i
SAFETY IN RESEARCH AND TEACHING MANUAL
MURDOCH UNIVERSITY
VERSION 2
15 July 2015
Review Date: This manual expires on XXXX
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CONTENTS
1
THE AIMS OF THIS MANUAL ........................................................... 1
2
RESPONSIBILITIES ........................................................................ 1
2.1
2.2
3
RISK ASSESSMENT FOR MURDOCH PROCESSES .............................. 1
3.1
3.2
3.3
3.4
4
MANAGEMENT’S RESPONSIBILITY.................................................................. 1
EMPLOYEES/STUDENT’S RESPONSIBILITY......................................................... 1
THE RISK ASSESSMENT PROCESS.................................................................. 2
RISK IDENTIFICATION .............................................................................. 2
ASSESSING THE LIKELIHOOD AND CONSEQUENCE OF AN ADVERSE EFFECT TO I) YOU, II)
OTHERS OR III) THE ENVIRONMENT ............................................................... 2
DEVELOPING STRATEGIES TO REDUCE THE RISK................................................. 2
OCCUPATIONAL SAFETY ................................................................. 3
4.1
WORKING ALONE AFTER HOURS .................................................................. 3
4.2
REPORTING HAZARDS AND ACCIDENTS ........................................................... 3
4.3
EMERGENCY PROCEDURES .......................................................................... 3
4.4
LOCATION OF EMERGENCY SAFETY FEATURES .................................................... 4
4.4.1
Fire extinguishers ......................................................................... 4
4.4.2
Fire blankets ................................................................................ 4
4.4.3
First aid kits ................................................................................. 4
4.4.4
Safety shower and eye wash station ............................................... 4
4.4.5
Antidotes ..................................................................................... 5
4.5
KEEP THE WORKPLACE CLEAN AND TIDY .......................................................... 5
4.6
CARE OF EQUIPMENT ITEMS ........................................................................ 5
4.7
GENERAL LAB ETIQUETTE FOR COMMUNAL LABS ................................................. 5
5
GENERAL LABORATORY SAFETY PRACTICES ................................... 7
6
WORKING WITH BIOLOGICAL MATERIAL ....................................... 9
6.1
OVERVIEW ........................................................................................... 9
6.2
INTRODUCTION ...................................................................................... 9
6.3
MICROORGANISMS .................................................................................. 9
6.4
FINDING OUT THE RISK GROUP .................................................................... 9
6.5
THE RISK ASSOCIATED WITH RESEARCH.......................................................... 9
6.6
LABORATORY PRACTICES TO BE OBSERVED WHEN WORKING WITH MICROORGANISMS ... 11
6.7
WORKING WITH GENETICALLY MODIFIED ORGANISMS (GMO’S) .......................... 11
6.8
WORK PRACTICES TO BE FOLLOWED WHEN DOING RESEARCH WITH GMO’S .............. 12
6.9
WORKING WITH LABORATORY ANIMALS ........................................................ 13
6.10 PRACTICES FOR WORKING WITH LABORATORY ANIMALS ...................................... 13
6.11 IMPORTED BIOLOGICAL OR BIOLOGICAL SOURCE MATERIAL ................................ 14
6.12 CLASSIFICATION OF LABORATORIES FOR BIOLOGICAL RESEARCH .......................... 15
6.13 HOW DO I KNOW IF MY LABORATORY IS PC1, PC2 OR PC3? ............................... 16
6.14 PREVENTING INFECTIONS ........................................................................ 16
6.14.1 Bio-safety cabinets ..................................................................... 16
6.14.1.1
6.14.1.2
6.14.1.3
6.14.1.4
6.14.1.5
Class I Bio-safety cabinet................................................................ 16
Class II Bio-safety cabinet .............................................................. 16
Class III Bio-safety cabinet ............................................................. 16
Laminar Flow Hoods ....................................................................... 17
Fume Hoods .................................................................................. 17
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6.14.2
6.14.3
Centrifugation ............................................................................ 17
Disinfectants .............................................................................. 17
6.14.3.1
6.14.3.2
6.14.3.3
Types ........................................................................................... 18
Activity of chemical disinfectants ..................................................... 20
Treatment of Laboratory Spills......................................................... 20
6.15 WASTE SEGREGATION AND DISPOSAL .......................................................... 21
6.15.1 Sharps: Syringes, Needles and Blades ........................................... 21
6.15.2 Disposal of Sharps ...................................................................... 21
6.15.3 Decontamination and Disposal of Biologicals .................................. 21
6.15.3.1
6.15.3.2
6.15.3.3
6.15.3.4
6.15.3.5
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Autoclaving ................................................................................... 21
Autoclaving solid waste .................................................................. 22
Autoclaving liquid waste ................................................................. 22
Steam treatment ........................................................................... 23
Incineration .................................................................................. 23
WORKING WITH CHEMICALS AND SUBSTANCES .......................... 24
7.1
KNOW THE HAZARDS - READ THE SAFETY DATA SHEETS (SDS) ........................... 24
7.1.1
Accessing SDS through ChemWatch .............................................. 24
7.1.2
Completing a Hazardous Substances Risk Assessment Form (HSRA) . 25
7.2
LABELLING ......................................................................................... 26
7.2.1
Decanted chemicals/solutions ...................................................... 27
7.3
PREPARED SOLUTIONS/CHEMICALS ............................................................. 27
7.4
SAFE STORAGE OF CHEMICALS ................................................................... 27
7.4.1
Handling and Storage.................................................................. 27
7.4.1.1
7.4.1.2
Precautions for safe handling .............................................................. 27
Conditions for safe storage, including any incompatibilities ..................... 28
7.4.2
Incompatible chemicals ............................................................... 28
7.4.3
Storage height ........................................................................... 28
7.4.4
Fridges/freezers and phenol or flammable material ......................... 28
7.5
CHEMICALS WITH SPECIAL HANDLING REQUIREMENTS........................................ 29
7.5.1
Carcinogens, teratogens and mutagens ......................................... 29
7.6
COMPRESSED GASSES ............................................................................ 29
7.7
DISPOSAL OF CHEMICAL AND SUBSTANCE WASTE ............................................ 29
7.7.1
Organic Solvents ........................................................................ 30
7.7.2
Inorganic Chemicals .................................................................... 30
7.7.3
Organic Chemicals ...................................................................... 30
8
WORKING WITH RADIOISOTOPE ................................................. 31
8.1
INDIVIDUAL USER RESPONSIBILITIES: ......................................................... 31
8.2
RADIOACTIVE SUBSTANCES LABORATORY GUIDELINES AND PROCEDURES ................ 31
8.2.1
Who can work in the laboratory? .................................................. 31
8.2.2
Bringing radioactive samples to the lab and storing them: ............... 31
8.2.3
Radiation monitoring: ................................................................. 32
8.2.4
Disposal of radioactive substances: ............................................... 32
8.2.5
General laboratory procedures ..................................................... 32
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FIELDWORK .................................................................................. 33
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1 THE AIMS OF THIS MANUAL
The aims of this manual are:
To make you aware of the risks associated with undertaking teaching
and research.
To provide some information on how to manage these risks.
To provide some information on the regulations associated with
working with specific materials e.g. specific chemicals or Genetically
Modified Organisms.
To indicate sources of further information that can be helpful in
assessing the risks.
2 RESPONSIBILITIES
2.1 Management’s responsibility
Ensure that laboratory workers and research students are given
training in the safety practices and safety equipment in the laboratory.
Ensure that all staff and students working in the lab have attended the
Safety in Research and Teaching Workshop.
Provide protective equipment.
2.2 Employees/student’s responsibility
To familiarise themselves with the regulations governing their research
activities and ensure that the regulations are complied with.
To familiarise themselves with the safety procedures of the laboratory,
and the safety equipment in the laboratory.
Attend relevant safety training courses offered by the Employer or
external provider.
Carry out safe work practices.
To use protective equipment to minimise injury, not only to
themselves, but also to their colleagues (and the wider community).
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RISK ASSESSMENT FOR MURDOCH PROCESSES
Before you begin your project you are required to complete a “Risk Assessment for
Murdoch Processes” form (RAMP). This form is treated as a register of the key risk
areas (chemical, biological ethical, fieldwork, etc.) for which specific approvals are
likely to be needed in the above project. For each of these there is a supplementary
risk assessment that you are required to fill in and email to the relevant approving
body. Each of the individual approvals for the project will be recorded against this file
number.
The RAMP and the supporting forms are available from the Safety in Research and
Teaching website.
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3.1
The risk assessment process
Risk assessment is a process aimed at identifying the risks associated with your
research, and of seeking ways in which the risks to the environment yourself and to
others can be minimized, or managed to reduce the risk. Risk assessment occurs in
three stages.
1.
Risk identification.
2.
Assessing the likelihood of an adverse effect to i) you or ii) to others
iii) to the environment.
3.
Developing strategies to reduce or manage the risks.
3.2 Risk Identification
Firstly don’t assume because “your supervisor said so” or “it is usual in this lab” that
a procedure is risk free. Almost no process is risk free; it is the likelihood and
severity of the risk that is most important.
The identification process involves breaking down the project into a number of
components and assessing the risks associated with each component. Obvious
examples of risks are the risks associated with using the microorganisms or
chemicals. The scale of use is also a risk; growing a 100L batch culture of an
organism is very different from growing it in a couple of Petri dishes. Even a
relatively harmless organism such as E coli can be toxic at high concentration.
Similarly for the use of chemicals, the RAMP attempts to guide you through this
process by asking a series of questions relating to whether specific components are
used in the project. The potential risks of fieldwork, in all its forms, are often
seriously underestimated, especially as the potential consequences to life and limb
can be much higher in remote locations than in the laboratory setting.
3.3 Assessing the likelihood and consequence of an adverse effect to
i) you, ii) others or iii) the environment
By this you should be asked to estimate whether there is a high, medium or low
chance of an adverse outcome of the risk happening based on consideration of both
the likelihood and consequence of a risk occurring. If you are dealing with large
numbers of people, amounts of a toxic chemical, or microorganism, then the risk of
an adverse outcome are magnified. On the other hand if you are dealing with small
numbers or amounts in a relatively benign setting or type of organism such as E coli
(found in all of our guts) the risks are quite low (provided reasonable precautions are
observed). Similarly driving to Bunbury for a couple of days is very different to
driving into the Pilbara for an extended survey trip.
3.4 Developing strategies to reduce the risk
You should identify ways in which the risks can be reduced. An obvious way is
substitution, e.g. if you are dealing with a pathogenic organism can you substitute a
non-pathogenic strain? If things do go wrong do you know what to do, e.g. if you
spill a culture of your microorganism what would you do, who would you tell, how
would you report it? For chemicals, have you read the SDS and are you aware of
how to treat accidental exposure, do you have the required reagents on hand to
treat such exposures, or to neutralize the chemical. Another strategy that reduces
the risks to you and to others is the use of appropriate equipment, biosafety hoods,
closed centrifuge bottles to prevent aerosols, personal protective equipment etc. In
the fieldwork context, are you carrying spares, a first aid kit, have you regularized
your contacts so people know if you are delayed, or injured?
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OCCUPATIONAL SAFETY
4.1 Working Alone After Hours
After hours work is any carried out before 8 a.m. and after 5 p.m. Monday-Friday
and any time on weekends and Murdoch public holidays or shut-down periods.
Several facilities at Murdoch require that anyone working after hours must sign in
when they arrive and sign out when they leave the lab. Check whether you need to
enforce this style of control on your facility.
Regardless of the above, you should always try to minimize the carrying out of
hazardous procedures after hours. Any hazardous work carried out after hours
require a minimum of 2 people working in the same general area. Hazardous
activities include the use of toxic chemicals, flammable solvents, liquid nitrogen and
radioactive material. Even heavy lifting such as connecting a boat trailer can cause
serious problems if you are on your own. This is for your own benefit - imagine how
you would cope with a fire, injury or large radioactive spill with no one else around.
In general, junior staff and students must not be allowed to work outside normal
working hours for your area. Trained persons are permitted to work outside normal
work hours provided a risk assessment of the activity has been conducted for the
hours being worked. Please refer to the Working Alone or in Isolation Policy for
further information.
4.2 Reporting hazards and accidents
If an accident occurs (however minor), report it to your supervisor, and use the
Murdoch University Incident Reporting System to report the accident. If a “near
miss” occurs, or you notice something that is potentially dangerous, notify your
supervisor or the relevant lab manager and OSH representative, and use the
Murdoch University Incident Reporting System to fill out a hazard report. This may
prevent a serious accident to yourself or other lab workers in the future.
4.3 Emergency procedures
There is no time in the middle of an emergency to start learning what you have to
do, so familiarize yourself in advance with emergency, evacuation and fire
procedures. The safety noticeboard in the facility has details of the evacuation
alarms, route and mustering point, as well as emergency contact names and
numbers. There is also a list of people in this building with first aid experience who
can be contacted in an emergency. Read the notice board from beginning to end and
also make sure you know who to contact in the event of an emergency.
Make sure you check the location of the nearest safety noticeboard to your lab.
You are required to call ‘000’ in the event of a life-threatening medical situation oncampus. For all other assistance the Murdoch University ‘333’ (internal calls)
security number is to be called, if calling from a mobiles dial 9360 7333.
If ‘000’ assistance is first sought, Murdoch Security are to be advised of the incident
by calling ‘333’ immediately following the ‘000’ call, to allow them to provide
responsive on-campus support and assist the ‘000’ response as required.
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4.4
Location of emergency safety features
4.4.1 Fire extinguishers
Ensure that you know the location of the extinguishers and also how to operate
them.
4.4.2 Fire blankets
In some instances fire blankets can be an effective alternative and more effective
solution, so familiarise yourself with their location.
Think - Where is the nearest fire blankets?
4.4.3 First aid kits
Make sure you know the location of the nearest first aid kit.
from the kit let your supervisor know.
If you use anything
Think - Where is the location of the nearest first aid kit to my laboratory?
4.4.4 Safety shower and eye wash station
If you spill a corrosive, radioactive or poisonous chemical on yourself you should get
to an emergency shower as quickly as possible (preferably a deluge shower).
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Think - Where is the nearest Shower and Eyewash Station?
4.4.5 Antidotes
Note that some poisons have antidotes that work better than water alone, for
example polyethylene glycol (PEG) should be used to aid the rinsing of phenol from
the skin. Use the SDS to work out whether you need to make up a solution in
advance that will help if you have an accident - it is way too late once a spill has
occurred.
4.5 Keep the workplace clean and tidy
Keeping the workplace clean and tidy and free of clutter and always wiping down
surfaces after using them (even if you can’t SEE any chemical residue) is one of the
most important safety steps you can take. This is especially true of communal
benches, which everyone leaves for everyone else to clean. Ensure that they are
done on a regular basis and not just when a spill occurs. Every drop of carcinogenic
liquid left to evaporate leaves yet another layer of accumulated residue on the
bench. After working with bacteria it is necessary to swab the work area with ethanol
to kill any that remain.
Note that the floors can become very slippery when wet! Whenever water is spilled
on the floor it must be cleaned up immediately. Falls can cause serious back injury,
not to mention the added danger if the person is carrying a hazardous substance at
the time.
4.6 Care of equipment items


