Fernandes, F.; Silkina, A.; Gayo-Peláez, J.I.; Kapoore, R.V.; de la Broise, D.; Llewellyn, C.A. 2022

Fernandes, F.; Silkina, A.; Gayo-Peláez, J.I.; Kapoore, R.V.; de la Broise, D.; Llewellyn, C.A. Microalgae Cultivation on Nutrient Rich Digestate: The Importance of Strain and Digestate Tailoring under PH Control. Appl. Sci. 2022, 12, 5429.


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The bioremediation of digestate using microalgae presents a solution to the current eutroph-ication issue in Northwest Europe, where the use of digestate as soil fertiliser is limited, thus resultingin an excess of digestate. Ammonium is the main nutrient of interest in digestate for microalgalcultivation, and improving its availability and consequent uptake is crucial for optimal bioreme-diation. This work aimed to determine the influence of pH on ammonium availability in culturesof two green microalgae, additionally screened for their growth performances on three digestatesproduced from different feedstocks, demonstrating the importance of tailoring a microalgal strainand digestate for bioremediation purposes. Results showed that an acidic pH of 6–6.5 resulted in abetter ammonium availability in the digestate media, translated into better growth yields for bothS. obliquus(GR:0.099±0.001 day−1; DW: 0.23±0.02 g L−1) andC. vulgaris(GR: 0.09±0.001 day−1;DW:0.49±0.012 g L−1). This result was especially true when considering larger-scale applicationswhere ammonium loss via evaporation should be avoided. The results also demonstrated thatdigestates from different feedstocks resulted in different growth yields and biomass composition,especially fatty acids, for which, a digestate produced from pig manure resulted in acid contentsof 6.94±0.033% DW and 4.91±0.3% DW inS. obliquusandC. vulgaris, respectively. Finally, thiswork demonstrated that the acclimation of microalgae to novel nutrient sources should be carefullyconsidered, as it could convey significant advantages in terms of biomass composition, especially fattyacids and carbohydrate, for which, this study also demonstrated the importance of harvesting time.


Keywords:digestate; microalgae; pH control; ammonium; bioremediation

Elshamy, M., Rösch, C. 2022

Elshamy, M., Rösch, C. Animal Feed from Microalgae Grown on Biogas Digestate as Sustainable Alternative to Imported Soybean Meal. Bioenerg. Res. (2022).


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This work investigates microalgae’s potential to reduce North-Western Europe environmental burden from excess nitrogen polluting groundwater and provide an environmental replacement for current animal feed. Algae uptake nutrients from nutrient-rich digestate from biogas plants and turn them into high-value feedstock, which can substitute soybean meal imported from overseas and avoid deforestation of rainforests. Life cycle assessments were conducted based on novel and original data from engineers in academia and industry, acquired through pilot-scale research facilities in the UK, France, and Belgium. The findings of this study highlight the environmental impacts of three different technologies with varying scales of production. Mixotrophic algal feed production shows higher productivities and fewer energy demands than the phototrophic systems. Still, the process is less favourable due to higher impacts on human health and ecosystems than imported soybean meal, whereas the impact on resources is more negligible. Sensitive analysis indicates that further technological development, increased productivities, and renewable energy use can improve the environmental competitiveness of algal feedstock. Since this process is a promising solution for making feedstock out of excess nutrients and having an alternative source for imported soybean meal with a comparable, efforts to realize these improvements and successfully upscaling algal feedstock production in combination with biogas plants are justified and can improve the environmental footprint of algal biomass production.

Jai Sankar Seelam et al, 2022

Jai Sankar Seelam, Marcella Fernandes de Souza, Peter Chaerle, Bernard Willems, Evi Michels, Wim Vyverman, Erik Meers

Maximizing nutrient recycling from digestate for production of protein-rich microalgae for animal feed application,
Volume 290,
ISSN 0045-6535,


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Abstract: The integration of phototrophic microalgal production and anaerobic digestion can recycle excess nutrients across European surplus hotspots to produce protein-rich biomass for nutritional applications. However, the challenging physico-chemical properties of raw digestate constrain microalgal growth and limit digestate valorization potential. This study focused on the pre-treatment of food waste-based digestate using paper-filtration to improve its properties for cultivating Desmodesmus sp. and Chlorella vulgaris. The microalgal growth performance in paper-filtered digestate (PFD, 10 μm-pore size) was then compared to growth in membrane-filtered digestate (MFD, 0.2 μm-pore size). A microplate-based screening coupled with Cytation device assessment of PFD and MFD samples after dilution and with/without phosphorus supplementation showed that PFD was the best substrate. Moreover, phosphorus supplementation resulted in improved growth at higher digestate concentrations (5–10% v/v PFD), indicating the importance of using a balanced growth medium to increase the volumetric usage of digestate. Results were validated in a 3-L bioreactor at 10% PFD with phosphorus supplementation, reaching a biomass concentration of 2.4 g L−1 with a protein and carbohydrate content of 67% and 13% w/w respectively. This trial indicates that paper-filtration is a promising pre-treatment technique to maximize digestate recycling and deliver a sustainable animal feed-grade protein alternative.
Keywords: Animal feed; Digestate; Microalgae; Microplate experiments; Paper-filtration; Photobioreactor





