While our project aims to help alleviate environmental pollution, we must take precautions to ensure that no personal or environmental harm comes in the process. Biological lab work comes with an inherent risk, and outlined below are specific risks associated with our project as well as precautions we take to safely complete our project. Our completed safety form can be found here.

Specific Safety Concerns

Laboratory Safety: To visualize DNA gels, we regularly use ethidium bromide, a DNA intercalating agent known to be carcinogenic. To visualize gels, we also use a powerful UV light. We regularly use the antibiotics chloramphenicol and ampicillin, which can be harmful to humans in large doses. For our project this year we are using salts of the heavy metals nickel, lead, and mercury. These metals can be acutely toxic at high enough concentrations and carcinogenic over the long term. We also use open flame alcohol burners to maintain a sterile environment.

Environmental Safety: If any biological materials escape from the lab there is a risk of transfer of antibiotic resistance from our engineered strains into other organisms. None of the coding regions we're using in our project should provide competitive advantage in a natural environment (unless in an environment with high metal concentrations), so the biggest risk is from transfer of antibiotic resistance to outside organisms. Additionally, release of the heavy metals we are working with this year (Ni, Hg, Pb) could also pose environmental and personal harm if released into the water supply.

Chassis Organisms: The table below describes all strains worked with for our project this year. The chassis organisms E. coli DH5a and BL21-AI are biosafety level 1 organisms and pose no disease risk to people. None of the coding regions we're using in our project should provide competitive advantage in a natural environment (unless in an environment with high metal concentrations), so the biggest risk is from transfer of antibiotic resistance to outside organisms.

Species name	Risk Group	Risk Group Source	Disease risk to humans?	Part number/name	Natural function of part
Escherichia coli DH5α	1	NIH	No	N/A	N/A
Escherichia coli BL21-AI	1	NIH	No	T7 polymerase	RNA polymerase native to T7 bacteriophage
Escherichia coli BL21-AI	1	NIH	No	BBa_K1460001	GST enzyme transfers reduced form of glutathione to xenobiotic substrates. YMT binds to metals to confer metal tolerance.
Escherichia coli BL21-AI	1	NIH	No	BBa_K1460002	GST enzyme transfers reduced form of glutathione to xenobiotic substrates. YMT binds to metals to confer metal tolerance.
Escherichia coli DH5α	1	NIH	No	BBa_K1460003	NixA transport nickel ions in H. pylori
Escherichia coli DH5α	1	NIH	No	BBa_K1460004	merT/merP transport mercury ions in P. aeruginosa
Escherichia coli DH5α	1	NIH	No	BBa_K1460005	CBP4 is used in N. tabacum to confer nickel tolerance and transport lead
Escherichia coli BL21-AI	1	NIH	No	BBa_K1460006	Composite Part
Escherichia coli BL21-AI/td>	1	NIH	No	BBa_K1460007	Composite Part
Escherichia coli BL21-AI	1	NIH	No	BBa_K1460008	Composite Part

Safety Protocol

Wetlab: All lab members wear nitrile gloves, closed-toe shoes, and use eye protection when working with volatile chemicals or UV light. Gloves are replaced and hands are washed immediately after using ethidium bromide or any of the metal solutions. Members work in small groups to ensure if any harm comes to one, others are there to assist. When working with a new reagent or piece of equipment, a faculty lab manager or experienced member is always present to assist.

There are taped off, designated areas for working with both ethidium bromide and the heavy metal solutions. These areas are cleaned before and after work and are the only areas these solutions may touch. All toxic waste is placed in a specialized receptacle and is picked up and disposed of by Cornell Environmental Health and Safety.

All disposables that come in contact with biologics are disposed of in biohazard waste. The lab space also contains sharps containers for disposal of all sharps that contact biological material. All biohazard waste is autoclaved and transported to the building's centralized waste facility where it is disposed of as regulated biological waste.

We maintain 2 copies of MSDS's for every chemical we use in the lab: one for our own records and one for the lab manager and users of the lab space who are not part of our team (list of all chemicals is attached as an excel document). The lab is equipped with flame-retardant benches, spill kits, safety showers, eye-washes, and fire extinguishers.

Dry lab: We use the Emerson Machine Shop for fabrication; each of the dry lab subteam members has attended the prescribed training session for use of the shop and has learned to use each of the tools safely. Each member of the dry lab subteam was trained in the safe usage of the milling machine and the metal lathe.

All machine shop work is conducted under the supervision of the Emerson machine shop staff. Safety goggles were worn at all times. Masks and gloves are worn as appropriate. Closed-toe shoes and long pants were also worn when working in the machine shop. While working in the machine shop we maintained a clean work environment so we could maintain visibility at all times. When lifting heavy objects, proper lifting technique was used, and an appropriate number of individuals were used for lifting said objects.

Training and Enforcement:

Training: All team members who work in the wet lab must complete Cornell EH&S general lab safety and chemical waste disposal courses prior to the onset of work. These courses set specific guidelines and are the standard requirement for work in a biosafety-level 1 lab at Cornell. Additionally, all team members must complete a lab orientation session with the manager of the BME instructional lab, Dr. Shivaun Archer. During these sessions, Dr. Archer familiarizes new members with the safety equipment and procedures specific to the labspace in which we work.

Prior to the onset of work for the year, all new members are required to go through a 1 week training program. During this program, safety officers reinforce safety procedures learned during the EH&S courses, discuss safety protocol pertaining to specific chemicals with which we work, and ensure all lab members fully understand all safety procedures.

Safety Officers: The safety officers were chosen to be team members who could directly supervise the activities of the other team members. One team member each was chosen for the wet and dry lab subteams to ensure that all team members are working safely, whether with bacterial cultures or power tools. These team members also act as liaisons to the wet lab and machine shop managers and, when necessary, the Institutional Biosafety Committee to ensure proper equipment usage.
These team members are responsible for discussing the proposed workplan for the project with the wet lab and machine shop managers before starting work to ensure that it is safe to continue. In the case of the wet lab in particular, this involves going through a detailed list of protocols, including all organisms, chemicals, and genetic constructs being worked with, to ensure conformity with the Environmental Health & Safety guidelines. They must go through the same safety training as all other team members, but are required to redo the training each time we recruit new members in order to keep up-to-date with safety considerations. In addition, they maintain contact with the supervisors of the workspaces, usually in the form of a weekly check-in, to discuss any safety concerns that have arisen and ensure that equipment continues to be used properly.

Enforcement: Team members who violate safety rules are required to work under supervision of the safety officers for the remainder of the week, or until the safety officer believes the member is capable of perform the task unsupervised. For multiple infractions or complete disregard to safety protocols, a member may be restricted from laboratory work until he/she undergoes EHS chemical safety online training again, and demonstrates proper performance to a team leader of failed technique(s) in a controlled setting.