Final Report Summary - BIOALGAESORB (Enabling European SMEs to remediate wastes, reduce GHG emissions and produce biofuels via microalgae cultivation) Executive Summary:The overall cooperation between partners in the project has been very good and the partners have been actively involved in the project. There has been good communication between the partners at both formal and informal meetings, phone calls, email, etc. The participants have been very active with regards to using the project web established at Basecamp. Through this site, all participants have been able to monitor ongoing project progress and access all meeting minutes, presentations and reports. During Reporting Period 3, the following Management Board meetings are held in the BioAlgaeSorb project:• 30 Month meeting: 29th November 2012. Swansea, UK. Hosted by Swansea University• 36 Month meeting: 23th of May 2013, Oslo, Norway. Hosted by Norsk Bioenergiforening and Teknologisk Institutt AS.Agendas are prepared and invitation to the Management meetings is distributed to beneficiaries minimum 1 month prior to the meeting. Minutes of meetings are prepared and made available for all participants at project web hotel. In order to save costs and travel expenses, Technical meetings are held in association with the Management Board meetings. The main objective of the Technical meetings are to discuss the project results and RTD activity in further detail and especially plan and coordinate the RTD activity for the next 3-6 months period. Separate agenda and minutes are prepared for the Technical meetings. In addition to the Management Board meetings and the associated Technical meetings listed, a very high number of less formal technical meetings have been arranged to discuss the technical aspects of the project. These meetings do included physical meetings between a limited no. of beneficiaries and telephone meetings.Scientific and Technical activity in Reporting Period 3 do especially focus on WP2 to 6 (RTD activities). In addition, considerable activity and progress is made in WP7 Demonstration activities and WP 8 IPR, Training and Dissemination. Major deliverables are successfully completed Reporting Period 3 include:• Month 30 – Deliverable D2.3: Report on the physical and chemical properties of microalgae concentrates and their drying characteristics and feasibility of novel microalgae harvesting processes, electro-flocculation and ozonated air floatation• Month 30 - Deliverable D4.1: Report on pyrolysis of microalgal fraction using heterogeneous catalysts. This delivery was delayed in Reporting Period 2 due to delayed delivery of required equipment from the supplier. Unfortunately, this delay was not foreseen and came quite surprisingly for CREAR who is responsible for this activity. A draft delivery report of D4.1 was provided in Reporting Period 2. An updated delivery report is submitted in Reporting Period 3. The delay with regards to D4.1 did not represent any delay in the overall progress of the project.• Month 30 - Deliverable D4.2: Report on transesterification and decarboxylation of microalgal fractions using heterogeneous catalysts• Month 36 – Deliverable D5.2: Report on the result of bio-oil production and analysis and bio-oil in prime movers• Month 30 – Deliverable D6.1: Report describing the work performed in WP6, including the building of prototype(s), system integration, functional testing and benchmarking• Month 36 – Deliverable D6.3: Final report on risk assessment and contingency plan• Month 36 – Deliverable D6.4: Report on economic assessment and viability of different process routes evaluated.• Month 34 – Deliverable D7.1: Report on results from the demonstration/ case study• Month 36 - Deliverable D8.3: Final report on potentially competitive patents and a plan for patent application(s) if required with exploitation agreements between partners• Month 36 – Deliverable D8.4: Report on training material and evaluation from the participants• Month 36 – Deliverable D8.6: Dissemination and Use Plan – final• Month 36 – Deliverable D8.7: Report on legal and societal aspects and implications of the projectIn addition, in the reporting period 3 the following milestones are completed:• Milestone no. 6:” Chemical conversion processes developed for biofuel production from microalgae lipid fraction”.• Milestone no. 7 “Pilot reactor ready for testing” is partially completed.• Milestone no. 8: “First repeatable production of bio-oil (few litres)” – partly achievedProject Context and Objectives:This project addresses specific needs of several key European SME groupings, via integration of technologies for effluent water remediation and the production and exploitation of microalgae biomass for mitigation of climate change (renewable energy generation, carbon dioxide (CO2) capture, utilisation of organic wastes) and the production of valuable bio-products using a biorefinery approach. The overall aims of the project are to: • Increase knowledge on the bioconversion of industrial and agricultural/aquacultural effluents to microalgae biomass, as a sustainable raw material for biofuels and other value added applications;• Provide new carbon neutral fuel sources for biomass power plants and biodiesel manufacture;• Provide new sources of sustainable and carbon neutral high quality fine chemicals extracted from microalgae biomass;• Reduce the discharge of CO2 to the atmosphere from biomass and fossil fuel power plants and other industrial processes reliant on combustion of fossil fuels;• Reduce the nutrient loading of effluent waters from livestock production systems which are responsible for the largest proportion of organic waste in Europe, including most ammonia emissions. • Increase know how and competence within a range of European SME-dominated industries.Scientific objectives include:• Optimise parameters for the rapid growth of (especially carbon-?tolerant?) microalgae, using waste-water nutrients and/or CO2-rich industrial flue gases (up to 70% concentration in growth media), to high densities and in scalable cultivation systems• Develop efficient and reliable microalgae harvesting and dewatering processes;• Develop effective processes (up to 50% oil by weight) for the conversion of dewatered microalgal biomass into biofuels and/or directly into energy;• Optimise physical and chemical fractionation and transformation methods for those biomass components not directly converted to biofuels or energy;Technological objectives include:• Assess the viability of the new processes and products developed, incorporating coupled process and financial models;• Develop an industry-based model to assess a variety of strategies that maximise the value of the microalgal biomass in a changing market environment. The work plan is structured in the following Work Packages (type of activity in brackets):• WP 1 - Scientific Understanding and Advancement of Operational parameters (RTD)• WP2 – Optimization of MicroAlgae Cultivation on Effluents (RTD)• WP3 – MicroAlgae Harvesting Technologies (RTD)• WP4 – Refinery and Upgrading Process for MicroAlgae Biomass (RTD)• WP5 – Termo-chemical Conversions of Microalgae Biomass (RTD)• WP6 – System integration and Industrial validation (RTD)• WP7 - Demonstration (DEMO)• WP8 – IPR, Training and Dissemination (OTHER)• WP9 - Consortium Management (MANAGEMENT) Project Results:Work package 2 – Optimization of Microalgae Cultivation on EffluentsWP 2 – Objectives• To develop processes for pre-treating and re-formulating aqueous and gaseous effluents as effective nutrient sources for cultivated microalgae;• To select suitable microalgae