Team members were all required to complete a Workplace Hazardous Materials Information System (WHMIS) safety course before they entered and worked in the lab. All BSL-1 precautions were followed when completing any lab work. The University of Lethbridge provides access to SynBridge on campus. A state-of-the-art synthetic biology maker space.

While the protein biosensor will be expressed using E. coli BL21(DE3) and long term storage of the plasmid will be in E. coli DH5alpha, we hope to eventually express the protein in denitrifying bacteria. Previous iGEM teams have demonstrated that the denitrifying bacteria Paracoccus denitrificans is easy to genetically manipulate and most common molecular techniques are applicable (Virginia 2017). This is a BSL-1 species and non-pathogenic. All standard safety procedures will be followed when using it in the lab.

Since we would eventually like to see the protein biosensor incorporated into the denitrification pathway of aquaculture systems, preventing the escape of any genetically engineered bacteria from the filter system is very important.

Physical containment: Using a physical container that has proper filters in place would be a requirement. Partnering with a company such as Aquatics Solutions to use their biological detector would be a good start.

Biological containment: We could engineer our bacteria to be dependent on a specific nutrient. P denitrificans have been shown to have growth defects in low cobalamin (B12) conditions [1]. This natural deficiency could be utilized to control and maintain growth of our engineered bacteria.

Waste Management: When a farmer or pet owner needs to fix or change their aquatic system, it may be necessary to remove the filter with the denitrifying bacteria. We would provide them with specific sterilization techniques and supplies.

SynBridge Lab Space.

Biosensors are generally developed for beneficial purposes, like any technology, they could potentially be misused. Malicious actors might use biosensors to detect and harvest dangerous pathogens from the environment, which could then potentially be used for bioterrorism. The biosensor could be manipulated or falsified to give incorrect readings, potentially leading to false alarms or misdirection of resources in a public health context. Information obtained through biosensors could be misused to discriminate against individuals or groups based on their biological characteristics. Therefore, implementing strong regulations and oversight for the use and distribution of biosensor technology is necessary.