Team:Groningen/10com

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Revision as of 11:27, 6 September 2012

Working with genetically modified organismes (GMOs) and microbiology is obligated to be done under safe conditions. The European union made a directive that insures the safety for the researcher, the laboratory and the environment. The directive however is very comprehensive and very detailed. To make a good and safe work environment the University of Groningen (RUG) made document listing all rules, regulation and organisational structueres regarding to the work with GMOs at the university. This document is called the ’Administratieve organisatie GGOs’ and is based on the EU directive ‘98/81/EG’ and was approved by the ministry of Infrastructure & Environment from the Dutch government.

The government is in turn advised by the Netherlands Commission on Genetic Modification (COGEM), the scientific advise organ of the government.
The COGEM provides scientific advice to the government on risks to human health and the environment of the production and use of GMOs. The organisation consists members appointed by the Minister of Infrastructure and Environment. All members are selected for their scientific expertise and do not represent any organisation. The members have expertise in different fields such as ecology, bacteriology, virology, genetically modified plants and animals, zoology, public perception and ethics. More information on the COGEM is found on the COGEM website

Students studying microbiology at the University of Groningen are informed on safety aspects of microbiology from the beginning of their study career. They are required to get a VMT (Veilige microbriele technieken, translated: safe microbiological techniques) certificate that demonstrates that they are able to work safely with microbiology and GMOs. The bases for this certificate are the 10 commandments for safe microbiological techniques which are derived from the Administratieve organisatie GGOs. Every iGEM member of the iGEM Groningen 2012 team is required to know these rules and uphold them when working in the lab. These commandments are 10 rules which ensure safe work environment for researchers on a daily basis. Most of the team members that work in the lab have a background in microbiology, taught at the RUG. These persons have the VMT certificate and know how to work safely in the lab. Team members without a background in microbiology, and who work in the lab, have been instructed with these rules and learned how to work safely.

10 COMMANDMENTS for Safe Microbiological Techniques

1. All VMT related work can only be performed by those people that have permission from the Biological Safety Officer (BVF). Work according to the rules, even if you believe there is no apparent risk.
2. During VMT related work all doors and windows have to be closed. Verify that insects and other pests are not present in the lab.
3. Wear a closed laboratory coat. Do not take this labcoat outside the VMT area unless it is absolutely necessary for the experiment. In case of a contamination of the labcoat, sterilize the labcoat first, with bleach or by autoclaving, before washing.
4. Clean and sterilize spills immediately. Report serious contamination immediately to the BVF.
5. It is absolutely prohibited to eat, drink or smoke, or to have cups, plates, mugs or silverware in the VMT area.
6. Pipetting by mouth is prohibited. Used pipettes are collected in a disinfecting solution.
7. Prevent aerosols. These may be created by splashing drops, pouring of liquids, discharging pipettes, opening of moist plugs, using inoculating loops that are too hot. Use needles only if there is absolutely no alternative.
8. Glassware and instruments that have been in contact with GMO's (Genetically Modified Organisms, GGO’s in Dutch) have to be sterilized or disinfected before being washed, reused or discarded. Biological waste has to be collected in autoclavable plastic bags, which are autoclaved before discarding (use indicator tape to demonstrate that the bag was autoclaved).
9. Wash hands with soap and water after work and before leaving the room. Bench surface areas have to be cleaned and disinfected daily. Keep area clean and organized.
10. Record the general nature of the work clearly in a lab journal.

More to consider


Ethidium bromide is a carcinogenic compound that we used when working with agarose gels. To ensure our safety, we created a specific ethidium bromide region in our lab; in this area is clearly indicated that it is obligatory to use appropriate gloves and keep all the contaminated equipment in the EthBr area. The toxic waste is disposed in the appropriated manner according the rule and regulations uphold in the laboratories of the university.

Micro-organisms involved


We worked with rotting meat, a versatile environment for the growth of Pseudomonas, Salmonella, E. coli and other harmful bacteria. In order to work as safely as possible, we performed our rotten meat experiments in closed bottles and used our flow cabinets and procedure masks when taking meat samples. All of the meat was assumed to be biological waste, so we put everything in the autoclave to ensure that our researchers worked as safe as possible. You can read more about food safety in the dedicated chapter below.

Bacillus subtilis. For proof of principle we used B. subtilis strain 168, as safe strain widely used in labs. For our final product we want to use a food grade B. subtilis strain as Bacillus subtilis natto. Considered as safe as Lactococcus lactis in yoghurt.

Escherichia coli strain DH5α- Standard laboratory bacterium, widely used as chassis for synthetic biology. This bacterium is considered moderately safe, however, may cause infections and therefore has to be treated carefully.

BioBricks


We did not find urgent safety issues with our biobricks. The promoters were taken from the genome of B.subtilis 168 and are natural gene parts. Furthermore the other parts of our biobricks are parts taken from the biobricks in the registry. The Lycopene pigment is considerd food grade and thus safe. GFP and the chromoproteins used as reporters some of our constructs are widely used reporters that are considerd low risk, however we do not plan to use that in our final Food Warden system and are only used under safe conditions in the lab. Furthermore we are planning to make our final Food Warden construct without an antibiotic resistance marker.

All our biobricks contain simple modification that have low or no risk at all. So under normal working conditions in the lab, these biobricks should not pose any danger to the researcher.

Biosafety group


Every research facility has staff responsible for the safety of the labs where people work with GMO. The University of Groningen is no exception. There are four biological safety officers each charged with a different aspects of safety (Site manager, Microorganisms, ecology and plants, animal safety) We invited Dr. Juke Lolkema, the safety officer in charge the safety in working with microorganisms, to give a lecture about GMOs and safety in the lab. Dr. Lolkema is responsible for the maintenance of safe microbial techniques in the whole life science building (Linneausborg)of the RUG. In this lecture we given information about rules and regulation of working with GMOs and how to work safely. During this lecture we asked about the safety aspects of our project. He found no apprend safety issues, due to the nature of our GM insert and the strain of B. subtilis we planned to use. However he raised an issue we needed to address: how to kill the bacteria after use? We tried to come up with ideas to adress this problem and found a good solution: a internal kill switch that is triggered by the a strong promotor of for example a stress factor or sporulation gene. More detail on this solution are found on the evironmental safety page.
After the lecture Dr. Lolkema made an inspection of our lab. No possible safety problems were found.

The safety lectures by the experts provided us with a good understanding into the safety of working in a laboratory. By working in accordens to the safety rule and the use of good commen sense, we provided a safe working environment for the researchers on the 2012 Groningen iGEM team.