Under no circumstances should any person use a piece of equipment that they
have not been trained to use. This is to protect the safety of the person and the
integrity of the often valuable equipment. A great deal of damage can be done to
very expensive equipment but an untrained person.
Always ask suitably qualified people to show you how to use equipment you are
unfamiliar with before you use it. Most pieces of equipment have instruction
sheets on the walls above them. Read all sheets before using the equipment,
even if someone has shown you how to use it. Report any malfunctioning or
damaged equipment to your supervisor and laboratory manager as soon as
possible.
4.7 General lab etiquette for communal labs
In communal lab areas it is essential that everyone co-operates to maintain a wellstocked, clean, tidy and safe working environment. It does not take much if
everyone pitches in, but it is a mammoth task if only one or two dedicated people
are doing it on their own. People who do not do their share cause resentment and an
eventual breakdown in the system. At a minimum, everyone should follow these
basic rules:
 If you make a mess, clean it up immediately; do not leave it for someone else.
Areas of particular concern are the balances, where your chemicals may
contaminate someone else’s experiment. The next person to come along will not
know what the contaminant was, whether it is hazardous or how to dispose of it
safely.
6
 If you notice a problem with a piece of equipment, report it to your
supervisor immediately to minimise down time, don’t wait for someone else to do
it. Leaving an “out of order” note on the equipment as well will save time for your
co-workers.

