Although the team recognized the poor ecological condition of the river and lake by researching the local problem of eutrophication in the literature and by walking along the river, this was not adequate. We were wondering, how the poor condition of these aquatic ecosystems impacts local people? What are their thoughts on the current state of the local environment, and do they have any idea what the underlying cause of the issue is? All these considerations inspired the team to contact local citizens of Thessaly and obtain feedback from those directly impacted by the phenomenon to determine their views, what they expect from society to do and what can we do to provide a solution to this emerging environmental problem.

As a result, we walked along the Pineios River to engage with local people and get their perspectives on the phenomenon and its impact on their daily lives. The river is often referred to by locals as the "lung" of the city; but, when we asked them what comes to mind when they think of the river nowadays, an awful view and odor comes to mind. One resident remarked, "Every year the situation gets worse, and now, we cannot even enjoy our everyday walk near the river”. Another concerned citizen mentioned “It is clear that the authorities have neglected the situation for years and there hasn’t been any new initiative in order to diminish it”. A large number of people acknowledge that they enjoy seeing and photographing the rich biodiversity of the river (fish, birds, etc.) It was also mentioned that due to the situation in the Pineios river activities such as fishing, and irrigation are not safe anymore.

Since then, we've discovered that a substantial portion of the local community realizes there's an environmental crisis but don't know the cause. They themselves have noticed the consequences of the problem, which is a decrease in the biodiversity of the river, as well as a lack of exploitation for economic support of the area through activities like fishing and tourism. Consequently, our mission as a research team was to both inform the public about the phenomenon of eutrophication and develop a synthetic biology-based solution.

Fisherman and Diver/Researcher, Curator at the Institute of Environmental Science and Technology of the Autonomous University of Barcelona

In an effort to learn about the problem of eutrophication in other regions of Greece from people who are directly affected by it, we reached out to Mr. Barbarousis, who provides us with informative insights regarding the ecological status of coastal bodies in Greece. In addition to his research on environmental issues, he is an active scuba diver in the coastal waters of Greece, particularly those around the islands of Samothrace and Limnos. He mentioned that he has observed green and brown mucus at the surface of the sea. This observation triggered him and inspired him to further analyze the phenomenon. More specifically, he took samples and measured temperature, pH levels, chlorophyll levels as well as the dissolved oxygen levels of the sea. These indicators' increased levels confirmed the eutrophication problem. Realizing the significance of the information, he went to report it to the proper authorities, but was unsuccessful. Therefore, he recommended that we should contact the local authorities in an effort to diminish the phenomenon. After our discussion, we understood the extent of the phenomenon in our country, and based on his recommendations, we contacted the local authorities to gather precise information and alert them about the problem.

Head of Directorate, General Directorate of Environment - Department of Water Economics and Supervision, Thessaly Region

To investigate the current situation in our region, it was essential to contact the local authorities of Thessaly responsible for water management, in order to receive information from them and alert them. Mr. Papamixail, in his capacity as a management officer, provided us with confirmation that Lake Karla and Pineios river suffer from eutrophication by underlining the significant environmental and economic effects on the Thessaly region. Managing these freshwater bodies properly is not a priority for the government, he claimed, therefore they are being devastated. Since these water sources are inadequate for agricultural irrigation, farmers are forced to resort to expensive and energy-intensive methods of obtaining irrigation water from underground aquifers, including pumping. He stated that the management of water bodies requires an approach that is efficient, environmentally friendly, and cost-effective in order to make the water clean and usable while preserving the local ecosystem. To this goal, we set out to develop an approach that might be effectively applied to these freshwater environments.

Meeting the Local Authority

President of the Lake Karla Management Body Field of expertise: eco-hydrology, management, and restoration of aquatic systems

To further understand the impact this phenomenon is having on the lake's biodiversity, we reached out to Dr. Ifigeneia Kagkalou, president of the Lake Karla Management Body. Firstly, Dr Kagkalou confirmed the eutrophicated status of the lake despite its enlistment in Natura 200 protected areas. Specifically, Lake Karla is home to 13 fish species and 180 bird species, some of them being extremely rare. She specifically mentioned the fish species Carassius Gimbelio and Gobitis Thessalicus as two of the most endangered species due to eutrophication. She also referred to the significant lack of monitoring data regarding the water situation in Greece. In an effort to cross-reference this information, we searched the web for open access data on the eutrophication status of various water bodies in our country but were unable to get adequate data. We hadn't considered a data gathering system before, but her advice triggered the idea of developing a water monitoring system that will provide real-time data and store it in a cloud where it will be accessible to the entire scientific community.

