Download An Introduction to Biosafety

An Introduction to Biosafety
Covering the basic principles of
containment and working safely
when handling biological materials
Phil Walsh and Fiona Harris
Biological Safety Advisers
Biological Safety Unit
Health and Safety Department
Information and Reference documents
Biosafety Unit website:
http://www.ed.ac.uk/schools-departments/healthsafety/biosafety/home
Various sections:
General, Containment Laboratories, Pathogens,
Genetic modification, Work with other materials,
Risk assessment forms, Transport
In particular read:
 Good microbiological practice and containment
 Hazard grouping and containment requirements
 Preventing injuries from glass and sharps
 Requirements under the GMO (Contained Use)
Regulations
Select others depending on nature of your work
Safety cabinets, Work with blood and tissues, Work with
naked DNA, Work with animals
Individual workers responsibilities
 familiarise yourself with University and School Health and
Safety Policies
 carry out your work safely and in accordance with these
policies using all control measures specified
 exercise care in relation to yourself and others who may
be affected by your actions (or inactions)
 report any safety concerns
Legislation
Also: the Plant Health (Great Britain) Order 1995 (as amended)
Advisory Committee on Dangerous Pathogens
Legislation
Approved Guidance
Biological Agent
• Biological agents are included within the COSHH
definition of a ‘substance hazardous to health’
• Biological agent defined as:
micro-organism, cell culture or human endoparasite,
whether or not genetically modified, which may cause
infection, allergy or toxicity
or otherwise create a hazard to human health
Biological agents
Micro-organisms
bacteria
viruses
fungi
prions
Cell Culture
Endoparasites
Adventitious agents
Microscopic endoparasites
e.g. malarial parasites,
amoebae & trypanosomes
Microscopic infectious
forms of larger parasites
e.g. microscopic ova and
infectious larval forms of
helminths
Duties under the COSHH: Identify hazards and
assess risks to human H&S
• identify and classify biological agents which will be used
• consider all hazards not just infection
• first consider substitution with less hazardous agent or alternative
method of work or work process
• take the nature of the work into account and the route of infection
of the particular agents involved
• identify any particular groups of people who may be at increased
risk
Control the risk
•
prevent exposure wherever possible
•
if not reasonably practicable then “adequately control”
• maintain, examine and test control measures
• monitor exposure
• provide information, instruction and training
• provide health surveillance
• make arrangements to deal with accidents & emergencies
Schedule 3 COSHH: biological agents
• maintain list of employees working with the most hazardous
biological agents
• notify use and storage of certain agents
• notify consignment of certain agents
Once systems are in place you must
monitor
• to ensure control measures are working effectively
periodically review
• to ensure these are still valid and applicable
Also take account of any new information that becomes
available that affects the risk assessment
Risk assessment considerations
• Hazards of the biological materials
• Routes of infection
• Control measures
Hazards of biological materials
• Infection
• Toxic
• Allergenic
• Carcinogenic
• Other?
Routes of infection
• Inhalation
breathed in
• Ingestion
by mouth
• Injection
damaged to skin
• Absorption
across mucous membranes e.g. splash into eye
Basis for control measures
Block all routes of infection:
• Do not put anything in mouth
• Do not create aerosols
• Cover damaged skin and minimise risk of puncture
wounds
• Do not produce splashes and droplets
• Do not transfer to any route with hands or other
contaminated objects
Hazard Group classification system
ACDP classifies biological agents which may be a hazard for
human health into four Hazard groups based on:
A. Ability of the agent to cause infection in healthy humans
B. Severity of the disease that may result
C. Risk that the infection will spread to the community
D. Availability of effective treatment or prophylaxis
Hazard Group matrix
Approved List of Biological Agents
• ACDP classifies biological agents which may be a hazard for
human health into Hazard groups 2-4
• Hazard Group 1 agents are not pathogenic but may still cause
opportunistic infections
• The ACDP classification is approved by the HSE → published as
the Approved List of Biological Agents
HG1
Disabled E. coli
HG2
Listeria
monocytogenes
HG3
Bacillus anthracis
HG4
Ebola virus
• Agents not on the Approved List: must seek evidence of
pathogenicity- not automatically assign to Hazard Group 1
• all unlisted viruses isolated from humans assigned to not less
than Hazard Group 2 - unless evidence that they are not
pathogenic
• if there is any uncertainty then the higher of the two possible
hazard groups should be chosen
• GMMs are not on the Approved List but the wild-type species
may be listed
Other considerations in the risk assessment
• Hazard Group does not take account of other hazardous
properties e.g. toxic or allergenic
• Categorisation is based on the infective hazard to healthy
adult workers
• It does not allow for any additional risk to individual workers
caused by for example
•
•
•
•
pre-existing disease
the effects of medication
compromised immunity
pregnancy or breastfeeding
Containment levels
• Working safely with biological agents requires a series of
containment measures termed Containment Levels
• Four containment levels directly related to each hazard group
Hazard Group number = Containment Level number
• Laboratories where known biological agents are propagated or
concentrated → match hazard group of the agent with the CL
e.g. HG3 pathogen, Bacillus anthracis = CL3
Containment levels
At each CL, control measures are specified in relation to:
• Facilities
• Procedures - local laboratory rules & Code of Practice
High Specification laboratory
Strict Codes of Practice
CL4
High risk
microbes
CL3
CL2
Basic laboratory
GMP
CL1
Low risk
microbes
Containment Level 1
Facilities
• Easy to clean laboratory
• Impervious and resistant benches
• Wash basin or sink for hand washing
• Appropriate warning signs
Containment Level 1
Working practices
• No eating, chewing, drinking, smoking, applying makeup
• No mouth pipetting
• Wear lab coat and remove when leaving lab
• Wash hands
• Procedures for use, storage & transporting biological
agents
• Procedures for disinfection & waste disposal
• Procedures for accidents & incidents
Containment Level 2
all the requirements of CL1 plus
Facilities
• Restricted access
• Dedicated hand wash basin at exit
• Adequate space for safe working
• Separate storage area for lab coats in the lab
• Negative pressure if mechanically ventilated
• Microbiological safety cabinet may be required if an infectious
aerosol is produced
Containment Level 2
all the requirements of CL1 plus
Working practices
• Same basic working practices but applied to a higher standard
• Greater level of information, instruction, training & supervision
required
• Workers to receive training in handling pathogens
Containment Level 2
Additional control measures and precautions
• Extras depending on nature of work and route of infection
•
Avoid using sharps
•
Gloves
•
Microbiological safety cabinet
•
Designated work station
•
Eye protection
•
Vaccination
Containment Level 3
all the requirements of CL2 plus
• Facilities
•
Self contained facility
•
Separated from other activities
•
Operate at negative pressure
•
HEPA filter on extract
•
Sealable for fumigation
•
Microbiological safety cabinet (ducted)
•
Viewing panel
Containment Level 3
all the requirements of CL2 plus
Working practices
• High standard of training and supervision
• All working practices rigorously controlled, documented &
recorded
• Use dedicated lab coats and autoclave before laundering
• When handling infectious materials wear gloves and carry
out all manipulations in MSC
• No sharps
Containment Level 4
• Totally isolated
• Complete change of clothes
• Shower out
• Sterilise all waste and effluent
• Double ended autoclave
• Very strict air handling arrangements
Classification
ACDP hazard
group
1-4
Pathogenicity
Containment
level
1-4
Facility &
Working Practices
SACGM
classes
1-4
Project
MSC classes
I, II, III
Type of cabinet
Simple system for implementation of risk
control measures
1. identify biological agents to be used or those that may be
present in the materials to be handled
2. classify each into one of four Hazard Groups
3. the number of the Hazard Group indicates the level of
containment under which the work must be carried out
4. identify whether any additional control measures needed
Steps in a COSHH risk assessment
Use form BA1 (revised 2003 ver2)
Available from the biosafety unit website
Assessing risk for work with blood & human
tissues
hazards