species for mass cultivation on representative effluent sources; • To develop robust methods for cultivating the selected microalgae species on representative effluent sources; • To assess the technical feasibility of the developed processes for microalgae mass cultivation at SME locations.In reporting period 3, activity and progress include the following tasks:• Task 2.3 Define Bioreactor and Raceway Operating Conditions for Selected Microalgae Species, Effluent Sources and Operating Locations• Task 2.4 Control of Nutrient Delivery and Optimisation of Transfers in Microalgae Cultivation Systems• Task 2.5 Modelling Approaches for Technical Evaluations• Task 2.6 Prototype Development and TestingWorkPackage 2 is successfully completed including Deliverable D2.3.Task 2.3 Define Bioreactor and Raceway Operating Conditions for Selected Microalgae Species, Effluent Sources and Operating LocationsAs it was decided at the project meeting in Swansea, two supplemental trials were run within the frame of task 2.3. In the first experiment Chlorella minutissima was cultured in a bioreactor where flue gas was the main carbon source. In a second trial, Nannochloropsis sp. was cultured in a bioreactor where artificial media from industrial wastes were used as nutrients and flue gases were added as well. The biomass (1 kg) was harvested and sent to partner CREAR in Italy.A large-scale experiment was run at the facilities of HCMR where a tubular bioreactor of 900 L was run at semi-continuous mode using mainly flue gases. Flue gases produced by an oil heater were added to the culture continuously together with air through an air-lift mode to provoke a circulation of the water mass in the bioreactor at the same time. In addition, the acidity of the culture was monitored by a pH sensor and pure CO2 was added when necessary. This means that pure CO2 gas was added when the amount of CO2 supplied by the flue gases was not sufficient to decrease the pH below a set value (7.2). As this experiment was run in middle of the winter (5th of December to 5th of January 2013), the amount of flue gas was high due to increased usage of the oil heater and it was the main carbon source for the culture. This bioreactor was a modified version from bioreactor used in previous trials for the Bioalgaesorb project, as no mechanical pump and no dark-phase was used and it was a system based on air-lift from air added though hoses at about 50 cm from the water surface. The total culture volume was about 900 L where is consisted almost in its whole of a transparent phase (photophase). It should be mentioned here that there was no cooling unit connected to this device. In this sense it was a system adapted to perform better on winter time. This is also the reason for the timing of these experiments in winter/early spring period (December-March). The biomass of Chorella minutissima during the experimental period was about 120-140 millions cells per mL, and in terms of packed cell volume it reached about 1.5 mL/L of culture.Satisfactory growth was obtained this experiment by use of flue gases. A total of 1900 liters of algae culture were harvested. As the biomass was about 1.5 g per L , this means that the biomass harvested was: 1900 liters × 1.5 g = 2850 g wet weight of microalgae biomass. Dry weight is about 15% of the wet biomass so it estimated to about 427 DW. The measurements of physiological state of photosynthetic mechanism by use of photo fluorescence (PEA-Photosynthetic Efficiency analyzer) using the Fv/Fm index showed that cells showed an optimal state of the photosynthetic mechanism. The index ideally should be between 0.6 and 0.8. Light and temperature as expected was quite variable in the middle of winter, but nevertheless sufficient to support microalgae growth. The use of flue gas had no negative impact on the microalgae cells which showed a satisfactory growth. Use of microalgae biomass for example in aquaculture purposes should however be evaluated in other terms, such as content of heavy metals or increased concentration of other contaminants present in the flue gases. Nevertheless, also this hinder could be overcome by use of a water trap or other filter before injection of the flue gases in the microalgae culture.The bioreactor used was similar to the one described in the experiment above only smaller (300 L). The differences were that (1) Nannochloropsis sp. was used as the cultured organism and (2) that the culture was initiated for the specific experiment. An adaptive strategy was used so the artificial media, so the culture at inoculation was added with only 25% of the total media in the form of experimental and the rest as commercial F/2. This percentage was gradually increased to 100%. The experiment was initiated the 18th of February and ended the 4th of March. The biomass harvested was centrifuged by use of filtration rig and harvested biomass was sent to Florence. The biomass in terms of cell density was about 80 millions cells/mL, and in terms of packed cell volume about 2 mL/L. Nannochloropsis cells are larger than Chlorella minutissima and culture of Nannochloropsis reach in general lower cell densities than Chlorella minutissima. In this second trial the gradual adaptation of the algae to the experimental media allowed the satisfactory culture of the algae for about two weeks. No need to adjustment of pH was shown. In a previous preliminary trial, run with the same artificial growth media, the microalgae collapsed 4-5 days after inoculation. In this first trial, experimental medium prepared at SU was used at a 100% strength from the day of inoculation, whereas in this trial a gradual adaptation from classical F/2 medium to experimental growth medium was done and the culture lasted as long as this experimental medium was available. The experiment was thus terminated after two weeks because the amount of experimental medium was limited (30 L). No problem with pH was shown. Acidity was carefully monitored during inoculation and no important decrease of pH was shown. This however, could be a problem in culture of freshwater microalgae. In total 400 liters were harvested. This contained about 800 g of wet biomass. A total biomass of 1 kg dry weight of Nannochloropsis produced with the same bioreactor was shipped to CREAR.Task 2.4 Control of Nutrient Delivery and Optimisation of Transfers in Microalgae Cultivation SystemsThe objective is to quantify gas mass transfers in order to identify potential problems of CO2 depletion/O2 super saturation and to resolve these through different gas injection and degassing options. SO, the main purpose with the automation system is to follow up the concentration of CO2/O2 gas in the water, PH value, Turbidity, Pressure. This part of the BioAlgeSorb project consisted of making an instrumentation and integration plan based on a system that provides sufficient information from the sensors for water quality.Sensor selections are based on the Instrumentation list generated from the Process and instrumentation diagram (P&ID). Selections of other electrical equipment are based on the control cabinet layout and the standard PLC control system setup which was used before as a basis for the several control system projects.PIR 7200 proof infrared gas detector for continuous monitoring of carbon dioxide was selected by TI. The gas transmitters are designed for use under the most adverse conditions. Robust stainless steel housing protects the electronic and the optical components.The Saia PCD3 PLC system was chosen as main control unit for BioAlgeSorb project. This is a standard, mainstream PLC with a wide