Overview
Toxoplasma gondii is a protozoan parasite that readily infects warm blooded mammals, and is a growing contaminant as a waterborne zoonosis and increasing in prevalence in cats, with approximately 16-40% of cats contracting T. gondii within the US. Additionally, Toxoplasma gondii is currently recognized as a waterborne zoonosis, and there is prevalence in transmission to humans, sea otters, and other mammals due to contact with contaminated cat feces.
However, there is no current vaccine or drug able to be set as a definitive standard treatment for T. gondii. When an organism is immunocompromised, infection due to the parasite occurs. Symptoms often include fever, diarrhea, cough, seizures, dyspnea, and death. Furthermore, in pregnant mammals, this can cause morbidity, stillbirths, and reduced reproductive rates, and can lead to numerous health conditions or death in immunocompromised animals. In humans, complications particularly those in pregnant women include fetal death, eye and nervous system diseases, as well as numerous congenital defects.
In order to address these issues, RBHS iGEM has decided to pursue this project by utilizing bacteriophages to develop a vaccine that can be administered to cats, so that we can prevent the formation of oocysts in the definitive host and reduce the spread of Toxoplasma gondii to other species. However, to reach this point in our project, our team needed to utilize human practices. Human practices are integral in a medical and scientific community. Throughout our project, Toxoplasma Gonedii, we enquired the help of biomedical researchers, professors, and an expert veterinarian to identify a solution for our project. Concurrently, we focused on reaching out to our community to understand their needs for finding a beneficial solution to the problem caused by Toxoplasma Gondii, while also educating them about it.
Our Approach
Introduction
As Toxoplasma Gondii is a problem that impacts not just our local area but the whole world, our team decided to focus our human practices approach on creating a project that combines a community, global, and scientific perspective. Our team decided to first focus on understanding the perspective of our community regarding Toxoplasma Gondii, by identifying and evaluating the needs of our community in San Diego through surveys and presentations. At the same time, we reached out to professors, researchers, and veterinarians, to get insight about our project idea and understand our project better. To get a global perspective, we released videos, posts, and other resources that would be accessible by many people, and looked for resources that would help us understand the impact that Toxoplasma has globally.
Our team used the 5-step design process as a reference for a timeline in creating our project. This design process focuses on five steps: empathize, define, ideate, prototype, and test; however, the steps are not restrictive, and cycle back at various points. Although this process is usually used for engineering, we believe it can be applied to other scientific fields. We chose this process, because it allowed us to cycle back to previous steps to further our project.
Empathize
This was our stage of discovery, so our team focused on understanding what Toxoplasma Gondii is. We learned about the parasitic life cycle, its impact globally, current treatments being used for infection, and common hosts of the parasite. This was an important stage for us, because it helped us determine that we wanted to choose this as our project idea. We had chosen our project, Toxoplasma Gondii, because we had discovered that marine animals like sea otters were affected negatively. Through this initial stage, we also discovered other mammals who were impacted by the parasite, one of them being humans.
We came back to this stage to understand how our community would be impacted by our project. Our discussion with a veterinarian helped us understand what we should do with our project, and the importance of understanding our end-users. Our team released a survey that explained the problem, and required community insight on the knowledge they have about Toxoplasma and their evaluation of possible solutions. We utilized the information we got from them, to put in our considerations when coming up with our project's real-world application and implementation.
Define
This stage occurred concurrently with our Empathize stage. We had to decide how to pursue the problem caused by Toxoplasma Gondii. This was the point where we dived into research articles about experiments being conducted. We looked through the parasite life cycle to understand which part needed to be targeted, so that reducing the chance of transmission of the parasite would be the most impactful. We also needed to understand how we would conduct this experiment with cats in mind for receiving the treatment. Discussing with a veterinarian helped us with identifying which approaches we can take with our project, without harming cats in implementation.
Ideate
The ideate stage was a critical stage for us. It was the point where we had to decide how to approach a solution to Toxoplasma. Our team researched inhibitory compounds of T. Gondii, to see if we could find any that we could include in our experiment. We looked into other approaches, such as finding antigens, antibodies, and proteins T. Gondii that we could target for our experiment. At this stage, we needed the help of experts and researchers to understand how we were going to treat the problem. During this stage, we talked with researchers to understand which approach is the best. With their help, we were able to come up with the idea of using bacteriophages in our vaccine that would deliver an oocyst-specific antigen from a wall protein of the oocyst, to create an immunological response in the intestine of a cat.
