Team:UNICAMP-EMSE Brazil/Safety

Safety Proposals

1. Would any of your project ideas raise safety issues in terms of:

• researcher safety,
• public safety, or
• environmental safety?

Any kind of research evolves risks, but in our project we tried to assure biosafety with the following actions:

1. All the students were trained by the advisors (PhD students with experience in molecular biology and microbiology).
2. Every lab practice should be performed with gloves, labcoat and other safety equipment if required.
3. We are using non-pathogenic lab strains of E. coli that do not confer any additional fitness advantages on bacteria, apart from the most common antibiotic resistance genes used as selection markers. We are focused on containment of this microorganism by discarding all the genetic or microbiological material after sterilization.
4. All the genes from pathogenic bacteria were synthesized to avoid safety problems with environment, public and researches health.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,

• did you document these issues in the Registry?
• how did you manage to handle the safety issue?
• How could other teams learn from your experience?

• There's no directly evidence that the biobricks developed by our team should raise any safety issue, besides in the manipulation of GMO and lab practices. But if we encounter any reason for this kind of concern, we will report and document it.

3. Is there a local biosafety group, committee, or review board at your institution?

• If yes, what does your local biosafety group think about your project?
• If no, which specific biosafety rules or guidelines do you have to consider in your country?

• The wetlab work of team UNICAMP-EMSE was conducted at the [http://sysbiol.cbmeg.unicamp.br Systems Biology Laboratory], located at Center for Molecular Biology and Genetic Engineering (CBMEG) in [http://www.unicamp.br/unicamp/en University of Campinas - UNICAMP]. Systems Biology Laboratory is a research laboratory fully equipped for dealing with Risk 1 microorganisms and GMOs. The following laboratory equipments are available: autoclave for sterilization; laminar flux hood with UV lamp for working with microorganisms under sterile conditions; shakers and incubators for microorganism growth; freezers and refrigerators exclusively used for maintaining culture media and bacterial cultures; specifically designated sharps collection bin; easily accessible instructions on how to deal with contamination events; and log book for recording spills and contamination events. The laboratory has doors with a biohazard sign and access to a specifically designated culture room is restricted to trained personnel. The culture room is provided with a sink and has a controlled environment, without opened windows; its tiled floor can be easily cleaned, and the bench tops are impervious to water and resistant to moderate heat and moderately corrosive organic solvents. Insect and rodent control programs are in effect at UNICAMP.

• In Brazil we follow the guidelines instituted by CTNBio (National Technical Commission for Biosafety) and CBMEG is certified to work with GMOs (CQB 0086/98, http://www.ctnbio.gov.br/index.php/content/view/2267.html?id=09&op=2).

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

• One suggestion in the biosafety of the researcher is to include in the iGEM parts kit the bacterial less endotoxicchassi (developed by Berkeley UC 2007), to avoid serious septic problems in the case of an accident that leads to an intense contact with the bacteria (eye or bloodstream contact, inhalation or ingestion).

• Another suggestion for population and environment safety is to (somehow) include in all biobricks an operating unit that detects if the bacteria is not in a culture media (detects some molecule produced through the metabolism of a specific media constituent) and express lysozyme to kill it if it's released in the environment (idea inspired by Team UNICAMP-Brazil 2009).

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? b. Risks to the safety and health of the general public if released by design or accident? c. Risks to environmental quality if released by design or accident? d. Risks to security through malicious misuse by individuals, groups or states?

• As far as we know, there are no risks related to our biobrick. Our model is a bacteria, and we will take every care so that it doesn’t get released by accident, but it is not more harmful then any other laboratory E. coli. It is not pathogenic and doesn’t produce toxins or any substance known to risk the health or the environment. Furthermore, we do not expect our biobrick to improve or facilitate the bacteria’s survival.
• But if for any reason our bacteria is released into environment, we could also build a self-destruction mechanism of the bacteria, this is interesting because could avoid the genetic exchange with other bacteria found in the environment. This mechanism could be built in a very similar to that developed by the UNICAMP team from 2009 called “The kamikaze system”. This mechanism consist in the production of a huge amount of lysozyme by the killing cell, this lysozyme in high concentrations will be able to attack the cell wall of E. coli passing trough the inner cell membrane. This will destroy the E. coli releasing lisozyme in the medium.
• In future our bacteria maybe be used like a probiotic, however studies are in "embryonic" phase and in very early stages of development. So, the bacteria carrying this device should not be consumed.

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.

2. If your response to any of the questions above is yes:

• Not applied.

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.

• No, our institution follows the national rules.

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, our institution has a internal committee dedicated to biosafety, which is responsible for evaluating and deciding the laboratories that are allowed to work with GMOs, including ours. No suggestions or requirements were done since our project fits in the specifications required by them.

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

• Yes, our advisors and students have prior experience in molecular biology and have all been trained to pay attention to biosafety during their previous lab experiences.

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

• http://www.ctnbio.gov.br/index.php/content/view/12847.html
  [http://www.ctnbio.gov.br/index.php/content/view/12847.html CTNBIO] is a committee composed of government members designed to discuss the law applied to biosafety issues as well as to inspect the application of these laws.

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?