Team:IISER Kolkata/Safety
Safety
Biosafety
Biosafety and Biosecurity at Research Laboratories in India is governed by the Department of Biotechnology (DBT), Govt. of India. The detailed rules and regulations can be found here. Team IISER Kolkata has abided by all the safety and security rules mentioned therein.
The team strictly abided by the safety regulations developed by the Institutional Biosafety Committee (IBSC) of IISER Kolkata,that ensures a safe workplace, acknowledges the possibility of accidents, and realizes the loss prevention benefits of safety management. The program is designed to produce a safe working environment and to eliminate or reduce hazards to employees, students, visitors, and the environment.
Our laboratory supervisors were responsible for the training of all the team members in safe practices, for correcting work errors and conditions that may result in personal injury, including exposure to carcinogenic chemicals, and for developing a positive attitude toward safety in laboratory operations. Prior to using any equipment we were briefed on the safe usage procedures. Additionally we were shown the locations and usage instructions of the fire and chemical extinguishers; emergency showers and eyewash stations.
General laboratory safety
- Protective clothing - Correct laboratory attire has been consistently enforced, including lab coats, safety glasses and nitrile gloves, as well as closed toed shoes and appropriately tied back long hair. Disposable gloves were discarded after each use, immediately after known contact with a chemical carcinogen, and upon becoming visibly wet with a chemical.
- Eating, drinking and smoking - Eating, drinking, smoking, chewing gum or tobacco, application of cosmetics, or storage of food in laboratory areas were strictly prohibited.
- Cleanliness - All team members were responsible for keeping the lab clean and clutter-free at all times. The workspace and the lab equipment were thoroughly sterilized before and after every experimental procedure.
- Waste disposal - Disposal of contaminated waste into designated biohazard waste material bins to ensure no accidental release of our modified strains into the environment, and proper disposal of hazardous and carcinogenic substances (like used agarose gels containing Ethidium Bromide). Bacterial culture containers were properly cleaned with bleaching powder followed by sterilization and autoclave.
- UV transilluminator - A Ultraviolet (UV) transilluminator with a hinged shield was used to cut gel pieces after electrophoresis. Care was taken to ensure the use of full-sleeved lab coat, safety goggles and gloves were worn while operating the instrument to prevent exposure to UV radiations.
Living organisms
- Bacterial chassis : Escherichia coli DH5 - alpha
For the genetic manipulations we worked with Escherichia coli DH5 - alpha chassis, which is non parasitic, non-pathogenic, belongs to Risk Group 1 (low risk) and can be handled at Biosafety Level 1 (BSL-1) and pose little hazard risk to laboratory personnel as it is not known to consistently cause disease in healthy adults. All experimental procedures involving the bacteria were carried out in a laminar flow cabinet shown in the image below:
Working with genetically modified organism (GMO) involves risks, such as the unintentional release of GMOs in non-GM facilities leading to potential impacts on non-target organisms, and horizontal gene transfer of genetic material. To avoid this we maintained standard biohazard protocols to discard live organisms. All the containers were cleaned with bleach followed by ethanol and finally autoclaved.
- Leishmania major
Our project aims to combat the deadly disease Leishmaniasis aka Kala Azar which is caused by genus Leishmania. For all the experimental purposes, we used less virulent form Leishmania major, which causes Cutaneous Leishmaniasis. Class II Type A biological safety cabinet was used when working with Leishmania major. Outside personnel, or those with an increased risk of contamination, were restricted from entering when work was being conducted. The team members working with Leishmania major received specific training related to the handling of the protozoa. All the experiments were performed in Dr. Rupak Datta’s lab under the supervision of his lab members who are well experienced in working with the pathogen.
- Macrophages
For all the experimental purposes Murine Murine Macrophage J774A.1cell line was obtained from ATCC. The cell line is known to be free from any kind of pathogen. The cell line is derived from BALB/c mouse and not known to cause disease. The experiments were performed under the direct supervision of lab instructors inside Class II Type A biological safety cabinet. We abided by all the standard biohazard procedures while discarding the live cells.
Management of Chemical Hazards
- Ethidium Bromide safety
Ethidium bromide (EtBr) is commonly used as a non-radioactive marker for identifying and visualizing nucleic acid bands in electrophoresis. Although it is an effective tool, its hazardous properties require special safe handling and disposal procedures. In addition to wearing proper lab attire and designated gloves within EtBr areas, while working with or around EtBr we only used designated lab equipment and instrumentation. Any waste connected to EtBr was disposed off in designated EtBr waste containers.
- Chemical Fume Hood
A fume hood is a type of local ventilation device that is designed to limit exposure to hazardous or toxic fumes, vapors or dusts. While dealing with hazardous chemicals, we used a chemical fume hood.
- Glovebox
A glovebox is a sealed container that is designed to allow one to manipulate objects where a separate atmosphere is desired. The experiments involving DETA NONOate, which is a Nitric Oxide donor, were carried out in a glovebox to achieve an inert environment.