You must label every solution that you make with your full name, the date
(including the year), the contents and any hazard warning stickers required, so
that if someone accidentally knocks it over and it spills on them they know what
to do about it. If the container is too small to label (e.g. 0.2 µL Eppendorf), label
the rack instead.
Refer to SDS for particularly dangerous or volatile chemicals.
Please clear out all of your material when you leave the lab, including contents of
fridge, boxes in cold room, your bench and (especially) the -80ºC freezer. You should
check with your supervisor before discarding biologicals (organisms, plasmids
libraries, etc.) and dispose of all material in an appropriate way.
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5 GENERAL LABORATORY SAFETY PRACTICES
The following are a set of practices which should always be followed when working in
a research lab.
Table 1: General safety practices to be observed in the laboratory
Plan your work, don’t rush, and make sure you have
Planning
received instruction in the use of equipment.
Food and drink
The consumption of food and drink is strictly forbidden in
laboratories.
Food and drink must not be brought into laboratories under
any circumstances. They are to be stored in approved
eating and drinking areas ONLY. They are never to be
stored in the laboratory, or in cold rooms used for research
materials.
The application of cosmetics and the taking in or out of
contact lenses are also not permitted in the laboratory.
Hair
Long hair should be tied back as it is a hazard in the
laboratory. It can get caught up in the flame from a Bunsen
burner, get entangled in equipment, and restrict lateral
viewing.
Protective clothing
Front or back fitting lab coats should be at all times.
Where infectious materials have been handled, lab coats
MUST be autoclaved before sending to the laundry.
Lab coats must NOT be worn outside the lab and NEVER in
areas such as common rooms where food is consumed.
You should always wear covered footwear that covers the
upper part of the foot as well as the sole in the lab. Some
laboratories and certain fieldwork situations require specific
styles of footwear (steel capped work boots).
Safety glasses must be worn in laboratories where indicated
by signage.
NOTE: contact lenses are not a suitable substitute for safety
glasses. Safety glasses can be purchased from several
places inside the University, please refer to your supervisor
or lab manager for advice on where to purchase safety
glasses.
General
safety
Wash your hands before leaving the lab.
8
procedures
Mouth pipetting is strictly prohibited at all times and in all
situations.
Gloves must be worn for many procedures. Rings or hand
jewellery which would interfere with glove functioning
should be removed before putting on gloves. Gloves should
be appropriately decontaminated prior to disposal with other
laboratory wastes. Many labs have separate disposal
procedures for biological and general rubbish – always
check!
Work surfaces must be cleaned and decontaminated with a
suitable disinfectant at the end of the day and after any spill
of potentially dangerous material. PC2 or PC3 labs may
have special contingency requirements; you should contact
your supervisor or lab manager to familiarize yourself with
these, if appropriate. Loose or cracked work surfaces must
be reported to your supervisor or lab manager.
Accidents, near misses and overt or potential exposures
must be reported in writing to the laboratory supervisor or
acting alternate as soon as circumstances permit; this
person should file this report with management and the
appropriate biosafety officer or committee. Appropriate
medical evaluation, surveillance, and treatment should be
sought and provided as required. Actions taken to prevent
future occurrences should be documented.
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6 WORKING WITH BIOLOGICAL MATERIAL
6.1 Overview
The purpose of this section of the manual is to help you with rules that should apply
across the board for activities that involve working with biological material. Your
specific area of activity may require additional support documentation and standard
operating procedures to ensure that your work complies with more specific
regulations. You should discuss this with your supervisor or the SRTC if you have
any concerns. You as an individual are responsible for your safety and you should
not be involved with any activity that you have not been fully informed about and for
which you have not received appropriate training. If you supervise other staff or
students, you are also responsible for the health and safety and training of those
staff and to provide standard procedures to ensure that a safe work environment is
maintained. Some activities are regulated by government bodies and activities are
strictly controlled by law, failure to comply with these laws can have far reaching
consequences for you as an individual, the university and the environment.
6.2 Introduction
Biological material means biological material in its broadest context. Sometimes the
biological content may not be immediately apparent for example a project using
Titanium dioxide to purify waste water sounds pretty biological free but waste water
can contain a plethora or biological molecules and microorganisms that can act as
toxins, cause allergy and cause disease. The most severe biological risks come from
working with microorganisms or material that contains microorganisms.
6.3 Microorganisms
Microorganisms encompass bacteria, viruses, fungi, protozoa, mycoplasmas, and
parasites. Microorganisms covers an incredible number and diversity of species and
range from highly pathogenic forms that cause severe and debilitating diseases to
those that are quite innocuous or even beneficial.
When working with a microorganism it’s a good idea to know if it’s dangerous and
how dangerous it is. Is it likely to cause disease in humans, animals or plants or is it
a non-disease organism.
Various governmental agencies have developed a
classification scheme whereby microorganisms are categorized according to the risk
they pose. The categories range from 1 (harmless) to 4 (highly dangerous). This
should be assessed by the IBC via the RAMP process.
6.4 Finding out the risk group
If you don’t know the risk group of the organism you are working with, please refer
to the IBC website.
6.5 The risk associated with research
There are risks associated with using biological and chemical materials to undertake
research. The risks are not only to you as a researcher, but to others in the
laboratory and outside the laboratory. There is also the risk of environmental
10
damage, so that even material that seems to be innocuous may often be quite
harmful. Outlined below are some of the risks associated with working with various
materials.
Table 2: Risks associated with working with various materials
Material
Risk to Human Health
Risk
to
Environment.
Blood,
sputum,
urine,
faeces, body fluids, animal
tissue, animals (diseased
or healthy)
Risk of contracting an
infectious disease leading
to severe debilitation or
worse.
Unintended
spread
animal diseases.
the
of
Risk of transmitting an
infectious
disease
to
others (including animals)
both within and beyond
the
bounds
of
the
laboratory.
Imported animals
Risk
of
zoonoses
exposure
to
May
harbor
exotic
pathogens which could
cause epidemics in native
animal species.
Animals could become a
pest, e.g., rabbits.
Microorganisms, (cultures
of bacteria, fungi, viruses,)
or
material
containing
microorganisms
eg
sewage,
water,
soil,
compost, plant tissue.
Risk of contracting an
infectious disease leading
to severe debilitation or
worse.
Plant material.
(include
imported plants and plant
material
Suffer from exposure to
allergins and toxins/
Risk
of
transmitting
pathogenic
microorganisms to natural
populations beyond the
laboratory.
Risk of transmitting an
infectious
disease
to
others (including animals)
both within and beyond
the
bounds
of
the
laboratory.
New varieties can become
noxious weeds eg., prickly
pear.
Plant material may contain
pathogens
leading
to
epidemics.
Imported microorganisms,
or
material
containing
microorganisms eg., soil,
wood,
animal
tissues,
plant tissue, things made
from animal and plant
Unintended
spread
of
“exotic” pathogens may be
far more serious in a new
environment
due
to
absence
of
controlling
factors.
11
tissue.
Genetically
organisms
Chemicals
manipulated
Genetic manipulation may
result in new forms with
enhanced pathogenicity or
virulence.
New varieties may become
weeds or pests. New
varieties
of
microorganisms
may
become
more
serious
pathogens of plants or
animals.
Toxicological
effects,
burning of skin, fatalities
Environmental pollution
Risks are increased by:
o Inexperience
o Heavy workload
o Poor laboratory practice
o Inappropriate behaviour.
o Lack of knowledge of safety practices and procedures.
6.6 Laboratory practices to be observed when working with
microorganisms
Make sure you know the risk group and have read the SDS for that
organism.
You must submit a RAMP form to the Safety in Research and Teaching
Office (SRT) before starting work (available on the SRT website)
Dispose of all material by autoclaving at 15 Kg/cm2 for 15 min (see
section 6.15.3.1 on autoclaving).
When carrying out procedures with biological cultures these should be
done in the appropriate biosafety cabinet to prohibit inhalation of
aerosols (see section 6.14.1 on biosafety hoods).
Cultures stored in common storage areas e.g., lab fridge or cold room
must be appropriately contained and clearly labelled. Organisms that
are risk groups 3 or 4 must be stored in locked containers.
All containers and containers within containers that harbour biohazards
must be labelled with a biohazard sticker.
This includes rooms
containing biohazards, and cold storage areas containing biohazards.
6.7 Working with Genetically Modified Organisms (GMO’s)
Within Australia and its territories research on GMO’s (plants, animals,
microorganisms) is governed by a set of regulations specific by the Gene Technology
Act 2001. To administer the act the commonwealth government established the
Office of the Gene Technology Regulator (OGTR).
Organisations (academic,
industrial, military, governmental or other) must be accredited with the OGTR before
they can conduct research with GMO’s. Within each accredited organization the
regulations are administered by the Institutional Biosafety Committee (IBC). The
IBC reports to the Safety in Research and Teaching Committee (SRTC) for
administration of the regulations. More information on the IBC can be found on the
IBC website.
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The aim of the Gene Technology Act is to ensure that all work on
Genetically manipulated Organisms within Australia is carried out in such a
way to minimize threats to human health and the environment.
6.8 Work practices to be followed when doing research with GMO’s
These are in addition to the general work practices that must be followed by all
researchers section 2.
Note: Additional practices may apply if Dealings Not Intended for Release (DNIR) or
Dealings Intended for Release (DIR) licenses are in effect.
1
Every project that involves the use of GMO’s or material from GMO’s (eg. leaf
material from transgenic plants) whether made by you, or obtained from
someone else must be notified to the IBC. There are no exceptions to this
requirement. If you are not sure whether your project has been notified, ask
your supervisor, or ask the IBC. The RAMP number must be indicated on your
programme of study, or grant application cover sheet. If you do not have it
then your project has not been notified, and you will need to do this
If you need to notify the IBC information on how to do this and where to find
the application forms can be found on the IBC website.
2
Research with GMO’s must be contained within an appropriately certified
physical containment facility. A facility e.g., a laboratory or glasshouse can
only be certified by application to the OGTR through the IBC. Certified
facilities will have a (very obvious) label on the door. If you are not sure
whether your laboratory, glasshouse, growth chamber etc. is certified look at
the list of OGTR certified facilities, or ask the IBC.
If you would like to have a facility certified, ask your supervisor or the IBC.
The only exception to this requirement is for projects that fall into the exempt
category in the OGTR classification scheme. These can be carried out in noncertified facilities (although they must still be notified to the IBC).