LIFE Project Coordinator at Hellenic Ornithological Society

Because one of the main impacts of eutrophication is the endangerment of species, and after hearing about the increased mortality rates of silver pelicans in Lake Karla a few years ago, we wanted to learn more about local bird species threatened by eutrophication. Therefore, we contacted the Hellenic Ornithological Society to determine the magnitude of the phenomenon's influence on the region's birdlife and biodiversity in general. Initially, when we asked Mr. Kordopatis about the issue of eutrophication, he appeared to be familiar with it and claimed that the study of a lake's ecosystem is quite complicated. As part of the One Health approach we investigate, we found it important to ask about how the health of the whole ecosystem can be affected by the phenomenon. He specifically mentioned that aquatic wildlife such as fish, usually die because of the low oxygen levels that might occur during the phenomenon and the toxins released to the waterbody. He highlighted that aquatic species, such as fish and amphibians, are more at risk from eutrophication, but diving bird species, such as pelicans, are also very vulnerable to the phenomenon. Regarding the incident that occurred a few years ago, they do not have enough evidence to conclude that it was caused by eutrophication, although it is quite likely that it was. Therefore, according to the Hellenic Ornithological Society's representative, the ecosystem's biodiversity is threatened, and a solution must be found to preserve it.

Meeting with Ornithological Society

President of the European Association of Environmental and Natural Resource Economists (EAERE)

Dr Kountouri, as an environmental economist, provided us with important information regarding the main consequences of eutrophication in both global and local economies. More specifically, she mentioned that eutrophicated bodies in Greece are proven to contribute to financial loss according to several sources that she also referred us to. The phenomenon can cause up to 2,2 billion US dollars global economic loss while affecting human, animal and environmental well-being. She also played a vital role in informing us about the SDGs that correlate with our project. Prior to that, we aimed to target different SDGs goals, and thus she assisted us in redefining our specific goals. Lastly, she advised us that to successfully implement our project we need to develop a cost-effective and sustainable approach for eutrophication.

Meeting with Dr. Kountouri

CCB Working Area Leader on Eutrophication and founder of CCB River University.

We couldn't possibly fully understand the extent of the phenomenon of eutrophication without examining the situation outside of our borders. Dr. Les provided us with essential information on eutrophication in the Baltic region and Europe. She first confirmed that agriculture is the primary source of the problem. More specifically, she stated that over 50% of nutrient inputs in the Baltic area originate from agriculture. As far as the impacts of the phenomenon, she mentioned the financial burden created. More specifically, algal scums increase the costs of water treatment to avoid the taste, odor, and cyanotoxin problems in the treated water. The expenses of cleaning and repairing equipment are also increased when large blooms block filters. After substantial eutrophication has occurred, the expenses of remediation might be enormous. Macrophytes may need to be sprayed or controlled using other biological or expensive interventions. On the basis of this information, our first thought was that we needed to eliminate the phytoplankton from the aquatic environment by focusing our efforts on the algae. The initial plan was to genetically modify bacteria to chemotactically follow a specific chemical and also bind to microalgae, so that when the chemical was sprayed to the banks, it would both chemotactically attract the bacteria and algae to the banks. Therefore, we may collect algae there using filters, rather than filtering the entire body of water using equipment.

Professor at the University of Thessaly, Department of Ichthyology & Aquatic Environment.

