Team:Duesseldorf/Human Practices

Human Practices

As biology has a safe place in our hearts, our mission is to spread awareness about nature and to strive for a sustainable environment for all living organisms that we share the world with. This is much more than research and lab work - biology means future! Since the very beginning of the project, the team was interested and decided to work on phosphate recovery from wastewater. Phosphate is an essential element for plants and thus for the production of fertilizers. As it’s a limited resource, ending up in groundwater, rivers and lakes, causing for example algae blooms and eutrophication, we aimed to close the phosphate cycle. If we could solve this, the use of phosphate would be way more sustainable and we won't ever run out of phosphate and save our planet.

A survey for farmers

Since the goal of our project is a sustainable phosphate additive for fertilizer, it is only natural we would like to know the opinion of people working in agriculture, who are most likely to use our product. Our team members made the effort to contact acquaintances running a tree nursery or local farmers to conduct a survey.
In the following we list the questions we asked and the summarized replies:

  1. Would you use a fertilizer that contains a genetically modified moss? Please justify your answer.
    Here the responses varied. The tree nursery would not mind using such fertilizer as long as there were no consequences due to the modification. They even mentioned they occasionally use moss in their compost, which they use as fertilizer. The farmers on the other hand were much concerned about genetically modified elements coming in contact with their crops and possibly with their livestock, as that violates regulations.
  2. What are your precautions to pretend eutrophication?
    Both sides unanimously said they do not fertilize more than necessary and pointed out the regulations regarding this question. As growing crops is their livelihood they care about the environment and its sustainability the most.
  3. What criteria are crucial for you, when you buy fertilizer?
    This question was also answered unanimously. First and foremost is the ratio of price to performance. The performance is of course measured by the ratio and amount of nutrients, without any artificial additives. To compete with mass production in foreign countries, a local, sustainable fertilizer should come close to the same ratio. If that was the case, many would prefer the local option due to environmental reasons.
  4. Have you ever encountered a phosphate deficiency in your fields?
    The tree nursery had this issue a few times in the past and it was solved by changing the soil or targeted fertilization. The farmers on the other hand never had that problem. Some actively avoid any deficiencies by fertilizing based on nutrient samples they take on their fields.
  5. Do you change fertilizers from time to time and if so, why?
    Everyone who took the survey mentions they keep a close eye on the market and try out alternative fertilizers from time to time but usually they stick to the ones they are using.
    Some farmers even admitted they are very conservative in that regard and would need a really valid reason to switch products.
  6. Feedback
    Overall our project peaked interest as it was praised as an “easy” and interesting solution to eutrophication. The farmers appreciated our engagement for environmental sustainability and they hope our idea sparks more public awareness for the topic.

Wastewater facilities

Looking for some expertise on how to apply our product in wastewater treatment we turned towards various regional facilities. Unfortunately many facilities were not responding to our repeated emails. Only Duesseldorf’s wastewater facilities were responsive and answered a few sample questions via mail.

We learned the facility predominantly uses the chemical precipitation for phosphate using iron or aluminum salts. In one facility they additionally use phosphate elimination to increase the precipitation. Generally they easily surpass the minimal requirements.
Describing our project, without lab data to back it up, and thoughts on application proved difficult via email and due to Covid-19 a visit could not be arranged.

In case we choose to carry on our project in 2021, further cooperation with Duesseldorf’s wastewater facilities will prove to be indispensable in optimizing our product.

Social Media

Our team was most active on Instagram this year. We came up with our GMOnday, where we post pictures of our mascot Physco, who shares interesting facts about GMOs with the public. Our goal here was foremost to change perspective of how GMOs are perceived by the public.
Additionally we shared similar posts on special occasions. For example on world oceans day we raised awareness for ocean pollution and what each individual can do against it.

Fig. 2: Our mascot Physco sharing facts about GMOs on Instagram
Fig. 1: Our Instagram story for the oceans day on the 8. of June, 2020

Integrated Human Practices

Getting started - a meeting with our professors

In the beginning of the year, when we were still researching to finalize our project, we organized a meeting, where we intended to present our ideas to professors and doctors affiliated with iGEM. They were Prof. Dr. Axmann, Prof. Dr. Weber, Prof. Dr. Pauly, Prof. Dr. Schmitt, Prof. Dr. Zurbriggen and Dr. Dumpitak. We hoped their experience from years of research would help us expand our view on the project.

