Intoduction

Our project focused on the bioremediation of wastewater which is an issue present in the industrial sector and throughout the environment. Clean freshwater is a finite resource and bioremediation of contaminated water bodies is becoming increasingly important. Emerging innovative technologies for effective and low energy treatment of wastewater exists. It was important for us to discover what impact our project could have among existing wastewater treatment technologies. We also believed it was important to obtain insight from relevant stakeholders which would help our project evolve into a valuable and useful method. In other words, we felt integrated Human Practices was paramount in the construction of our project.

Our starting point was a paper which we discovered in Nature titled: Acoustic Reporter Genes for Noninvasive Imaging of Microorganisms in Mammalian Hosts, published by the lab of Mikhail Shapiro, at Caltech. This paper used an established biological process -- the formation of simple gas-filled vesicles in known bacteria and archaea -- in an innovative way: as an imaging platform, using specific resonances to selectively collapse the vesicles and mapping their locations in mammalian deep tissue.

Through this paper, our Wet Lab team knew that viable parameters for manipulating the gas vesicle size, shape, and location did exist. These parameters exist in the form of the different genes that produce the gas vesicles; from here, we would use these parameters and combine them with the unique aspects of our project -- coupling the expression of gas vesicles with the bacteria in question binding to our molecule of interest. Now, theoretically, we could use the bacteria to separate our molecule of interest from water and then make the bacteria float using vesicles. By creating a physically separate layer where the bacteria had taken up the contaminant in question, this makes the water treatment process dramatically easier, and produces a viable bioremediation platform!

We contacted the Shapiro lab and they agreed to send us key bacterial strains needed to produce our vesicular construct. Correspondence with the Shapiro lab and previous relevant iGEM teams taught us how to optimize the parameters for gas vesicle production to meet our needs. Concurrent with this process was research and the writing of a comprehensive literature review. This research provided us with the knowledge about the genetic system responsible for gas vesicle production and that there are a few critical genes which we can be manipulated and used in the optimization of gas vesicle production, GvpA and GvpC. Conveying this knowledge to our Wet Lab Team, they were able to use these leads to optimize gas vesicle expression and create a model bioremediation system in the lab.

By developing a biomass separation platform using cells with enhanced biosorptive abilities, we can capture high-value materials or pollutants from wastewater effluent which could be a viable alternative dewatering process to complement or replace traditional centrifugation and filtration processes. Our aspiration was to develop such a platform that can be used in industry, to benefit the environment, and have real-world application. Using scientific discoveries from various articles and feedback from departmental graduate students [CLICK HERE FOR MORE DETAILS], wet lab decided to use microbial gas vesicles in the bioremediation of wastewater. Meeting with stakeholders and experts allowed us to establish our project’s applicability within the industry and the potential it had to eliminate energy inefficiencies, harsh chemical uses, expensive processes, obtainment of high-value materials and pollutant removal. Secondly, these meetings ensured our project was developed to be used responsibly. Based on the feedback received from stakeholders, public opinion, and policymakers, we had our Human Practices team focus on the applicability of our project by determining design constraints to guide our wet lab and dry lab work. We believe by focusing on addressing key issues, the rational development of a useful “GMO” technology could outweigh the often irrational fear of GMOs.

Two specific stakeholder interactions influenced our project. From this, we learned that our project had some potential to achieve remove pollutants such as microplastics and pharmaceuticals, or extracting heavy metals and other high-value materials, ultimately resulting in clean, usable water.

KINROSS - Extraction of Valuables & Removal of Pollutants

Kinross is a gold mining corporation which prides themselves on the importance of health, safety, and environment in their mining operations.

“We pride ourselves in how we in our environmental standards. We really take our approach to health, safety and the environment seriously.”
-Michal from Kinross

In mining operations, water is very important, as it is used in the grinding and leaching processes. During mining operations, Kinross implements a water strategy, which is a comprehensive program aimed at proactively addressing water issues and opportunities and aims to recycle their water at a better quality than when it was extracted.

“No waterless extraction methods available to us, all operations use water.”
-Michal from Kinross

“We actively look at reducing all sorts of contaminants in water. Our policy is to discharge water better than the condition we found the water at.”
-Michal from Kinross

Working with Kinross promotes the safe and environmentally conscious behaviour all mining industries should have. The recycling of water is financially expensive, time-consuming, and energy expensive. Our project aims to eliminate these inefficiencies and provide mining site communities and the people that live there with usable water. Thus, producing a tool that could be used for the promotion of eco-friendly industrialization.

