminary study on microbial density and determination of EC50.

The stimulant and toxicity effects of reported organic (acetic acid, propionic acid, butyric acid, formic acid, oil & grease) and inorganic (copper) by-products presented in palm oil mill effluent on anaerobic bacterial population were examined in this paper. The toxicity test had shown that acetic, propionic and butyric acids tend to stimulate the bacterial density level (survival rate more than 50%), while formic acid, copper, oil and grease were shown to have suppressed the density level (survival rate less than 50%). The highest biomass recorded was 1.66 mg/L for the concentration of acetic acid at 216 mg/L and lowest biomass concentration, 0.90 mg/L for copper at 1.40 mg/L. Biohydrogen-producing bacteria have a favourable growth rate around pH 5.5. The comparison of half maximal effective concentration (EC50) values between two test duration on the effects of organic and inorganic by-products postulate that bacteria had a higher tolerance towards volatile fatty acids. While acetic, butyric and propionic acids had exhibited higher tolerance EC50 values for bacteria, but the opposite trend was observed for formic acid, copper and oil & grease.

Enhanced reduction of waste activated sludge at a low temperature by locally isolated strains Pseudomonas sp. VNT and Aeromonas sp. VNT.

Appropriate control of waste activated sludge (WAS) is required to solve the annual increment of WAS volume. The low bacterial activity at low temperatures poses a difficulty in reducing or utilizing WAS in cold weather regions and/or during the winter season. This study reveals a practical method to enhance sludge reduction at a low temperature using isolated strains of Pseudomonas and Aeromonas species. The effect of inoculating each strain into WAS was examined at different temperatures (4°C, 10°C, 12°C, 15°C, 20°C, and 30°C) and under an aerobic condition. Sludge reduction showed 2- to 8-fold improvement at the temperature range from 4°C to 15°C. Both strains are psychrophilic and can produce protease and lipase for sludge degradation even at low temperatures. Thus, biological WAS treatment at a psychrophilic temperature can be enhanced by inoculating these promising strains.

Enhancement of sludge reduction and methane production by removing extracellular polymeric substances from waste activated sludge.

The management of waste activated sludge (WAS) recycling is a concern that affects the development of the future low-carbon society, particularly sludge reduction and biomass utilization. In this study, we investigated the effect of removing extracellular polymeric substances (EPS), which play important roles in the adhesion and flocculation of WAS, on increased sludge disintegration, thereby enhancing sludge reduction and methane production by anaerobic digestion. EPS removal from WAS by ethylenediaminetetraacetic acid (EDTA) significantly enhanced sludge reduction, i.e., 49 ± 5% compared with 27 ± 1% of the control at the end the digestion process. Methane production was also improved in WAS without EPS by 8881 ± 109 CH4 µmol g(-1) dry-weight of sludge. Microbial activity was determined by denaturing gradient gel electrophoresis and real-time polymerase chain reaction, which showed that the hydrolysis and acetogenesis stages were enhanced by pretreatment with 2% EDTA, with a larger methanogenic community and better methane production.