Cell wall disruption: An effective strategy to improve the nutritive quality of microalgae in African catfish (Clarias gariepinus)
Agboola, Jeleel O. ; Teuling, Emma ; Wierenga, Peter A. ; Gruppen, Harry ; Schrama, Johan W. - \ 2019
Aquaculture Nutrition 25 (2019)4. - ISSN 1353-5773 - p. 783 - 797.
accessibility - algae - digestibility - disruption treatments - nutrient utilization - rigid cell wall
The rigid cell walls of microalgae may hinder their utilization in fish feeds. The current experiment assessed the correlation between the accessibility of microalgae nutrients and their in vivo digestibility in African catfish. Nannochloropsis gaditana biomass was subjected to physical or mechanical treatments to weaken its cell wall; untreated—no disruption treatment (UNT), pasteurization (PAS), freezing (FRO), freeze-drying (FRD), cold pasteurization (L40) and bead milling (BEM). Six experimental diets formulated from differently treated and untreated microalgae (at 30% diet inclusion level) were tested on growth performance and apparent nutrient digestibility (ADCs) in juvenile African catfish. A basal diet (REF) containing no microalgae was used as reference diet. Results showed that biomass gain and feed conversion ratio of fish fed L40 and BEM diets increased by 13% and 11%, respectively, relative to the UNT diet. Additionally, FRD, FRO, L40 and BEM cell wall disruption treatments improved protein digestibility by 0.5%, 5.9%, 8.4% and 16.3%, respectively, compared to the UNT treatment. There was a positive correlation between accessibility of microalgal nutrients and their digestibility in African catfish. Nutrient digestibility of microalgae was dependent on extent of cell disruption. Also, the impact of cell disruption on nutrient digestibility of microalgae differs between African catfish and Nile tilapia.
Stof tot nadenken: vier manieren om de mode-industrie duurzamer te maken
Louwerens, Tessa ; Poldner, Kim ; Fischer, A.R.H. ; Broek, L.A.M. van den; Dam, J.E.G. van; Houthoff, Iris - \ 2018
biobased materials - clothing - fibres - algae - biomass - hemp - dyes - pigments - bioplastics - biopolymers - ink - natural products
Instead of eating herring we should eat algae!
Janssen, M.G.J. - \ 2017
Wageningen : WURcast
What makes algae a sufficient food?
Final report: Environmental assessment of algae-based PUFA production
Keller, H. ; Reinhardt, G. ; Rettenmaier, N. ; Schorb, A. ; Dittrich, M. ; Wolf, P.L. de; Voort, M.P.J. van der; Spruijt, J. ; Potters, J.I. ; Elissen, H.J.H. - \ 2017
Heidelberg : PUFAChain - 94
algae - biofuels - bioenergy - biobased economy - biomass - omega-3 fatty acids - plant oils - biobased chemistry - fermentation
Socio-economic assessment of Algae-based PUFA production : The value chain from microalgae to PUFA ('PUFACHAIN')
Voort, M.P.J. van der; Spruijt, J. ; Potters, J.I. ; Elissen, H.J.H. - \ 2017
Göttingen : PUFAChain - 84
biobased economy - biomass - bioenergy - biofuels - algae - plant oils - omega-3 fatty acids - economic analysis
Socio-economic assessment of Algae-based PUFA production
Voort, Marcel van der; Spruijt, Joanneke ; Potters, Jorieke ; Wolf, Pieter de; Elissen, Hellen - \ 2017
Göttingen : PUFAChain - 84
bioenergy - biobased economy - biofuels - biomass - algae - fatty acids - bio-energie - biobased economy - biobrandstoffen - biomassa - algen - vetzuren
Botryococcus braunii for the production of hydrocarbons and exopolysaccharides and the role of associated bacteria
Gouveai, João Diogo Guimarães - \ 2017
Wageningen University. Promotor(en): R.H. Wijffels, co-promotor(en): M.J. Barbosa; D. Sipkema. - Wageningen : Wageningen University - ISBN 9789463436960 - 157
biomass production - algae - algae culture - hydrocarbons - bacteria - biomassa productie - algen - algenteelt - koolwaterstoffen - bacteriën
Microalgae are photosynthetic organisms that are found worldwide in many different aquatic environments and therefore display an immense biological diversity. They are a promising source of many useful polymers that have industrial applications such as food, fuel, material and pharmaceutical. One microalga that has gathered quite a research community is Botryococcus braunii. The reason for its scientific club is the fact it can synthetize long chain hydrocarbons molecules from C20 to C40. These hydrocarbons have been found in oil-shales and tests show that it can be used as aviation fuel. Besides producing hydrocarbons, some strains of B. braunii can produce exopolysaccharides (EPS) composed mainly of galactose and a small fraction of fucose. The EPS has interesting rheological properties for the food industry and potential active compounds that could be used in the pharmaceutical industry .
Like many other microorganisms, microalgae in the natural environment are usually in the presence of bacteria. The presence of bacteria with microalgae can either have a beneficial or an antagonistic effect. For B. braunii little is known about the bacteria community present especially for the EPS producing strain. For that reason, the aim of this thesis was to investigate B. braunii’s associated bacteria with the aim of improving B. braunii’s biomass growth and hydrocarbon and EPS content. In chapter one, we introduced the topic of microalgae as a potential source of sustainable polymers and we introduced the species B. braunii, describing its characteristics and scientific interest. It is also introduced the topic of microalgae and bacteria associations by looking at other studies from literature.
