Antiviral Activity of Chlorophyll Extracts from Tetraselmis sp., a Marine Microalga, Against Zika Virus Infection.

Recent advancements in the large-scale cultivation of Tetraselmis sp. in Korea have enabled year-round production of this marine microalgae. This study explores the potential industrial applications of Tetraselmis sp. biomass by investigating the antiviral properties of its extracts and primary components. The antiviral effects of Tetraselmis sp. extracts were evaluated in Zika virus (ZIKV)-infected cells. Following extensive isolation and purification, the main compounds were characterized using liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) analyses. Their antiviral activities were confirmed using in vitro and in silico tests. Tetraselmis sp. extracts reduced infectious viral particles and non-structural protein 1 messenger RNA levels in ZIKV-infected cells without inducing cytotoxicity. Additionally, they modulated the interferon-mediated immune system responses. Tetraselmis sp. extracts are composed of four main chlorophylls: chlorophyll a, chlorin e6-131-152-dimethyl-173-phytyl ester, hydroxychlorophyll a, and hydroxypheophytin a. Among them, chlorophyll a, chlorin e6-131-152-dimethyl-173-phytyl ester, and hydroxypheophytin showed the antiviral activities in ZIKV-infected cells and molecular docking simulations predicted interactions between these chlorophylls and ZIKV. Our findings suggest that Tetraselmis sp. chlorophyll extracts exert antiviral effects against ZIKV and could serve as potential therapeutic candidates against ZIKV infection.

Surfactant-Mediated Microalgal Flocculation: Process Efficiency and Kinetic Modelling.

Microalgae are a valuable source of lipids, proteins, and pigments, but there are challenges in large-scale production, especially in harvesting. Existing methods lack proven efficacy and cost-effectiveness. However, flocculation, an energy-efficient technique, is emerging as a promising solution. Integrating surfactants enhances microalgal harvesting and disruption simultaneously, reducing processing costs. This study investigated cetyltrimethylammonium bromide (CTAB), dodecyltrimethylammonium bromide (DTAB), and sodium dodecyl sulphate (SDS) for harvesting Tetraselmis sp. strains (75LG and 46NLG). CTAB exhibits superior results, with 88% harvesting efficiency at 1500 and 2000 mg L-1 for 75LG and 46NLG, respectively, for 60 min of sedimentation-thus being able to reduce the operating time. Beyond evaluating harvesting efficiency, our study explored the kinetics of the process; the modified Gompertz model led to the best fit. Furthermore, the largest kinetic constants were observed with CTAB, thus highlighting its efficacy in optimising the microalgal harvesting process. With the incorporation of the suggested enhancements, which should be addressed in future work, CTAB could hold the potential to optimise microalgal harvesting for cost-effective and sustainable large-scale production, eventually unlocking the commercial potential of microalgae for biodiesel production.

Harvesting marine microalgae Tetraselmis sp. using cellulose acetate membrane.

This study presents the development and application of a cellulose acetate phase-inversion membrane for the efficient harvesting of Tetraselmis sp., a promising alternative for aquaculture feedstock. Once fabricated, the cellulose acetate membrane was characterized, and its performance was evaluated through the filtration of Tetraselmis sp. broth. The results demonstrated that the developed membrane exhibited exceptional microalgae harvesting efficiency. It showed a low intrinsic resistance and a high clean water permeability of 1100 L/(m2·h·bar), enabling high-throughput filtration of Tetraselmis sp. culture with a permeability of 400 L/(m2·h·bar) and a volume reduction factor of 2.5 ×. The cellulose acetate -based membrane demonstrated robust filtration performance over a 7-day back concentration filtration with minimum irreversible fouling of only 22.5 % irreversibility even without any cleaning. These results highlighted the potential of cellulose acetate as a versatile base polymer for custom-membrane for microalgae harvesting.

Advancing sustainable seawater disinfection: Enhanced inactivation and mechanism of pulsed UV-LEDs irradiation on Tetraselmis sp.

