Growing microalgae as aquaculture feeds on twin-layers: a novel solid-state photobioreactor

Four strains of marine microalgae commonly used as live feeds in hatcheries (Isochrysis sp. T.ISO, Tetraselmis suecica, Phaeodactylum tricornutum, Nannochloropsis sp.) were grown in a novel solid-state photobioreactor, the twin-layer system. Microalgae were immobilized by self adhesion to vertically oriented twin-layer modules which consisted of two different types of ultrathin layers, a macroporous source layer (glass fiber nonwoven) through which the culture medium was transported by gravity flow, and a microporous substrate layer (plain printing paper) which carried the algae on both surfaces of the source layer. This simple open cultivation system effectively separated the immobilized microalgae from the bulk of the growth medium and permitted prolonged cultivation of microalgae with average biomass yields of 10–15 g dry weight m^?2 growth area after 14–25 days of cultivation. Algal biomass was harvested as fresh weight (with 72–84 % water content) without the need to pre-concentrate algae. No aeration or external CO₂ supply was necessary, and due to the microporous substrate layer, no eukaryotic contaminations were observed during the experiment. All experiments were conducted in Germany under greenhouse conditions with natural sunlight. Small-scale growth experiments performed under the same conditions revealed that growth over most of the experimental period (24 days) was linear in all tested algae with growth rates (dry weight per square meter growth area) determined to be 0.6 g ?m^?2?day^?1 (Isochrysis), 0.8 g? m^?2?day^?1 (Nannochloropsis), 1.5 g ?m^?2?day^?1 (Tetraselmis), and 1.8 g? m^?2?day^?1 (Phaeodactylum). Due to its cost-effective construction and with further optimisation of design and productivity at technical scales, the twin-layer system may provide an attractive alternative to methods traditionally used to cultivate live microalgae.     

Mixotrophic and photoautotrophic cultivation of 14 microalgae isolates from Saskatchewan, Canada: potential applications for wastewater remediation for biofuel production

Northern regions are generally viewed as unsuitable for microalgal biofuel production due to unfavorable climate and solar insolation levels. However, these conditions can potentially be mitigated by coupling microalgal cultivation to industrial processes such as wastewater treatment. In this study, we have examined the biomass and lipid productivity characteristics of 14 microalgae isolates (Chlorophyta) from the Canadian province of Saskatchewan. Under both photoautotrophic and mixotrophic cultivation, a distinct linear trend was observed between biomass and lipid productivities in the 14 SK isolates. The most productive strain under cultivation in TAP media was Scenedesmus sp.-AMDD which displayed rates of biomass and fatty acid productivities of 80 and 30.7?mg?L^?1?day^?1, respectively. The most productive strain in B3NV media was Chlamydomonas debaryana-AMLs1b which displayed rates of biomass and fatty acid productivities of 51.7 and 5.9?mg?L^?1?day^?1, respectively. In 11 of the isolates tested, secondary municipal wastewater (MCWW) supported rates of biomass productivity between 21 and 33?mg?L^?1?day^?1 with Scenedesmus sp.-AMDD being the most productive. Three strains, Chlamydomonas debaryana-AMB1, Chlorella sorokiniana-RBD8 and Micractinium sp.-RB1b, showed large increases in biomass productivity when cultivated mixotrophically in MCWW supplemented with glycerol. High relative oleic acid content was detected in 10 of the 14 isolates when grown mixotrophically in media supplemented with acetate. There was no detectable effect on the fatty acid profiles in cells cultivated mixotrophically in glycerol-supplemented MCWW. These data indicate that biomass and lipid productivities are boosted by mixotrophic cultivation. Exploiting this response in municipal wastewater is a promising strategy for the production of environmentally sustainable biofuels.     