In chapter two, 16 publically available strains of B. braunii were ordered in culture banks and screened for biomass productivity, hydrocarbon and total carbohydrate content. The aim of the study was to identify one or more good strains that displayed high biomass productivity as well as hydrocarbon or total carbohydrate content. In seven strains out of 16 cultivated in 250 mL volume Erlenmeyer flaks, we detected 5 to 42 % content of hydrocarbons of the dry biomass with four strains producing botryococcenes (C30-C34) and three strains producing alkanes (C20-C25). Two strains showed high amounts of EPS content above 50 % per dry biomass. Seven strains comprising of the strains with higher biomass productivity plus the highest hydrocarbons and EPS content, were tested for scalability using bench scale 800 mL volume bubble column reactors. Two strains, AC761 which produces botryococcenes and CCALA778 which produces EPS, were selected as the most promising B. braunii strains for industrial production of hydrocarbon and EPS.
In chapter three, we studied the bacterial community associated with B. braunii. We cultured 12 strains from the initial 16 from chapter 2 and extracted the DNA from samples taken over a time period of 12 days. It was clear from this study that B. braunii hosts a variety of bacterial species and still maintain its growth. The bacteria families Rhizobiaceae, Bradyrhizobiaceae and Comamonadaceae were found in all 12 strains. These families which belong to the phylum Proteobacteria could have an important role regarding B. braunii growth. Each strain displayed a different bacterial community composition but all the strains from the CAEN culture collection clustered near each other suggesting that the algae culture collection could have an influence on the bacterial community composition. Bacteria genus identification based on 16S rRNA gene amplicon similarity showed several genera present including Rhizobium spp. and Variovorax spp.. Two genera were found that are possibly linked to hydrocarbon degradation: Sphingomonas spp. and Rhodobacter spp..
In chapter four, we investigated further B. braunii CCALA778 which was shown in chapter 2 to accumulate high amounts of EPS. We investigated the effects of antibiotics on algal growth, EPS accumulation and bacterial community composition of CCALA778. Taxonomical identification by 16S rRNA gene analysis indicated that most of the bacteria present with CCALA778 were Gram-negative. Of all antibiotics and antibiotic mixes, only the treatment with Penicillin did not affect the growth of B. braunii. The remaining antibiotics halted the growth of CCALA778 while they were active. The exceptions were with the antibiotics Chloramphenicol, Gentamycin and Linezolid which permanently ceased the growth of CCALA778. The accumulation of EPS seemed to be related to biomass growth, but we did also observe a reduction of EPS with the cultures treated with Penicillin suggesting that bacteria could have an effect on the EPS content. Antibiotics had specific effects on the bacterial community with all treatments showing significant changes over time. The most efficient treatment in removing bacteria were the mixes Metronidazole-Rifampicin-Penicillin and Penicillin-Rifampicin which were the only treatments to show significant changes in the bacterial community when compared to the untreated cultures after 10 days of cultivation. Antibiotics and antibiotic mixes can create changes in the bacterial community but it is unlikely that they alone can lead to axenic B. braunii cultures.
In chapter five, we used Ultra Violet-C light (UVC) to reduce bacteria diversity and abundance present in B. braunii CCALA778. UVC is highly effective in inactivating bacteria and for that reason is being investigated further in medicinal applications. After applying the UVC to B. braunii CCALA778, we were able to reduce the relative abundance of 16S rRNA genes assigned to bacteria to less than 1 % compared to the 70% in the non-treated cultures. With the UVC treated CCALA778 we observed several physiological changes. The UV treated cultures with reduced bacterial load showed nearly double the EPS accumulation when compared to the untreated. To confirm that we did not see an artefact in our results due to the UVC treatment, UVC treated cultures were also inoculated with bacteria from the untreated and we observed a reduction of EPS similar to what we saw with the untreated cultures. There were no changes to the EPS composition after the removal of the bacteria. Other physiological changes were observed, namely that colony size of B. braunii CCALA778 significantly increased when compared to the untreated culture and the UV treated with bacteria. We hypothesise that the increase in colony size was probably due to the fact there was more EPS accumulated which helped with cell aggregation. We also observed an increase on the biomass growth in the UV-treated CCALA778 which we hypothesized being related to the fact that there was none or hardly any competition for essential micronutrients such as phosphate. From this study we concluded that the associated bacteria present with B. braunii CCALA778 were antagonistic. We believe the reason why the bacteria were antagonistic is because of the readily available EPS which is a rich source of organic compounds that bacteria could use for their own proliferation allowing them to compete with B. braunii for essential nutrients.
