Team:UiOslo Norway/Safety
Biosafety
The UiOslo iGEM team and our project have been governed by the general regulations and safety procedure of the University of Oslo, regulations that apply to the institution as a whole. This is to ensure a good work environment and the safety of the employees and other members of the institution.
As a researcher you have a responsibility that your research is performed according to the standards of good research practice and established scientific and ethical principles. UiO has a strong focus on research ethics and safety. Read more information of UiOs research ethics.
The safety guidelines and teaching in laboratory is specialized according to which department they belong to. UiOslo iGEM belongs to the faculty of Mathematics and Natural Sciences, at the Department of Biosciences. Our lab is located in Kristine Bonnevies house, 3rd floor, roomnr: . Responsible for this lab is our supervisor Dirk Linke. This is a BSL2 lab and its specific regulations are listed.
The specific guidelines and local HSE procedures and tools that UiOslo iGEM complies with are available online and linked.
Biosafety Training
All students at the UiO receives HSE training specialized to the faculty they belong to. These are mandatory courses starting up during the first weeks of every semester. This kind of training ensures that all students, as well as other members of the working environment, have the necessary training to perform for their duties and fulfill their responsibilities put forward by their work.
In addition to this the UiO iGEM team has been through a customized HSE review specialized to the lab where our workspace is located. This review includes risk assessment, appropriate use of personal protective equipment (PPE) and proper waste disposal. As already mentioned, it is a BSL2 lab, meaning that all of these things are of utter importance. Also, of course, over the course of the last months, we have developed some routines to ensure that the safety guidelines are followed at all times.
First of all we never work in the lab alone; there have been a minimum of two or more students from the team at all times. In addition to this, a responsible from our supervisors research group have been available and nearby at all times. This means that in the case of something going awry, there have always been experienced personnel in immediate distance. As some of the team members are not molecular biologists and have limited lab experience they have been encouraged to join lthe ab work with one of our molecular biologists, who are used to functioning in the landscape of the lab. In this way we have ensured that safety measures have been maintained at all times, and we have never felt uncomfortable in the lab.
Health risks and personal safety
Working in the lab, especially in a BSL 2 lab, came with specialized guidelines and regulations. To minimize the risk that may arise from working in such a lab, it is important to always be aware of the risk factors every experiment pose, in addition to those that may arise if something goes wrong. We have assessed these risks and taken measures to minimize them. When working with bacteria it is important to wear gloves and an appropriate labcoat at all times to minimize the chance of contamination. Before any experiment, the appropriate SOP must be carefully read through. UiO has a library of common procedures that is performed at the Department of Biosciences; here we obtain the appropriate SOP.
In the cases where an appropriate SOP did not exist in this library, a safety representative from the team wrote an appropriate SOP and had it approved by the HSE manager. A SOP (procedures, risk assessment and waste disposal) informs of the risks that comes with the experiment in question and how to minimize these, what the researcher should be aware of when performing the experiment and how to appropriately dispose of reagents and waste.
A list of some of the precautions the team have taken for regular procedures in the lab is listed here:
* When working with bacteria gloves and lab coat are used at all times.
* Agar plates with bacteria are completely sealed when set for storage, thus preventing their escape.
* Transfer of bacterial cultures to or from agar plates/petri dishes is performed in fume hood with appropriate ventilation minimizing the possibility of spread and contamination.
* Specialized risk waste is used when disposal of bacteria and equipment contaminated with bacteria.
* Always have 70% ethanol available for quick disinfection if spillage occurs.
Working with organisms in risk group 2
Working with organisms in risk group 2 demands higher safety and more attention to possible risks compared to organisms in risk group 1. Biological factors in this context is living or dead microorganisms, cell cultures, endoparasites and prions that has the possibility to evoke infections in humans. Read morea about the specified procedures and regulations for working with biological factors class 2 from our university.
These regulations are in Norwegian, and contain procedures on how to work with biological factors in a secure and healthy manner. Biological factors class 2 should be treated with caution because of their infectious potential. For this reason, all work that includes biological factors class 2 should occur only in a biosafety 2 level lab with an updated risk assessment, so that the researcher at all times can take steps to reduce the exposure to a minimum. Specific cleaning measures should also be taken with class 2 biological factors. Workbenches should be cleaned and disinfected before and after use. The benches should also be regularly disinfected with a suitable chemical or UV – light. This is to prevent transfer of potential infectious material the spread of this into the environment.
Our work with organisms within risk group 2 was reduced to a minimum. Before starting up with the laboratory work our team had a dialogue with the safety committee from iGEM HQ about the use of antibiotic resistance genes and bacteria harbouring said genes. We recognized the potential risks of working with antibiotic resistance genes, and wanted to get feedback from the safety committees on this. We have also used the check-ins forms provided by iGEM HQ as a precaution even though antibiotic resistance genes do not require a check in. The isolates were obtained from National Expertise Service for Detection of Antibiotic Resistance(K-res), located in Tromsø, Norway. This center has a high level of expertise when it comes to antibiotic resistance bacteria, and their knowledge has been of a huge benefit to us. The isolates has been collected from all over Norway from different institutions. Some of these may be resistant to several types of antibiotics. The isolates was stored in the BLS 2 lab.
Together with our supervisors, we decided to synthesize (by IDT) some of the genes obtained from Tromsø and have them transformed into regular laboratory E. coli. Doing so minimize the amount of work to be done with the actual clinical isolates. Thus, we obtained β-lactamases from three different classes (A, B and C) without working directly with the isolates.
After proving that our test worked and optimizing it using relatively harmless bacteria (for more information about this, see the "Lab" section), we had the test tried out on some of the clinical isolates. This was done under close supervision of experienced lab personnel in our BLS 2 lab. All of the work was of course done in a high security biosafety cabinet using gloves and lab coat at all times. The team had immediate expertise nearby, ready to step in if anything were to happen. When working with these isolates we used disposable tubes that went directly in a risk waste bin designated for work of this nature. When the work in biosafety cabinet was done, UV-light was employed to kill any bacteria inside. In this way, we felt confident that the work was done in a secure manner within a safe environment.
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