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Biological and GM safety

Occupational Health

& Safety Service

Welcome & Introduction Occupational Health

& Safety Service

Samantha Dainty – Biological Safety Adviser

AgendaBiosafety and Biosecurity

• Legislation relating to biosafety • Biosafety and Biosecurity Management Standard• Classifying biological agents• Risk assessment• Notifications and licensing• Containment and controls• Emergency Planning

Genetic Modification• GM Management Standard• GM risk assessment and peer review process

Biological Agents (COSHH) Occupational Health

& Safety Service

Micro-organismsHuman endoparasites

Human and animal samples

Environmental samples

Toxins

Allergens

Other legislation Occupational Health

& Safety Service

SAPO

zoonoses

Schedule 5

Plant health orderGM

animal pathogens

Biosafety and Biosecurity

Management Standard Occupational Health

& Safety Service

• How we will comply with Biosafety legislation

• Who is responsible for putting the standards into action

• Applies to everyone at all levels

Occupational Health

& Safety Service

The standard (duty)

How to meet the standard Who’s doing

what

Biosafety and Biosecurity

Management Standard

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Classification of Biological Agents

COSHHOccupational Health

& Safety Service

Human pathogens

Group Description Examples

HG1 Unlikely to cause human disease B. subtilis, E. coli (K12 and BL21 strains)

HG2*

Can cause human disease and may be a hazard to employees

Unlikely to spread to community

Usually effective prophylaxis or

treatment

Influenza virus, Listeria Monocytogenes, Adenovirus,, E. coli spp, S. aureus

HG3*

Can cause severe human disease and may be a serious hazard to employees

May spread to community

Often effective prophylaxis or treatment

Pandemic influenza virus, HIV, HBV, HCV, E. coli 0157, M. tuberculosis,

HG4

Causes severe human disease and is a serious hazard to employees

Likely to spread to community

Often no effective prophylaxis or treatment

Pandemic influenza virus, Variola virus, Ebola virus

Or self-classify

Classification of Biological Agents

SAPOOccupational Health

& Safety Service

Animal pathogens or zoonoses

Group Description Examples

SAPO1Disease-producing organisms which are enzootic (native in animals in this country) and do not produce notifiable disease.

SAPO2

Disease-producing organisms which are either exotic or produce notifiable disease, but have a low risk of spread from the laboratory.

Trypanosoma Bruciae, Bovine leucosis virus

SAPO3

Disease-producing organisms which are either exotic or produce notifiable disease and have a moderate risk of spread from the laboratory.

Bacillus anthracis, Vesicular stomatitis virus

SAPO4

Disease-producing organisms which are either exotic or produce notifiable disease and have a high risk of spread from the laboratory.

Avian influenza virus, Foot and mouth disease virus, Rabies virus

Appendix 1 (Schedule 1 of SAPO)

Classification of Biological Agents

Plant Health OrderOccupational Health

& Safety Service

Plant Health (England) Order

Group Description

Schedule 1 Plant pests which shall not be landed in or spread within England.

Schedule 2Relevant material which may not be landed in or moved within England

if that material is carrying or infected with plant pests

Schedule 3Relevant material which may not be landed in England if that material

originates in certain third countries

Schedule 4Restrictions on the landing in and movement within England of relevant

material

Schedule 5 Relevant material from a third country for which a phytosanitary

certificate may be required

Schedule 6Prohibitions on the landing in and movement within England of relevant

material without a plant passport

Schedule 8Swiss plant passports

Schedules 1-6 & 8

Classification of Biological Agents

Schedule 5Occupational Health

& Safety Service

• Intact Schedule 5 micro-organism• Genetically modified but retain the ability to cause serious harm to

health• Nucleic acid derived from a Schedule 5 micro-organism that can encode

infectious and/or replication competent forms of the micro-organism• Nucleic acid sequence derived from a Schedule 5 microorganism which

when inserted into any other living organism alters or enhances that organisms's ability to cause serous harm to human or animal health