In chapter 6, we discuss the implications from our previous 4 experimental chapters. The aim of the study was to improve the biomass productivity and hydrocarbon and EPS content of the microalgae B. braunii. In brief, B. braunii displayed a wide range of physiological traits regarding biomass productivity and hydrocarbon and total carbohydrate content. We showed that B. braunii can co-habit with a wide range of bacteria diversity and abundance and that the associated bacteria were antagonistic to CCALA778 by affecting its biomass growth. We also showed that by removing the associated bacteria we can increase the EPS accumulation. Currently most of the research on microalgae and bacteria interactions, focus on the positive side, but we must understand also how bacteria can be antagonistic to microalga growth. Bacteria can be antagonistic to microalgae by competing for nutrients and also being detrimental to industrial process by degrading the product of interest in the case of organic carbons such as EPS. Therefore it is unlikely we can use the benefits that bacteria can provide such as enhancing growth to improve the cultivation of B. braunii and other similar microalgae species that secrete EPS. Since bacteria can be antagonistic to microalgae that secrete large amounts of organic compounds such as EPS, it is imperative to minimize contamination in large scale photobioreactors (PBR). It is important because in large scale PBR, contamination can occur leading to downtime of the reactors. If microalgae industry is to advance, it must develop PBR units that prevent contamination of bacteria from the surrounding environment.
Measurements of transcripts, proteome and metabolite profiles
Peters, Sander - \ 2017
Fuel4Me - 7
biobased economy - biobrandstoffen - algen - algenteelt - oliën - biobased economy - biofuels - algae - algae culture - oils
The work described for this deliverable was carried out by DLO-PRI, in collaboration with WU, and aim ed to understanding lipid production in Phaeodactylum tricornutum . By means of transcriptome, proteome and metabolome analyses we aimed to provide insight into the one step lipid production performed by partner WU. Cultivation conditions were chosen to steer lipid profile towards most suitable composition for biofuel production.
Photosynthetic efficiency in microalgal lipid production
Remmers, Ilse M. - \ 2017
Wageningen University. Promotor(en): R.H. Wijffels, co-promotor(en): P.P. Lamers. - Wageningen : Wageningen University - ISBN 9789463434607 - 200
algae - biofuels - light - triacylglycerols - lipids - metabolism - algae culture - cultural methods - algen - biobrandstoffen - licht - triacylglycerolen - lipiden - metabolisme - algenteelt - cultuurmethoden
Microalgae can contain large amounts of lipids which make them a promising feedstock for sustainable production of food, feed, fuels and chemicals. Various studies, including pilot-scale, have been performed and the knowledge on microalgal processes has advanced quickly. Unfortunately, current production costs for cultivation are still too high for bulk lipid production from microalgae.
One of the major causes for the high costs of bulk lipid production is the reduced solar-to-lipid conversion efficiency. Current research, however, does not provide sufficient insight to identify optimization targets. Therefore, in this thesis we have studied the lipid production in microalgae in depth.
Different TAG-accumulation strategies were investigated from a process engineering and metabolic point of view. The combination of all findings were used in the general discussion to thoroughly evaluate the microalgal lipid accumulation strategies. Current phototrophic microalgal lipid yields are still 10 times lower than the theoretical maximum. There is, however, still an enormous potential for further improvements. Future research should focus on (genetically) improved strains and advanced cultivation strategies, including adaptation to fluctuating outdoor weather conditions.
This thesis was performed within the EU FP7 FUEL4ME project under grand agreement No 308938. Objective of this program is to develop a sustainable and scalable process for biofuels from microalgae and to valorize the by-products.
Metabolic modeling to understand and redesign microbial systems
Heck, Ruben G.A. van - \ 2017
Wageningen University. Promotor(en): V.A.P. Martins dos Santos, co-promotor(en): M. Suárez Diez. - Wageningen : Wageningen University - ISBN 9789463434553 - 239
micro-organismen - modelleren - kooldioxide - biotechnologie - algen - metabolisme - pseudomonas - microorganisms - modeling - carbon dioxide - biotechnology - algae - metabolism - pseudomonas
The goals of this thesis are to increase the understanding of microbial metabolism and to functionally (re-)design microbial systems using Genome- Scale Metabolic models (GSMs). GSMs are species-specific knowledge repositories that can be used to predict metabolic activities for wildtype and genetically modified organisms. Chapter 1 describes the assumptions associated with GSMs, the GSM generation process, common GSM analysis methods, and GSM-driven strain design methods. Thereby, chapter 1 provides a background for all other chapters. In this work, there is a focus on the metabolically versatile bacterium Pseudomonas putida (chapters 2,3,4,5,6), but also other model microbes and biotechnologically or societally relevant microbes are considered (chapters 3,4,6,7,8).
GSMs are reflections of the genome annotation of the corresponding organism. For P. putida, the genome annotation that GSMs have been built on is more than ten years old. In chapter 2, this genome annotation was updated both on a structural and functional level using state-of-the-art annotation tools. A crucial part of the functional annotation relied on the most comprehensive P. putida GSM to date. This GSM was used to identify knowledge gaps in P. putida metabolism by determining the inconsistencies between its growth predictions and experimental measurements. Inconsistencies were found for 120 compounds that could be degraded by P. putida in vitro but not in silico. These compounds formed the basis for a targeted manual annotation process. Ultimately, suitable degradation pathways were identified for 86/120 as part of the functional reannotation of the P. putida genome.
