On this page we present three key problems we solved thanks to the immense knowledge and expertise of the stakeholders we talked to. In these problems, we synthesise how we integrated the information we gathered from multiple stakeholders in order to solve those problems, and the main conclusions we reached after all the brainwork we did. We have focused our narrative in answering the main questions we had throughout the project and that fell into the different 6 W’s umbrellas. We have also included one way of closing the loop with AlgaGenix, taking into account the needs and concerns of a local community affected by water scarcity.
We also present the timeline of meetings and reflections we had with all the stakeholders we contacted throughout the AlgaGenix journey. In order to make it more clear for the reader, we curated all the information we got from the actors we got in touch with and we explained how we integrated their feedback into our project. Additionally, every box contains the type of stakeholder (colour and hue) and its scope of action (badge), so the reader can easily locate the specific actor in the stakeholder map.
Problem 1: main focus
Summary
We decided to tackle nitrate pollution because of the magnitude of the issue and the impact it had on our society. We decided to address this problem through the enhancement of the nitrateabsorption pathways of Chlamydomonas reinhardtii, a microalgae model organism with which our PI and the laboratory we were going to do all the wet-lab in were familiarised. However, we acknowledged the existence of other better-suited microalgae, but for now we would focus on the model organism C. reinhardtii.
The first big setback we faced at the start of our project, as surely happens with the vast majority of iGEM teams, was answering two basic questions: “Which local issue do we want to tackle?,” and “How can we use synthetic biology to solve that problem?”
After we constituted our team and during the first team meetings and brainstorming sessions, we realised most of us were from different corners of Catalonia. Our region is divided into four provinces, and we somehow managed to put together a team with representatives from three of them. Thus, when deciding the topic of our project and the local problem we wanted to tackle, we looked for something that affected all of us, a problem that most of the team could relate to and understand, something that felt close to home.
As we explain in the main article of our magazine (and the “Why nitrate pollution?” section in the “Our approach” page), we decided to tackle nitrate-polluted water. We soon realised that nearly 3 million people were potentially affected by this issue, and in fact, some of our team members lived in nitrate-vulnerable zones and were experiencing the water restrictions first-hand.
Once we selected the problem we wanted to address, however, new questions arose: “How do affected communities deal with the issue?” “To what extent does the problem have a toll on the everyday lives of affected communities?” “When did this problem start?”
To answer the first two questions, we contacted Marc Magem, a Biotechnology student and classmate of some team members. Marc lives in Torelló, a town from Osona, one of the regions with the largest and most productive pig farms in Catalonia. As a result of this, they couldn’t drink water from underground wells and water reservoirs, because they were heavily polluted with nitrates. The World Health Organisation sets the maximum allowed nitrate concentration in drinking water at 50 mg/L. However, in towns like Torelló, the nitrate concentration exceeded that threshold by even a nine-fold. Marc told us that they had to take their water from the Ter, a river that flows through northern Catalonia and is free of nitrates, and that locals knew that drinking from the local fountains posed a threat to their health.
To answer the last question, however, we talked with Antonio and Beatriz Pérez-Portabella, father and daughter engineers with strong ties with the farming industry. They explained to us that during the 1990s, a huge surge in pig meat production happened in Catalonia, and intensive farming activities magnified to unprecedented levels for the region. Due to a lack of effective legislation, this led to the rapid pollution of underground water reservoirs. Nowadays, 40 % of Catalonia’s surface is considered a Nitrate-Vulnerable Zone.
Of course, we asked ourselves: “How can we help them?” “How can we use synthetic biology to fight against nitrate pollution?” At first, we met several times to brainstorm possible ideas on nitrate-absorption, but we didn’t know which organism and through which methods could we achieve that.
That is when we met our future PI, Jae-Seong Yang, a researcher from CRAG (Centre for Research in Agrigenomics) who worked with microalgae, specifically Chlamydomonas reinhardtii. He offered us his laboratory to conduct all our wet-lab activities, so we started considering working with microalgae as well. We did some research and found that microalgae could do photosynthesis, so they could produce their own biomass, and also they usually could survive in higher nitrate concentrations. In addition to this, we studied the nitrate-absorption pathways of C. reinhardtii and rapidly understood the ways we could enhance them to increase that activity.
However, we found out that C. reinhardtii wasn’t the optimal microalgae to tackle this issue, but since we were just identifying the solution and this was a well-established model organism, we thought that the most convenient approach was starting with Chlamydomonas and, if everything was successful, extrapolating all our knowledge to other better-suited microalgae. Later on in the project, we got to talk with different experts on microalgae use (Aigars Lavrinovičs and Manuel González del Valle) in bioreactors and they all proposed other microalgae with which our process would be more efficient.
Note: to fully understand how we chose this topic and how we understood its full magnitude and scope, we recommend reading the main article in the magazine we have created. In it, you will find a more developed and holistic narration combining the experiences of many stakeholders and the feelings we had throughout the project.
Problem 2: bioproduction
Summary
After painstaking research and many meetings with experts and stakeholders, we concluded that the format of our desired final product would be a cytokinin-rich microalgae lysate in liquid or spray form, which in turn would also be rich in other fertilizing substances such as phosphorus, nitrogen, sulphur, etc. In order to measure cytokinin production in C. reinhardtii, we decided to focus on 3 specific ones: tZ, IPA and ZR, for which reliable detection measures were accessible and well-established.
Once we established that we wanted to enhance nitrate uptake with microalgae, a pressing issue we tackled was around the question “Can absorbed nitrates be transformed into a valuable compound?” Our PI, Jae-Seong Yang, was categorical: yes. Our initial question, then, was transformed into “What can we produce with the absorbed nitrates?”
At first, we considered different basic molecules whose biosynthesis pathways were relatively close to the main metabolic pathways of all microorganisms: amino acids. This way, we avoided having to enhance an excessive number of genes to ensure a straightforward implementation of nitrogen to the desired molecules. However, after talking with Dr. Cristina Madrid, we realised that most anabolic pathways were heavily regulated at different levels, especially in relation to essential biomolecules like the ones we were contemplating. We then thought about producing proteins –maybe this way we could produce a high-value compound with another application outside the nitrate-affected sector. Despite that, Manuel González del Valle, CEO of G2G Algae Solutions (a company that uses microalgae in many different projects and applications), told us that there were other alternatives way more efficient than microalgae to synthesise proteins, mainly bacteria.
After a few brainstorming sessions, our PI proposed synthesising biostimulants, molecules that stimulate natural plant processes. We started researching immediately which ones could we produce with Chlamydomonas reinhardtii, our model organism, and we came up with three different options: gibberelins, auxins and cytokinins. The first two, however, didn’t contain any nitrogen in their structure (or just one atom per molecule), so we had to discard them. On the other hand, cytokinins (CK) did have some nitrogen atoms, so we focused on this last group of phytohormones.
At this stage, we had the following questions: “What is a cytokinin?” “How do cytokinins act as plant biostimulants?” “Does C. reinhardtii naturally synthetise these compounds?” This time, Google had all our answers: CKs are adenine-derived molecules (therefore with at least 5 N atoms per molecule) that act as biostimulants in plants at incredibly low concentrations, usually regulating stress-response mechanisms and gene expression. Also, most plants and algae naturally synthesise cytokinins at low concentrations, and our Chlamys (at this point we had already nicknamed our organism) were no exception. We were all set, then; we wanted to transform the absorbed nitrogen into cytokinins.