Ultraviolet light-emitting diodes (UV-LEDs), as a novel ultraviolet light source with flexible pulse mode, has gained significant attention for applications in water disinfection and food sterilization. This study investigated the comparative inactivation efficiency of Tetraselmis sp. with continuous and pulsed UV-LEDs irradiation, exploring different wavelengths, duty rates and pulse frequencies. The results reveal a significant enhancement in inactivation efficiency (p < 0.05) under pulsed conditions even at the same UV dose, with inactivation efficiency increasing as duty rate or pulse frequency decreases. The optimal conditions for achieving peak inactivation efficacy are identified as a duty rate of 50% and a pulse frequency of 5 Hz. Within this parameter space, pulsed irradiation leads to a remarkable 1.7-fold increase in inactivation efficiency at UV265 nm and a 1.5-fold increase at UV285 nm compared to continuous irradiation, respectively. Additionally, the disruptive impacts on photosynthetic performance are more pronounced with pulsed irradiation, particularly at the 5 Hz pulse frequency. In shed of these findings, the application of pulsed UV-LEDs irradiation emerges as a promising alternative to the conventional continuous UV disinfection methods in the area of seawater disinfection, offering higher disinfection efficacy and energy consumption.

Cationic Surfactant-Modified Tetraselmis sp. for the Removal of Organic Dyes from Aqueous Solution.

The modification of the Tetraselmis sp. algae material (Tetra-Alg) with surfactant Cethyltrimethylammonium Bromide (CTAB) yielded adsorbent Tetra-Alg-CTAB as an adsorbent of methyl orange (MO) and methylene blue (MB) solutions. The characterization of the adsorbent used an infrared (IR) spectrometer to identify functional groups and Scanning Electron Microscopy with Energy Dispersive X-ray (SEM-EDX FEI Inspect-S50, Midland, ON, Canada) to determine the surface morphology and elemental composition. Methyl orange and methylene blue adsorption on the adsorbent Tetra-Alg, Tetraselmis sp. algae-modified Na+ ions (Tetra-Alg-Na), and Tetra-Alg-CTAB were studied, including variations in pH, contact time, concentration, and reuse of adsorbents. The adsorption of MO and MB by Tetra-Alg-CTAB at pH 10, during a contact time of 90 min, and at a concentration of 250 mg L-1 resulted in MO and MB being absorbed in the amounts of 128.369 and 51.013 mg g-1, respectively. The adsorption kinetics and adsorption isotherms of MO and MB and Tetra-Alg, Tetra-Alg-Na, and Tetra-Alg-CTAB tend to follow pseudo-second-order kinetics models and Freundlich adsorption isotherms with each correlation coefficient value (R2) approaching 1. Due to the modification with the cationic surfactant CTAB, anionic dyes can be strongly sorbed in alkaline pH due to strong electrostatic attraction, while MB is more likely to involve cation exchange and hydrogen bonding. The reuse of Tetra-Alg-CTAB was carried out four times with adsorption percent > 70%, and the adsorbent was very effective in the adsorption of anionic dyes such as MO.

Enhanced Photosynthetic Pigment Production Using a Scaled-Up Continuously Circulated Bioreactor.

Microalgae have gained attention as a promising source of chlorophylls and carotenoids in various industries. However, scaling up of conventional bubble columns presents challenges related to cell sedimentation and the presence of non-photosynthetic cells due to non-circulating zones and decreased light accessibility, respectively. Therefore, this study aimed to evaluate the newly developed continuously circulated bioreactor ROSEMAX at both laboratory and pilot scales, compared to a conventional bubble column. There was no significant difference in the biomass production and photosynthetic pigment content of Tetraselmis sp. cultivated at the laboratory scale (p > 0.05). However, at the pilot scale, the biomass cultured in ROSEMAX showed significantly high biomass (1.69 ± 0.11 g/L, dry weight, DW), chlorophyll-a (14.60 ± 0.76 mg/g, DW), and total carotene (5.64 ± 0.81 mg/g, DW) concentrations compared to the conventional bubble column (1.17 ± 0.11 g/L, DW, 10.67 ± 0.72 mg/g, DW, 3.21 ± 0.56 mg/g, DW, respectively) (p ≤ 0.05). Flow cytometric analyses confirmed that the proportion of Tetraselmis sp. live cells in the culture medium of ROSEMAX was 32.90% higher than that in the conventional bubble column, with a photosynthetic efficiency 1.14 times higher. These results support suggestions to use ROSEMAX as a bioreactor for industrial-scale applications.