Claudio Fuentes-Grünewald et al, 2021

Claudio Fuentes-Grünewald, José Ignacio Gayo-Peláez, Vanessa Ndovela, Eleanor Wood, Rahul Vijay Kapoore, Carole Anne Llewellyn,

Towards a circular economy: A novel microalgal two-step growth approach to treat excess nutrients from digestate and to produce biomass for animal feed
Bioresource Technology,
Volume 320, Part A,
ISSN 0960-8524,


Link to the paper 
Abstract: Implementing a circular economy aimed at reusing resources is becoming increasingly important for industry. Microalgae fit within a circular economy by being able to bioremediate nutrient waste and as a source of biomass for several commercial applications. Here, we report a novel validation of a circular economy concept using microalgae at a relevant industrial scale with a new two-phase process. During the first phase, biomass was grown autotrophically, biomass was then concentrated using membrane technology for the second phase where mixotrophic conditions were applied to boost growth further. Microalgae cultures were able to grow (13.8 g/L), uptake and bioremediate nutrients (Nitrogen > 134 mg/L/day) from an anaerobic digestion side-stream (digestate), obtaining high-quality microalgae biomass (>45% protein content) suitable for use as animal feed, closing the circular economy loop for industrial applications.
Keywords: Circular economy; Microalgae; Digestate; Membrane filtration

Rahul Vijay Kapoore et al, 2021

Rahul Vijay Kapoore, Eleanor E. Wood, Carole A. Llewellyn,

Algae biostimulants: A critical look at microalgal biostimulants for sustainable agricultural practices,

Biotechnology Advances,
Volume 49,
ISSN 0734-9750,

link to the paper 

Abstract: For the growing human population to be sustained during present climatic changes, enhanced quality and quantity of crops are essential to enable food security worldwide. The current consensus is that we need to make a transition from a petroleum-based to a bio-based economy via the development of a sustainable circular economy and biorefinery approaches. Both macroalgae (seaweeds) and microalgae have been long considered a rich source of plant biostimulants with an attractive business opportunity in agronomy and agro-industries. To date, macroalgae biostimulants have been well explored. In contrast, microalgal biostimulants whilst known to have positive effects on development, growth and yields of crops, their commercial implementation is constrained by lack of research and cost of production. The present review highlights the current knowledge on potential biostimulatory compounds, key sources and their quantitative information from algae. Specifically, we provide an overview on the prospects of microalgal biostimulants to advance crop production and quality. Key aspects such as specific biostimulant effects caused by extracts of microalgae, feasibility and potential of co-cultures and later co-application with other biostimulants/biofertilizers are highlighted. An overview of the current knowledge, recent advances and achievements on extraction techniques, application type, application timing, current market and regulatory aspects are also discussed. Moreover, aspects involved in circular economy and biorefinery approaches are also covered, such as: integration of waste resources and implementation of high-throughput phenotyping and -omics tools in isolating novel strains, exploring synergistic interactions and illustrating the underlying mode of microalgal biostimulant action. Overall, this review highlights the current and future potential of microalgal biostimulants, algal biochemical components behind these traits and finally bottlenecks and prospects involved in the successful commercialisation of microalgal biostimulants for sustainable agricultural practices.

Keywords: Algae biostimulants; Sustainable agriculture; Microalgae biotechnology; Metabolomics; Consortia; Biorefinery; Circular economy; Bioremediation

Fleuriane Fernandes et al, 2020

Fleuriane Fernandes, Alla Silkina, Claudio Fuentes-Grünewald, Eleanor E. Wood, Vanessa L.S. Ndovela, Darren L. Oatley-Radcliffe, Robert W. Lovitt, Carole A. Llewellyn,

Valorising nutrient-rich digestate: Dilution, settlement and membrane filtration processing for optimisation as a waste-based media for microalgal cultivation,
Waste Management,
Volume 118,
Pages 197-208,
ISSN 0956-053X,

