Since the beginning of our journey with iGEM, our project has come a long way with many adaptations, improvements and implementations. Building a project from scratch has been challenging for all of us and required constant evaluation, reflection and patience in order to serve all of our values that we want to address with our project. During our time in iGEM from the beginning to the end, we prioritized gaining input

Let’s go through our Human Practices Agenda!

Our Human and Practices Agenda

Figure 1: Our Human and Practices Agenda

To us, iGEM has been an enormous learning experience where we learned from each other to present a multifaceted skill set that we all have in our mind.

Serving our values!

First and foremost our project deals with an endeavour that has been known ever since the discovery of antibiotics. Alexander Fleming said: ”The time may come when penicillin can be bought by anyone in the shops. Then there is the danger that the ignorant man may easily underdose himself and by exposing his microbes to non-lethal quantities of the drug make them resistant.” [1] The world has come to a point where we develop new and change known antibiotics to conquer microbial infection just to face resistance against said antibiotic. There is a group [2] that has observed microbial resistance in a matter of only 11 days. A common threat is an infection by methicillin-resistant Staphylococcus aureus (MRSA). It was first reported in 1960 and can be treated with common antibiotics. Yet, the first known resistance was discovered one year after MRSA was reported. [3, 4] The following generations are facing a new danger, the generations before us never pictured. Infectious diseases that were once easy to treat are now life-threatening.

Why did we develop B.L.I.S.S? When we began our journey, carving out the paper strip as our flag ship, our main goal was to protect human health. However throughout our iGEM journey and many outreaches, we realized that B.L.I.S.S. is much more than that. The benefits range from health, environmental, hygiene to business and ethical aspects. This has led to the emergence of B.L.I.S.S. as an environmental, ethical and health-friendly model that can be perfectly implemented in biotech companies, highlighting the potential of our project. We are sure that with our commitment, we have laid the foundation for the reliable, inexpensive and accurate detection of antibiotics in our wastewater.

HOW CAN B.L.I.S.S HAVE A GOOD IMPACT TO THE WORLD?
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Environment

The “clean” water released from the wastewater treatment plant, will eventually end up in the river, where animals and plants reside. Such antibiotic resistant bacteria that could interfere with the ecology of the river, can therefore pose a serious threat. Apart from that the flowing water distributes the resistant bacteria along a long route contaminating soil [5]. Here antimicrobials can have an endotoxic effect on the soil reducing microbial diversity which can hinder the protection against antibiotic resistance. [6]

How do we counteract that?

According to Singer et al 2016 [7], minimizing the dissemination of antibiotics prior to their discharge into the natural environment is a major step towards protecting our environment. By directly contributing to clean water that should be free of antibiotics, we also protect the environment at the same time. By directly targeting the wastewater treatment plants, the point where the removal of antibiotics and antibiotic resistant bacteria is crucial, we facilitate the detection of antibiotics to move the spotlight to the root of antimicrobial resistance. Such clean water can then improve environmental health by preserving microbial diversity in soil while also maintaining balance in the aquatic environment.

Human Health

According to the World Health Organization, antibiotic resistance represent the biggest global threat endangering human health and wellbeing [8]. The overuse of antibiotics blur their effectiveness contributing to a growing number of infections that are hard to cure and directly increase mortality. In fact Jim O’Neill stated that by 2050, 10 million will be attributed to AMR [9]

How do we counteract that?

By protecting our environment we simultaneously protect human health which presents our project as a multifunctional and multitasking project with many benefits affecting each other. Water is the most prevalent and important fluid, humans, animals consume and is an important factor for photosynthesis [10]. In other words, every living being dependent on the availability of clean water. To prevent the presence of antibiotics and resistant bacteria before the consummation, a reliable detection of antibiotics is crucial as means of preventing the onset of deadly infections.

Water Hygiene

Water hygiene is of everyone’s interests. In order to make sure that we do not not take up antibiotics and resistant bacteria that contaminated our drinking water it is of uttermost importance that we have techniques enabling the detection of contaminations in water. Low income and middle income countries suffer the most from the burden of antibiotic resistant infections [11]. Inadequate water sanitation can greatly exaberate this increasing the death rate even further. This aspect can be directly linked to human health and environment which sets our project into a complex network.

How do we counteract that?

