"
Team:INSA Toulouse/contenu/safety/safety in the lab
From 2013.igem.org
Safety in the Lab
Safety in INSA Toulouse
INSA Toulouse is a public engineer school in which several government research laboratories are present. Workers at INSA must follow the French rules concerning safety and environmental regulations. Most of these rules are presented in the manual “Manuel de sécurité en laboratoires” (from the WHO), which allowed us to understand and apply safety rules when manipulating microorganisms and chemicals. In addition, daily use of the Deming wheel allowed us to improve the safety rules and keep them updated.
Concerning the laboratory
Personal protective equipment
We used conventional protection all along our daily manipulating. All experiments were perfomed with gloves and a conventional lab coat, protecting the worker and avoiding contaminations. We also wear glasses when needed (UV exposition, hot water manipulations, chemicals manipulation, etc.).
Biological and Chemical Waste
After a 1h30 training for chemical waste gesture and biological hazard, we learned how to use different trash containers as described in the picture. Biological waste, potentially containing viable microorganism, were autoclaved in a special bag, then discarded. Chemical wastes are grouped by categories and eliminated via the regular procedure for all chemicals of the INSA.
Devices and Material
Here is a list of devices we used and how to use them safely:
- Chemical Storage We got separated and dedicated racks for every kind of chemical product we use. As you can see on these pictures, those boxes are key-closed.
- Ethidium bromide A dark room is dedicated for using EtBr and UV. This room is key-closed. Everyone entering the room must wear gloves, glasses and lab coat. Two special boxes are available to discard spoiled EtBr (one for used agarose gels, a second for gloves).
- Biological safety cabinet To avoid external contamination by unwanted microorganisms on agar plates we used a Biological safety cabinet (FASTER – Ultrasafe). The cabinet is cleaned-up every morning when we arrive and every evening when we leave.
- Electric burner For all sterile manipulations, we used electric burners. They are as efficient as Bunsen burners (even more), but much safer than them (limiting greatly the risks of fire hazards).
- Chemical Hood To protect the user from volatile chemical compounds while dangerous manipulations, we used of two chemical hoods.
- Water-bathes Use of water-bathes can be dangerous, with exposition of boiling or hot water. We used special gloves for protecting us from projections and steam. The use of water-bathes implies of course wearing glasses and lab coat. Special care was taken to switch them off every evening before leaving!
- P2 Laboratory Special care have been taken to manipulate the Chromobacterium violaceum strain which is classified as a class 2 microorganism. All manipulations with this bacteria were performed in a a dedicated P2 laboratory, fully equipped for microbiology. Biological waste from C. violaceum were treated with special procedures according to the Safety Regulation Rules for Class II microorganisms.
Concerning our E. calculus project
Would E. calculus raise safety issues in terms of:
- Researcher safety? In this project, we are working with biosafety level 1 microorganism (according to World Health Organization (WHO) (Laboratory Biosafety Manual) with Escherichia coli DH5-alpha, DH5-1 and XL1-Blue strains, which are considered as biologically safe for users. Some of chemical compounds are dangerous for health (EtBr, Phenolchloroform, Chloramphenicol), but as previously described, all precautions are taken to ensure security and safety for all users.
- Public safety?
As described in the overview, E. calculus is not designed to be used on other environments than the laboratory. Interactions with human body are not envisaged and public should also never be in contact with our modified organism. All the experiments we have performed we made according to the highest standards of safety measures. Each modification we apply to our strains should not be threatening for biodiversity and external organisms beacuse the E. coli strains that are routinely used in the laboratories are mutated in many genes, and should not be able to survive in an external environment.
However, we are conscious that the gates we designed could potentially (and hopfully :-)) used for other biological applications, especially in metabolic engineering. In our view, these potential applications will probably be part of a process in a closed reactors, or in controlled environment, hence the safety regulations in use in the industrial context will apply. We also believe that our system present no evident potential danger, mainly because the used genes or parts were never described as potential threats to human and hopefully will never be an issue for public safety. - Environmental safety? E. calculus should not be disseminated in the environment of course. Avoiding contaminations and accidental release of the E. coli strain implies good laboratory practise, particularly for elimination of biological waste (see above).
Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?
The only safety issue we can associate with our new Biobricks is researcher exposure to a microorganism when testing the parts. During the iGEM summer work, all safety measures were respected to avoid any problem.
Is there a local biosafety group, committee, or review board at your institution?
There is one biosafety group, leaded by a specialist in safety regulations in our institute. This person is in charge of the evaluation and implementation of the safety measures in the Instititute. Moreover, as INSA Toulouse is a five years engineer school, there are plenty of review boards that we can consult at any time to answer arising questions about biosafety. In addition, biosafety issues are regularly evaluated by the security laboratory officer.
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?
Safety issues are usually highly specific for each project. Associating DNA parts together will always pose safety risks in case of accidental release of DNA or bacteria occur. European safety rules are pretty strict and are tailored to avoid accidental contamination of the environment or the humans manipulating these microorganisms. We think that the fact that Safety Issues are mandatory for iGEM projects is a very good way to force students (and their advisors!) to take some time evaluating (or getting help to evaluate) their specific safety (biosafety) issues.
When we discussed biosafety issues of our project, we analyzed the 2012 Paris-Bettencourt iGEM project named "Bware", which proposes several measures to ensure that new synthetic constructions do not consitute health or environmental issues if the strain was released on the wild. Such a system would be ideal to destroy any genetic material that could be released during or after the use of E. calculus.