Continuous microalgae cultivation in a photobioreactor

New biomass sources for alternative fuels has become a subject of increasing importance as the nation strives to resolve the economic and strategic impacts of limited fossil fuel resources on our national security, environment, and global climate. Algae are among the most promising non‐food‐crop‐based biomass feedstocks. However, there are currently no commercially viable microalgae‐based production systems for biofuel production that have been developed, as limitations include less‐than optimal oil content, growth rates, and cultivation techniques. While batch studies are critical for determining basic growth phases and characteristics of the algal species, steady‐state studies are necessary to better understand and measure the specific growth parameters. This study evaluated the effects of dilution rate on microalgal biomass productivity, lipid content, and fatty acid profile under steady‐state conditions with continuous illumination and carbon dioxide supplemention for two types of algae. Continuous cultures were conducted for more that 3 months. Our results show that the productivity of Chlorella minutissima varied from 39 to 137 mg/L/day (dry mass) when the dilution rate varied from 0.08 to 0.64 day^?1. The biomass productivity of C. minutissima reached a maximum value (137 mg/L/day) at a dilution rate of 0.33 day^?1, while the productivity of Dunaliella tertiolecta varied from 46 to 91 mg/L/day at a dilution rate of 0.17 to 0.74 day^?1. The biomass productivity of D. tertiolecta reached a maximum value of 91 mg/L/day at a dilution rate of 0.42 day^?1. Moreover, the lipid content had no significant change with various dilution rates. Biotechnol. Bioeng. 2012; 109: 2468–2474. © 2012 Wiley Periodicals, Inc.     

Enhancement of total lipid yield by nitrogen,carbon, and iron supplementation in isolated microalgae

The biochemical contents and biodiesel production ability of three microalgal strains grown under different sodium nitrate, sodium carbonate, and ferric ammonium citrate (iron) levels were investigated. The highest biomass and lipid contents were found in Scenedesmus sp., Chlorella sp., and Chlamydomonas sp. when grown in normal BG‐11 containing sodium carbonate concentration at 0.03 g · L^?1, and in normal BG‐11 containing iron concentration (IC) at 0.009 or 0.012 g · L^?1. Increasing the sodium nitrate level increased the biomass content, but decreased the lipid content in all three microalgae. Among the three microalgae, Scenedesmus sp. showed the highest total lipid yield of 0.69 g · L^?1 under the IC of 0.012 g · L^?1. Palmitic and oleic acids were the major fatty acids of Scenedesmus sp. and Chlamydomonas sp. lipids. On the other hand, Chlorella sp. lipids were rich in palmitic, oleic, and linolenic acids, and henceforth contributing to poor biodiesel properties below the standard limits. The three isolated strains had a potential for biodiesel production. Nevertheless, Scenedesmus sp. from stone quarry pond water was the most suitable source for biodiesel production with tolerance toward the high concentration of sodium carbonate without the loss of its biodiesel properties.     

Comparison of growth of Tetraselmis in a tubular photobioreactor (Biocoil) and a raceway pond

Microalgae cultivation systems can be divided broadly into open ponds and closed photobioreactors. This study investigated the growth and biomass productivity of the halophilic green alga Tetraselmis sp. MUR-233, grown outdoors in paddle wheel-driven open raceway ponds and in a tubular closed photobioreactor (Biocoil) at a salinity of 7 % NaCl (w/v) between mid-March and June 2010 (austral autumn/winter). Volumetric productivity in the Biocoil averaged 67 mg ash-free dry weight (AFDW) L^?1 day^?1 when the culture was grown without CO₂ addition. This productivity was 86 % greater, although less stable, than that achieved in the open raceway pond (36 mg L^?1 day^?1) grown at the same time in the autumn period. The Tetraselmis culture in the open raceway pond could be maintained in semi-continuous culture for the whole experimental period of 3 months without an additional CO₂ supply, whereas in the Biocoil, under the same conditions, reliable semi-continuous culture was only achievable for a period of 38 days. However, stable semi-continuous culture was achieved in the Biocoil by the addition of CO₂ at a controlled pH of ~7.5. With CO₂ addition, the volumetric biomass productivity in the Biocoil was 85 mg AFDW L^?1 day^?1 which was 5.5 times higher than the productivity achieved in the open raceway pond (15 mg AFDW L^?1 day^?1) with CO₂ addition and 8 times higher compared to the productivity in the open raceway pond without CO₂ addition (11 mg AFDW L^?1 day^?1), when cultures were grown in winter. The illuminated area productivities highlight an alternative story and showed that the open raceway pond had a three times higher productivity (3,000 mg AFDW m^?2 day^?1) compared to the Biocoil (850 mg AFDW m^?2 day^?1). Although significant differences were found between treatments and cultivation systems, the overall average lipid content for Tetraselmis sp. MUR-233 was 50 % in exponential phase during semi-continuous cultivation.     

