Integrated Human Practice

When designing our project, we aimed to create something that could truly be appreciated in the field. We therefore explored the needs and concerns of the various stakeholders of our chosen problem in multiple ways. Each of these interactions has shaped our rather vague initial idea which was to create a smart bacteria for diagnostics.

Vidilab

Vidilab (a veterinary diagnostics company) was consulted at multiple points in our project. From the very beginning when we didn’t even have a project to the very end of our lab period in august we talked with both management (the CEO, Bitte Ljungström) and researchers at the company (Alice Anlind, Sara Hägglund Ljungström). From these discussions, we learned what the major issues of the field were and how large the need was for a new diagnostic tool for small strongyles. In the end this was the reason why we decided to take up the fight against these parasites.

Initially, our project focused mainly on small strongyles and in vitro diagnosis of the encysted larvae. From the literature we understood that small strongyles formed cysts containing hundreds of individual worms in the intestinal tract of horses. It was these cysts that we initially planned to detect and isolate from horse tissue and co-culture them with bacteria.

Vidilab was kind enough to provide access to horse tissue with encysted larvae. The cysts we received did not correspond with what we initially expected due to our previous research. Because of this, we decided to focus on detection of the worms rather than the cysts. But as we knew, free floating small strongyles do not put the horse into significant danger. Instead we decided only to focus on providing farmers with better information about their horses’ wellbeing. Due to Vidilab we also knew that the usage of anthelmintics is leading towards resistance development in the strongyle population, making them into a threat to the equine industry and horses’ physical health. By administering more accurate treatments, the resistance development rate among strongyle could possibly be decreased.

During our second visit to Vidilab they also informed us about the difficulties with detecting large strongyles since all current techniques are based on egg counting in grass samples. After some consideration we outlined a strategy to battle this problem as well. First of all we wanted to investigate the need for a better preventative tool for large strongyle infections. In order to gather the data we worked on market analysis based on a survey that we spread among horse owners in Sweden. Secondly, we wanted to try and see if a similar system as we outlined for the small strongyles could work with large strongyles present in grass, hence we made an analysis of the visibility of fluorescent proteins in grass sampling.

Interviews

Horse Owners

We used every opportunity we had at our disposal to discuss our project with real life horse owners. From these discussions we learned about the best ways of isolating samples and about the difficulty of isolating urine from a horse. We therefore moved on to investigating detection methods in feces. These interviews also first gave us the idea to start investigating on a larger scale to what extent are the horse owners aware of issues related to strongyle infections and the idea of conducting a market analysis was born.

Veterinarians

We contacted different veterinarians in the beginning of the project to see how their own experience was with strongyles. The overall view was that they did not know about horses being ill from large amount of small strongyles, but they mostly spoke of the negative effects with large strongyles. These results pushed us to investigate more on large strongyle infections.

Multinational Market Research

Before we could start with our project we first needed to look into whether there is a market for our diagnostic tool and whether we could shape the potential product to an existing niche. We were interested in the actual state of urgency of the disease along with potential concerns for the future, though as can be seen from the above mentioned communication we had been receiving some conflicting information. Some sources would state that strongyle infections are of relatively small concern whether others would see it as a large issue. To analyse this further we conducted a market analysis based on a survey that was distributed not only in Sweden but also abroad. This confirmed that the needs of the horse owners regarding large and small strongyles was consistent to Vidilabs needs, indicating that the majority of each stakeholder had the same view of the current situation. This was partly seen in the large concern regarding small strongyles, contradicting some of our skeptical sources. In addition, we used the opportunity to gather data about how a GMO based diagnostic tool will be perceived among horse owners.

To advocate our stand that the amount of treatments can be optimized, we needed to know the mean amount of treatments performed on each horse per year. We received this data as part of our market analysis. Our predictive model simulating the extension of strongyle infection dependent on the amount of anthelmintic treatments suggests that the current situation can indeed be optimized. Fewer treatments than the average today would decrease risk of developing anthelmintic resistance and still keep the horses healthy.

The market analysis gave us insight of the horse-owners perspective and the objective data that we needed to draw any conclusions. We used the data from the survey to confirm our project strategy and to build our different models on. To learn more about Market Analysis read below.

Market Analysis

What diagnostic tools are accessible today and what are their limits? Is there a market for a new diagnostic tool with our approach? Is there a need for a new diagnostic tool for large strongyles as well? How do the potential stakeholders or customer feel about using a diagnostic tool based on GMOs?

To answer our questions about small and large strongyle infections in the horse owner community we chose to conduct a survey that would query those that we intended to help with our project. Our survey was conducted to receive a greater understanding of the individual perspective. The survey was translated into 10 different languages, making it a truly international feat (see table 1 ).

Language	Amount of answers from the survey
Swedish	370
Czech	75
Latvian	38

Table 1: The table shows the translated versions of the survey with highest amount of answers.