3
You need to attend the Safety in Research and Teaching Workshop - Biosafety
run by the SRT Office. This is a requirement under the Gene Technology Act.
Details can be found on the SRT website
5
Transport of Material: If there is a need to transport hazardous biological
material, please contact the IBC. Under no circumstance should biological
material be shipped via public transport.
There are very specific requirements for the transport and storage of GMO
material out of one lab into another. Please refer to the OGTR website for
transport guideline details and ensure that you understand and follow them.
6
All material must be destroyed by incineration by an approved waste
contractor, or autoclaved using a validated autoclave cycle.
7
You must notify your supervisor and/or the IBC of any hazards, incidences, or
injuries.
8
You must keep detailed records of what GMO’s you have, how they were
made, where they are stored.
The storage containers must be clearly
13
labelled. For DNIR’s and DIR’s the OGTR can inspect at any time and request
to see the exact locations of all GMO’s covered by the license
Be Warned!
Facilities certified for work with GMO’s are inspected annually by the IBC. The aim of
the inspection is to ensure that the regulations are being complied with. The
facilities can also be inspected by the OGTR or the IBC at any time as well as on a
regular basis. Failure to comply with the regulations can lead to loss of certification
of the facility, or loss of accreditation of the organization. Penalties specified under
the criminal code can also be levied against individuals (i.e. you) and the institution.
6.9 Working with Laboratory Animals
The use of experimental animals and insects poses special problems. Animals can
harbor infectious organisms which are acquired naturally. These infections can give
rise to a chronic carrier state, or the agent might persist in a latent non-infective
form which can be reactivated periodically or as a result of certain stimuli. In 1967
the WHO Expert Committee on Zoonoses listed more than 150 diseases which affect
both man and animals. More than 50 zoonoses have been reported in Australia,
several of them being of major importance in veterinary medicine and animal
husbandry. If the possibility that such an agent may be excreted by an animal
during the course of an experiment cannot be excluded, all those animals should be
kept at a containment level appropriate to the risk.
Some experiments may involve the deliberate inoculation of animals with
microorganisms. Under these circumstances, the animal should be kept at the
containment level appropriate to the risk of the organism, recognizing that, in some
cases, in vivo work may increase that risk.
6.10 Practices for working with laboratory animals
1
In order to carry out ANY work on laboratory animals, you must first obtain
permission from the Animal Ethics Committee.
If the animal is also a GMO you must also notify the IBC.
2
The investigator and/or person(s) responsible for the animal experiment must
ensure that all those having contact with the animals and waste materials are
familiar with and aware of any special precautions and procedures that may
be required. Where possible, personnel should be protected by immunization
with appropriate vaccines.
3
It is essential that all accidents, including animal bites and scratches or cuts
from cages or other equipment, be reported and recorded.
4
Small laboratory rodents or other small animals that escape from their cages
should be killed when captured, their carcasses incinerated and the area
should be fully decontaminated. In the event that animals escape the
containment perimeter, the relevant authorities must be notified promptly and
appropriate action initiated.
5
Unexpected illness or deaths among animals must be reported to both the
14
researcher and head of animal services without delay; instructions for dealing
with such animals should be available. However, animals should not be
touched until instructions are given by the person in charge.
6
Access to the facility should be restricted to those involved in the experiment.
Be Warned!
Facilities certified for work with animals are regularly inspected by the Animal Ethics
Officer and all actions and facilities must comply with the Australian Code for the
Responsible Conduct of Research. The aim of the inspection is to ensure that the
regulations are being complied with. The facilities can also be inspected by the
RSPCA at any time. Failure to comply with the regulations can lead to loss of
certification of the facility, or loss of accreditation of the organization. Penalties
specified under the criminal code can also be levied against individuals (i.e. you) and
the institution.
6.11 Imported Biological or Biological Source Material
Importation of biological material or biological source material into Australia is
strictly controlled by the Department of Agriculture (DoA). Biological Source Material
is material that might harbor biologicals, e.g., soil or plant or animal tissue might
harbor pathogenic microorganisms.
Items such as DNA are considered to be
biological source material. You should contact the IBC if you are considering
importing biological material.
In order to import this material you must have an import permit otherwise the
material will be destroyed by customs.
1
In order to import material you must apply to the DoA for an import permit.
You can obtain an application form from the Department of Agriculture
website
2
Fill in the application form and submit to the DoA directly with the required
fee. You must submit a copy of the permit to mailto:AQIS@murdoch.edu.au
If the permit is to import quarantine material, then the material can only be
worked upon within a DoA Certified Facility -quarantine approved premises
(QAP) at Murdoch University. These are recognizable from the yellow
quarantine sticker on the door.
3
4
Within the QAP, the material must be stored within a locked container
(fridge, freezer, box within a fridge or freezer, cabinet etc).
5
A log of all incoming material, with the import permit number for each item
must be kept.
6
Records must be kept when material is removed from the quarantine store.
The name of the person removing it must be recorded.
7
If the material is to be transported to another facility, the receiving facility
must also be a QAP.
8
Transport must be in double contained shatterproof containers.
These
15
containers must not be left unaccompanied during transport.
9
Material must be destroyed by autoclaving, or incineration.
10
Records must be kept when material is destroyed.
Be Warned!!
The DoA inspect each QAP annually or more frequently if required (and we pay for
each inspection). They can pull out individual import permits and ask to see where
the material is stored and to see records pertaining to that material. In the event
that regulations governing the use of imported material are not complied with, the
import permit may be withdrawn, and Murdoch can be de-registered as a QAP as all
QAPs are linked by a single Quarantine licence. In this event no further work
involving imported material could be carried out at the facility.
6.12 Classification of Laboratories for Biological Research
The Australian/New Zealand Standard 2243.3: 2010 identifies different containment
levels of laboratories for biological research from PC1 to PC4. PC1 is the lowest level
of containment, and PC4 is the highest level. The level of containment required
depends on the material used for research. Material that present a low risk of
infection can be used in a PC1 laboratory, whereas material that presents a high risk
requires a higher level of containment (see Table 3).
Table 3: Classification of laboratories
Containment Organism OGTR Dealing
level
Risk
Group2
PC1
1
Exempt
NLRD only if lab
is certified by
OGTR
PC2
2
Exempt
NLRD3
DNIR3
PC3
3
DNIRs with risk
group
3
organisms.
PC41
4
DNIRs with risk
group
4
organisms
AQIS
Classification
QAP 5.1 For work
with
low
risk
imported material.
Facility must be
certified by Dept.
of Ag.
QAP 5.2 For work
with higher risk
imported material.
Facility must be
certified by Dept.
of Ag.
1 There are no PC4 facilities at Murdoch University. A PC3 laboratory is available in the Health
Research building (390) but is not currently certified for PC3 work.
2 Organism risk groups
3 Laboratory must be certified by the OGTR
16
6.13 How do I know if my laboratory is PC1, PC2 or PC3?
The containment level is specified by three things: (1) the physical infrastructure, (2)
the equipment in the lab; (3) the work practices in the lab. The requirements are
specified by the Australian standards 2243.3:2010. The requirements for PC1 and
PC2 containment are given in Appendix 4. A list of Murdoch facilities certified by the
OGTR and DoA is available from the IBC website.
6.14 Preventing infections
Many routine laboratory procedures generate aerosols. These are vapours consisting
of small droplets of liquid containing microorganisms. Inhalation of aerosols is a
major cause of infection by laboratory microorganisms. Aerosols frequently occur
when opening cultures of bacteria, fungi or viruses. If there is a film of liquid
between two surfaces and those surfaces are pulled apart the film is broken violently
and aerosols result. Dry material e.g. fungal spores and dried bacteria may disperse
into the atmosphere leading to inhalation if they are disturbed. There are a number
of ways we can prevent inhalation of aerosols.
6.14.1 Bio-safety cabinets
These provide a work environment that protects both the operator and/or the
material being worked on. Air is drawn into the cabinet through a HEPA (High
Efficiency Particulate Air) filter that extracts all the bacteria, fungal spores and dust.
The filtered air is then blown across the work surface protecting the material from
contamination, and is either recycled through the cabinet (protecting the operator),
or blown out of the cabinet. HEPA filters are 99.97% efficient at removing particles
of 0.3 microns from an air stream.
6.14.1.1
Class I Bio-safety cabinet
Class I cabinets provide partial personnel protection and no product protection and
are suitable for handling low-to-moderate risk biohazardous aerosols when product
protection is not essential
6.14.1.2
Class II Bio-safety cabinet
Class II cabinets provide both product and partial personnel protection and are
designed for the handling of low and moderate-risk biohazards.
6.14.1.3
Class III Bio-safety cabinet
Class III biological safety cabinets, or glove boxes, are closed-front, gas-tight boxes.
Employees work using impermeable gloves attached to cabinet-front openings or
operating ports. Class III cabinets provide the highest degree of personnel protection
and a clean work environment, and are suitable for use with highly biohazardous
agents.
Cultures of risk group 3 or 4 organisms should only ever be opened in a Class 2 or 3
BioSafety Cabinet.
17
6.14.1.4
Laminar Flow Hoods
Laminar flow hoods or clean air stations are designed to provide a flow of HEPAfiltered air from the cabinet interior over the work surface, directly towards the
operator. Toxic chemicals, allergens, infectious agents, and other potential airborne
hazards must not be handled in clean air stations because these cabinets do not
protect the operator. Use these cabinets only for procedures such as sterile filter
assembly and other jobs requiring product protection. Using clean air stations for
tissue culture preparation is not recommended because cell cultures may contain
infectious agents and allergens.
6.14.1.5
Fume Hoods
People frequently mix up biosafety cabinets and fume hoods. Fume hoods provide
protection from chemical fumes. Air is drawn into the cabinet from the laboratory,
passes over the material carrying any fumes upwards to be expelled through the
roof. They do not protect material from contamination from dust spores etc.
6.14.2 Centrifugation
Accidents leading to aerosol formation can occur during centrifugation. The number
of centrifuge accidents known to have caused infection is small; however a single
accident can involve a large number of people (one is known to have caused the
infection of 94 individuals, another 122 individuals).
The most common cause of accidents during centrifugation is breakage of tubes
leading to leakage of the contents which form an aerosol. Breakage is caused by
using inappropriate tubes, or not balancing the tubes.
Be Warned!!