Applying an internal transcribed spacer as a single molecular marker to differentiate between Tetraselmis and Chlorella species.

In the realm of applied phycology, algal physiology, and biochemistry publications, the absence of proper identification and documentation of microalgae is a common concern. This poses a significant challenge for non-specialists who struggle to identify numerous eukaryotic microalgae. However, a promising solution lies in employing an appropriate DNA barcoding technique and establishing comprehensive databases of reference sequences. To address this issue, we conducted a study focusing on the molecular characterization and strain identification of Tetraselmis and Chlorella species, utilizing the internal transcribed spacer (ITS) barcode approach. By analyzing the full nuclear ITS region through the Sanger sequencing approach, we obtained ITS barcodes that were subsequently compared with other ITS sequences of various Tetraselmis and Chlorella species. To ensure the reliability of our identification procedure, we conducted a meticulous comparison of the DNA alignment, constructed a phylogenetic tree, and determined the percentage of identical nucleotides. The findings of our study reveal the significant value of the ITS genomic region as a tool for distinguishing and identifying morphologically similar chlorophyta. Moreover, our results demonstrate that both the ITS1 and ITS2 regions are capable of effectively discriminating isolates from one another; however, ITS2 is preferred due to its greater intraspecific variation. These results underscore the indispensability of employing ITS barcoding in microalgae identification, highlighting the limitations of relying solely on morphological characterization.

Antioxidant, Antiviral, and Anti-Inflammatory Activities of Lutein-Enriched Extract of Tetraselmis Species.

Microalgae are proposed to have powerful applications for human health in the pharmaceutical and food industries. Tetraselmis species (sp.), which are green microalgae, were identified as a source of broad-spectrum health-promoting biological activities. However, the bioactivity of these species has not been elucidated. We aimed to confirm the antioxidant, antiviral, and anti-inflammatory effects of Tetraselmis sp. extract (TEE). TEE showed 2,2-diphenyl-1-picryl-hydrazyl-hydrate radical and hydrogen peroxide scavenging activities and reduced plaque formation in Vero E6 cells infected with vaccinia virus. TEE treatment also significantly inhibited nitric oxide (NO) production and improved cell viability in lipopolysaccharide (LPS)-induced RAW264.7 cells. These anti-inflammatory effects were further analyzed in LPS-induced RAW 264.7 cells and the zebrafish model. Further, TEE reduced induced NO synthase expression and proinflammatory cytokine release, including tumor necrosis factor-α, interleukin-6, and interleukin-1ß, through MAPKs and NF-κB-dependent mechanisms. Further analysis revealed that TEE increased the survival rate and reduced cell death and NO production in an LPS-stimulated zebrafish model. Further, high-performance liquid chromatography revealed a strong presence of the carotenoid lutein in TEE. Overall, the results suggest that lutein-enriched TEE may be a potent antioxidant, antiviral, and anti-inflammatory agent that could be sustainably utilized in industrial applications.

Evaluating disinfection performance of ultraviolet light-emitting diodes against the microalga Tetraselmis sp.: Assay methods, inactivation efficiencies, and action spectrum.