We contacted Mr. Kormas, who has extensive experience with eutrophication and contaminated waters, to seek his opinion on whether the idea of chemotactic attraction of algae on bank sides, might be used in fresh waters. But initially, he provided us with valuable knowledge regarding the eutrophication phenomenon. Due to his expertise, he was necessary in advising us that eutrophication occurs in areas with slow water flow or permanent water, such as lakes, stagnate river areas and reservoirs. We should prioritise lake ecosystems since lakes are more susceptible to eutrophication, as he warned. Before that, our team had intended to focus primarily on the riverine ecosystem of Pineios, but he supported his opinion by saying that lakes, and especially Lake Karla, are well studied ecosystems and have a handful of scientific data, in comparison to rivers that are a lot harder to study and evaluate the extent of eutrophication. Lake Karla is an artificial, shallow, hypertrophic lake and therefore provides more consistent and easier to measure variables. Furthermore, he explained that freshwater cyanobacteria species, specifically the Microcystis aeruginosa, are the dominant bloom-forming species and it is important to focus on their elimination rather than other types of phytoplankton, including dinoflagellates and diatoms. According to this knowledge, it was clear that we should mainly attract cyanobacteria of species Microcystis aeruginosa and no other species of algae to the shore. However, following our conversation with Mr. Korma, one more idea, in addition to chemotaxis, came to us regarding the potential use of modified cyanophages, viruses that infect cyanobacteria. These cyanophages would be designed to selectively infect and destroy cyanobacteria belonging to the Microcystis genus.

Meeting with Dr. Kormas

Aquatic Biologist, Professor at the University of Thessaly, Department of Biochemistry and Biotechnology.

Having two alternatives so far, we consulted an aquatic biologist for feedback on which biological solution is the most viable in freshwaters. Further, Dr Moutou, as a marine biologist, enlightened us on the eutrophication status in marine environments. While we believed that eutrophication is a major problem in marine ecosystems due to aquacultures, Dr Moutou reassured us that the majority of fish farming takes place in freshwater, since trout and salmon are the most prolific breeders in freshwater, proving that the problem is bigger in freshwaters. Regarding aquaculture, she claimed that Greece is now regarded as the leading fish farming nation in the European Union after Brexit. When we told Dr. Moutou about our idea to use cyanophages, she advised us not to deal with them because releasing them is not a safe biological solution and they are extremely difficult to work with in the lab. The optimal solution was the concept of chemotaxis, but she suggested that the chemical we will use to attract cyanobacteria to the shores must be eco-friendly and not increase eutrophication.

Professor in Environmental Microbiology and Biotechnology, Department of Biochemistry and Biotechnology, University of Thessaly

We contacted Dr. Karpouza, a specialist in environmental biotechnology, to learn more about biogenic substances and whether or not they are relevant for our project. He agreed with biogenic substances and the concept that the chemical should not affect the environment and he noted that biogenic compounds are a suitable option since they display temporal and spatial variation generating a chemical gradient in the water body. However, he highlighted that it would be nearly impossible to discover a mechanism for the modified bacteria to attach and attract cyanobacteria due to the cyanobacteria's tendency to form colonies and produce a protective sheath, which would block the procedure. Instead, he suggested that we should alter our strategy. We were recommended to focus more on the problem's fundamental cause, which is nitrates and phosphate, and to consider phytoremediation as a solution. Dr. Karpouzas had a significant impact on the project's development since he suggested we consider phytoremediation as an approach, something we had not before considered. We discovered floating wetland treatment in an article from the phytoremediation literature that he shared with us. Constructed floating wetlands (CFWs) are artificial platforms that support the growth of aquatic emergent plants. Their roots extended through the floating islands and into the water, forming thick columns with a large surface area. After reading that article about floating wetland bioremediation, we felt it would be an excellent idea to construct one carrying genetically engineered plants with enhanced nutrient uptake, in order to address the root source of the problem, the nutrient upload which leads to the extend proliferation of bloom-forming species.

Meeting with Dr. Karpouzas

Agronomist / CEO of Skoupras Agriculture Group

After rethinking our approach, the first step we made was to consult a specialist who was knowledgeable about both the problem and the plants. Mr Skoupras helped us to decide which nutrients the plant should acquire from the water body in order to combat the phenomenon via phytoremediation. First and foremost, he confirmed to us mainly that fertilizers consist of Phosphorus (P), Nitrogen (N) and Potassium (K). The fact that the nutrients present in the fertilisers may also be detected during the development of the phenomenon is evidence that the information discovered throughout our research is accurate. According to him, in freshwater lakes, phosphorus is often the primary cause of eutrophication, whereas nitrate reduction is the remedy for the groundwaters. After cross-referencing this information with the existing literature, we found that phosphorus is often the main cause of eutrophication, since 80% of eutrophication in freshwaters is restricted by phosphorus, and 10% is related to nitrogen. In an attempt to deal with the phosphorus remediation in eutrophic environments, we have concluded that it is more efficient for our plant to absorb phosphorus than nitrates.