range of standard and specified input and output modules which can be used with the PLC unit. The unit can be used for all kinds of controlling purposes, from the simplest one to the most advanced. It has numerous variations of effective communication possibilities and can easily be fitted to all kinds of systems and networks. All Saia PLC units use the same PG5 programming platform which is very convenient when one or more operators need to change the PLC program from different locations. Process value limits are set by the operator and are constantly controlled by program loops. If high or low limits are exceeded, error bit is triggered. High and low alarms are used as safety measures in the program loops during the process.Task 2.5 Modelling Approaches for Technical EvaluationsDynamic models will be constructed of each component part of the microalgae production system, building on information obtained from Task 1.4 and Tasks 2.1-3 for the biological and physical processes involved. The purpose is to integrate information from other components of WP2 into a dynamic testable platform for subsequent system integration and industrial validation activities (WP6).The computer models will make use of mechanistic descriptions of microalgal physiology to ensure that the description of the biological behaviour takes appropriate account of changes in physical parameters including light, water turbidity, nutrient types and concentrations, temperature and pH. These biological descriptions will be placed within descriptions of commercial PBRs and raceways of different size, shape, dilution, illumination, temperature, nutrient influx etc., i.e. factors that affect the ability of the microalgae to grow and remove nutrients. Combinations of different PBR and raceway configurations will be tested using these models. Coupled with these descriptors will be models describing the fate of material harvested from the reactors, especially with respect to the recycling of water and nutrients.Both the biological and physical descriptors will be flexible, enabling the examination of different scenarios both with respect to system configurations within a given geographic zone, and also between geographic zones. This is important given significant differences across Europe with respect to light, water supply, nutrient availability, land and energy costs, etc.The ability of the model to fit experimental data was tested in a growth trial in a Varicon biofence installation at Swansea.At SU, elemental analysis of algae samples was carried out on samples from growth experiments at HCMR, IngrePro and SU (Tasks 2.2 and 2.3). The obtained carbon and nitrogen cell quota data contributed to model validation for simulations at different latitudes.The model predicted data showed a close fit to the measured data obtained by a growth experiment with Nannochloropsis sp. in a Varicon biofence at Swansea. The development of C-Biomass, N and P uptake as well as the ratios C:N and C:P followed by measurements over 14 days in the experiment was retraced by the model.Task 2.6 Prototype Development and TestingBased on the findings from Tasks 2.1 – 2.5 prototype microalgae production systems will be installed and operated at the premises of SME AG members - for flue gas remediation at an AEBIOM member site; for aqueous effluent remediation at a BTA member site. The performance of these prototypes will be assessed and tuned in relation to model parameters (Task 2.5); the prototypes will also be used for Demonstration purposes in WP 7. Based on the findings in the upstream work packages a prototype PhotoBio Reactor (PBR) was designed and built at SU. The reactor was constructed in a greenhouse to give access to natural light. The system is a module design that gives greater flexibility with regards to volume, flow rate and layout.A bespoke monitoring and control system was designed, assembled and installed on the reactor by TI staff. Following a period of commissioning, testing and optimisation the reactor performance was assessed under a range of operating conditions. To be able to support the flue gas remediation trials a biomass burner was installed in a heat proof cabin and a system of pipes, compressors and solenoid valves was installed to deliver the flue gases from the burner flue pipes directly into the PBR. The control system monitors pH within the system and controls the addition of flue gas accordingly. Further monitoring equipment was fitted to the flue gas supply line and the PBR off gas line to allow for monitoring of quality and quantity of these gases.The PBR was also an integral part of the demonstration and training work packages WP7 + WP8. The successful installation of microalgae production technology at a partner site with delivery of a trout farm effluent based nutrient source and flue gases from burning biomass materials.At the outset of the project it had been intended that prototype microalgae production systems would be installed and operated at premise of the SME-AG members. However, at project meetings and through discussions with the BTA representative and his members it became clear that the process of operating, maintaining, finding a secure and suitable environment for the systems, preparing and analysing the reformulated nutrients and subsequent collection, handling and preparation of samples for analysis was not going to be practical at a member site. It was agreed by all partners that deploying the PBR at SU would be much more suitable as analytical equipment and facilities and suitable trained staff would be on hand to conduct the trials. It was also for this reason that the flue gas trials were also transferred to SU combined with the seasonal operation of the NOBio biomass plant that had previously been identified as the preferred site for deployment of the PBR. This meant that this particular biomass plant in Norway only ran during the winter months which was the least optimal time of year for growing algae if using natural light. So with the full backing of all project partners the resources planned for deploying a reactor in Norway were instead used to provide flue gases to the reactor at SU.Work package 3 – Microalgae Harvesting TechnologiesWP 3 – Objectives• To develop methods for harvesting the selected microalgae species with specific reference to costs and operability;• To develop primary separation strategies based upon a sound physical basis using properties of the organisms and the process fluids;• To provide a basis for choosing the harvesting process for different microalgal products;• To investigate batch and continuous process operation (including in situ recovery).In reporting period 3, activity and progress include the following tasks:• Task 3.2: Optimisation of Conventional Harvesting Technologies in the Context of Microalgae Culturing Systems.