Evaluation of halophilic microalgae isolated from Rabigh Red Sea coastal area for biodiesel production: Screening and biochemical studies

In the present study, different water samples from Red Sea coastal area at Rabigh city, Saudi Arabia were studied for their dominant algal species. Microalgal isolation was carried out based on dilution method and morphologically examined using F/2 as a growth medium. Dry weight and main biochemical composition (protein, carbohydrates, lipids) of all species were performed at the end of the growth, and biodiesel characteristics were estimated. Nannochloropsis sp., Dunaliella sp., Tetraselmis sp., Prorocentrum sp., Chlorella sp., Nitzschia sp., Coscinodiscus sp., and Navicula sp. were the most dominant species in the collected water samples and were used for further evaluation. Nannochloropsis sp. surpassed all other isolates in concern of biomass production with the maximum recorded dry weight of 0.89 g L^?1, followed by Dunaliella sp. (0.69 g L^?1). The highest crude protein content was observed in Nitzschia sp. (38.21%) and Dunaliella sp. (18.01%), while Nannochloropsis sp. showed 13.38%, with the lowest recorded lipid content in Coscinodiscus sp. (10.09%). Based on the growth, lipid content, and biodiesel characteristics, the present study suggested Dunaliella sp. and Nitzschia sp. as promising candidates for further large-scale biodiesel production.     

Evaluation of indigenous microalgal isolate Chlorella sp. FC2 IITG as a cell factory for biodiesel production and scale up in outdoor conditions

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

The effect of gradual increase in salinity on the biomass productivity and biochemical composition of several marine,halotolerant<Emphasis Type="Bold">,</Emphasis> and halophilic microalgae

Open ponds are the preferred cultivation system for large-scale microalgal biomass production. To be more sustainable, commercial scale biomass production should rely on seawater, as freshwater is a limiting resource, especially in places with high irradiance. If seawater is used for both pond fill and evaporative volume makeup, salinity of the growth media will rise over time. It is not possible for any species to achieve optimum growth over the whole saline spectrum (from seawater salinity level up to salt saturation state). In this study, we investigated the effects of gradual salinity increase (between 35 and 233 ppt) on biomass productivity and biochemical composition (lipid and carbohydrate) of six marine, two halotolerant, and a halophilic microalgae. A gradual and slow stepped salinity increase was found to expand the salinity tolerance range of tested species. A gradual reduction in biomass productivity and maximum photochemical efficiency was observed as a consequence of increased salinity in all tested species. Among the marine microalgae, Tetraselmis showed highest biomass productivity (32 mg L^?1 day^?1) with widest salinity tolerance range (35 to 109 ppt). Halotolerant Amphora and Navicula were able to grow from 35 ppt to 129 ppt salinity. Halophilic Dunaliella was the only species capable of growing between 35 and 233 ppt and showed highest lipid content (56.2%) among all tested species. This study showed that it should be possible to maintain high biomass in open outdoor cultivation utilizing seawater by growing Tetraselmis, Amphora, and Dunaliella one after another as salinity increases in the cultivation system.     