Toxicologist, Professor at the University of Thessaly, Department of Biochemistry and Biotechnology.

Desiring to develop a more holistic approach for combating eutrophication, in addition to the excess nutrients in the water, we were also concerned about the toxins generated by the cyanobacteria of the species Microcystis in the water volume, which have been shown to have negative health consequences for humans, animal and ecosystem. Thus, we thought it was essential to get in touch with a toxicologist to learn more about these substances. Dr. Kouretas confirms that cyanobacteria species create Cyanotoxins, which are highly detrimental to aquatic organisms, wild and/or domestic animals, and people. He stated that Microcystins (MCs) as cyanobacterial toxins represent a serious threat to human and animal health and supported his argument with World Health Organization (WHO) data indicating that microcystins is an emergent public health concern. MC-LR (Microcystin-LR) is cyclic heptapeptide and is the most toxic microcystin isomer. He said that MCs are selective in the organs they damage, which is a concern for human health. According to him, the acute effects of Microcystis consumption include the destruction of liver cells, resulting in intrahepatic haemorrhage or hepatic insufficiency and persistent exposure can cause liver damage, inflammation, and, ultimately, cancer. Aware of the dangers posed by microcystins, our team was motivated to conduct extensive research to identify these toxic substances and develop a detection system for them.

This led us to wonder, how may we incorporate microcystin detection into our overall plan?

Concerning the monitoring system, only biosensors for the detection of microcystins are currently under development. Developing a biosensor by ourselves would be costly and time-consuming, so our partnership with the Manchester team helped in resolving this problem. We constructed a database of basic yet specific markers for the phenomenon, including phosphorus, nitrates and chlorophyll-α. These data assisted our team in developing a software tool that predicts microcystin concentration using artificial intelligence.
Furthermore, we imagine that our genetically modified plants could detect microcystins. It would be extremely beneficial to absorb phosphorus only in the presence of Microcystin-LR, when there is a substantial eutrophication, without interrupting the ecosystem's chemical balance.

Meeting with Dr. Kouretas

Plant Physiologist, Professor at the University of Thessaly.

While we investigated which plant would be optimal for our project, we contacted Dr. Levizou, a plant physiology expert. Based on our research of the relevant literature, we have two plant options for our design. The first was of Typha latifolia, while the second was of Phragmites australis. Therefore, we contacted Dr. Levizou to inquire whether or not the plant species we proposed are suitable for our purpose. In particular, she suggested using the Phragmites australis plants because it has appropriate features that correspond to the design we intend to execute. " Phragmites australis serves as a natural filter”, she said, as their long roots enable the absorption of the water's contaminants. In addition, she recommended P. australis owing to its extensive geographical distribution, which ranges from cold temperate zones to wetland areas in the hot and humid tropics, thus making our approach a universal platform. Microcystins, in particular, are a kind of substance that has been shown to be accumulated in our plant, as Dr. Levizou highlighted. Because of the information presented above, we decided to build our project using Phragmites australis.

Αgro-biotechnologist - Expertise: RNA-mediated gene regulation in plants.

While our first intention was to do experiments on Phragmites australis, Dr. Dalakouras informed us that, unfortunately, transformation protocols for monocot plants, such as P. australis, have not been tested enough. Therefore, we would not be able to conduct experiments on common reeds. However, he provided us with a few alternatives, including tomato plants (Solanum lycopersicum) and tobacco plants (Nicotiana benthamiana), and he suggested that we use Nicotiana benthamiana in our experiments because transformation via agroinfiltration is simple and tested before, whereas tomatoes require more equipment to be genetically modified, which would make our research more difficult. Therefore, Nicotiana benthamiana plants are chosen to be used in the Wet lab. Regarding the experiments, he said that the appropriate time for observing the leaves under a fluorescent microscope is three to four days following agroinfiltration.

Professor at the University of Thessaly, Department of Biochemistry and Biotechnology and specializes in the field of plant biotechnology.

