Application of Green Technology to Extract Clean and Safe Bioactive Compounds from Tetradesmus obliquus Biomass Grown in Poultry Wastewater.

Microalgae are capable of assimilating nutrients from wastewater (WW), producing clean water and biomass rich in bioactive compounds that need to be recovered from inside the microalgal cell. This work investigated subcritical water (SW) extraction to collect high-value compounds from the microalga Tetradesmus obliquus after treating poultry WW. The treatment efficiency was evaluated in terms of total Kjeldahl nitrogen (TKN), phosphate, chemical oxygen demand (COD) and metals. T. obliquus was able to remove 77% TKN, 50% phosphate, 84% COD, and metals (48-89%) within legislation values. SW extraction was performed at 170 °C and 30 bar for 10 min. SW allowed the extraction of total phenols (1.073 mg GAE/mL extract) and total flavonoids (0.111 mg CAT/mL extract) with high antioxidant activity (IC50 value, 7.18 µg/mL). The microalga was shown to be a source of organic compounds of commercial value (e.g., squalene). Finally, the SW conditions allowed the removal of pathogens and metals in the extracts and residues to values in accordance with legislation, assuring their safety for feed or agriculture applications.

Rhodosporidium toruloides and Tetradesmus obliquus Populations Dynamics in Symbiotic Cultures, Developed in Brewery Wastewater, for Lipid Production.

In this work, primary brewery wastewater (PBWW) and secondary brewery wastewater (SBWW) separately, or mixed at the ratios of 1:1 (PBWW:SBWW) and 1:7 (PBWW:SBWW), with or without supplementation with sugarcane molasses (SCM), were used as culture media for lipid production by a mixed culture of the oleaginous yeast Rhodosporidium toruloides NCYC 921 and the microalgae Tetradesmus obliquus (ACOI 204/07). Flow cytometry was used to understand the dynamics of the two micro-organisms during the mixed cultures evolution, as well as to evaluate the physiological states of each micro-organism, in order to assess the impact of the different brewery effluent media composition on the microbial consortium performance. Both brewery wastewaters (primary and secondary) without supplementation did not allow R. toruloides heterotrophic growth. Nevertheless, all brewery wastewater media, with and without SCM supplementation, allowed the microalgae growth, although the yeast was the dominant population. The maximum total biomass concentration of 2.17 g L-1 was achieved in the PBWW mixed cultivation with 10 g L-1 of SCM. The maximum lipid content (14.86% (w/w DCW)) was obtained for the mixed culture developed on SBWW supplemented with 10 g L-1 of SCM. This work demonstrated the potential of using brewery wastewater supplemented with SCM as a low-cost culture medium to grow R. toruloides and T. obliquus in a mixed culture for brewery wastewater treatment with concomitant lipid production.

Impact of High-Pressure Homogenization on the Cell Integrity of Tetradesmus obliquus and Seed Germination.

Microalgae have almost unlimited applications due to their versatility and robustness to grow in different environmental conditions, their biodiversity and variety of valuable bioactive compounds. Wastewater can be used as a low-cost and readily available medium for microalgae, while the latter removes the pollutants to produce clean water. Nevertheless, since the most valuable metabolites are mainly located inside the microalga cell, their release implies rupturing the cell wall. In this study, Tetradesmus obliquus grown in 5% piggery effluent was disrupted using high-pressure homogenization (HPH). Effects of HPH pressure (100, 300, and 600 bar) and cycles (1, 2 and 3) were tested on the membrane integrity and evaluated using flow cytometry and microscopy. In addition, wheat seed germination trials were carried out using the biomass at different conditions. Increased HPH pressure or number of cycles led to more cell disruption (75% at 600 bar and 3 cycles). However, the highest increase in wheat germination and growth (40-45%) was observed at the lowest pressure (100 bar), where only 46% of the microalga cells were permeabilised, but not disrupted. Non-treated T. obliquus cultures also revealed an enhancing effect on root and shoot length (up to 40%). The filtrate of the initial culture also promoted shoot development compared to water (21%), reinforcing the full use of all the process fractions. Thus, piggery wastewater can be used to produce microalgae biomass, and mild HPH conditions can promote cell permeabilization to release sufficient amounts of bioactive compounds with the ability to enhance plant germination and growth, converting an economic and environmental concern into environmentally sustainable applications.

Supercritical CO2 Extract from Microalga Tetradesmus obliquus: The Effect of High-Pressure Pre-Treatment.