• Task 3.3 Characterisation of Microalgae Concentrates and Their Preservation• Task 3.4 Investigation of Hybrid Harvesting Technologies: Electro-FlocculationWorkPackage 3 is successfully completed.Task 3.2: Optimisation of Conventional Harvesting Technologies in the Context of Microalgae Culturing Systems.Operating envelops will be assessed for different harvesting methods assessed in relation to the physical properties process fluids and microalgae cells, and cultivation systems This data will allow calculation of capital and operating costs of the operations. The rheological and gross composition of concentrated materials and their drying characteristics will be determined.Cross-flow microfiltration of the selected microalgae species will be carried at pilot scale to establish filtration characteristics during concentration; centrifugation methods will be tested at pilot scale using a disc-stack system; sieving methods (drum filtration) will be tested using laboratory scale apparatus involving a series of stainless steel and plastic meshes in the range 25 micron 100micron. The data obtained for each harvesting method will allow for optimal operating conditions, design and scale-up, together with a comparison of capital and operating costs at full scale operation.The harvesting of the four microalgae species using a pilot-scale microfiltration system was investigated and critically assessed. Draft report on Task 3.2 was uploaded to Basecamp on the 3rd August 2012.Scenedesmus species was used as a species model and its filtration energy requirement was determined and correlated to the different operating parameters such as transmembrane pressure (ΔP), membrane area, temperature and initial biomass concentration. Addendum to task 3.2 was uploaded onto Basecamp on the Monday 5th August 2013.’Filtration parameters for all four species were determined with the respective filtration model. Using Scenedesmus species as a model species, the energy requirement and carbon footprint of the process was determined.Task 3.3 Characterisation of Microalgae Concentrates and Their PreservationThis task will characterise the physical and chemical composition of concentrated microalgae biomass and develop processes for stabilising these concentrates in preparation for further downstream processing / valorisation. 4 Different preservation techniques will be compared, namely freezing, drying, freeze-drying and spray-drying. Gross biochemical composition and product quality (highly unsaturated fatty acid composition; protein functionality) will be compared for the different methods, including assessment of shelf life.The total lipid content and total carbohydrate content of the 4 selected microalgal strains has been investigated after they have been preserved using freeze drying, spray drying, oven drying, refrigeration and conventional freezing for 1 day and 43 days. The pigment and protein content of the algae after the various preservation techniques was completed. Findings from this task will be compiled into Deliverable 6.4.There was a significant reduction in the total lipid content of refrigerated microalgal cells after 43 days of refrigeration (P<0.05 T=2.94 St Dev= 1.53 DoF=5). There was a significant reduction in the total carbohydrate content of refrigerated Scenedesmus samples after 43 days of preservation (P<0.05 T=3.78 St Dev, 2.12 DoF=5). There was no significant difference in the protein and pigment content between the different preservation techniques.Task reported in deliverable 6.4 as it was felt that this data fitted more appropriately with the remit of that deliverable (financial considerations). Completion of this task was delayed from RP2 into RP3, but was still completed in time to feed into downstream work packages.Task 3.4 Investigation of Hybrid Harvesting Technologies: Electro-FlocculationThis task will investigate the utility of electro-flocculation for harvesting the selected microalgae species, to establish the most appropriate operating conditions, and the limitations and potential of this harvesting approach. Based on laboratory-scale experiments, estimates will be made of harvesting costs as a function of microalgae concentration and comparisons made with traditional biomass recovery processes (section 3.2).It was agreed within the project partnership that the work described in this Task is sub-optimal for harvesting microalgae for value added applications. Instead, it was agreed to investigate a hybrid approach using microfiltration-centrifugation (see notes from BioAlgaeSorb Management Meeting, Brussels Mon 25 and 26/6/12), a pre-concentration membrane filtration step followed by centrifugation as a final concentration step, for the production of algal pastes.Investigation has been completed into the sedimentation rates of the different algal species under varying centrifugation rates. This work details the sedimentation properties under a centrifugal force for each algae species. This allowed for the determination of the hydrodynamic behaviour and the centrifugal parameters of each algal species. Centrifugation technologies seemed redundant due to their high energy consumption and also inadequacy for the established downstream processing of microalgae. The research group had instead focused on the obvious merits of membrane filtration and thus further developed methodologies for minimising the energy expenditure and associated carbon foorprint. This work was uploaded on Basecamp on the 20th on November 2012. See the appendix for further details on the costs of centrifugation. See Addendum to Task 3.2. for further details on minimising the energy expenditure and associated carbon footprint of microalgae harvesting.Work package 4 – Refinery and Upgrading Processes for Microalgae BiomassWP 4 – Objectives: To fractionate and convert harvested microalgae biomass to obtain high value products, utilising all the materials to obtain chemicals, energy and recycled / cleaned process water. The focus will be on the production of valorised stable materials to serve as feedstock and ingredients, e.g. highly unsaturated fatty acids (HUFAs), proteins, carbohydrates and minerals (silicates), leaving the remaining materials available for thermal transformation (WP5) and/or reuse via anaerobic fermentation and digestion and combustion of methane.In reporting period 3, activity and progress include the following tasks:• Task 4.1: Disruption of Microalgae. • Task 4.2 Physical Separation / Fractionation Schemes for Fractionation of Disruptates to Produce Stable Materials• Task 4.3 Chemical Conversionso Task 4.3.1 Heterogeneous catalysiso Task 4.3.2 Enzymatic reactionso Task 4.3.3 Fractionation of phycobiliproteinsWorkPackage 4 is successfully completed including Deliverable D4.1 (resubmission – draft submitted in RP2) and Deliverable D4.2.Task 4.1: Disruption of Microalgae. The harvested materials obtained in WP 3 will be used as raw materials for upgrading. Physical cell disruption has been shown to allow far better extraction of valuable materials than by direct solvent extraction or pressing. This task will investigate the best conditions of microalgae cell disruption (homogenization versus sonication) and will include investigation of the possible pre-treatments and optimisation of environmental conditions to minimise power consumption and ease further downstream processing. The disruption characteristics of the four selected algae species were established using a high-pressure homogeniser. Across the range 0-40 kPSI the disruption efficiency was assessed by monitoring the release of metabolites into solution. It was noted that each species had an inherent optimal disruption pressure and that up to 80 % cell disruption was possible in some cases. The disrupted biomass was amenable to further downstream processing (Task 4.2). A non-deliverable joint report on Task 4.1 and 4.2 was submitted on the 25th of February 2013. Each algae species had an optimal disruption pressure and in some cases up to 80 % cell disruption was possible.Task 4.2 Physical Separation / Fractionation Schemes for Fractionation of Disruptates to Produce Stable MaterialsThe microalgae disruptates from Task 4.1 will be treated using a combination of MF/UF and NF filtration systems to fractionate, wash and desalt the materials. In addition, specific absorption processes will be used to separate and refine protein