Team:Yale/Safety

1. Would the materials used in your project and/or your final product pose:

• Risks to the safety and health of team members or others in the lab?
  
  Our laboratory is certified for Biosafety Level 1 work and Biosafety Level 2 work. Our work fell within the BSL-1 domain, as indicated per Center of Disease Control guidelines. Some materials are irritants, toxic upon inhalation, ingestion, or contact, are possible mutagens, or are flammable. Acrylamide is a neurotoxin. Sharps and broken glass can also pose threats to safety and health. UV light is also mutagenic. E. coli and C. elegans strains used in our experiments were not pathogenic. Goggles, gloves, and laboratory coats were worn at all times.
• Risks to the safety and health of the general public if released by design or accident?
  
  If toxic, flammable, or chemically reactive substances used in our experiments are released, they may post threat to the general public. These include physical hazards or health hazards. Hazardous decomposition products that decompose in water are a problem if they are carelessly flushed down the drain.
• Risks to environmental quality if released by design or accident?
  
  There are no additional risks posed by our projects compared to other general BL1 lab concerns. Our bacteria are not pathogenic and are unable to survive outside of the lab environment, because they are unable to effectively compete with other organisms in nature. They do not cause adverse reactions in immunocompetent humans. They do not cause infection.
• Risks to security through malicious misuse by individuals, groups or states?
  
  Antifreeze proteins do not generally pose risk to security, environmental quality, or safety. Many are already being used in industry and for consumer products. Lab materials present the potential dangers described above (flammable, combustible, causing irritation or burns, carcinogens, neurotoxins, etc). As such, there is possible risk to security through malicious misuse.

2. If your response to any of the questions above is yes:

• Explain how you addressed these issues in project design and while conducting laboratory work.
  
  Our laboratory is certified for Biosafety Level 1 work and Biosafety Level 2 work. Our work fell within the BSL-1 domain, as indicated per Center of Disease Control guidelines. All students working in the laboratory were required to complete the following set of training tutorials, including passing a test at the end.
  • http://www.yale.edu/ehs/onlinetraining/BiosafetyPart1/BiosafetyPart1.htm
  • http://www.yale.edu/ehs/onlinetraining/BiosafetyPart2/BiosafetyPart2.htm
  • http://info.med.yale.edu/chemsafe/
  • www.yale.edu/ehs/onlinetraining/hazwaste/chemicalwaste.htm
• All materials were used in accordance with local, national, and Yale's Biosafety requirements. Standard lab practices were followed, including secondary containment of chemicals, proper storage of volatiles and flammables, and separation of acids and bases. Nitrile gloves were worn at all times within the lab. Double nitrile gloving was used when handling ethidium bromide, a toxic chemical and suspected mutagen. A pipet was kept exclusively for ethidium bromide use. We obtained SYBR Safe to stain DNA in August, which is safer than ethidium bromide. Fume hoods were used when handling volatile compounds, concentrated acids and bases, and other reagents. Inhalation and skin contact was avoided. Chemical agents were properly disposed of in designated biohazard waste bins. When UV light was used to visualize gels or GFP, a UV-blocking shield was used. Absolutely no food was allowed in the lab.
• E. coli were decontaminated with a dilute clorox bleach solution. Anti-freeze proteins are non toxic, and are even found in some commercial ice-creams! All biological waste was stored in autoclave bags and was autoclaved prior to disposal. Sharps and broken glassware were disposed of according to institutional guidelines. Hazardous liquid waste was clearly labeled, and stored in secondary containment for disposal by the institution. Thus, although there is potential for harm to researchers, it is minimized through following procedures approved and used by many laboratories at Yale. It is also minimized by training and common sense.
• We do not anticipate threat to public safety. Organisms that we worked with are all non-pathogenic. They are likely unable to survive outside of the lab environment, because they will be unable to compete with other organisms in nature. Biomaterials were disinfected with chlorox and autoclaved after use. We did not use gloves to touch doors outside of the laboratory to avoid others coming into contact with our chemical and biological agents. There are no additional risks posed by our projects compared to other general BL1 lab concerns. The risks of environmental harm were mitigated by following the above protocols. Hands were washed before and after leaving the laboratory. Our new biobricks consist of novel and previously well characterized anti-freeze proteins. These are non-toxic proteins, and some antifreeze proteins are even used in food additives. Bacterial strains are non pathogenic.
• Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.
  
  This has and will continue to be done.

3. Under what biosafety provisions will / do you operate?

• Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible. The following rules of biosafety were followed during the course of our experiment: http://www.yale.edu/ehs/Documents/Bio/Biosafety%20Manual.pdf. It details protocols relevant to exposure incidents, emergency phone contacts, registration for the use of biological materials, human pathogens, recombinant DNA experiments, human gene therapy, human blood, body fluids, tissue, and other potentially infectious material, animals, biological safety cabinets and other laminar flow benches, training, medical surveillance program, accidents, risk assessment and management, routes of exposure, biosafety levels, signs and labels, wall signs, door signs, personal protective equipment, laboratory equiptment, decontamination and disposal methods, spill response, and shipping.
• Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review. Our project was overseen by the Yale Biological Safety Committee and the Office of Environmental Health and Safety (OEHS), as well as the Yale Animal Resource Center. Our project has been approved as consistent with Yale's safety regulations. No changes were needed as long as proper protocols were followed.
• Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training. See above. The following are relevant links: http://www.yale.edu/ehs/onlinetraining/BiosafetyPart1/BiosafetyPart1.htm http://www.yale.edu/ehs/onlinetraining/BiosafetyPart2/BiosafetyPart2.htm http://info.med.yale.edu/chemsafe/ www.yale.edu/ehs/onlinetraining/hazwaste/chemicalwaste.htm
• Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible. This is governed by the CDC (center for disease control and prevention) of the US Department of Health and Human Services: http://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf.

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?

All teams should be required to submit a letter of safety approval from relevant safety offices. Physical containment should not be the only safety measure taken by laboratories. One possibility is creating new organisms that proliferate only when exposed to a certain laboratory chemical. Because the lab chemical is not found in nature, proliferation outside of the lab is limited. Lab organisms might contain a "kill-gene", that is inactivated only when a laboratory reagent (rare in nature) is provided. All teams should also discuss and debate safety issues, and engage other students, policymakers, research adminsitrators, and commercial providers of raw materials for the research. Teams must continue to describe and document safety, security, health and/or environmental issues as they submit their parts to the Registry.