Team:Nagahama/Safety

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Safety Form

Your Training

a)Have your team members received any safety training yet?

Yes, we have already received safety training.

b) Please briefly describe the topics that you learned about (or will learn about) in your safety training.

Our country ratify the law concerning the conservation and sustainable use of biological diversity through regulations on the use of living modified organisms. So we have learned the law of recombinant DNA experiment in Japan, the difference of experimental conditions required for various biological species, the method for sterilization of living modified organisms, and the rule for starting a new recombinant DNA experiment in our institute.

c) Please give a link to the laboratory safety training requirements of your institution (college, university, community lab, etc). Or, if you cannot give a link, briefly describe the requirements.

Sorry, our institute doesn't open the laboratory safety training requirements to the public. In our institute, reseachers and students who are planning to start a new recombinant DNA experiment, must receive a lecture about recombinant DNA experiment that are held by our genetic modification safety committee before starting the new recombinant DNA experiment.

Your Local Rules and Regulations

a) Who is responsible for biological safety at your institution? (You might have an Institutional Biosafety Committee, an Office of Environmental Health and Safety, a single Biosafety Officer, or some other arrangement.) Have you discussed your project with them? Describe any concerns they raised, and any changes you made in your project based on your discussion.

We have a genetic modification safety committee in our intitute, and our team primary instructor is a member of the committee. Our project have been discussed and permitted by the genetic modification safety committee. There was no concern and change during the discussion.

b) What are the biosafety guidelines of your institution? Please give a link to these guidelines, or briefly describe them if you cannot give a link.

Sorry, our institute doesn't open the biosafety guidelines to the public. Brieafly, reseachers and students who are planning to start a new recombinant DNA experiment, must receive a lecture about recombinant DNA experiment that are held by our genetic modification safety committee, and permission of new experiment before starting the new recombinant DNA experiment. In our institute, only P1, P1A, P1P and P2 experiments can be permitted.

c) In your country, what are the regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link.
Biosafety in Japan

http://law.e-gov.go.jp/htmldata/H15/H15HO097.html (Japanese version)

http://eiyaku.hounavi.jp/eigo/h15a09701.php (unofficial English version)

MEXT LifeScience Portalsite

http://www.lifescience.mext.go.jp/bioethics/index.html (Japanese version)

http://www.lifescience.mext.go.jp/english/index.html (English version)

The Organisms and Parts that You Use

File:Nagahama Safety2015 Spreadsheet.xls

Risks of Your Project Now

Please describe risks of working with the biological materials (cells, organisms, DNA, etc.) that you are using in your project. If you are taking any safety precautions (even basic ones, like rubber gloves), that is because your work has some risks, however small. Therefore, please discuss possible risks and what you have done (or might do) to minimize them, instead of simply saying that there are no risks at all.

a) Risks to the safety and health of team members, or other people working in the lab:

The marA activating AcrAB-TolC efflux pump, also efflux antibiotics from engineered E. coli, resulting more resistance of the engineered E. coli to antibiotics. Because the host E. coli has no toxic to human, there may be no risk to human safety. All engineered E. coli have been sterilized by autoclave sterilization. We use hexane to dilute geraniol and farnesol and use decane to collect antibacterial fragrances from E. coli culture. Hexane and decane are flammable. These solvents are not used beside the fire. We have worn rubber gloves and have sterilized all wastes to reduce the risks, including safety level 1 procedures.

b) Risks to the safety and health of the general public (if any biological materials escaped from your lab):

We only use E. coli JM109 and E. coli DH5alpha encoding extra DNA parts that are originated from E. coli and in Spring 2015 Distribution. These organisms are derived from E. coli K-12 strain that belong to Risk group 1. So we think that these organisms have no risks to the safety and health of the general public.

c) Risks to the environment (from waste disposal, or from materials escaping from your lab):

We are using only E. coli JM109 and E. coli DH5alpha. They belong to B1 level in biological safety in Japan, and can not live in nature. So if they escape from our lab, there is no risk to the environment.

d) Risks to security through malicious mis-use by individuals, groups, or countries:

All E. coli and DNA parts are stored in private refrigerators for our project, and the refrigerators have been locked when we do not use them.

e) What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)

As described above, we only use B1 level E. coli in our project and all biological materials are autoclaved before waste according to our institute rule.

Risks of Your Project in the Future

What would happen if all your dreams came true, and your project grew from a small lab study into a commercial/industrial/medical product that was used by many people? We invite you to speculate broadly and discuss possibilities, rather than providing definite answers. Even if the product is "safe", please discuss possible risks and how they could be addressed, rather than simply saying that there are no risks at all.

Q) What new risks might arise from your project's growth? (Consider the categories of risk listed in parts a-d of the previous question: lab workers, the general public, the environment, and malicious mis-uses.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks? (For example: auxotrophic chassis, physical containment, etc.) Such features are not required for an iGEM project, but many teams choose to explore them.

In our project, we have used E. coli JM109 strain, a derivative of E. coli K-12 strain encoding auxotrophy, which cannot grow in natural field. In future use, other organisms that produce more antibacterial fragrances naturally and have more tolerance to the fragrances, will be engineered to produce more antibacterial fragrances. The engineered organisms will be used under supplication of required nutrition, so they cannot grow outside the Flavorater. The Flavorater have the risk that the engineered organisms grow in food. We must need to separate them from food area by packing or separating with membrane filters that the engineered organisms cannot pass through. Only fragrances should be dispersed into food area. This approach will decrease the risk of food contamination by engineered organisms. We have introduced the marA into E. coli to make the host more tolerant to antibacterial fragrances. The marA also enhanced the engineered E. coli more resistant to antibiotics as a side-effect. When we use other organisms that originally have more tolerance to the antibacterial fragrances, the marA need not be transformed to the engineered organisms, resulting no change of resistance against antibiotics.