With our paper strips consumers can make sure that their water is not contaminated. Not only would this make drinking water safe but also without any concerns and fear. With bioinformatical modification of the binding pocket within the receptor of our two component system and docking experiments, we make sure that besides β-lactam antibiotics, other types of antibiotics can be easily detected. Since clean water is also directly associated with food supply including agriculture, clean water is essential for workers and customers in market and for reliable food processing. [7]

Accessibility

Our product should be accessible to everyone. It is a privilege to be a scientist, having access to a laboratory and (funnily) be able to use all the equipment. As a scientist, we can collect water probes from wastewater, the environment and even my home and test them with elaborate and time consuming protocols. The pandemic has shown how easy it can be to perform scientific data acquisition with a strip of paper. This was one of our inspirations. What if everyone in a hospital can test the water for antibiotic pollution? What if you could do the same at home?
The work is not done by knowing if there are traces of antibiotics in the water. We wanted to develop a monitoring system for the places where it is needed the most: at wastewater treatment plants, the meat industry and hospitals.

Safety

The European Union is strictly against introducing genetically modified organisms into the environment [12]. We had to find a safe and functional solution and were surprised when we found it in bubble tea. The bubbles in bubble tea are made of alginate. It is a hydrogel that is also used to deliver therapeutic drugs in mice. Another question we posed: Can it entrap bacteria? And yes, it can. We can encapsulate our genetically modified bacteria in alginate and read out the detection via the fluorescence emitted by a protein, that is only present when the antibiotic of choice is also present (thanks to our bright minds who have designed the plasmid encoding a system that leads to the transcription of GFP). The application of the paper strip does not require the bacteria to touch the water source. A few drops of the water probe are enough for the color change.

Ethical Considerations

Antibiotic resistance is a concerning topic for ethical analysis as the consequences for human health and well being are severe but the harms of infections are often inequally distributed and highly depend on human actions such as discarding antibiotics.

How do we counteract this?

The overuse of antibiotics has ethical dimensions including food and environmental aspects that need to be addressed. There is often misuse and overuse of antibiotics by humans which increased morbidity and mortality. Our model requires no specific equipment, the paper-strips are similar to a COVID-19 test or a pregnancy test where you also see a color change. In this way, no one is being neglected and our tool available for everyone. We want to encourage people to consider how they discard antibiotics. We try this by updating our audience weekly on our Instagram account with random facts about antibiotics.

Reaching out to a variety of Stakeholders

We quickly noticed that Human Practices will be the core of our entire project thereby introducing the Human Practices cycle. It first greeted us during brainstorming. How will our idea impact health and the world? Will it really benefit consumers? Who should we talk to?

Amid the implementation of our ideas into the wet lab and dry lab we considered feedback from experts helping us to ensure reliable and promising results. Moreover stakeholders from many different kind of fields gave us feedback thereby helping us to further improve our project considering how our model can be expanded beyond water hygiene.

In our meetings we have met interesting people who haven given us valuable insight on the current situation of the issue. We did not know what it was like to be part of a wastewater treatment plant or how severe the overconsumption of antibiotics really is. It is worse than we expected.

Overview of the diverse categories of stakeholders that we reached out to

Figure 3: Overview of the diverse categories of stakeholders that we reached out to

Outreaches

Let Us Guide You through Our Journey.

WHO Flag

According to the WHO, antibiotic resistance is increasing in a concerning rapid manner and is considered to be the leading global threat affecting human health, food safety and environment in the 21st century [8] . As a result our ability to treat bacterial infectious diseases might become ineffective since antibiotics will lack effectiveness.

  • more than 670.000 antibiotic resistant bacterial infections in Europe [13]
  • 33.000 people dying in the European Union from resistant infections [13]
the global burden of antibiotic resistance is comparable to that of influenza, HIV/AIDS and tuberculosis combined. Robust investments and interventions to address this threat are urgent and would have a beneficial impact on the health of the whole world population [8]

Jim O’Neil

Since their discovery in the 1940s, antibiotics have revolutionized medicine as people were finally able to recover from fatal diseases such as tuberculosis, syphilis and much more [14]. However due to the overuse and misuse of antibiotics, they have become victims of their own success! Alexander Fleming, the discoverer of antibiotics warned the population during his Nobel prize speech in 1945 emphasizing the risk of resistance:

“It is not difficult to make microbes resistant to penicillin the laboratory by exposing them to concentrations not sufficient to kill them and the same thing as occasionally happened in the body”[15]

Nowadays the misuse of antibiotics have decreased the effectiveness of antibiotics. In his review on antimicrobial resistance, Jim O’Neill states a very concerning statistical analysis on antibiotic resistance:

"By 2050, antibiotic resistance could take up to 10 million lives each year."[9]

These numbers were without exception concerning to us but also motivated us to find a sustainable yet simple solution as soon as possible!