fractions based on charge and hydrophobicity. Finally, the fractions obtained will be stabilised by freezing and/or drying. Four primary fractions will be produced: oils, soluble proteins, insoluble proteins and carbohydrates; further fractions may also be obtained depending on origin of the disruptates, eg silica from diatoms. The quality of these fractions will determine their value and as such the processing must be rapid and efficient. Chemical composition of the stabilised fractions will be measured and specifications for the processes determined.The work here reported focus on the downstream processing (DSP) of microalgae concentrates. The fractionation of soluble materials was carried out using membrane filtration technology. We have investigate (1) cell disruption using a high-pressure homogeniser, (2) physical separation of soluble materials using microfiltration technology especially targeted at proteins, and (3) fractionation of the soluble materials exploiting the differences in molecular weight by using ultrafiltration technology. Fractions rich in proteins and/or carbohydrates were obtained and lyophilised for further chemical characterisation. All four microalgae species have been processed in this fashion.A joint non-deliverable report for task 4.1 and 4.2 was submitted on the 25th of February 2013. The adopted fractionation strategy allowed to recover multiple fractions from microalgae disruptates: protein rich and carbohydrate-rich fractions and finally lipids extracted from the biomass debris.Task 4.3.1 Heterogeneous catalysisThis task investigated three chemical conversion processes (trans-esterification, decarboxylation and pyrolysis) employing heterogeneous catalysts. (1) Trans-esterification reactions for conventional biodiesel production were trialled, using super-basic layered double hydroxide materials. (2) Ketonic decarboxylation of fatty acids for ketone production was carried out using layered double hydroxides and novel mixed metal oxide catalysts in batch systems. (3) In the pyrolysis process, zeolites and mixed metal oxide catalysts were investigated under batch pyrolysis conditions to see if the high molecular weight oils can be cracked (broken down) to provide lower carbon number compounds for fuel applications. Significant results were achieved for each process, which are detailed below:*Trans-esterification reactions: Potassium tert-butoxide exchanged layered double hydroxides with Mg/Al molar ratio from 2 to 5 were prepared and used as catalysts for trans-esterification reactions of ethyl hexanoate, with the presence of methanol. The reactions were performed at 65 °C for 3 h. A high conversion of ethyl hexanoate was obtained, up to 80%, which indicated that tert-BuO-LDH catalysts were efficient for trans-esterification of ethyl hexanoate with methanol.*Ketonic decarboxylation reactions: Mg-Fe LDHs intercalated with CO32- (molar ratio of Mg/Fe = 2, 3, 4 and 5, respectively) were prepared by co-precipitation. Decanoic acid was used as the substrate for testing the catalytic performance of Mg-Fe LDHs. 10-nonadecanoane was the target product. The effects of reaction temperature, reaction pressure and catalyst loading were investigated. MMOs derived from calcination of the corresponding Mg-Al and Mg-Fe LDHs were used as the catalysts as well. About 90% of ketone yield was achieved under the optimised reaction condition, 300 °C for 3 h, with a very low Mg-Fe-2 MMO catalyst loading (mass ratio of decanoic acid/catalyst = 15).*Pyrolysis reactions: In order to explore microalgae as a potential feedstock for biofuel/bio-chemicals production, not only the whole N. oculata but also the lipid-extraction residues of N. oculata were used as the feedstock for pyrolysis reactions. ZSM-5, HZSM-5 (Si/Al molar ratio = 23, 50 and 80, respectively) and the basic material, Mg-Fe-2 MMO were used as catalysts. The pyrolysis results indicated that HZSM-5 (Si/Al molar ratio = 50) with moderate acidity is a potential catalyst for pyrolysis reactions of the both types of feedstock. A high bio-oil yield can be obtained with a relatively low content of acids. When the whole N. oculata was pyrolyzed with Mg-Fe-2 MMO, bio-oil with a lower content of acids was produced, but the bio-oil yield was also relatively low, compared with HZSM-5 catalysts. A higher bio-oil yield can be obtained from pyrolysis of the lipid extraction residues with Mg-Fe-2 MMO. However, the content of free acids increased, which makes the produced bio-oil more corrosive and less attractive for further applications.The pyrolysis reactor came online approximately 7 months behind schedule. The reasons for this delay were two-fold: i) the project started 6 months late at Durham owing to the recruitment process starting on project start date and the process, coupled to obtaining a visa for Dr Li taking just under 5 months; ii) the reactor required some dedicated space which took some time to clear, a delay which was compounded by ongoing issues with the reliability of one of the heater modules, which was subsequently traced to a wiring issue, beyond the project team’s immediate control. However, during the interim period, Dr Li was able to prepare a range of catalysts to trial, as well as completing a literature report and some of the work for later objectives. The draft report of D 4.1-Month 24 - “Report on pyrolysis of microalgae fraction using heterogeneous catalysts” was submitted on time. The final version of D 4.1 was submitted and uploaded onto the basecamp website as soon as possible, after all experiments were done.Task 4.3.2 Enzymatic reactionsUnder this task, trans-esterification reactions for conventional biodiesel production were trialled, using nanosponge (a polymer support material, obtained from Sea Marconi) supported enzyme catalysts. Trans-esterification reactions of glyceryl trioctanoate, similar to the main non-polar lipid components of microalgae oil, were performed with the prepared immobilized enzyme catalyst. The enzyme Pesudomonas cepacia was immobilized on cellulose polyurethane by adsorption and covalent binding. Ethanol was used for trans-esterification reactions, without any solvent. To figure out the optimal reaction conditions for the immobilized Pesudomonas cepacia catalyst, effects of temperature, water content and reaction time were investigated. Immobilized Pesudomonas cepacia showed both great thermal stability and water resist ability at 35 °C. 47.8% conversion of glyceryl trioctanoate was obtained at 35 °C, with 5.0% of water content, during a reaction period of 12 h.Task 4.3.2 was planned to be carried out during the last 6 months of this project. However, it was more difficult than expected. The task was about enzymatic catalysis and need a lot of knowledge and hand-on experience in biology, which was big challenge for Dr Li, who is a chemist by training. Dr Li developed all the needed characterisation and reactions, with considerable effort. Significant results were obtained and the deliverable D 4.2 was submitted with an allowed delay, without any further impact on this project.Task 4.3.3 Fractionation of phycobiliproteinsPhycobiliproteins are another high value component of microalgae (used as colouring agents, dyes, and marker compounds), which are present at very low concentrations. Previous research had shown that certain mineral matrices might be extremely effective at concentrating these compounds. In the proposed research, fractionation of phycobiliproteins was to be tested in temperature gradient flow reactor systems packed with various mineral substrates. However, a literature review of