High-pressure pre-treatment followed by supercritical carbon dioxide (ScCO2) extraction (300 bar, 40 °C) was applied for the attainment of the lipophilic fraction of microalga Tetradesmus obliquus. The chemical profile of supercritical extracts of T. obliquus was analyzed by ultra-high-performance liquid chromatography-high-resolution mass spectrometry with electrospray ionization (UHPLC-ESI-HRMS). Moreover, the impact of ScCO2 on the microbiological and metal profile of the biomass was monitored. The application of the pre-treatment increased the extraction yield approximately three-fold compared to the control. In the obtained extracts (control and pre-treated extracts), the identified components belonged to triacylglyceroles, fatty acid derivatives, diacylglycerophosphocholines and diacylglycerophosphoserines, pigments, terpenes, and steroids. Triacylglycerols (65%) were the most dominant group of compounds in the control extract. The pre-treatment decreased the percentage of triacylglycerols to 2%, while the abundance of fatty acid derivatives was significantly increased (82%). In addition, the pre-treatment led to an increase in the percentages of carotenoids, terpenoids, and steroids. Furthermore, it was determined that ScCO2 extraction reduced the number of microorganisms in the biomass. Considering its microbiological and metal profiles, the biomass after ScCO2 can potentially be used as a safe and important source of organic compounds.

Aquaculture wastewater treatment through microalgal. Biomass potential applications on animal feed, agriculture, and energy.

The use of microalgae to remediate raw effluent from brown crab aquaculture was evaluated by performing batch mode growth tests using separately the microalgae Chlorella vulgaris (Cv), Scenedesmus obliquus (Sc), Isochrysis galbana (Ig), Nannocloropsis salina (Ns), and Spirulina major (Sp). Removal efficiencies in batch growth were 100% for total nitrogen and total phosphorus for all microalgae. Chemical oxygen demand (COD) remediations were all above 72%. Biomass productivity varied from 20.9 mg L-1 day-1 (N. salina) to 146.4 mg L-1 day-1 (C. vulgaris). The two best performing algae were C. vulgaris and S. obliquus and they were tested in semi-continuous growth, reaching productivities of 879.8 mg L-1 day-1 and 811.7 mg L-1 day-1, respectively. The bioremediation of the effluent was tested with a transfer system consisting of three independent containers and compared with the use of a single container. The single container had the same capacity and received weekly the same volume of effluent as the three containers together. The remediation capacity of the 3 containers was much higher than the single one. The supplementation with NaNO3 was tested to improve the nutrient removal microalgae' capacity, with positive results. The removal efficiencies were 100% for total nitrogen and total phosphorus and higher than 96% for COD. The obtained C. vulgaris and S. obliquus biomass were composed of 31 and 35% proteins, 6 and 8% lipids, 39 and 30% carbohydrates, respectively. The composition of these biomass suggest that it can be used as novel and sustainable ingredients in aquaculture feeds. The algal biomass of Cv and Sc were used as biostimulants in the germination of wheat and watercress, and very promising results were attained, with increases in the germination index for Cv and Sc of 175% and 48% in watercress and 84% and 98% in wheat, respectively. The biomasses of Cv and Sc were also subjected to a torrefaction process with 72.5 ± 1.7% char yields. The obtained biochars were tested as biostimulants for germination seeds (wheat and watercress) and as bio-adsorbent of dye solutions.

Biostimulant Potential of Scenedesmus obliquus Grown in Brewery Wastewater.

Microalgae are microorganisms with the capacity to contribute to the sustainable and healthy food production, in addition to wastewater treatment. The subject of this work was to determine the potential of Scenedesmus obliquus microalga grown in brewery wastewater to act as a plant biostimulant. The germination index of watercress seeds, as well as the auxin-like activity in mung bean and cucumber, and in the cytokinin-like activity in cucumber bioassays were used to evaluate the biostimulant potential. Several biomass processes were studied, such as centrifugation, ultrasonication and enzymatic hydrolysis, as well as the final concentration of microalgal extracts to determine their influence in the biostimulant activity of the Scenedesmus biomass. The results showed an increase of 40% on the germination index when using the biomass at 0.1 g/L, without any pre-treatment. For auxin-like activity, the best results (up to 60% with respect to control) were obtained at 0.5 g/L of biomass extract, after a combination of cell disruption, enzymatic hydrolysis and centrifugation. For cytokinin-like activity, the best results (up to 187.5% with respect to control) were achieved without cell disruption, after enzymatic hydrolysis and centrifugation at a biomass extract concentration of 2 g/L.