Moving on with the development of our project's molecular mechanism and considering options for phosphorus uptake by plants via their roots, we looked into the scientific literature to investigate how plants absorb phosphorus from their roots and discovered that they use Pi transporters to do so. Dr Papadopoulou initially confirmed that it is feasible to overexpress phosphorus receptors in plant roots as well as store phosphorus in a plant's above-ground part. Our team specifically found the transporter family PHT1 (Inorganic phosphate transporter 1), a high-affinity transporter for external inorganic phosphate. Our dilemma was whether to include the Arabidopsis thaliana or the Oryza sativa PHT1 transporter in our research. She recommended that we test both the rice-derived transporter and the Arabidopsis transporter to see which one is more efficient. Thus, both transporters were included into the design of our molecular constructs. In addition, Mrs. Papadopoulou assisted us in making a decision on the repressor that should be used in our system. Repressors have been widely used in synthetic biology as they allow for a precise control of gene expression, producing a desired phenotype. After reviewing the relevant literature, we selected to incorporate two suppressors into our design: TetR and lexA. Professor Kalliope Papadopoulou advised us that the Tet Repressors are among the most common repressors that may function properly in plants. Finally, she advised the usage of the Tet toolbox, which we later use for improving our repressor with a trans regulator.

Meeting with Dr. Papadopoulou

Assistant Professor, Medical School, Department of Chemical Biology, University of Patras. - Specialty: RNA Biology

Concerning the molecular detection of microcystin in the plants, which will activate the expression of phosphate transporters, we discovered that a riboswitch may be used to bind a toxic peptide inside the root cell and change the gene expression in our system. A riboswitch is a regulatory section of a messenger RNA molecule that binds a small molecule, hence altering the synthesis of the proteins encoded by the mRNA. Aptamers are short riboswitch sequences that bind to a specific target molecule. Dr. Stathopoulos was an invaluable resource for advice throughout the project's molecular design phase; in particular, he provided giudance on the development of the riboswitch that detects microcystin-LR and regulates the repressor's expression in our system. He began by saying that designing a riboswitch from scratch is a time-consuming and independent research process. Therefore, he encouraged us to utilise riboswitches already existing in plants. Only theophylline riboswitch (TPP Riboswitch) in plants was found to be well-characterized in the literature, thus wet lab members incorporated it into the design. However, he advised us to design in silico the microcystin-targeting aptamer, which we performed. Following this information, he recommended we use the "RNAfold" algorithm to estimate the secondary structure of the new riboswitch. We found this tool to be quite helpful in our effort to develop a novel riboswitch using particular microcystin-LR aptamers.

Meeting with Dr. Stathopoulos

MSc student in Advanced Experimental and Computational Biosciences, Department of Biochemistry and Biotechnology, University of Thessaly & Member at iGEM Engineering Committee.

As a member of the iGEM 2022 Engineering committee and a member of iGEM Thessaly 2019, Nick provided us with insightful information on the modeling development of our project. After observing our initial attempt, he advised us to divide the modeling into modules, where the output of one module is the input for the next, so that the modules are connected. We followed his suggestion and created our biological model accordingly.

Environmental Engineer in the Department of Spatial Planning University of Thessaly.

Construction of CFWs may be apparently simple, but there are many parameters that affect their design and operation. Ms. Pavlineri, with her experience in CFW, warned us about some key points that should be taken into consideration in the construction of our CFW. While the plastic pipes (PVC) seemed to us like the most common and convenient material to use for the platform, during our discussion, she underlined the need to find and use renewable and more ecologically friendly materials, such as recycled plastic. The latter was the trigger for the dry lab to delve into the field of materials and discover mycelium, an emerging biomaterial that is both 100% natural and biodegradable. Moreover, she advised us of the requirement for a safety factor that is conservative owing to the increased weight that will arise from the fast growth in biomass. Specifically, she reminded us that our construction must be capable of supporting the additional weight that would result from the development of the plants over the first quarter of their lives, which will affect both its buoyancy and its stability. We have taken this recommendation into account and further developed our idea by concluding, in keeping with the circular economy vision, that the plant shoot should be harvested following bioremediation and converted into other valuable products, such as fertilizers or biofuels.

Meeting with Ms. Pavlineri

Geologist, Professor at the Aristotle University of Thessaloniki, Department of Civil Engineering.

In this stage, when we learned about a new material called mycelium, we wondered: Is it suitable for our construction? Therefore, we consulted Dr. Stefanidou, a professor with knowledge of construction materials. Dr. Stefanidou was astonished by the platform's construction material, which she claimed she had never seen before. For assistance, she recommended we give her a sample of mycelium and perform the necessary tests to measure its porosity and density in order to evaluate its suitability for the platform. Based on the measurements she conducted, we realized that we needed to pay attention to the material's permeability. As a result, we decided to cover the material with a 100% natural, waterproof protection so that our platform can tolerate being exposed to water without corroding.

