(Integrated) Human Practices

In working through our designs, our team has prioritised the feedback and suggestions offered to us in order to uphold the safety, efficacy, and security of the project. We developed Bo-Find to help communities environmentally and economically, as the cattle industry is a major part of life in our area. Our hope is that Bo-Find can aid those in the cattle industry by offering simple and accessible testing for Bovine Respiratory Diseases (BRDs). We acknowledge that there are potential risks to improper use or consequences gone unconsidered, however. That is why, as a team, we have done all we can to reduce and eliminate any possible negative impacts certain parts of our project might create. To do this, we have engaged with several stakeholders and experts regarding the cattle industry, cattle research, and previous solutions to the growing issues of BRDs and subsequent ARGs (Antibiotic Resistance Genes) amplified by the mass treatment of cattle herds. We have also consulted our local agricultural community, and sought the advice of multiple individuals and organisations, through the Tech Futures Challenge, BioTreks, and more.

During the first few weeks of project development, the team was split into two groups so we could divide our focus between our two project ideas, L.evothyroxine and Bo-Find. Our project idea, L.evothyroxine, was based on developing a new form of drug administration for hypothyroidism. Our second project idea was Bo-Find, a detection kit for antibiotic resistance genes using loop-mediated isothermal amplification (LAMP).

To help us kickstart the initial development process, we participated in the MindFuel Tech Futures Challenge (TFC). This event is geared toward High School and University students developing projects using synthetic biology that challenge issues in the local community. Through TFC, we were also able to access resources and a mentor (Ethan Agena) who could help us along our project development. At this point in the TFC cycle, we made video pitches for both of our projects. Our hope was that we could gather enough feedback to choose which project was better suited as a focus moving forward.

During the project pitch phase, our team was awarded Best Pitch - High School for L.evothyroxine. Some of the judges’ feedback on L.evothyroxine was largely positive, though it included comments on whether our solution would be cost effective and whether this project would be feasible with our time and understanding. We also learnt that, in dealing with hypothyroidism, we must be careful, as overdosing could lead to the extreme opposite of the current issue: hyperthyroidism. This led to further ideation about possibly developing a food-activated killswitch in order to keep the thyroxine levels ideal.

The feedback on Bo-Find was equally positive, and we received suggestions on how to ensure the sustainability of our device, and regarding the next steps of development. One judge posed questions on the actions that would be necessary to actually use the device, such as how the DNA sample would be extracted from the cattle as well as how common false positives are with the use of LAMP technology. This, later on, led us to consult Dr. Tim McAllister, (as seen below) to learn more.

With feedback collected for both projects, we decided that we would choose a direction for our whole team to follow. After a team vote, we initially decided to pursue L.evothyroxine. At the time, with the research that had been done, it seemed to be the best and most interesting choice of project. However, after diving further into the subject, we discovered that overcoming the obstacle of thyroxine’s lack of solubility would prove to be very difficult, especially within the iGEM time frame. It was at this point that our team came together to decide on whether we wanted to modify the L.evothyroxine project or pivot to Bo-Find.

In the initial steps of Bo-Find, we spoke with Dr. Kim Stanford, a leading cattle researcher at the University of Lethbridge. She applauded our desire to design a new detection platform for the cattle industry, providing suggestions on which avenues of detection have been attempted previously and how our team could go about our project in the iGEM time frame. One point she made concerned the specificity of our detection kit. To assure the effectiveness of Bo-Find, Dr. Stanford suggested we focus on the most important pathogens associated with bovine respiratory diseases. This would allow our detection kit to deliver the greatest impact for producers. Thanks to her extensive knowledge of cattle research, our team advanced our initial conception and narrowed down our focus to BRD detection.

We also discussed our initially proposed LAMP (Loop-mediated isothermal amplification)-based detection kit with Dr. Tim McAllister, a cattle researcher from Agriculture and AgriFood Canada. Knowing the benefits of other detection methods, Dr. Mcallister suggested looking into what has been done with BRDs previously. This led our team to pivot to using a different isothermal method called RPA (recombinase polymerase amplification). RPA is more suited for our purposes, being faster and more simple to use for the general population. So, our team decided to employ RPA going forward, thanks to Dr. Tim McAllister’s professional insight into the project.

We reached out to several local livestock vets and connected with Dr Steve Hendrick from Coaldale Cattle Health. We asked him about how cattle producers normally monitor their livestock. He told us that a "cattle health attendant or pen checker" walks through the pens to identify any sick cattle. Sick cows are removed from their pen and taken to their treatment hospital or an isolated pen. Animals suspected of having BRD will have their temperature taken and recorded along with their body weight. These parameters are entered into animal health programs to track these diseases. The data is analysed for any trends that may indicate an outbreak may be developing. Often 10 - 12 samples is sufficient to give an idea of the organisms involved in an outbreak. Usually some bacterial culture, serology, or PCR in some outbreaks is used to get a better handle on the organism(s) involved. Some of these might be isolated from swabs or lung tissue samples collected from mortalities.

Dr Hendricks emphasized that the biggest fear for producers might be finding a pathogen of human health concern (zoonosis) that is reportable to the government (such as Salmonella, etc). However, most producers would welcome this type of rapid testing. The biggest issue has been speed or throughput of these assays and making them simple enough to use at a vet clinic.