• any quantity of botulinum or Clostridium perfringens enterotoxin/epsilon toxin

• more than 5 mg of any other schedule 5 toxin• Any nucleic acid sequence coding for a Schedule 5 toxin or a genetically

modified micro-organism containing such a sequence

Notifications and LicensingOccupational Health

& Safety Service

Pathogen and toxin registration

Plant Health OrderControlled or prohibited plants,

plant products or plant pests

under Schedules 1-6

Schedule 5Anti terrorism, Crime & Security

Act

Specified Animal

Pathogen

1 2 3 4

ACDP Hazard Group

1 2 3 4

Materials that may

contain biological

agents

Risk AssessmentCL1

CL2

CL3

CL4

CL2

CL3

CL4

Compliance with the conditions of the licence

See OHSS H&S Guidance 301.3

Biosecurity policy in consultation with CTSA

Pathogen registration

Pathogen registration

Pathogen registrationPathogen registration if known/very likely to contain ACDP HG2-4, SAPO

2-4 etc.Pathogen registration

HSE notification

*Only 1st

use

SAPO/IAPO licences

Plant Health licenceNotifications or licences depending

on nature of the sample CTSA notification

Classification

Assessment

Containment

Registration

Notification

Requirements for work with biological agents according to their classification

HG3, SAPO3, PHO licences, Schedule 5 = Peer review by Biosafety Committee

AND Let us know when your work has ceased

BioCOSHH Risk AssessmentOccupational Health

& Safety Service

Review every 2 years (at least)

Advice:• Biological Safety

Supervisors• School Safety

Officers• Biological Safety

Advisor

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Routes of exposure Occupational Health

& Safety Service

Mucous Membranes:Exposure of mucous membranes of theeyes, nose and mouth through splashes,splatter or aerosols.

Inhalation:Breathing in respirable sizedaerosols (<5μm), centrifuge leaks,spills, pipetting, etc.

Ingestion:Swallowing through mouth pipetting, eating, drinking orsmoking in the lab.

Percutaneous/injection:Through intact or non-intact skin viaNEEDLESTICK, puncture with contaminated sharp object, animal scratch or bite, through wounds abrasions, or eczema.

Contact (indirect transmission):Via mucous membranes or non-intact skin from hands that have been in contact with a contaminated surface(i.e. benches, phones, computers, equipment handles) or by failure to wash hands after working

Infectious Dose Occupational Health

& Safety Service

Agent Dose Route

Tuberculosis 10 Inhalation

Influenza A2 790 Inhalation

S.typhi 105 Ingestion

Vibrio cholerae Cholera 108 Ingestion

E.coli 108 Ingestion

E.coli 0157 10-100 Ingestion

Shigella 10 Ingestion

Polio virus 1 2 Ingestion

Amount of pathogenic organisms that will cause infection in susceptible subjects

Quantity and Concentration Occupational Health

& Safety Service

Highly concentrated substances can be:• More infectious• Higher aerosol potential• Emergencies require specialist

spill clean up • Waste inactivation issues

• Higher risk

Small scale

Medium scale Large scale

Concentration, centrifugation, filtration

Who is at risk? Occupational Health

& Safety Service

Experience –impacts likelihood of exposure

Shared spaces –who else needs to be informed?

Anyone at increased risk?- Immunocompromised- Asthma- Dermatitis Severity of exposure

Higher risk? Supervision/access control

Risk Estimation Occupational Health

& Safety Service

Nature of organism/hazard group

concentration

Route of exposure

FrequencyInfectious dose

Controls – Containment labs Occupational Health

& Safety Service

Containment level 1 - hazard group 1 Containment level 2 - hazard group 2 Containment level 3 - hazard group 3

Containment level 2 for when there is no intention to deliberately propagate or concentrate a biological agent but there are uncertainties about the presence of HG2, HG3 or HG4 in the sample.

