Team:RHS-Calgary/Safety
Safety
general safety protocols
While completing experiments, laboratory safety is always our number one priority. We work in a level one biosafety workspace. In addition to this, our entire team was required to take a written Workplace Hazardous Materials Information System (WHMIS) test prior to being allowed to work in the lab. We were required to learn about hazard classification, cautionary labelling of containers, and about how material safety data sheets are used. Though our experiments so far did not involve any biologically active substances, we have ensured to use proper sterilization procedures in the lab; For instance all pipette tips were dispelled into proper biohazard disposal containers, which were then cleaned out by a certified biohazard waste removal company that our high school has a signed contract with. We also ensure to spray down all materials with a 50% solution of ethanol, to keep everything as sterile as possible between transportation of materials. Experiments were also placed in the fume hood and all materials were properly removed in order to avoid younger classes in our school from being able to access our experiments.
lab risks
1. Which whole organisms, including viruses and cell lines, are you planning to use (or using) in your project?
Bacteria Species: Escheria coli BL21 (DE3), B. subtilis and DH5-alpha strain
Enzyme Species: Glucose Oxidase-Histag Aspergillus niger optimized for B. subtilis (BBa_K2934000) or Glucose Oxidase - A. niger (BBa_K2238000). (or BBa_K2238002)
2. What risks could these organisms pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab?
EE. coli K12-derived strains including BL21 (DE3), DH5alpha, and B. subtilis do not pose risk (cause
disease) to healthy adult humans as they lack the machinery required to colonize the digestive
system (Risk Group 1). In addition, E. coli K12-derived strains survive poorly in the environment
and are not known to cause any harm to other microorganisms or plants.
3. What organisms are you using as chasses in your project?
Escheria coli BL21 (DE3) and DH5-alpha strains will be used as a chassis in our project.
4. What risks could your chassis pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab?
The strain of E. coli that we are planning to use in our project, Escherichia coli BL21 are known to not cause harm to other microorganisms or plants and cannot colonize the digestive system of healthy adults. The risk to the community and environment is very low.
5. What experiments will you do with your organisms and parts?
Our chasses (Escheria coli BL21) will be used to create our enzyme (Glucose Oxidase). Glucose Oxidase catalyzes a reaction that converts glucose into D-glucono-1,5-lactone and hydrogen peroxide. Coupled with this reaction, we will use a colorimetric peroxide assay which will convert xylenol orange from orange to purple in the presence of hydrogen peroxide. Xylenol orange only expresses a colour change in the presence of hydrogen peroxide, so it will not react with other compounds present in feline urine. In early stages our experiments will use Thermo Scientific’s Pierce Quantitative Peroxide Assay Kit until we develop our own colorimetric assay. For our experiments we will test the reactions with gradient concentrations of glucose in a glucose solution, synthetic urine, feline urine and then feline urine in different types of litter. All of these experiments will be done in our lab or in a collaborator’s lab at the University of Calgary.
6. What risks could arise from these experiments?
The experiments do not require the use of any sharp materials and none of the compounds used pose any significant health risk. We will be using glassware, corrosive reagents, and occasional bunsen burners during our experiments.
7. How will experts overseeing your project help to manage any of the risks you identified in this form?
We are directly supervised by Dr. Iaci Soares, who has extensive experience with biological research, particularly she has a PhD in biochemical and medical pharmacology. She has studied in areas of neuroscience and cancer where she worked with similar bacterial lineages, living cells, immortalized cells, and primary culture from mice brains. Specifically, she has worked in labs with a level three biosafety level, with biohazard materials such as gmo bacteria, human cells, mice cells. In addition, Dr. Soares is a member of the school’s occupational health and safety committee and supervises all activities in the lab. The occupational health and safety committee inspects the laboratory regularly and is promptly available should there be any accidents.
8. How will the rules, training, containment and other procedures and practices help to manage identified risks?
Our laboratory space and equipment are always sanitized prior to and after use. Proper waste management and disposal is always consistently upheld, in order to avoid contamination and having any of our organisms escape the lab). While conducting experiments lab coats and gloves are always worn, as well as masks. Additional PPE was worn as necessary. Our lab is equipped to deal with all of these potential risks including an eyewash station, first aid kit, fire extinguisher, fume hood and regulated storage of all chemicals, with the safety data sheet describing the hazards and safety procedure for contact with various compounds. Experiments are always supervised by a mentor that is knowledgeable in lab protocols and who understands the details of our project. All team members and mentors have received safety training and our school has a qualified health and safety committee, should there be any laboratory accidents.