Assessing near-infrared reflectance spectroscopy for the rapid detection of lipid and biomass in microalgae cultures

With intensification of interest in microalgae as a source of biomass for biofuel production, rapid methods are needed for lipid screening of cultures. In this study, near-infrared reflectance spectroscopy (NIRS) was assessed as a method for analysing lipid (specifically, total fatty acid methyl esters (FAME) obtainable from processing) and biomass in late logarithmic and stationary phase cultures of the green alga Kirchneriella sp. and the eustigmatophyte Nannochloropsis sp. Culture samples were filtered, scanned by NIRS and chemically analysed; by combining these sets of information, models were developed to predict total biomass, FAME content and FAME as a percentage of dry weight in samples. Chemically derived (actual) and NIRS-predicted data were compared using the coefficient of determination (R ²) and the ratio of the standard deviation (SD) of actual data to the SD of NIRS prediction (RPD). For Kirchneriella sp. samples, models gave excellent prediction (R ²?≥?0.96; RPD?≥?4.8) for all parameters. For Nannochloropsis sp., the model metrics were less favourable (R ²?=?0.84–0.94; RPD?=?2.5–4.2), though sufficient to provide estimations that could be useful for screening purposes. This technique may require further validation and comparison with other species, but this study shows the potential of the NIRS as a rapid screening method (e.g. up to 200 sample analyses per day) for estimating FAME or other microalgal constituents and encourages further investigation.     

Improvement of biomass and fatty acid productivity in ocean cultivation of <Emphasis Type="Italic">Tetraselmis</Emphasis> sp. using hypersaline medium

In this study, hypersaline media were used for ocean cultivation of the marine microalga Tetraselmis sp. KCTC12432BP for enhanced biomass and fatty acid (FA) productivity. Hypersaline media (55, 80, and 105 PSU) were prepared without sterilization by addition of NaCl to seawater obtained from Incheon, Korea. The highest biomass productivity was obtained at 55 PSU (0.16 g L^?1 day^?1) followed by 80 PSU (0.15 g L^?1 day^?1). Although the specific growth rate of Tetraselmis decreased at salinities higher than 55 PSU, prevention of contamination led to higher biomass productivity at 80 PSU than at 30 PSU (0.03 g L^?1 day^?1). FA content of algal biomass increased as salinity increased to 80 PSU, above which it declined, and FA productivity was highest at 80 PSU. Ocean cultivation of Tetraselmis was performed using 50-L tubular module photobioreactors and 2.5-kL square basic ponds, closed- and open-type ocean culture systems, respectively. Culturing microalgae in hypersaline medium (80 PSU) improved biomass productivities by 89 and 152% in closed and open cultures, respectively, compared with cultures with regular salinity. FA productivity was greatly improved by 369% in the closed cultures. The efficacy of salinity shift and N-deficiency to enhance FA productivity was also investigated. Lowering salinity to 30 PSU with N-starvation following cultivation at 80 PSU improved FA productivity by 19% in comparison with single-stage culture without N-deficiency at 30 PSU. The results show that salinity manipulation could be an effective strategy to improve biomass and FA productivity in ocean cultivation of Tetraselmis sp.     

The effect of temperature on growth and lipid and fatty acid composition on marine microalgae used for biodiesel production

Cultivation temperature is one of the major factors affecting the growth and lipid accumulation of microalgae. In this study, the effects of temperature on the growth, lipid content, fatty acid composition and biodiesel properties of the marine microalgae Chaetoceros sp. FIKU035, Tetraselmis suecica FIKU032 and Nannochloropsis sp. FIKU036 were investigated. These species were cultured at different temperatures (25, 30, 35 and 40 °C). The results showed that the specific growth rate, biomass and lipid content of all microalgae decreased with increasing temperature. With regards to fatty acids, the presence of saturated fatty acids (SFAs) in T. suecica FIKU032 and Nannochloropsis sp. FIKU036 decreased with increasing temperature, in contrast with polyunsaturated fatty acids (PUFAs). Moreover, Chaetoceros sp. FIKU035 was the only species that could grow at 40 °C. The highest lipid productivity was observed in Chaetoceros sp. FIKU035 when cultivated at 25 °C (66.73 ± 1.34 mg L^?1 day^?1) and 30 °C (61.35 ± 2.89 mg L^?1 day^?1). Moreover, the biodiesel properties (cetane number, cold filter plugging point, kinematic viscosity and density) of the lipids obtained from this species were in accordance with biodiesel standards. This study indicated that Chaetoceros sp. FIKU035 can be considered as a suitable species for biodiesel production in outdoor cultivation.     