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.

Combining biotechnology with circular bioeconomy: From poultry, swine, cattle, brewery, dairy and urban wastewaters to biohydrogen.

The ability of microalgae to grow in nutrient-rich environments and to accumulate nutrients from wastewaters (WW) makes them attractive for the sustainable and low-cost treatment of WW. The valuable biomass produced can be further used for the generation of bioenergy, animal feed, fertilizers, and biopolymers, among others. In this study, Scenedesmus obliquus was able to remove nutrients from different wastewaters (poultry, swine and cattle breeding, brewery and dairy industries, and urban) with removal ranges of 95-100% for nitrogen, 63-99% for phosphorus and 48-70% for chemical oxygen demand. The biomass productivity using wastewaters was higher (except for poultry) than in synthetic medium (Bristol), the highest value being obtained in brewery wastewater (1025 mg/(L.day) of freeze-dried biomass). The produced biomass contained 31-53% of proteins, 12-36% of sugars and 8-23% of lipids, regardless of the type of wastewater. The potential of the produced Scenedesmus obliquus biomass for the generation of BioH2 through batch dark fermentation processes with Enterobacter aerogenes was evaluated. The obtained yields ranged, in mL H2/g Volatile Solids (VS), from 50.1 for biomass from anaerobically digested cattle WW to 390 for swine WW, whereas the yield with biomass cultivated in Bristol medium was 57.6 mL H2/gVS.

Evaluation of Marine Microalga Diacronema vlkianum Biomass Fatty Acid Assimilation in Wistar Rats.

Diacronema vlkianum is a marine microalgae for which supposed health promoting effects have been claimed based on its phytochemical composition. The potential use of its biomass as health ingredient, including detox-shakes, and the lack of bioavailability studies were the main concerns. In order to evaluate the microalgae-biomass assimilation and its health-benefits, single-dose (CD1-mice) studies were followed by 66-days repeated-dose study in Wistar rats with the highest tested single-dose of microalgae equivalent to 101 mg/kg eicosapentaenoic acid + docosahexaenoic acid (EPA+DHA). Microalgae-supplementation modulated EPA and docosapentaenoic acid enrichment at arachidonic acid content expenditure in erythrocytes and liver, while increasing EPA content of heart and adipose tissues of rats. Those fatty acid (FA) changes confirmed the D. vlkianum-biomass FA assimilation. The principal component analyses discriminated brain from other tissues, which formed two other groups (erythrocytes, liver, and heart separated from kidney and adipose tissues), pointing to a distinct signature of FA deposition for the brain and for the other organs. The improved serum lipid profile, omega-3 index and erythrocyte plasticity support the cardiovascular benefits of D. vlkianum. These results bolster the potential of D. vlkianum-biomass to become a "heart-healthy" food supplement providing a safe and renewable source of bioavailable omega-3 FA.

Integrated microbial processes for biofuels and high value-added products: the way to improve the cost effectiveness of biofuel production.

The production of microbial biofuels is currently under investigation, as they are alternative sources to fossil fuels, which are diminishing and their use has a negative impact on the environment. However, so far, biofuels derived from microbes are not economically competitive. One way to overcome this bottleneck is the use of microorganisms to transform substrates into biofuels and high value-added products, and simultaneously taking advantage of the various microbial biomass components to produce other products of interest, as an integrated process. In this way, it is possible to maximize the economic value of the whole process, with the desired reduction of the waste streams produced. It is expected that this integrated system makes the biofuel production economically sustainable and competitive in the near future. This review describes the investigation on integrated microbial processes (based on bacteria, yeast, and microalgal cultivations) that have been experimentally developed, highlighting the importance of this approach as a way to optimize microbial biofuel production process.

Resources recovery from domestic wastewater by a combined process: anaerobic digestion and membrane photobioreactor.

Anaerobic and membrane technologies are a promising combination to decrease the energy consumption associated with wastewater treatment, allowing the recovery of resources: organic matter as biomethane, nutrient assimilation by microalgae and reclaimed water. In this study, domestic wastewater was treated using a combination of an upflow anaerobic sludge blanket sludge reactor (UASB) and a membrane photobioreactor (MPBR). The outdoor facilities were operated continuously for three months under unfavourable environmental conditions such as lack of temperature control, winter season with lower solar irradiation and lower daylight hours which was a challenge for the present work, not previously described. The energetic valorisation of the organic matter present in the wastewater by biomethane produced in the UASB would contribute to reducing overall facilities' energy requirements. The ultrafiltration (UF) membrane facilitated the harvesting of biomass, operating at 10 L·h-1·m-2 during the experimental period. Although the main contribution to fouling was irreversible, chemical cleanings were not necessary due to effective fouling control, which prevented the final TMP from exceeding 25 kPa. In addition, microalgae-bacterial consortium developed without prior inoculation were harvested from the MPBR using membrane assistance. The obtained biomass was also successfully tested as a biostimulant for corn germination/growth, as well as a biopesticide against Rhizoctonia solani and Fusarium oxysporum.

