Safety

Basic Safety Questions

1. Your Training
1. Have your team members received any safety training yet?

Yes, we have already received safety training.

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

We learned about recombinant DNA technology guideline and things to be careful of when working in laboratory. And one of team members, Kenta Nomura, had a training to learn handling of hazardous materials, and he teaches us tips for experiment.

3. 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.

For detail information about safety training please refer to the site below (Japanese): http://web.jim.gifu-u.ac.jp/sienka/DNA/

2. Your Local Rules and Regulations
1. 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.

The supervisor at our institution is Professor Hiroyuki Koyama, responsible for biological safety at Gifu University. A team instructor, Akio Ebihara, had already explained to him what we were going to do in our iGEM project. He mentioned that there is no special concern in our project because our experiment is similar to a practice course for undergraduate student. And Prof. Koyama permitted us to do the project.

2. 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.

Our University is under ISO14001. We follow its rules. Law concerning the conservation and sustainable use of biological diversity through regulations on the use of living modified organisms (Cartagena Agreement): http://www.gifu-u.ac.jp/images/12/rule/05dna.pdf

3. 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.

http://www.lifescience.mext.go.jp/bioethics/anzen.html http://www.bch.biodic.go.jp/english/cartagena/images/e_cartagena.pdf

3. The Organisms and Parts that You Use

Species name(including strain)	Risk Group	Risk Group Source	Disease risk to humans?	Part number/name	Natural function of part	How did you acquire it?	How will you use it?
Escherichia coli(K12)	1	ABSA	yes			Instructor	building our parts
Oryza sativa	1		no	[BBa_K118023]/cenA	involved in making and decomposing sugar chain	from iGEM HQ	decompose cellulose into cellobiose
Oryza sativa	1		no	[BBa_K118022]/cex	involved in making and decomposing sugar chain	from iGEM HQ	decompose cellulose into cellobiose
Oryza sativa	1		no	[BBa_K118028]/bglx	involved in making and decomposing sugar chain	from iGEM HQ	decompose cellulose into D-glucose
T4 phage	1		no			NBRC20004 Escherichia coli phage T4	building our parts

4. 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.

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

Since our experimental room is small, we sometime work in a crowded situations. So we need to be careful not to make a glancing hit. This might happen unexpected accidents. We use a cancer-causing reagent, ethidium bromide, and we should be careful to use it.

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

Biological materials or these products might be toxic if they escaped from our lab. Perhaps they escape by an earthquake, a fire, and so on.

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

Escaping bacteria from lab by disposing wastes without autoclave might harm the environment that includes natural life.

4. Risks to security through malicious mis-use by individuals, groups, or countries:

An entry to our lab space without any permission, which may occur after losing the key to our lab. The entry allows the individuals to do an abuse of the regents.

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

Follow the rule for lab safety. No entry for those who have little concern.

5. 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.

1. 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?

A toxic protein can be produced using parts we are going to make. Any materials in our lab should not be transferred without a permission from our instructor.

2. 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.

Be careful not to talk information and details of our experiment to others.

6. Other

In our project, we examined the use of Cadmium (Cd) and Zinc (Zn) to confirm whether the lengthened metallothionein combined with heavy metals. To use their metals safely, we discussed the potential dangers and how to treat them, with our team members and some professors.

First, Cd is harmful to living things, so one of our members anticipated giving damage to health of experimenters, causing pollution, and destroying ecosystem. But the damage to health of experimenters will not happen unless Cd enters the body through our mouths, and the pollution and the destruction of ecosystem do not happen when waste liquors are disposed following the standard rules, so there are no possible happening these problems.

Second, the lack of Zn causes the taste disorder, but on the other hand, excessive quantity of Zn is needed to cause symptoms of poisoning. So we considered that the dangers from Zn had almost nothing.




Now, experiments are conducted in consideration of safety, but the pollution from chemical materials may happen in the following cases:

In the case that…

• the polyethylene tanks to put the waste liquors into crack
• waste liquors attach to tubes, tips, and the water to wash with something
• disasters like earthquake happen (especially in Japan)
• someone who do not know about our experiment touches chemical reagents in the same laboratory
• desks or doors are touched unconsciously by the hands which have touched danger materials
• the dangers or characteristics about chemical reagents such as the contents of kits are not understood



To reduce the dangers in these cases, the following things are conducted in practice:

• the polyethylene tanks to put the waste liquors into crack
  →To prevent the waste liquors from flowing out to balcony or overflowing due to the rainwater, we set the trays under the tanks and store in covered containers.
• waste liquors attach to tubes, tips, and the water to wash with something
  →The resources polluted from waste liquors are collected in the prescribed position.
• disasters like earthquake happen (especially in Japan)
  →If rivers flood, the first floor can be submerged under the water, so laboratories are usually equipped on the first floor. If disposal facilities are destroyed, there is no help for it.



Next, to reduce the dangers in these case, the following things are considered to be effective:

• someone who do not know about our experiment touches chemical reagents in the same laboratory
  →All who use the same laboratory should understand the materials used in mutual works and the dangers of them.
• desks or doors are touched unconsciously by the hands which have touched danger materials
  →We should notice that we use dangerous materials.
• the dangers or characteristics about chemical reagents such as the contents of kits are not understood
  →To prevent dangerous chemical reagents from being disposed without being noticed that it is danger, we should make sure of the contents of the chemical reagents in the kits precisely. Users should understand the characteristics of their materials (for example, it is possible that toxic gas will be generated due to the release of weak acids) fully, and, use or dispose them on the premise that we understand their dangers.



Finally, we could not find how to resolve in the following cases:



• disasters like earthquake happen (especially in Japan)
  →If disposal facilities are destroyed, it is possible that toxic materials will flow out. The methods to prevent damages have still being looked for.
  →We should always pay attention to the strength of disposal facilities and the inspection of them, and renew them. And we should assume the unexpected things.