Containment level 3 or 4 where appropriate if it known or suspected that such a level is necessary even if there is no intention to deliberately propagate a HG3 or HG4 biological agent

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Controls – Containment labs Occupational Health

& Safety Service

Facilities

Equipment

System of work

Waste and disinfection

Animal containment

Containment level 1 Occupational Health

& Safety Service

• Hand wash sinks

• Bench surfaces (and floors) impervious to water and chemicals. Easy to clean

• Autoclave on site (GM)

• Suitable protective clothing

• Specified disinfection procedures

• Inactivation of GMMs by a validated means

• Written records of staff training

Containment level 2 Occupational Health

& Safety Service

• Procedures involving infectious aerosols must be contained within an MSC (or similar)

• Access restriction

• Biohazard sign on door

• Autoclave in the building

• Inactivation of all biological materials by a validated means

• Safe storage of biological materials

Containment level 3 Occupational Health

& Safety Service

• Isolation of laboratory suite

• Lab sealable for fumigation

• Entry via airlock

• Negative pressure

• HEPA filtered extract

• Procedures with infectious materials must be contained within MSC or similar

• Access restriction

• Biohazard sign on door

• Autoclave in the lab

• Inactivation of all biological materials by a validated means

• Safe storage of biological materials

SAPO Containment labs Occupational Health

& Safety Service

Containment level 2 – SAPO group 2 Containment level 3 – SAPO group 3

Containment labs Occupational Health

& Safety Service

Plant Health Licences – You must meet the conditions of the licence

Schedule 5 – Security control measures as advised by CTSA and a Biosecurity Policy

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Multiuser labs Occupational Health

& Safety Service

• All users must comply with the requirements of the lab containment level regardless of the hazard group of the organism they are working on

• Good communication required

Controls – COSHH Hierarchy Occupational Health

& Safety Service

Hierarchy of control selection

Attenuated strainsAuxotrophs‘lab’ strains

Microbiological Safety CabinetsOccupational Health

& Safety Service

Microbiological Safety CabinetsOccupational Health

& Safety Service

Correct Operation is essential for containment

• Do not obstruct grill

• Bunsen burners, centrifuges and walk-past all disrupt air flow

• MSCs do not sufficiently vent harmful chemicals. Avoid volatile chemicals –methanol, acetic acid, mercaptoethanol

• Caustic material that vaporize can attack aluminum separators in the HEPA filter

• Statutory requirement – annual LEV testing

Containing aerosols in centrifugesOccupational Health

& Safety Service

• Use biosafety rotors for biological agents (CL2+)

• Open rotor, buckets, tubes inside MSC

• Do not open centrifuge immediately after a breakage/spillage – let aerosols settle first

• Disinfect after spillages

Animal Containment Occupational Health

& Safety Service

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Administrative Controls Occupational Health

& Safety Service

training Access control

SignsSystems of work

Personal Protective Equipment Occupational Health

& Safety Service

Lab coats

Gloves – select the right type

Eye protection

Respiratory protection – face fit testing

Immunisation Occupational Health

& Safety Service

Area Activity ** Required Recommend

Clinical Patients/clinical materials contact – exposure prone procedures Hep B*

Patients/clinical materials contact – non exposure prone procedures Hep B

Laboratory Working with unscreened human blood or tissue Hep B

Working with screened human blood or tissue Hep B

Working with novel human cell lines from uncontrolled sources Hep B

Working with established human cell lines from controlled sources

NB Has the cell line been screened? Pre 2010 ATCC?

Hep B

Working with non human material

Agricultural /

Horticultural

Handling soil or plant material Tetanus

Animal

Technicians

Working with colony bred animals Tetanus

Working with wild caught primates if not conditioned Rabies, Hep A Tetanus

Health Surveillance Occupational Health

& Safety Service

• We are required by law to keep a list of those working with hazard group 3 biological agents

• Asthmagens

• Allergens

• Certain pathogens e.g. TB

Health surveillance is required if all the following criteria are met:

• There is an identifiable disease/adverse health effect and evidence of a link with workplace exposure

• It is possible the disease/health effect may occur• There are valid techniques for detecting early signs of the disease/health

effect• These techniques do not pose a risk to employees

Transport of Biological Samples Occupational Health

& Safety Service

• Between labs - Secondary containment• Between university sites:

• Between Organisations – Sign up for the training course!