Supercritical extraction of lycopene from tomato industrial wastes with ethane.

Supercritical fluid extraction of all-E-lycopene from tomato industrial wastes (mixture of skins and seeds) was carried out in a semi-continuous flow apparatus using ethane as supercritical solvent. The effect of pressure, temperature, feed particle size, solvent superficial velocity and matrix initial composition was evaluated. Moreover, the yield of the extraction was compared with that obtained with other supercritical solvents (supercritical CO2 and a near critical mixture of ethane and propane). The recovery of all-E-lycopene increased with pressure, decreased with the increase of the particle size in the initial stages of the extraction and was not practically affected by the solvent superficial velocity. The effect of the temperature was more complex. When the temperature increased from 40 to 60 °C the recovery of all-E-lycopene increased from 80 to 90%. However, for a further increase to 80 °C, the recovery remained almost the same, indicating that some E-Z isomerization could have occurred, as well as some degradation of lycopene. The recovery of all-E-lycopene was almost the same for feed samples with different all-E-lycopene content. Furthermore, when a batch with a higher all-E-lycopene content was used, supercritical ethane and a near critical mixture of ethane and propane showed to be better solvents than supercritical CO2 leading to a faster extraction with a higher recovery of the carotenoid.

Production of Mannosylerythritol Lipids Using Oils from Oleaginous Microalgae: Two Sequential Microorganism Culture Approach.

Mannosylerythritol lipids (MELs) are biosurfactants with excellent biochemical properties and a wide range of potential applications. However, most of the studies focusing on MELs high titre production have been relying in the use of vegetable oils with impact on the sustainability and process economy. Herein, we report for the first time MELs production using oils produced from microalgae. The bio-oil was extracted from Neochloris oleoabundans and evaluated for their use as sole carbon source or in a co-substrate strategy, using as an additional carbon source D-glucose, on Moesziomyces spp. cultures to support cell growth and induce the production of MELs. Both Moesziomyces antarcticus and M. aphidis were able to grow and produce MELs using algae-derived bio-oils as a carbon source. Using a medium containing as carbon sources 40 g/L of D-glucose and 20 g/L of bio-oils, Moesziomyces antarcticus and M. aphidis produced 12.47 ± 0.28 and 5.72 ± 2.32 g/L of MELs, respectively. Interestingly, there are no significant differences in productivity when using oils from microalgae or vegetable oils as carbon sources. The MELs productivities achieved were 1.78 ± 0.04 and 1.99 ± 0.12 g/L/h, respectively, for M. antarcticus fed with algae-derived or vegetable oils. These results open new perspectives for the production of MELs in systems combining different microorganisms.

Nannochloropsis oceanica harvested using electrocoagulation with alternative electrodes - An innovative approach on potential biomass applications.

Electrocoagulation is a promising technology to harvest microalgal biomass. However, the commonly used aluminum electrodes release undesired salts that decrease biomass value. In this study, alternative iron, zinc, and magnesium electrodes and operational parameters pH, time and current density were studied to harvest Nannochloropsis oceanica. For recovery efficiency and concentration factor the initial pH was most important using iron electrodes, while time and current density were more relevant using zinc and magnesium electrodes. Optimal parameters resulted in biomass recovery efficiencies > 95%, biomass was concentrated 2.8-7.2 times and contained 15.7-29.1% ashes. Elemental analysis revealed metal salts in harvested biomass resulting from electrode corrosion. Finally, ash contents could be reduced by 65% using EDTA as a chelating agent. The electrocoagulation harvested microalgal biomass enriched in essential metals may be a promising bioresource for agricultural growth inducers, or functional ingredients for feed.

Algae as Food in Europe: An Overview of Species Diversity and Their Application.