Make sure you have been instructed in centrifuge use before using them. This is
especially important when using high-speed super or ultra-fuges as they are very
expensive, easy to damage, and can cause serious harm.
6.14.3 Disinfectants
Many types of chemicals can be used as disinfectants and antiseptics and there is an
ever-increasing number and variety of commercial products. Formulations must
therefore be carefully selected for specific needs, and stored, used and disposed of
as directed by the manufacturer. The germicidal activity of many chemicals is faster
and better at higher temperatures. At the same time, higher temperatures can
accelerate their evaporation and also degrade them faster. Particular care is needed
in the use and storage of such chemicals in tropical regions, where their shelf-life
may be reduced because of high ambient temperatures.
Many germicides can be harmful to humans and the environment. They should
therefore be selected, handled and disposed of with care. For personal safety,
gloves, aprons and eye protection are recommended when preparing use-dilutions of
chemical germicides. Chemical germicides are therefore not required for regular and
general cleaning of floors, walls, equipment and furniture except in cases of outbreak
control.
Chemical disinfection is often the only practical means of decontaminating work
areas and reusable materials and instruments, which cannot be autoclaved.
18
6.14.3.1
Types
Many disinfectants are available under a variety of trade names.
All have different properties and applications, advantages and disadvantages. Some
may be corrosive, toxic, flammable, explosive under certain circumstances or
possibly carcinogenic. You should familiarise yourself with the properties of the
disinfectants which you use, and read the relevant materials and safety data sheet
(SDS).
6.14.3.1.1
Chlorine
A fast-acting oxidant is a widely available and broad-spectrum germicide. It is
normally sold as bleach, an aqueous solution of sodium hypochlorite (NaOCl) and is
often combined with alkaline salts and NaOH, which can be diluted with water to
provide various concentrations of available chlorine. Chlorine, especially as bleach,
is highly alkaline and can be corrosive to metal. Its activity is considerably reduced
by organic matter (protein). Storage of stock or working solutions of bleach in open
containers, particularly at high temperatures, releases chlorine gas thus weakening
their germicidal potential. The frequency with which working solutions of bleach
should be changed depends on their starting strength, the type (e.g. with or without
a lid) and size of their containers, the frequency and nature of use, and ambient
conditions. As a general guide, solutions receiving materials with high levels of
organic matter several times a day should be changed at least daily, while those with
less frequent use may last for as long as a week. A general all-purpose laboratory
disinfectant should have a concentration of 1 g/L available chlorine. A stronger
solution, containing 5 g/L available chlorine is recommended for dealing with
biohazardous spillage and in the presence of large amounts of organic matter.
Sodium hypochlorite containing 5 g/L available chlorine is recommended as the
disinfectant of choice in emergency situations involving viruses such as Hantavirus,
and Lassa and Ebola viruses. Sodium hypochlorite solutions, as domestic bleach,
contain 50 g/l available chlorine and should therefore be diluted 1:50 or 1:10 to
obtain final concentrations of 1 g/L and 5 g/L, respectively. Industrial solutions of
bleach have a sodium hypochlorite concentration of nearly 120 g/L and must be
diluted accordingly to obtain the levels indicated above. YOU must check carefully
the concentration of available chorine at expiry date on any stock bottle as this is the
critical value.
6.14.3.1.2
Alcohols
Ethanol (ethyl alcohol, C2H5OH) and 2-propanol (isopropyl alcohol, (CH3)2CHOH)
have similar disinfectant properties. They are active against vegetative bacteria,
fungi and lipid-containing viruses but not against spores. Their action on non-lipid
viruses is variable. For highest effectiveness they should be used at concentrations of
approximately 70% (v/v) in water: higher or lower concentrations may not be as
germicidal. A major advantage of aqueous solutions of alcohols is that they do not
leave any residue on treated items.
Note. Alcohols are volatile and flammable and must not be used near open flames or
near electrical equipment. Working solutions should be stored in proper containers to
avoid the evaporation of alcohols. Alcohols may harden rubber and dissolve certain
types of glue. Proper inventory and storage of ethanol in the laboratory is very
important. Bottles with alcohol-containing solutions must be clearly labelled to avoid
their accidental autoclaving.
19
6.14.3.1.3
Iodine and iodophors
The action of these disinfectants is similar to that of chlorine, although they may be
slightly less inhibited by organic matter. Iodine can stain fabrics and environmental
surfaces and is generally unsuitable for use as a disinfectant. On the other hand,
iodophors and tinctures of iodine are good antiseptics. Polyvidoneiodine is a reliable
and safe surgical scrub and preoperative skin antiseptic. Antiseptics based on iodine
(e.g. Betadine) are generally unsuitable for use on medical/dental devices. Iodine
should not be used on aluminium or copper.
6.14.3.1.4
Quaternary ammonium compounds
Many types of quaternary ammonium compounds are used as mixtures and often in
combination with other germicides, such as alcohols. They have good activity against
vegetative bacteria and lipid containing viruses. Certain types (e.g. benzalkonium
chloride) are used as antiseptics.
Note. The germicidal activity of certain types of quaternary ammonium compounds
is considerably reduced by organic matter, water hardness and anionic detergents.
Care is therefore needed in selecting agents for pre-cleaning when quaternary
ammonium compounds are to be used for disinfection. Potentially harmful bacteria
can grow in quaternary ammonium compound solutions. Owing to low
biodegradability, these compounds may also accumulate in the environment.
6.14.3.1.5
Hydrogen peroxide and peracids
Like chlorine, hydrogen peroxide (H2O2) and peracids are strong oxidants and can be
potent broad spectrum germicides. They are also safer than chlorine to humans and
the environment. Hydrogen peroxide is supplied either as a ready-to-use 3% solution
or as a 30% aqueous solution to be diluted to 5–10 times its volume with sterilized
water. However, such 3–6% solutions of hydrogen peroxide alone are relatively slow
and limited as germicides. Products now available have other ingredients to stabilize
the hydrogen peroxide content, to accelerate its germicidal action and to make it less
corrosive. Hydrogen peroxide can be used for the decontamination of work surfaces
of laboratory benches and biosafety cabinets, and stronger solutions may be suitable
for disinfecting heat-sensitive medical/dental devices. The use of vaporized hydrogen
peroxide or peracetic acid (CH3COOOH) for the decontamination of heat-sensitive
medical/surgical devices requires specialized equipment.
Note. Hydrogen peroxide and peracids can be corrosive to metals such as
aluminium, copper, brass and zinc, and can also decolourize fabrics, hair, skin and
mucous membranes. Articles treated with them must be thoroughly rinsed before
contact with eyes and mucous membranes. They should always be stored away from
heat and protected from light.
Working solutions should be freshly prepared as organisms can grow in stored
solutions. Solutions should not be topped up and need to be labelled with name,
concentration and date of preparation.
6.14.3.1.6
Viraclean
Viraclean® passes TGA Option B and kills a broad range of other bacteria including
Enterococcus Faecalis (VRE), Acetobacter and Acinetobacter and is also proven
effective against Hepatitis B Virus, Herpes Simplex Virus and the Influenza Virus.
Viraclean® is a major development in advanced cleaning and disinfecting technology
from Whiteley Medical.
Note. Highly recommended by the IBC for most surfaces.
20
6.14.3.1.7
: F10SC
Long shelf life when diluted
Very large effective spectrum
Contains Quaternary ammonium and biguanidine compounds (5.8%), non-toxic
ampholytic surfactants and sequesterants
In Australia, F10SC is registered by the APVMA for use in animal production and
housing facilities, approved by AQIS for use in food export processing as a non-rinse
disinfectant, and is also listed by the TGA as a Hospital Grade Disinfectant.
Refer to the Reference Test Certificate Register for F10 Disinfectant.
Kills all types of pathogen – F10SC is bactericidal, virucidal, fungicidal, sporicidal
Minimal chance of microbial resistance due to F10SC’s unique benzalkonium chloride
and polyhexamethylene biguanide combination of actives and mode of action
*Rapid kill times – less than 30 secs for gram positive bacteria, 60 secs for gram
negative bacteria, Canine Parvovirus 15 mins (new testing Nov 2012)
**Successfully tested against various influenza viruses at a concentration of 1:500 in
10 mins
Non-corrosive, non-toxic, non-tainting, non-irritating, aldehyde-free
Highly cost effective
Biodegradable & ecologically friendly
Diluted solution remains active for long periods of time
Tried, tested, independently verified and documented, and approved around the
world
6.14.3.2
Activity of chemical disinfectants
This depends on:
1.
Concentration
2.
Contact time
3.
Presence of organic matter
4.
Temperature
5.
pH
6
Humidity
Make sure you use the appropriate disinfectant for your area and activity.
6.14.3.3
Treatment of Laboratory Spills
Note. For spills involving GMO’s more specific contingency plans may be required for
treating spills (please see the conditions associated with your dealing licence)
Generally a liquid spill consists of
1. The bulk of the fluid in a puddle
2. Splashes and rivulets
3. Airborne particles
Each spill is different and so each situation needs to be assessed individually.
When infected material is spilt or dropped (eg culture tube or plate) remember that
aerosols may be produced and so do not immediately bend down to clean or pick it
up but rather step back and allow time for the aerosols to settle.
As with disinfectant procedures, make sure you know how to deal with spills
appropriately in your work area.
21
6.15 Waste segregation and disposal
6.15.1 Sharps: Syringes, Needles and Blades
Sharps refer to items such as syringe needles, and blades such as razor or scalpel.
Use of these items is a common cause of injury in the laboratory. They should only
be used where there are no alternatives and must be disposed of properly. Some
areas strictly prohibit the use of sharps please check with your supervisor for area
specific restrictions on sharps use.
6.15.2 Disposal of Sharps
Needles, scalpel blades and other sharps that can easily puncture the skin should be
handled with extreme caution.
Used, disposable needles and other sharps must be placed in a rigid punctureresistant disposable container with a lid. These are readily identifiable as yellow
sharps containers in the laboratory. These containers are available from stores
around the University.
Disposable syringes with attached needles must be disposed of as one unit without
separation of the needle from the syringe.
Needles must not be re-sheathed, bent, broken, or cut.
Adapters used with evacuated tubes must not be reused. Discard needles and
adapter as a unit; do not remove needle from adapter.
Discard containers for sharps must be clearly labelled as such. Containers must also
be identified with the user’s room number.
The containers should not be overfilled as this causes needlestick injuries.
Once full, the containers are sealed and discarded into a yellow waste for incineration
bins.
Note. Sharps contaminated with radioactive material are taken to the radioactive
waste store. They are not disposed of in the waste for incineration bins. Please
refer to the Disposal of Sharps and Sharps Injury Policy
6.15.3 Decontamination and Disposal of Biologicals
6.15.3.1
Autoclaving
Most of the waste generated in biological labs can be decontaminated by autoclaving.
Autoclaving is also used to sterilize medium for growth of microorganisms, plants
and animal tissue cultures. The following factors need to be considered in
autoclaving material:
Sterilisation using steam under pressure
Correctly used, autoclaving kills bacterial spores as well as vegetative forms.