Principals of Good Microbiological PracticeOccupational Health

& Safety Service

Aseptic technique

Aerosol minimising techniques

housekeeping

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Waste Inactivation - autoclaving Occupational Health

& Safety Service

• Waste must be inactivated by a validated means – autoclave in preference to disinfection

• CL2 – before removal from the site

• CL3 – before removal from the facility

• Applies to ALL GM samples

• But…..do not autoclave samples containing radioactive or hazardous chemicals

monitoring

Waste Inactivation - autoclaving Occupational Health

& Safety Service

Contaminated with biological agents or GM. No chemicals (residues OK)

Contaminated with biological agents or GM. Can’t be autoclaved

Not contaminated with biological agents but looks like lab waste (e.g. gloves from a chemistry lab) No chemicals

Energy from waste/deep landfill

High temperature incineration

Take the tops off your tubes!!!

Sharps Occupational Health

& Safety Service

Go

od

Pra

ctic

eG

oo

d P

ract

ice

Don’t dispose of sharps in ordinary waste bins

Don’t dispose of sharps in clinical waste bags

Never resheathneedles

Use gloves and never resheathneedles

Don’t transfer used sharps to other workers

Dispose of sharps immediately after use in a sharps bin

Take disposal bin to sharps

Dispose of bins on reaching max level (marked)

Treat all contaminated sharps as hazardous

Bad

Pra

ctic

e

Waste Inactivation - disinfectants Occupational Health

& Safety Service

Disinfectant Bacteria Bacterial

spores

Fungi Enveloped

viruses

Non-enveloped

viruses

Prions Comments

Phenolic YES NO YES YES Limited NO Toxic

Hypochlorites YES YES Limited YES YES YES Toxic/Corrosive

70% Alcohol YES NO NO YES Limited NO Flammable

Aldehydes* YES YES YES YES YES NO

Irritant/allergen,

glutaraldhyde resistance

bacteria

*Formaldehyde YES YES YES YES YES NOCL3 level - very toxic,

need to seal the lab!

Peroxygen

(Virkon)YES YES YES YES YES NO

Dust irritant, limited

solution life, corrosive to

metals

A specified disinfection procedure is required for all work with biological agents and GM microorganisms

Waste Inactivation - disinfectants Occupational Health

& Safety Service

Validation!

How do you know if disinfection has been

successful?

• data from manufacturer – kills 100% of all

known germs!

• data from other users / labs

• ‘in-use’ testing

Waste Inactivation - disinfectants Occupational Health

& Safety Service

• some disinfectants can be ‘inactivated’ under different conditions

• presence of ‘organic material’ -compensate

• avoid disinfectant combinations

• limit age of disinfectant solution (fresh)

• use at correct dilution (‘in-use’ concentration)

• exposure time

• COSHH properties; compatibility with hardware

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Emergency Planning Occupational Health

& Safety Service

Spills

autoclave bags

gloves

Mask (HEPA filtered)

lab coat

absorbent towels

disinfectant

goggles

absorbantgranules

SOP

gloves (heavy duty)

sealable bin(autoclavable)

biohazard tape

overshoes

Emergency Planning Occupational Health

& Safety Service

Needlestick/Contaminated Sharps

• Action after exposure to blood borne viruses – Occupational Health

• Risk assessment

• Procedures for potential exposure to HepB (if unvaccinated), HepC and HIV

• counselling

Genetic Modification

Occupational Health

& Safety Service

Definition Occupational Health

& Safety Service

Any technique which alters the genetic material of an organism using a method that would not occur by natural mating or recombination

Gene deletions or insertion of multiple copies of a gene if brought about by artificial means

e.g. Insertion of nucleic acid, produced outside an

organism into any virus, plasmid or other vector then

incorporation into a host organism in which it does not

occur naturally and a capable of continued propagation.

e.g.. Direct introduction into an organism of heritable

genetic material prepared outside the organism,

including micro-injection and micro-encapsulation.