For P. putida there are 3 independently generated GSMs, which is not uncommon for model organisms. These GSMs differ in generation procedure and represent different and complementary subsets of the knowledge on the metabolism of the organism. However, the differing generation procedures also makes it extremely cumbersome to compare their contents, let alone to combine them into a single consensus GSM. Chapter 3 addresses this issue through the introduction of a computational tool for COnsensus Metabolic Model GENeration (COMMGEN). COMMGEN automatically identifies inconsistencies between independently generated GSMs and semi-automatically resolves them. Thereby, it greatly facilitates a detailed comparison of independently generated GSMs as well as the construction of consensus GSMs that more comprehensively describe the knowledge on the modeled organism.
GSMs can predict whether or not the corresponding organism and derived mutants can grow in a large variety of different growth conditions. In comparison, experimental data is extremely limited. For example, BIOLOG data describes growth phenotypes for one strain in a few hundred different media, and genome-wide gene essentially data is typically limited to a single growth medium. In chapter 4 GSMs of multiple Pseudomonas species were used to predict growth phenotypes for all possible single-gene-deletion mutants in all possible minimal growth media to determine conditionally and unconditionally essential genes. This simulated data was integrated with genomic data on 432 sequenced Pseudomonas species, which revealed a clear link between the essentiality of a gene function and the persistence of the gene within the Pseudomonas genus.
Chapters 5 and 6 describe the use of GSMs to (re-)design microbial systems. P. putida is, despite its acknowledged versatile metabolism, an obligate aerobe. As the oxygen-requirement limits the potential applications of P. putida, there have been several experimental attempts to enable it to grow anaerobically, which have so far not succeeded. Chapter 5 describes an in silico effort to determine why P. putida cannot grow anaerobically using a combination of GSM analyses and comparative genomics. These analyses resulted in a shortlist of several essential and oxygen-dependent processes in P. putida. The identification of these processes has enabled the design of P. putida strains that can grow anaerobically based on the current understanding of P. putida metabolism as represented in GSMs.
Efficient microbial CO2 fixation is a requirement for the biobased community, but the natural CO2 fixation pathways are rather inefficient, while the synthetic CO2 fixation pathways have been designed without considering the metabolic context of a target organism. Chapter 6 introduces a computational tool, CO2FIX, that designs species-specific CO2 fixation pathways based on GSMs and biochemical reaction databases. The designed pathways are evaluated for their ATP efficiency, thermodynamic feasibility, and kinetic rates. CO2FIX is applied to eight different organisms, which has led to the identification of both species-specific and general CO2 fixation pathways that have promising features while requiring surprisingly few non-native reactions. Three of these pathways are described in detail.
In all previous chapters GSMs of relatively well-understood microbes have been used to gain further insight into their metabolism and to functionally (re-)design them. For complex microbial systems, such as algae (chapter 7) and gut microbial communities (chapter 8), GSMs are similarly useful, but substantially more difficult to create and analyze. Algae are widely considered as potential centerpieces of a biobased economy. Chapter 7 reviews the current challenges in algal genome annotation, modeling and synthetic biology. The gut microbiota is an incredibly complex microbial system that is crucial to our well-being. Chapter 8 reviews the ongoing developments in the modeling of both single gut microbes and gut microbial communities, and discusses how these developments will enable the move from studying correlation to causation, and ultimately the rational steering of gut microbial activity.
Chapter 9 discusses how the previous chapters contribute to the research goals of this thesis. In addition, it provides an extensive discussion on current GSM practices, the issues associated therewith, and how these issues can be tackled. In particular, the discussion focuses on issues related to: (i) The inability to distinguish between biological difference and GSM generation artifacts when using multiple GSMs, (ii) The lack of continuous GSM updates, (iii) The mismatch between what GSM predictions and experimental data represent, (iv) The need for standardization in GSM evaluation, and (v) The lack of experimental validation of GSM-driven strain design for metabolic engineering.
A blooming business : Identifying limits to Lake Taihu's nutrient input
Janssen, Annette B.G. - \ 2017
Wageningen University. Promotor(en): Wolf Mooij, co-promotor(en): J.H. Janse; A.A. van Dam. - Wageningen : Wageningen University - ISBN 9789463431897 - 268
lakes - freshwater ecology - aquatic ecosystems - nutrients - cycling - nutrient flows - biodiversity - algae - models - critical loads - limnology - spatial variation - ecological restoration - china - meren - zoetwaterecologie - aquatische ecosystemen - voedingsstoffen - kringlopen - nutriëntenstromen - biodiversiteit - algen - modellen - critical loads - limnologie - ruimtelijke variatie - ecologisch herstel - china
Last century, Lake Taihu (China) became serious eutrophic due to excessive nutrient input. During the 1980s, the first algal blooms emerged in the lake, reaching disastrous proportions in 2007. During that year, the intake of drinking water had to be shut down and millions of people had to look for an alternative source of drinking water. This raises the question whether such problems can be avoided. Of crucial importance in avoiding and reducing toxic algal blooms is the identification of the maximum nutrient load ecosystems can absorb, while remaining in a good ecological state. In this thesis, I aim to determine the critical nutrient load for Lake Taihu. I approach the search for critical nutrient loads of Lake Taihu in five steps with diversity as an overarching topic throughout this thesis: diversity in lakes, diversity in models, diversity in spatial distribution of nutrient and water sources, diversity in the development of lakes around the earth and finally diversity within specific lakes. From the long list of available models I chose the model PCLake to use in my analysis because it is the most extensively used food web model applied for bifurcation analysis of shallow aquatic ecosystems. The approach has resulted in a range of critical nutrient loads for different parts of Lake Taihu. Furthermore, critical nutrient loads depend on management goals, i.e. the maximum allowable chlorophyll-a concentration. According to the model results, total nutrient loads need to be more than halved to reach chlorophyll-a concentrations of 30-40 μg.L-1 in most sections of the lake. To prevent phytoplankton blooms with 20 μg.L-1 chlorophyll-a throughout Lake Taihu, both phosphorus and nitrogen loads need a nearly 90% reduction. This range contrasts to the single point of recovery that is often found for small shallow lakes. The range in critical nutrient loads found for Lake Taihu can be interpreted as providing a path of recovery for which each step leads to water quality improvement in certain parts of the lake. To reach total recovery, nutrient reduction seems to be the most promising management option.