Improvement of both lipid and biomass productivities of Qatar Chlorocystis isolate for biodiesel production and food security

Microalgae are considered a very promising alternative for biofuel production. Several strategies were developed to modulate and improve algae metabolites production to meet the requirements for biodiesel production. Most previous research evidenced that the increase of the lipid content is accompanied by a decrease of the biomass production, which increases the cost of the downstream processing. Hence, the challenge is to find special culture conditions that increase the lipid and the biomass productivities simultaneously. In the present work, we developed a strategy for the improvement of biomass and lipid productivities in a novel local microalga isolate, Chlorocystis sp. QUCCCM14, which was not previously known as a promising strain. Indeed, culturing QUCCCM14 using f/2 medium with 10× NaH₂PO₄ (0.15 g L^?1 NaNO₃ and 5.6 mg L^?1 NaH₂PO₄) resulted in an improvement of 3.178 folds the lipid productivity reaching 56.121 mg L^?1 day^?1 and enhanced the biomass productivity reaching 141.363 mg L^?1 day^?1, simultaneously. Comparative analyses of the FAME profiles demonstrated that fed‐batch culture with phosphate or nitrate separately leads to a high production of the omega 3 fatty acids (Linolenic acid), whereas fed‐batch culture with phosphate and nitrate simultaneously increased the production of fatty acids suitable for biodiesel production.     

Microalgae for phosphorus removal and biomass production: a six species screen for dual‐purpose organisms

Microalgae biofuel production can be feasible when a second function is added, such as wastewater treatment. Microalgae differ in uptake of phosphorus (P) and growth, making top performer identification fundamental. The objective of this screen was to identify dual‐purpose candidates capable of high rates of P removal and growth. Three freshwater – Chlorella sp., Monoraphidium minutum sp., and Scenedesmus sp. – and three marine – Nannochloropsis sp., N. limnetica sp., and Tetraselmis suecica sp. – species were batch cultured in 250 mL flasks over 16 days to quantitate total phosphorus (TP) removal and growth as a function of P loads (control, and 5, 10, and 15 mg L^?1 enrichment of control). Experimental design used 100 μmol m^?2 s^?1 of light, a light/dark cycle of 14/10 h, and no CO₂ enrichment. Phosphorus uptake was dependent on species, duration of exposure, and treatment, with significant interaction effects. Growth was dependant on species and treatment. Not all species showed increased P removal with increasing P addition, and no species demonstrated higher growth. Nannochloropsis sp and N. limnetica sp. performed poorly across all treatments. Two dual‐purpose candidates were identified. At the 10 mg L^?1 treatment Monoraphidium minutum sp. removed 67.1% (6.66 mg L^?1 ± 0.60 SE) of TP at day 8, 79.3% (7.86 mg L^?1 ± 0.28 SE) at day 16, and biomass accumulation of 0.63 g L^?1 ± 0.06 SE at day 16. At the same treatment Tetraselmis suecica sp. removed 79.4% (6.98 mg L^?1 ± 0.24 SE) TP at day 8, 83.0% (7.30 mg L^?1 ± 0.60 SE) at day 16, and biomass of 0.55 g L^?1 ± 0.02 SE at day 16. These species merit further study using high‐density wastewater cultures and lipid profiling to assess suitability for a nutrient removal and biomass/biofuel production scheme.     