Ultraviolet light-emitting diodes (UV-LEDs) are among the most compact devices and safest technologies in water disinfection systems. However, the validation of different assay methods to evaluate the disinfection performance of different wavelengths (265, 280, 285, and 300 nm) of UV-LEDs toward marine microalgae remains poorly characterized. In this study, several detection assays, namely the culture-based most probable number (MPN) assay, membrane integrity-based vital stain (VS) assay, chlorophyll fluorescence assay, and photochemical efficiency assay, were compared to assess the viability of the marine microalga Tetraselmis sp., with results indicating the MPN assay to be the most sensitive. In addition, this study compared the inactivation kinetics, inactivation efficiency, and energy efficiency of Tetraselmis sp. under different UV wavelengths, as assessed by the VS and MPN assays. The fluence-response curves of Tetraselmis sp. varied with assay and wavelength, with Geeraerd's model fitting all fluence-response microalgal inactivation curves. The results showed a non-significant difference in inactivation efficiency among different wavelengths of UV-LEDs (except for 300 nm) when using the VS assay. On the contrary, significant differences among all wavelengths were observed with respect to inactivation efficiency when using the MPN assay. The wavelength of 265 nm exhibited maximum inactivation efficiency, whereas 285 nm achieved optimal energy efficiency. The UV action spectrum of Tetraselmis sp. exhibited the peak at 265 nm, a finding which matched well with the absorbance spectrum of DNA. The observations from this study provide a theoretical basis and technical support for the application of the emerging UV-LED light sources in the algicidal treatment of marine water.

Effect of heavy metals mixture on the growth and physiology of Tetraselmis sp.: Applications to lipid production and bioremediation.

Phycoremediation of heavy metals from contaminated waters by oleaginous microalgae is an eco-friendly and emerging trend. Different concentrations of toxic metals such as nickel (Ni), chromium (Cr) and cobalt (Co) were added in Tetraselmis sp. culture media. Mixture Design was used to model the effect of these metals on cell growth, lipid production and heavy metals removal. Tetraselmis sp. was identified as an outstanding Ni, Cr and Co accumulator with bioconcentration factors of 675.17, 584.9 and 169.81 within binary mixtures (Ni × Cr), (Cr × Co) and (Cr × Co) at 6 mg/L, respectively. Optimization studies showed that the highest cell growth (9.22 × 105 cells/mL), lipid content (31% Dry Weight) and metals removal (91%) were obtained with the optimum binary mixture Ni (54.45 %) and Cr (45.45 %). This work presents interesting results revealing the potential of Tetraselmis sp. for nickel removal up to 97 % combined to its potential for biodiesel production.

Year-Round Cultivation of Tetraselmis sp. for Essential Lipid Production in a Semi-Open Raceway System.

There is increasing demand for essential fatty acids (EFAs) from non-fish sources such as microalgae, which are considered a renewable and sustainable biomass. The open raceway system (ORS) is an affordable system for microalgae biomass cultivation for industrial applications. However, seasonal variations in weather can affect biomass productivity and the quality of microalgal biomass. The aim of this study was to determine the feasibility of year-round Tetraselmis sp. cultivation in a semi-ORS in Korea for biomass and bioactive lipid production. To maximize biomass productivity of Tetraselmis sp., f medium was selected because it resulted in a significantly higher biomass productivity (1.64 ± 0.03 g/L) and lower omega-6/omega-3 ratio (0.52/1) under laboratory conditions than f/2 medium (0.70/1). Then, we used climatic data-based building information modeling technology to construct a pilot plant of six semi-ORSs for controlling culture conditions, each with a culture volume of 40,000 L. Over 1 year, there were no significant variations in monthly biomass productivity, fatty acid composition, or the omega-6/omega-3 ratio; however, the lipid content correlated significantly with photosynthetic photon flux density. During year-round cultivation from November 2014 to October 2017, areal productivity was gradually increased by increasing medium salinity and injecting CO2 gas into the culture medium. Productivity peaked at 44.01 g/m2/d in October 2017. Throughout the trials, there were no significant differences in average lipid content, which was 14.88 ± 1.26%, 14.73 ± 2.44%, 12.81 ± 2.82%, and 13.63 ± 3.42% in 2014, 2015, 2016, and 2017, respectively. Our results demonstrated that high biomass productivity and constant lipid content can be sustainably maintained under Korean climate conditions.