At the time our team was set on working on wastewater remediation using cyanobacteria or microalgae and our targets were heavy ions, pesticides and phosphates. As expected there was a ton of feedback that came our way, even some things we had not considered before. Unanimously was the notion our project was way too broad and not distinct enough. For one, the lab work and research for the treatment of different kinds of pollution in wastewater was way too labor intensive for the timeframe iGEM provides. Secondly our project had nothing to sell it on. We lacked anything innovative to compete with. We decided to shift our focus on phosphate only, as it is partially responsible for eutrophication and as such is a pressing topic in our home region, Nordrhein-Westfalen, with its high population density and strong agriculture. From there we tackled what we could do with phosphate that is innovative and useful to water remediation. One of the first ideas was a type of phosphate sensor in a chassis of our choice but we wanted to recycle the phosphate. So we put the sensor idea by side for a potential side-project, in case things go really well in the lab and we want to extend our project. (Without knowing we would not have access to the lab this year).

At this point Prof. Dr. Weber suggested being more open-minded with our chassis and proposed moss over microalgae or cyanobacteria. Its advantages included being an uncommon chassis, compatibility with bioreactors and being easy to harvest. It also provided the possibility of basic research in the go-to moss chassis Physcomitrella patens in terms of cloning, transient expression and phosphate metabolism. The idea was very well-received and intriguing to everyone. After a little extra input we assembled the final project proposal of polyphosphate accumulating moss - Mossphate. With that we had something we were confident in to be an eyecatcher and our overall project was finally visualized.

Feedback from farmers

The feedback that we got regarding our fertilizer described in our Human Practices page caused us to think more about the economics of our project, as well as the feasibility of a GMO-based fertilizer. While we feel that a GMO approach is a safe and effective strategy for increasing the phosphate accumulation in P. patens, current legislature is not favourable and farmers generally prefer a safe route. To circumvent this problem while still creating a strain with high phosphate accumulation, classical breeding and mutagenesis techniques can be used to great effect, creating a non-GMO, high phosphate strain. This is not our preferred method, as mutagenesis can have unforeseen effects on the plant. For the european market and other areas with such restrictions, this is likely still the better solution, even if overall we believe it is a less effective solution than targeted engineering of P. patens.

Above all, the farmers valued effectiveness, so creating a cost effective product is our priority. This is reflected in the design of our bioreactor, which is used to optimize growth times and make the drying process as easy as possible, making Mossphate an economically viable product. Read more about it here.

Juelich - Expertise in bioreactors

The more we visualised our project, the more we realised we needed a system to apply our moss efficiently. If a moss was to be used for water remediation, it would have to to be in conjunction with some sort of bioreactor. However as moss is not a common chassis to begin with and no team member had experience with bioreactors, we had no idea where to start. Luckily our university is in close relationship with research centre Juelich and they regularly support Duesseldorf’s iGEM teams. Juelich’s department for alternative biomass uses a broad variety of different photobioreactors and therefore they were the ideal contact for our problem.

The head of the department, Dr. Holger Klose, was our first contact. He provided us with the first outside perspective of our project and pointed out crucial criteria for a reactor utilizing moss. With him we discussed different types of reactors and their up and downsides. Additionally he asked fundamental questions to our project such as why we use moss over cyanobacteria and what precautions we plan on taking in regard to GMO laws. Dr. Klose’s feedback helped us to focus the application of our project on the agriculture sector. He also supported us with further contacts within and outside of Juelich. One of those was a startup company named “Phytolinc”, who specialize in culturing microalgae on custom membrane-photobioreactors. We contacted Dennis Prausse, one of the company's founders, and had an extensive phone call about their systems and how it could be applied to our moss. Although we did not end up using their system, the exchange of opinions helped us understand the necessary properties our envisioned bioreactor needed.

The other crucial contact shared by Dr. Klose was Ladislav Nedbal, who works at Juelich in the plant science department and specializes in bioreactors. At this point in time we had thought more thoroughly about what type of reactor we were planning to use and wanted to present the ideas to Mr. Nedbal. Instead Mr. Nedbal presented us a system we had previously not considered at all - the turf scrubber. It is not commonly used but the system seemed ideal to build as a prototype, while also fulfilling many favoured attributes we envisioned. Following the interview we put all our effort into designing and building a functional turf scrubber prototype.

Fig. 3: Skype call with Dr. Holger Klose

Acquiring our moss - Frommer and Bezanilla lab

We were lucky enough that the Frommer Lab at our university was working with Physcomitrella patens and we just had to message them and ask for it. Manuel Miras from the Frommer Lab was happy to provide us with a sample of the strain, so we were able to start propagation ourselves. He also provided us with the contact for the Bezanilla Lab where he got the plasmids he used for his experiments. After contacting them as well they permitted us to use those plasmids. However in the end we decided to work with the plasmid pShdpy provided by our advisor Anna Behle instead.