As a rule you should validate your autoclave cycle for the material you are
decontaminating. A 3L culture will take much longer to decontaminate than a
22






100ml culture. The core temperature of your material must reach the required
temperature for the required time.
Note that some materials should never be autoclaved (eg. Solvents, acids,
poisons such as phenol, chlorine, biologicals containing disinfectants or
radioactive material) as toxic or harmful substances are produced.
Steam penetration at the required temperature is necessary for successful
sterilisation so containers need to be loosely open to allow for steam penetration;
caps should be loosened on bottles, autoclave bags opened wide, bin lids
removed or not fitted tightly.
Sufficient time must be allowed for steam to penetrate the load and to reach the
desired temperature for the required time. Check with your supervisor or lab
manager for the correct timings and temperatures for sterility of your materials.
Visual indicators such as autoclave tape should be used with each load. They only
indicate if the load has reach a specified temperature (which is usually well below
the set temperature) and not how long the load has been exposed to that
temperature. The stripes on this tape turn brown when treated at 15 Kg/cm2 for
15 min. If it does not turn brown it means the material has not been successfully
autoclaved. There should be a roll of this tape in your laboratory.
Biological indicators such as spore strips (Bacillus stearothermophilus) or
chemical indicators e.g. Sterikon should be used on a regular basis (e.g.
monthly) to monitor whether loads are being sterilised effectively. The strips
should be placed in several areas of the load, including those least likely to attain
sterilisation conditions. This is generally done by staff in charge of the autoclave.
It is good practice to check for sterility of your media by incubating overnight at
370C before use, alternatively if you are concerned about exposing the media to
temperatures longer than necessary you can incubate only a small portion of the
media overnight at 370C.
Watch out for burns (particularly steam-burns), especially when opening the door
- use heat-insulating gloves when loading and unloading the autoclave.
Liquids are decontaminated by autoclaving and then disposed of down the sink
drain.
6.15.3.2
Autoclaving solid waste
Solid Waste: e.g. petri dishes etc. should be autoclaved only in autoclave bags
which have been placed in metal bins. Do not overfill bags as they are difficult to
handle and may not sterilise in a standard run despite what the bag contents may
look like after a run. Do not seal or bind the neck of the bag when autoclaving as you
need steam to penetrate the bag; the presence of moist heat is necessary for
complete destruction of many bacteria and fungi. Once autoclaved the bag is tied up
and placed into a green rubbish bin for the cleaners to remove. Ensure that you do
not autoclave any sharps in plastic bags. Solid agar cultures are placed in autoclave
bags and autoclaved. After autoclaving the bags are sealed with tape and sent for
disposal.
6.15.3.3
Autoclaving liquid waste

