Team:BOKU-Vienna/Safety
Safe Lab Work
Safety Course by PI Dipl.-Ing. Dr. Hans Marx
Before being allowed to work in the lab, our PI Dipl.-Ing. Dr. Hans Marx held a mandatory lecture and training on safety precautions in the lab. This included for example clothing requirements, behaviour in case of emergency, emergency contacts, the disposal of chemical and biological hazards, handling of specific lab equipment and the for it required clothing regulations. He also gave us a lab tour to show us all the equipment and how to dispose different chemicals and biological hazards. Every team member who worked in the lab attended this course and had to sign a safety form afterwards. Team members who did not attend this course were strictly forbidden to enter the lab and were given no entrance card for the lab.
Everyday Safety Procedures
Lab coat, lab shoes and lab gloves were always worn. We had a designated lab area for gel electrophoresis to work with DNA staining dye like peqGREEN and SYBR Safe. We did not work with ethidiumbromide, since peqGREEN and SYBR Safe are considered safer. Antibiotics used as selection markers were Kanamycin, Chloramphenicol and Ampicillin. These were ordered and given to us from the Department of Biotechnology (DBT) in powdered form. They were marked as toxic and when diluted, besides wearing the usual lab gear mentioned in the first sentence, safety goggles and face masks were used. We also had designated work areas to dilute antibiotics, which were situated in a different room. Protective gloves were worn when dealing with either too hot or too cold materials (too hot: e.g. media; too cold: cryos and competent cells from the -80°C freezer).
Using Mycolactone and Theophylline
To check if the aptamers for Mycolactone are useful to detect Buruli ulcer, we recieved Mycolactone from Dr. Kingsley Asiedu, medical officer of the World Health Organization. Before that we prepared a report on special safety precautions in regards to working with the toxin. When measuring our genetic system by adding Mycolactone, most importantly a supervisor was always present. In addition to our general lab gear, we wore a second pair of lab gloves over our usual lab gloves and only worked in a designated work area under the laboratory hood.
We used Theophylline as a proof of concept because it was unclear whether we would recieve Mycolactone until very late in our project. Theophylline is a toxin and therefore the abstracted amount (weight) for the experiments was always logged in the "poison book", a notebook to track the use of toxic substances. When working with Theophylline we used, besides the general lab gear, a face mask and safety googles.
Safe project design
Chassis
Our chassis was E.coli DH10B, which belongs to Risk Group 1 and therefore poses no health risk for humans and no environmental risk.
Environmental safety aspect
We thought about the risks posed by our diagnosis method for the environment as well as for the staff using it. Considering them for the staff, the biggest risk would lie in handling samples of patients infected with Mycobacterium Ulcerans.
A risk we could identify, was the antibiotic resistant gene on the plasmid of our chassis. This, by no means, should be contaminating the environment. Thus, we chose to write a manual for the personnel who would work with our diagnosis tool on how to kill the chassis and degrade its DNA. We also talked to experts (see our human practice page) about an auxotrophic marker instead of antibiotic resistance genes. However, due to time constrictions remains as something that should be considered for future experiments in the lab. It would definitely abolish the risk of releasing antibiotic resistance genes into the environment.