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WHS UNIT

BIOSAFETY MANUAL Contents 1 Introduction ............................................................................................................................................ 3 2 Scope ..................................................................................................................................................... 3 3 Guiding Principles .................................................................................................................................. 3

3.1 Introduction ........................................................................................................................................ 3 3.2 Objectives .......................................................................................................................................... 3 3.3 Responsibilities .................................................................................................................................. 4

4 Definitions .............................................................................................................................................. 5 5 The Laboratory ...................................................................................................................................... 6

5.1 Containment Measures ...................................................................................................................... 6 5.2 Physical Containment Classifications ................................................................................................ 6 5.3 Laboratory Biosecurity ....................................................................................................................... 7

6 Administration ........................................................................................................................................ 7 6.1 Contact Details .................................................................................................................................. 7 6.2 Laboratory Access and Authorisation ................................................................................................ 7 6.3 Annual PC2 Laboratory Inspections .................................................................................................. 7

7 Training .................................................................................................................................................. 8 7.1 Local .................................................................................................................................................. 8 7.2 General Biosafety .............................................................................................................................. 8 7.3 Genetically Modified Organisms ........................................................................................................ 8 7.4 Training Details .................................................................................................................................. 8

8 Purchasing/Acquisition .......................................................................................................................... 9 9 Risk Management .................................................................................................................................. 9

9.1 Register of Hazards ........................................................................................................................... 9 9.2 Risk Assessments ............................................................................................................................. 9 9.3 Safe Work Procedures ....................................................................................................................... 9 9.4 Incident Reporting .............................................................................................................................. 9 9.5 Personal Injuries ................................................................................................................................ 9 9.6 Personal Protective Equipment (PPE) ............................................................................................. 10

10 Pest Control Program .......................................................................................................................... 10 11 Facility Work Practices ........................................................................................................................ 11

11.1 PC1 Facility ...................................................................................................................................... 11 11.2 PC2 Facility Work Practices ............................................................................................................ 11 11.3 GMO Physical Containment Facilities ............................................................................................. 12 11.4 Quarantine Approved Premises (QAP) ........................................................................................... 12

12 Biological Safety Cabinets ................................................................................................................... 12 13 Working after Hours ............................................................................................................................. 13 14 Health Management ............................................................................................................................ 13

14.1 General ............................................................................................................................................ 13 14.2 Immunisation ................................................................................................................................... 13 14.3 At Risk Persons ............................................................................................................................... 13 14.4 Precautions for Pregnant Women ................................................................................................... 13 14.5 Health monitoring ............................................................................................................................. 13

15 Personal Hygiene ................................................................................................................................ 14 16 Disinfectants ........................................................................................................................................ 14 17 Laundering Of Laboratory Gowns ....................................................................................................... 14

17.1 Safe Removal of Contaminated Gloves .......................................................................................... 14 18 Biological Waste .................................................................................................................................. 14 19 Emergency Management for Microbiological Spills ............................................................................. 15

19.1 General ............................................................................................................................................ 15 19.2 Factors Influencing Response to Spills ........................................................................................... 15

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19.3 Planning for the Management of Spills ............................................................................................ 15 19.4 Basic Biological Spill Clean-up Kit ................................................................................................... 16 19.5 Clean-up Procedures ....................................................................................................................... 16 19.6 Spills Inside Biological Safety Cabinets ........................................................................................... 16 19.7 Spills Outside Biological Safety Cabinets ........................................................................................ 17 19.8 Centrifuge Spills ............................................................................................................................... 19 19.9 Disposal of Contaminated Waste Following Spill Cleanup .............................................................. 19

20 Fieldwork.............................................................................................................................................. 19 21 References........................................................................................................................................... 19 22 Version Control Table .......................................................................................................................... 21 Appendix 1 – Biosafety Legislation and Guidance .......................................................................................... 22 Appendix 2 - Summary of Recommended Applications for Chemical Disinfectants in Microbiological Laboratories ..................................................................................................................................................... 24 Appendix 3 - Good Hand Washing Technique ................................................................................................ 25 Appendix 4 - Removing Gloves Safely ............................................................................................................ 26

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1 Introduction This manual provides guidelines on relevant legislative requirements and safe work practices when working with or being exposed to biological hazards. These guidelines should be used in conjunction with the University of Wollongong (UOW) Laboratory Safety Guidelines. Guidelines for working with biological hazards are regulated by the Office of the Gene Technology Regulator (OGTR) when working with GMOs, the Department of Agriculture and Water Resources, Biosecurity Australia (formally AQIS) when working with imported materials and the AS/NZS 2243.3:2010 when working with general microbiological organisms and diagnostic samples.

2 Scope This manual applies to general biosafety practices as well as PC1 and PC2 laboratory practices. Any PC3 laboratory practices must be specifically developed for the work being conducted in accordance with AS/NZS 2243.3:2010 and be approved by the Dean or the Director of the Unit.

3 Guiding Principles

3.1 Introduction UOW is committed to providing its staff and students with a safe working, research and study environment. This includes controlling biological hazards to ensure that safe and productive work practices are carried out, and that exposure to biological hazards is minimised. As an educational and research institution, the University seeks continual improvement in awareness and prevention of exposure to biological hazards. The University follows the relevant legislation and standards such as those prescribed by OGTR, the Department of Agriculture and Water Resources and Australian Standards.

Biological hazards that staff and students may be exposed to at UOW include:

human blood, tissue and body fluids human sewage and faeces microorganisms cell culture GMOs animals (including their tissues, blood or body fluids and excreta) contaminated soil and water samples plants.

3.2 Objectives The objectives of this document are to ensure that:

all staff and students working with biological hazards receive appropriate information and training,

enabling them to recognise the hazards and risks they may encounter during the course of their work biohazardous operations involving microorganisms or diagnostic samples are performed in the

appropriate manner and physical containment facility according to their risk group (includes storage and transportation)

work involving GMOs be approved by the UOW Gene Technology Review Committee (GTRC) and carried out under the guidelines of the OGTR

the use of humans or animals and their tissue, blood or body fluids in research and teaching receives approval from the Research Ethics and Animal Ethics Committees prior to commencement of work on these materials

emergency management procedures are in place in the event of a biological spill or release appropriate waste management systems to effectively dispose of biological waste are in place.

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3.3 Responsibilities

3.3.1 Deputy Vice Chancellor (Research)

The Deputy Vice Chancellor (Research) is responsible for ensuring that:

all work carried out using biological hazards at UOW is compliant with the relevant legislation UOW maintains the necessary systems and procedures for accreditation with the OGTR an institutional biosafety committee is established to conduct and maintain safety requirements as

specified by the OGTR the Department of Agriculture and Water Resources and AS/NZS 2243.3:2010 guidelines are

followed.

