Team:DeNovocastrians/Safety




Safety


Lab Safety

The University of Newcastle is an accredited organisation with the Office of the Gene Technology Regulator (OGTR) and subsequently must adhere to biosafety and security regulations to maintain accreditation. The DeNovocastrians team fully complied to the strict rules and regulations put in place by the University of Newcastle and OGTR. Lab work was conducted in a PC2 laboratory following strict guidelines set out by the OGTR. These regulations cover lab-based work or dealings with genetically modified organisms, and also outline requirements for physical containment (PC) levels of labs when dealing with various microorganisms.

By adherence to standard PC2 laboratory practices, potential risk is greatly reduced. Practices including wearing appropriate personal protection equipment, following correct chemical disposal and handling reduces the risk of any spillages or personal harms. The laboratories were under supervision of qualified academics and were inspected regularly by the safety committee to ensure the laboratories were up to standard, and everything was done according to legislation.

The team's PIs and advisors supervising lab work carried out by the undergraduates have extensive experience working in microbiology labs. They are familiar with the experimental procedures that were used, such a microbial culturing, cloning, PCR, and correct handling and disposal of hazardous substances.



Handling Microorganisms

We had access to two species of Rhodococcus strain 9 and Rhodococcus strain 33, both isolated from benzene contaminated systems in Australia, there is no evidence for the pathogenic potential of these strains. Since both strains are naturally found in soil and aquatic habitats, they pose minimal risk to the environment were they to be inadvertently released from the lab. Gene clusters from Acinetobacter and Escherichia fergusonii were synthesized from biobricks, cloned, and handled with care.

While exposure to any microorganism in a high density culture can be considered a risk, Escherichia coli (BW25113/BL21/DH5alpha), ADP1, Synechocystis and Bacillus subtilis (168) lab strains are generally considered non-pathogenic and pose minimal health risk within or outside of the laboratory. They are White list or Risk Group 1 microorganisms and therefore present minimal environmental risk should they be inadvertently released from the lab.

All work involving live microorganisms was conducted in a PC2 lab and organisms were disposed of in accordance with containment procedures, therefore the risk of release outside of the lab is extremely low. Even so, all necessary precautions, such as correct PPE use and handling safety, were taken when using them.



Material Substitutions

Benzene and catechol are mutagens and carcinogens that require special handling and disposal. The benzene and catechol work was approved by the University of Newcastle Health and Safety department. To reduce risk, team PIs and advisors (PhD students) carried out and instructed on correct management, handling, and disposal of hazardous materials, including chemicals and biological materials. To reduce the risks of working with benzene, we worked with the less toxic chemical benzoate instead of benzene. Team members were correctly trained in the safe handling and disposal of benzoate. Those working directly with benzoate underwent a health surveillance procedure. An application outlining the hazards and risk management for this work was also submitted to the University of Newcastle's Institutional Biosafety Committee for approval.

DNA gels were stained with ethidium bromide (a known mutagen) for imaging, these tasks were only performed by PIs and advisors.



Biosensor

We also developed a rapid detection biosensor to detect levels of benzoate and catechol in environmental systems using Green fluorescent protein and mCherry. Our biosensor will not be implemented as it would require extensive testing to clear for use outside of the lab, for which we did not secure permits.