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Part I Guidelines For Safe Working Practices In the GSBmE PC2 Laboratory Version 3, 2009 Lynn Ferris Mai Ly GSBmE University of NSW Office of the Gene Technology Regulator Part II Introduction What is a GMO General practice Dry work Liquids Biological Safety Cabinets Spills Cultures
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Lynn Ferris
Mai Ly
GSBmE University of NSW
Office of the Gene Technology Regulator
Part II
What is a GMO
General practice
Dry work
Liquids
Biological Safety Cabinets
Spills
Cultures
Storage
Transport
Waste
OHS
Equipment
Further information
Part I: Safe Working Practices in the GSBmE PC2 LaboratoryIndexPart I: Guidelines for Safe Working Practices in the GSBME PC2 Laboratory
Part II: Behavioural Requirements for working in the OGTR PC2 Laboratory at GSBME
Eat, Drink or Smoke in the facility
Apply cosmetics
Mouth pipette
Insert contact lenses
Bring or store food
Tongue moisten labels
Contaminate materials (e.g. workbooks) that will be removed from the room without sterilization.
Keep hands and pens away from your face. They may have been in contact with contaminated surfaces or aerosols
Tie back long hair
Windows must be kept shut.
You must wear closed footwear
Avoid using bleach or other chlorinated chemical disinfectants
You must notify the Lab Manger of any spills or accidents immediately
The lab doors should always be kept closed
Take care that reading and writing materials do not become contaminated.
Part I: Safe Working Practices in the GSBmE PC2 LaboratoryGeneral PracticeThey are kept within the PC2 room. Remove your external coat before entering the PC2 lab. Hooks are provided outside the room.
Do not bring personal items such as mobile phones or backpacks into the PC2 lab
Wear gloves
All gloves should be disposed of in the biohazard waste
Wipe down ALL work surfaces before and after use with the 80% (v/v) ethanol sprays provided
Paper tissues are provided as wipes and should be disposed of in the biohazard waste
Remove both gown and gloves just before leaving the room.
Always wash your hands with the disinfectant provided before you exit the PC2 room
Part I: Safe Working Practices in the GSBmE PC2 Laboratory Dry WorkAerosols can be produced during
Vortexing
Sonicating
Centrifugation
Pipetting
Opening containers
Minimise aerosol production on open benches. Use closed containers for shaking and mixing
Use the biological safety cabinets where possible when you are opening, mixing, decanting or aliquoting fluids.
Part I: Safe Working Practices in the GSBmE PC2 Laboratory LiquidsDecontaminate the hood with UV light for a minimum of 20 minutes
Organise all your materials for your session so that they are easily available.
After use wipe down surfaces (80% (v/v) ethanol) and decontaminate with UV light for 20 minutes
The biological safety cabinets are NOT fume hoods - They will not protect you from harmful chemical fumes. Fume hoods are located in rooms 429, 430 and 431
GSBME has safe-work instructions posted on each Cabinet.
Part I: Safe Working Practices in the GSBmE PC2 LaboratoryBiological Safety CabinetsPrior to work in biological safety cabinets, you should:
If infectious material is spilt, avoid breathing in any aerosol... wait 5-10 minutes until the particles have had a chance to settle
Cover the spill gently with adsorbent towel, sand or vermiculite and dispose in the contaminated waste
Wipe down the area and equipment with 80% (v/v) ethanol
Report all spills of GMO’s to the Area Supervisor or the Lab Manager.
Part I: Safe Working Practices in the GSBmE PC2 Laboratory SpillsBacterial cultures should be cultured and incubated separately from the mammalian cell lines
All cultures should be stored within the designated areas
All cultures (including GMOs) must be labelled with:- date- identification- owner
Any unlabelled viable materials must be treated as a GMO and handled in accordance these conditions
Notify the Lab Manager immediately
Part I: Safe Working Practices in the GSBmE PC2 Laboratory CulturesAll genetically modified organisms (GMO) will be stored in a designated biohazard containment area
The register of GMOs will contain storage location details and other identifying information
All work with GMOs must be approved by the UNSW Institutional Biosafety Committee.
Part I: Safe Working Practices in the GSBmE PC2 Laboratory StorageFor GMOs, the secondary container must be unbreakable.