3.3.2 Gene Technology Review Committee

According to OGTR legislation, all work involving GMOs must be reviewed by an institutional biosafety committee. The GTRC performs the function of the institutional biosafety committee. The GTRC is responsible for:

reviewing research and teaching applications which involve the use of and dealings with GMOs ensuring that the use of all GMOs within the university is conducted in compliance with the Gene

Technology Act 2000 and Gene Technology Regulations 2011

3.3.3 Heads of Units, Managers, Supervisors and Research Committees

Heads, Managers, Supervisors and Research Committees are responsible for:

ensuring that all employees and students are made aware of the biosafety manual objectives ensuring that containment facilities are suitable for teaching and research activities involving

biological hazards ensuring that staff and students are adequately trained before working in a containment facility, and

that safe work procedures are in place implementing systems to identify, assess and register biological hazards within their facility implementing a biological waste management system suitable for the containment facility they

supervise.

3.3.4 Staff and Students

Staff and students working with biological hazards must ensure:

they follow safety guidelines set out by the university, their supervisor or manager of the facility they identify any risks or hazards in their area they report unsafe conditions, difficulties, spill or containment breaches to their supervisor all hazards and incidents are reported and corrective actions implemented via the University

reporting process.

3.3.5 WHS Unit

The WHS Unit is responsible for:

providing general advice about WHS risk management facilitating provision of specialist advice and training in regards to biosafety.

3.3.6 Biosafety Representatives

The following functions and roles are carried out by the Biosafety Representatives:

attend biosafety representative meetings implement biosafety practices and resources in individual units on an ongoing basis identify risk groups in individual areas identify training needs identify system requirements

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discuss biosafety issues that may be raised in individual units with biosafety representative group assist in assessing safe work practices related to biosafety (where suitably qualified) consult with individual units on outcomes of biosafety representative meetings assist in non-OGTR PC2 laboratory inspections.

4 Definitions Biohazard: Any organism (and/or its toxin) or a material of biological origin that can cause harm to human, plants, animals and the environment. Human and Animal Diagnostic Specimen: Any human, animal, plant or invertebrate material, including, but not limited to, excreta, secreta, blood and its components, tissue and tissue fluids submitted for purposes of diagnosis or analysis. Such specimens would normally be regarded as Risk Group 2 and shall be handled in Physical Containment Level 2 facilities unless a higher risk group is indicated by the clinical notes. This applies in all microbiology and other pathology laboratories, e.g. for haematology and biochemistry. However, if a microorganism of a higher risk group is isolated from a specimen, the isolate and all samples from that source shall be handled according to the corresponding risk group, and at the appropriate physical containment level. All clinical and diagnostic specimens shall be treated with care as they may contain multiple types of infectious microorganisms. Infectious Micro-organism: A micro-organism capable of invading a susceptible host and multiplying in it, which may or may not cause a disease. Microbiological Hazard: The potential microbiological source of harm, often called a ‘biohazard’. Pathogen An infectious organism, usually microscopic, capable of causing disease in a host. Standard Precautions: The National Health and Medical Research Council (NHMRC) has recommended adoption of the term ‘Standard Precautions’ as the basic risk minimization strategy for handling of human blood (including dried blood) and body fluids, secretions and excretions (excluding sweat) regardless of whether they contain visible blood, non-intact skin and mucous membranes.(See NHMRC publication, Australian Guidelines for the Prevention and Control of Infection in Healthcare (2010)). Standard Precautions are the work practices required for the basic level of infection control. They include the use of:

good microbiological practices (e.g. aseptic non-touch techniques) good hygiene practices (particularly washing and drying hands before and after patient and

specimen contact) protective barriers (including the wearing of gloves, gowns, plastic aprons, masks, eye shields and

goggles) respiratory hygiene and cough etiquette routine environmental cleaning waterproof coverings over any break in skin integrity appropriate procedures for the handling and disposal of contaminated wastes appropriate procedures for the handling and disposing of sharps appropriate handling of linen.

The work practices described in Clause 5.3.6 of AS/NZS 2243.3:2010 (PC2 Work Practices) meet the requirements of implementing Standard Precautions.

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Diagnosis: The process of determining the nature of a disease or disorder and distinguishing it from other possible conditions.

5 The Laboratory

5.1 Containment Measures

5.1.1 General The three general descriptors by which microbiological containment is achieved are known as primary, secondary and tertiary containment measures. Optimal microbiological containment is provided by the ‘box-within-a-box’ principle, where the highest hazards are enclosed by multiple containment measures.

5.1.2 Primary containment measures Primary containment measures are the constraints immediately surrounding the source of infectious material, such as a BSC, a ventilated animal enclosure, a sealed animal room with appropriate air pressure controls, or the leak-proof container forming the inner receptacle of an approved International Air Transport Association (IATA) infectious materials transport container. Invariably, there is a primary barrier or other containment measure restricting the passage of infectious microorganisms.

5.1.3 Secondary containment measures Secondary containment measures include the design of a laboratory or device that encloses the primary containment. Facility design and engineering operations providing laboratories with air pressure control and directional air flow (supplemented by HEPA filtration of exhaust air) are examples of secondary containment measures. Another example is the secondary receptacle of an approved IATA transport container. In the laboratory or animal room, secondary physical containment measures are invariably supplemented by defined work practices, including PPE.

5.1.4 Tertiary containment measures Tertiary containment measures provide protection of the wider environment by supporting the secondary containment, e.g. using the outer packaging of an approved IATA transport container, an isolated building complex, control of people movements, and provision of support services such as decontamination and laundering of clothing and disposal of infectious wastes.

5.2 Physical Containment Classifications When working with biological hazards in a laboratory, animal house, aviary or any other room, it is necessary to define a level of containment for the facility. The Physical Containment Level of a facility dictates the equipment, work practices and Personal Protective Equipment (PPE) requirements necessary for safe work practices. There are 4 different physical containment levels ranging from Physical Containment Level 1 (PC1) to Physical Containment Level 4 (PC4). Each containment level corresponds directly to the 4 risk groups of biological hazards as set out in AS/NZS 2243.3:2010. The higher the containment level of the facility the higher the risk to laboratory workers and the community. Before working with biological hazards, it is essential to complete the Biological Hazards Risk Group Register to determine which level of physical containment is required for the facility. If it is unclear which level of physical containment is required, contact the Biosafety Representative for your area. Once the physical containment level is determined, the laboratory/room must have correct signage placed on all entrances and all appropriate procedures implemented. The risk group register must be updated annually. The Biological Risk Group register should also indicate whether any Security Sensitive Biological Agents (SSBA) are being held (see Section 5.3).

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Work involving GMOs requires prior approval from the OGTR and correct signage on laboratory entrances. The procedure to follow is outlined at http://www.uow.edu.au/research/ethics/UOW009389.html. Materials containing imported microorganisms, animals, human products, plants or soil must follow the appropriate requirements set by the Department of Agriculture and Water Resources. The Biosafety Self-Assessment Checklist is to be used by laboratory supervisors as a tool to ensure all the necessary biosafety system requirements are being implemented for each laboratory. It should be completed annually.