Ethics

The purpose of our project was to combat the systematic overuse of anthelmintics [1] and thus also address the growing resistance against such compounds. Resistance may arise from multiple factors, like that of mass treatment, under-dosing and not varying the drugs used [2]. These factors may sound familiar, because this is not unlike how resistance against antibiotics arise in bacteria [3]. The resistance against anthelmintics might also be a problem for humans in the future [4] which increases motivation for a new diagnosis and/or treatment approach. This is “good” news since the organism used in our project (namely E. coli) are inherent to the microbiome of humans and horses alike. We believe that a new, specific and potentially eco-friendly method of diagnosis or treatment would have a positive outcome on society as a whole. This could lower the degree of stress and potential suffering for animals that get infected as well as for their human owners.

There are however some concerns in public about E. coli being pathogenic or a transmitter of disease. It’s often the incidence when E. coli contaminated meat for consumption gets coverage in news media. It is thus a challenge when conveying a complex topic to the public which demands some scientific knowledge, especially in biology and is something our Human Practise group has been challenged with throughout the project. The risks using E. coli and its given traits for this project are discussed further throughout the text.

GMO, Welcomed by the Public

By authoring and distributing a survey to horse owners we could see that the majority of respondents in Sweden had positive attitudes (83.2 %, 368 respondents) towards a GMO-type of diagnosis method. The survey also showed a public fear of growing anthelmintics resistance against blood worms (77.3% of 370 respondents ranked their fear as 4 or 5 were 5 being the most concerned).

Environmental Impact and Risk

The E. coli (BL-21, DH5-alpha) that was used in the lab has a biosafety level of 1, which is the lowest possible on a scale of 4 [7]. This means that E. coli is safe to work with in the school laboratories we were provided with by Uppsala University [8].

Nothing that is considered toxic or harmful was genetically introduced (UnaG, amilGFP) into our E.coli. However, for cultivation and selection purposes used in common synthetic biology practical work, antibiotic resistance was also introduced which presents a risk in the potential spread of antibiotic resistance (Chloramphenicol, Ampicillin, Tetracycline) if these plasmids were to spread into the environment. This was avoided to a large degree by following standard lab protocol with specific guidelines regarding waste disposal of GMOs.

The ability for proliferation of GMOs and organisms in general is determined by their fitness. As a rule of thumb regarding the spread of GMOs in the environment, fitness is greatly reduced for most lab strains of microorganisms because they have been modified in numerous ways to make them easy to work with. These are not adaptations that are suitable for their natural environment, giving them a negative fitness contribution [9]. Regarding the risks associated with nematodes, you can look at the separate safety form and select Uppsala from the menu.

This was the first time a team tried to see if this is an applicable solution to diagnose parasitic infections caused by nematodes in ungulates; it thus is a long way from a potentially finalized product. If a product would surface in the future, further risk assessments must be evaluated to see if an engineered product would be deemed safe to be orally ingested by a living host. Since an enteric diagnostic system based on bacteria is a novel approach there is little to be found in the literature about its applied effects. In addition to internal reviews, it is common for an external entity (like a government) to perform reviews on the product, especially if the product is classified as a GMO. This is necessary to ensure that the product is safe before letting it hit the market.

Understanding Your Limits - a Burnout Guidebook

Mental health awareness is gaining appearance in society, yet it is still a tabu in many settings. For instance, in a project that you truly care for, the risks and warning signs might go by unrecognized. It is an art to find balance at work but in our opinion it is an essential part of a success in the long run. Therefore iGEM team of Uppsala, chose to acknowledge the importance of understanding one’s limits by writing a guide-book on the topic.

The premise of the guidebook is that many iGEM teams in retrospect admit that work environment in iGEM oftentimes becomes stressful. Factors such as level of experience, responsibilities, ambitions, deadlines e.g. drive projects forward but at the same time easily lead to feelings of overwhelm and increases risk of burning out. However, even if working with iGEM underlines these factors, the topic of stress at work is recognized universally. Thereby we wanted to address this topic by creating material that anyone who seeks guidance can relate to and use. Thereby we tried to make our guidebook as universal as possible - iGEMers as well for students of any discipline and people in as well as outside of academia.

In this little book we have compiled some general background about stress, exhaustion and burnout in order to explain the issue and show that these topics deserve your attention. This is followed by suggestions of some preventive measures that we have found suitable. Some parts of the book might be a repetition of old truths but we tried to mix it up with e.g. mindfulness exercises.

In the guidebook you can also find some statistics about stress levels in iGEM specifically. These statistics are a preview from iGEM work environment report that iGEM Uppsala association is working on based on the survey described further down.

Also, check out the chapter “The importance of authenticity in Organization” from iGEM Lund guidebook . They have also been focusing on improving work environment but instead of more general depictions their guidebook focuses on hands-on-approaches for increasing efficiency through organization of work.

Our hope is that we will encourage people to not overlook the importance of their well-being and lead to healthier relation to work. As we wish to give back to the iGEM community for all the wonderful experiences we hope that future iGEM teams especially, will take part of the material in “Understanding your limits - a burnout guidebook”. iGEM is giving platform for young, brilliant people to do truly amazing things. However brilliant people don't retire after iGEM - take care of yourselves because the world needs you.