Harvesting and cell disruption of microalgae
Lam, Gerard Pieter 't - \ 2017
Wageningen University. Promotor(en): R.H. Wijffels; M.H.M. Eppink, co-promotor(en): M.H. Vermuë. - Wageningen : Wageningen University - ISBN 9789463431736 - 206
algae - harvesting - flocculation - polymers - chlorella vulgaris - biorefinery - electric field - organelles - algen - oogsten - uitvlokking - polymeren - chlorella vulgaris - bioraffinage - elektrisch veld - organellen
Microalgae are a potential feedstock for various products. At the moment, they are already used as feedstock for high-valuable products (e.g. aquaculture and pigments).
Microalgae pre-dominantly consist out of proteins, lipids and carbohydrates. This makes algae an interesting feedstock for various bulk-commodities. To successfully produce bulk-commodities, a multi-product biorefinery should be adopted that aims on production of both bulk- and high value co-products. In the downstream process, however, harvesting- and cell disruption are technological hurdles for cost effective multi-product biorefinery.
Flocculation is considered as a low-cost harvesting process. Flocculating microalgae at high salinities used to be not feasible We demonstrated that marine microalgae can successfully be flocculated and harvested by using cationic polymers.
In the second part of this thesis we studied Pulsed Electric Field (PEF) as potential cheap and non-disruptive technology to open microalgae. PEF-treatment evokes openings/’holes’ in micro-organisms. PEF in combination with a pre-treatment to weaken the cell wall resulted in release of proteins from microalgae at low energy consumption.
Recent advances in technology development learned that harvesting of micro-algae is no longer a bottleneck. Future research and development should focus on cell disruption and mild extraction technologies. Costs for the biorefinery will decrease by process simplification. For that unit operations for cell disruption and extraction need to be integrated.
This project was part of a large public private partnership program AlgaePARC biorefinery (www.AlgaePARC.com). Objective of this program is to develop a more sustainable and economically feasible microalgae production process. For that all biomass components (e.g. proteins, lipids, carbohydrates) should be used at minimal energy requirements and minimal costs while keeping the functionality of the different biomass components. Biorefining of microalgae is very important for the selective separation and use of the different functional biomass components.
Algen: onze oorspronkelijke omega-3 bron
Spruijt, J. - \ 2017
Wageningen University & Research - 6
algen - omega-3 vetzuren - voedseltechnologie - plantaardige oliën - algae - omega-3 fatty acids - food technology - plant oils
Brochure over de marktkansen voor omega-3 uit algen voor PUFAChain.
Toekomst AlgaePARC veiliggesteld
Barbosa, Maria - \ 2017
algae - algae culture - bioprocess engineering - biotechnology - experimental stations - research projects - test rigs - financing
AlgaePARC, de Wageningse proefaccommodatie voor algenonderzoek, kan voorlopig weer vooruit. Onderzoeker Maria Barbosa van Bioprocestechnologie haalde vier onderzoeksprojecten binnen, waaronder een groot EU-project.
From harmful to useful algae
Blaas, Harry - \ 2017
Wageningen University. Promotor(en): Carolien Kroeze. - Wageningen : Wageningen University - ISBN 9789463430357 - 117
algae - algae culture - adverse effects - nitrogen - phosphorus - rivers - eutrophication - waste water treatment - europe - algen - algenteelt - nadelige gevolgen - stikstof - fosfor - rivieren - eutrofiëring - afvalwaterbehandeling - europa
Eutrophication of coastal waters is a worldwide phenomenon. This study focuses on eutrophication in the coastal waters of Europe. Eutrophication is mainly a result of the increased transport of nutrients from watersheds by rivers to the coastal waters. Nutrient losses from watersheds are generally from agriculture, sewage, atmospheric deposition and from natural sources. In case of an overload of nutrients in the coastal waters, algal blooms may develop which increase the risk of hypoxia, fish mortality, and loss of biodiversity.
Algae can also be useful. They are increasingly considered an interesting product. For instance, micro-algae can be grow on land to produce proteins, lipids and fatty acids. Some studies indicate that micro-algae can be an important feedstock in the future for, for instance, the production of biodiesel. Moreover, macro-algae can be produced in seawater in sea farms. Macro-algae can be edible, or be used as a feedstock. By yielding macro-algae, nutrients are removed from the water, reducing coastal eutrophication.