5.3 Laboratory Biosecurity Specific laboratory biosecurity programs should be developed for local conditions to ensure security measures are designed to prevent the loss, theft, misuse, diversion or intentional release of pathogens or toxins. Inventories of all microorganisms should be secure and personnel in such facilities should be trained in laboratory biosecurity. The National Health Security Act 2007 regulates SSBAs.

6 Administration

6.1 Contact Details Contact details for each unit’s Biosafety Representatives should be readily available to supply information and operational procedures for each laboratory.

6.2 Laboratory Access and Authorisation PC1 Facility A laboratory PC1 facility, in which laboratory personnel can be adequately protected by standard laboratory practice and no containment equipment is required, is suitable for work with microorganisms in Risk Group 1. Specimens that have been inactivated or fixed may be handled in a PC1 laboratory. PC1 laboratories must have the PC1 laboratory door sign displayed. PC2 Facility Access to any PC2 facility is restricted to authorised personnel specified by the facility supervisor. A PC2 facility, as well as displaying the PC2 laboratory sign, must display a sign identifying the access restriction to the laboratory. The facility supervisor is required to maintain a register of staff, students and others (contractors, cleaners) that are authorised to access the facility. Access to a PC2 containment facility should not be provided until the person seeking access has undertaken all the relevant training required to ensure that they can work safely in the facility. The laboratory should be kept locked at all times when not in use.

6.3 Annual PC2 Laboratory Inspections The following processes are carried out when organising and conducting an annual inspection of a non-OGTR PC2 biological laboratory (OGTR PC2 inspections are carried out annually by members from the GTRC):

1. Ideally a team of 3 members should be selected: a biosafety representative a person suitably experienced from the area being inspected a person suitably experienced from another area.

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2. Identify the laboratory representative to be present at the inspection. Inform the representative of the checklist that will be used for the inspection.

3. A suitable inspection date needs to be organised for all inspection team members and the

representative from the laboratory being inspected. 4. On the day of the inspection identify the team leader who will be given the PC2 Inspection Checklist

and conduct the inspection. The team leader will mark off questions and write comments, advise of necessary corrective actions and completion dates.

5. Once the inspection is complete a copy of the Inspection Checklist will be given/sent to the

supervisor of the laboratory. Working safely is a condition of access to the PC2 facility. Repeated failure to observe safe working practices and procedures will result in the withdrawal of access privileges.

7 Training Staff, students and visitors require training if working with biological hazards. Training includes:

7.1 Local

Induction to laboratory How to handle pathogens specific to the lab Operation of lab equipment Emergency and evacuation plans.

7.2 General Biosafety

Regulations and Standards Definitions, Risk Grouping and Facilities Work Practices PPE Equipment use Disinfectants, Transportation and Waste Immunisation and Pregnancy Biological spills management.

7.3 Genetically Modified Organisms If working in an OGTR certified laboratory, you must complete the online GMO quiz and sign off that you have read and understood the OGTR guidelines.

7.4 Training Details Please refer to the Biosafety webpage for training details.

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8 Purchasing/Acquisition Before purchasing biologically hazardous material or micro-organisms you must ensure that:

the physical containment facility is of the appropriate containment level for the risk group you are purchasing or obtaining

appropriate Department of Agriculture and Water Resources, OGTR or other relevant certificates/permits/approvals have been granted

all hazards have been identified and adequate controls are in place according to your risk assessment

you have permission from the facility manager to obtain the material. The pre-purchasing checklist can be used to assist in determining requirements before the purchase of biohazardous materials. Refer to the WHS Purchasing Guidelines for more information.

9 Risk Management

9.1 Register of Hazards The supervisor of a containment facility is required to maintain a Biological Hazards Risk Group Register. This register lists all biologically hazardous material and micro-organisms used and stored in or outside the facility, e.g. in a -80°C freezer in an adjacent area. This register is to be located in the MSDS/Hazard Register Folder or stored electronically.

9.2 Risk Assessments No research, teaching or operational work with biological hazards can be undertaken in a facility until a risk assessment of the work is conducted and it is demonstrated that all hazards are controlled. The risk assessment may be in the form of a safe work procedure for the work being undertaken.

9.3 Safe Work Procedures Safe Work Procedures should be developed for all standardised techniques, processes, equipment and machinery to minimise any risk of harm to individuals when working with biological hazards.

9.4 Incident Reporting All incidents involving biologically hazardous material shall be reported through SafetyNet in accordance with the Incident Management and Reporting Guidelines. Any laboratory acquired illness related to a biological agent stored or used in a facility must be reported via SafetyNet and reported to the faculty WHS advisor or WHS manager immediately. This includes someone developing an infected wound from a sharps injury or animal scratch or bite, or contracting a zoonotic disease from an animal handled for research or teaching purposes.

9.5 Personal Injuries All injuries are required to be reported in accordance with the Incident Management and Reporting Guidelines. Follow the procedures below in case of any of the listed events.

9.5.1 Biological Hazard Exposure to Eyes Flush the eyeball and inner eyelid with tepid water for 15 minutes. Forcibly hold the eye open to

wash thoroughly behind the eyelids Contact local first aider to get medical attention promptly.

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9.5.2 Biological Hazard Ingestion and Inhalation Contact local first aider to get medical attention promptly.

9.5.3 Biological Hazard Exposure on Skin Immediately flood the contaminated area with sufficient running water Wash area with soap and water and apply 70% ethanol or a suitable antiseptic solution. Use paper

towels to scrub affected area Remove all contaminated clothing Continue to rinse with cold water for 15 minutes. Again, wash contaminated areas with the water and

disinfectant solution but do not apply creams or lotions.

9.6 Personal Protective Clothing and Equipment (PPCE) PC1 Facility All persons entering the laboratory must wear the following PPCE, unless lesser requirements can be justified by a risk assessment:

laboratory coat or gown (a back fastening, cuffed sleeved theatre or wrap-around laboratory gown is preferable)

enclosed footwear. Note - Safety glasses, face shields and other protective devices shall be worn when undertaking tasks to protect eyes and face from splashes and other hazards e.g. contaminated or dangerous materials, or from ultraviolet light. PC2 Facility All persons entering the laboratory must wear the following PPCE, unless lesser requirements can be justified by a risk assessment:

laboratory coat or gown (a theatre or wrap-around laboratory gown is preferable). enclosed footwear

Note - Safety glasses, face shields and other protective devices shall be worn when undertaking tasks to protect eyes and face from splashes and other hazards e.g. contaminated or dangerous materials, or from ultraviolet light. When working in a biosafety cabinet the following PPCE must be worn:

a theatre or wrap-around laboratory gown must be worn when operating a biological safety cabinet gloves must be worn when working in a biological safety cabinet and when handling human blood

and body fluids.

10 Pest Control Program All physical containment facilities are required to have a pest control program against insects, birds and animals. If pest activity is identified in the laboratory, inform the laboratory manager/supervisor and request a visit by the pest control service. NOTE: Live animal or invertebrate facilities require special arrangements to prevent pests entering and/or escaping from the facility. This includes pest control in feed and bedding storage areas. See AS/NZS 2243.3:2010, Section 6 Animal Containment Facilities and Section 8 Invertebrate Containment Facilities.

