Team:Exeter/Safety
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Safety
INTRO
researchers and the public need to be kept safe when biologically engineering organisms
Safe Project Design
Decisions made to reduce risk of design
We used E. coli strains, such as TOP10 and DH5α, which are non-pathogenic chassis for cloning, typically used in labs. [1]
We removed the manose binding region from the pili. ****IS THIS RIGHT??**** This binding is a natural virulence factor in pathogenic strains of E. coli. This was in effort to choose a part and reduce its harm to humans, animals or plants and repurpose it for metal ion binding.
We have substituted the manose binding domain for metallothienein domains in a proof-of-concept experiment for a commercial product. Metallothienein is less dangerous than the natural manose binding domain.
Protocols have cautious safety measures incorporated in preventing the spread of any living organisms we used.
Filter design incorporates a UV light ***IS THIS RIGHT**? to prevent organisms entering the environment
Safe Lab Work
Not all the team members had experience in labs before so all wet lab work was supervised by at least one member of staff.
‘Boot camp’ safety training week incorporated lessons on:
* Aseptic technique
* COSHH forms
* cleaning surfaces with 70% ethanol
* emergency protocols
* Lab introduction
* Location of fire exits
* First aid points
* Handling waste
* Handling toxic chemicals
* Using the designated area for running gels
Safety procedures used every day in the lab
We worked in a wet lab safety level 2 that covers work with agents associated with human disease, pathogenic or infectious organisms posing a moderate hazard. Precautions taken were:
* contaminated sharps are handled with extreme caution
* biosafety cabinet
* autoclave
* eyewash station
* self closing lockable doors
* biohazard warning signs at all access points
* flow hoods used
* glassware sterilization using virkon
Daily precautions involved:
* Personal Protective Equipment
* Lab coats
* Blue nitrile gloves
* Long hair tied back
* Enclosed toe shoes
* Mask and goggles when dealing with sans sifting
* Use flow hoods where necessary
* Appropriate storage of different chemicals
* Waste disposal such as discarding waste tips, petri dishes and falcon tubes
* Autoclaving methods
* No food or drink allowed in the lab
* Aseptic technique
* (biosafety) reduced contact with biological material
* Clean and organised work bench
* Hand washing after leaving the lab
Unusual experiments/unusual safety issues?
We visited the Wheal maid site which is a mine waste water site and collected samples from the contaminated site. Various precautions were taken:
* wore nitrile gloves, waterproof jackets and wellies
* filtered and sealed water samples in sample bottles using sterile syringes
* treated and prepared samples for metal ion composition analysis
* dealt with HCl and HNO3
* handled metal ion standard stock solutions
Safe Shipment
Did you face any safety problems in sending your DNA parts to the Registry?
****NOT DONE YET******
How did you solve those problems?
****NOT DONE YET******
Application safety issues
If our genetically modified E. coli was used in the final filter product it could create some safety issues. E. coli is therefore only being used as a chassi and as a proof of concept.
Further studies should be conducted to apply our concept to one of the naturally occurring organisms found thriving in mine waste water habitat conditions.
There are likely to be concerns from the general public as to the potential chance of mutation and escape of these bacteria into the local waterways. To reduce the potential of this scenario occurring, there are stages within our filter system that will destroy any bacteria entering and leaving the filter we planned to use _____******_____****_______
UNANSWERED QUESTIONS
Biosecurity - how will we prevent our product being used for harm
COSHH forms
Lab
* Viewing gels on uv transilluminator
* Midori green , SYBR safe stain
* Metal ion concentrations
* Centrifuge
* geneJET plasmid miniprep kit
* preparing LB medium & pouring plates
* agarose gel electrophoresis of nucleic acids
* making antibiotic solution
Field trip
* Travelling there
* Sampling at the site
* lab prep of samples using acids
Standard Operating Procedures for sample analysis using the ICP-OES
Risk assessment for diluting acids used in SOP
Risk assessment for handling metal ion standard solution used in SOP
Filter
* Sand sieving – wore facemasks and goggles
* 4% Mannose
Transport
* travelling
References
Potts, C. (2015). Statement of nonpathogenicity. [PDF]. Available at: https://www.thermofisher.com/content/dam/LifeTech/global/technical-reference-library/cloning/pdfs/statement-of-nonpathogenicity.pdf