We started researching the pathways of CK biosynthesis, and we found that there were two main genes implicated in the process: LOG and IPT. Taking into account what Dr. Madrid had warned us about, we delved into the regulation systems of both genes, and we found that there was regulation only at the transcriptional level, that is, we just needed to use a strong promoter to effectively enhance the desired activity of the genes.
Once we had both the nitrate-uptake and CK-production pathways and strategies well established, we just needed to close the gap between them. We realised that a crucial enzyme was implicated in that interaction: Glutamine Synthase (GS). This enzyme is the perfect example of regulation at all levels of the gene-to-activity process in E. coli. Since Chlamy’s GS regulation systems are not well characterised, we decided to bypass all the possible regulations by introducing an exogenous GS from an unrelated species that required two specific proteins to be fully deactivated –two proteins that our Chlamys did not naturally synthesise. This way we ensured the constant expression and activity of the key mediating enzyme of our process.
Now that we had all the pathways established, the next set of questions started to take shape in our minds: “How are we going to measure CK in C. reinhardtii?” “How can we isolate the produced CK?” “How are CKs applied to plants?” At first, we were unable to find a clear and straightforward answer to all these questions, so we decided to contact Dr. Sergi MunnéBosch, a renowned UB professor whose main area of research conveniently revolved around cytokinin production and measurements.
He told us that CK-detection protocols for trans-zeatin (tZ), isopentenyladenine (IPA) and ribosidezeatin (ZR) were well established and that he could process some measurements for us, so we decided to focus on those specific cytokinins. However, we had doubts regarding the isolation of these molecules at an industrial scale, and he told us that it would be easier to lyse the cells and apply the mixture directly to the field.
Problem 3: application
Summary
After consulting with many stakeholders, we concluded that for now we would focus on absorbing polluting nitrogen from underground waters in a batch or fed-batch photobioreactor, from which we would obtain clean water and a biomass concentrate rich in cytokinins, which could be lysed and even turned into powder to be sold as fertilizer. This process should be in the form of a pilot water treatment plant. We would need an effective way of ensuring the elimination of all microalgae like UV radiation in a system similar to that of the CAT water-treatment plant. This way we would be one step closer to adapting to GMO regulations. Lastly, in the future, we could implement our project directly on farms without nearby arable land, but first, we should ensure the correct functioning in a preliminary pilot plant.
Throughout the project, we faced several questions around the application of AlgaGenix: “Where can we apply our project?” “Who can benefit the most from AlgaGenix?” “Who is being left out of the solution we are providing?” We rapidly understood that these were trickier questions that required consulting many actors, experts and stakeholders to properly answer them.
To tackle the first question, we contacted Manuel González del Valle again, and he told us that due to the nature of our microalgae, we should contemplate a closed and properly-sealed bioreactor because ours was a Genetically Modified Organism (GMO). He also proposed that an interesting “place” we could use AlgaGenix was in space, since up there there are no GMO regulations and the recirculation and optimisation of resources are of utmost importance to the field. Later on in the project we also contacted Aigars Lavrinovičs, the mind behind Algae Phos4x (a Latvian startup that aims to absorb polluting phosphorus from water using microalgae), and he also believed that our project could be of great potential in space investigation.
Even though this application seemed fancy to us, we felt that it wouldn’t allow us to focus on the people we had wanted to help in the first place: the local communities spread throughout Catalonia that couldn’t rely on underground water reservoirs to source their water. Taking this into account, we decided to talk with a major local stakeholder: bonÀrea, one of the leading meatdriven supermarkets in northern Spain and the flagship of Corporació Guissona, a company with over 4,500 associated farmers. Ramón Gabriel, Director of Agricultural Contracts from Guissona, told us that only farmers that don’t have lands to dispose of the slurry would be interested in AlgaGenix, thus limiting the application of our project in the “ground zero”, that is, right at the origin of the problem.
Taking this into account, we decided to talk with two other local stakeholders: the Cooperativa d’Artesa de Segre and the Comunitat de Regants Sindicat Agrícola de l’Ebre. These entities regulated the activity and water usage of thousands of farmers in Lleida and Tarragona (the easternmost provinces of Catalonia) and agreed on what Ramón Gabriel had said: that only farms with no available land to throw their manure in would be the ones that would be interested in our project.
To get a different point of view, we got in contact with the Agència Catalana de l’Aigua (ACA), the public company in charge of water planning and management of Catalonia, to understand the intricacies and main concerns of the regulatory body of the water sources of the region. They told us that nitrate pollution was as worrying as the ongoing drought and that Catalonia was in pressing need of new and more efficient ways of treating manure. That is why they told us that if our Chlamys worked, they would let us try them in a pilot plant in Osona, the epicentre of nitrate pollution in underground waters. This fact, in addition to what bonÀrea and the other local stakeholders had told us, made us decide on a “water-treatment plant” approach, that is, instead of directly implementing AlgaGenix on farms, we would first aim at a more controlled and manageable application like a small water-treatment plant, and from there we could properly see our next moves.
In addition to this, G2G Algae proposed a collaboration to use one of their photobioreactors specially optimised for Chlamydomonas reinhardtii, so it was clear that AlgaGenix, even at its earliest stages, was seen as a potential game-changer in the field. He also told us different tips to increase the efficiency of our product at an industrial level: we should use sand filters, the only ones that work with Chlamys, to separate the water from the biomass, and then we should lyophilise the latter in order to obtain an easy-to-store and easy-to-use powder.
However, we still had one last question to answer: “How can we address GMO regulations?” We visited the water treatment plant of the Consorci d’Aigües de Tarragona, where we realised how everything in a water treatment plant was tightly regulated and controlled. They explained to us that just the previous year they had included an additional step based on UV radiation to kill microorganisms. That’s when we realised we could use something like that in order to efficiently deactivate our modified microalgae, in the first step of the lysing process. This way, we would be ensuring no recombinant DNA was left in the water, an essential requirement for GMO-related services.
Closing the loop
As a part of our university studies, some of our team members were taking a course on Biochemistry and Plant Physiology where the laboratory part was focused around the use of a biostimulant in Solanum lycopersicum, a tomato variety. We saw an immense opportunity in this, since we could be able to test the effect of CK in this plant.
We thought about the different ways of using kinetin, a type of CK, to expand AlgaGenix, and after some literature reading, we found that they were associated with salinity-response stress mechanisms, so we decided to follow this path. Our ultimate goal was being able to use moderately-salty water (as a result, for instance, of the mixture of fresh water and sea water) to irrigate crops in areas of high water stress. This idea developed from our visit to the Comunitat de Regants Sindicat Agrícola de l’Ebre, whose main issue was high salinity because sea water infiltrates into the soil of the delta and increases the salinity of the soil.
