Field Background

Our team researched several project ideas and eventually settled on a biopesticide, Project Ash Guard, for the Emerald Ash Borer, due to its economic and ecological impact.

Ash trees in forests contribute to the rich ecosystem and microbial diversity⁵. These ecosystems have mutualistic and commensalistic relationships which promote biodiversity and can help mitigate global warming through carbon storing and nutrient cycling^6-8. Trees serve an important role in these ecosystems as the primary producers and creating habitats⁶. Furthermore, this loss of biodiversity from Ash trees can result in invasive plant species taking over forests and other habitats where Ash trees would be if not for the infestation⁹.

The Emerald Ash Borer (EAB), also known asAgrilus planipennis (A. planipennis), is a highly invasive, non-native species, first documented in Canada in 2002^10,11. The adult EAB (order Coleoptera) feed on Ash tree leaves and then their larvae feed on the phloem and other tissue under the Ash tree bark¹¹. Due to their inward feeding, it is hard to detect EAB infestation in the early stages¹¹. This leads to tree mortality and eventually reduction of ash tree populations¹². As a result, EAB infestation is both destructive and economically significant¹³. It has been estimated that there has been over billions of dollars in economic loss from the EAB in Ontario¹⁴. In Canada, it has been estimated that the cost of replacing both urban and rural Ash trees would be $1.422 billion¹⁵. This ash loss is estimated to persist for decades¹⁶. Furthermore, Canadian ash is part of the Canadian forestry industry, which contributes roughly $20.9 billion to the Gross Domestic Product (GDP), and thus ash damage from the EAB has a negative impact on the quality of life of Canadians¹⁷.

There is further economic loss through trade restrictions to prevent the spread of the EAB and monitoring of these “EAB free” zones¹⁵. The Canadian Food Inspection Agency (CFIA) oversees the regulated areas¹⁸. As of 2019, EAB infested areas include Winnipeg (Manitoba), Ontario (Sault Ste. Marie, the Algoma District and Thunder Bay), Quebec City (Quebec), Halifax (Nova Scotia), and New Brunswick (Oromocto)⁹.

Figure 1. Emerald Ash Borer infested areas in Canada. Arrows highlight regions in Canada that the Canadian Food Inspection Agency has reported to be infested with the Emerald Ash Borer.

This loss of ash tree population can also cause health effects^19-21. There is elevated risk of diseases caused by air pollutants as a result of infested trees not filtering the air²¹. Ash trees can also prevent heat-related deaths¹⁹.

Despite the gravity of the Emerald Ash Borer, there are few treatment methods. There only exists 1 pesticide approved for usage in Canada against the EAB which may only be used on healthy trees^9,22. The chemical methods of prevention however these do not decimate EAB populations during an infestation so the infested trees must be cut (removed) and burned which will contribute to greenhouse gasses and global warming²². Several areas including the City of Guelph have removed Ash trees to prevent widespread infestation²³. If there are no Ash trees, then an infestation cannot occur, however this harms the ecosystem^5-8. Scientists have explored the option of biological controls including parasitic wasps and natural pathogenic fungi, however these methods are still in development^9,24

What can be done to treat EAB infestations without destroying Canadian biodiversity?

Initial Project Design

The initial project design was sculpted through an extensive literature review. The initial project idea was presented to our Primary Investigators (PIs), Dr. Stephen Seah and Dr. Rebecca Shapiro. Out of the ideas presented, Dr. Seah thought that our Emerald Ash Borer (EAB) biopesticide (now called Ash Guard) was the most feasible option due to available research and our previous experience with a biopesticide project. In 2022, we worked on Project Ceres, a biopesticide for tomato greenhouse growers²⁵. We could use the research and past insights from our 2022 project to optimize this year’s project.

Our main focus for our initial project design was to have a functional concept and then we could modify it to better solve the EAB problem and minimize the downsides of existing treatments^9,22. After literature review, the wet lab team found that the toxin Cry8Da impacts the Japanese Beetle and some products that stated that the toxin impacted the EAB^22,26. There was minimal research proving the claims of toxicity against the EAB, so the team decided that it would be a good project topic.

