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In this section of our wiki, you will find precisely the role of each team member. Each member has been tenaciously working on this exciting project throughout the summer period. Moreover, we received lots of help from university staff (UNIL & EPFL) and experts in the field to develop this exciting project. Here, we show our appreciation for their contributions.

In the project description section, we have written a clear and detailed description of our project. With this outline, we expect that anyone who reads it will understand our concerns about quagga mussels as an invasive species. We also expect the reader to grasp all the issues that quagga mussels raise in Switzerland, the objectives of our project, as well as the solutions we propose to address this matter.

Within this project we pioneered the work with invasive mussels in the iGEM competition as we are the first team working with this multicellular organism. Throughout the project, we had to test if our plasmids were functional and if they would work as expected. To do so, we first needed to find the best conditions to keep the mussels alive in our laboratory, respecting ethical practices, despite the lack of details about this in literature. Secondly, we had to establish the correct way to accurately measure the effect of our engineered bacteria on the mussels' survival. We are proud to have paved the way for future iGEMers to tackle this mollusk by having developed and written several protocols from scratch about its maintenance and survival measurement techniques.

We documented the troubleshooting and optimization process in our project using the synthetic biology framework of “design, build, test, learn”. We optimized plasmids transformation into Pseudomonas protegens by electroporation and the preparation of FitD toxin samples prior to applying them to quagga mussels through several iterations of the “design, build, test, learn” framework. Using this framework, we have also succeeded in different engineering transport and catalytic plasmids for zosteric acid production. Moreover, we contributed to creating two new parts compatible with the iGEM BioBricks RFC 1000 standards to convert tyrosine into coumaric acid and for the uptake of sulfate in the cell.

We collaborated with the Chalmers-Gothenburg team to write a science communication blog: The Transcriptome. To increase the accessibility of our blog, we collaborated with several other teams to translate our articles into German, Swedish, French, Spanish, Japanese, Greek, Portuguese, and Italian. Additionally, we collaborated with other iGEM teams in Switzerland (the UZH and EPFL teams) by organizing a mini-symposium to inspire new students to join iGEM competition and to introduce Synthetic Biology to a more general audience. For this mini-symposium, we also invited PhD students (from the University of Lausanne & University of Columbia) working in synthetic biology to share their work.

To broaden our knowledge of the current methods to deal with quagga mussels, we contacted experts dealing with invasive mussels in Lake Constance, located at the Swiss borders with Germany and Austria. We met with a team of researchers consisting of P. Spaak, A. Anh-Thu Weber, L. Haltiner, and S. Rossbacher to discuss their points of view on the invasion of quagga mussels and learn their strategies to eliminate them. We also had the opportunity to meet with Nathalie Menetrey Perrottet, the Head of the Section of Water Biology at the State of Vaud in Lausanne. She shared several documents regarding the characteristics and invasiveness of quagga mussels. This information is summarized in the “Description” section.

Conducting a project in the water and sanitation sector is a very complex task, as it requires coordination of a vast array of activities, team supervision, budget management, and public communication, among other things. We designed the project, aiming to eradicate the invasive species of mussels from Swiss water services, in order to apply it to a real-life context. Therefore, we thought of a successful implementation based on consulting several experts to answer their needs and respect legislations that are in vigor.

We interviewed people affected by the mussel problem to learn what solutions they need. We talked to the co-director of La Maison de la Rivière, a foundation that protects Swiss aquatic species, and the Water Department of Lausanne. We have especially realized from their feedback that it is not only a matter of killing the mussels but also and mainly of preventing their spread. Unlike what one might think, only combining several methods of prevention, control and eradication will result in an effective change to the current situation. For this reason, after thorough research, we developed a second approach to prevent mussels attachment based on zosteric acid. Throughout these meetings, we also received multiple feedback that shaped our project, such as the proper implementation of the FitD toxin to treat clogged canalizations.

