Working in PC-1 and PC-2 Tissue Culture Facilities Description: This procedure is applied when working in PC-1 and PC-2 tissue culture labs

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1 Faculty/School: Faculty of Pharmacy Initial Issue Sep 2009 Next Review Oct 2017 SWP Reference Number: Version: 4 Version Issue Oct 2016 SWP Title: Prepared by: Working in and Tissue Culture Facilities Description: This procedure is applied when working in and tissue culture labs Dr Sarah Cui Responsible supervisor/s: Research Supervisors and Laboratory Managers List the Hazards and risk controls as per risk assessment Associated assessment reference: risk Hazards Risk controls 1. infection of worker, other people, plants or environment 2. The hazards will depend upon the individual organism, the route of exposure and the number of organisms involved Personal protective equipment, good risk assessment and management should be applied List resources required including personal protective clothing, chemicals and equipment needed PPE: a). gloves, nitrile or latex; b). lab gown (back fastening) with full-length sleeves; c). Safety glasses or goggles if splash/squirt/spray risk present; d). Hair tied back if long; e). Proper enclosed footwear; f). P2-type respiration if aerosol hazard of risk group 2 microbes exists. List step by step instructions or order for undertaking the task 1. All laboratory members working with PC1/ PC2 microorganisms should read and understand legislations and documentations listed below. 2. Lab members work with GMOs/OGTR regulated materials are required to obtain approval from university biosafety unit, and work in GMO/ OGTR certified facility; relevant training is required, contact Sarah Cui for details. 3. All laboratory members are required to read SWPs and become competent on operating a Biosafety II Cabinet, Biohazard Spill, Autoclave Operations and other procedures which are listed in the training form. 4. Users are required to complete Faculty WHS essential procedures, and send a training request to pharmacy.labsupport@sydney.edu.au for lab induction, lab access will be approved for authorised users. 5. All lab members need to understand specific hazards associated with the biological samples they are using before they start work. 6. All laboratory members must understand the specific risks associated with the organisms they work, SWP and risk assessment should be completed by the project supervisor, and new users should complete training before starting to work independently. 7. All laboratory members working PC1/PC2 labs must be trained and competent in the clean-up /decontamination of work surfaces and spills involving these biohazards, this may include risk group 1,2 microorganism. 8. If untrained (eg. undergraduate students), the lab member should be closely & constantly supervised by someone with microbiological training.