Enhanced lipid productivity in AGP knockout marine microalga Tetraselmis sp. using a DNA-free CRISPR-Cas9 RNP method.

A marine green microalga, Tetraselmis sp., has been studied for the production of biomass and lipids in seawater culture. Since carbohydrate and lipid biosynthesis are competitive metabolic pathways, we attempted to increase lipid synthesis in Tetraselmis by inhibiting carbohydrate synthesis. The main regulatory enzyme in the starch synthesis pathway is ADP-glucose pyrophosphorylase (AGP). AGP loss-of-function mutants were developed using the CRISPR-Cas9 ribonucleoprotein (RNP) delivery system. AGP mutants showed a slight decrease in growth. However, the lipid content in two AGP mutants was significantly enhanced by 2.7 and 3.1 fold (21.1% and 24.1% of DCW), respectively, compared to that in the wild type (7.68% of DCW) under nitrogen starvation. This study is an example of metabolic engineering by genetic editing using the CRISPR-Cas9 RNP method in marine green microalgae. Consequently, starchless Tetraselmis mutants might be considered potential producers of lipids in seawater cultures.

Nutritional Potential and Toxicological Evaluation of Tetraselmis sp. CTP4 Microalgal Biomass Produced in Industrial Photobioreactors.

Commercial production of microalgal biomass for food and feed is a recent worldwide trend. Although it is common to publish nutritional data for microalgae grown at the lab-scale, data about industrial strains cultivated in an industrial setting are scarce in the literature. Thus, here we present the nutritional composition and a microbiological and toxicological evaluation of Tetraselmis sp. CTP4 biomass, cultivated in 100-m3 photobioreactors at an industrial production facility (AlgaFarm). This microalga contained high amounts of protein (31.2 g/100 g), dietary fibres (24.6 g/100 g), digestible carbohydrates (18.1 g/100 g) and ashes (15.2 g/100 g), but low lipid content (7.04 g/100 g). The biomass displayed a balanced amount of essential amino acids, n-3 polyunsaturated fatty acids, and starch-like polysaccharides. Significant levels of chlorophyll (3.5 g/100 g), carotenoids (0.61 g/100 g), and vitamins (e.g., 79.2 mg ascorbic acid /100 g) were also found in the biomass. Conversely, pathogenic bacteria, heavy metals, cyanotoxins, mycotoxins, polycyclic aromatic hydrocarbons, and pesticides were absent. The biomass showed moderate antioxidant activity in several in vitro assays. Taken together, as the biomass produced has a balanced biochemical composition of macronutrients and (pro-)vitamins, lacking any toxic contaminants, these results suggest that this strain can be used for nutritional applications.

The Effects of Microalgae as Live Food for Brachionus plicatilis (Rotifer) in Intensive Culture System.

Brachionus plicatilis is used to feed fish and crustacean larvae in the aquaculture industry. It is well established that the type of microalgae may influence rotifer production. This experiment was conducted to determine the effect of five different locally available microalgae species at Fisheries Research Institute (FRI), Kampung Pulau Sayak, Kedah, Malaysia on the instantaneous growth rate (µ) of rotifer. Nannochloris sp., Tetraselmis sp., Isochrysis sp., Chlorella sp., and Nannochloropsis sp. were used as feed at different algae densities (0.1, 0.3, 0.7 and 1.5 × 106 cells/ml) and culture volumes (20, 70 and 210 ml). At algae densities ranging from 0.3 to 1.5 × 106 cells/ml, an average µ value of more than 0.90 per day were recorded for all algae species. However, at density of 0.1 × 106 cells/ml, only Tetraselmis sp. resulted in the significantly highest µ value compared with others (p < 0.05). In terms of volume, smaller culture volume of Tetraselmis sp. (20 ml) showed significantly higher µ compared with higher volume (70 and 210 ml cultures).

Flocculation Effect of Alkaline Electrolyzed Water (AEW) on Harvesting of Marine Microalga Tetraselmis sp.

