1. Would any project ideas raise safety issues in terms of ______?

A. Researcher Safety:

As with any science experiment, there are risks involved with our project. Our team works with bleach which can cause irritation and a burning sensation if improperly exposed through inhalation, ingestion, or prolonged exposure to skin. However, any person is able to buy bleach from their local grocery store. Even so, we take care to prevent any issues. Also, our team works with E. coli K12. It is a version of E. coli that is widely studied and found to be very safe and practical for research. In fact, it is one of the most-studied bacteria of all time. E. coli K12 has no known survival mechanisms in the environment and has a very low risk of causing harm to organisms. E. coli K12 is in attenuated strain that is approved for use in high school classrooms. Even with antibiotic resistance, the amount of harm caused to most organisms would be minimal. The only real danger would be if a bacterium was able to conjugate with another type of bacteria that was more harmful and transfer its resistances. The chances of this happening are small due to the fact that the bacterium is held in a lab that most high school students do not enter (separate room that remains locked) and because proper safety precautions are followed relative to our biosafety level when inside this lab, including proper safety equipment and diligent hand washing.

B. Public Safety:

There are not many reasonable threats to the safety and health of the public if the project were to be released. This is mainly because we are in the biosafety level (BSL) 1 category which limits what materials we can use. In saying this, bleach can be purchased by almost anyone at their local supermarket or grocery store. Therefore, the risk is no different than someone buying the bleach and using it as a household chemical. Also, E. coli K12 can be purchased by the public. Therefore, it is again low risk. As long as proper regulations are followed specific to their safety level, there is minimal risk. For the general public, the only way the bacterium may cause harm is through the spread of antibiotic resistance to chloramphenicol. The reason for this particular drug resistance is because this is what our team uses as a selective marker to attain the final product. It only becomes risky when this resistance is spread to more harmful bacteria.

C. Environmental Safety?

There are no unreasonable, adverse effects associated with bleach being introduced into the environment. E. coli K12 has no known way of surviving in the environment and cannot produce spores. It is also not able to colonize the gut of an organism which makes it a very low environmental threat. From our machine, there are no real environmental threats. Again, this arises from working within BSL 1 and from the public having access to the same materials we use. Therefore, they are all relatively stable and safe. There is no real toxicity other than bleach, but it can be purchased at many stores and locations around the world and is only harmful from intentional misuse (ex. ingesting very large quantities, inhaling large amounts). Our machine itself is not pathogenic but if it is able to conjugate with other possibly pathogenic bacteria, then it may cause some degree of harm. If the bacterium is isolated then this will not happen and may be used safely.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

The BioBrick parts that we built do not raise any major safety concerns. A few minor risks are associated such as the potential conjugation and sharing of antibiotic resistance. However, the largest safety issue for this year’s project is in the testing of our sensors. We have sensors that test for heavy metal contaminants. Hence, we need to use heavy metals to test the functionality and sensitivity of our detectors. We used Lead Acetate (0.1 M) and Cadmium Nitrate (1 M), both of which are available for purchase from a science supply store.

If yes,

A. Did you document these issues in the Registry?

We have not added parts to the registry at this point; proper documentation will be included when our BioBrick parts are added to the registry.

B. How did you manage to handle the safety issue?

Extra care was taken when working with these materials such as the use of a fume hood and proper lab technique. Since we are in a high school setting, we must abide by all BSL 1 guidelines and regulations. This severely limits our available options. Many regulations are imposed upon our team; a few include: a sink must be present in the lab, controlled access (locked doors), no eating or drinking in lab area, decontamination of surfaces after lab work, and a lab supervisor. All work is supervised by our instructor, Ms. Anne Byford; she has a background in molecular genetics and trains all team members to ensure safety. She has also supervised and gone through the specific methods and processes we use to isolate, cut, ligate, and view the DNA used to create our bacterium. This also includes proper lab etiquette and how to use equipment such as a microcentrifuge, pipettes, and gel electrophoresis boxes. Proper safety technique is used when handling all materials. We treat everything as if it were hazardous.

C. How could other teams learn from your experience?

Other teams could potentially learn from our experience through proper research on their biosafety level and its respective regulations. Also, teams could look at the materials used to test their machines, not just the construction process itself.

3. Is there a local biosafety group, committee, or review board at your institution?

A. If “yes,” what does your local biosafety group think about your project?

At Gaston Day School we do not have a set of our own biosafety rules. In saying this, we do follow all of the regulations and rules from BSL 1 and operate under the supervision of our advisor, Ms. Anne Byford, who has much knowledge in the biological field.

All of our iGEM projects at Gaston Day School are approved by the Head of the School, Dr. Richard Rankin, the Head of Middle and Upper School, Mr. Greg Rainey, and lastly the Board of Directors to ensure the safety of our team and school.

B. If “no,” which specific biosafety rules or guidelines do you have to consider in your country?

The United States of America’s biosafety regulations are controlled by the CDC (Center for Disease Control and Prevention). More information can be found in the link below.

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices, and systems be made even safer through biosafety engineering?

A. Safety is the most important part of the competition. Safety must be stressed upon and needs to be a priority in all labs. To help with this, each team and advisor should have to fill out a survey that measures the safety in their lab and also makes sure that they are staying within the regulations of their BSL. Also, photos should be submitted of the lab to show that they have the proper safety regulation and requirements.

Citations:

1. http://epa.gov/biotech_rule/pubs/fra/fra004.htm

2. http://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf pages 30-33.