Since we were applying the project at the local level, the Mediterranean Sea has a saline concentration of 38 g/L, so we used this as a reference of “salty water”. After some research, we found that 100 mM of NaCl was considered as a “severe stress” for S. lycopersicum, and the concentration of the sea water of reference was of 651 mM. This is why we decided to dilute the sea water with fresh water, and focusing on how finding a good concentration with which our CKs could mitigate the effects of high salinity.
This was, therefore, a clear application of our final product in a way that further contributed to the recirculation of our resources and that directly tackled the concerns of a local community severely affected by freshwater scarcity.
Timeline
14/3/23 - Reflection (STARTER)
Assessment
As this is our first meeting, the most important goal was to get to know the team and start brainstorming possibilities. We learnt the necessary information about the competition to move the project forward.
Analysis
We did not brainstorm this session but decided to trace the general theme in which we wanted to work this year, bioremediation and agricultural improvements, as to be able to do individual research from home.
Conclusion
We still do not have a set project, but are starting to gather ideas to move it forward.
Action plan
Each of us has to gather information for the next meeting and present a project proposal to pick apart and brainstorm.
15/3/23 - Jae-Seong Yang
Aim
General state of our team, referring to the meeting we had the previous day and showing the lab facilities to the team leaders.
Contribution
He thoroughly explained to us what they were performing in his lab in order to have more ideas for our project that could relate with his research. They were working with microalgae, so maybe a project using modified chlamydomonas could be a good idea because they knew how to perform experiments with this microalgae and had the facilities for that.
Key highlights
They worked mainly with microalgae and they were giving us all their facilities to help us with the project.
Action plan
We had to think about project ideas involving microalgae mainly.
28/3/23 - Reflection
Assessment
The goal of this meeting is to brainstorm about the ideas we have developed individually, and present the different rough projects we have developed for the moment, as to study viability. We also need to prepare our first team-collaboration and team picture.
Analysis
Five different projects are put forward: mycorrhizae for droughts and denitrification, tobacco mosaic virus, nitrate water contamination, use of biocharcoal, and protein production in algae. We decided to further work on them to test how viable they are by finding literature and writing drafts to present to the team.
Our main focus has been drawn to the nitrate crisis in Catalonia, and how to solve it by working in agricultural settings.
Conclusion
Further research in the project topics is needed to solidify the strongest contenders for the project.
Action plan
We have divided the team to work on each of the five proposals, so we can assess their function, impact and viability during the next meeting.
8/4/23 - Reflection
Assessment
The team needs to reassess the different project proposals and find weak points in the different projects to be able to improve them and choose the best fit for our team.
Analysis
We have dismissed two of the projects, mycorrhizae and biocharchoal, and currently evaluate whether combining nitrate contamination and protein biosynthesis in algae is possible. Currently, Our main concern is how to tackle agricultural nitrate treatment, whether to directly treat the animal excrements, or to treat the contaminated water that derives from them.
Conclusion
Further research needs to be conducted to rule out projects or study whether they can be combined into one. We have to find solutions to the problems proposed by each of the projects and evaluate whether we can overcome them.
Action plan
We need to conduct more individual research to finalise this initial brainstorming / project drafting step. We also need to draft official project proposals to send to our PIs to get feedback and constructive criticism from them.
12/4/23 - Cristina Madrid (University of Barcelona)
Aim
Dr. Cristina Madrid is a UB professor with both a degree and a doctorate in Biological Sciences. She specialises in molecular microbiology and her area of work revolves around stress response in bacteria. We wanted to consult with her the endogenous regulation systems of various compounds in order to understand the level of difficulty producing each compound had.
Contribution
She told us that Glutamine Synthetase (GS), one of the enzymes we were planning to enhance, was heavily regulated in practically all microorganisms, so we had to find a way to bypass that regulation.
She showed us different bioreactors and photobioreactors from which we could start ideating the bioreactor we wanted to make as part of one of the ideas for the project.
Key highlights
We have to be careful with endogenous regulation systems for all the enzymes we want to enhance/reduce.
Action plan
We have to do more research on what molecules we want to modify and how they are regulated in the model organism we will be using. We also have to find a way to bypass said regulations.
18/4/23 - Jae-Seong Yang and Ivan Reyna
Aim
Discuss with our PI and another one from CRAG about the three project ideas we had.
Contribution
They gave us their opinion about the project ideas and thought that the nitrates uptake idea with chlamydomonas was feasible. They also liked the bioproduction idea, and said that, even though it would be more difficult, it would be more attractive to the general public.
Key highlights
The best option was to modify Chlamydomonas reinhardtii and make them absorb or produce something. Both ideas together were a really good starting point.
Action plan
We had to continue our bibliography research and start planning the project design taking into consideration that the best ideas were using microalgae to absorb nitrogen and produce something.
Assessment
Due to the need of finally grounding ourselves with only one project, we are going to decide which project proposal to move forward with. As we cannot delay his decision further, our goal is to finally decide on a project so we can focus all our team’s energy on it.
Analysis
We did a final set of brainstorming to get on the same page then reviewed the project proposal together and sent it to our PIs. We have decided to unite nitrate water contamination and biosynthesis in algae into a cohesive project based on the circular economy principle.
At last, the mosaic virus project was rejected due to the safety concerns involved around working with the virus.
Conclusion
We now have one project divided into two main blocks: nitrate contamination bioremediation and algal biosynthesis, that need to be linked together. We are waiting to hear back from our PIs to incorporate their feedback into our project.
Action plan
We are researching the necessary data to make the union function better, and have given individual research tasks to our members in order to solve found problems and find pathways and bioproducts compatible with our project proposal.
11/5/23 - Reflection
Assessment
We had many things to tackle in this meeting as we needed to discuss the project name, review the feedback from our PIs, find genes for the project, and choose which bioproduct best fits our concept.
Analysis
The feedback from our PIs is positive towards the project proposal we drafted last meeting, and they suggest potential bioproducts for our algae. This is currently our biggest incognita as there are many benefitting products that could be of interest, each of them with different biosynthesis pathways.
– There is one main product we are considering, auxins, but we need to evaluate its feasibility.
– After internal discussion, we decided to reject this proposal, as literature suggest they are not compatible with high nitrogen-level mediums.
– Currently, we are weighing our options, which are to either modify the algae to make it more compatible with our interest or to exchange our bioproduct to a simpler one, such as aminoacids.
Two different methods for nitrate absorption were evaluated, but the choice relies on which final product we will synthesise and its metabolic pathway.
– We have encountered some inhibitors in nitrate transporters, but have laid out a plan to override the internal block.
We’ve discussed potential structures of the bioreactor to better suit our project, and we are concerned about extraction methods that can potentially separate our bioproduct from the modified algae.
Conclusion
For now, we have decided to move forward with aminoacid production as our bioproduct, therefore discarding the other proposals. On the other hand, we still need to find ways to override nitrate transporters inhibition. However, the main incognita for our project right now still remains: deciding which bioproduct we can make from our algae, and how to extract it.
Action plan
We need to conduct a Bioinformatics study for protein/amino acid folding to see how the planned changes on them might change their functionality and/or structure. We have also set out to find out what regulates nitrate canals. Genes involved in nitrate transporters need to be researched so as to finish adding all the genes and pathways on our project proposal. We have to conduct individual research about the currently considered bioproducts as well as methods of extractions of bioproducts from inside the algae.
