Basical questions about our 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.

During the safety training, we were instructed on the following topics.
1. Methods of Protection and Rules for Conduct when working in the laboratory.
2. Transport, Disposal and Disinfection of biological wastes, ethidium bromide gels, broken glass wares, etc.
3. In case of danger during fire or injury.
4. Operating instructions of laboratory instruments and equipments.
5. Safety hazards concerning various chemicals.
6. Common Do’s and Don’ts.
7. How to write proper records of the work in the laboratory to keep track of the progress and to troubleshoot errors.

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.

http://www.uni-goettingen.de/en/401.html
(This is a website for the safety of genetic engineering from our university.)

About our 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.

A subgroup for genetic engineering under the department for security and environmental safety is responsible for the biological safety at our university. Every staff member in the Georg-August University Göttingen is entitled to give instructions to colleagues and/or students regarding content and structure of the work, as far as Health and Safety Regulations are concerned. We have discussed our project in detail with our instructors and they are well aware of the safety concerns that might arise in our lab and at the same time, have apprised us of the same. Weekly meetings with our instructors helps us to avoid/overcome any issues that may have arisen.

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.

http://www.uni-goettingen.de/en/100640.html
(This is a link of the information about the Safety/Environmental Protection Section.) from our university.

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.

In Germany, the Federal Office of Consumer Protection and Food Safety put forth the guidelines for biosafety in research laboratories. The following link gives detailed description about the safety regulations in Germany
(www.bvl.bund.de/EN/06_Genetic_Engineering/genetic_engineering_node.html).

The Organisms and Parts that We Use

HERE is the excel form of our organisms.

Risks of Our Project Now

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

We are currently working with cDNA/RNA of S1 and S2 level organisms cloned in E.coli DH5alpha and expressed in S.cerevisae. Although these organisms are not pathogenic to humans, lab coat and gloves are worn at all times during work for protection. Furthermore, the work with such organisms is restricted to sterile laminar air flow chambers. We are using ethidium bromide, an intercalating agent, for staining DNA in agarose gels. We are aware of the fact that ethidium bromide is a mutagen and carcinogen. As such, the work with ethidium bromide is only permitted at a marked area in the laboratory and it is essential to wear nitrile gloves. The agarose gels containing ethidium bromide are disposed of in a specially marked container. While working with ethidium bromide gels, it is necessary to wear a lab coat, nitrile gloves and a face protection to avoid UV damage. Furthermore, a risk while working with agarose is boiling retardation. In order to minimize this risk, every student is required to wear special heat-stable and thick gloves. Since Antibiotics encourage the emergence of resistant microorganisms (Bacteria and Fungi), adequate precautions were taken to minimize the chances of the same. The concentrations of the antibiotics used in this project are relatively low and steps were taken to minimize exposure; to both the skin as well as general work environment. As it is necessary to guarantee emergency service; therefore there are always two or more students present in the laboratory for the same. Additionally, during the S1 security briefing, we were introduced to the emergency eye shower system and shown the emergency exits. The risk of a fire through the Bunsen burner is very low. In case of a fire, an alarm signal would start immediately.

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

The chances of any biological material to escape from the laboratory are none, because all biological materials are disposed of separately after proper inactivation. The strain of E. coli used DH5alpha was developed in the laboratory, for laboratory cloning procedures.

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

The biological wastes disposed, do not pose any threat to the environment, since they are autoclaved as per instructions and disposed in suitable containers. Moreover, the bacterial strain we use (E.coli DH5alpha) for genetic engineering applications like cloning was developed in the laboratory, for laboratory cloning procedures, and therefore has no 'natural' environment or habitat as it does not exist in nature. It is also incapable of transferring its genetic material to other bacteria in case if it is accidentally released into the environment.

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

We are currently working with cDNA/RNA of S1 and S2 level organisms cloned in E.coli DH5alpha and protein parts expressed exogenously in S.cerevisae. These organisms or the protein parts are not a threat to the environment or to humans. In infinitesimal chance of malicious misuse by individuals, groups or countries, the organisms / parts do not pose any security risk as they are not in anyways toxic/pathogenic to humans or the environment.

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

Safe lab practices and strict safety measures are followed in the lab. All biological materials are disposed of separately in special waste containers after proper inactivation by sterilization.

About risks of our project in the future

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?

In the future, in vivo studies is crucial for designing a therapeutic product. During such studies we may have to work with pathogens belonging to Risk Group level 2, so proper care and precautions have to be taken beforehand. All such experiments have to be carried out in suitable work area designed as per set rules.

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.

Currently, our project does not include any design features to reduce risks, as it does not pose any threat / risk to humans or the environment.
As a future perspective, our technique can be developed as a therapeutic tool to fight against fungal pathogens. Even then the peptides we use for such purposes does not have any ill effects / risks that would evoke safety concerns.
But care and caution must be taken during in vivo studies with potentially harmful pathogens.

The Basic safety form in the lab(Due on June 23rd)

1. a) Does your country use a four-part "Safety Level" rating system for laboratories? (The system might be called in English "Risk Levels", "Bio-Safety Levels", "Containment Levels", "Bio-Security Levels", or some similar name.)
If yes, which level is used for the most dangerous organisms?

• Yes. Level 4 is used for the most dangerous organisms. (True for most countries in Asia, the European Union, and North/South America. This is equivalent to the WHO system.)

b) What is the Safety Level of your lab? (Use the WHO numbering system, in which Level 4 is used for the most dangerous organisms).

• Level 1 (low risk, ~= WHO BSL 1)

2. a) What type of work environments do you use to handle biological materials? Please check all that apply.

• Open benches
• Laminar flow hood / biosafety cabinet with open front

b) Do you handle different materials in different work environments? If yes, please describe what materials you handle in what work environments.

Yes.
1. Open Benches: For Molecular Biology Techniques such as PCR, Gel Electrophoresis, Gene Cloning.
2. Laminar flow hood: Inoculation, Subculture, Plating, Picking colonies from plates.
3. Fume hood: When handling concentrated acids and other volatile substances.
4. Marked Area: Gels are treated with ethidium bromide (EtBr) in such marked work areas and Nitrile gloves are worn at all times, when working in those environments. The agarose gels containing ethidium bromide are disposed of in a specially marked container.

3. a) What personal protective equipment do you use in your lab? Please check all that apply.

• Lab coats
• Gloves
• Safety glasses / goggles

b) Do you use different protective equipment for different procedures? If yes, please describe what equipment you use for what purposes.

Yes. 1. Nitrile gloves: When working in marked EtBr area.
2. Goggles and Safety Glasses: Under Fume hood.
3. Gloves: When dealing with molecular biology techniques.
4. Lab coat: Generally at all times.

4. How do you dispose of biological waste? (For example: liquid cell cultures, agar plates, used pipette tips.)

We have separate disposal area for S1 wastes such as agar plates, used pipette tips, falcon tubes, eppendorfs, etc..
Liquid cell cultures are first autoclaved to remove live microbes and then are disposed of in S1 liquid waste containers.