“What you guys are aiming for is innovation, is visionary, and...Kinross can supply you with information necessary.”
-Michal from Kinross

Waste Water Treatment Plant (WWTP) - Removal of Harmful Substances from Wastewater

Ashbridges Bay and North Toronto Treatment Plants expressed their concerns for microplastics and pharmaceuticals. Safe and effective treatment of wastewater is important for sustaining the health and well-being of Toronto citizens and the environment. Current and future wastewater treatment is put under regulations imposed by the provincial and federal government to ensure the environment and public health is preserved. Our work aims to promote these standards and eliminate harms that threaten public health and the environment.

“80% of wastewater is released into the environment without adequate treatment”
-WASTEWATER: The Untapped Resource. (2017). S.l.: United Nations Educational, Scientific and Cultural Organization.

The need for a more efficient, cost-effective wastewater treatment method is essential for the environment, the global community and the progression of innovative technology in the industrial field.

Ontario Ministry of Agriculture, Food and Rural Affairs (OMAFRA)

Our interaction with the Ontario Ministry of Agriculture, Food and Rural Affairs (OMAFRA) informed us that 95% of Ontario’s food processors are linked to municipal sewage systems. This presented a direct link to our design’s applicability for Toronto’s wastewater treatment plants. Furthermore, the OMAFRA has two major issues within the food sector. The management of phosphorus due to sanitation cycles and the capture and removal of biological oxygen demand (BOD), fats, oils and greases (FOG) and suspended solids (SS). Our project would respectively focus on the removal of BOD and SS. To authenticate these applications, we met with Ping Wu, the past president at Professional Engineers Government of Ontario (PEGO). We discovered the OMAFRA had problems with BOD. As water efficiency improves in homes, due to low flush toilets, the residential BOD increases. Increased BOD results in more hydraulic capacity use in wastewater treatment plants.

In Germany and the Netherlands in the mid-90’s regulatory response to increasing residential loadings was to cut manufacturing off of sewer services. In some communities, it meant that the entire cost of wastewater treatment was then borne by residential users. In Ontario, this has happened when large food processors in small communities lose their food processors – such as the closure of Nabisco in Exeter and Libby’s in Wallaceburg.

Conclusion

We created opportunities for feedback from the public to integrate and influence the direction of our design. These included the SynBio Forum, our SynTalks podcast, and our public survey. The SynBio Forum [CLICK HERE FOR MORE DETAILS] engaged the minds of the public on the topic of synthetic biology and its environmental, legal, social and ethical implications. Audience members were asked to complete a short survey which illustrated their preconceived notions of synthetic biology and their opinion on our project. After the panel, audience members were asked to conduct a post-panel survey to measure the efficacy of the information conveyed during the panel.

Our SynTalks podcast [CLICK HERE FOR MORE DETAILS] presented the opportunity for us to gain insight from professionals from various fields associated with synthetic biology and our project. We interviewed policy experts from the Department of Environment and Climate Change Canada on the legal and ethical issues that revolve around our project and synthetic biology. Furthermore, we produced episodes which interviewed experts from the medical, agricultural, and environmental field which aimed to disclose the facts and educated opinions of synthetic biology and genetically engineered microorganisms. As a result of these interviews, we integrated the knowledge acquired from these experts and policymakers to establish how our design would be effective, safe, ethical, and beneficial.

We integrated all these aspects (interviews, meetings, outreach programs, and public engagements programs) to the design of our project. By building a relationship with Kinross Gold Corporation and the Wastewater Treatment Plants of Toronto, we were able to determine what materials or pollutants were a complication for the bioremediation of wastewater where heavy metals present in mining operations and microplastics, penicillin and other pharmaceuticals. This data was relayed to the wet and dry lab teams and was used to develop our project. Our dry lab team used this collected information to investigate the application of cellular floatation in a bioreactor-type bioremediation process. Furthermore, dry lab aimed to estimate maximum mechanical carrying capacity of an engineered biomass of a particular mass which is significant for the bioreactor model.