The objective of this study is to analyse past and future trends in nutrient export by rivers to European seas with a focus on the role of algae. Three types of algae will be distinguished: (1) harmful algal blooms in coastal seas, (2) cultivation of micro-algae on land for the production of proteins, lipids and fatty acids, and (3) cultivation of multi cellular algae in seaweed farms for human consumption or other products.
To meet the objective the following research questions are addressed:
RQ1 To what extent do N and P loads exceed levels that minimize the risk of harmful algal blooms, and what are the relative shares of sources of N and P in rivers of the European Union?
RQ2 What are the potential consequences of large-scale land-based production of biodiesel from cultivated micro-algae in Europe for coastal eutrophication?
RQ3 Would it possible to cultivate and process micro-algae in a factory, and what is the environmental performance?
RQ4 To what extent can seaweed farming in combination with nutrient management in agriculture and waste water treatment reduce the potential for coastal eutrophication?
These questions are answered through model analyses. The Global NEWS (Nutrient Export from WaterSheds) model simulates river export of nutrients as function of human activities on land. It includes more than 6000 rivers worldwide. It can be used to quantify nutrient flows from land to sea for the years 1970, 2000, 2030 and 2050. For future years four scenarios have been implemented. One of these scenarios is named Global Orchestration and mostly used as a reference in this thesis. This scenario assumes a globalised world, with a reactive approach towards environmental problems. The model was released in 2010, has been validated for the years 1970 and 2000. The nutrients considered in the model are nitrogen (N) and phosphorus (P). In this thesis Global NEWS is used to calculate transport of nutrients to the coastal waters of Europe. The model uses ICEP (Indicator for Coastal Eutrophication Potential) values at the river mouths as an indicator for potentially harmful effects of nutrient enrichment. These ICEP values reflect the ratio of nitrogen and phosphorus to silica in coastal seas. A positive ICEP value indicates that nitrogen or phosphorus levels are too high, favouring conditions for potentially harmful algae to bloom.
In chapter 2 Global NEWS is used to calculate the transport of nutrients and ICEP values for 48 European rivers for the years 2000 and 2050. The model calculates a positive ICEP for 38 rivers in the year 2000, and for 34 rivers in the year 2050. This indicates that current policies are not so effective in reducing the river transport of nutrients. For polluted rivers the anthropogenic sources of the nutrients are investigated. For most rivers the dominant polluting sources are agriculture or sewage. The results indicate that a basin-specific policy is needed to reduce the risks of coastal eutrophication.
In chapter 3 the focus is on useful algae: micro-algae cultivation on land for, for instance, biodiesel production. The consequences of large-scale production of biodiesel on nutrient export by rivers to the European coastal waters are investigated. A scenario is developed assuming that a production of 0.4 billion m3 diesel from cultivated micro-algae. The cultivation is assumed to be in the open air, for instance in ponds or in closed tube systems. Such production levels would need a land surface area as large as Portugal. The Global NEWS model is used to calculate the amount of waste water from micro-algae production that will be transported to the coastal waters in this scenario. The results indicate that large-scale cultivation of micro-algae on land can become a source of nutrient pollution in rivers. In the scenario with large-scale micro-algae cultivation the future transport of nitrogen and phosphorus is considerably higher than in the reference scenario. To ensure sustainable production of biodiesel from micro-algae it is important to develop cultivation systems with low nutrient losses to the environment.
Chapter 4 presents a design of a factory for the cultivation and processing of micro-algae in an environmentally sound way. The factory does not use fossil fuels and applies maximum recycling of water and nutrients. In this factory it is possible to produce lipids, carbohydrates, proteins and minerals. The factory can be built on any piece of land, so there is no need to use arable land. The factory is independent of weather and climate. Energy can be delivered by wind mills. In this chapter an example of producing diesel in the factory is shown. In the 12 stories factory with a cultivation area of 1 hectare, 810 ton micro-algae can be cultivated per year. This is enough for the production of 386 ton diesel per year.
Chapter 5 focuses on mitigation of eutrophication in European coastal waters. A scenario is presented assuming different types of measures. The scenario first assumes that nutrient use efficiencies in agriculture are higher than today, and that waste water treatment in sewage systems is improved. In addition, it assumes that all excess N and P in coastal waters is harvested in seaweed farms producing edible macro-algae. In our scenario for 2050 there is seaweed farming in the coastal waters of 34 rivers mouths in Europe .NEWS The areas needed to ensure that ICEP values remain below 0 (low potential for coastal eutrophication) range between 0 and 952 km2 per river mouth.
This thesis shows that algae can be both harmful and useful. River export of nutrients can lead to coastal eutrophication increasing the risks of harmful algal blooms. On the other hand, micro-algae can be produced without environmental harm on land, and macro-algae can be useful in reducing pollution levels in coastal seas. This thesis could serve as a basis for environmental policies to stimulate the production of these useful algae. The methods to mitigate algal blooms and to use algae in a sustainable way in this thesis are also useful for other parts of the world.