Process optimization and modeling for the cultivation of Nannochloropsis sp. and Tetraselmis striata via response surface methodology

The aim of this study was to determine the optimal physical process conditions for the cultivation of locally isolated strains of Nannochloropsis sp. and Tetraselmis striata to achieve maximum growth rate. It was essential to evaluate biomass production at different agitation rates, light intensities, and temperature levels. Central composite design and response surface methodology were applied to design the experiments and optimize the cultivation process for Nannochloropsis sp. and T. striata. The specific growth rate of 0.250 d^?1 was obtained for Nannochloropsis sp. cells under the light intensity of 54 μmol photons · m^?2 · s^?1, at the agitation rate of 151 rpm in 24.5°C. The optimal physical process conditions for T. striata were obtained under the light intensity of 56 μmol photons · m^?2 · s^?1 in 25.5°C at the agitation rate of 151 rpm in 25.5°C, resulting in a specific growth rate of 0.226 d^?1. The predicted values were justified by the verification tests. Good agreement between the predicted values and the experimental values confirmed the validity of the models for the cultivation of microalgal strains. In this article, the noteworthy result was that temperature was a dominant factor in obtaining high chl‐a content for Nannochloropsis sp., whereas the growth of T. striata strongly depended on light exposure.     

Interspecific biodiversity enhances biomass and lipid productivity of microalgae as biofuel feedstock

Large improvements in biomass and lipid production are required to make massive scale algal biodiesel production an economic reality. The application of the biodiversity strategy to enhance algal biomass as biofuel feedstock is little. The algal diversity was manipulated in this study to investigate the effects of a combination of biodiversity complementarity and a new medium consisting of seawater and agricultural fertilizer on lipid productivity. The algae diverse community includes two strains of Dunaliella salina (Dunaliella salina 19/30 and 19/18) and three species of Nannochloropsis (Nannochloropsis oculata, Nannochloropsis salina, and Nannochloropsis gaditana). The results showed that the most diverse community (5 species) produced an average of sixfold more biomass in the new medium than did the standard f/2 culture medium. The most diverse polyculture had the highest growth rate (1.01 day^?1), biomass (1.2 g L^?1), and lipid productivity (0.31 g L^?1 day^?1). The assessment of algal polycultures relative to monocultures is particularly interesting and novel for this biofuel field, and the observations that these polycultures resulted in significant lipid productivity improvements are very useful addition to the biofuel research. The possible mechanism (resource diversity) to explain the synergy in mixed cultures warrants further investigation.     

A quantitative analysis of microalgal lipids for optimization of biodiesel and omega‐3 production

The lipid characteristics of microalgae are known to differ between species and change with growth conditions. This work provides a methodology for lipid characterization that enables selection of the optimal strain, cultivation conditions, and processing pathway for commercial biodiesel production from microalgae. Two different microalgal species, Nannochloropsis sp. and Chlorella sp., were cultivated under both nitrogen replete and nitrogen depleted conditions. Lipids were extracted and fractionated into three major classes and quantified gravimetrically. The fatty acid profile of each fraction was analyzed using GC–MS. The resulting quantitative lipid data for each of the cultures is discussed in the context of biodiesel and omega‐3 production. This approach illustrates how the growth conditions greatly affect the distribution of fatty acid present in the major lipid classes and therefore the suitability of the lipid extracts for biodiesel and other secondary products. Biotechnol. Bioeng. 2013; 110: 2096–2104. © 2013 Wiley Periodicals, Inc.     

Biomass and Lipid Production of Dinoflagellates and Raphidophytes in Indoor and Outdoor Photobioreactors

The principal fatty acids from the lipid profiles of two autochthonous dinoflagellates (Alexandrium minutum and Karlodinium veneficum) and one raphidophyte (Heterosigma akashiwo) maintained in bubble column photobioreactors under outdoor culture conditions are described for the first time. The biomass production, lipid content and lipid productivity of these three species were determined and the results compared to those obtained when the strains were cultured indoors. Under the latter condition, the biotic values did not significantly differ among species, whereas under outdoor conditions, differences in both duplication time and fatty acids content were observed. Specifically, A. minutum had higher biomass productivity (0.35 g·L^?1 day^?1), lipid productivity (80.7 mg lipid·L^?1 day^?1) and lipid concentration (252 mg lipid·L^?1) at harvest time (stationary phase) in outdoor conditions. In all three strains, the growth rate and physiological response to the light and temperature fluctuations of outdoor conditions greatly impacted the production parameters. Nonetheless, the species could be successfully grown in an outdoor photobioreactor and were of sufficient robustness to enable the establishment of long-term cultures yielding consistent biomass and lipid production.     