The outside container must be labelled with a name and contact details in case the package is lost
For GMOs, indicate that it is a GMO
Both containers must be decontaminated or disposed as a biohazard waste after use to ensure no residue remains
Organisms transported by road, rail or air must comply with transport regulations
Any GMO samples or cultures taken from the PC2 laboratory shall go only to another PC2 containment facility
Part I: Safe Working Practices in the GSBmE PC2 Laboratory TransportWhen the drum is full, the top of the bag is folded inside the drum and the lid placed on top
The whole drum is then placed in the autoclave without the lid to allow steam penetration
500ml of water is poured down the side (inside the waste bag) to facilitate steam generation
Heat sensitive tape must be applied to the biohazard bag to differentiate from unautoclaved waste. You must be trained before using the autoclave
The autoclave waste is treated at 121°C for a minimum of 20 minutes (BE S006). Ensure that the entire contents are subjected to these minimum conditions. A full drum normally requires 60 minutes of treatment.
An indicator is used once a month to validate successful sterilisation of waste
After treatment, the bag is removed from the drum and placed inside a second biohazard bag, sealed with string and labelled with a red biowaste sticker
Part I: Safe Working Practices in the GSBmE PC2 LaboratoryWaste Disposal“Tissue culture waste”
Name
Weight
“Biomedical Engineering, UNSW”
Date
The sterilised waste is then placed in the contaminated waste collection bin in the cool room (405)
Liquid waste
Liquid waste is treated with a chemical disinfectant eg 1% w/v available iodine solution
Avoid using bleach or other chlorinated disinfectants as they may release harmful gases
The treated liquid is then placed in the biological liquid waste container in the cool room (405)
Part I: Safe Working Practices in the GSBmE PC2 LaboratoryWaste DisposalContAll waste is removed by the UNSW contractor
When flame sterilising, have a cover ready to smother the flame
Aliquot minimal amounts of ethanol
Ensure that flame sterilised tools are long enough to hold safely
Hold flame tools horizontally to prevent heat and flame transfer to your hands
Part I: Safe Working Practices in the GSBmE PC2 Laboratory Occupational Health and SafetyToxic chlorine gas can be released from chlorine when it is heated
Iodine based disinfectants should be used instead at a 1%w/v available iodine minimum
All injuries, spills or near misses should be immediately reported to the laboratory manager
Immunisation eg hepatitis B may be recommended for particular workers.
Part I: Safe Working Practices in the GSBmE PC2 Laboratory Occupational Health and Safety ContThe use of the autoclave is not permitted without proper training and authorisation
For training see Veronika Tatarinoff room 434 ext 53923
ALL work must be covered by an authorised Risk Assessment.
GMO work protocols must comply with statutory regulations
All chemicals and reagents must be appropriately assessed and labelled according to the Hazardous Substances Regulations
Use the GSBmE standard label on your preparations and quote your relevant Risk Assessment number
Part I: Safe Working Practices in the GSBmE PC2 Laboratory Occupational Health and Safety ContNo equipment is to be removed from the PC2 facility without the approval of the Area Supervisor or Lab Manager
All equipment must be decontaminated before transfer outside the PC2 facility.
All equipment must be decontaminated before repairs are made.
All equipment must be cleaned and maintained. Records of maintenance must updated.
Maintenance Contractors must report to the Area Supervisor or Lab Manager prior to starting work.
