FATIH UNIVERSITY MEDICAL SCHOOL ANSWERS TO NEW SAFETY AND SECURITY QUESTIONS

1. Would the materials used in your project and/or your final product pose:
a. Risks to the safety and health of team members or others in the lab?

Actually few hazardous chemicals and solutions are used in some lab procedures such as gel preparation and electrophoresis. However these chemicals and solutions are used according to the safety rules of the laboratory with care and caution. All the members were trained for safety regulations of the laboratory as well as toxicity of the chemicals and solutions before starting the current project.

b. Risks to the safety and health of the general public if released by design or accident?

When released by accident, our parts and materials actually cause no negative damage to the general public. B.subtilis is known not to be associated with disease process in humans in regular conditions and thus are supposed to be benign by nature and present no danger to anyone. Only under stress conditions, B. subtilis may form spores and these spores are resistant versions of single cells. While B. subtilis itself isn't pathogenic, its spores may be infectious if inhaled. But all of our team members are trained for safety of the laboratory rules and use mask and gloves when dealing with bacteria. Besides all experiments are performed in synthetic laboratory designated by Fatih University Medical School and no material is carried out of the laboratory. However to increase public safety of our project, in case of accident, our plan is to destroy particles including B.subtilis by cleaning the biofilm from the spores and treating with Fenton reagent containing dissolved oxygen and other constituents.

c. Risks to environmental quality if released by design or accident?

Bacillus subtilis strain 3610 is a wild type strain that is already widespread in the environment. B. subtilis strain 168 is a derivative strain of the 3610 and has disabling auxotrophs mutations that makes it even less likely to colonize or cause harm to human or animal health. Our experiment does not include any modifications, which enhance their ability to survive or disseminate. Thus, there is no specific environmental risk associated with use of the Bacillus subtilis strains except its spore formation. In case of accident, cleaning and treating the spores with Fenton reagent as mentioned above provide environmental safety. Besides all procedures are performed in a specified laboratory.

E. Coli strains TOP10 and BL1 have very limited ability to survive outside the laboratory so that it would be unable to survive or disseminate. Therefore, there is no specific environmental risk associated with the E. coli strains. All bacterial waste are kept in 10% bleaching solution for one day, then are autoclaved to be sterilized.

d. Risks to security through malicious misuse by individuals, groups or states?

Please explain your responses (whether yes or no) to these questions. Specifically, are any parts or devices in your project associated with (or known to cause):

• pathogenicity, infectivity, or toxicity? No
• threats to environmental quality? No
• security concerns? No

As explained below:
Nothing could be used maliciously in our project. The materials we use are commonly used in experimental procedures in various laboratories all over the world and all are harmless and do not pose any threat to environment and public as well as security concerns.

Throughout the project, we used the Escherichia coli strains Top10 and BL1, Bacillus subtilis strains 3610 and 168. Wild-type E. coli is classified as a hazard group 2 pathogen by the UK Advisory Committee on the Dangerous Pathogens (ACDP) and Wild-type Bacillus subtilis (i.e. strain 3610) is classified as hazard group 1 organism by the ACDP and its derivative B. subtilis strain 168 has disabling auxotrophs mutations (e.g. conferring a requirement for tryptophan, Zeigler DR et al, 2008. J of Bacteriology) that makes it even less likely to colonize or cause harm to human or animal health. B.subtilis is at level 1 biosafety according to World Health Organization (WHO).

2. If your response to any of the questions above is yes:
a. Explain how you addressed these issues in project design and while conducting laboratory work.
b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry
3. Under what biosafety provisions will / do you operate?
a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible

Yes, our institution has its own biosafety rules. Rules for laboratory use, general principles, prevention from hazardous materials and application of emergency intervention in case of accident are included. The web link is given as (http://www.fatihmed.edu.tr/icerik/guvenlikkilavuzu.php).

b. 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

Yes. In Fatih University Medical School, Laboratory and Patient-Employee Safety Committee is responsible for control as well as biosafety of laboratories and safety of patients and employees. This committee works under one of the vice medical director of Fatih University Hospital, Prof. Dr. Mehmet Gunduz. Form on safety rules of Fatih University Medical School Laboratory use was filled in as required. We discussed our project with Prof. Dr. Mehmet Gunduz. Safety and security issues are found sufficient enough that no change is considered as necessary. (Further questions can be directed to Prof. Gunduz, tel: +90-312-203 5103, mgunduz@fatih.edu.tr).

c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.

Yes, we did. Our advisor provided us biosafety and lab training before starting of our project. In the training, general safety rules of laboratory use, prevention from hazardous chemicals and solutions as well as emergency intervention in case of accident were included.

d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.

Yes, Turkey has national biosafety regulations and the link is given as (http://www.tbbdm.gov.tr/Home/BioSafetyCouncilHome/BioSafetyCouncilHomeChoose.aspx).

4. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

First, IGEM committee may prepare a lab safety acknowledgement form like OSHA form. This form can be required to be filled in by all members. This will allow us to confirm whether members of the teams are informed about safety issues.

Second, a new prize related with security and safety issues may take place in the judging procedure separated from “Best Human Practices” prize. Existence of “Best Safety Part” or “Best Safety Application” can emphasize the importance of the safety issues and inspire iGEM teams to work on these issues and may give a chance to develop new ideas, to interrogate current safety and security applications in order to improve them.

Third, iGEM committee may organize a webinar to inform the participants about safety and security in labs. Webinar can be done at the beginning of the experiments. Mentor scientists may talk about their experiences and mention some tricks about safety issues.