Would it be possible to improve existing methods of treatment for the EAB? Could consumers have more options? Would it be possible to stop an active infestation?

Figure 2. Novel idea for a bioinsecticide for the Emerald Ash Borer. The Cry8Da protein from Bacillus thuringiensis can be inserted into a plasmid which can then be inserted into E. coli for biomanufacturing of the protein or Saccharomyces cerevisiae can have the Cry8Da plasmid inserted into it so that the yeast strain will produce the protein.

Project Refinement

Throughout our project, biweekly (as in every other week) PI meetings have been integral to refining our approach. During these meetings, our executive team presented project updates and questions/concerns to both of our PIs for feedback and guidance. One significant suggestion was the use of Saccharomyces cerevisiae (S. cerevisiae) as a vector by Dr. Emma Allen-Vercoe, a researcher at the university who provided guidance for our 2023 project. For further insight Dr. George van der Merwe, a Professor and researcher on campus that uses S. cerevisiae was reached out to for his expertise. Dr. Van der Merwe has helped the wet lab team by providing insight on yeast plasmids, providing yeast plasmids (which were transformed into E. coli) and the S. cerevisiae strain.

Our team has weekly wet lab meetings with our Project Leads (PLs) for research updates and weekly executive meetings to ensure continuous progress and collaboration. Exec would also meet on a weekly basis to discuss project updates and concerns which led to the improvement of our project.

More details about Project Refinement are on the Engineering Success page.

Introduction to Stakeholder Work

Our Human Practices work consisted of 2 parts: project related outreach and the 3D printing project outreach.

The business/human practices team was introduced to a brief overview and project framework which would serve as useful tools and a foundation for the team’s human practices and stakeholder work. This presentation can be accessed through the link below:

Project Framework Presentation (PDF)

In the first official human practices meeting, we introduced our business team to stakeholders and stakeholder communication in a presentation presented by our Human Practices Lead, Kamal. This presentation went over how to determine stakeholders and how to communicate with them, which is through catering your message to each stakeholder.

We briefly did an initial identification of stakeholders through a Value Proposition Canvas and Business Model. The Value Proposition Canvas and Business Model can be seen below:

The Value Proposition highlights the benefits (“value”) that our project would deliver to our stakeholders and potential users. This value is a novel bioinsecticide that would hopefully combat the disadvantages of existing solutions (expensively, timely) while effectively targeting the invasive Emerald Ash Borer turn protecting/preserving the Ash tree population without resorting to widespread Ash removal and thus protects animal habitats and ecosystems.

The Business Model represents 9 categories necessary for a business’s survival and includes many topics that are important for introducing stakeholders to our project. It was created using the presentation below:

Business Model Canvas (PDF)

Image from the University of Guelph

We received feedback and advice from Erin Young, Business Incubator Services Manager, on how to conduct stakeholder communication and possible directions/resources for our stakeholder and field research.

Some key points were:

Image from the University of Guelph

Another expert at the University of Guelph we reached out to was David Hobson, Manager at the Research Innovation Office. In his role as Manager of Technology Transfer & Entrepreneurship, David Hobson helps researchers with protecting their Intellectual Property (IP) and commercializing their work²⁷.

Some key points were:

Image from the University of Guelph

Erin Young had connected us with Nakita Byrne-Mamahit from Project Soy Plus. In Project Soy, Nakita is the Coordinator from the Research Innovation Office. Project Soy Plus is a student competition that focuses on sustainability and projects that relate to the Sustainable Development Goals (SDGs)²⁸. It is a great opportunity for student groups such as ourselves to explore the implications of our project and further develop it, slowly moving towards potentially commercializing our project.

Some key points were:

Through these initial stakeholder meetings, we’ve discovered a direction for our human practices work which is further explored through our Stakeholder identification map.

