Team:KU Leuven/Safety
From 2013.igem.org
Secret garden
Congratulations! You've found our secret garden! Follow the instructions below and win a great prize at the World jamboree!
- A video shows that two of our team members are having great fun at our favourite company. Do you know the name of the second member that appears in the video?
- For one of our models we had to do very extensive computations. To prevent our own computers from overheating and to keep the temperature in our iGEM room at a normal level, we used a supercomputer. Which centre maintains this supercomputer? (Dutch abbreviation)
- We organised a symposium with a debate, some seminars and 2 iGEM project presentations. An iGEM team came all the way from the Netherlands to present their project. What is the name of their city?
Now put all of these in this URL:https://2013.igem.org/Team:KU_Leuven/(firstname)(abbreviation)(city), (loose the brackets and put everything in lowercase) and follow the very last instruction to get your special jamboree prize!
![tree](https://static.igem.org/mediawiki/2013/a/ac/Tree.png)
![ladybugcartoon](https://static.igem.org/mediawiki/2013/6/64/Ladybugcartoon.png)
How have we tried to make our project as safe as possible?
We have covered the safety aspect of our project in many ways. Within synthetic biology there are three main risks that have to be covered: exposition of the laboratory workers to the bacteria and chemicals, effects of a possible escape of the bacteria and the unintentional disturbance of the ecosystem.
We received extra safety training at the beginning of the project. On top of that we took courses regarding safety aspects, e.g. ‘Safe Microbiological Practices’. The safety training consisted of a summaryof the important biosafety issues and the composition of the lab.
![Laurens Labo](https://static.igem.org/mediawiki/2013/6/66/Laurens_labo.png)
![some_text](https://static.igem.org/mediawiki/2013/f/ff/Ladybug_full.png)
(Max Gibson)
In the past three months, we tried to add new branches to the pathways of E. coli. Our experiments were performed with non-pathogenic E. coli strains (BL21, DH5α, TOP10 and Rosetta(DE3)pLysS), which belong to risk group 1of the pathogenic organisms. Working with genetically modified organisms needs extra safety conditions, physical measures are taken to separatethe organism form the surrounding environment and vice versa.
Our project wants to protect plants and crops against aphid infestations and reduce the use of harmful insecticides. Our genetically modified bacteria will produce (E)-β-farnesene (EBF) and methyl salicylate (MeS), which are natural occurring substances. EBF repels aphids and MeS attracts ladybugs, one of their natural enemies. The low concentrations needed to be effective are not toxic for humans. No harmful effects are described for EBF at high concentrations either. Contrarily, pure MeS could cause harm when it is ingested due to its similarities with acetylsalicylic acid (aspirin) and can cause irritation after contact with eyes and skin. Although, it is typically used in small amounts as a flavouring agent, in candy or as fragrance oils due to its mint flavour and wintergreen fragrance. The two chemicals being produced by our BioBricks could cause allergic reactions (swelling, redness, ...), but serious reactions are very rare.
The effect of our genetically modified bacteria on the environment is non-trivial and thorough field experiments must definitely be conducted. The maximum production rate of onebacterium has to be determined and scaling this model up to a whole field occupied by our bacteria will allow us to estimate the concentration in the surroundings. We can then determine the necessary parameters in order to minimise ecological disruption according to surrounding concentrations of MeS and EBF. We also thought about the interactions between aphids and ladybugs themselves. Aphids adapt readily when EBF is constantly present,therefore we included an oscillatory model. More information about this model can be found here .
General safety
Legislation in Belgium & Europe
![some_text](https://static.igem.org/mediawiki/2013/3/3e/European_figure.jpg)
(JRC - European Commission)
University of Leuven Rules
The University of Leuven has its own biosafety rules in addition to the national biosafety regulations, where Marianne Schoukens acts as the central contact point for Health, Safety and Environment (HSE) issues. All members of our team have received training in Safe Microbiological Techniques as well as general and specific lab safety required by the University of Leuven. At the beginning of our project, all members of the iGEM team have received an update from the KU HSE department. Via liaison Iris Govaerts, we followed a small seminar and filled in an orientation checklist concerning health and safety training prior to the start of project laboratory work.
The project we ultimately chose was the result of a whole series of brainstorm sessions. In this brainstorm sessions we talked about the feasibility of our ideas with our instructor (Dr. Ir. Ingmar Claes) and our coordinators (Prof.Dr. Johan Robben and Dr.Veerle De Wever). We also invited different PhD students, professors, ex-iGEMers and people from the industrial sector to our brainstorm sessions. These people have a lot more knowledge about the specific topics and could help us greatly with our decisions. Once the project was in progress, we also contacted a lot of people with questions and about possibilities. All these people had another view on our project and helped us taking into account different issues about the safety of our project: the molecules that our bacteria will produce, release into the environment, etc ...
Environmental safety
If our project would be used around the country or in the whole world, many parameters would need to be determined. As stated earlier, the pheromones produced by the modified E.coli are not harmful for our health unless available in high concentrations. So when this modified E.coli would be spread across acres of land, the concentration of pheromones could attain a high level. The ceiling of this production would need to be determined to see whether it could be a harmful concentration for our health. Insects however do not need as high of a concentration as humans do to develop a reaction. Therefore the disruption of the ecosystem would need to be considered as well. As with most products released in the environment, its bioaccumulation needs to be at a minimum.
![](https://static.igem.org/mediawiki/2013/e/ec/Ladybug_tree.png)
(Norcal)
Bacterial safety
In all our experiments, we used non-pathogenic E. coli strains. These strains are not harmful for humans, except when they are inhaled or swallowed. Nevertheless, they can cause irritation to the eyes and/or skin. The chemicals that are produced by our genetically modified E. coli strain are natural molecules and are not harmful for the environment. An accidental release of our bacteria into the environment could have unexpected effects on the ecological balance.As suggested in literature (reference), containment through engineered auxotrophy so that our genetically modified E. coli is unable to synthesise an essential compound required for their survival, is the most reliable method currently for biocontainment. We propose the use of a tryptophan auxotrophic bacterium.
When tryptophan auxotrophic bacteria will be used in our project, we must provide tryptophan in their medium. This is possible when the bacteria grow together on a petri dish or another closed environment, but not when we want to spray the bacteria on the plants and crops. Therefore we designed two models. In one model the bacteria are grown in a contained environment, a plastic sticker (Design of Groningen iGEM 2012), where the auxotrophic mechanism can be used. This plastic sticker is composed of a plastic film with pores that are too small for the bacteria, but through which our volatiles (EBF and MeS) can escape.
![some_text](https://static.igem.org/mediawiki/2013/5/5d/Stickermodelverkleind.png)
![some_text](https://static.igem.org/mediawiki/2013/f/f2/Spray_antiaphid.png)