Our team took the safety implications of our project seriously, and thus considered various factors to ensure the safest conditions for our project. During the course of the season, our team utilized Escherichia coli strains BL21 (DE3) and DH5a. These strains are both non-pathogenic strains that are classified as BioSafety Level 1 (BSL-1) (REFERENCE). Additionally, our team also used two fungal species, Sclerotinia sclerotiorum ,Pestalotiopsis microspora . These are both non-pathogenic to humans and also classified as BSL-1 (REFERENCE). Though the fungal species are spore-forming, we completed the additional safety Check-In form as required by iGEM. Our lab is also a certified BSL-2 lab, allowing us the required certification to work with the strains of E. coli and the two fungal species.

There were also safety considerations in the specific design of our project. For example, though bacteria is directly involved in the protein production process, the final consumer product has minimal risk of contamination from bacterial components. First, the protein produced from the E. coli undergoes HIS tag purification before being incorporated into the emulsion system. This ensures that no bacterial components actually contact the oil or the chlorophyll. Moreover, even if minute bacterial contamination does occur, the water-in-oil emulsion in itself prevents any bacterial or protein components from making contact with the consumer product--the oil. The chlorophyll binding proteins never make contact with the oil since they are introduced into the oil in the form of water droplets, with surfactants further stabilizing the emulsion system. Overall, through these factors we have ensured that the final consumer product is safe for use by the public.

The application of pheophorbide as a fungicide was also carefully considered to ensure minimal impact to the general public and environment. Primarily, pheophorbide a is a natural catabolite of chlorophyll produced in plants. This means that although fungi are negatively impacted by exposure to pheophorbide a, the plants themselves are not affected due to inherent safety mechanisms against pheophorbide’s cytotoxic capabilities within them. Additionally, our application of pheophorbide a as a fungicide has minimal effect on humans. After irradiation with light, it loses its cytotoxic ability, making it an ideal fungicide safe for human use and consumption (REFERENCE). Due to these reasons, we believe pheophorbide’s risk is relatively low; however, we would need to confirm that it does not pose any threat to other animals and insects in the environment.

At the beginning of the project all team members, mentors, and Principal Investigators had to complete lab safety courses developed by the University of Calgary Environment Health and Safety (EHS) services to start work in the lab. These courses included occupational health and safety, laboratory safety, hazard assessment, incident reporting and investigation, spill response, biosafety, and WHMIS. By completing these courses, everyone on the team was equipped to work safely in the lab and were able to handle incidents or hazards appropriately if anything were to occur. Additionally, the University of Calgary Biosafety Committeeoutlines specific guidelines for following safe biological laboratory practices. Throughout the project, all team members observed and adhered to these guidelines during our experimental work.

ATCC - American Type Culture Collection. (2016). 293T (ATCC CRL-3216). Retrieved from https://www.atcc.org.