In our project we believe the following Sustainable Development Goals (SDGs) are emphasized:

Target 15.1: “By 2020, ensure the conservation, restoration and sustainable use of terrestrial and inland freshwater ecosystems and their services, in particular forests, wetlands, mountains and drylands, in line with obligations under international agreements”²⁹.

By protecting Ash trees, our bioinsecticide aims to help conserve and restore terrestrial ecosystems. Ash trees are an important part of biodiversity as they provide a vital habitat for wildlife and help with important ecosystem roles such as carbon sequestration and aiding in microbial biodiversity.

Target 15.5: “Take urgent and significant action to reduce the degradation of natural habitats, halt the loss of biodiversity and, by 2020, protect and prevent the extinction of threatened species”²⁹.

By halting the destruction of the Emerald Ash Borer, its destruction of Ash trees and the habitats Ash trees are a part of, our bioinsecticide supports the preservation of biodiversity by safeguarding the Ash trees within Canada.

Target 15.8

Our bioinsecticide aims to reduce the impact of the invasive, non-native Emerald Ash borer and mitigate the damage of the beetle, which threatens forests. Our team’s actions helps reduce the impact of the Emerald Ash borer and helps support the eradication of the beetle from infested areas.

As we look at implementing these targets, it is essential to communicate and work together with stakeholders to find a balance between their views and values, and our own - conservation, minimal harm, sustainability, resource management and more. Also, it is essential to answer questions about which regulatory body will oversee our project, ensuring accountability and adherence to federal standards and legislation.

Read more about our human practices below.

Project Feedback & Help

Throughout our project, we sought input from key stakeholders, including Entomologists, Government officials and agencies, pesticide users and field experts. Their feedback provided valuable insights and have raised important questions which have helped refine our approach and ensure that the project aligns with both our and their values. This collaborative feedback is crucial for the success of our project.

Entomologists

Dr. Steven (Steve) Paiero

We had reached out to Dr. Stephen Marshall (Professor Emeritus) in the School of Environmental Sciences, who had previously worked with the EAB for information about the EAB and if he knew where we could source EABs from. Since he is retired, he referred us to Dr. Paiero the Curator of the Insect Collection at Guelph.

Dr. Paiero provided us with advice for how to find EABs in Guelph, as we are legally allowed to collect beetles in our area, and went on an excursion on campus to look for EAB infested trees with our Director of Research and some of our wet lab project leads. While this outing was not successful for finding EABs, we found some trees with signs of EAB infestation and he provided us with a list of other possible areas on campus for us to look at. He also referred us to 2 government scientists who have worked with EABs in the past, however unfortunately they were not currently working with EABs.

Jake (Jacob) St. Amour

A member of the wet lab found a Government lab (Great Lakes Forest Research Centre) that sold Emerald Ash Borer beetles to labs looking to do research with them. We connected with Jake from the lab who shared their lab’s care guide and more details about the beetle, afterwards our team ordered some beetles (30 of which 15 were female and 15 were male) from the NRC Canada laboratory and they were so kind to ship some adult beetles as we had not realized that typically they ship out ash bolts which would take almost a month before they are out of diapause and ready to test on.

A human practices member, Sarah, had followed up with Jake to see if there was anyone he knew doing research related to pesticides or biopesticides that he could connect our team with.

Pesticide Users

Alison Morrison - University of Guelph Arboretum

Additionally, we reached out to the University of Guelph Arboretum Manager Alison Morrison who is a certified Arborist and has sufficient experience in caring for and treating Ash trees affected by EAB infestations. As a consumer of current commercially available treatments for Ash trees she was able to provide us with insight into the feelings of consumers and the necessity for a product such as Ash Guard.

Currently Alison and her team use a product called TreeAzin which is an insecticide however, they are backing off application rates. Reducing the use of current treatment options is related to the rate and procedures for applications causing too much damage to the trees prior to clearing the EAB infection. Consumers are seeking cheaper options that cause less wounding to trees and have the ability to be used long term. Additionally, current treatment options for Ash trees were designed primarily to slow infections and create enough time for cities to effectively remove trees and not to clear the infestation and prioritize tree longevity

With this knowledge the value of a product such as Ash Guard becomes evident. Alison suggested that catering our product to be cost effective and have an application method which does not wound the trees would be most effective.