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11 Facility Work Practices

11.1 PC1 Facility PC1 work practices are additional to general laboratory work practices. The following practices are to be observed when working in a PC1 facility:

no food or drink is to be consumed in the laboratory or stored in laboratory refrigerators. The application of cosmetics and shaving is prohibited in the laboratory

long hair shall be tied back all cultures must be clearly identified and dated do not store cultures for long periods on the bench. Transfer cultures to a dedicated storage area,

such as a refrigerator or part of a cold room take care to prevent the dissemination of material while flaming a wire loop, by drawing the loop from

the cooler to the hotter parts of the Bunsen burner flame, or by using a hooded or an ‘electric’ Bunsen burner

petri dish cultures of fungi must be sealed with laboratory stretch film to prevent dispersal of spores, which may be allergenic or contaminate other cultures

handle diagnostic kits and control sera with care as the exclusion of all pathogens cannot be guaranteed

take care to minimise the production of aerosols where work is carried out on the open bench take precautions to ensure that reading and writing materials do not become contaminated use self-adhesive labels clean up all spills immediately and decontaminate the area report significant spills and accidents immediately to the facility supervisor. Complete online incident

report form decontaminate work benches at least daily and after each task is completed remove laboratory gowns and store in the facility provided thoroughly wash hands and fingernails before leaving the facility.

NOTE: Hands, pens and pencils should be kept away from the face to protect from contamination due to dirty surfaces, liquids and aerosols.

11.2 PC2 Facility Work Practices The following work practices must be followed in addition to the general laboratory and PC1 work practices:

laboratory windows and doors must be closed when work is in progress used sharps, syringes and needles must be placed in the approved yellow sharps bins provided.

Before placing materials into a sharps bin, needles must not be removed, bent, sheared, or recapped

all visitors must receive an induction before they can work in the facility potentially contaminated surfaces must be disinfected before maintenance of equipment is

conducted all clinical specimens shall be regarded as potentially hazardous. Leaking containers must be

handled in a biological safety cabinet and the outside of the container disinfected. Where a replacement sample is obtainable, the leaking specimen shall be sterilised and discarded

for manipulations that produce aerosols, such as shaking, mixing, and ultrasonic disruption, a biological safety cabinet or other equipment designed to contain the aerosol must be used

a period of at least 5 min shall be allowed for aerosols to settle before opening homogeniser or sonicator containers in a biological safety cabinet

any container of viable micro-organisms transported outside the facility must be within a second unbreakable and closed container (secondary containment) which can be readily decontaminated. There should also be sufficient absorbent material (such as tissue paper) placed around the primary container to absorb any potential spill

potentially contaminated, reusable glassware must be pressure steam sterilised or chemically disinfected prior to washing and re-use. For chemical disinfection, pipettes shall be placed in a disinfectant solution, tip-first and fully immersed, to minimise the production of aerosols

special care must be taken in handling human blood, serum, other body fluids and substances that are visibly contaminated with blood, as these may contain viruses, such as hepatitis viruses or HIV.

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This risk extends to human sera and derivatives used as control reagents (both positive and negative) in diagnostic and other procedures

minor cuts, abrasion and dermatitis should be adequately covered and kept dry reference documents and papers other than worksheets must not be used on the bench bacterial cultures must not be sniffed for odours.

11.3 GMO Physical Containment Facilities Any person working with GMOs in a laboratory/room is required to follow the guidelines for containment facilities as set out by the OGTR in addition to all other requirements listed in the AS/NZS 2243.3:2010.

11.4 Quarantine Approved Premises (QAP) Any person wishing to import microorganisms, animals, human products, plants or soil for their research is required to have a Permit to Import Quarantine Material from the Department of Agriculture and Water Resources. the Department of Agriculture and Water Resources will assess if the products can be released on arrival or if they need to be used in a QAP facility. If there is a need to work in a QAP facility, conditions set by the Department of Agriculture and Water Resources must be met in addition to all other requirements listed in the AS/NZS 2243.3:2010.

12 Biological Safety Cabinets The main causes of laboratory-acquired infections are due to aerosols being produced from common laboratory operations. Biological Safety Cabinets (BSCs) provide the primary source of protection to laboratory workers against the risk of exposure to aerosols. BSCs are divided into three classes. Class I and II cabinets provide a degree of protection against work that produces significant quantities of aerosols involving microorganisms which fall into Risk Groups 2 and 3. Class II cabinets also provide a degree of product protection. Class III cabinets are used for containment of microorganisms falling into the higher Risk Groups 3 and 4 and provide a high degree of protection to the individual and product. It should be noted that laminar flow clean benches (clean workstations) only provide HEPA filtered air to protect the work being carried out on the bench. These work stations should not be used for any work involving micro-organisms or hazardous materials. It is not recommended that Bunsen burners be used in Class II cabinets as they disrupt the laminar flow and the barrier air. It is preferable to use disposable implements or electrical heating. Training in the correct use and work practices of the BSC must be completed before conducting any work in the unit. Safe work procedures for the work carried out in the BSC must be developed and followed. The following procedures should be carried out when working in a BSC:

wear a continuous-fronted gown with elasticized wrist cuffs wear protective gloves before placing items in the BSC, make sure the work area is clean - wipe down with an appropriate

disinfectant solution turn on the BSC 5 minutes before use to establish stable airflow and to purge the work zone of

aerosols ensure UV light is off prior to working within BSC arrange all required equipment and materials in the BSC within or in close proximity to the cabinet.

Where appropriate, decontaminate external surfaces of work materials before placing them in the BSC. Place work items towards the centre of the work floor so the work access opening will not interfere with the barrier air

allow the BSC to operate for another 5 minutes before commencing work follow good biological practices and sterile technique when handling materials in the cabinet slow arm movements should be used to help maintain the integrity of the air curtain in the BSC when work is completed transfer culture/s to suitable container/s for incubation or storage wipe equipment that has been used in the BSC with a suitable disinfectant solution

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wipe down the work zone with a suitable disinfectant solution remove gloves and discard in infectious waste bin. Wash hands appropriately leave the BSC operating for another 5 minutes to purge air spaces before switching off switch the BSC off, place front cover back on cabinet.

The sump of Class II BSCs should be cleaned regularly (e.g. weekly) or following a known spill. While the BSC is operating, the work floor should be lifted and the under-surface, rear grille, air intake grille, sump floor and all other accessible surfaces cleaned with suitable disinfectant solution. Refer to section 9.3 of the AS/NZS 2243.3:2010 on spills inside BSCs. BSCs must be inspected, decontaminated and tested by a NATA certified technician annually, or if the BSC is suspected of not operating correctly, or is found to have a fault. An audio-visual presentation outlining the use of BSCs can be accessed on the Victorian Infectious Diseases Reference Laboratory web site, www.vidrl.org.au

13 Working after Hours Refer to Working Alone and After Hours Work Guidelines. Follow your unit’s working after hours requirements.