2/6/23 - Manuel González del Valle (G2G Algae)
Aim
He is the CEO of G2G Algae, a company that uses microalgae to clean waters, so we thought that it was a great idea to talk with an expert who had a business related to our project idea. We wanted to show our idea in order to validate it and get a different point of view of our project.
Contribution
The project is promising, he liked it. However, he said that there are other pollutants as important as nitrates: phosphates. He recommended that we explore this area.
About production he didn't tell us a specific product, but to explore the metabolic pathways and find an intermediate product. He said that producing proteins is a good idea, but he wouldn’t produce them as there are easier and more effective ways to produce them.
Chlamydomonas is a good model, but it is a mobile microalgae. Maybe we should change the model. Not during the experimental part, because chlamydomonas is more used in laboratories, but when presenting the project, we should state that there are better microalgal models.
Explore the idea of bringing our project to space. It could be a very promising idea as there are no regulations on transgenics and water treatment is a must.
Key highlights
Find a better product to produce, as well as try to combine nitrates and phosphates and bring our project to space.
Action plan
We will study the phosphates assimilation route to see if it is feasible to be done. We will explore better the metabolic pathways of microalgae to find a better product to produce. We have to study the idea of bringing our project to space.
27/6/23 - Jae-Seong Yang and Ivan Reyna
Aim
This meeting aimed to establish the basic guidelines of the laboratory work and to define what needed to be done in the lab before starting with the actual experimentation/wet lab. Additionally, this was an opportunity to solve some doubts we had regarding microalgae regulatory metabolism and some technical aspects of working with Chlamydomonas reinhardtii.
Contribution
They have told us to focus on producing cytokinins, a different fitohormone with many N atoms in its structure.
The idea of using PCR amplification to get the endogenous gene’s sequences instead of ordering them has been put in the table, this way we would avoid having to domesticate the sequences and purchasing them.
We will add a constitutive promoter to our genes to avoid transcriptional regulation by ammonia, and there are high chances there would be no need to do knockdowns of their endogenous versions.
First things that need to be done once we go to the lab are the cloning simulation of all of our constructs. Plus, checking for restriction enzyme recognition sites.
Developing a detailed diagram of the implicated genes in the nitrogen absorbance and cytokine production including promoters and regulators.
Key highlights
We have changed the bioproduct and are therefore researching and piecing together the needed information to use in the lab once we start. We're considering using PCR amplification instead of ordering endogenous gene sequences. We'll also add a constitutive promoter to prevent ammonia-related transcriptional regulation, and we’ll create a detailed diagram of the genes involved in nitrogen absorption and cytokine production, along with their promoters and regulators.
Action plan
Further complete the diagram of the pathway including different promoters (in case the use of the same promoter causes undesired recombination of genes) and checking if the genes need to have import sequences to specific organelles.
1/7/23 - Concepción Novillo (Science Crop – Bayer)
Aim
Bayer is a leading company in the fields of healthcare and agriculture that focuses its research on searching solutions to improve the way farmers grow and protect their harvests, using land, water and energy in a more efficient way. We got in contact with Bayer to learn about their new ideas regarding the use and treatment of water and to get feedback for our project.
Contribution
She gave us an invaluable insight into the groundbreaking technologies and approaches the companies are planning on using in order to to assist farmers in making the most of all of their available resources, including their land and water. Moreover, she helped us understand the limitations of the European Union legislation, particularly the impact of the forthcoming new regulation on the use of GMOs in Europe. Additionally, She shared her thoughts on which stakeholders we ought to prioritise and who she thought our product could help the most.
Key highlights
Understanding the current situation of GMO laws and regulations in the European Union, and different approaches to help the agriculture sector exploit most of the resources at their disposal while implementing our product.
Action plan
Since GMO regulation legislation in the European Union is still quite restrictive, the idea of implementing our project in outer space has arisen. We should look more into the details of this, and how our product could be beneficial for cleaning water in spatial missions.
2/7/23 - Dorothy Zhang (iGEM)
Aim
At the ATG Synbio congress we got the opportunity to meet Dorothy Zhang, the vice president of Global Development at iGEM, we wanted to talk with her about our project, know her thoughts about our ideas and to get feedback on how to better make it fit within the context of the competition and it’s values.
Contribution
She attended our presentation during the congress and asked some really insightful questions that helped us figure out where to focus once the experimental results are in, such as the effectiveness of our system and identifying the target market for our product as well as its ultimate beneficiaries.
She gave us advice on how to better communicate the project, its values and potential.
We also got to chat with her during the networking breaks and share thoughts and opinions about the synthetic biology world and the values this community ought to uphold, such as integrity, responsibility, effort and respect.
Key highlights
Helped us notice weaknesses on our project’s presentation and helped us redirect attention to the most important parts of the project, in order to fit iGEM's values and criteria.
Action plan
Changing the narrative of our presentation so that we can focus and show the potential of AlgaGenix.
Where we are
The beginning of July came together with the International Synthetic Biology Congress held in Barcelona during the 1st and 2nd July and organised by ATGSynBio. We got the chance to participate actively in the organisation of some activities that took place during the congress (such as the iGEM International MeetUp).
It was also the first time we were participating in a scientific event of these characteristics as a team. Getting immersed into a scientific environment and presenting AlgaGenix with conviction and determination was a remarkable challenge. It was this way because we faced professionals from the field who could be of great help for further steps of the project but also had strong and reasonable opinions about it.
We have also started working at the laboratory at CRAG in July. With the help of Jae-Seong Yang we are finishing and checking our constructs, while we are learning about several culturing, measuring and genetic engineering techniques under the instruction of David Alzuria, Moritz Aschern and Angela Fermat.
Feelings
In the Congress, despite the initial uncertainty we felt when some of the ideas our project was based on were questioned, we got to overcome it. In consequence, we got to spot our flaws and fix them so they turned into new strong points. In addition, we got really good advice from several scientists and entrepreneurs that attended the congress.
On the other hand, about the lab, it is the first time that some of our iGEM team members are working on a laboratory, but it is everyone’s first time working with microalgae. Therefore, the first two weeks have to involve a lot of learning and habituation to the new laboratory environment. Once the techniques are mastered and the pieces have arrived, we will start the several cloning protocols that will lead to our more-efficient and transformed Chlamydomonas. We will definitely become more confident with our work and we will be getting our first experimental results.
26/7/23 - Sergi Munné-Bosch (UB)
Aim
Cytokinins are our main target as our phytohormones output-trade is based on the idea of improving the production of these substances. According to this point, we thought it could be useful to have an opinion from someone who currently works on these affairs and could help us with the detection, recognition and examination of our experiments and genetic transformations on Chlamydomonas.
Contribution
To provide an idea of how we have to carry the cytokinins detection (tZ, z-IP, ZR, IPA) as well as a pathwork for developing our experiments based on his validated method for HPLC/MS-MS (Müller, M., & Munné-Bosch, S. (2011)). This explanation gave us an idea of the magnitude the protocol has and a better vision of the samples managed in terms of amount, sampling through lyophilisation and storage (-80ºC).