Exemptions Occupational Health

& Safety Service

Mutagenesis (eg x-rays, chemicals)

Synthetic nucleotides

Self cloning organisms

“Natural” transformation

Hybridoma’s

Humans and human embryos - IVF

GM risks: The Unknown Occupational Health

& Safety Service

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GM risks: Public Perception Occupational Health

& Safety Service

We have to protect ourselves/work from this controversy by demonstrating

the highest level of containment of GMO – “zero risk” of escape……

Genetic Modification Management

Standard Occupational Health

& Safety Service

• How we will comply with the Contained Use regulations

• Who is responsible for putting the standards into action

• Applies to everyone at all levels

Risk Assessment Occupational Health

& Safety Service Risk assessment for……. Occupational Health

& Safety Service

Gene cDNA

Host

Vector

Geneticmaterial

GM cells

Use this to help! Occupational Health

& Safety Service

Health and safety inspectors seek to secure compliance with the law and may refer to this guidance as illustrating good practice

Waste Inactivation Occupational Health

& Safety Service

All GM waste must be inactivated by a validated

means

Autoclaves in preference to disinfectants

Must be described in the GM risk assessment

Any deviation from this must be in discussion with

the Biological Safety Adviser - Consultation with

HSE

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Controls and Activity Class Occupational Health

& Safety Service

Containment necessary to control

the risk

Risk Classification

Level 1 Class 1

Level 1 with the addition of measures from level 2

Or

Level 2 (without additional measures)Class 2

Level 2 with the addition of measures

from Level 3

Or

Level 3 (without additional measures)

Class 3

Level 3 with the addition of measures

from Level 4

Or

Level 4 (with or without additional

measures)

Class 4

Control Selection and Activity Class Occupational Health

& Safety Service

HG1 - GM Class 1 – CL1

HG2 - GM Class 2 – CL2

HG3 - GM Class 3 – CL3

BUT an HG1 organism could become a GM class 2 depending on how it’s been modified

Hazard group rating sets the “base level” then depending on the modification the organism may be elevated to a higher risk group based on the modification

Control Selection and Activity Class Occupational Health

& Safety Service

Class Criteria for Classification1

All criteria must be

met to justify the

assignment of

Class 1

3rd generation (or safer) vector system

Self inactivating (SIN) LTR

Non harmful insert

Exclusion of sharps

Low viral titres

2If at least one of

the criteria applies

the activity will be

Class 2

1st/2nd generation vector system

Vectors containing X-protein expressing forms of the woodchuck

hepatitis B virus post-transcriptional regulatory element

Harmful insert

Use of sharps required*

High viral titres and/or aerosol generating procedures warranting the

use of a microbiological safety cabinet**

*’Whilst ‘control of sharps’ is not one of the specified control measures in the containment tables, other containment level 2 measures are deemed necessary to facilitate their control (e.g. restricted access to authorised and trained personnel; written training records; and the use of gloves) therefore necessitating a classification of Class 2.’ (SACGM compendium of guidance).**Does not apply if the use of the cabinet is solely for purpose of ensuring the sterility of the work.

Peer Review Procedure Occupational Health

& Safety Service

£981!

Peer Review Procedure Occupational Health

& Safety Service

£734!

Changes to:• Host• Vector• Insert• Personnel• Location• Closing date

And finally……… Occupational Health

& Safety Service

Let us know when your GM work has ceased

AND

The samples have been destroyed or transferred elsewhere

We must tell HSE when activity class 2/3 projects have ceasedWe can submit a transfer notification to HSE if you move to another universityWe need to keep the GM register up to date

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Any Questions?