phycobilliproteins revealed that many of these structures are uncharged or of low mass to charge ratio, and therefore difficult to intercalate in layer minerals. Initial experimental work showed this approach to be unfeasible as a technique.Owing to this task not being included in neither D 4.1 or D 4.2 according to the project proposal, no corrective action was needed in terms of deliverables, however we have attached the literature review at the end of D 4.2 for completeness.Work package 5 – Thermo-chemical Conversions of Microalgae BiomassWP 5 Objectives - Pyrolysis has been the subject of massive investigation in the last decades by the scientific community, due to its promising efficiency and reliability for both production of chemicals and power. The consortium will benefit of this background because extensive know-how in pyrolysis reactor design has been gained. The specific objectives of WP 5 are:• To design and test a pyrolysis reactor to process microalgae• To study the feasibility of bio-oil production from four species of microalgae• To characterize the bio-oils with respect to fossil fuels• To identify critical issues for long-term performance analysisIn reporting period 3, activity and progress include the following tasks:• Task 5.3 Chemico-Physical Characterisation of Products• Task 5.4 Preliminary Tests for Heat and Power ProductionWorkPackage 5 is successfully completed including Deliverable D5.2.Task 5.3 Chemico-Physical Characterisation of ProductsThe oil product properties will be investigated, in order to identify the better strategy for its use in power and heat production. The analysis will be based on the existing standard and literature on the subject of pyrolysis oil characterization: the produced pyrolysis oil will therefore be analysed according to this standard. The 5 most critical parameters, in view of pyrolysis oil use in energy generation technologies, are: composition, viscosity, acidity, stability, carbon residue. The oil physical and chemical characteristics will also be evaluated versus transport fuels.The activities of this task were focused on the characterization of the chemical and physical properties of the bio-oils from microalgae, obtained by intermediate pyrolysis at 450°C in the CREAR semi-continuous pyrolysis reactor. Samples characterization included both analyses on the feed (microalgae) and on the pyrolysis products. Feeds were analysed in their elemental composition (Carbon, Nitrogen, Hydrogen, Sulphur, Phosphorus, Oxygen), compositional analysis (protein, carbohydrate, lipids), proximate analysis (ash, volatile, fixed carbon), thermo-gravimetric behaviour (heating-up in inert atmosphere at constant heating rate until 850-900°C). In addition to the above mentioned analyses, bio-oils were further analysed in their acidity, Conradson Carbon Number, viscosity change with temperature, corrosiveness toward metals (copper, aluminium, AISI 304 and AISI 316L) and elastomers (NBR, VITON, silicone) and other properties to provide a comprehensive and complete characterization of the material. Liquid products were analysed in their constituents with the aid of a GC-MS. For each oil, identified compounds were grouped in classes and their relative abundance compared among the three microalgae species. Within this task, since only few reference protocols could be found for the analysis of the organic phase of bio-oils from microalgae, new standard procedures for analysis were developed and applied. In this task, CREAR laboratory also supported project partner Durham University (Dr. Greenwell and Dr. Li Li) carrying out on their behalf the analysis on the micro samples of pyrolysis oil produced in their laboratory for their scheduled activities.A complete characterization of 6 samples of liquid products, i.e. bio-oils from three different microalgae strain, each originating organic- and water-rich fractions, was carried out along with a characterization of the intact feed. The amount of microalgae provided by partners was sufficient to obtain data for the characterization of bio-oils, but not enough to obtain sufficient amount of bio-oils for testing in the power generation test-rig, i.e. to power an engine or micro gas turbine. New methods for the analysis of pyrolysis products were developed.Task 5.4 Preliminary Tests for Heat and Power ProductionThe produced bio-oil, if a sufficient amount will be available, will be tested in a modified prime movers, i.e. a microturbine or a small diesel engine, in order to verify its characteristic as a substitute for fossil fuel replacement. This relatively straightforward test will be carried out in our facility, using standard mineral diesel oil as a control for comparison. Recommendations for long-term operation, if possible, will also be given.Due to the limited amount of microalgae biomass supplied by project partners, the quantity of oil that could be produced in the CREAR pyrolysis unit was in the order of 350g for each microalgae specie. These amount of oil where scarcely sufficient for laboratory analyses, and therefore, as already anticipated to partners since the very early project meetings, it was not possible to carry out real scale power generation test, e.g. feeding the oil to the CREAR micro gas turbine. Instead of testing the bio-oils in a real biofuel test bench based on a micro gas turbine or a engine, in this task the activities focused one evaluating the property of the bio-oils from microalge with respect to other fuels used in CHP application. It was chose to adopt a reference fuels a diesel and a kind of pyrolysis oil from pine chips. Results from the study were presented at partner in several progress and technical meetings.According to the analyses carried out in task 5.3 the suitability of bio-oils from microalgae was analysed with respect to the possibility of feeding an engine with such bio-oil. It was found that unless upgraded, e.g. through a hydro-treating process to decrease the O2 content and a further hydrocracking / hydro-decarboxylation, bio-oils form microalgae are not suitable for feeding and engine for CHP. Milestone M8 “first repeatable production of bio oil” was achieved, as showed in Deliverable 5.2.Work package 6 – System Integration and Industrial ValidationWP 6 – Objectives: Integration of the sub-processes produced in Work Packages 2, 3, 4 and 5 in order to obtain a fully functional microalgae-based effluent treatment system. Validation of the BioAlgaeSorb technology by demonstration of the fully integrated technology against the Objectives.In reporting period 3, activity and progress include the following tasks:• Task 6.1 System Integration• Task 6.2 Industrial Validation / Benchmarking• Task 6.3 Modelling Approaches for Economic Evaluations• Task 6.4 Risk Assessment and Contingency ManagementWorkPackage 6 is successfully completed including Deliverable D6.1 Deliverable D6.3 and Deliverable D6.4.Task 6.1 System IntegrationBased on the findings from task 2.1 – 2.5 a prototype microalgae productions system was designed and built at the premises of Swansea University. At the outset of the project it had been intended that prototype microalgae production systems would be installed and operated at premise of the SME-AG members. However, at project meetings and through discussions with the BTA representative and his members it became clear that the process of operating, maintaining, finding a secure and suitable environment for the systems, preparing and analysing the reformulated nutrients and subsequent collection, handling and preparation of samples for analysis was not going to be practical at a