Key highlights
Ways of detecting cytokinins, clarifications on the way of processing the samples and number of free samples he can analyse for us.
Action plan
To read the paper written by him and his colleague and to collect the dry weight needed for analysis.
28/7/23 - Antonio and Beatriz Pérez-Portabella
Aim
Antonio and Beatriz Pérez-Portabella are two engineers that have been involved in the pig industry for the past three decades, and have conducted different projects in order to adapt their farm to the new legislations and regulations that have arisen during this time. We wanted to contact them to understand how we got to this situation in the first place, and to know the different strategies that farmers use to dispose of the slurry their pigs produce.
Contribution
They explained to us the evolution of the pig industry in Catalonia and Aragon, starting in the 1990s, and gave us a holistic view of the legal and administrative situation of the problem we want to tackle.
They told us how to approach farmers when asking for delicate information like the one we want to ask.
They showed us the different possible ways a farmer can dispose of their slurry, and which techniques are in need of a renovation.
Key highlights
We got to the current situation due to a lack of legislation during the early 1990s, and even though now the government is very strict with the opening and keeping of pig farms, the slurry-disposal alternatives are unreliable and depend on market fluctuations, thus we are in need of a stable and definitive solution. Also, farmers might lie when approached about this issue because they do not want to compromise themselves or their neighbours, so a curated approach must be conducted.
Action plan
We have to set a strategy to contact the farmers and other rural stakeholders so they feel comfortable with us and see us as a motor for change.
30/7/23 - Marc Magem
Aim
Marc is a third-year Biotechnology student from the University of Barcelona. He is from Torelló, a town in Osona (province of Girona) surrounded by some of the biggest pig farms from Catalonia and in one of the nitrate-pollution hotspots of the region. We wanted to talk with him to understand how affected communities are coping with nitrate pollution and which sectors of their lives are the most affected ones.
Contribution
Marc explained to us that when his parents were young, they could drink from local fountains without fear, but that at some point during the late 1990s or the early 2000s, drinking from those fountains became a threat to the health of the population. He has never drank from a local fountain, and the nitrate issue has been a recurring topic in conversations between family and friends since way before he was born. He also stated that the ongoing drought has burdened even more local administrations to provide clean water to their citizens. A few decades ago every town had their own wells, but now they all drink water from the Ter, a river north from Osona that is free from nitrates.
Key highlights
He told us that now no one from his region wouldn’t even think about drinking from a local fountain in fear of nitrate intoxication, and that nowadays most towns depend on the water from distant rivers rather than local wells to source drinkable water.
Action plan
We will research how water is polluted in underground reservoirs and if there is a way for AlgaGenix to adapt to tackle that problem.
9/8/23 - Aigars Lavrinovičs (Algae Phos4x)
Aim
Algae Phos4x is a Latvian startup that aims to remove phosphates from polluted waters with microalgae. Instead of genetically modifying their organism, they are adapting it to different phosphate concentrations in a process called phosphate-starvation, so when it is exposed to phosphates again, it absorbs them with maximum efficiency. We contacted them because we wanted to know what working with a microalgae photobioreactor is like, and to see if he could help us redirect our project application since we doubted between water-treatment plants, the agricultural sector or hydroponics.
Contribution
The algae biomass can absorb pathogens and heavy metals: this fact cleans out the idea of how we can sell the green biomass or, at least, limitates the ways we could offer it (There’s legislation that doesn’t let you pour it or throw it as it remains after water-cleaning.) For him, they found that the best way to sell it was to freeze them. This could retain the main components and it doesn’t damage many others that are sensitive to heat-shocks.
We should try to focus on controlled facilities: at this point, we were thinking it could be applied to different water-cleaning facilities. Instead, the idea of using it in hydroponic cultures, vertical agriculture and horticulture, could be more useful and optimum as the waste-water remaining is not rich in pollutants. This would also promote the circularity of our project.
MELISSA pilot plant: according to what the expert said, there’s a pilot plant which is developed in our university where they are trying to find out new ways of processing the waste produced by astronauts and turn it into food, air and even water through microbiological pathways. We should ask more about it as it could also give us an idea of how we could present it and the protocol they are following.
Heterotrophic pathway of the Chlamys: the idea of developing a better heterotrophic-enhanced media where they can grow faster, is an excellent option for discarding a photoreactor as it doesn’t limitate the microalgae just to do photosynthesis; but help it to get an organic-carbon source easily.
Easy-to-use bioreactor: the hypothesis of creating an easy-to-use mechanism for farmers could facilitate an explanation to sell the idea and save some details that could not be understood by everyone. At least, it is the ideal scenario where you could just throw it in some water and let it grow. We shall consult the end-users about what would be the easiest way to use the product.
Key highlights
We should definitely focus on hydroponics/vertical agriculture or the space; we should look for using heterotrophic pathways as the growth system for our microalgae; and since the contents of the water we filter will directly influence in the market viability of our final product, so we should focus on environments free of pathogens and heavy metals.
Action plan
We will start contacting hydroponic companies to ask the viability of our project and how it can adapt to them. We will also contact space projects like MELISSA, but it is not our priority. How environmental factors such as heavy-metal content in polluted waters affect the final quality and viability of our biomass is something we hadn’t considered and we shall investigate further.
11/8/23 - Milestone
Where we are now
We have been working in the laboratory for more than a month. Although the initial struggles, it seems like the both parts in which we have divided our project (cloning and cytokinin/nitrate absorption) are working (a bit slowly though).
At the end of July, the team members who were at the laboratory left for holidays and they were substituted by others. The new members of the investigation team were assigned one of the projects and were taught by the most experienced team members so they could continue their work efficiently. The cloning team has assembled the level 1s successfully and the cytokinins/nitrate abortion team are quantifying nitrate absorption in different concentrations of nitrates in ammonia and no-ammonia conditions. They are also doing growth tests of different strains of algae in different supplemented mediums.
We have also established the final application of our project. We have started considering hydroponics as a really likely strategy of application of our final product. Therefore, we could design a bioreactor in which the nitrates would be absorbed by our modified Chlamydomonas, and the cytokinins produced would be used as a fertiliser in hydroponic agriculture (in the same bioreactor complex or nearby).
Feelings
The change of team members at the lab performing the experiments has been initially a bit messy and it has felt a bit like a setback. Nevertheless, they have soon managed to learn and master all the protocols and techniques.
It has also been satisfying defining a more specific application of our project and realising that it is actually viable (under the opinion of several professionals).
Despite that, we have started realising that soon August will be over and a lot of lab work must be done before that. It is slightly stressful and we will need to start getting along with the idea that not all the transformations will work the way we wanted them to.
17/8/23 - Manuel González del Valle (G2G Algae)
Aim
After speaking with the CEO of Algae Phos4x, we started looking for more specific and technical information about our wastewater treatment process. Especially regarding methods of biomass extraction and types of photobioreactors available. We also wanted to ask about the possibility of applying AlgaGenix to the hydroponics sector.
Contribution
He said that the cost of the biomass extraction process is high because of the process to remove the water, to separate the algae from the water. There are several methods, but the most commonly used are centrifugation and filtration. Centrifugation is better but more expensive. We will have to see if we can finance the energy costs.