Link to the paper  

Abstract: Digestate produced from the anaerobic digestion of food and farm waste is primarily returned to land as a biofertiliser for crops, with its potential to generate value through alternative processing methods at present under explored. In this work, valorisation of a digestate resulting from the treatment of kitchen and food waste was investigated, using dilution, settlement and membrane processing technology. Processed digestate was subsequently tested as a nutrient source for the cultivation of Chlorella vulgaris, up to pilot-scale (800L). Dilution of digestate down to 2.5% increased settlement rate and induced release of valuable compounds for fertiliser usage such as nitrogen and phosphorus. Settlement, as a partial processing of digestate offered a physical separation of liquid and solid fractions at a low cost. Membrane filtration demonstrated efficient segregation of nutrients, with micro-filtration recovering 92.38% of phosphorus and the combination of micro-filtration, ultra-filtration, and nano-filtration recovering a total of 94.35% of nitrogen from digestate. Nano-filtered and micro-filtered digestates at low concentrations were suitable substrates to support growth of Chlorella vulgaris. At pilot-scale, the microalgae grew successfully for 28 days with a maximum growth rate of 0.62 day−1 and dry weight of 0.86  g⋅L−1. Decline in culture growth beyond 28 days was presumably linked to ammonium and heavy metal accumulation in the cultivation medium. Processed digestate provided a suitable nutrient source for successful microalgal cultivation at pilot-scale, evidencing potential to convert excess nutrients into biomass, generating value from excess digestate and providing additional markets to the anaerobic digestion sector.
Keywords: Digestate; Membrane filtration; Settlement and dilution; Microalgae; Chlorella vulgaris; Pilot-scale

Carole A. Llewellyn et al, 2020

Carole A. Llewellyn, Rahul Vijay Kapoore, Robert W. Lovitt, Carolyn Greig, Claudio Fuentes-Grünewald, Bethan Kultschar

Llewellyn C.A., Kapoore R.V., Lovitt R.W., Greig C., Fuentes-Grünewald C., Kultschar B. (2019) Deriving Economic Value from Metabolites in Cyanobacteria. In: Hallmann A., Rampelotto P. (eds) Grand Challenges in Algae Biotechnology. Grand Challenges in Biology and Biotechnology. Springer, Cham.

Part of the Grand Challenges in Biology and Biotechnology book series (GCBB)

Abstract: This chapter focuses on the challenges associated with achieving economic value from metabolites derived from cyanobacteria. Significant advances have been made in cyanobacterial biotechnology in the last few years. However, the field is still immature, and many challenges remain. We start with a critical overview of the main technologies associated with cultivation, cell disruption and metabolite extraction. Then, we provide an overview of current significant metabolite groups from cyanobacteria relevant to industry covering phycobilins, carotenoids, polysaccharides, peptides, lipids, mycosporine-like amino acids, polyhydroxyalkanoates, cyanotoxins and platform chemicals, and the potential for stable isotopes production. We cover metabolites that are already in the market and those with future potential with a focus on spirulina (Arthrospira) the most commercially developed species of cyanobacteria. As large-scale cultivation and downstream processing techniques continue to develop further, combining this with a systems biology and biorefinery approach will ensure that the best economic and environmental sustainability value can be achieved.


Stiles A.V. William et al, 2018

William A.V. Stiles, David Styles, Stephen P. Chapman, Sandra Esteves, Angela Bywater, Lynsey Melville, Alla Silkina, Ingrid Lupatsch, Claudio Fuentes Grünewald, Robert Lovitt, Tom Chaloner, Andy Bull, Chris Morris, Carole A. Llewellyn,

Using microalgae in the circular economy to valorise anaerobic digestate: challenges and opportunities,
Bioresource Technology,

Volume 267,
Pages 732-742,
ISSN 0960-8524,


 Link to the paper 


Abstract: Managing organic waste streams is a major challenge for the agricultural industry. Anaerobic digestion (AD) of organic wastes is a preferred option in the waste management hierarchy, as this process can generate renewable energy, reduce emissions from waste storage, and produce fertiliser material. However, Nitrate Vulnerable Zone legislation and seasonal restrictions can limit the use of digestate on agricultural land. In this paper, we demonstrate the potential of cultivating microalgae on digestate as a feedstock, either directly after dilution, or indirectly from effluent remaining after biofertiliser extraction. Resultant microalgal biomass can then be used to produce livestock feed, biofuel or for higher value bio-products. The approach could mitigate for possible regional excesses, and substitute conventional high-impact products with bio-resources, enhancing sustainability within a circular economy. Recycling nutrients from digestate with algal technology is at an early stage. We present and discuss challenges and opportunities associated with developing this new technology.

Keywords: Anaerobic digestion; Algae; Nutrient recycling; Livestock feed; Circular economy

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