Protein Biochemistry Expert

Dr. John Dawson

We had also reached out to Dr. John Dawson who is a professor and faculty director at the University of Guelph who has extensive experience with protein biochemistry. He recommended that we look into other biological vectors that are more specific to the host. The Cry8Da protein has the potential to harm native beetles in Ontario that are beneficial to the ecosystem such as the bronze rich borer beetle (Agrilus anxius) and the two-lined chestnut borer beetle (Agrilus bilineatus). He recommended that we use computer software such as AlphaFold to study the binding of the Cry8Da protein to the midgut lining of the EAB. With a deeper understanding of the chemical reaction, it may be possible to enhance the Cry8Da host specificity to the gut lining of the EAB through genetic modification, decreasing its effectiveness on other native species of beetles.

In terms of turning the pesticide into a spray, Dr. Dawson recommended that we research the effectiveness of polyethylene glycol in preventing the degradation of the spray and maintaining its liquid form. Once we are comfortable with the spray design, he recommended multiple product tests including testing the half life of the spray, testing different Cry8Da concentrations on the emerald ash beetles, and doing multiple environmental exposure tests to see if the spray can withstand UV radiation, heat, cold, and Canadian weather conditions

He encouraged us to keep an open mind about our design and look into other ingestion methods other than a spray, and for ways for the pesticide to spread amongst the population after it is initially introduced. Such as an Emerald Ash beetle releasing viral cells after death for other EAB to consume.

Government Officials

Timea Filer

We reached out to Timea Filer, an Urban Forestry Field Technician from the City of Guelph natural resources sector to better understand governmental responsibilities in the utilization of insecticides. The use and legalization of insecticides in Canada is regulated by Health Canada’s Pest Management Regulatory Agency (PMRA). The primary product currently approved for use in Canada is TreeAzin, however it is applied to the trees by injection and causes significant wounding to the plants. This product was primarily produced and approved to maintain tree health for a period of time until municipalities had sufficient time to remove all infected trees. TreeAzin, unlike Ash Guard, is not intended to cure EAB infested trees but to treat symptoms.

Despite the overarching regulations by PMRA, the responsibility of insecticide use is placed on municipalities and individual property owners. The government branch responsible for specifically the regulation of the EAB infestations and communication with municipalities on treatment of infected trees falls to the Federal Government Canadian Food Inspection Agency (CFIA).

Regulations and Implementation

First explored in our meeting with David Hobson, regulations pertaining to synthetic biology which falls under biotechnology and genetic engineering in Canadian legislation, is a critical step in bridging the gap between our research project to its real world application. Health Canada typically oversees the regulation of biotechnology, however other agencies (when applicable) have the joint responsibility for overseeing the delivery of the biotech products³⁰>.

As David mentioned, our project may fall under the jurisdiction of several government agencies, so we reached out to Health’s Canada Pesticide Management Regulatory Agency and asked if our project based on our proposed implementation would fall under their agency’s governance or a separate agency. As of September 20th, we are awaiting a response.

While waiting for a response from the Pesticide Management Regulatory Agency, we decided to look into governing legislation through a google search.

Pesticide Management Regulatory Agency/Health Canada

The Pesticide Management Regulatory Agency (PMRA) is a group under Health Canada which oversees the approval of pesticides in Canada under the Pest Control Products Act³¹. Their authority also falls under the Canadian Environmental Protection Act, 1999 which is jointly upheld by Environment Canada³⁰.

The PMRA offers a free pre-submission consultation to applicants for them to gain guidance prior to submission. This consultation is required for Microbial applications, which our project would most likely fall under, and Joint review requests³². This guidance covers study protocols and data so that applicants can submit a high quality application^32,33.

Figure 1. Pesticide Management Regulatory Agency (PMRA) pre-submission form.