Microalgae hold promise as a renewable energy source for the production of biofuel, as they can convert light energy into chemical energy through photosynthesis. However, cost-efficient harvest of microalgae remains a major challenge to commercial-scale algal biofuel production. We first investigated the potential of electrolytic water as a flocculant for harvesting Tetraselmis sp. Alkaline electrolyzed water (AEW) is produced at the cathode through water electrolysis. It contains mineral ions such as Na+, K+, Ca2+, and Mg2+ that can act as flocculants. The flocculation activity with AEW was evaluated via culture density, AEW concentration, medium pH, settling time, and ionic strength analyses. The flocculation efficiency was 88.7% at 20% AEW (pH 8, 10 min) with a biomass concentration of 2 g/l. The initial biomass concentration and medium pH had significant influences on the flocculation activity of AEW. A viability test of flocculated microalgal cells was conducted using Evans blue stain, and the cells appeared intact. Furthermore, the growth rate of Tetraselmis sp. in recycled flocculation medium was similar to the growth rate in fresh F/2 medium. Our results suggested that AEW flocculation could be a very useful and affordable methodology for fresh biomass harvesting with environmentally friendly easy operation in part of the algal biofuel production process.

Enhanced Production of Fatty Acids via Redirection of Carbon Flux in Marine Microalga Tetraselmis sp.

Lipids in microalgae are energy-rich compounds and considered as an attractive feedstock for biodiesel production. To redirect carbon flux from competing pathways to the fatty acid synthesis pathway of Tetraselmis sp., we used three types of chemical inhibitors that can block the starch synthesis pathway or photorespiration, under nitrogen-sufficient and nitrogen-deficient conditions. The starch synthesis pathway in chloroplasts and the cytosol can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 1,2-cyclohexane diamine tetraacetic acid (CDTA), respectively. Degradation of glycine into ammonia during photorespiration was blocked by aminooxyacetate (AOA) to maintain biomass concentration. Inhibition of starch synthesis pathways in the cytosol by CDTA increased fatty acid productivity by 27% under nitrogen deficiency, whereas the blocking of photorespiration in mitochondria by AOA was increased by 35% under nitrogen-sufficient conditions. The results of this study indicate that blocking starch or photorespiration pathways may redirect the carbon flux to fatty acid synthesis.

A general reaction network and kinetic model of the hydrothermal liquefaction of microalgae Tetraselmis sp.

In this work, the hydrothermal liquefaction (HTL) of microalgal Tetraselmis sp. was conducted at various reaction temperatures (250-350°C) and reaction times (10-60min). A general reaction network and a quantitative kinetic model were proposed for the HTL of microalgae. In this reaction network, the primary decomposition of lipids, proteins, and carbohydrates generated heavy oil (HO), light oil (LO), and aqueous-phase (AP) products. Then, reversible interconversions and further decomposition of these product fractions to produce gas product were followed. The model accurately captures the trends observed in the experimental data. Analyses of the kinetic parameters (reaction rate constants and activation energies) suggested the dominant reaction pathways as well as the contribution of the biochemical compositions to the bio-oil yield. Finally, the kinetic parameters calculated from the model were utilized to explore the parameter space in order to predict the liquefaction product yields depending on the reaction time and temperature.

Effects of nutritional conditions on growth and biochemical composition of Tetraselmis sp.