14 Health Management

14.1 General All personnel shall be advised of the risk of occupational exposure to microorganisms to which they may not be immune.

14.2 Immunisation It is recommended that persons working with potentially infectious agents be immunised with the relevant vaccination if available. Refer to Immunisation Guidelines.

14.3 At Risk Persons Persons who are immuno-suppressed, immuno-compromised, or otherwise unduly vulnerable to infection, such as persons who are diabetic, should inform their supervisor so that appropriate action may be taken. Medical opinion may be required if working with human pathogens. Some microorganisms that are regarded as part of the normal flora of humans or animals may be pathogenic for immuno-compromised persons.

14.4 Precautions for Pregnant Women Pregnant women should be informed of the potential risk to the unborn child when exposed to certain microorganisms. Refer to Pregnancy at Work Guidelines.

14.5 Health monitoring All persons who will be working with laboratory animals must complete the pre-employment medical questionnaire and those that are identified as at risk must undergo an initial health assessment and regularly monitor for symptoms. Refer to the Air and Health Monitoring Guidelines. When working with pathogens of risk group 3 or 4, workers shall have an initial health assessment which includes a base line serum sample and periodic monitoring examinations dependant on the risk and level of exposure. Refer to the Air and Health Monitoring Guidelines.

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15 Personal Hygiene To prevent the spread of laboratory contaminants, it is important that hands are washed after completing a procedure, and before leaving the laboratory. See the Good Hand Washing Technique diagram (Appendix 3) for details on the correct hand washing procedure. Recommended antiseptics for hand washing can be found in Appendix 2.

16 Disinfectants For a summary of recommended chemical disinfectants for all laboratory procedures see Appendix 2.

17 Laundering Of Laboratory Gowns Laboratory gowns should be laundered on a regular basis. Before being sent to the laundry, coats used in PC2 facilities must be autoclaved or chemically disinfected. Refer to individual Units for laundry procedures.

17.1 Safe Removal of Contaminated Gloves See Appendix 4 for diagram showing how to remove gloves safely.

18 Biological Waste All laboratory waste contaminated with or potentially contaminated with microorganisms must be decontaminated, preferably by autoclaving, before it leaves UOW. Laboratory wastes are to be collected in segregated containers, clearly identified according to the following categories:

Domestic Waste - non-infectious materials, such as waste paper and plastics, that are not contaminated with microorganisms or other biohazardous materials can be disposed of in a bin labelled domestic waste (the bin contains a single layer plastic bag). This waste may be disposed of in the same manner as household waste.

Sharps – these are to be collected in an approved yellow sharps container, and can include syringes with needles, broken glass, scalpel blades and glass pasteur pipettes. Once containers are 3/4 full, they should be sealed, labelled and taken to the appropriate waste collection receptacle/store for collection.

Biological Waste – any material potentially contaminated with microorganisms, including human tissues, blood, body fluids and animal carcasses are to be placed in a waste bin labelled as biological waste. The biological waste bin contains an autoclave bag displaying the biohazard symbol, which can be pressure steam sterilized. Once waste is autoclaved, it can be disposed of in normal waste ‘sulo’ bins. See Animal Facility Manager for detailed instructions on animal carcass disposal.

GMO Waste – GMOs must be decontaminated according to the instructions set out in the OGTR requirements for a physical containment level 2 (PC2) laboratory facility. Transport of waste must be carried out in accordance with the “Guidelines for the Transport of GMOs”.

Co-Mingled Material - if any co-mingled waste is generated, contact the facility manager and the WHS Unit to assist with organising storage and disposal. Co-mingled waste includes any other hazardous waste, e.g. chemical or radiation waste that is also contaminated with microorganisms. It should be disposed of in a manner that addresses both hazards. Try to avoid generating mixed waste if possible.

Radioactive Contaminated Material – collect solid waste into robust plastic containers, labelled with isotope and date, within a secondary solid container. Collect liquid waste into container for decontamination.

Refer to the Waste Disposal Guidelines for further instructions on hazardous waste management and disposal.

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19 Emergency Management for Microbiological Spills

19.1 General If it is possible for a material to be spilled, it is likely that it will happen at some time. To control the hazard from spilled biological material, every laboratory working with biologically hazardous material (especially infectious biological agents) must develop written emergency spill/clean-up response procedures appropriate to the hazards of that material. Spills are categorised as follows:

spills inside BCSs spills in PC2 facilities outside BSCs that can be cleaned up by workers spills in PC2 facilities outside BSCs with clean up performed by trained spills clean-up team.

19.2 Factors Influencing Response to Spills The nature of a spill will determine the type of response required. Some factors include:

the amount of aerosols that have been generated from the spill whether the spill is large or small the risk group classification of the organism that has been spilled how infectious or easily transmissible the spilled organism is whether the spill is confined within a piece of equipment (such as a BSC, incubator, refrigerator,

shaker bath or centrifuge) or unconfined (on the work bench, floor of the laboratory or incubator room)

the type of surface where the spill occurred (e.g. absorbent or non-absorbent surface) the area where the spill occurred (e.g. in a contained area such as a microbiology laboratory, or in a

public access area such as a corridor or lift). The spill may also involve other hazards not of a biological nature which include:

isotopes chemicals plant/equipment electrical equipment sharps (from broken glass or equipment) liquid nitrogen/low temperature.

Identification and assessment of all the risks associated with the spill, both potential and actual, as well as the various factors listed above, must be taken into consideration before any spill clean-up begins.

19.3 Planning for the Management of Spills Planning for the spill or accidental release of biological material within the laboratory involves:

assessing and understanding the risks associated with the biological material being used (risk assessment)

ongoing training programs for staff in the correct response to accidents providing emergency response procedures and protocols for spills management providing suitable equipment for clean-up and disinfection and having available those sources of information which will help a trained clean-up group to select the

correct approach for the particular circumstances. Emergency/clean-up materials and equipment should be kept at an appropriate location outside the laboratory and should contain items appropriate to the biological material used in the laboratory. All new staff should be trained in the unit’s emergency response procedures and protocols for spill management.

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19.4 Basic Biological Spill Clean-up Kit This is an example of a basic spill kit (based on AS/NZS 2243.3:2010)

written spill clean-up procedures a "Do Not Enter" door sign with universal 'Bio-hazard' symbol protective equipment, including latex gloves, protective clothing, safety glasses, boots and

respiratory protection tape or marking device to mark off the spill area suitable disinfectant supplies - check expiry date and dilution (see Appendix 2) absorbent material (cotton balls, incontinent pads or paper towels) sharps collector and forceps for picking-up broken glass or sharps (dustpan & broom) appropriate containers or autoclave bags (disposal bags - leak proof, autoclavable and labelled with

a biohazard symbol) spare clothing for contaminated personnel.

