Team:DTU-Denmark/Methods/Safety

Biosafety vs. bioethics - who cares and what is the difference?

WE CARE… At the 2014 DTU iGEM team we believe that it is our responsibility as researchers to ensure that the Genetically Modified Organisms (GMOs) we create and handle on a daily basis do not cause any harm to the health of people nor harm the environment. Synthetic biology offers a lot of great opportunities, but also includes handling of different hazards that the scientists need to be aware of to be able to minimize potential risks. We have therefore considered both the biosafety as well as the bioethics of our project. We have chosen to distinguish between the two according the the definition given by the World Health Organisation:

• Biosafety: “The prevention of unintentional exposure to pathogens and toxins, or their accidental release, whereas biosecurity is the prevention of loss, theft, misuse, diversion or intentional release of pathogens and toxins.”
• Bioethics: ‘’The study of the ethical and moral implications of biological discoveries, biomedical advances and their applications, as in the fields of genetic engineering and drug research.’’

Below, we have explained how we have implemented safety in our daily wetlab work to avoid any release of GMOs, hazardous chemicals etc., as well as to ensure the safety of the researchers (ourselves) when handling the biological as well as chemical agents. If you want to know more about our considerations on bioethics, please visit the Ethics section.

How we handled the safety

When carrying out an experimental project, safety precautions regarding the work routines in the laboratory need to be considered to avoid personal injuries or leakage of (potentially) dangerous biological or chemical agents to the environment.

All members of the team that have been actively involved in the laboratory work have received a general safety instruction to the laboratory, in which we have carried out the main part of our research for the iGEM project, by team advisor Ali Altintas prior to starting the work. We also identified when to use personal safety gear such as gloves, safety glasses etc.

In Denmark all laboratories are classified according to the organisms that are handled and different requirements apply to the different laboratories. At DTU, local safety groups are responsible for the safety in the laboratories in the different research groups and departments. These groups specify the rules that every researcher need to follow during their work, and the laboratories are frequently checked by the Danish Working Environment Authority.

On top of the aforementioned division of laboratories into different classes, the laboratories we worked in was also divided into different areas according to what to work with: LAF-bench for sterile work, fume hood for handling of volatile and organic solvents, an area for gel-electrophoresis and gel-analysis due to the involvement of ethidium-bromide etc.

Knowing your organism

GMO or not, you should always know the characteristics of the organism you work with. Is it a human pathogen, does it produce any harmful secondary metabolites, is it a GMO etc.? These are all relevant questions that need to be addressed prior to entering the laboratory.
In our project we work exclusively with E. coli. The strains we work with are GMOs but with a generally regarded as safety status, meaning that the health risks involved in working with them are minimal. Furthermore these strains will not survive well in nature, but extensive care should still be taken to ensure that they do not escape the laboratory.

Thus the main safety concern when working with these organisms is to keep all GMO material inside GMO designated areas (i.e. the laboratory). Rule number one in preventing GMO escape to the environment is to carefully wash hands before leaving the laboratory and never wear lab coats outside the lab area. Furthermore overcoats, cell phones, bags etc. are not allowed in any laboratory.
Another concern is the disposal of GMO waste. All trash that has been in contact with GMOs must be thrown out in special GMO trash containers. The bags from these containers are emptied regularly into locked containers from where they are transported by special transports to get properly disposed of.

Liquid cultures of GMO are disposed of in separate containers and treated with perfektan. The concentration of perfektan needed and the exposure time of perfektan to the liquid GMO cultures depends on the culture mix. Specific instructions can be found here. After successful treatment with perfektan the liquid can be disposed of in the sink.

Additionally, the areas in which the work with GMO is carried out should be cleaned meticulously with 70% ethanol. This is both to avoid the GMO from spreading but also to avoid cross-contamination experiments carried out in the working area.

Knowing your chemicals

Most of the chemicals involved in our experiments are harmless or only dangerous if they come in contact with eyes, are swallowed etc. However, one frequently used chemical that deserves extra attention - ethidium bromide. This compound is used for gel electrophoresis, and is carcinogenic. Thus, we worked with it in designated areas: at separate benches for gels, where gloves should be worn. Gloves are not to be worn anywhere else in the lab, except when working with other dangerous chemicals, in which case they must be taken off immediately after, and the affected area cleaned with 70% ethanol.

Besides ethidium bromide, known to require extra precautions, whenever starting a new experiment, the material safety data sheets (MSDSs) for the specific chemicals used in the experiments should be looked up, to know how to handle them. When working at Technological University of Denmark, MSDS’s are administered by www.kemibrug.dk, but be aware that your institution probably uses another provider.

After running a gel it is often necessary to cut out specific bands for purification. This requires illumination with UV light. To protect the skin from the UV irradiation, we wear protective masks and gloves. The gloves also protect against the ethidium bromide in the gel, discussed above, and should be thrown out after use. The scalpel is not to be used for any other purpose and should always be handled wearing gloves.

When doing an experimental project, safety issues concerning the work is very important in order to keep a safe working environment for yourself and your colleagues. In addition to document our general as well as project specific precautions we have chosen to use the connection to make develop a laboratory-safety guide in words and pictures. This safety-guide is meant for high-school students, university freshmen and others who might need it, and provides information on how to work safely with GMOs, hazardous chemicals, etc. The development of the safety guide is in nice continuation of our outreach-strategy with respect to the target group – however the guide serves an educational as well as an informative perspective.

Safety in our lab
	In our project we work exclusively with E. coli. The strains we work with are generally regarded as safe (have GRAS status), meaning that the health risks involved in working with them are minimal. Furthermore these strains will not survive well in nature, but extensive care must still be taken to ensure that they do not escape the laboratory. Thus the main safety concern when working with these organisms is to keep all GMO material inside GMO designated areas (i.e. the lab). Rule number one in preventing GMO escape is to wash hands carefully when leaving the lab, and never removing the lab coats from the laboratory. Another concern is the disposal of GMO waste. All trash that has been in contact with GM organisms must be thrown out in dedicated trash pins and from there further on to special GMO trash containers. The bags from these containers are emptied regularly into locked containers outside.
	Besides the organisms we work with a range of chemicals. Most of these are harmless or only dangerous if they come in contact with eyes, etc. One often used chemical that deserves particular mention is ethidium bromide. This chemical is used when doing gel electrophoresis, and is a known carcinogen. We therefore have a separate bench designated for gels, where gloves must be worn. Gloves are not to be worn anywhere else in the lab, except when working with other dangerous chemical, in which case they must be taken off immediately after, and the affected area cleaned. Liquid cultures of GMO are disposed of in separate containers and treated with perfektan (http://www.medicalsupply.gr/dat/43D94425/file.pdf). After running a gel it is often necessary to cut out specific bands for purification. This requires illumination with UV light. To protect the skin from the UV irradiation, we wear protective masks and gloves.