Assistant Professor, School of Civil Engineering, Democritus University of Thrace - Speciality: Water resources and wastewater management

One more component we needed to add to the CFW was the phosphorus sensors we wanted to install on top of the platform to complete our monitoring system. We contacted Dr. Akratos to obtain information on the most efficient means of water monitoring. He advised that we replace the phosphorus sensors with chlorophyll-a or dissolved oxygen sensors since these are more specific indicators of eutrophication. Moreover, he brought up an important point that we had not considered. There will be a problem with buoyancy, and the platform will not be able to float and function properly if we place the sensors on it. In an effort to resolve this issue, we decided to make the monitoring system an independent component of our project. We considered creating a Remote-Controlled boat equipped with dissolved oxygen and GPS sensors to evaluate the waterbody for indications of eutrophication before implementing the CFW. This would allow us to collect precise and real-time measurements of eutrophication in sites where it is more critical to implement the CFW.

Assistant Professor, School of Naval Architecture and Marine Engineering,University of West Attica.

In an attempt to design a Remote-Controlled boat (RC boat), the dry lab department contacted Mr. Mazarakos for assistance with the preliminary model. Initially, he inspected an early design that we constructed. Following the inspection, he recommended a few tweaks in our blueprint to ensure optimal operation and safety. Notably, he suggested we lower the width of our stern (aft-most part of the boat). Dr. Mazarakos also proposed slight changes to our boat's hull after reviewing our early design. Later on, he mentioned a flow simulation software called MAXSURF. There we could assess the performance of our model at high velocity. Dr. Mazarakos also noted several factors that could setback our implementation, including the Kármán vortex street. Furthermore, he gave us valuable advice on methods we could implement to ensure our boat's material does not have high water absorbency. Lastly, he mentioned a few ways we could enhance our prototype in the future. These included generating energy through solar panels on the lid of the boat. This way, we promote sustainable and reliable energy production henceforward.

Meeting with Dr. Mazarakos

Chair of the EFSA Panel on Genetically Modified Organisms and Head of the Crop Science Department in Teagasc.

In an attempt to close the loop and get our concept to its last step, the recycling of our resources, we engaged Mr. Mullins to evaluate the feasibility of converting our genetically engineered plants into valuable products. He addressed our concerns with the recycling of the resources we use within the context of a circular economy and suggested a solution for our worries. He confirmed that the genetically modified plants can be converted into fertilizer, and although we were considering burning as a solution, he recommended composting as an alternate and more environmentally friendly biological method. According to him, composting is an absolutely safe method for the degradation of the genetically modified plants' DNA. Even if a small amount of DNA persists, it would not pose a threat since it would not affect the environment; cells and transcriptional mechanisms are required. By recycling plants that have absorbed phosphorus from freshwater and producing phosphorus-rich fertilisers, we achieve so-called zero net phosphorus, which has a positive effect on the environment, he added. Last but not least, he proposed an idea we hadn't considered before: recycling of the mycelium platform after it has been utilized to generate biomass for future energy production.

Εnvironmental Engineer, Professor at the Technical University of Crete, Department of Chemical and Environmental Engineering/ Constructed Floating Wetland Expert / European Climate Pact Ambassador / Editor-in-Chief “Circular Economy and Sustainability”

Our team decided to approach Dr Stefanakis in order to discuss with an Εnvironmental Engineer the implementation of our project in the real world. He specifically mentioned that our project is innovative for our country's status since his experience shows that CFWs are not frequently used or introduced to the public. He characterized Navanthus as a novel application of environmental engineering in Greece. Furthermore, he confirmed that the dimensioning of the construction of the wetlands were ideal, which allowed us to proceed with the study of construction’s industrial design.

Meeting with Dr. Stefanakis

Professor at the University of the Aegean, Department of Mechanical Product Design and Systems, Design and Research designer.