Note: Spill kits have to be tailored to suite the type of biological material and risk group of the microorganism being used in your work area.

19.5 Clean-up Procedures Spill clean-up procedures are well documented in AS/NZS 2243.3:2010 Clause 9.3 and 9.4. Because of the importance of spill clean-up procedures, the above clauses have been set out in full.

19.6 Spills Inside Biological Safety Cabinets

19.6.1 General This procedure applies to spills that occur inside all types of biological safety cabinets, irrespective of the level or type of containment facility. Spills inside a biological safety cabinet are generally considered to be a lower hazard that those outside the cabinet as they are contained and aerosols are swept away by the cabinet air stream. Clean-up may be commenced immediately and may be done by the workers themselves.

19.6.2 Small Spills Small spills i.e. droplet-size spills or those up to 1 mL, may be treated easily by wiping with disinfectant-soaked absorbent material or flooding with a suitable disinfectant solution. Allow adequate time for the disinfectant to take effect.

19.6.3 Larger Spills The suggested procedure for a larger spill or breakage is as follows:

a) Ensure that the cabinet remains operating to retain aerosols during Steps (b), (c), (d) and (e). b) Place absorbent material wetted with suitable disinfectant or proprietary absorbent materials which

release hypochlorite over the spill. Allow approximately 10 min to effect disinfection. NOTES: 2. The three parameters that affect the efficacy of the disinfectant are concentration, time and

temperature 3. Appendix 2 lists suitable disinfectants

c) Disinfect gloved hands and remove protective gloves in the cabinet. Remove and contaminated clothing for decontamination and wash hands and arms. Replace with clean gloves and protective clothing for carrying out the remainder of the clean-up.

d) After initial disinfection of the spill, remove any sharp objects with forceps and discard as contaminated sharps then remove excess fluid with absorbent material and discard into a container for decontamination. Discard culture bottles, petri dishes and solid material associated with the spill into the same container. Decontaminate cultures, media and disposable materials adjacent to the spill.

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e) Wipe down the work floor, cabinet work zone and remaining items of equipment with fresh disinfectant solution. For Class II cabinets, disinfect both sides of the front grille and work floor within the cabinet. Check that the spillage has not contaminated the sump. If the sump is contaminated, add sufficient disinfectant solution to completely cover the sump floor. If the spill is large, use sufficient disinfectant solution to dilute and inactivate the infectious material.

f) Consider whether the cabinet should be decontaminated before further use. g) Complete an incident report in SafetyNet.

19.7 Spills Outside Biological Safety Cabinets Spills outside biological safety cabinets may be of varying degrees of complexity, ranging from spills where a limited number of persons work to those occurring in high access areas such as corridors. All efforts should be directed towards minimising the chance of a spill occurring. For PC2 and higher levels of containment, material containing microorganisms that is being moved in or between facilities or service areas shall be contained in secondary sealed, unbreakable containers. Spills can involve amounts of material ranging from 1 mL or less, to more than 100 mL. The amount spilled, the physical characteristics of the material and how the spill occurred are important factors in determining the area of involvement. When liquid is spilled, it is generally dispersed as three spill fractions:

(a) the bulk of the liquid that remains in an irregular puddle (b) the portion that separates as splashes and rivulets (c) the small portion that is separated as airborne particles.

The larger airborne particles settle rapidly, whereas the smaller particles can remain suspended in air for a considerable time and can be transported from the spill site by a ventilation system. In the event of a spill of liquid, it shall be assumed that an aerosol has been generated. All staff members should learn the basic procedure for the control of microbiological spills. Decontamination procedures for spills of infectious material shall contain the contamination in the affected area. Spills in confined areas, especially cold rooms, require special considerations e.g. the air-conditioning and air flow direction. General spills, such as from liquid cultures or culture plates, shall be treated with a suitable disinfectant. The treatment of microbiological spills in all levels and types of containment facilities shall be determined by the risk assessment. After a spill has been cleaned up, an incident report shall be completed in SafetyNet.

19.7.1 Spills in PC2 facilities that can be cleaned up by the worker AS/NZS 2243.3:2010 advises (Clause 9.4.2) that: In spills external to BSCs, low hazard (as determined by the risk assessment) infectious material that is spilled without generating significant aerosol, and does not contain a human pathogen spread by the respiratory route, should be cleaned up with a paper towel or other absorbent material soaked with an effective chemical disinfectant. The response should be as follows:

a) Remove the laboratory gown and any other garment suspected of being contaminated, and place in a biohazard bag for subsequent decontamination. If it is suspected that shoes are contaminated, remove and place in a separate biohazard bag

b) Put on appropriate protective clothing such as gloves and gowns and eye protection c) Place absorbent material wetted with suitable disinfectant over the spill. Alternatively, proprietary

absorbent materials which release hypochlorite may be used. Allow at least 10 min to effect disinfection. Remove any sharp objects with forceps and discard as contaminated sharps

d) Use the same disinfectant solution to wipe over the area likely to have been contaminated, allowing 10 min for disinfection time

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e) Carefully mop up the spill and disinfection solution, and transfer all contaminated materials for decontamination by pressure steam sterilisation

f) Remove protective clothing and decontaminate hands g) Complete an incident report in SafetyNet.

19.7.2 Spills in PC2 facilities that should be cleaned up by a dedicated spills clean-up team AS/NZS 2243.3:2010 (Clause 9.4.3) advises that: A spill external to a BSC of a large volume of high risk (as determined by the risk assessment) infectious material with the generation of aerosols will require evacuation of the area and clean-up by a trained spills clean-up team. The team shall wear protective clothing and RPE if the spill is hazardous to humans by the respiratory route. Once a spill of this type has occurred the area shall be evacuated immediately and sufficient time allowed (generally 30 min) for aerosol particles to be dispersed before contaminated surfaces are disinfected. NOTE: Although in certain circumstances respirators with P2 filters can provide adequate respiratory protection, the higher protection offered by HEPA filters with full face respirator is recommended for spill clean-up operations. Goggles should be worn where full face respirators are not used. The response for the worker should be as follows:

a) If safe to do so, contain the source of the spill. Move away from the spill b) Remove the laboratory gown and any other garment suspected of being contaminated, and place in

a biohazard bag for subsequent decontamination. If it is suspected that shoes are contaminated, remove and place in a separate biohazard bag

c) Warn others to keep out of the area of the spill d) If contamination of the worker is superficial, wash exposed skin and put on a clean laboratory gown.

Use an eye wash station if the eyes or face have been exposed e) Leave the area and place a biohazard sign with ‘DO NOT ENTER’ on the door f) Notify the area supervisor or biosafety representative of the spill g) If spilled material has soaked through the clothing, take a complete body shower in a regular i.e. not

an emergency, shower wherever possible.