Algae have been consumed for millennia in several parts of the world as food, food supplements, and additives, due to their unique organoleptic properties and nutritional and health benefits. Algae are sustainable sources of proteins, minerals, and fiber, with well-balanced essential amino acids, pigments, and fatty acids, among other relevant metabolites for human nutrition. This review covers the historical consumption of algae in Europe, developments in the current European market, challenges when introducing new species to the market, bottlenecks in production technology, consumer acceptance, and legislation. The current algae species that are consumed and commercialized in Europe were investigated, according to their status under the European Union (EU) Novel Food legislation, along with the market perspectives in terms of the current research and development initiatives, while evaluating the interest and potential in the European market. The regular consumption of more than 150 algae species was identified, of which only 20% are approved under the EU Novel Food legislation, which demonstrates that the current legislation is not broad enough and requires an urgent update. Finally, the potential of the European algae market growth was indicated by the analysis of the trends in research, technological advances, and market initiatives to promote algae commercialization and consumption.

Incorporation of Chlorella vulgaris and Spirulina maxima biomass in pasta products. Part 1: Preparation and evaluation.

BACKGROUND: Microalgae are able to enhance the nutritional content of conventional foods and hence to positively affect human health, due to their original chemical composition. The aim of the present study was to prepare fresh spaghetti enriched with different amounts of microalgae biomass (Chlorella vulgaris and Spirulina maxima) and to compare the quality parameters (optimal cooking time, cooking losses, swelling index and water absorption), chemical composition, instrumental texture and colour of the raw and cooked pasta enriched with microalgae biomass with standard semolina spaghetti. RESULTS: The incorporation of microalgae results in an increase of quality parameters when compared to the control sample. The colour of microalgae pastas remained relatively stable after cooking. The addition of microalgae resulted in an increase in the raw pasta firmness when compared to the control sample. Of all the microalgae studied, an increase in the biomass concentration (0.5-2.0%) resulted in a general tendency of an increase in the pasta firmness. Sensory analysis revealed that microalgae pastas had higher acceptance scores by the panellists than the control pasta. CONCLUSION: Microalgae pastas presented very appellative colours, such as orange and green, similar to pastas produced with vegetables, with nutritional advantages, showing energetic values similar to commercial pastas. The use of microalgae biomass can enhance the nutritional and sensorial quality of pasta, without affecting its cooking and textural properties.

Microalgae as a raw material for biofuels production.

Biofuels demand is unquestionable in order to reduce gaseous emissions (fossil CO(2), nitrogen and sulfur oxides) and their purported greenhouse, climatic changes and global warming effects, to face the frequent oil supply crises, as a way to help non-fossil fuel producer countries to reduce energy dependence, contributing to security of supply, promoting environmental sustainability and meeting the EU target of at least of 10% biofuels in the transport sector by 2020. Biodiesel is usually produced from oleaginous crops, such as rapeseed, soybean, sunflower and palm. However, the use of microalgae can be a suitable alternative feedstock for next generation biofuels because certain species contain high amounts of oil, which could be extracted, processed and refined into transportation fuels, using currently available technology; they have fast growth rate, permit the use of non-arable land and non-potable water, use far less water and do not displace food crops cultures; their production is not seasonal and they can be harvested daily. The screening of microalgae (Chlorella vulgaris, Spirulina maxima, Nannochloropsis sp., Neochloris oleabundans, Scenedesmus obliquus and Dunaliella tertiolecta) was done in order to choose the best one(s), in terms of quantity and quality as oil source for biofuel production. Neochloris oleabundans (fresh water microalga) and Nannochloropsis sp. (marine microalga) proved to be suitable as raw materials for biofuel production, due to their high oil content (29.0 and 28.7%, respectively). Both microalgae, when grown under nitrogen shortage, show a great increase (approximately 50%) in oil quantity. If the purpose is to produce biodiesel only from one species, Scenedesmus obliquus presents the most adequate fatty acid profile, namely in terms of linolenic and other polyunsaturated fatty acids. However, the microalgae Neochloris oleabundans, Nannochloropsis sp. and Dunaliella tertiolecta can also be used if associated with other microalgal oils and/or vegetable oils.

Solar Pond devices: free energy or bioreactors for Artemia biomass production?