blood borne viruses

hepatitis viruses, HIV

other pathogens
vary depending on

type of material

source of material
Control measures for work with blood
and human tissues
 Containment Level 2
 Gloves
 Avoid use of glass and sharps
 Designated work area
Guidance on work with blood and human tissues on biosafety
unit website
Range of Guidance available
on the biosafety unit website
 Blood and human tissues
 Tissue culture
 Naked DNA including oncogenes and viral genomes
 TSEs
 Animals
 Plants
Separate regulations for GM work
The Genetically Modified Organisms (Contained Use)
Regulations 2014
Specific requirements for risk assessments

need approval from the GM safety committee

project is classified as Class 1- 4

notification to HSE
Control measures on same principles as for all biological
work
Microbiological Safety Cabinets
An introduction to selection
and safe use of MSC
Information and Reference documents
Biosafety Unit website:
http://www.ed.ac.uk/schools-departments/healthsafety/biosafety/home
Guidance

Microbiological Safety Cabinets
Microbiological Safety Cabinets
 Offer protection to the user and the environment from
infectious aerosol hazards
 Local Exhaust Ventilation (LEV) under COSHH
regulations
 Specification in British Standard BS EN 12469 (2000)
3 different types
 Class I
 Class II
 Class III
 All protect worker
 Some also protect work
 All exhaust air is HEPA filtered
Class I
Class II
Class III
Do not confuse with

Laminar flow hoods

Tissue culture cabinets

Anaerobic cabinets

Fume cupboards
Safe use of MSCs
 All users must be trained in the correct use of MSC
 Do not use until the cabinet has warmed up
 Do not work with the UV light on
 Do not obstruct the air intake grilles in Class II cabinets
 Do not use bunsen burners or centrifuges in Class II
cabinets
 Always disinfect the cabinet after use
Cabinet selection
 Select the correct MSC for your work (by risk assessment)
 Consider whether protection of the work is needed
 MSC class is not linked to the CL assigned to the work
Class
Operator
Protection
Suitable for which
biological agents
Protects work from
contamination
I
Yes
HG2 & HG3
no
II
Yes
HG2 & HG3*
yes
III
Yes
HG3 & HG4
yes
* Risk assessment required
Key Points
 A cabinet only offers protection against infectious
aerosol hazards
 The level of protection offered by a cabinet relies
heavily on good working practices
Thank you!
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