2 9. Know the location of spill kits, eyewashes, safety shower, fire extinguishers and fire blankets before starting work 10. Lab members must be aware of any pre-existing relevant health conditions, and inform their supervisor of these, if these would impact on microbiological work, eg. Immunocompromised status due to HIV infection or medication would make the worker much more susceptible to infection. 11. Never eat, drink, smoke or apply cosmetics in the microbiology lab. Never put anything in your mouth (eg. don t chew the end of pens). 12. Do not use your mobile phone while doing microbiological lab work- this can act as a vector for carrying microbes outside the lab 13. Work bench should be clear of clutter and personal possessions (eg. no bags, clothing etc). Only the equipment necessary for the tasks to be performed should be present on the bench. Do not sit on the lab bench. 14. Bench surface should be disinfected before beginning work, eg. with 80% (v/v) ethanol solution. It is good practice to sterilise any tools required. 15. All other equipment and media must be sterilised by autoclaving or filtration. 16. Work in a biosafety cabinet for any manipulations of risk group 2 organisms that are likely to generate aerosols this includes vortexing, sonication, pipetting, and opening tubes. See SWP for biosafety cabinet. 17. All cultures must be labeled with the name of the organism (species and strain), the name or initials of the worker, and the date. Cultures must be incubated in the appropriate location, and must not be taken out of the lab. 18. Risk group 2 cultures must be transported inside a secondary unbreakable container if they need to be transported outside the PC2 lab. A strong plastic box with a lockable lid is ideal with labels as in Appendix 1. See also SWP for Transport of GMOs. 19. Risk group 2 cultures must be stored in a secondary unbreakable container, and be properly labeled (as above), and records must be kept of the cultures. Also see SWP for storage of GMOs. 20. Upon finishing work, sterilise the bench again, and discard all unwanted cultures into the appropriate location. All microbial cultures must be destroyed by autoclaving; they cannot be put down the sink, or in the regular garbage. See also the SWPs for Hazardous Waste Disposal and SWP for Disposal of GMO contaminated waste. 21. Wash your hands with antiseptic soap before leaving the laboratory, eg. with soap containing chlorhexidine. In lab, lab members are required to follow the instruction on how to use hand-free hand wash facility, basically, use elbow but not hands to turn on and off the handle. Remove your lab coat or gown before leaving the laboratory. Lab coats or gowns used in tissue culture labs must not leave the facility, unless they are autoclaved first. Lab gowns are required to hang up on the hooks with sleeves folded to the inside to avoid cross contamination. lab gowns are required to be autoclaved before laundering. List emergency shutdown procedures In the event of emergency shutdown, keep minimal exposure of biohazard/gmos samples, and leave the room immediately. List Emergency procedures for how to deal with fires, spills or exposure to hazardous substances In the event of a fire emergency evaluation, leave a notice for unattended procedure keep minimal exposure of biohazard/gmos samples and leave the room. List Clean up and waste disposal requirements 1. All biohazard spills must be cleaned up immediately (see appropriate Biohazard Spill SWP). If you need to use bleach to do this, ensure you follow up with a water-soaked wipe such that the bleach residues are not left in the cabinet, these will corrode the surfaces. 2. In the case of a large spill of a risk group 2 organism (eg. >100 ml), allow 20 min for aerosols to settle before attempting to clean up the spill. Alternatively, wear a full-face respirator (located in administration office) if you need to clean it immediately. 3. This bag containing the spill materials should NOT be autoclaved, as bleach will corrode the autoclave. Instead, this should be disposed of as medical waste, or as hazardous chemical waste. Page 2

3 4. If you are exposed to infectious materials or if you become sick and you suspect this is due to biological agents you have handled at work, you must report this to your supervisor as soon as possible, and fill in an online incident report form 5. Any injuries or incidents must be reported immediately to your supervisor and within 24 h using the online incident report form. Near misses (hazardous situations not leading to an incident) should also be reported List references used in the development of this SWP, e.g. codes of practice 1. University WHS website: sydney.edu.au/whs 2. AS/NZS :2005-Safety in Laboratories: Planning and Operational Aspects 3. AS/NZS :2010-Safety in Laboratories: Microbiological safety and containment 4. AS/NZS :2004-Safety in Laboratories: Storage of Chemicals 5. AS/NZS :2010-Biological safety cabinets Classes I & II-Installation and Use 6. OGTR Guidelines for Certification of a Physical Containment Level 2 Laboratory (v.3.1) 7. OGTR guidelines for the Transport, Storage and Disposal of GMOs (v.1.1) 8. Risk assessment for biohazard spills 9. Risk assessment and SOP for Risk group 2 microorganisms and / or Animals + Animal Tissues and / or Humans/ human tissues List competency required qualifications, certificates, licensing, training - e.g. course or instruction: The staff and students, who have completed onsite training by technical officers or Lab Manager could obtain swipe card access authorisation to the research area, after project-specific training by supervisors, they could work in the laboratory. All the lab members should attend Biosafety training organized by university of Sydney WHS unit, and members handling GMO/OGTR are required to complete university Biosafety OGTR training, Staff approved to assess competence for this SWP Technical Officers and Lab Manager Page 3

4 SWP Sign off sheet SWP name and version: In signing this section the assessor agrees that the following persons are competent in following this SWP. Name Signature Date Competent Name Assessor/Authoriser of Assessor/Authoriser signature Appendix 1. Labels for transportation of biological samples Page 4

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