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Before beginning work on the project, every lab member underwent training with the Advisor on various safety protocols and proper techniques to be followed in lab, including how to properly handle biological organisms and parts, how to use equipment safely, measures to keep the lab area clean and precautions when using carcinogenic materials such as ethidium bromide. Students were advised to wear long pants, close-toes shoes, and appropriate personal protective equipment. This training followed the requirements outlined in the laboratory safety training requirement of UT Dallas, which can be found at http://www.utdallas.edu/research/orc/lab_safety_training.
E. coli strain DH5α was acquired from NEB and used as a chassis throughout the project. The species falls under Risk Group 1 as classified by the American Biological Safety Association (ABSA) and can potentially cause skin and eye irritation in humans. Had our project progressed to further stages, the use of Vibrio cholerae would have been required, which is in Risk Group 2 and poses greater risk to humans, requiring more precaution in handling it.
Local Rules and Regulations
Who is responsible for biological safety at your institution?
Sanaz Okhovat and Shane Solis at the Office of Research Compliance are in charge of biology safety at UTD.
What are the biosafety guidelines of your institution? Please give a link to these guidelines.
http://www.utdallas.edu/ehs/manuals/docs/BioSafety%20Manual.pdf
In your country, what are the regulations that govern biosafety in research laboratories? Please give a link to these regulations.
http://www.cdc.gov/biosafety/publications/bmbl5/bmbl.pdf
Risks of the Project
Risks to the safety and health of team members, or other people working in the lab:
E. coli is a frequently used organism on lab, which carries risk of lab-acquired infection since there are inherent risks of the organism spreading in the lab if it is not properly handled and contamination occurs.
Risks to the safety and health of the general public (if any biological materials escaped from your lab):
In the worst-case scenario, an antibiotic-resistant form of E. coli escapes lab and ends up the water source, where the likelihood of infection increases significantly and the bacteria is not responsive to the antibiotics administered for treatment.
Risks to the environment (from waste disposal, or from materials escaping from your lab):
If the organisms are not disposed of properly and end up outside of the work environment, the E. coli may end up in the water sources and affect the local ecosystems. In such a case, the organisms may interact with the bacteria present in the sewer systems and affect the native bacterial population.
Risks to security through malicious mis-use by individuals, groups, or countries:
In the hands of someone with malicious intent, the bacteria may be purposefully released into the environment. To minimize this risk, all experiments will be carried out in an area that is secured against unauthorized entry. The building in which our lab is located is also under constant surveillance.
What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)
The project, which targets various cholera genes, will be carried out in E. coli and the use of V. cholerae will be extremely limited, if at all ordered. We will take caution in handling organisms by limiting movement to the area and use personal protection equipment such as gloves and lab coats. Actions such as frequently changing gloves and cleaning the workbench with alcohol will be frequently implemented. By using efficient lab techniques and safe practices, we will keep the lab and the students working in it as safe as possible.
What new risks might arise from your project's growth? Also, what risks might arise if the knowledge you generate or the methods you develop became widely available?
We will demonstrate our model in E. coli, but our ideal final product would be a common probiotic bacterium, which poses very little risk to the lab workers, the public, or the environment. Ideally, our end product would contain a lytic phage specific for Vibrio, creating very little risk to the public and natural gut flora. It is possible for someone with ill intentions to alter our system and cause harm.
Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks?
Yes, one aspect of the project will involve using a lytic phage under an inducible promoter. This would kill our engineered E. coli.
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