The main criteria that must be met for pesticides in Canada are³⁴:

1. Human health risk assessment: The Canadian population’s health is considered, however there is a special emphasis on the health of fetuses, infants and children who are going through developmental changes that may be more susceptible to chemical hazards from pesticides³⁵>.
2. Environmental assessment: All pesticides must not pose environmental concerns and if it poses unacceptables risks to environmental health it will not be registered for use in Canada³⁵.
3. Value assessment: This assessment determines the pesticide’s contribution to pest management including efficacy, impact on host organism, and health/environmental benefits³⁴.

The procedure for pesticide approval is a lengthy process to ensure that pesticides are safe for the environment and human health, which is backed by the PMRA group re-evaluating pesticides every 15 years to ensure that there are no adverse effects and the approved usage is still appropriate³¹.

While the federal government oversees approval of pesticides, the municipalities oversee usage and they have bylaws for the usage of pesticides, which is formed with insights from the Ontario Pesticide Act and federal legislation^35-37. The provincial government may restrict or prohibit pesticides in their jurisdiction and the by-laws from municipalities can further restrict pesticide usage³⁵.

Environmental Protection Agency

There is a similar governmental body to the PMRA for the United States (US) called the Environmental Protection Agency (EPA) which regulates pesticides and ensures their safety³⁸.

The EPA registration process the following is required³⁹:

Figure 2. Environmental Protection Agency (EPA) pesticide submission form.

This process is rooted in 2 major American Laws: the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and the Federal Food, Drug, and Cosmetic Act (FFDCA). The FIFRA requires pesticides in the US to be registered under the EPA and the FFDCA requires a set tolerance level for pesticide residues used in or on feed³⁹.

Following the Canadian pesticide regulations listed above are crucial for the successful implementation of our product. Understanding and being aware of these regulations is the first step in ensuring our compliance. Our team will prioritize these regulations as we continue to research and improve the insecticide, ensuring that it meets all legal and safety standards.

There are several possibilities of implementation methods of the insecticide onto ash trees. One promising option is developing a spray using the Cry8Da protein, with E. coli used to mass-produce the protein. This would allow for efficient, large-scale production of the active protein ingredient. However, transforming the product into a spray raises important considerations, including shelf stability, storage temperature, and the duration of its effectiveness.

Another approach is utilizing a yeast strain with potential insect-repelling properties which could be applied onto the ash trees in the form of a spray or a topical agent in order to protect them from the EAB. Considerations with this method include ensuring the yeast’s target specification, and determining the frequency of application needed in order to protect the ash trees.

There are similar products to our insecticide currently on the market, such as the Vestaron peptide-based insecticide made by Spear⁴⁰. This product targets common crop pests including aphids, spiders, mites, thrips, and whiteflies⁴⁰. The company uses large biodegradable peptide molecules to target pests effectively with minimal harm to mammals, birds, fish, honeybees and other organisms that benefit their environment.

In terms of implementation, Vestaron’s journey started with researching and isolating spider venom peptides. After their research was found significant, they incorporated the company and seeked regulatory approval from American regulatory agencies such as the EPA for their SPEAR®-T and SPEAR®-Lep insecticides to be used comercially. After approval, a headquarters office was opened in North Carolina, and the product was launched in the US. After a market was established in America, the company then expanded to international markets such as the European Union and Mexico⁴⁰. Researching this company has provided valuable insights into the implementation plan of our eco friendly insecticide.

Hardware Design

The Engineering team created a 3D printing database for researchers to modify lab equipment to be more accessibility friendly with designs tested in our iGEM lab, which can be viewed in detail on the Hardware page. This page will describe in length the Human Practices of the project, and ensuring input from students and researchers this project would most help.

This project came to fruition as students with disabilities are underrepresented in STEM due to inaccessible lab equipment and the team wanted to overcome this accessibility barrier^41,42. There are many unknown barriers and known barriers that students with accessibilities face in the lab such as inadequate accommodations, difficulty using equipment due to physical conditions (task specific barriers), and inaccessible laboratories due to architectural design^41-44. With the help of the team’s free 3D database, researchers around the world can download the 3D printing design files so that they can modify their lab equipment to meet their accessibility needs.