There is a clear value to be added to make up for the costs of getting the biomass. This is where the cytokinins come into play and how we sell them.
About our application, pig manure is too concentrated. It is very difficult to work with it. To solve the problem of concentration, you could mix the pig slurry in the olive slurry pond, called “alpechín”. The “alpechín” is a dark, black liquid with a very unpleasant smell. It is a mixture of the water used to wash the olives and the water that the olives themselves contain. It is made up of 83% water, 15% organic matter and 1% minerals.
As for the application in hydroponics, it works very well indoors, but outdoors, in the open air, it does not do well. In hydroponics, there is a problem when you reuse the water because you can get plant diseases spread through viruses.
He likes the space application very much, and thinks that in the not too distant future it would make a lot of sense. We should contact Melissa immediately as they are very powerful and they have very similar projects.
Tubular bioreactor are the most expensive but the best, by far. They are also the most efficient. If we sell a very expensive product, the cost of the tubulars will be compensated. It will depend on our final product.
Key highlights
Does the cost of our final product outweigh the expenses of expensive but more efficient techniques? If we manage the type of photobioreactor and our filtration process.
Pig manure is very concentrated, but we could dilute it. Diluting in clean water will need a lot of space and water. If any farms with pigs also had olive trees, it could be diluted in the olive washing water. We should dig further into the space application.
Action plan
Inform ourselves about the price of cytokinins, and how much we could sell our product for. Also, we must say how much algae and water we would need to produce the cytokinins necessary to be a competent product with other biostimulants on the market? From there, see what processes would be profitable for us. We will contact Melissa.
28/8/23 - Marc Valls (UB)
Aim
Dr. Marc Valls is a UB professor with a doctorate in Biology who specialises in molecular genetics applied to agrobiotechnology. We decided to speak with him so he could review our project and help us with the application of AlgaGenix.
Contribution
At first we thought he was a researcher from IRTA, the Institut de Recerca i Tecnologia Agroalimentària, with whom we wanted to establish a collaboration, but it turned out he wasn’t from that institution. Either way, he was happy to review our project and gave us invaluable feedback on the presentation and how to approach both the young public and the international community. He also provided us with useful and interesting contacts with research advisory groups and institutions with whom we could contact and ask for an assessment of AlgaGenix’s viability and market application.
Key highlights
He made a thorough review of our project, gave us useful insight and provided us with contacts in entrepreneurship viability advisory bodies.
Action plan
We’ll make all the changes to the presentation he told us to do and we’ll contact the quality control and advisory bodies he spoke of.
30/8/23 - Joan Potrony (Cooperativa d’Artesa de Segre)
Aim
Cooperativa d’Artesa de Segre is a young catalan cooperative that strengthens agriculture and the livestock industry in some catalan regions, with a total of 1137 associates. As experts in agricultural fertilisation and production, we intended to get feedback on the viability of our product and cytokinin use in agriculture. Additionally, as they are investors in upcoming technology in the agricultural world, we hoped to assess whether our project would catch their interest or that of other sponsors.
Contribution
Nitrate pollution is of concern as legislation regulating their use and production are more strict by the year, to the point they have developed a special branch in the company to test technologies related to nitrate use.
Special interest was expressed toward the circular economy of our project, where we use a residue such as nitrate pollution to generate a product.
They recommended we provide further economic data and assess the scalability and continuity of our project to gather further attention and sponsors.
We must obtain data on the efficiency of our modified Chlamydomonas in both nitrate extraction from water but also cytokinin production.
If we intend to use the cytokinin as fertilisers in open fields, we must calculate the amount and methods. They advise us to use the cytokinin with excrements to fertilise fields as dissolving them in water is not enticing to the farmer.
Key highlights
They have offered us space to test our product once it is successful, and might invest once we have official results and data.
Action plan
The production of cytokinin has been the right direction to take to enrich our product, as it is the most interesting part of our product. We should consult and compare other fertilisers that use it, and highlight their importance in our pitches. It is very necessary to obtain yield and performance of our modified Chlamydomonas so we might produce the necessary data to solidify the project.
31/8/23 - Daniel Marsol and Ramon Gabriel (BonÀrea)
Aim
BonÀrea is one of the biggest emerging supermarkets in Spain, they are based in Catalonia and they specialise in meat production. This company puts together over 4.500 farmers from all over Spain, so they are a leading stakeholder of the meat-production market. We wanted to talk with them to make sure some of our data is correct, to further understand the slurry management in Catalonia and Aragon and to hear their feedback about a project like ours, as well as understanding the views at the other end of the pollution cycle.
Contribution
When asked about the implication of big supermarkets and cooperatives like bonÀrea regarding the slurry management of the farmers with whom they collaborate, they said that each farmer is responsible for the waste they produce.
Every livestock farmer is responsible for the waste they generate. Most farms are linked to a specific land base for the application of manure (the number of hectares is related to the farm's size), and each year they must report to the authorities how much manure they have generated and where they have applied it. Some farmers, however, don’t have enough land to dispose of all the slurry they generate. These farmers have to take the manure to an authorised treatment plant.
A small minority of farmers, on the other hand, treat the manure on their own farm using a method authorised and recognized by the authorities (solid-liquid separation, NDN, etc.).
They said our project would only attract those farmers that either have to take the manure to a treatment plant or have their own systems, and only if our alternative is more economical than the others.
They said the main problem our project has regarding a direct application with farmers is that they have to dispose of a watering system to manage the resulting water from our process, since transporting said water would be a waste of resources. The thing is that not everyone has a nearby piece of arable land to apply the cleaned water.
Key highlights
They evaluated our project and pointed out some insightful considerations about the application of AlgaGenix. Mainly, if we want to use our microalgae directly with farmers, only those that manage the manure through external treatment plants or other systems will be interested.
Action plan
We have to revise the application possibilities of our system with farmers, and explore other sectors that may be able to use the final water without having to transport it.
1/9/23 - Javier Casanova Casanova, Meritxell Jardí Llambrich (Comunitat de Regants Sindicat Agrícola de l’Ebre)
Aim
The Comunitat de Regants de l’Esquerra (Association of irrigators of the left side) are the main group of farmers in the Delta del Ebre. They are the ones who manage all the canals that bring water to all the rice fields in the delta. During the last years the water crisis has been their main concern as tons of water are needed to irrigate the rice fields and have a good harvest. Taking all this into account, we decided to talk with them to learn first-hand the impact of the water crisis and know better which are their main concerns.
Contribution
In the delta del ebro their biggest concern is the ongoing drought. About water pollutants, in the delta there isn’t much concern about chemicals or other products like nitrates. Their main worry is water salinity as they are next to the sea in an area with high values of salinity in waters and grounds. Moreover, they know about studies released before by IRTA about nitrates in the delta and will facilitate this information to us. Lastly, They are helping us spread a form to all the community of farmers so we can have data about how they treat their fields and know the impact of the fertilisers.
Key highlights
We should focus maybe in areas near Barcelona and Lleida where the nitrates pollution impact is higher. One concern they had about our project was the durability of the microalgae and also the genetic modifications, as the ebro delta is a highly protected natural space where there are strict regulations about new organisms.
