Team:Cornell/project/safety

Specific Safety Concerns

We must be concerned with the safety implications that our project could subsequently have on the natural environment, as we are modifying the genomes of organisms to produce novel functions. We have designed and are implementing a number of biological safety mechanisms to prevent the spread of potentially harmful genetic material to the environment. This includes a recombination system to remove and degrade engineered portions of fungal genomes before using the fungus for packaging production, as well as a multi-faceted killswitch to induce our organism to halt its own growth upon the addition of a very specific chemical cue. We have corresponded with our corporate collaborators to determine the sorts of precautions that may be necessary for commercial deployment and are working to incorporate them.

Laboratory Safety

Our project involves regular use of ethidium bromide, a DNA-intercalating agent known to cause cancer, as well as the use of powerful UV light, for visualization of gel electrophoresis. We must prepare culture media with antibiotics, which could be harmful to humans in large doses. We also work with ethanol lamps to maintain a sterile environment, which do involve having an open flame on the benchtop.

Chassis Organisms

The wet lab subteam regularly works with a non-pathogenic strain of E. coli; as part of our project this year, we are also working with two fungal species, Ganoderma lucidum and Cochliobolus heterostrophus, as well as Agrobacterium tumefaciens and Aspergillus niger, to a limited degree. These are all Biosafety Level 1 organisms and are therefore safe to work with within our existing lab space; however, these all require special precautions. Fungi, for instance, can release spores that can be irritating to eyes, skin and lungs, and damaging if inhaled or ingested in significant quantities.

Novel Coding Regions

None of our novel coding regions pose a threat to researcher safety; however, genes such as those for chitinase and antifungal selection markers could be harmful if released into the environment. We must therefore use care to ensure that our engineered organisms do not escape the confines of our lab space.

Safety Protocol

Wet Lab

In the lab all members wear nitrile gloves, closed-toed shoes, and use eye protection when working with volatile chemicals or using UV light. Members typically work in groups of 2-3 so as to be aware of each other’s safety and the work at hand. If using a new piece of equipment, a team member must be supervised by either the faculty lab manager or, in her absence, an experienced team member.

There are taped-off areas for work with carcinogenic substances (particularly ethidium bromide) as well as fungal specimens. When working with fungi that could potentially release harmful spores, team members must work in a designated area and wear masks and eye protection. The delineated and surrounding areas are cleaned with ethanol and/or bleach prior to and immediately following each use, and if these substances must come into contact with a non-designated area, it is cleaned thoroughly to ensure that no residue remains.

All disposables that come into contact with biological materials, including but not limited to: pipet tips, syringes, filters, membranes, Petri dishes, microcentrifuge tubes, culture tubes, and gloves, are disposed of in the biohazard waste. There are also specialized waste containers for glass and sharps accessible in the lab space. Though we are not working with any highly toxic substances this year, we have protocols in place for using and collecting waste from hazardous chemical and/or biological substances in a manner approved by Cornell EHS.

We maintain two copies of MSDS sheets for every chemical stock that we maintain in our lab; one for our own reference and the other for the lab manager and users of the lab space who are not team members. There is a chemicals hood available for use with any chemicals that may produce harmful fumes, i.e. acids and organics. The lab is also equipped with flame-retardant benches, spill kits, safety showers, eyewashes, fire extinguishers, and clearly marked emergency exits.

Dry Lab

We use the Emerson Machine Shop for fabrication; each of the drylab 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 drylab 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

Required Safety Training

All team members are required to take the EHS Chemical Safety and Laboratory Safety Training, as well as an orientation session with the building coordinator. In addition, before gaining access to the lab, all team members go through an orientation with our lab's manager detailing proper equipment usage and safety within our lab space. Before any independent lab work begins, wet lab members spend the first week of every summer project in a laboratory Boot Camp hosted by experienced team members and, if needed, graduate advisers. These training sessions ensure proper laboratory technique, protocol, attire, and safety. Dry lab members are required to undergo machine shop training in order to gain access to the Emerson machine shop, to similarly ensure proper equipment usage.

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.

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.