The response for the spills clean-up team should be as follows:

i. Stay out of the spill area for at least 30 minutes NOTE: Consideration should be given to isolation of the recirculating ventilation systems.

ii. Assemble a clean-up team consisting of three people: one to observe and direct the clean-up procedure, and the other two to carry out the procedure. Check all necessary equipment is available

iii. Before entering the area of the spill, put on appropriate protective clothing and equipment, such as gowns, gloves, boots, eye and respiratory protection. (See Clause 10.2 of AS/NZS 2243.3:2010)

iv. Determine the extent of contamination v. Place absorbent material, such as paper towels, wetted with disinfectant, over the spill. Allow at least

10 min to effect disinfection NOTE: Appendix 2 provides a list of disinfectants

vi. Carefully remove any sharp objects with forceps and dispose of as contaminated sharps then clean up the spill and disinfectant solution. Starting from the outside, wipe towards the centre of the spill. Transfer all contaminated materials for disposal

NOTE: Waste that includes chlorine-containing materials should not be decontaminated by pressure steam sterilisation due to the likelihood of toxic fumes being produced

vii. Use the same disinfectant solution to wipe over surrounding areas likely to have been contaminated with aerosols. Allow 10 min for disinfection time then discard waste for decontamination

viii. Ensure that each member of the clean-up team decontaminates PPE used to clean up the spill ix. Complete an incident report in SafetyNet.

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19.8 Centrifuge Spills AS/NZS 2243.3: 2010 (Clause 9.5) states that: Where a spill or leak is detected within a centrifuge, the procedure will depend upon the risk group of the agent involved (see Clause 9.1) as well as the construction of the equipment. The clean-up procedure should be as follows:

a) Sealed rotors or buckets that can withstand high temperatures - Thermally decontaminate intact at 121°C for a minimum of 15 min. (See Appendix 2)

b) Rotors and buckets not able to withstand high temperatures – Where breakage or spillage is observed, allow 30 min for aerosols to settle. Place the rotor or bucket in an appropriate non-corrosive disinfectant solution (See Appendix 2). If the disinfectant is corrosive, wipe internal surfaces with water or detergent at the end of the contact time. The use of glass centrifuge tubes should be avoided. If a glass centrifuge tube has broken, remove larger pieces of broken glass to the sharps container with forceps and use material such as cotton wool moistened with disinfectant to pick up the finger pieces. Wipe internal surfaces of the centrifuge bowl with disinfectant.

19.9 Disposal of Contaminated Waste Following Spill Clean-up Normal laboratory waste disposal procedures should be used for spill clean-up material. Spill clean-up material containing bleach should not be autoclaved. Disinfectants should not normally be autoclaved as they can damage the autoclave or produce toxic vapours.

20 Fieldwork Biological hazards from some flora and fauna may be encountered when working in the field. Bites from various insects and animals may cause poisoning and disease. Some plants have the capacity to sting and cause infections. Other biological hazards associated with fieldwork include:

sanitation requirements hygiene Immunizations.

Refer to Fieldwork Guidelines and the Scuba Diving Operations Manual for further information.

21 References

Australian/New Zealand Standard 2243.3:2010 Part 3: Microbiological safety and containment Australian/New Zealand Standard 2252.1:2002 Biological safety cabinets - Biological safety cabinets

(Class I) for personnel and environment protection Australian/New Zealand Standard 2252.2:2009 Controlled environments – Biological safety cabinets

Class II - Design Australian/New Zealand Standard 2252.4:2010 Controlled environments – Biological safety cabinets

Classes I and II – Installation and use (BS 5726:2005, MOD) Australian Immunisation Procedures Handbook (10th Edition, NHMRC) Guidelines for Certification of PC2 Facilities/Physical Containment 2 requirements Version 3.2 – 1st

March 2013 Australian Guidelines for the Prevention and Control of Infection in Healthcare (2010) (National

Health and Medical Research Council) National Code of Practice for the Control of Work-related Exposure to Hepatitis and HIV (blood-

borne) Viruses [NOHSC:2010 (2003) 2nd Edition] Quarantine Approved Premises for Imported Biologicals (Department of Agriculture and Water

Resources)) WorkCover NSW – Pregnancy and Work – Guide 2002 Laboratory Safety Guidelines Biological Hazards Risk Group Register Biological Self-Assessment Checklist PC2 Inspection Checklist WHS Purchasing Guidelines Pre-Purchase Checklist

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Incident Management and Reporting Guidelines Working Alone and After Hours Work Guidelines Immunisation Guidelines Pregnancy at Work Guidelines Waste Disposal Guidelines Fieldwork Guidelines Scuba Diving Operations Manual Air and Health Monitoring Guidelines

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22 Version Control Table Version Control Date Released Approved By Amendment 1 May 2006 WHS Manager New manual created 2 Oct 2006 WHS Manager Manual updated 3 Dec 2006 WHS Manager Manual updated 4 Aug 2008 WHS Manager Manual updated 5 August 2010 WHS Manager Document updated to incorporate the

Personnel name change to Human Resources Division.

6 March 2011 WHS Manager Document updated to reflect changes to the Australian Standards referred to throughout the document.

7 March 2012 WHS Manager Rebrand 8 August 2012 WHS Manager Training requirements updated to reflect the

move to internal training courses. 9 May 2013 WHS Manager Document updated to reflect unit name

change to WHS and to update the change in legislation. Requirements for displaying PC1 laboratory door signs added.

10 October 2015 WHS Manager Document updated added health monitoring requirements and changed AQIS section to Department of agriculture and water resources

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Appendix 1 – Biosafety Legislation and Guidance Legislation (Commonwealth) Biological Control Act 1984 Gene Technology Act 2000 Gene Technology Regulation 2011 Prohibition of Human Cloning Act 2002 Biosecurity Act 2015 Research Involving Human Embryos Act 2002 Crimes (Biological Weapons) Act 1976 Weapons of Mass Destruction (Prevention of Proliferation) Act 1995 Crimes (Biological Weapons) Regulation 1980 National Health Security Act 2007 National Health Security Regulations 2008 Legislation (NSW) Gene Technology (NSW) Act 2003 NSW Work Health and Safety Act 2011 NSW Work Health and Safety Regulation 2011 NSW Occupational Health and Safety Amendment (Dangerous Goods) Regulation 2005 Biological Control Act 1985 Anatomy Act 1977 Human Tissue Act 1983 Human Cloning for Reproduction and Other Prohibited Practices Act 2003 Research Involving Human Embryos (NSW) Act 2003 Animal Research Act 1985 Protection of the Environment Operations Act 1997 Public Health Act 1991 Public Health (Microbial Control) Regulation 2000 Animal Research Regulation 2010 Public Health (Disposal of Bodies) Regulation 2002 Guidelines for the Funeral Industry (Based on the Public Health (Disposal of Bodies) Regulation 2002 Australian Standards Australian/New Zealand Standard 2243.3:2010 Part 3: Microbiological safety and containment Australian/New Zealand Standard 2982:2010 Laboratory design and construction Australian/New Zealand Standard 2252.1:2002 Biological safety cabinets - Biological safety cabinets (Class I) for personnel and environment protection Australian/New Zealand Standard 2252.2:2009 Controlled environments – Biological safety cabinets Class II - Design Australian/New Zealand Standard 2252.4:2010 Controlled environments – Biological safety cabinets Classes I and II – Installation and use (BS 5726:2005, MOD) Australian/New Zealand Standard 3666.2:2011 Air-handling and water systems of buildings - Microbial control - Operation and maintenance Australian/New Zealand Standard 3666.3:2011 Air-handling and water systems of buildings - Microbial control - Performance-based maintenance of cooling water systems Australian/New Zealand Standard 3816:1998 Management of clinical and related wastes Codes of Practice National Code of Practice for the Control of Work-related Exposure to Hepatitis and HIV (blood-borne) Viruses [NOHSC:2010 (2003) 2nd Edition] Australian code of practice for the care and use of animals for scientific purposes 7th edition 2004