"We would all like to have chuteside diagnostic tests that could tell us which animals have a pure viral infection as we would be less likely to treat the animal with antibiotics. Having a better idea of the organisms at play would help us understand these outbreaks better and come up with new or better vaccines." - Dr Steve Hendricks

We visited Amino Labs, a company of experts in accessible genetic engineering education, and they gave us advice about making a prototype of our device. This included suggestions on what functions to prioritise and ideas on how to get a working prototype faster. Their advice was invaluable, and made our team aware of multiple aspects of development that we had not considered. Furthermore, we were advised by the team at Amino Labs about designing certain aspects of our device, such as using a heating element block that had adjustable temperature settings, what battery pack to use (or indeed, if we even want to use a battery pack), a possibility of implementing technologies from PCR machines, among other things. The team also discussed with us how to create a more sustainable/reusable device, and we also covered the possibilities of making the device colour-blind friendly, or using an app to notify the user of the results of the test.

Following the Amino Labs visit, we learned about the many stages of the development of such a device, and we had determined a few important guiding factors that would help us design our prototype: we needed a small, reliable, and portable device ; this device needed to be reusable; this device needed to be cheap; it should be simple to build and use; we would use a PTC heater and digital temperature controller that would heat the device to 37 and 100 degrees celsius; and that the PTC heater would be required to heat an aluminium holder block.

The Amino Labs visit also helped us begin to prioritise what aspects of the device we wanted to focus on: we realised that though we had very high ambitions about what this device would be able to do, they were finer details, and we needed to focus on getting a genuinely working prototype first. We also decided to prioritise making the device as inexpensive as possible, easily useable by anyone, and making it reusable (meaning it had to withstand the large temperature changes repetitively), and making it sustainable by building the finished product out of as many re-used or green products as possible, such as the possibility of using eco-friendly and heat resistant plastics to build the shell of our device.

After incorporating the feedback from our TFC mentor, cattle and technology experts, we built a minimal viable prototype for the TFC prototype challenge. More details on this process can be found on our hardware page. We traveled to Calgary, Alberta to the prototype showcase to discuss our project and won “Most Iterative Award”.

Along with the award, we also received advice from the judge’s reports that we endeavoured to incorporate into our project. This included a warning to be careful of the heating unit with our electronics and the materials used for the kit (see modelling). One judge also wondered about the interest level of farmers for our device and who would be our target audience. Given that BRDs present a large economic issue in the cattle industry, and our device is meant to be used to diagnose and allow for immediate action to prevent their spread, as well as the information we have gathered through reaching out and visiting with involved local organisations, our team has assured that Bo-Find will benefit and therefore interest our target audience, those in the cattle industry as well as veterinary practitioners. Overall, our experience with the MindFuel Tech Futures Challenge and all the feedback we received through them was very helpful in advancing our project.

Our team also wrote a paper through BioTreks, an international synthetic biology journal, authored and reviewed by high school students. Through BioTreks, our project was judged by multiple professionals, who then gave us their insight through comments and questions. One key takeaway included being cautioned to be aware of possible DNA contaminations, which we took into consideration during the testing of our project. Another piece of information we received was that it's possible that our reaction efficiency is affected by potential reduced assay sensitivity. The heat treatment technique is useful, but we learned through this feedback that it may relate to reduced DNA collection leading to less accurate results. As well as this specific feedback, we were given information on proper testing methods, accurate testing instructions, and sustainability, all of which we were able to address in further iterations. Additionally, they raised points about the heating mechanism and energy sourcing for the device, mainly concerning our battery-based approach. Because of this, we modified our design to allow it to function in the most effective way possible. These were the main insights we were given, but overall our project was well received. Because of our BioTreks participation, we were able to further our project and consider alternative methods.

Our team went to visit the Lethbridge Research Centre at their open house. Dr. Tim McAllister was our guide on a bus tour during which we observed crops, research buildings, and cattle. Afterward, we went into the barn. There, we saw the controlled environments for the animals, different measuring methods for greenhouse gases, and the centre’s current research on BRDs. We also learned about how the scientists battle against methane and gas emissions in the atmosphere. One presentation explored different types of media for BRD pathogen identification, and how others have fought against BRDs and ARGs (antibiotic resistance genes). We found this very helpful, as it directly relates to our project, and went forward in implementing this information after the visit. We also met Cheyenne Conrad, whose documented research directly helped with our BioTreks paper and project design. Overall, the visit was a great learning opportunity, as we left more informed about the environmental research going on locally.

In the initial stages of developing Bo-Find, our team began with the goal of creating a hand-held, rapid detection kit for ARGs. This then pivoted to BRDs, due to the diseases’ local economic impact, including their effect on ARGs’ bioaccumulation because of mass antibiotic treatment. We aimed to prioritise efficacy, simplicity, and efficiency, doing what we could with the limitations that arise from the device’s size. This, of course, was a lengthy process, including consulting multiple professionals and organisations, as seen above.

Due to the nature of our project, and how the device is meant to be used by individuals, there is always the possibility that it could be misused. To prevent the accidental misuse of our detection kit, our team has focused quite a bit on ensuring the design is simple and understandable, since incorrect use of the device could lead to incorrect results. Along with the actual model of the detection kit, our team also made sure to add a control tube along with the four others used for proper identification. This will allow the user to check that the device is working as it should.

Bo-Find, as a whole, is intended to aid the agricultural, and more specifically cattle industry/research communities. As such, we have consulted with multiple individuals and organisations in order to further inform ourselves on the subjects. As we acknowledge the presence of current testing methods for BRDs, our team expects that our project will not replace these other detection methods. Because our project aims to be simple and portable, it is not going to be as specific and thorough as the tests performed in the labs. As such, we expect these other methods will still be used for more accurate diagnoses, while Bo-Find will allow for prompt action in the field.