Biomass and lipid enhancement in Ankistrodesmus sp. cultured with reused and minimal nutrients media

Microalgae are a promising feedstock for biofuel production. Lipid content in microalgae could be enhanced under nutrient depletion. This work investigated the effect of the nutrient on lipid accumulation in Ankistrodesmus sp. culture. Batch cultures were carried out using fresh BG11 medium, and after the harvest, the medium was reused for the next culture; this method was repeated two times. The maximum lipid productivity of 29.75 mg L^?1 day^?1 was obtained from the culture with the second reuse medium. In continuous cultures, Ankistrodesmus sp. was cultured in both fresh and modified BG11 mediums. The modified BG11 medium was adjusted to resemble the content of the first reuse medium. As a comparison between batch and continuous cultures, it was proven that the productivity in the continuous culture was better than in the batch, where the achievable maximum biomass and lipid were 188.30 and 38.32 mg L^?1 day^?1. The maximum lipid content of 34.22% was obtained from the continuous culture at a dilution rate of 0.08 day^?1, whereas the maximum saturated and unsaturated fatty acid productivities of 79.96 and 104.54 mg L^?1 day^?1 were obtained at a dilution rate of 0.16 day^?1.     

Effects of initial population density (IPD) on growth and lipid composition of Nannochloropsis sp.

The microalga Nannochloropsis sp. was cultured under different initial population densities (IPDs) ranging from 0.11 to 9.09 g L^?1. The IPD affected the biomass and lipid accumulation significantly. The algal cultured with higher IPD resulted higher biomass concentration (up to 13.07 g L^?1) in 10 days growth. The biomass productivity with 0.98 g L^?1 IPD was 0.75 g L^?1 d^?1 which was higher than that of other IPDs. For IPDs ranging from 0.11 to 0.98 g L^?1, with the increase of IPD, the biomass productivity increased, while for IPD over 0.98 g L^?1, the biomass productivity decreased. Lipid content of the algal culture started with 0.11 g L^?1 IPD reached to 42 % of dry weight. But with the increase of IPD, the lipid content decreased. Lipid composition was analyzed using thin layer chromatography coupled with flame ionization detection (TLC/FID). Seven lipid classes were identified and quantified. The main reserve lipid, triacylglyceride (TAG), accumulated under all different IPD conditions. However, with the increasing IPD values, TAG content decreased from 59.1 to 23.5 % of the total lipids. Based on these results, to obtain the maximal biomass productivity and lipid productivity of Nannochloropsis sp. in mass cultivation systems, it is necessary to select an appropriate IPD.     

Effects of nitrogen sources on cell growth and lipid accumulation of green alga <Emphasis Type="Italic">Neochloris oleoabundans</Emphasis>

Microalgal lipids are the oils of future for sustainable biodiesel production. However, relatively high production costs due to low lipid productivity have been one of the major obstacles impeding their commercial production. We studied the effects of nitrogen sources and their concentrations on cell growth and lipid accumulation of Neochloris oleoabundans, one of the most promising oil-rich microalgal species. While the highest lipid cell content of 0.40 g/g was obtained at the lowest sodium nitrate concentration (3 mM), a remarkable lipid productivity of 0.133 g l⁻¹ day⁻¹ was achieved at 5 mM with a lipid cell content of 0.34 g/g and a biomass productivity of 0.40 g l⁻¹ day⁻¹. The highest biomass productivity was obtained at 10 mM sodium nitrate, with a biomass concentration of 3.2 g/l and a biomass productivity of 0.63 g l⁻¹ day⁻¹. It was observed that cell growth continued after the exhaustion of external nitrogen pool, hypothetically supported by the consumption of intracellular nitrogen pools such as chlorophyll molecules. The relationship among nitrate depletion, cell growth, lipid cell content, and cell chlorophyll content are discussed.