To better address the accessibility concerns that researchers/students in labs with disabilities face, the Human Practices team created a survey (“Accessibility in the Laboratory”) to gather feedback from the labs on campus to help us identify issues and develop solutions that would make lab work more accessible for everyone. This feedback would then be implemented by the engineering team to design 3D printing designs which would be printed using plastic from recycled polypropylene pipette tips and then tested in our iGEM laboratory, which would help determine the functionality and if the designs needed to be further edited.

The questions included covered background of the individual, general and specific accessibility questions and voluntary contact information for additional feedback. See the full survey questions below:

iGEM Guelph - Accessibility in the Lab Survey (PDF)

This survey was shared over our Instagram and our university’s sub-reddit. Our Instagram post reached 2,104 accounts (as of August 15th) with 83.2% being non-followers and on Reddit we received 3.6k views of our post.

Some concerns within our survey included:

Our team discussed some other ideas between exec and leads with lab experience. Some of our ideas included:

Through the Engineering team’s 3D printing project, sustainability as envisioned by the Sustainable Development Goals (SDGs) is promoted⁴⁵. The following SDGs are used within the Hardware project:

SDG #9 - Industry, Innovation & Infrastructure

Target 9.4: “By 2030, upgrade infrastructure and retrofit industries to make them sustainable, with increased resource-use efficiency and greater adoption of clean and environmentally sound technologies and industrial processes, with all countries taking action in accordance with their respective capabilities”⁴⁶.

The hardware project exemplifies sustainable innovation by integrating recycled material into the 3D printing process. The plastic waste produced in labs yearly is estimated to be 12 billion pounds⁴⁷. Through transforming used pipette tips (a common lab waste usually made from polypropylene) into functional research tools, this initiative reduces lab waste and sustainable practices within the lab^47-49.

SDG #10 - Reduced Inequalities

Target 10.2: “By 2030, empower and promote the social, economic and political inclusion of all, irrespective of age, sex, disability, race, ethnicity, origin, religion or economic or other status”⁵².

The hardware project promotes social inclusion of individuals with (hidden or visible) disabilities within the scientific community. By providing accessible tools designed for researchers with disabilities, the project breaks down physical barriers to participation and systemic inequalities that limit the involvement of students and researchers with disabilities^41-44. Furthermore, these designs will help prevent researchers from developing musculoskeletal disorders, which 60% of researchers develop due to ergonomic strain from laboratory activities⁵³. With the designs in this database, all individuals regardless of their abilities can contribute to science and participate in a diverse and equitable research environment.

SDG #12 - Responsible consumption and production

Target 12.5: “By 2030, substantially reduce waste generation through prevention, reduction, recycling and reuse”⁵⁴.

Responsible consumption and production is a key facet of the project through transforming a common laboratory waste product, used polypropylene pipette tips, into research tools^47-49. This project reduces the environmental impact associated with lab waste disposal and sets a standard for sustainable development through prioritizing environmental stewardship⁵⁵.

SDG #13 - Climate action

Target 13.3: “Improve education, awareness-raising and human and institutional capacity on climate change mitigation, adaptation, impact reduction and early warning”⁵⁶.

Likewise, this project reduces the carbon footprint associated with producing new materials and lab equipment through creating them with recycled plastic filament made within our own lab^47-49. It raises awareness about the role of innovation in mitigating climate change⁵⁵.

By transforming plastic waste into a valuable resource (recycled filament), our project reduces environmental waste and pollution while promoting a circular economy. This project paves the way for more eco-friendly manufacturing solutions for common lab wastes.

Education & Outreach

Throughout our project we aimed to raise awareness about synthetic biology and our project. This fulfills Sustainable Development Goal (SDG) #4

SDG #4 - Quality Education

Target 4.4: “By 2030, substantially increase the number of youth and adults who have relevant skills, including technical and vocational skills, for employment, decent jobs and entrepreneurship”⁵⁷.