Groen proceswater: zuivering brouwerijprocesafvalwater met microalgen
Dijk, W. van; Weide, R.Y. van der; Kroon, A. - \ 2016
Lelystad : ACRRES - Wageningen UR (PPO 721) - 42
brouwerij-industrie - brouwerijafvalwater - afvalwater - afvalwaterbehandeling - waterzuivering - algen - biomassa - biomassa productie - afvoerwater - flotatie - algenteelt - brewing industry - brewery effluent - waste water - waste water treatment - water treatment - algae - biomass - biomass production - effluents - flotation - algae culture
In 2012 is het project Groen Proceswater gestart. Hierin worden de mogelijkheden van zuivering van brouwerijprocesafvalwater met behulp van microalgen onderzocht. Dit is gedaan in een samenwerkingsverband van Heineken Nederland BV, Algae Food & Fuel en WUR-ACRRES. De resultaten behaald in 2012 en 2013 zijn beschreven in afzonderlijke rapporten. In dit rapport zijn de resultaten van 2014 en 2015 beschreven. In 2014 is onderzocht of een voorbehandeling van het proceswater via cavitatie-flotatie het zuiveringsresultaat kan verbeteren en in 2015 of de energie-input van de LEDbelichting kan worden verminderd door te flashen (afwisselende licht-donker periodes op microseconde tijdschaal). Verder is in 2015 en 2016 via een literatuurstudie gekeken naar de verwaarding van de geproduceerde algenbiomassa en zijn de conclusies van het gehele project samengevat.
Biobased Economy: Algen, het groene goud
Barbosa, M.J. - \ 2016
Wageningen : Wageningen University & Research
algenteelt - algen - biomassa productie - teeltsystemen - cultuurmethoden - biobased economy - algae culture - algae - biomass production - cropping systems - cultural methods - biobased economy
De wereldbevolking groeit en daarmee ook de behoefte aan energie, chemicaliën, en producten zoals plastic, zeep, schoonmaakmiddelen, verf en lijm. Om daarin te kunnen voorzien, moeten we overstappen naar een economie, waarin we de grondstoffen voor deze producten uit duurzame, hernieuwbare bronnen halen. Een voorbeeld van een van de meest duurzame oplossingen zijn micro-algen. Maria Barbosa van Wageningen University & Research vertelt er alles over.
Mild disintegration of green microalgae and macroalgae
Postma, Richard - \ 2016
Wageningen University. Promotor(en): Michel Eppink; Rene Wijffels, co-promotor(en): Giuseppe Olivieri. - Wageningen : Wageningen University - ISBN 9789462579477 - 181
algae - chlorella vulgaris - bioprocess engineering - biorefinery - proteins - milling - carbohydrates - biobased economy - disintegrators - technology - extraction - algen - chlorella vulgaris - bioproceskunde - bioraffinage - eiwitten - maling - koolhydraten - biobased economy - desintegrators - technologie - extractie
An increased worldwide protein demand for food and feed and the necessity to release the water soluble proteins in the first stage of the cascade biorefinery require the development of mild protein extraction technologies. Cell disintegration is the first hurdle and is considered as one of the most energy consuming steps. Therefore, this thesis focused on the development of a mild, scalable and energy efficient disintegration technology for green microalgae and macroalgae (seaweed) aimed on extraction of water soluble components (like proteins and carbohydrates).
For microalgae disintegration, two main technologies were investigated. First of all the conventional technology bead milling and second a novel approach using Pulsed Electric Field (PEF). In Chapter 2 a benchmark was set by means of bead milling for the release of water soluble protein from the green microalgae Chlorella vulgaris. Overall, protein yields between 32 and 42% were achieved, while the energy consumption was reduced with 85% by selective protein extraction to values as low as 0.81 kWh kgDW-1. Remarkably, the benchmark was much better than expected.
In Chapter 3 the bead mill was further optimized by decreasing the applied bead size, furthermore the applicability of bead milling on two additional microalgae species (Neochloris oleoabundans, Tetraselmis suecica) was shown. In addition, to be able to better understand the disintegration mechanism, the so-called stress model was applied. This model describes the comminution process in a bead mill as function of the amount of bead contacts and the force of each impact. The release kinetics could be improved and thereby the specific energy consumption could be reduced to 0.45‒0.47 kWh kgDW-1 by using 0.3 mm beads for all algae.
Chapter 4 describes a screening on the applicability of PEF, over a broad range of operating conditions, for the extraction of water soluble proteins from the microalgae C. vulgaris and N. oleoabundans. No substantial protein yields were observed under the investigated conditions. This led to the conclusion that PEF is not suitable to release water soluble proteins, not even at specific energy consumptions much higher than those for the benchmark, bead milling.
In Chapter 5 it was attempted to improve the performance of PEF by investigating the synergistic effect with the processing temperature. The PEF experiments were performed using a pilot scale continuous flow electroporation unit in which the processing temperature was controlled between 25 – 65 °C. The results showed that under the tested conditions, the combined PEF-Temperature treatment did not cause substantial disintegration of the algal cells to effectively release water soluble proteins.
In addition to the microalgae, macroalgae were subject of investigation in the search for new protein sources in Chapter 6. Four batch technologies were used to disintegrate the green macroalgae Ulva lactuca, being; osmotic shock, enzyme incubation, PEF and High Shear Homogenization (HSH). In descending order the highest protein yields per treatment; HSH (~40%) > enzyme degradation (~25%) > osmotic shock (~20%) > PEF (~15%).