Throughout our project, we were able to meet different stakeholders with various professional backgrounds. In sum, all these meetings led to the establishment of a diagram summarizing all the concerns that arise from the accumulation of antibiotics in our wastewater [1].

Overview of all the concerns and issues that arise from the exploitation of antibiotics, its outcome and other factors that contribute to this global threat

Figure 6: Overview of all the concerns and issues that arise from the exploitation of antibiotics, its outcome and other factors that contribute to this global threat

Together with the stakeholders, we come to one conclusion: The Presence of antibiotics in wastewater is directly affecting the environment and human health and a robust way to prevent this issue is needed until it is too late!

One of the main questions that we would ask our stakeholders would be of course: “Do you think our model could be beneficial in detecting antibiotics?” “Do you think we could have an impactful contribution in stopping the progression of antibiotic resistant pathogens?”

Many of our stakeholders gave some valuable opinions but also suggestions on how B.L.I.S.S. could be a part of protecting humanity from antibiotic resistant pathogens.

Dr. Minh Thu Nguyen stated that it is important to target the root of the problem: the accumulation of antibiotics in wastewater! This is in line with our propose, as we are convinced that the major cause for the emergence of multiresistant pathogen is the presence of a pool of antibiotics in the first hand!

“It will have to be converted in the next 10 years” - Mrs. Neblung

Of course we sought out wastewater treatment plants and asked for their opinion as we aim to incorporate our alginate capsules into the system of wastewater treatment plants to enable reliable detection of antibiotics under constant flow. Therefore we talked to various professionals from this field to get more insights into how water is cleaned and how our paperstrips and alginate capsules could contribute to water hygiene.

Mr. Mathieu from the wastewater treatment plant in Heusenstamm was delighted when we introduced our project as it is an easy and cheap way to detect antibiotics. Not only do our paperstrips and alginate capsule help to contribute to clean water but it also saves a lot of space which was one of the issues why no technologies for detecting antibiotics could be incorporated into the treatment plants.

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WWTP

One major advantage of our paperstrips is that one can test on-site and does not have to send it to laboratory. Therefore our paperstrips are an easy way to detect antibiotics in wastewater without the need of any expensive lab equipment.

“Your paperstrips are easy to use and seem extremely affordable.” - Mrs. Neblung

Our paperstrips and alginate capsules could pave the way to protect the environment as we would prevent the spread of resistant pathogens within the aquatic fauna which was also suggested by Ghazi Akbar! All the positive feedback aside, we also received a number of suggestion that could level up our product. It gave us the impression of real interest from everyone we have talked to. For example, Dr. Minh Thu Nguyen reminded us that it is important to be able to detect certain concentrations of antibiotics. On top of that our paperstrips should be as sensitive as possible as water could contain only traces of antibiotics. Conny Neblung and Tobias Mathieu from the wastewater treatment plant were also interested in the lower detection limit of the paper strips. Dr. Anastasia Lev stated that, our paperstrip are a versatile way of detecting antibiotics! It can be adapted to several types of antibiotics which we have confirmed through our docking experiments. Moreover our paperstrips could be applied to other sources where antibiotics might accumulate such as dairy products. Just like Anastasia Lev, Mr. Mathieu suggested to add more parameters to our paperstrips to save even more space and time and to test different concentrations of antibiotics within our wastewater. Conny Neblung (WWTP Nordhausen) as well as Tobias Mathieu (WWTP Heusenstamm) have reminded us that the paper strips have to be accepted as THE method for the detection of antibiotics in wastewater. So far there are kits for the quantification of chemical oxygen demand which are commonly used in wastewater treatment plants. Mr. Mathieu had another striking idea to simplify testing even more: Have different biosensor on one paper strip. We really liked both inputs.

During our talks with many stakeholders we wanted to know whether our paperstrips are in a way accessible to everyone. It is our aim, that anyone including those with a low-income but still are interested in testing their own water, and those with little scientific background can operate our paperstrips.

