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From 2013.igem.org

Contents 1 Biosafety Course 2 Safety 2.1 Form 2.2 Hazard

Biosafety Course

Before performing experiments, we were invited to know more about the lab routine and the procedures required to cope with organisms genetically modified and compounds usually required to perform experiments as well. Everyone interested in performing experiments during the competition was invited to a course to receive instructions and to learn more about the biosafety and its implications on lab working.

The central idea of this course was discuss about what is biosafety, why this is important and what is its implications on lab working. Under this perspective we started reading several texts to increase our knowledge about the subjects listed below :

The book Manual de biossegurança[1] (Biosafety Manual), a collection of texts written focused to teach the reader the most important procedures for control, handling and discard of biological products, for avoiding experiment contamination and the vigent law (at least the main) related to manipulation of biological organisms in teaching and researching as well. It is a very detailed book, that helped us to get a notion about where we are inside this big area named biosafety.

Embo Reports including Science and ethics[2] related to ethics, since we proposed a mechanism to detect biomarkers in a human serum in order to diagnose heart diseases. It is known that we cannot simply perform any sort of experiments using wild animals or even human to look for results that corroborates hypothesis of a work. But in many cases people do not know what is the correct behavior to use samples gotten from human being or from wildlife for research purposes. That is, until where we can go with research in a way that do not harm the species analysed in the experiment, mainly humans? Each country have its culture and people may have different interpretations of what actions can cause ethical problems[2]. For this reason and many others, ethics is so debated in research. There must be a common point for everyone in ethics on researching?

Crowdfunding society involvement and genetical engineering consequences. Recently, Callaway group created a plant with ability to glow in dark[3] and they were sponsored by people using a mechanism named crowdfunding. The intense interest of common people about this plant cause a general commotion in scientific mean about the consequences of spreading such modified organism unsupervised. It is not easy to predict the behaviour of such organisms into nature as well as its interactions between other organisms in environment. Since the genetical engineering grew as a big research field, many things emerged providing improvements to health (as the production of insulin), to nourishment (production of soy resistant to plagues) and energy (production of biofuels), for example, but all under restrict safety control.

During the course, we talk about such events and we comprehended biosafety not just as a list of rules that must be followed. Beyond of all restrictions applied to ensure safety, biosafety should be understood as what do you do not want to carry to your friends, relatives or all sort of people you know in order to keep them safe of any kind of risks. It is more related to avoid risks of being carried from lab to the environment than just hold them in a safe place. In the end we also concluded that we must have our critical sense always keen when dealing with science, mostly with life.

References:

• [1] Manual de biossegurança/Biosafety manual, Hirata, Mario H., Hirata, Rosário D.C., Mancini Filho, Jorge, 2012
• [2] Science and ethics, Iaccarino. M, Nature - EMBO reports vol. 14, September 2013, doi:10.1093/embo-reports/kve191
• [3] Glowing plant spark debate, Callaway, E, Nature 498, 2013 June 06, doi:10.1038/498015a

Safety

Form

Safety forms were approved on September 24, 2013 by Evan Appleton.

Basic Safety Questions for iGEM 2013

1a. Please describe the chassis organism(s) you will be using for this project.

Species: E. coli K-12 Strain no/name: XL1-Blue Risk Group: 1 Risk group source link: www.absa.org/riskgroups/bacteriasearch.php?genus=&species=coli Disease risk to humans? If so, which disease? Yes. May cause irritation to skin, eyes,and respiratory tract, may affect kidneys.

2. Highest Risk Group Listed: 1

3. List and describe all new or modified coding regions you will be using in your project. (If you use parts from the 2013 iGEM Distribution without modifying them, you do not need to list those parts.) We did not use new modified coding regions, we only used new promoter regions.

4. Do the biological materials used in your lab work pose any of the following risks? Please describe.

a. Risks to the safety and health of team members or others working in the lab? Our constructs are based on E. coli and they do not offer any hazard beyond the ones intrinsic to the microorganism itself.

b. Risks to the safety and health of the general public, if released by design or by accident? The E. coli strain used in this work is not harmful to human health and has not any new genetic material that makes it harmful or gives evolutionary advantages.

c. Risks to the environment, if released by design or by accident? No new genetic material was added to the bacteria that gives it evolutionary advantages or makes it harmful if released into the environment.

d. Risks to security through malicious misuse by individuals, groups, or countries? Our engineered bacteria will be able to detect and quantify biomarkers for prognosis of cardiovascular diseases. However, several tests must be completed before we can validate the accuracy of this use. Thus, until that, our bacteria must not be used as a primary way of detecting cardiovascular disease.

5. If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? (Consider the different categories of risks that are listed in parts a-d of the previous question.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? (Note: This is meant to be a somewhat open-ended discussion question.) Since our E. coli does not carry any new coding region, even if it becomes an industrial product it will not offer any hazard beyond the ones intrinsic to the bacteria.

6. Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis, etc.). Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them. As of now, our project does not include any design feature for safety risk, but we plan on including one in the future.

7. What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution’s safety training requirements, if available. All participants of our team had to have taken a course on biosafety to work in the lab. Some had already done the course and others have done especially for this project. The course included basic rules of laboratory procedures on the containment of GMOs and the study of legislation on the subject.

8. Under what biosafety provisions will / do you work?

a. Please provide a link to your institution biosafety guidelines. http://www.icb.ufmg.br/cibio/site/wp-content/uploads/2013/03/resoluo-normativa-no-2-de-27-de-novembro-de-2006.pdf

b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project plan based on their review. Our institution does have a Biosafety Committee. We have discussed our project with the person responsible for the biosafety course in the university, Neuza Antunes. She just highlighted the importance of following the Committee’s biosafety guidelines. c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible. Yes. We have an institution called CTNBio that is responsible for normalization of procedures that deals with genetically modified organisms. http://www.ctnbio.gov.br/upd_blob/0001/1620.doc.

d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? (Check the summary table on page 3, and the fuller description that starts on page 9.) If your lab does not fit neatly into category 1, 2, 3, or 4, please describe its safety features. Our lab is level 1 for most of its space, and one specific room is level 2.

e. What is the Risk Group of your chassis organism(s), as you stated in question 1? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking. Risk Group 1. No additional safety measures were necessary in this project.

Hazard

As described in “Safety”, the risks involved with the bacteria that we used in our experiments are only the ones intrinsic to this microrganism and they are listed in the section former mentioned. To avoid these risks, we have followed biosafety guidelines. Cobalt is a toxic compound, if you are constantly exposed to large amounts of it. It can cause cardiomyopathies and nerve and thyroid problems (http://hazmap.nlm.nih.gov/hazardous-agents). As we used personal protective equipment (PPE) on its manipulation, and small quantities were used, risks involved with cobalt were understated. TMAO may cause skin, eye, and respiratory tract irritation. It is safe when used as a flavoring agent in food, but it is a strong skin and eye irritant (http://hazmap.nlm.nih.gov/hazardous-agents). As for cobalt, we used PPE and only small amounts of TMAO, what reduced its manipulation risks.

Reference:

HazMap. http://hazmap.nlm.nih.gov/hazardous-agents.

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