Action plan
We will study the results of the form after we collect the responses in order to evaluate the amount of fertilisers they use and how they could be reverted with cytokinins. We will try to contact the contacts they gave us from IRTA. Lastly, we should study better the actual regulations of the transgenic organisms.
1/9/23 - Aleix Martorell (Consorci d’Aigües de Tarragona)
Aim
Consorci d’Aigües de Tarragona (CAT) is a nonprofit organisation with its own legal personality that collects, treats and distributes potable water to different towns and industries. Those towns are located in Tarragona, one of the provinces of Catalonia (Spain), and a huge part of the waters they deal with come from the Ebro river, one of the biggest rivers in Spain.
Contribution
We got to visit the water treatment plant guided by one of the workers there. We understood how water is collected into the treatment plant and through which treatments and different stages it goes. Finally, we visited the laboratories in which they test the water quality and they reassure that all the treatments are performed correctly and in an efficient way.
We realised how everything is tightly regulated and controlled: there are several control stations all along the water distribution path and water biological, organic and inorganic compounds are checked daily.
We could speculate where in all the water treatment processes our genetically-engineered Chlamydomonas could fit in and help improve water quality.
Key highlights
Any additional step that could be included into the water treatment process, has to be proven to be truly efficient and economically profitable. Despite that, changes are incorporated sometimes and new equipment could be acquired.
Nitrates are not the main problem of the lands that surround the Ebro river. They are more concerned about high salinity and sulfated pollution.
The main pollutants of the waters can change through the years: 10 years ago there was a huge mercury pollution, but nowadays this problem is not the biggest concern, since the levels have been lowered down.
Action plan
We will study these water treatment plants and others in Catalonia in detail to try to fit in our bioreactor. We should also focus on other terrains in which nitrate contamination is a bigger issue (Lleida and Barcelona) to try to sell our idea.
18/9/23 - Antoni Munné, Josep Fraile and Carolina Sola (Agència Catalana de l’Aigua)
Aim
The Catalan Water Agency (ACA) is the public company of the Generalitat de Catalunya that is in charge of water planning and management in accordance with the basic principles of the Water Framework Directive.
ACA promotes its action plan to guarantee, now and in the future, the supply, availability of water and its quality at source (groundwater and surface water). This action plan also promotes the sanitation of wastewater and the protection and conservation of water bodies and associated ecosystems.
Contribution
The main problem we have in Catalonia is the nitrate pollution of underground water bodies. Mainly due to diffuse agricultural pollution. Urban contamination is not as important. It is clear that manure is the one of the main contaminants, due to its extremely high nitrogen concentration. In the ACA, the method most used for manure management is biogas reactors.
Drought vs contamination? Does one outweigh the other? In Catalonia, right now, the balance is very even. The quality of the water is as important as the quantity. It is very closely linked because if you have little water, everything is concentrated, then the little water that remains will be even more polluted.
How does the ACA try to mitigate the contamination problem? The tasks of the ACA are mainly focused on the monitoring and control of the presence of nitrates in water, within the framework of which it carries out complementary measures to improve knowledge and the origin of nitrates. It sets specific restrictions to the industries that are pouring its residues into the water. They focus on monitoring and controlling, not on nitrate removal directly. It’s the owner of the farm or industry that is responsible for the treatment and the economic cost. Manure has always been considered a fertilizer. The problem comes when they are over-concentrated. Which is what is happening nowadays because of macro farms.
Do you know about communities or villages that are currently affected by nitrate pollution in water? There are many villages (listed on ACA’s webpage) that have had many difficulties to access potable water. In those villages, the agency has had to provide treatments to make that contaminated water drinkable. One of the main regions affected is Osona.
Faced with this context and with the desire to make a modern livestock sector compatible with the sustainable management of the territory from the Regional Council of Osona, the Table for the sustainable management of livestock in Osona was established. More popularly known as the "Table of manure" given the relevance of these in the environmental situation of the region. The general objectives of this Table are to pursue the environmental, but also the economic and social sustainability, of livestock in the Osona region.
They explain us about the Table and the possibility and interest in collaborating with them. Once we can prove to them that our algae are more efficient and cheaper than conventional methods, they are interested in setting a pilot test of our photobioreactor in one of their farms in Osona.
Do you see a future in AlgaGenix? Is it up to market needs? They assure that there is a clear need in the market of manure treatment. Nowadays, the methods used are expensive. They believe our project could be very useful if we can ensure a cheaper and more efficient treatment. If it were the case that it doesn’t turn out to be as cheap as they’d wish, they would still take into account the added value of the project, that is cytokinin production. We are generating a great product with very high interest that would possibly outweigh the extra cost of our water treatment.
Key highlights
Our application could be more focused on manure treatment, as there is a clear market need, specially in Catalonia. If we obtain competitive results in the laboratory, they would help us scale up and set a pilot test in one of their farms in Osona.
Action plan
We will start to focus on manure treatment. We will look into current techniques used and its cost. We will calculate theoretical efficiencies based on our lab results so far.
19/9/23 - Reflection
Assessment
Team reflection is meant to update everyone on the latest lab procedures, divide workload in the upcoming weeks and decide which prizes and medals we are most likely going to opt for.
Analysis
With the promotional video handed in, as well as the safety form, we are now pooling our main effort towards the wiki and result analysis. We divided the workload between our teammates according to whom best suited the task.
We discussed the final practical applications for our project, and have decided that it will ideally be sold as a tool for farmers or bigger industries with nitrate residues.
Other tasks to tackle now are the presentation video and further improving our education model, which we have also planned over the next weeks to accomplish.
Key highlights
We are most behind currently in wiki page editing and is currently our main priority.
As this is the last stretch of the project before the jamboree, projects have been assigned individually to the people best suited, and we hope to accelerate the process with this method.
We have decided to opt for the Human Practices prize.
Action plan
We are prioritizing the editing and writing of the wiki, overall, but other projects are also ongoing.
20/9/23 - Simon Maechling (Bayer Crop Science)
Aim
Simon Maechling is currently the Innovation Manager at Bayer Crop Science in Lyon, France. He plays a central role in identifying new innovation opportunities from Startups to established players to enrich the Research and Development (R&D) pipeline and technical platforms, particularly in the areas of Active Ingredient Discovery, Formulation Technology, Environmental Safety and Human Safety - Crop Protection of products in Agriculture. We were invited to a close event in Bayer’s LifeHub in Barcelona, where we had the chance to speak with the speakers.
Contribution
He really liked our idea, and the way we are transforming something of very poor value into high value products such as cytokinins.
He doesn’t think that water treatment plants are our ideal customer, because of the large amount of litres they are dealing with each day.
He also likes the idea about space due to the European regulations on the use of GMO for cleaning water for drinking. If not space, another market such as the American market, where the regulations are not too harsh.
He proposes that in the future if we scale up, we could produce cytokinins for pharmaceutical uses, as an active ingredient of body lotions or rejuvenating creams.