To illustrate the urgency of quagga mussels’ invasion and to demonstrate the impact of our project, we built a mathematical model based on population dynamics to estimate how long it would take for the pipes of a relevant dimension to get clogged due to the rapid proliferation of quagga mussels. Our model will be useful for water treatment authorities to choose the right pipe diameter to minimize blockage, predict the time when pipes need to be serviced and cleaned from quagga mussels' blockage, and show the effectiveness of our product (FitD toxin) in increasing the time interval before clogging occurs.

We successfully designed and implemented strains of engineered bacteria characterized by a strong efficiency in killing the mussels and identified the most potent candidate in the P. protegens strain overexpressing FitG (the activator of the FitD toxin). By calculating the survival probability of quagga mussels treated with our strains, we showed that there is a significant difference (confirmed with a Gehan-Breslow-Wilcoxon statistical test) between our FitG overexpression strain and Pseudomonas protegens wild type treatments. This is the main component of the standard treatment available on the market. A mathematical model built upon our experimental results effectively highlights the efficiency of our solution based on the FitD toxin by showing a considerable increase in the time it takes for the pipes to get clogged.

We had a productive and meaningful partnership with the University of Manitoba iGEM team. Their project focused on limiting the spread of zebra mussels (Dreissena polymorpha) in Winnipeg's waterway. Through this partnership, we shared our project ideas, which were highly relevant since zebra mussels and quagga mussels share many characteristics. Both of our teams were developing toxins to regulate mussels’ spread. We worked on the native fitD gene in P. protegens CHA0 whereas they tested two toxins: Act and a truncated version of FitD in E. coli BL21(DE3). Since we designed a reliable and working experimental set-up to test our toxin directly on the mussels, we performed this assay to test the Act and truncated FitD toxins designed by the Manitoba team. In exchange, the Manitoba team helped us quantify our native FitD toxin's protein production through our overexpression constructs.

To introduce the topic of synthetic biology to the general public, we collaborated with the Foundation of la Maison de la Rivière. We provided didactic materials about invasive species that the museum will use during its workshops. The portraits we drew up will be distributed in paper form to visitors as supporting documentation. In addition, we also wanted to create a dialogue between visitors using different media. Thus, we produced a popular science book for children aged 8 to 10, which discussed the concept of synthetic biology, environmental biology, and invasive species in the form of games, crossword puzzles, and drawings to make the learning fun, simple, and easy to understand. Through this book, we aspire to reach a younger audience that we believe is tomorrow's future. It will be distributed by the museum for an event around invasive species and used in academic classes.

Regarding the quality of our book and our involvement in educating all age levels, we believe that we have succeeded in introducing and educating the general public clearly and distinctly regarding the concept of synthetic biology, environmental biology, and invasive species. Through our efforts, we reached different age levels to raise awareness about science in general, synthetic biology, and the environmental impact of quagga mussel invasion in Swiss waters.

Our project would not have been what it is today without the invaluable help of professionals who brought their expertise and knowledge in order to conceive two distinct but complementary approaches to counter the problem we face: FitD toxin to kill the mussels and zosteric acid to prevent their attachment. Thanks to la Maison de la Rivière and the Water Department, we could discuss our product's application on a larger scale to apply it accordingly and successfully in real-world situations.
What caught our attention, in particular, was that there is no single solution but a combination of many approaches to tackle this challenge. Furthermore, one of them is to prevent mussels' attachment according to people's accurate findings on the spot. This brought us to a second approach based on zosteric acid to prevent their attachment.

We developed a novel assay to test the efficacy of FitD toxin on quagga mussels. We succeeded in developing this trial despite the lack of literature and designed it ourselves, although with the help of experts in the field. While developing our tests, we performed many experiments with very different approaches until we came up with the most relevant and effective one. As a result, we have a robust assay through which we could quantify the efficiency of our FitD toxin and the Act and truncated FitD toxin from the Manitoba team. Challenge accomplished!

Development of a model that could be used by any company and water treatment authorities working with pipes on sites with potential mussel invasion to help them choose the best diameter and service interval to avoid or slow down the damage of this phenomenon. This model was used to simulate the effect and benefits of water pipe treatment with our overexpression constructs.

We submited the following part: BBa_K4209000.