Other WorkCover NSW – Dermatitis (The Facts Starting From Scratch) WorkCover NSW - Guidelines for Assessing the Risk of Exposure to Biological Contaminants in the Workplace WorkCover NSW – Pregnancy and Work – Guide 2002 Human Research Ethics Handbook Environmental Guidelines: Assessment, Classification & Management of Liquid & Non-Liquid Wastes (Department of Environment and Climate Change, NSW) Australian Immunisation Procedures Handbook (10th Edition, NHMRC)

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Australian Guidelines for the Prevention and Control of Infection in Healthcare (2010) (National Health and Medical Research Council) Handbook on the Regulation of Gene Technology in Australia (Office of the Gene Technology Regulator) Guidance Note on the Interpretation of Exposure Standards for Atmospheric Contaminants in the Occupational Environment [NOHSC:3008(1995)] 3rd Edition Guidance Notes for the Transport of Class 6.2 (Infectious Substances) Dangerous Goods (Department of Infrastructure and Transport) Quarantine Approved Premises for Imported Biologicals (Australian Quarantine and Inspection Service) International Air Transport Association

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Appendix 2 - Summary of Recommended Applications for Chemical Disinfectants in Microbiological Laboratories Refer to AS/NZS 2243.3:2010 Appendix F for further information.

Site or Equipment Routine or Preferred Method or Usage

Acceptable Alternative

Benches and surfaces (not obviously contaminated)

Alcohols e.g. 70% w/w (= 80% v/v) ethyl or 60–70% v/v isopropyl – swabbed (See paragraph F6.9)

Synthetic phenolics (See paragraph F6.10)

Biological safety cabinet (BSC) work surfaces

Alcohols e.g. 70% w/w (= 80% v/v) ethyl - swabbed or high concentration chlorine disinfectant at 5000-10 000 p.p.m. (0.5 − 1%) (See paragraph F6.1) or other disinfectant depending on the organism

For BSC with capture hoods, glutaraldehyde† (with cabinet fan operating)—swabbed (see AS/NZS 2647)(See paragraph F 6.4)

Room space e.g. laboratory or animal room, BSC before servicing or testing or after major spill

Formaldehyde vapour (see Paragraph F6.3)

Vaporised hydrogen peroxide (see paragraph F6.7) or Chlorine dioxide (see paragraph F6.8)

Centrifuge rotor or sealable bucket after leakage or breakage

Disinfection not the preferred method. Pressure steam sterilizing at 121°C for 15 min recommended (See Clause 10.6)

Glutaraldehyde† for 10 min or synthetic phenolics* for bacterial spills for 10 min(See paragraph F 6.10)

Centrifuge bowl after leakage or breakage

Glutaraldehyde† for 10 min (swabbed twice within the 10 min period then wiped with water) )(See paragraph F 6.4)

Synthetic phenolics* for bacterial spills for 10 min )(See paragraph F 6.10)

Discard containers (pipette jars) Chlorine disinfectant at 2 000–2 500 p.p.m. (0.2–0.25%),

Synthetic phenolics* for bacteriological work (changed weekly) or detergent with pressure steam sterilizing for virus work

Equipment surfaces before services or testing

Surfaces disinfected according to manufacturers’ instructions

Alcohol (80% v/v ethyl or 60–70% v/v isopropyl) except when its flammability poses a hazard (See F 6.9)or glutaraldehyde† then water (see F6.4)

Gnotobiotic animal Isolators Peracetic acid at 2% v/v —swabbed (see F6.5)

Hand disinfection Chlorhexidine (0.5–4% w/v) in alcoholic formulations for 2 min(See F6.12)

Isopropyl (60–70% v/v) or ethyl alcohol (80% v/v) ) (See F 6.9) with emollients or Povidone-iodine (0.75–1% av I) for 2 min(See F6.2)

Hygienic hand wash Chlorhexidine (4% w/v) in detergent formulation (or alcoholic formulations) for 15 s (See F6.12)

Detergent cleansers or soap for 15 s

Spills of blood/serum (or viral cultures)

High concentration chlorine at 5000– 10 000 p.p.m. (0.5–1%) (See F6.1) for 10 min (active against hepatitis viruses and HIV)

Glutaraldehyde† for 10 min (See F6.4)

Spills of bacterial cultures High concentration chlorine disinfectant at 5 000–10 000 p.p.m. (0.5–1%) or Iodophor* for 10 min (See F6.2)

Synthetic phenolics*(unaffected by organic load) for 10 min (See F6.10)

Spills of bacterial cultures High concentration chlorine disinfectant at 5000-10 000 p.p.m. (0.5 − 1%)* (See F6.1) or Iodophor* for 10 min (See F6.2)

Synthetic phenolics* (unaffected by organic load) for 10 min (See F6.10)

Animal cages Wash with detergent followed by pressure steam sterilizing at 121°C for 15 min if infected (See Clause 10.6)

Drains and animal rooms (surfaces) Sodium hydroxide 1M

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Appendix 3 - Good Hand Washing Technique

Step 1 : Thoroughly wet hands with water

Step 2 : Apply soap or antiseptic Step 3 : lather by rubbing hands together vigorously

Step 4 : Palm over back of hand

Step 5 : Palm to palm with interlaced fingers

Step 6: tips and backs of fingers to each palm

Step 7 : Scrub fingernails on palm Step 8 : Clean Thumbs

Step 9 : Clean wrists Step 10 : Rinse hands thoroughly Step 11 : Pat dry hands thoroughly Note: the whole hand washing process should take around 15 seconds.

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Appendix 4 - Removing Gloves Safely

Step 1 : Ensure you are near a bio-waste bin before taking off gloves

Step 2 : Pinch glove on palm Step 3 : slowly pull glove down Step 4 : Pull glove completely off hand

Step 5 : do not touch glove with bare hand

Step 6 : Tightly scrunch glove in hand Step 7 : Put index finger under the top of the glove

Step 8 : Turn finger 180o

Step 9 : Pull glove outwards and towards fingertips

Step 10 : make sure scrunched glove remains inside glove

Step 11 : Hold glove by uncontaminated surface and dispose in appropriate waste container