Prototype
This stage occurred concurrently with our ideation stage. As we decided our specific approach to this problem, we needed to show proof that our concept would work. Our lab team focused on finding the right parts to have our lab work well. They were able to identify what they needed with the help of the SDSU lab students and professor that we were able to utilize the help of. Our modeling team focused on creating models to show the feasibility of our vaccine approach. With human practices, we were able to use the feedback from our survey, to understand what our future implementation should be. This feedback helped us in deciding that a vaccine system was a good approach to take with our project.
Test
The testing stage occurred after we had gotten all our lab parts to perform our lab. We were able to conduct our lab, while our modeling team focused on proving our approach was plausible. With human practices, we focused more on community interaction by using the information we learned from our survey, to bring more awareness about T. Gondii and understand what else our team should focus on to achieve our community’s needs.
Values
Scientific Feasibility
When creating a scientific project, we need to utilize the resources that we have to make our project feasible and sensible as it can possibly be. As our team approached T. Gondii, we focused on trying to educate ourselves as much as we can about Toxoplasma, so that we can make proper decisions about our project. Having an idea is possible for a project, but it has to be impactful and safe for it to be an useful idea. When approaching our idea of creating a vaccine system for reducing T. Gondii infections, we understood that there are other options as well, so we focused on establishing that it is the best way to approach this problem through various inputs. Discussions with veterinarians, professors, and biomedical researchers helped us with realizing the usefulness of creating a vaccine. Our lab team used this input to create their wet-lab approach.
We also had to understand the community’s perspective of using a vaccine. Therefore, we released our human practices survey which did ask the respondents to rate approaches to solving our problem. Creating a vaccine as a solution was highly rated, meaning that our community also believed that it was a feasible solution to the problem. Although this does mean that our project is scientifically feasible, our team also took into consideration our concerns about possible resistance of the vaccine from the parasite. However, we can not make a clear statement on vaccine resistance without having tested our vaccine at higher stages, but we can acknowledge that it is a concern.
Environmental Safety
As our team discovered T. Gondii, due to its devastating impacts on marine animals, we understood that environmental safety should be an important aspect of our project. We focused on cats, one of the most definitive hosts of Toxoplasma, as they are known to spread T. Gondii asymptomatically. As a result of run-off water mingling with cat feces that have sporulated oocysts in them, the oocysts make their way into the ocean and contaminate the water. With oocysts in the seawater, marine animals face a high risk of infection, especially sea otters. Recent findings have brought to attention sea otters dying of Toxoplasmosis. Sea otters are a keystone species in the marine ecosystem and declining sea otter populations could lead to an imbalance in the marine ecosystem, thus resulting in major environmental concerns.
Educational Importance
In a scientific field, such as synthetic biology, it is important to understand and educate ourselves about a project or topic through various resources. Our team focused on educating ourselves through various research articles and discussions with experts that brought in different perspectives regarding our project. This was impactful, as our approach to the problem caused by Toxoplasma would not have been possible without that. We understand that research is an integral part of creating a project, but it is also time-consuming, so our team has also focused on providing educational information about Toxoplasma to the scientific community through a literature review.
Throughout our project we also focused on educating people in our community. To make proper decisions about Toxoplasma, we need the perspective of the community; however, most of the general public is unaware of the parasite or the infection. Therefore, our human practices focused on several ways to provide information to our community, so that we can get their proper input on Toxoplasma. Our survey, an educational tool that we used not only for providing information to our community about Toxoplasma, but for our team to also gain input about our community’s perspective. Through proper education of ourselves and the community, our team was able to come with a project approach that respected the community’s input as well.
Toxoplasma Survey Analysis
In order to understand our community’s needs and understanding about Toxoplasma Gondii, RBHS iGEM released a survey on August 31st, 2023 and kept it open until October 2nd, 2023. We wanted to gauge our community’s initial understanding about T. Gondii and then see how they felt about our solution. Our survey was posted on Instagram, Nextdoor, Discord, and other messaging services. We hung up posters at our school and local grocery stores, so our community was able to more easily access our survey. We asked questions regarding age, gender, and education level to understand the general idea of who was answering our survey. Then, we asked three general questions regarding cats, pregnant women, and T. Gondii. After those three questions, we provided a summary of our problem, so that people are able to answer our specific project-related questions with proper knowledge. The last four questions of our survey were to understand if our community believes T. Gondii is a problem after reading and filling out earlier portions of the survey, and to acknowledge which solution they would prefer to solve this problem. We received a total of 141 responses from our survey. Our survey analysis is in the pdf below:
Timeline
iGEM Mentors
Chris Kuffner was the first person we talked to regarding our project. Chris was our team’s mentor from last year, and he helped us get started with brainstorming our project idea this year.
Takeaways: We were able to get a general sense of where to go with our project. As our team was new to wet-lab, Chris was able to advise us on which techniques are best for doing in a simple lab.