Through hands-on experience and training, iGEM Guelph equips its members with skills in synthetic biology, biotechnology, scientific research, the application of business to synthetic biology, engineering experience, scientific outreach and making STEM digestible for the average person, and web design and coding skills. The team equips members with employable skills and skills in emerging industries. We also foster teamwork, innovation, and critical thinking, which are crucial skills for employment.

Target 4.7: “By 2030, ensure that all learners acquire the knowledge and skills needed to promote sustainable development, including, among others, through education for sustainable development and sustainable lifestyles, human rights, gender equality, promotion of a culture of peace and non-violence, global citizenship and appreciation of cultural diversity and of culture’s contribution to sustainable development”⁵⁷.

Through our 2024 projects, we have emphasized the Sustainable Development Goals. Team members have gained insight into sustainability through our engineering team’s 3D printing project. Through our education projects, we have emphasized global citizenship and quality education to help increase scientific literacy and comprehension. Furthermore, in partnership between our education and wet lab work we have conducted outreach pertaining to climate action and the importance of conservation, and how our bioinsecticide helps with conserving Ash trees and biodiversity.

A detailed description of our education work can be found on the Education page.

Final Reflection: Challenges & Future Directions

When reflecting on Ash Guard and our 3D printing project, several challenges and potential future directions exist:

Biopesticide project

1. Challenge: Regulatory Approval - Obtaining regulatory approval for biopesticides may be a lengthy process as they must meet stringent safety, efficacy, and environmental standards. Navigating this complex landscape is essential to bringing the biopesticide Ash Guard to market.
2. Challenge: Project Implementation - The successful implementation of Ash Guard will require coordination and cooperation with various stakeholders, including governmental agencies and conservation organizations. Also, educating the public about proper distribution and usage of the product is imperative.  
3. Future direction: Increasing specificity - A possible future direction would be enhancing the specificity of the biopesticide to minimize non-target species. This precision would safeguard ecosystem health by enabling native species to thrive. For more insights, read the Future Directions page.
4. Future direction: Ecosystem interaction - As our target implementation of the bioinsecticide would either be a protein based spray using E. coli to mass produce the protein or a yeast that inhabits the Ash trees to prevent infestation by the Emerald Ash Borer (EAB). For safe implementation, we will need to do further studies (in lab) that investigate the environmental conditions needed for our protein to achieve optimal toxicity and its interactions with a mimicked environment to ensure that its use remains beneficial to overall ecosystem health without unintended consequences.
5. Future direction: Scalability of the biomanufacturing of the Protein - Adding the Cry8Da protein to E. coli enables fast biomanufacturing of the protein, and to improve this biomanufacturing the team could investigate optimizing the production of the protein using industrial fermentation and other biotechnological techniques.  

3D printing project

1. Challenge: Material consistency - One of the challenges in developing 3D printing filament is ensuring material consistency. Pipette tips can vary in plastic composition and as such we have focused on using one type of plastic (polypropylene) but a future direction for the 3D printing project could be expanding the type of plastic used to maximize the impact that recycling common lab waste has.
2. Future directions: Partnerships and Expanded Applications - Future plans could explore partnering with local labs and recycling companies to expand the 3D printing project and further reduce lab waste. These partnerships would be vital to grow the project’s impact and ensure sustainable labware.

Conclusion

In conclusion, our human practices work was rooted in addressing real-world challenges through thoughtful engagement. From navigating regulatory approval to tackling implementation of our biopesticide (Ash Guard), we remained committed to creating a sustainable solution to the highly invasive and destructive Emerald Ash Borer (EAB), which prioritized Ash tree and ecosystem health. Our 3D printing project further highlighted the importance of recycling lab waste and circular economies. Our education projects promoted quality education and scientific literacy. Moving forwards, we hope to continue projects that prioritize the Sustainable Development Goals (SDGs) and focus on community and ecosystem health and wellbeing.

Documents

See below for a copy of email drafts that our Human Practices team used to reach out to our various stakeholders.

Human Practices Emails (PDF)