member site. It was agreed by all partners that deploying the PBR at SU would be much more suitable as analytical equipment and facilities and suitable trained staff would be on hand to conduct the trials. It was also for this reason that the flue gas trials were also transferred to SU combined with the seasonal operation of the NOBio biomass plant that had previously been identified as the preferred site for deployment of the PBR. This meant that this particular biomass plant in Norway only ran during the winter months which was the least optimal time of year for growing algae if using natural light. So with the full backing of all project partners the resources planned for deploying a reactor in Norway were instead used to provide flue gases to the reactor at SU.Prior to the design and construction of any new prototype photo bioreactor (PBR), a survey of existing technology currently employed for the pilot to large scale cultivation of algae was made in order to compare and contrast the relative advantages and disadvantages of the available options and to identify which, if any, reactor type best suits the needs for the current application.The prototype of the photobioreactor built at Swansea University includes the following elements:*A module design that gives greater flexibility with regards to volume, flow rate and layout. *A bespoke monitoring and control system was designed, assembled and installed on the reactor by TI staff. Following a period of commissioning, testing and optimisation the reactor performance was assessed under a range of operating conditions. Further information is provided in the Deliverable D6.1 Report.Task 6.2 Industrial Validation / BenchmarkingAlgae-to-bioenergy technologies are still pre-commercial and are in need for significant R&D to increase productivity, reduce costs and need to find other product market combination that gives a synergetic effect.AlgaeBioSorb will combine several technologies boosting the financial viability and will make this form of alternative energy successful and financially viable. The project has performed and industrial validation and benchmarking of biodiesel sources and production costs. Chemically, most biodiesel consists of alkyl (usually methyl) esters instead of the alkanes and aromatic hydrocarbons of petroleum derived diesel. In Europe, commercial use of biodiesel began after the year 1980. Its usage has greatly increased, in particular during the last few years. Biodiesel production in the European Union (EU) has increased approximately more than 2.5 times in the last 3 years rising from 1.9 million tonnes in 2004 to 4.9 million tonnes in 2006 . The directive (2003/30), promoting the use of biofuels in transport, issued by the European Commission in 2003 is important for this increase. The Directive created two indicative targets for EU member states: 2% biofuels penetration by December 2005 and 5.75% by December 2010 . In the EU, 82% of the total biofuel production is biodiesel3. The amounts show that the EU is the worldwide leader in both biodiesel production and capacity. In the year of 2006, the EU produced about 77% of the biodiesel produced all over the world. The USA is the second largest biodiesel producer in the world.The industrial validation and benchmarking include description of biodiesel production from different raw materials including vegetable oils, spent/used oils as well as micro-algae. Biodiesel production methods is outlined as well as biodiesel economy including algal biodiesel, soybean biodiesel and biodiesel from spent/used oils. Further, future European Policy within this sector is investigated. Biodiesel has become an attractive diesel fuel substitute due to its environmental benefits, the main problem is still the high costs surrounding biodiesel production and must be solved in order for it to be competitive in the fuel market either as a blend or as a neat fuel. The largest part of the production cost is associated with the feedstock itself and consequently, efforts are focused on developing technologies capable of using lower-cost feedstock, such as recycled cooking oils and wastes from animal or vegetable oil processing operations. Microalgae are also an interesting possibility due to the extremely high yield of oil per unit area.The findings and activity in Task 6.2 is specified in a separate task report.Task 6.3 Modelling Approaches for Economic EvaluationsThe product stream, based on nutrient-rich waste waters with an certain expense for disposal, used a feed for the growth of microalgae is examined. To appreciate the whole process of algal production requires the coupling of a description of the process itself with that of an economic model that takes account the economics of running the new processes. The task bases on the modelling approaches for technical evaluations (Task 2.5).In the current model for technical evaluations a module called ‘commercial production index’ was integrated to upgrade the existing technical model (Task 2.5) to an economic model. The module allows to balance the needs to obtain high volumic production and high areal production which is a requirement to optimise processes in open pond and closed photo bioreactor, respectively.The model was run to simulate annual Biomass, Lipid and Carbohydrate production and its costs at different nutrient levels and dilution rates (chemostat operation) in two different photobioreactors (PBRs) like they were used in the project: 1. the 600 L horizontal tube Varicon PBR at SU; 2. the 1800 L vertical tube PBR at HCMR. For both BBRs two locations were assumed to demonstrate the model’s latitude module: 1. Crete; 2. Oslo. The total production costs were composed of the costs for the growth medium, for N and P as fertilisers, lighting, harvest (dewatering the cultures to a slurry) and drying. The consequences for the whole process when some of the costs can be reduced (e.g. replacing P fertiliser by P rich waste water from trout farms) where demonstrated.The model simulations show, simple optimization of operations has potential to impact significantly on commercial viability; operating PBRs in an “intelligent” fashion has great potential for enhancing not only profitability, but also providing a tuneable crop, optimised for biomass, fatty acid, and production of other valuable products.For an economically successful production the cost reduction of electricity is crucial (e.g. using own wind turbine on the site). Then feeding a nutrient waste stream from a trout or dairy cattle farm into microalgae production might be an option. The possible savings by using flue gas as CO2 source appear to be fairly minor.Task 6.4 Risk Assessment and Contingency ManagementThe risk assessment and contingency planning include monitoring of progress in the different RTD WorkPackages in order to monitor and evaluate the progress of both the WP and its inter-relationship with the entire project. Inter-relationships between individual and particularly high-risk work packages and tasks have been evaluated. In order to monitor and minimize risk, good communication between RTD partners has been highly focused. Technical meetings are held in association with the Management Board meetings. The main objective of the Technical meetings have been to discuss the project results and RTD activity in further detail and especially plan and coordinate the RTD activity for the next 3-6 months period. Separate agenda and minutes are prepared for the Technical meetings. In addition to the Management Board meetings and the associated Technical meetings listed, a very high number of less formal