Safety

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- Safety Equipment
      - Waste Management
      - Recombinant DNA Technology and Biological Safety
   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.
      Laboratory Assistant, College students and Technician


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. Our institution has a committee dealing with environmental security, health, safety, and (K3L). But they did not concerned in biosafety of modified organisms. In addition, our biosafety committee (K3L) deals only in laboratory safety and environment security.
   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.che.itb.ac.id/safety
      http://www.tf.itb.ac.id/files/2011/11/SOP-K3-Laboratorium-Teknik-Fisika.pdf
      http://www.tf.itb.ac.id/files/2011/11/KEPUTUSAN-sop-kedaruratan.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.Yes, our country does. It can be accessed in http://indonesiabch.or.id/tentang-bkkhi/


3. The Organisms and Parts that You Use
   1. Species name (including strain)
      Escherichia coli DH5α and BL21(DE3)
   2. Risk Group
      Risk Group 1
   3. Risk Group Source
      NIH
   4. Disease risk to humans?
      E. coli may irritate the skin, eyes, respiratory tract, and blood
   5. Part number/name
      BBa_K936024; BBa_K936000
   6. Natural function of part
      BBa_K936024 = to convert ethylene glycol into glycolate; BBa_K936000 = hydrolyses cutin
   7. How did you acquire it?
      BBa_K936024 = requesting from HQ
   8. How will you use it?
      BBa_K936000 = to degrade PET into terephthalic acid and ethylene glycol; BBa_K936024 = to convert the product of PET degradation


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:
      Our topic is about whole cell biocatalyst that can degrade one kind of plastic, that is Polyethylene Terephthalate (PET). The whole process we strictly dispose the chemicals and toxic substances, such as Ethylene glycol. Ethylene glycol can be a mild irritant if it contact with the skin or if it is inhaled, so we have to protect our self by wears gloves, lab coats and always work with ethylene glycol in the confine of a fume hood. Additionally, we designing a circuit in which produce enzymes that will degrade ethylene glycol into glycol aldehyde and then glycol ate. The glycol ate has the potential to be turned in to oxaloacetate, a metabolic intermediate in our device.
      The device that we use in this project are Escherichia coli DH5 Alpha and BL21(DE3). According to Material Safety Data Sheet (MSDS), the wild-type strain of E. coli may irritate the skin, eyes, respiratory tract, blood circulation, and kidney. To minimize the risk, same as the protect needs from ethylene glycol our team and the researchers in our lab always working safely by using gloves, lab coats, mask, and goggles (if necessary).
   2. Risks to the safety and health of the general public (if any biological materials escaped from your lab):
      Our project would not impact the environment in a harmful way. To guarantee the safety and health of general public, we deal with experimental waste and DNA. We also dispose the experimental bacteria, medium, and disposable goods through destruction stages by autoclaving to avoid potential safety hazard.
   3. Risks to the environment (from waste disposal, or from materials escaping from your lab): Ethylene glycol is a clear, colorless, syrupy liquid with a sweet taste but no odor. It has low volatility and miscible with water and some other solvents, slightly soluble in ether, but practically insoluble in benzene, chlorinated hydrocarbons, petroleum ethers, and oils. Ethylene glycol released to the atmosphere will be degraded by reaction with hydroxyl radicals. Ethylene glycol is readily biodegradable in standard tests using sewage sludge. However, ethylene glycol can potentially be toxic within waterways so to ensure environmental safety the team has taken extra precautions to dispose of ethylene glycol safely. In addition all of the hazard waste from our lab is not released directly to the environment.
   4. Risks to security through malicious mis-use by individuals, groups, or countries:
      The hazard posed not only by lab workers as well as the design in our devices but also determined by abuse committed by malicious people. The bacteria we use have special requirements for life. Therefore, these bacteria can not be used freely, especially if somebody want to activate the system that we have designed.
   5. What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.) As detailed above to reduce this risks we have to wears laboratory protector such as gloves, mask, goggles and lab coats. For device that we used is Escherichia coli that very common bacteria for use in the biological experiment, so we have been trained to handle it.


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 themethods you develop became widely available? Our project when properly functioning, should not have any harmful effect on the environment, researcher, or the public. We keep everything contained within lab and all the plates and wastes are autoclaved to prevent all bacteria we use from spreading to the outside environment. Our BioBricks parts this year do not raises any serious safety issues. If our project moved from a small-scale lab study become widely used as a commercial/ industrial product, the risk are kept at a very low level because our Engineered Modified Bacteria only could survive when the environmental conditions are appropriate.
   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. Synthetic biologist, and iGEM teams in particular, design genetically engineered Organisms (GEOs) to benefit people and communities. Our team goal was to break down PET at the source of the problem itself. Our BioBricks parts this year do not raises any serious safety issues. However we are using LC-Cutinize gene(BBa_K936000), encodes an enzyme that breaks down PET into ethylene glycol, a moderately toxic substance. We are also designing converting module which contain Glycol aldehyde Reductase and Glycol aldehyde Dehydrogenase that could convert ethylene glycol become glycol ate so E.coli that we are using as chassis could use it as a substrate for TCA cycle. The engineered bacteria are not designed to be exposed to the general public. We intend to utilize bioreactors to concentrate these bacteria in order to achieve high efficiency. To eliminate safety risk to people around bioreactors, in the future, we are planning to use kill switches that kill escaped bacteria. Horizontal Gene Transfer become the most common risk that could appear from GEOs. Other strain/species may uptake engineered genes, and if the genes give advantage in fitness, it may outcompete other strains creating natural imbalance. So that designing a safety containment to degrade DNA and/or separate the DNA from the environment and/or prevent natural strains from gaining advantages from modified DNA is very important for further development.


6. Further Comments
   If you are completing a Preliminary Version of your Safety Form, use this space to describe how far you have progressed in your project, and give some comments about any questions that you left blank. You can also use this space for any other comments or additional material.
   Currently our projects are already at the stage of assembling parts and test for the expression of several parts. So far our lab safety condition is quite good and can be handled well too.