The recent exponential growth in industrial aquaculture has led to a huge increase in Artemia biomass production in order to meet increased fish production needs. The present study explores the potential use of salt gradient solar ponds (SGSPs) for production of Artemia nauplii. An SGSP is a basin of water where solar energy is trapped and collected via an artificially imposed gradient. Three zones can be identified in an SGSP: upper and lower zones, which are both convective, and a middle zone, which is intended to be non-convective. The latter acts as a transparent insulation layer and allows for storage of solar energy at the bottom, where it is available for use. The combination of salt, temperature and high transparency could make SGSPs promising bioreactors for the production of Artemia nauplii. Using particle image velocymetry (PIV) and Shadowgraph visualisation techniques, the behaviour of Artemia nauplii under critical cultivation parameters (namely, salinity, temperature and light) was monitored to determine movement velocity, and how movement of Artemia affects the salt gradient. It was observed that Artemia nauplii constantly follow light, irrespective of adverse salinity and/or temperature conditions. However, despite the substantial displacement of Artemia following the light source, the salt gradient is not disrupted. The suitability of SGSPs as bioreactors for Artemia biomass production was then tested. The results were disappointing, probably due to the lack of sufficient O(2) for Artemia survival and growth. Follow-up trials were conducted aimed at using the SGSP as a green and economically attractive energy source to induce faster hatching of cysts and improved Artemia nauplii growth. The results of these trials, and a case study of Artemia nauplii production using an SGSP, are presented. The authors constructed a Solar Pond device, which they suggest as a novel way of supplying thermal energy for Artemia biomass production in an aquaculture enterprise. Finally, the authors suggest a method of producing and collecting Artemia biomass, and of heating a fish larval tank, in an 'ideal' Solar Pond device, profiting from the low investment costs of using a decommissioned salt works.

Neochloris oleabundans UTEX #1185: a suitable renewable lipid source for biofuel production.

Energy crises, global warming, and climatic changes call for technological and commercial advances in manufacturing high-quality transportation fuels from unconventional feedstocks. Microalgae is one of the most promising sources of biofuels due to the high yields attained per unit area and because it does not displace food crops. Neochloris oleabundans (Neo) microalga is an important promising microbial source of single-cell oil (SCO). Different experimental growth and lipid production conditions were evaluated and compared by using optical density (540 nm), dry-weight determination, and flow cytometry (FC). Best Neo average biomass productivity was obtained at 30 degrees C under conditions of nitrogen-sufficiency and CO(2) supplementation (N+/30 degrees C/CO(2)), with an average doubling time of 1.4 days. The second and third highest productivities occurred with N-sufficient cultures without CO(2) supplementation at 26 degrees C (N+/26 degrees C) and at 30 degrees C (N+/30 degrees C), with doubling times of 1.7 and 2.2 days, respectively. Microbial lipid production was monitored by flow cytometry using Nile red (NR), a lipophilic fluorochrome that possesses several advantageous characteristics for in situ screening near real time (at line). Results showed maximum lipid content (56%) after 6 days of nitrogen depletion under nitrogen starvation without CO(2) supplementation (N-/30 degrees C), followed by N-/30 degrees C/CO(2) and N-/26 degrees C conditions with 52% lipid content, after 5 and 6 days of N starvation, respectively. The adequate fatty acid profile and iodine value of Neo lipids reinforced this microalga as a good source of SCO, in particular for use as biodiesel.

Scenedesmus obliquus in poultry wastewater bioremediation.

Wastewater biological treatment with microalgae can be an effective technology, removing nutrients and other contaminants while reducing chemical oxygen demand. This can be particularly interesting for the meat producing industry which produces large volumes of wastewater from the slaughtering of animals and cleaning of their facilities. The main purpose of this research was the treatment of poultry wastewater using Scenedesmus obliquus in an economical and environmentally sustainable way. Two wastewaters were collected from a Portuguese poultry slaughterhouse (poultry raw - PR and poultry flocculated - PF) and the bioremediation was evaluated. The performance of microalga biomass growth and biochemical composition were assessed for two illumination sources (fluorescent vs LEDs). S. obliquus achieved positive results when grown in highly contaminated agro-industrial wastewater from the poultry industry, independently of the light source. The wastewater bioremediation revealed results higher than 97% for both ammonium and phosphate removal efficiency, for a cultivation time of 13 days. The saponifiable matter obtained from the biomass of the microalga cultures was, on average, 11% and 27% (m/malga) with PR and PF wastewater, respectively. In opposition, higher sugar content was obtained from microalgae biomass grown in PR wastewater (average 34% m/malga) in comparison to PF wastewater (average 23% m/malga), independently of the illumination source. Therefore, biomass obtained with PR wastewater will be more appropriate as a raw material for bioethanol/biohydrogen production (higher sugar content) while biomass produced in PF wastewater will have a similar potential as feedstock for both biodiesel and bioethanol/biohydrogen production (similar lipid and sugar content).