Team:SCAU-China/Safety

1. 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.

Members in our team have read the established laboratory safety principles of our school and the topics in our safety training can be summarized as follows:

I.Non-biological operations:
　　I.1 Handling toxic chemicals
　　I.2 Emergency measures(such as how to tackle fire, electric leakage and negligent wounds)
　　I.3 Instruments and facilities operation principles


II Biological operations:
　　II.1 Potential threats of our engineering bacterium (Escherichia coli)
　　II.2 Effective protection during organism operations
　　II.3 Waste materials handling measures
　　II.4 Emission rules

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.

The laboratory safety training requirements of our institution can be summarized as follows:

1.Trainees should comply with laboratory rules and should not precede any operations without permission.
2.Trainees should follow the trainer’s guidance and take notes during the training.
3.During demonstration of biological operations, both trainers and trainees are required to wear lab coat and other protective equipment.
4.Trainer should lay emphasis on safety issues of biological operations related to genetic modification and microorganism emission and announce the relevant rules.
5.Trainees cannot precede any experiment by themselves until the end of the training with a final operation examination

2. 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 contacted with a professor who is responsible for biosafety.

What the concern about are describe as follows:
I. Q:We undergraduate students are not quite familiar with the laboratory protocols and experience which may cause safety problems (such as infection, poisoning etc.) while doing our project.
A: We have graduate students as our supervisors to ensure our operation correctness preventing safety problems. All of our team members have at least been working in a lab for 3 months and received biosafety training.

II. Q:How risky are our genetic modified organisms, while exposed to the environment, to human health or experiment.
A: Materials that we use in the project are low risk engineering bacteria (E.coli K12 MG1655). And we have now set up the rules in our project to handle wasted materials before emission to minimize the risk to the environment and human

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.

The biosafety guidelines of our institution can be describe as follows:

1.All laboratories must be specially design and should set up strict management systems, standard operation procedures and rules.
2.Each staff should be equipped with personal safety equipment to avoid direct contact with the pathogenic microorganism or toxic chemicals.
3.New staffs should be well trained and should pass the experiment test before performing experiments themselves.
4.One person at least in each laboratory should take charge of biosafety and establish a continuous biosafety training program.
5.Laboratories should establish emergency handling procedures and have routine inspection for all of the equipment.

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.

http://www.gov.cn/zwgk/2005-05/23/content_256.htm

4. Risks of 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:

Though the organisms we use are at low risk, they are infectious with the possibility to cause human disease such as diarrhea, festers even enteritis. We wear rubber gloves in each experiment even cleaning the flasks or test tube. We treat volatilized solvent in the fuming cupboard. We wear disposable gloves when we use agarose gels (containing EB) and SDS gels (containing acrylamide). Each device is exclusively used in the gel running area such as pipet, measuring cylinder ect. We wash our hand before leaving the lab or drinking and eating with hand sanitizer. Eating and drinking is strictly forbidden in the lab working area.

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

It is possible, even in small rate, for the antibiotic resistant gene that we transduce into the bacteria to transfer from our engineering cells to some other infectious microorganisms through horizontal gene transfer, which may cause public health problems. We treat each tube and petri-dish of bacteria with Dupont Virkon before we pull them into the bacterial waste tank, which will be sent to specific biological company for further treatment.

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

To some degree, the exposal of the organisms in our project may cause pollution (to the air or water source), but the cells we use will not secrete toxic materials into the environment. In the HPLC experiment volatilized organic solvent must be use. We open the windows and exhaust fan to ensure ventilation of our lab. We also collect the elution solvent in a bottle and handle to company for further treatment.

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

The organisms we use are not strongly infectious, which means it will be risky only when we have a long-term direct physical contact with them or intake a certain amount of these cells by mistake. Even though, trainer have emphasized no one is allow to take away any materials in the lab.

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

We are amenable to ensure biosafety in our project and we do have set up rules and emergency measures to avoid accidents. To minimize the risk of microorganisms to team members’ health we have established a set of operation rules that helps us to normatively operate in the project. For example, we are supposed to wear surgical masks, lab coats and gloves to prevent physical contact with the microorganisms. On the other hand, we have our standard procedures to handle the wasted materials before disposal, such as diluting and sterilizing, to minimize their influence to the environment and public.

5. Risks of our Project in the Future

a) 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 methodsyou develop became widely available?

Suppose that our project grows into industry use, we evaluate the potential risks as follows:
1.The infection possibility might increase if lab workers/stuffs stay in workshop for a long time, exposed to the microorganisms, in condition that the microorganism operations such as inoculations and sampling cannot be conducted by machines.
2.Tons of reaction solutions containing E.coli might be risky if negligence appears through the process of production and maintenance of equipments. Once leak of microorganism happens by mistake or negligence, surrounding water would be contaminated which might influence the quality of domestic water and be risky to ambient public health.
3.The leak of microorganism might also threaten the environment. Some secreta of microorganism might be toxic to some plants or animals and the antibiotics resistance gene might be transfer into some other infectious microorganisms in higher risk level, through genetic flow.
4.Malicious mis-uses of this project may occur in form of swindle using specific genetically modified organisms or devices stolen from this project to make profit.

b) 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.

We are planing to design some practical methods to minimize risks:
1.We are trying to construct a part that including lacZ gene promoter controlling the expression of cytotoxic protein gene ccdB, before letting out the waste, we can add a small amount of IPTG to induce the expression of CcdB protein resulting death of cells.
2.In our project, the main microorganism we use to work are E.coli strain whose arcA gene has been knock out. In anaerobic environment, once culture substance or energy is in low level/being exhausted, the cells themselves would collapse due to imbalance of fast metabolic rate and low level energy source. So before we dispose the waste, we would store them in a hermetic and anoxic place for several days until the nutrient solution exhaust and the cells die out. We also have standard diluting and sterilizing procedures before waste disposal.

6. Further Comments

Our wet lab work have just been started now. Currently we are constructing an expression vector and a gene-knock-out vector, both of them are for the aim of enhancing the metabolism strength of the engineering bacteria to improve of MFC's （Microbial Fuel Cell） electricity production density. All of our members are trained for safety operations and biosafety issues and we have discuss about setting up a series of laboratory operation rules to normalize the operating steps.