When autoclaving liquids in screw cap containers make sure that the lids are
not tightly screwed on otherwise the container might explode.

The container should not be full, it should be no more than 2/3 full.
 Larger volumes of liquid may require longer autoclave times, otherwise the
centre of the liquid may not be sterile.
23

On completion of the autoclave run, allow the liquids to cool before tightening
the screw caps on bottles as the bottles are not designed to withstand the
vacuum created during cooling and may implode.
6.15.3.4
Steam treatment
Steam treatment can be used to decontaminate large volumes of material where
autoclaving is impractical. An example of where this is used is:
Decontamination of potting soil in glasshouse experiments.
Decontamination of large volumes of plant material
The material can be treated in a steam box at 100-105 0C for the desired length of
time. To use this you must see the glasshouse support staff.
6.15.3.5
Incineration
Hazardous and quarantine wastes requiring incineration are sent to an external
incineration contractor (Medi-Collect) for disposal. These include infected animal
tissues and cytotoxic waste. All material for incineration should be safely bagged and
clearly labelled showing the nature of the waste, the name of the sender and the
school/area from which it comes. Infectious waste should display the biohazard
symbol.
24
7
WORKING WITH CHEMICALS AND SUBSTANCES
The risks associated with the chemicals and substances used in your project must be
assessed using the University RAMP form. In section 4 of this form, a list of
chemicals and substances is compiled and a ‘Hazardous Substances Risk Assessment
form’ (HSRA) for each is attached. Filling out the risk assessment template requires
the hazard information from the SDS form to be entered. Using the ‘consequences’
and ‘likelihood’ grid the risks associated with the chemical or substance can be
assessed. For many common chemicals there will be an existing RAF which can be
downloaded and used. Assistance is available from the Chemical Hazards and
Substances Committee to complete a HSRA for a new chemical or substance.
7.1 Know the hazards - read the Safety Data Sheets (SDS)
Before you work with a chemical you must find out the potential hazards associated
with its use. Take the time to find out how to handle them safely - it is part of your
responsibility to yourself and others under the Occupational Safety and Health
legislation. SDS are information sheets about chemicals and include information on
known or suspected hazards, incompatible chemicals and safe handling precautions.
The SDS for chemicals stored in the working area must be available in that working
area. When planning work, you must read the SDS for each chemical involved. If
the SDS for a particular chemical is not available in the area or additional information
is required, start the search with the ‘Chemwatch’ system available on the WHS
website.
7.1.1 Accessing SDS through ChemWatch
ChemWatch is an electronic database that contains information about using and
storing hazardous chemicals and substances. It can be accessed from the main
University homepage:
This should bring you to the front page of the ChemWatch program. The SDS for a
particular chemical or substance is usually required so select the ‘full’ library and
type in the name of the chemical or substance eg. lead nitrate.
25
After selecting the particular chemical from the list on the right hand side of the page
the SDS for the particular chemical or substance should appear.
7.1.2 Completing a Hazardous Substances Risk Assessment Form
(HSRA)
In some cases, the chemical required for use in your project will have a HSRA form
already in the library (note: address to this directory also privileges for this
directory). If so, you can download a copy, read it and attach it to the RAMP form.
This risk assessment has been conducted in a standard university laboratory
situation. That specifically means barrier protection of eyes with safety glasses;
covered feet and laboratory coat are assumed (internal reference here). It is also
assumed that the amounts of chemical or substance will be relatively small. Where
this is not the case for your project, the existing HSRA should be revised and
submitted.
Where there is no existing HSRA in the directory (address/reference) one must be
completed by accessing the SDS for the chemical or substance and transferring the
data into the HSRA template.
Once filled, the file is saved under the specific
chemical or substance name and attached to the RAMP form.
The HSRA template is an Excel spreadsheet file which allows users to enter data into
fields and choose from various ‘drop down’ menus. Pink fields require types input
while blue files are filled from a ‘drop down’ menu.
26
Any new HSRA are inspected by the Chemical Hazards and Substances Committee
which then makes a recommendation the WHS officer.
7.2 Labelling
The University is required to ensure that all hazardous substances are labelled
correctly. All containers, including waste containers, decanted solutions, racks of
tubes and storage boxes must be labelled if they contain hazardous substances. This
includes containers stored in -80ºC freezers, fridges, on bench spaces, in autoclave
rooms etc.
Where a substance is decanted and used immediately, no labelling is required.
However, all hazardous substances that are decanted and not used immediately
must be labelled with the product name and the risk and safety phrases. In addition
to the above, it is helpful for laboratory housekeeping if the label also contains your
name the date. Where a hazardous substance is decanted in the laboratory into
small experimental equipment, such as test tubes or columns, the label may be
attached to supporting apparatus, for example, a test tube rack.
Containers must remain correctly labelled until cleaned to the point where they no
longer contain any hazardous substance.
You are required to check all containers of hazardous substances in your laboratories
and any containers without appropriate labels need immediate action to be taken to
correctly label the container. For more details please refer to the following Policies
27
Dangerous Goods and Hazardous Substances Policy and Management of
Hazardous Substances in the Workplace Policy..
The Code of Practice ’Labelling of Workplace Substances‘, (NOHSC: 2012 (1994),
chapter 6 provides the source information for Murdoch’s guidelines and can be found
on the Safer Work Australia website:
7.2.1 Decanted chemicals/solutions
It is a legislative requirement that if you decant a hazardous substance from the
original container and it is not consumed immediately, you must label the new
container with the Product name, risk phrases and safety phrases. (All of these are
legally required to be on the original container, so just copy them across. Consult
Chemwatch if you want to look up whether or not a specific chemical is designated
hazardous. The labelling must remain on the container until it is cleaned and no trace
of hazardous material remains.
7.3 Prepared solutions/chemicals
The legislation governing the labelling of substances and solutions containing
hazardous material prepared by you is as demanding as that for the manufacturers.
You must list all of the hazardous ingredients that were used and their
concentrations in the solution. The easiest way to determine which ingredients are
hazardous and which warnings/labels they require is to look at the original
containers for the chemicals you are using.
If a component in the substance or the solution has any hazard symbols (e.g.
flammable-red diamond, oxidising agent-yellow diamond, corrosive-white diamond,
poison-white diamond, carcinogen-pink diamond or explosive-orange diamond or
harmful-orange label), place a copy of the sticker on your container. Hazard stickers
are available from the Science store.
Note that the ’carcinogen’ hazard sticker should be used for all carcinogens,
teratogens, mutagens and chemicals because the nature of the hazard is similar for
all ie. damage to DNA.
7.4 Safe storage of chemicals
The ChemWatch program contains information regarding the safe storage of
chemicals. The material (8.5.1) below is an extract from the ChemWatch entry for
lead nitrate (Pb(NO3)2) as an example.
7.4.1 Handling and Storage
7.4.1.1
Precautions for safe handling
Safe handling




Avoid personal contact and inhalation of dust, mist or vapours.
Provide adequate ventilation.
Always wear protective equipment and wash off any spillage from clothing.
Keep material away from light, heat, flammables or combustibles.
Other
information


Store in original containers.
Keep containers securely sealed as supplied.
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

Store in a cool, well ventilated area.
Keep dry.
7.4.1.2
Conditions
incompatibilities

Suitable
container
for
safe
storage,
including
any
DO NOT repack. Use containers supplied by manufacturer only.
For low viscosity materials


Drums and jerricans must be of the non-removable head type.
Where a can is to be used as an inner package, the can must have a screwed
enclosure.
Derivative of electronegative metal.
for metal nitrates:

Storage
incompatibility



Segregate from heavy metals, phosphides, sodium acetate, lead nitrate,
tartrates, trichloroethylene,
Avoid shock and heat.
Mixtures of metal nitrates with alkyl esters may explode due to the formation of
unstable alkyl nitrates.
Mixtures of a nitrate with phosphorous, tin(II) chloride and other reducing
agents may react explosively.
7.4.2 Incompatible chemicals
Even non-hazardous chemicals may become dangerous when mixed with
incompatible substances. The major incompatibilities that affect chemical storage are
listed below:



Acids and bases must be stored separately
Oxidising agents must be stored apart from other chemicals
Reducing agents must be stored apart from other chemicals
For information about particular combinations of chemicals, the individual entries in
the ChemWatch database must be assessed.
7.4.3 Storage height
Think carefully about the use of your bench storage space and make sure it is safe.
Do not store heavy items (e.g. 2L containers) on high shelves. Make sure that any
chemicals thatare either corrosive (e.g. acids, bases and oxidising agents) or
potentially damaging to eyes (e.g. acetone, acrylamide) are stored below eye level
for the shortest person in your vicinity.
7.4.4 Fridges/freezers and phenol or flammable material
Flammable chemicals with a significant vapour pressure at temperatures less than
0°C must be stored in a spark proof environment. Ordinary fridges and freezers have
29
sparking sources that can ignite the vapours. Consult the ChemWatch system
and SDS to assess whether your chemicals are suitable for fridge storage.
Organic solvents are chemicals that have appreciable vapour pressure at ambient
laboratory temperatures (15 – 35 oC).
These chemicals must be stored in
appropriate stoppered containers and within approved solvent cabinets.
7.5 Chemicals with special handling requirements
The use of some chemicals is subject to special regulation because they pose special
risks and are considered dangerous. If your project intends to use any of the
chemicals in the list below then you must have listed these on the RAMP form,
completed the risk assessment form and sought specific advice from the Chemical
Hazards and Substance committee.






benzene
carbon tetrachloride
acrylamide
hydrofluoric acid
azides
cyanides
7.5.1 Carcinogens, teratogens and mutagens
Exceptional care is required to minimize exposure for you and your co-workers to all
chemicals that fall, or are suspected of falling into these categories. Carcinogenic,
teratogenic or mutagenic chemicals must be prevented from releasing vapours or
forming aerosols and use in a fumehood with the required barrier protection is an
absolute requirement. You must get specific instruction before handling these types
of chemicals.
7.6 Compressed Gasses