Moving Forward: Chris’ expertise helped us with identifying which lab procedures that we could do with our project. We were able to use his advice with our project with Toxoplasma later on, by utilizing the lab procedures he mentioned.
Thermo Fisher Meeting
In early June, our team was able to have a meeting with Thermo Fisher. We were invited to a Thermo in San Diego for a tour of their building and some of their labs. Our team also got a chance to present our early ideas to a panel of employees and were able to get informative feedback. Their feedback helped us decide where to go next in our season.
Takeaways: Our meeting with Thermo Fisher assisted us in deciding what our project should have, and that is feasibility and the necessity of a project.
Moving Forward: The support and feedback that the employees of Thermo Fisher were able to give on our project, were points that we were able to reference later on with our project with T. Gondii. When choosing our project idea, Toxoplasma Gondii, our discussion with Thermo Fisher helped us with analyzing it in a way to help us understand why our project was needed and to think about the end-users that would be impacted by it.
Dr. Sabina Hyland
Dr. Sabina Hyland is a veterinarian located in San Diego. Our discussion with her helped us in figuring out our end users and proposed implementation. In addition, Dr. Hyland provided us with information regarding current treatments for cats, as well as directing us in which approach to take in treating T. Gondii in cats.
Takeaways: We learned about treatments for cats with Toxoplasmosis and the general availability of knowledge on Toxoplasma. Dr. Hyland also helped us with understanding the protocol and procedures taken when a cat is infected with T. Gondii, whether it is a feral or domestic cat. She also advised us on who our proposed end users and what our implementation is used.
Moving Forward: We utilized the information we learned from Dr. Hyland, by taking into account which compounds that cats can consume, when deciding the materials for our labs. Dr. Hyland’s message on the lack of awareness of the general public about T. Gondii was a key reason for our human practices to contact our community, to understand their needs, while also bringing awareness of our problem to them.
Dr. Forest Rohwer
Dr. Rohwer is a professor at San Diego State University. We had reached out to him regarding a lab space, which he has also provided for us.
Takeaways: As an expert on viruses, he has helped our team develop the idea of using bacteriophages for our vaccine. Dr. Rohwer and his lab students have been helping us throughout our process from ideation to execution. We met with Dr. Rohwer once a week for a month, and during this time, we gave him updates on our project and he gave us insight to feasibility and modifications that we can make to our project in order to meet the guidelines.
Moving Forward: Dr. Rohwer’s insight on our project helped us with making our project meet the guidelines, with wet-lab. Our frequent meetings with Dr. Rohwer also allowed us to improve our modeling and science communication part of our project because he helped us develop the presentation we needed to make.
Nelson Vayda
Nelson is a graduate student at San Diego State University working with the Rohwer Lab.
Takeaways: He helped us greatly with our project ideation and brainstorming for our project. He assisted us by guiding our team through developing a detailed and well-thought out process for our project. He gave us insight into designing primers and making our parts because he is an expert in this field. He also gave us some guidelines into the feasibility of our project and guided us through all of our safety training and procedures.
Moving Forward: Due to Nelson’s help, we were able to have a good wet-lab. We were able to find and order the right material with his help.
Dr. Sergio Cobo
Dr. Sergio Cobo is a modeling expert from San Diego State University.
Takeaways: He helped us develop our modeling and our equations significantly. Dr. Cobo gave us more insight as to how to find some of our constants in order to better detail our model. In addition, he gave us insight into how to simplify our model and how to graph our model on MatLab more effectively in order to be able to see all of our values better.
Moving Forward: We were able to complete our model with the assistance of Dr. Cobo, and we also learned valuable modeling skills that are applicable in any field that we choose to pursue.
Discussions with iGEM teams:
Throughout our project, we communicated with other iGEM teams. As only a 2nd-year team, RBHS iGEM is fairly new. Through conversations with CCA’s presidents Weili Jin and Claire Kang, we were able to get advice in approaching our project and our human practices. Our team also met with Westview iGEM & CCA iGEM on a zoom call, where we discussed our project ideas, and gave each other feedback.
Takeaways: As CCA was also focusing on a parasite for their project, we were able to get advice from them regarding the project. They also provided us advice on how to format a human practices survey. From our team discussion, our team was able to understand the importance of making our community and the global community more aware of our project.
Moving Forward: We utilized CCA’s advice in approaching our project. As this is our first year implementing human practices into our project, CCA’s advice on the survey helped us out a lot, and we were able to reach many people in our community. We utilized the information we had gotten from the team discussion, to create global awareness about our project. We released posts, a video, and a literature review to achieve that.