technical meetings have been arranged to discuss the technical aspects of the project. These meetings do included physical meetings between a limited no. of beneficiaries as well as skype and telephone meetings.Potential Impact:The potential impact of the BioAlgaeSorb project include 3 different sectors, i.e. waste water treatment, biofuels production and greenhouse gas emissions: • Waste Water Treatment - in parallel to measures to decrease European GHG emissions, the discharge of aqueous effluents is increasingly regulated across diverse business sectors within the EU. Eutrophication is caused by several industrial activities (Tusseau-Vuillemin (2001) and it has long been suggested that water pollution should become an international priority (Duda, 1993). There are some 20 EU Directives encompassing all aspects of water quality, including those affecting water abstraction and effluent discharge from agriculture and land-based aquaculture, municipal waste water treatment and food and drink production. Key legislation affecting businesses that produce soluble organic wastes include the Water Framework Directive (2000/60/E), the Urban Waste Water Treatment Directive (91/272/EEC, amended by Commission Directive 98/15/EC); The Nitrates Directive (91/676/EEC) and the Integrated Pollution Prevention and Control Directive (2008/1/EC). These regulations are driving both better conservation of water (e.g. water re-use and recycling in land-based aquaculture systems) and upgrading of effluent treatment infrastructures, all of which have significant cost implications for SMEs and large enterprises in Europe. As an example, the EU Court of Auditors estimate the Europe-wide cost of constructing new sewer pipelines and secondary treatment plants in compliance with the Urban Waste Water Treatment Directive to be about € 200 billion (The information centre, Scottish Parliament 1999). There is clearly a need to develop innovative, cost effective approaches to the treatment of effluents, preferably incorporating valorisation of wastes. Critically, as the global cost of fertilizers increases (N is fixed by the Haber process at great energetic cost, while natural P reserves are fast being exhausted) it will become all the more important to recover and reuse these nutrients rather than to deal with them as wastes. The EC Water Framework Directive and related legislation place great pressures upon the removal of this valuable resource; it is logical to combined the removal of nutrients with the production of biofuels. The BioAlgaeSorb project will provide EU SME groups in the livestock production sectors with new technologies implementing microalgae for effluent treatment – phycoremediation – yielding a valuable by-product in the form of microalgae biomass.• Biofuels Production - The EC is committed to a target of 20% energy production from renewable sources by 2020. As part of this scheme, biofuels are to comprise 10% of European transport fuels by 2020, however biofuels have recently been the subject of much criticism within the EU and globally, for diverting human food supplies and arable land to fuel production (euobserver.com). Photosynthetic microalgae can be cultured to produce biofuels that do not directly compete with food crop-based commodities, as they are not typically grown in arable land areas, nor is microalgae biomass a major food source for humans (although it is a high quality food supplement). Furthermore, microalgae naturally tend to produce a lipid fraction suitable for the manufacture of second generation transport biofuels that are compatible with current transport infrastructure and do not require vehicle modification. Also, microalgae produce higher oil yields (up to 50 % of algal body weight) than oil-palm trees (up to 20 % body weight) which are currently the largest producer of oil to make biofuels (Research and Markets). It is therefore very timely to develop technologies for microalgae mass cultivation in Europe as a source of environmentally sustainable, carbon neutral biofuels.• Greenhouse Gas (GHG) Emissions - For the past two decades, reduction of GHGs has been high on the political agendas for the European Union, individual Member States and worldwide organisations such as the United Nations. Recognizing that anthropogenic activities contribute significantly to climate change, the EU has adopted ambitious targets for reducing GHG emissions in the coming decades. This has led to current and emerging GHG mitigation agreements and incentives such as: The Kyoto Protocol, The EU2020, ref Directive 2003/87/EC, Greenhouse Gas Emissions Trading and national agreements such as the UK Carbon Reduction Commitment. The current target of 20% reduction in EU GHG emissions by 2020 will not be achieved without significant reduction of CO2 emissions from power production, where the use of fossil fuels, primarily coal and gas, leads to approximately 40% of all CO2 emissions EU-wide, and totalled almost 5,000 million metric tonnes in 2005 (Europa report; International energy annual, 2006). These decreases are expected to be attained by reducing the carbon footprint of existing fossil fuel-based power generation and by developing alternatives to fossil fuels. Geological carbon capture and storage (CCS) from fossil fuel-based power plants is a current focus for technology development in the EU (e.g. proposed EC Directive on Geological Storage of CO2) and globally, however geological CCS will not be adaptable to all scales of operation or localities (e.g. where seismic events and other geological failures may cause broken pipes): additional technologies are needed to reduce GHG emissions from fossil fuel-based power plants. The BioAlgaeSorb project will benefit SMEs and other enterprises by developing technologies for biological carbon capture using microalgae.List of Websites:The BioAlgaeSorb website (www.bioalgaesorb.com) was established “up and running” within 3 months of the project start. It will be a useful means of communicating non-sensitive information about the project to the wider public and provides a useful portal for members of the BioAlgaeSorb consortium to login to the password protected project site. The project web site has been updated with new information representing progress and results. The front page of the project web site does present the overall project description, all beneficiaries of the project and contact details. Through the presentation of the different project beneficiaries - links are made to their respective home page. The front page also includes a "Search" function and provides facilities to contact the project representatives. The project web page has a seven “page” set-up; • Home• Project Descriptions• Project results• Beneficiaries• Contact• Related informationWhen entering the web-page, you are shown the “Home” page and can move around to the other pages as needed, either by clicking on the tabs displayed in the top part of the page or by selecting the links on the pages. The FP7 logo and the EU flag logo are both visible on the upper, right side of each page and it is cleared stated that the “Project is funded by EU Seventh Framework Programme” at the bottom left of every website page. Under the 2 logos, it is possible for all members of the BioAlgaeSorb consortium to login to a password protected website, where all information, included sensitive information about the project is stored. This is to ensure good and full communication about the project within the consortium.English is the main language of the project and reporting and dissemination is primarily done in English.