BACKGROUND: This study aimed to maximize biomass concentration, biomass productivity and biochemical composition of the marine microalga Tetraselmis sp. METHODS: In the current study, Box-Behnken Design was used to model the effect of NaNO3, NaH2PO4, metals and vitamins in the F/2 medium on the growth, total chlorophylls, carotenoids and starch contents. The total chlorophylls content was quantified by spectrophotometry. The FT-IR spectroscopy was used to estimate the biochemical compositions of Tetraselmis sp. grown under both optimized medium culture for starch production and standard culture medium. RESULTS: Finalized NaNO3 (1.76 mM), NaH2PO4 (0.018 mM), metals (1500 µL.L-1) and vitamins (312.5 µL.L-1) concentrations, generated an increase in biomass concentration up to 5.72 g.L-1 which contributed to an increase about 2.4-fold than that of the standard conditions of biomass productivity (408.57 mg.L-1.day-1). The maximum value of carotenoids content (0.3 mg.g DW-1) was achieved at the highest level of all factors. The total chlorophylls content reached also its maximum (5.18 mg.g DW-1) at high nitrate (1.76 mM), phosphate (0.054 mM), metals and vitamins concentrations, while the maximum starch content (42% DW) was achieved with low nitrate and phosphate concentrations (0.58 mM and 0.027 mM) and with metals and vitamins limitations. Thus, the nitrogen, phosphorus, metals and vitamins limitations led to divert the metabolism for the starch biosynthesis. CONCLUSIONS: The high biomass concentration productivity and starch production make Tetraselmis sp. strain a good candidate for biotechnological applications.

Urban wastewater treatment by Tetraselmis sp. CTP4 (Chlorophyta).

The ability of a recent isolate, Tetraselmis sp. CTP4, for nutrient removal from sewage effluents before and after the nitrification process under batch and continuous cultivation was studied. Biomass productivities in both wastewaters were similar under continuous conditions (0.343±0.053gL-1d-1) and nutrient uptake rates were maximal 31.4±0.4mgNL-1d-1 and 6.66±1.57mgP-PO43-L-1d-1 in WW before nitrification when cultivated in batch. Among batch treatments, cellular protein, carbohydrate and lipid levels shifted with aging cultures from 71.7±6.3 to 29.2±1.2%, 17.4±7.2 to 57.2±3.9% and 10.9±1.7 to 13.7±4.7%, respectively. In contrast, CTP4 cultivated continuously in Algal medium (control) showed lower biomass productivities (0.282gVSSL-1d-1) although improved lipid content (up to 20% lipids) in batch cultivation. Overall, Tetraselmis sp. CTP4 is promising for WW treatment as a replacement of the costly nitrification process, fixating more nutrients and providing a protein and carbohydrate-rich biomass as by-product.

Enhanced lipid and biomass production by a newly isolated and identified marine microalga.

BACKGROUND: The increasing demand for microalgae lipids as an alternative to fish has encouraged researchers to explore oleaginous microalgae for food uses. In this context, optimization of growth and lipid production by the marine oleaginous V2-strain-microalgae is of great interest as it contains large amounts of mono-unsaturated (MUFAs) and poly-unsaturated fatty acids (PUFAs). METHODS: In this study, the isolated V2 strain was identified based on 23S rRNA gene. Growth and lipid production conditions were optimized by using the response surface methodology in order to maximize its cell growth and lipid content that was quantified by both flow cytometry and the gravimetric method. The intracellular lipid bodies were detected after staining with Nile red by epifluorescence microscopy. The fatty acid profile of optimal culture conditions was determined by gas chromatography coupled to a flame ionization detector. RESULTS: The phenotypic and phylogenetic analyses showed that the strain V2 was affiliated to Tetraselmis genus. The marine microalga is known as an interesting oleaginous species according to its high lipid production and its fatty acid composition. The optimization process showed that maximum cell abundance was achieved under the following conditions: pH: 7, salinity: 30 and photosynthetic light intensity (PAR): 133 µmol photons.m-2.s-1. In addition, the highest lipid content (49 ± 2.1% dry weight) was obtained at pH: 7, salinity: 37.23 and photosynthetic light intensity (PAR): 188 µmol photons.m-2.s-1. The fatty acid profile revealed the presence of 39.2% and 16.1% of total fatty acids of mono-unsaturated fatty acids (MUFAs) and poly-unsaturated fatty acids (PUFAs), respectively. Omega 3 (ω3), omega 6 (ω6) and omega 9 (ω9) represented 5.28%, 8.12% and 32.8% of total fatty acids, respectively. CONCLUSIONS: This study showed the successful optimization of salinity, light intensity and pH for highest growth, lipid production and a good fatty acid composition, making strain V2 highly suitable for food and nutraceutical applications.