Our lab at TAS is classified as Biosafety Level 1, which means the lab uses microbes that pose little to no risk to healthy individuals. Hazardous chemicals and solutions are used at a minimal level and in well ventilated areas such as fume hoods in the TAS research lab. For instance, we do not use EtBr for gel electrophoresis. Instead, we use a safer nucleic acid stain called Seeing Safe DNA Dye. Nevertheless, all chemicals and solutions are still treated with all due respect, care, and caution. MSDS are stored on our lab computers and in a folder on the lab technician's desk.

We work with a safe and common lab strain, E. coli K-12 (DH5alpha), and follow safety rules set by our lab instructor. We have our own biosafety committee, which consists of three research teachers Dr. Teresa Chiang, Mr. Jude Clapper, and Mr. Alex Dezieck. They oversee proper work area conditions by checking on disposal of Petri dishes and liquid wastes, sanitation, and teaching proper laboratory techniques. Our guidelines, taken from National Yang-Ming University's Center of Environmental Protection and Safety, cover many safety rules and procedures ranging from lab specific rules to behaviour. For example, we prohibit food, open-toed shoes and drinks in the lab. We also have a thorough clean up procedure. For example, we have waste bins for used tips, which are autoclaved before disposal, and liquid wastes are bleached. Nanoparticle (NP) waste is carefully packaged and then sent to a disposal facility to ensure no hazardous materials are mismanaged.

In Taiwan, there are many laws and regulations regarding biosafety in research labs. These can be found in the National Yang-Ming University's Center of Environmental Protection and Safety and Health page linked here. Guidelines can be found here.

PROJECT-SPECIFIC BIOSAFETY

Nanoparticles

Upon seeing the increased usage of NPs in medical, industrial, and consumer products, our team looked into rules and regulations regarding disposal of NPs. Since we were going to be using NPs in our research, we needed to make sure that we were disposing our waste properly. We discovered there are few regulations on NPs, but we found a local company (THEPS LIMITED) to pick up our own NP waste products, which are then transported to Cheng Gong University for incineration.

Prototype

We have chosen to use a safe and common lab strain of E. coli, K-12, as our chassis (Environmental Protection Agency 1977). In both approaches, our constructs do not express proteins associated with virulence. PR is a membrane protein that commonly exists in marine bacteria. As for biofilm production, we were careful to avoid known virulence factors such as alpha hemolysins (Fattahi et al. 2015). However, biofilms are sometimes associated with health problems, so we plan to only use our biofilm in wastewater treatment plants (WWTPs). Biosafety is built into WWTPs; before treated effluent is released back into the environment, it must go through a final disinfection step, where chlorine, ozone, or UV radiation are used to kill microbes still present in the wastewater (Pescod 1992).

REFERENCES

Environmental Protection Agency. (1977). Final Risk Assessment of Escherichia Coli K-12 Derivatives . EPA. Retrieved from https://www.epa.gov/sites/production/files/2015-09/documents/fra004.pdf.

Fattahi, S., Kafil, H. S., Nahai, M. R., Asgharzadeh, M., Nori, R., & Aghazadeh, M. (2015). Relationship of biofilm formation and different virulence genes in uropathogenic Escherichia coli isolates from Northwest Iran. GMS Hygiene and Infection Control, 10, Doc11. http://doi.org/10.3205/dgkh000254

Pescod, M. (1992). Wastewater treatment and use in agriculture (Vol. 47). Rome: United Nations.