Considering an implementation strategy in detail, we stepped across several questions regarding industrial design and the product manufacturing process. Dr Bissas listened to our implementation plan and provided us with constructive feedback to ameliorate it. Firstly, he helped us in understanding more specialized terminology regarding the development and implementation of a product. He later highlighted the importance of having a prototype so that investors and customers can understand the design. We followed this advice by designing a small-scale platform as well as a remotely controlled boat made of the exact same materials in order for us to bring it to the competition and show them live. He noted that the product development process requires a great deal of time and labor, as well as several alternatives to challenges that may occur. In order to get our proposal closer to the actual world, he suggested that we need to consider aspects such as the cost of the materials, the suppliers and the technological infrastructure, as we documented thoroughly in the implementation page.

Meeting Dr. Bissas

Head of Institute for Biosafety in Plant Biotechnology.

Outcrossing of the GM plant is our primary concern for the future implementation of our floating platform outside of the laboratory. Dr. Wilhelm discussed with us how to incorporate plants safely in the CFW. Because Phragmites australis plants reproduce mainly via rhizomes, he said, it's vital to "cage" their roots so that they can't penetrate the water's surface. In response to this recommendation, we installed a strong, eco-friendly mesh with a filter at the CFW to enclose both the roots and the aboveground part of the plants, to prevent the spread of roots and pollen during the construction's open-release period. As extra biosafety precaution, we have considered to use a module to avoid pollination of our plant and therefore seed formation, via expression of the Barnase protein specifically in anthers. The Barnase is an RNase that, when expressed, is lethal for cells. Using this gene downstream of an anther-specific promoter, we strive to render our plant sterile. When we asked about our kill switch module, he said that Barnase expression in anthers does not have high efficiency scores, but this is not a serious concern since germination from seeds is rare in P. australis. However, we intend to incorporate the module into our project so that all biosafety concerns to be handled.

Head of Directorate, Ministry of Rural Development and Food Directorate General of Agriculture Directorate of Propagating Material of Cultivated Plant Species and Plant Genetic Resources.

When trying to solve a local and global issue such as eutrophication with a GMO solution, the legislation surrounding GMOs, and more specifically our GM Plant is vital for its future use. Mr. Kosmas was quite helpful in enlightening us on the Greek policy regarding the implementation of genetically modified plants. In particular, while many countries of the world have approved a great number of GMOs, other countries including Greece use a national ban on GMO cultivation as a measure to prevent contamination of the supply chain. Consequently, he suggested that if we aim to execute our proposal in the future, we must approach nations that permit the development of our GM Plant and comply with the international regulations regarding GMOs. After receiving this information, our team conducted research into the relevant literature and came to the conclusion that the United States, Canada, and Japan are leaders in the total number of GMO varieties approved, and eutrophication is a serious problem in these regions. Among these are the Great Lakes in Canada as well as Lake Taihu in China, both of which have high concentrations of Microcystis species during the summer months, which means that our approach might potentially be implemented in those countries in the near or far future.We also followed his advice and submitted an application for research ethics to our University Institution so that we would be legally protected during our lab experiments.

Administrator at the European Commission, Head of Unit: Sustainable Freshwater Management (ENV.C.1)

Contacting the European Union gave us important context about the regulations and legislation surrounding GMO production. Mrs. Doeser indicated that our solution comprises a GM Plant for non-food and non-feed purposes. Therefore, there is a separate legal path from GM feed crops that needs to be followed if we want to implement our approach in the EU. Referring to the specific article of the EU Environment Commission about GM Plants, we concluded that all EU countries have a strict regulation towards their implementation but the situation in each EU country may differ. It is undeniable now that we must first research the legislation of each nation before attempting to implement our idea.

Mayor of Trikala City

We had the honor of travelling to Trikala, a city located in our region, and discussed with the Mayor, Papastergiou Dimitrios, in order to discuss the phenomenon in other water bodies of the region and how the problem is being addressed by the local authorities. Specifically, he said that the river Lithaios, the river that flows through the city, shows no evidence of eutrophication and has been carefully monitored by sensors installed on the bridges erected over the river. This sensor can measure pH, turbidity, and temperature. After demonstrating our monitoring and phytoremediation system, he noted that he could see our idea being implemented as a pilot project in the city of Trikala in the future. Taking the discussion a step further, we sat together and identified on a map of the city the locations of the river's intake and outflow from the city, where monitoring can be conducted first and then CFW may be applied in case of eutrophication. The conversation with him provided us with great satisfaction since the local authorities are willing to accommodate a preliminary small-scale implementation proving the feasibility of our project.

Meeting with Mr. Papastergiou