In the final chapter the main results and remaining bottlenecks are discussed and a future outlook on microalgae disintegration is presented. To date, bead milling is the only technology able to disintegrate fresh microalgae at specific energy consumptions below 10% of the total energy available from the algae and release substantial amounts of water soluble protein. The future outlook was based on a techno-economic evaluation, which showed that the cultivation costs are limiting the economic feasibility of microalgae biorefinery. Future focus should be on the cultivation.
Ionic liquid pre-treatment of microalgae and extraction of biomolecules
Desai, Rupali K. - \ 2016
Wageningen University. Promotor(en): Michel Eppink; Rene Wijffels. - Wageningen : Wageningen University - ISBN 9789462579804 - 126
salts - liquids - fractionation - extraction - hydrophobicity - algae - biomass production - zouten - vloeistoffen (liquids) - fractionering - extractie - hydrofobiciteit - algen - biomassa productie
Liquid-liquid extraction (LLE) techniques are widely used in separation primarily due to ease of scale up. Conventional (LLE) systems based on organic solvents are not suitable for extraction of fragile molecules such as proteins as it would result in denaturation. On the other hand aqueous biphasic system though suitable for extraction of proteins they are restricted by limited polarity range. Ionic liquids are salts which are liquid at room temperature. Ionic liquids have gained interest in extraction over the past years due to its non-volatility and tunable property. In this thesis we explored the feasibility of using two ionic liquid based systems for extraction: 1) Ionic liquid based aqueous two phase system for extraction of microalgae proteins and 2) ionic liquid based emulsions for separation of hydrophilic (e.g. proteins) and hydrophobic (e.g. pigments) components from complex biomass such as microalgae. Additionally the influence of IL pre-treatment on microalgae cell walls and subsequent fractionation of its components (e.g. proteins, pigments, lipids) was also investigated.
The fatter the better : selecting microalgae cells for outdoor lipid production
Dominguez Teles, I. - \ 2016
Wageningen University. Promotor(en): Rene Wijffels, co-promotor(en): Maria Barbosa; Dorinde Kleinegris. - Wageningen : Wageningen University - ISBN 9789462578821 - 164
algae - chlorococcum - lipids - lipogenesis - fat - production - phenotypes - inoculum - diameter - cells - sorting - algen - chlorococcum - lipiden - lipogenese - vet - productie - fenotypen - entstof - diameter - cellen - sorteren
In chapter 1 we introduce microalgae, photosynthetic microorganisms with potential to replace commodities (such as food, feed, chemicals and fuels). Production costs are still high, reason why microalgae are still only economically feasible for niche markets. We suggest to borrow the concept of plant domestication to select industrial microalgae cells. Two approaches can be successfully used to domesticate microalgae: adaptive laboratory evolution (ALE) and fluorescence assisted cell sorting (FACS). ALE takes advantage of the natural adaptability of microorganisms to different environments, while FACS actually select cells with specific phenotypes. This thesis aimed to select cells of Chlorococcum littorale with improved phenotypes, assuming that these cells could establish new populations with increased industrial performance.
In Chapter 2 we wanted to know what happened during time to biomass and lipid productivities of Chlorococcum littorale repeatedly subjected to N-starvation. We tested 2 different cycles of N-starvation, short (6 days) and long (12 days). Short cycles didn’t affect lipid productivity, highlighting the potential of C. littorale to be produced in semi-continuous cultivation. Repeated cycles of N-starvation could have caused adaptations of the strain. Hence, we also discussed the implications of using repeated N-starvation for adaptive laboratory evolution (ALE) experiments. Chapter 3 introduces a method to detect and to select microalgae cells with increased lipid content. The method requires only the fluorescence dye Bodipy505/515 dissolved in ethanol, and the method was designed to maintain cellular viability so the cells could be used to produce new inoculum. In chapter 4 we evaluated a question that emerged while deciding which criteria to use to sort lipid-rich cells: does cellular size affects lipid productivity of C. littorale? We hypothesized that cells with different diameters have different division rates, which could affect lipid productivity. Therefore, we assessed the influence of cell diameter, as a sorting parameter, on both biomass and lipid productivity of Chlorococcum littorale (comparing populations before and after sorting, based on different diameters). Results showed that the size of vegetative cells doesn’t affect the lipid productivity of C. littorale. In chapter 5 we present a strategy to sort cells of C. littorale with increased TAG productivity using the method developed at chapter 3. Both the original and the sorted population with the highest lipid productivity (namely, S5) were compared under simulated Dutch summer conditions. The results confirmed our data from experiments done under continuous light: S5 showed a double TAG productivity. Our results showed also that the selected phenotype was stable (1.5 year after sorting) and with potential to be used under industrial conditions. In chapter 6 we extrapolated our results (indoor and outdoor) to other climate conditions. We ran simulations changing the light conditions to four different locations worldwide (the Netherlands, Norway, Brazil and Spain) to estimate both biomass and TAG productivities. Results indicated that biomass yields were reduced at locations with higher light intensities (Brazil/Spain) when compared with locations with lower light intensities (Norway/Netherlands). Hence, the choice of location should not be based on light intensity, but on how stable irradiation is. Chapter 7 is the general discussion of the thesis, demonstrating that both ALE and FACS are effective approaches to select industrial microalgae cells. We also present our view on how ALE and FACS could further improve microalgae strains for industry.