There is no need for any laboratory or expensive equipment to detect antibiotics in wastewater, This can not only safe time and space but it can be used by anyone” - Ghazu Akbar

Mrs. Neblung also confirmed the simplicity and accessibility of our approach as our paperstrips do not rely on any complex laboratory equipment. Our paperstrips operate in a familiar way likewise to a COVID-19 test or a pregnancy test. Therefore it won’t be difficult to develop a product that works in the same way but for the detection of antibiotics. It is important that especially people from low-income countries have access to the paperstrips as they are above all mostly threatened by multiresistant infections. Therefore our paperstrips, that can be handled by anyone could be a large step in providing people in needs the ability to protect themselves from such diseases. In our interview with Ghazi Akbar we came to the conclusion that our paperstrips might be an easy way to provide people the ability to test their water for antibiotics, since it works in a familiar way to the COVID-19 test. Dr Anastasia Lev also pointed out that we should include a liquid like hydroxyperoxide that can inactivate the immobilized bacteria on the paperstrip once we discard them. This would further increase the safety of our paperstrips in a way that anyone can use and operate them without any concerns. Apart from that, Dr. Anastasia Lev stated that our paperstrips are extremely versatile and that we could take advantage from it by expanding its capability by detecting antibiotics in dairy products.

“Prevention is the best form of public health!” - Samuel Ujewe
“Humans, especially in low income countries are linked to the contaminated aquatic environment through their drinking water.” - Ghazi Akbar

Throughout our interviews with Ghazi Akbar and Itir Gydirici we were reminded by the fact that low income countries are even more seriously affected by the progression of multiresistant pathogens. This is due to the people getting their source of nourishment from the river for example during fishing and people bathing and washing themselves within the rivers while being exposed to multiresistant pathogens. As a consequence, people in low income countries will be exposed to a higher risk of death which is aggravated by the lack of accessibility when it comes to individual and unaffordable treatment options in these countries.
Regarding that Dr. Samuel Ujewe underlined that our project is good in two ethical dimensions:

  • We are giving access to people to test their water so that they can receive clean water. At the same time, no one needs to worry about expenses as we aim to make the paperstrips inexpensive and efficient
  • While one multiresistant infection is arising in one part of the world, the people living in that corner will have a way of keeping these infection within this circle of the world. This is possible since our team could provide them tools to already fight such multiresistant pathogens in that area before it is spreading to other parts of the world. Having this in mind, this would help all the people in the rest of world as they would never have to worry about suffering from multiresistant infections.

One of the main problem is the misuse of antibiotics that leads to the progression of multiresistant pathogens. In order to motivate the population to prevent the progression of antibiotic resistant pathogens it is important to raise awareness. As Dr Gordon and Mrs Neblung already stated, it is important to decisively address the government as they have an important influence on the establishment of new regulations when it comes to antibiotics. One major step to reach that is to raise awareness about antibiotic resistance and reaching all the people who contribute to the accumulation of antibiotics in our wastewater. This can be achieved through promoting education on antibiotic resistance.

Closing the loop!

B.L.I.S.S Is ...

SAFE

Our Alginate capsules are a safe way of detecting antibiotics in our wastewater. This is due to alginate being biodegradable and biocomatibel without producing any toxic side effcts. Mooreover no toxic solutions are needed to test our water with the paperstrips so that people can screen their water for antibiotics without any concerns!

A CONTRIBUTION TO RESEARCH

Our tools to detect antibiotics has been praised by many of our stakeholders. The next step would be to find a way to filter out multiresistant pathogens by directly targeting them. This is the start of a journey to stop the progression of antibiotic resistance! Besides the multiresistant pathogens it is also important to find ways of filtering out antibiotics within our wastewater: After detection comes depletion of antibiotics! This again shows that our project is the foundation for many more research projects!

ACCESSIBLE AND EASY TO OPERATE

Given the issue with lack of space and time in wastewater treatment plants that is hindering us to detect antibiotics inside the wastewater, our paperstrips and alginate capsules could be a great compromise. Not only is there no need for any laboratory device, but it is also extremly easy to operate by anyone as our paperstrips work in a famillar way as the COVID-19 Test or the Pregnancy test.

ADAPTABLE

This especially applies to our paperstrips. Not only could they be able to detect multiple concentrations of antibiotics in an extreme versatile way but their potential could be expanded by enabling the detection of antibiotics in dairy products and meat. This shows that we built the foundation to a product that can be further expanded and improved to help protecting the environment.

GOOD FROM A BIOETHICAL POINT OF VIEW

Through its simplicity and accessibility B.L.I.S.S. is giving each individual even those from low-income countries to protect their health. However it is important to directly confront everyone who is misusing antibiotics. They are the source of the accumulation of antibiotics in our wastewater. If we sensitize them and the government we could make a huge step forward

PROTECTING THE ENVIRONMENT

Thanks to Professor Kempf and Ghazi Akbar we learned how our environment is affected by multiresistant pathogens. By directly reducing the presence of multiresistant pathogens in our wastewater, we are also protecting the aquatic ecologic system. As the aquatic system is directly connected to all the other ecologic system B.L.I.S.S. can directly contribute to protecting the entire environment.