20/9/23 - Henna Niskakosk (CSO EOD Europe)
Aim
EOD ® Europe is a Finnish company that develops and manufactures innovative applications based on nanobubble technology. They have developed a Nanoboost device that increases the amount of oxygen in the irrigation water and makes the plants stronger. The amount of oxygen in the water is increased through nanobubbles. Adding oxygen to the water promotes root growth, improves plants’ stress tolerance and immune system, and enhances the absorption of nutrients.
Contribution
Oxygen nanobubbles in the irrigation water has already proven to be very beneficial for plants. They want to try their nanobubble-based technology on other organisms, like microalgae. We told her about our photobioreactor system, and thought it could be a good idea to try their technology with our algae. Instead of oxygen nanobubbles, it would be CO2-based nanobubbles. She sent us a few articles regarding how this would help growth, survival and nutrient absorption of the microalgae, including nitrate absorption.
Key highlights
CO2 nanobubbles could enhance the nitrate absorption and growth of our algae. We still don’t know about how the cytokinin production will be affected, but we believe that it will also promote its biosynthesis.
Action plan
We set a future date to meet and talk about a possible collaboration. They want to try their technology on algae. We think it would be a promising way to enhance the growth and efficiency of our microalgae in a photobioreactor system. We believe it will also help improve the algae’s survival rate.
2/10/2023 - Fidel Delgado (CEO neoalgae)
Aim
Neoalgae is a Spanish biotechnological company focused on algae applications for conventional industries: Live food of aquaculture, food additive, wastewater treatment, CO2 fixation and decreased CO2 footprint, pharmaceuticals and cosmetics and production of biofuels.
Contribution
He likes our idea very much. He explained to us what they were doing at Neoalgae; they are producing different products for different applications based on microalgae technology. They have different lines of work: cosmetics, nutrition and agriculture.
In the agricultural line, they produce biostimulants from spirulina extracts, which they sell in a liquid form. Liquid biomass is less messy and laborious to produce, but in order for it to be economically viable, the production should be at a large scale. No to mention the storage space necessary to store the products and the transportation costs.
Solid biomass doesn’t have these limitations, but you have to add extra steps in the processing of our biomass.
He also proposes that once we scale up, we could sell our algae biomass rich in cytokinins to companies that formulate bioestimulants or biofertilizers.
Key highlights
In the long-term, liquid biomass could work better than solid biomass. We could try and focus our project on bioestimulant production, and sell our liquid biomass to companies that produce biofertilizers.
Action plan
We could ask our mentors about the proposals of Neoalgae. Nevertheless, these proposals only seem possible once we scale up. Right now, we should focus on our initial plan.
6/10/2023 - Manuel González del Valle (G2G Algae)
Aim
After receiving a market analysis by bonÀrea, we realized that the project still had a number of uncertainties that needed to be resolved. Therefore, we decided to contact G2G Algae again, and get their opinion on the feasibility of our proposals. They have been our ideal mentors, as they have very similar technology and we have very similar applications.
Contribution
We take a look at our biomass production process. We continue the filtration vs. centrifugation debate to collect the algae. On the one hand, centrifugation is better, but more expensive because of the high energy cost. On the other hand, however, filtration is not as efficient, but it is cheaper. He says that conventional filtration will not work for chlamydomonas, as they are mobile and with conventional filters, chlamydomonas escape filtration.
We have to use sand filters in a differential pore. The problem with sand filters is that they are a lot of work and for some things they are very cumbersome. But he thinks it can work. Sand filters work with silica grains of different sizes. When you pour the water through the sand, the sand retains the biomass because of its electrical charge, and then the biomass is recovered by backwashing. We also have to lyse the algae, as they are GMOs. Lysis should be done when the algae are semi-liquid.
Biomass in liquid or powder form? Liquid biostimulant is usually easier to handle, but it is mainly water. We should come out with a powdered product, which allows the farmers to create different doses of the product, giving them the option of diluting the biomass to different concentrations, not a fixed concentration. All the companies on the market are already making biostimulants in liquid form, in the form of carafes. We must have a differential product to be able to differentiate ourselves in the market. He says farmers want to control their production a bit more and not have to use pre-set concentrations. We have to give them that freedom. Not to mention, transportation and storage. By having our final product in a solid phase, in plastic bags, we are saving a ton of space. If we were producing liquid biostimualnts, like the other companies, we would need a lot of space to store the bottles and a large truck to transport and deliver them.
We discussed the application of the project on pig manure. Are there any companies that are getting closer to solving the problem of pig manure? The problem is that it is too concentrated (concentrations of up to 8000 mg/L of nitrogen, according to a report by the Catalan Water Agency), and becomes toxic to algae. They have to dilute the pig slurry to 10% to be able to work with it, which is not economically viable. Large farms can produce up to 2 million litres of pig slurry per year. On the other hand, with cow and chicken manure you can work very well.
Hydroponics. In hydroponics, our cytokinins can do very well. The problem with hydroponics is not water with nitrates, it is microorganisms. By recirculating the water, it attracts, above all, the presence of viruses, which can damage crops and our algae. Right now what they are doing is recirculating the water from one plant to a different plant so that the virus from one does not affect the other. They are trying to treat with UV, but it usually does not work, since they work with thousands of litres and the UV does not penetrate to the bottom, or the water passes so quickly that it does not have time to eliminate the microorganisms. Viruses are very difficult to eliminate, and only if one survives do they replicate.
He believes that our project makes more sense to apply it in conventional agriculture, not in hydroponics. We have to sell the project by saying that we are transforming a waste product into a product with an application.
Key highlights
Right now, we can’t focus our project on biostimulant production. It is only economically viable at an industrial-scale. We should produce a solid final product, a biomass in the form of powder. We should filtrate our algae rather than do centrifugation. Conventional filtration won’t work with Chlamydomonas, we should try sand filters with differential pores. We should lyophilize our algae in order to dehydrate them and preserve the cytokinins. Better to stick to conventional agriculture, not hydroponics. Our project would make sense right now in the conventional agriculture market, as there is also a market need. In the hydroponics market, it is not the case right now.
Action plan
We ought to inform about the whole lyophilization process and sand filtration, and its cost. We will stick to conventional agriculture and selling a solid biomass product. Furthermore, we will need to verify if our algae can survive the high concentrations of nitrate found in pig manure. If not, we will focus on chicken and cow manure.
9/10/2023 - Sergi Compte (Catalan Water Partnership)
Aim
The Catalan Water Partnership (CWP) is a Catalan water cluster, and it is made up of companies and knowledge centres that have been operating in the sector of sustainable water use since 2008 and whose mission is to improve competitiveness overall of its associates.
Contribution
CWP is a cluster made up of more than 50 companies working in the sector of sustainable use of water and the cleaning up of water.
In Gerona, they have had many incidents with water contaminations due to nitrates, both in ground and surface water. They have a similar project with Chlorella, where they also use it to remove the pollutants from water.
They also work with many farms and farmers, and they would be very keen to working with us once we implement our project and establish as a startup.
If we consolidate as a business, we could enter the CWP association and have access to their stakeholers, advisors and partners, where we could have numerous possible collaborations.
Key highlights
Gerona is also significantly affected by nitrates. They have to resort to other water sources in order to drink and use water.
They would be very interested in collaborating once we establish our project.