Report any equipment or contractor issues to the Area Supervisor or Lab Manager
Part I: Safe Working Practices in the GSBmE PC2 Laboratory EquipmentSOP BE L031-01Routine Cleaning of Equipment used in a PC2 Laboratory
SOP BE S006-01
Overview of Sterilisation Techniques
Autoclave Training
Fundamental Autoclave Techniques
https://www.gsbme.unsw.edu.au/ohs/autoclave.php
Risk managementOHS Coordinator (Biosafety)Adam Janssen ext 52214 a.janssen@unsw.edu.au
UNSW risk management http://www.riskman.unsw.edu.au
Office of the gene technology regulatorhttp://www.ogtr.gov.au
PC2 area supervisorMai Ly ext 53909
Laboratory Manager
Lynn Ferris ext 53909
Part I: Safe Working Practices in the GSBmE PC2 Laboratory Further InformationMai Ly
GSBmE University of NSW
Office of the Gene Technology Regulator
Part I
Doors & windows
Non-GMOs in the PC2 Lab
Containment equipment
Personal protective clothing
Decontamination
Labelling
Removal & storage of GMOs
Spills
Further information
Part II: Behavioural Requirements for GSBmE PC2 LaboratoryIndexPart I: Guidelines for Safe Working Practices in the GSBME PC2 Laboratory
Part II: Behavioural Requirements for working in the OGTR PC2 Laboratory at GSBME
GMOs must be made non-viable prior to disposal
Wastes containing GMOs must be decontaminated prior to disposal
Work benches, surfaces and equipment must be decontaminated at the completion of procedures involving GMOs
Personal protective clothing:
Lab gowns must be decontaminated prior to resuse if it has been contaminated or suspected to be contaminated with GMOs
Gloves are disposed of in the biohazard bin
Decontamination can be:
Autoclaving or other heat based treatment
Incineration
Chemical treatment
Part II: Behavioural Requirements for GSBmE PC2 Laboratory DecontaminationLoads must be packed and loaded to allow adequate steam penetration
The coldest part of the load must be exposed to a minimum of temperature of 121°C and 103 kPa for at least 20 minutes
Loads that have been processed can be differentiated from unprocessed loads (e.g. autoclave tape)
Refer to standard AS/NZS 2243.3:2002
Incineration must be performed in a high temperature, high efficiency incinerator
Autoclaved waste at GSBME is collected and incinerated by a contractor
Any chemical disinfectant treatment must be effective in rendering the GMO non-viable
Refer to standard AS/NZS 2243.3:2002
Hands must be washed prior to leaving the laboratory
Hand operated taps are not acceptable
Part II: Behavioural Requirements for GSBmE PC2 Laboratory Decontamination Cont- date - identification - owner
They are to be transported to another certified PC2 facility
They are transported to another location for storage
They are transported to another location to be decontaminated prior to disposal
Written permission by the OGTR or permits, for transport to another destination within Australia or to the Australian border for export
All GMOs transported out of the facility must be
Confined in a primary sealed container, within a secondary airtight an unbreakable container
The outside container must be labelled with a name and contact details, and indicate that it is a GMO
Organisms transported by road, rail or air must comply with transport regulations
Part II: Behavioural Requirements for GSBmE PC2 Laboratory Removal and storage of GMOsPrevents dissemination of GMOs
GMOs stored outside the PC2 laboratory (freezer, fridge, etc) must:
Have a biohazard symbol posted on the storage unit
Be locked when not in use, unless access is restricted to the room/area where the storage unit is located
Prevents unintentional release of GMOs into the environment
All GMOs stored outside of the facility must be
Confined in a primary sealed container and labelled.
The primary container must be stored in a secondary unbreakable container
In the case of a small storage unit such as fridge, freezer or liquid nitrogen dewar, the secondary container may be the storage unit
If the GMO is a notifiable low risk dealing (NLRD), you must obtain authorisation from the UNSW Biosafety Committee to store your GMOs outside the PC2 laboratory
Visit the OGTR website for more information on the different classes of GMOs
Part II: Behavioural Requirements for GSBmE PC2 Laboratory Removal and storage of GMOs ContGMO spills inside the PC2 laboratory must be decontaminated as soon as reasonably possible in accordance with the spills procedure
GMO spills outside the PC2 laboratory must be decontaminated and reported to the Lab Manager immediately.
Any real or suspected unintentional release of GMOs outside the PC2 laboratory (including spills) must be reported to the OGTR as soon as reasonably possible
Part II: Behavioural Requirements for GSBmE PC2 Laboratory SpillsGMO spills are reported to:
Lab Manager
UNSW Institutional Biosafety
Committee
OGTR
SOP BE L031-01Routine Cleaning of Equipment used in a PC2 Laboratory
SOP BE S006-01
Overview of Sterilisation Techniques
Autoclave Training
Fundamental Autoclave Techniques
https://www.gsbme.unsw.edu.au/ohs/autoclave.php
SWP
Safe Blood Work Practices
Blood Work Training
Guidelines for Safe Blood Work Practices
Risk managementOHS Coordinator (Biosafety)Adam Janssen ext 52214 a.janssen@unsw.edu.au
UNSW risk management http://www.riskman.unsw.edu.au
Office of the gene technology regulatorhttp://www.ogtr.gov.au
Australian standards
AS/NZS 2243.3:2002
PC2 area supervisorMai Ly ext 53909
Lab Manager
Lynn Ferris ext 53909
Part II: Behavioural Requirements for GSBmE PC2 Laboratory Further InformationPart I
Part II