Rooms where flammable gas cylinders are used should be identified by
warning notices on the doors.
Not more than one cylinder of a flammable gas should be in the room at any
one time. Spare cylinders should be stored in another building at some
distance from the laboratory. This store should be locked and identified by a
notice.
Compressed gas cylinders should be securely fixed (e.g. chained) to the wall
or a solid bench so that they are not dislodged by natural disasters.
Compressed gas cylinders and liquefied gas containers should not be placed
near to radiators, open flames or other heat sources, or sparking electrical
equipment, or in direct sunlight.
The main high pressure valve should be turned off when the cylinder is not in
use and when the room is unoccupied.
Compressed gas cylinders must be transported with their caps in place and
supported on trolleys.
Small, single-use gas cylinders must not be incinerated.
7.7 Disposal of Chemical and Substance Waste
The University conducts an annual audit of the chemicals and substances held in its
teaching and research laboratories. This audit is held to satisfy the legislated
requirement for the WHS office to maintain a list of the hazardous substances held
30
on campus. Staff charged with the responsibility of particular areas must audit
the chemicals and substances held in that area and report these to the WHS office
annually. To enable consolidation of the individual reports, these must be in a
standard format.
After this audit is completed, the University conducts an amnesty where surplus or
unwanted chemicals and substances can be disposed of without cost to the person or
group holding those chemicals. A ‘Waste Chemical Disposal Manifest’ is available
from the WSH website (https://our.murdoch.edu.au/Occupational-Safety-andHealth/Hazardous-substances/Hazardous-substances-forms/) and must accompany
chemicals and substances to be disposed of.
Outside the amnesty period, the
disposal of chemicals is charged to the person or group.
7.7.1 Organic Solvents
These are disposed of through a private company and charged back to the user.
These are disposed of through a private company and the costs charged back to the
user. It is a requirement to identify the contents of each bottle with appropriate
labelling. A Waste Chemical Disposal Manifest must be attached to each container.
The manifest requires a certain amount of information so that the external contractor
can treat the waste appropriately as well as your details so that the disposal costs
can be covered. The collection point for these waste solvents is the Physical Sciences
store (ref) and they must be deposited during store opening hours.
7.7.2 Inorganic Chemicals
Many soluble inorganic chemicals can be disposed of safely by dilution however there
are some soluble inorganic chemicals for which this is not an appropriate disposal
process. The latter category includes nearly all the ‘heavy metals’ from row 4
onwards in the periodic table. Insoluble inorganic chemicals must be disposed of as
solid residues using the ‘Waste Chemical Disposal Manifest’ available from the WSH
website. Advice is available from the Chemical Hazards and Substances Committee.
7.7.3 Organic Chemicals
Many water soluble organic chemicals can be disposed of safely by dilution because
they are readily bio-degradable.
Non polymeric chemicals with relatively low
molecular weight usually have very short half-lives in waste treatment systems and
can be safely diluted. There are chemical degradation strategies available to deal
with organic chemicals which resist bio-degradation. Organic chemicals that resist
these treatments can be disposed of by incineration. This ultimate disposal strategy
is conducted by an external contractor and a ‘Waste Chemical Disposal Manifest’ is
required.
It is impossible to make many generalisations about safely disposing of chemicals.
Fortunately most chemicals and the risks they pose to the environment are well
known. This information is contained in the Material Safety Data Sheets (SDS) that
can be viewed and copied from the ChemWatch data base. In assessing disposal
risks the precautionary principle must be observed, particularly when the chemical or
substance is not common or well known. Advice is available from the Chemical
Hazards and Substances Committee (CHSC) and the WHS office.
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8
WORKING WITH RADIOISOTOPE
Very specific requirements surround the use of sealed and unsealed radioactive
sources. You must contact the Radiation Safety Officer before commencing any
work in this area.
If you plan to work with radioisotopes you should ensure that you enrol for the UWA
“Unsealed Isotope Handling Course” before you begin. The course is only run twice a
year and has been made mandatory for Murdoch University staff and students
working with isotopes.
8.1 Individual User Responsibilities:







Each individual is responsible for maintaining his/her exposure As Low As
Reasonably Achievable (ALARA).
All users must adhere to proper handling and radiation safety procedures
when working with radioactive materials. Each individual who works with
radioisotopes is responsible for the proper disposal of radioactive waste,
maintenance of records of all disposal and proper identification of radioactive
materials and identification of contaminated equipment with URSO approved
labels.
The individual user is responsible for being familiar with radiation survey
procedures and checking work areas for contamination periodically or after
each radioisotope procedure. He/she is also responsible for checking hands,
body and clothing for radioactivity and removing contamination before leaving
the laboratory.
The individual user is responsible for the proper use and handling of his/her
personnel monitoring badge, furnishing bioassay samples to the Radiation
Safety Office for analysis when requested and obtaining periodic thyroid
uptake measurements when working with radioiodine as applicable.
Each individual who works with radioisotopes must be familiar with the basic
elements of decontamination procedures and is responsible for the cleanup of
contamination for which he/she is responsible.
All users should be familiar with the characteristics of the radioisotopes they
are working with.
Incidents involving contamination of personnel, uncontained spills, theft or
loss of radioactive material and suspected overexposure must be reported
immediately to the Radiation Safety Office.
8.2 Radioactive Substances Laboratory Guidelines and Procedures
8.2.1 Who can work in the laboratory?



Access to laboratory is not permitted without prior permission of technical
staff in charge of the lab or Radiation Safety Officer.
Work within the laboratory must be conducted only by persons with a current
radiation license, or by someone working under the license of somebody else.
All workers must have completed the UWA radiation safety course.
License holder must ensure the people who work under them are doing so in
a safe manner that complies with all regulations and declared safety
standards.
8.2.2 Bringing radioactive samples to the lab and storing them:

Any radioactive sample you wish to bring into lab must first be authorised.
This requires a form to be filled out prior to the sample being received or
32


sent. This form is available from technical staff and permission will then be
given or refused to bring in the sample when the form is returned.
How and where the sample is to be stored within the lab will be discussed
with relevant technical staff before the sample is received. Samples not being
immediately used must be stored appropriately at all times.
Documentation of each sample is to be kept on file within the lab along with
the form authorising the sample to be used within the lab
8.2.3 Radiation monitoring:





It is advised that a log book is maintained in the lab and completed daily by
all staff and students working in the lab.
Each worker is required to wear a radiation monitor at all times in the lab This
is to be handed in to technical staff once a month for results to be
downloaded and checked to ensure that received doses are within safety
limits.
Once a fortnight the lab space is to be checked using a hand scanner and
readings entered on a map of the lab along with the date. A copy of all checks
will be kept on file within the lab.
Daily spot checking of spaces where radioactive substances have been
handled is expected.
Any items that are being removed from the labmust first be cleaned and
checked using a hand held radiation monitor to ensure they are not
contaminated.
8.2.4 Disposal of radioactive substances:


Radioactive contaminated tissues, paper towel, and other general waste lab
items that would normally go in a bin must be disposed of into a plastic lined
cardboard radioactive waste box. These are available from the science store.
The details of the contents must be recorded on the front of the box when it
is first used, and activities measured and recorded on the box once it is full
and sealed up. Boxes are then given to technical staff who will arrange
disposal.
At the end of a project any unused or remaining radioactive material and
waste must be returned to its original source or disposed of correctly.
Abandoning radioactive material for others to deal with is not an option. You
will be required to declare on your form requesting permission to bring a
sample into the lab how and where the sample will be disposed of.
8.2.5 General laboratory procedures



Each worker in the lab will be assigned a workspace and possibly a storage
space as well. It is that workers responsibility to keep their spaces tidy and
safe. This involves clearing away at the end of each shift, returning samples
to appropriate storage, disposal of chemicals after use, and taking a share of
responsibility for condition of common areas such as floors, shared benches,
fume cupboards and balances. Technical staff may periodically check
workspaces and storage spaces to ensure they are being kept in a satisfactory
manner.
All samples must be clearly marked with a chemically meaningful name,
radioactive species, and name of user. Original sample containers must
include as much information as possible, including name of user’s supervisor.
Lab coats must be used only in the designated lab and not removed from the
lab. Please consult with technical staff for instructions on how to clean your
lab coat.
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9
FIELDWORK
Staff and students who engage in fieldwork as an extension of, or in association with,
laboratory work need to address all issues and arrangements associated with the
safe conduct of fieldwork. Fieldwork is essentially work conducted outdoors on
campus or in other locations, in other institutions, or overseas. Depending on the
location and the type of research or teaching being undertaken in the field, the
safety risks involved may vary enormously. These risks need to be considered and
planned for. The information below will facilitate this process.
The risks associated with the conduct of fieldwork must be assessed using the
Murdoch University RAMP form. This form will enable hazard identification. In
addition, completion of Fieldwork Risks will identify if there is a need to complete a
Murdoch University Fieldwork Safety form. The Fieldwork Safety Form requires
information pertaining to the following areas:
 The nature of the fieldwork activity (including work with fauna and flora)
 Participants involved and their training and expertise
 Equipment, vehicles and tools required
 Required permissions, certification and licenses
 Fieldwork locations, travel and transport, and communication
 Where relevant, information will also be collected concerning health, first aid,
clothing and safety equipment.
The Fieldwork Safety Form is a mechanism designed to, not only identify risks, but to
assist in the creation of a Fieldwork Safety Plan. This must be lodged in the relevant
School, and approval must be obtained, at least two weeks prior to the conduct of
fieldwork.
Note. Fieldwork cannot be undertaken without a RAMP number and compliance
with all identified necessary approvals.
34
Full details of the various safety committees supporting this documentation
can be found on the SRT website