Download Personnel changes should be notified to GMSO

Guidance Notes for GM Risk Assessment
University of Glasgow Garscube Estate
Faculty of Veterinary Medicine : Wellcome Surgical Institute
Risk Assessment For Genetic Modification Under The Genetically Modified
Organisms (Contained Use) Regulations 2000
This form covers work with all GM microorganisms in the laboratory and in animals. A separate
form is available for work with GM animals (e.g. transgenic mice). For projects involving GMOs in
GM animals, please complete a RA considering risks associated with the GMOs (in the lab and
animal facilities) and a GMA RA considering risks posed by the GM animals in their own right.
CLASS OF ACTIVITY
Class 1,2,3 or 4
= maximum containment level assigned to the project through necessity to protect human
health and the environment : containment level 2 requirement = Class 2 project
[NB there will be instances in which level 2 is applied for practical rather than safety reasons
e.g. handling of cells in Class 2 hood, performance of Class 1 activities at level 2 (since most
Vet School labs are at level 2) : in such cases class of activity is not necessarily containment
level applied]
PROJECT TITLE
projects involving use of GMMs in GM in GM animals require submission of 2 risk
assessments, one for GMMs and one for GMAs. If you intend using specific GMAs in a new
project, check with Biological Services staff or GMSO to determine whether a preexisting risk
assessment exists for the animals.
For class 2 and 3 projects consider submitting a generic risk assessment if possible, to save
money ! – e.g. analysis of Canine adenovirus mutants, rather than analysis of an E1 deletion
mutant of canine adenovirus : the HSE may charge upwards of £300 for modifications to
assessments previously notified to HSE.
GMOs
in animals ?
Laboratory bacteria
(i.e. standard commercial disabled e.coli
strains e.g. Top10, JM101)
/DNA vaccine
DNA vaccines are not regarded as GMOs.
However, the bacteria used to generate them
are, so a RA is necessary.
bacteriophage
bacteria ( e.g. genetically modified salmonella
spp)
baculovirus
virus
lower eukaryote
No
Yes
delete non applicable category
If yes :
Quote RodentRisks RA for any work involving
rodents [contact [email protected] for
copy]
..plus quote specific GMA risk assessment if
GMOs to be applied to GMAs
delete non applicable categories
GENETICALLY MODIFIED ORGANISM(S)
e.g. E.coli JM9 +pCR2.1 plasmid containing dog IFN-gamma cDNA
e.g. Canine adenovirus with deletion in E3 region
PERSONNEL
PRINCIPAL APPLICANT
project supervisor or main worker
DEPARTMENT/UNIT
VCS/VPCS/Path/Para/WSI
GROUP/SECTION
if applicable
Additional workers [include Biological Services staff if appropriate]
Names of additional workers will be recorded in a GMSC database – only Principal Applicant
will be named in the archived RA.
Personnel changes should be notified to GMSO [email protected] so that the
personnel database can be updated.
Laboratories where the work is
to be carried out
Containment
Level
Activity
Activity Class
Animal Facilities where the
work is to be carried out *
Containment
Level
Activity
Activity Class
Containment level (and Class) of activity may differ from the actual containment level of the
facility - the latter should always be higher than the former.
* a copy of the finalised risk assessment will be sent to the animal facility by GMSO
Genetic Modification Safety Committee Use Only
DESIGNATION
ACTION
HSE
1. Description of Project
Give details of sources of DNA procedures and vectors
Class 1 projects - 1/2 -1 page of description of project, strain types, vector types, host types etc.
Class 2/3 projects - more detailed description of project and constructs with background
information and referencing as appropriate.
The description should be clear to someone with a biological background outwith the project
discipline.
1. Risk assessment for human health and environmental safety
Hazard group of parental organism – if applicable
see ACDP Categorisation of biological agents according to hazard and categories of
containment for hazard grouping for human pathogens;
also for listings of pathogens controlled by MAFF
Often the containment level can be deduced from the hazard group of the unmodified GMO but consider whether the genetic modification performed might increase (or decrease) the
hazardous nature of the modified GMO.
Brenner Scheme [ NOT applicable to viral vectors ]
See ACGM Guidance part 2A-Annex I,II
Brenner scheme : applicable to GMOs produced by the introduction of a foreign gene(s) into a
laboratory E.coli strain (or yeast/cell line). It is not applicable to insertion of genes into viral
vectors. Provisional containment level is determined by multiplication of Access, Expression
and Damage factors. For work with non-laboratory hosts such as salmonella where the
intended mutations will result in a GMO of unknown pathogenicity this scheme may
underestimate the required containment level. In this case and, in general, it is better to
deduce containment level on the basis of a full assessment of risk to human health and the
environment (i.e. through determination of containment level which reduces Risk of Hazard to
low or effectively zero).
Brenner Scheme Total
CONTAINMENT LEVEL
-15
10
1
-12
10
2
-9
10
3
10-6
3 or 4 case by case contact HSE
>10-6
4
Access
Expression
Damage
Total
This section considers the risk to humans and the environment posed by exposure to the
GMO (risk in context of containment is considered in next section)
Risk to human health presented by host, vector, insert and final GMM
Bacteria
For bacteria : specify disabled nature of host, mobilisation properties of plasmid vector :
assignment of common lab strains of E.coli etc.and vectors is listed in Part 2A Annex II-if strain
is not listed , consult (and cite where appropriate) manufacturer’s information/literature.
Please comment on basis for application of Brenner Scheme values under headings:
Access - likelihood of GMO entering and surviving in a human
Expression – promoter used (if any), expected level of expression of cloned protein in human
Damage - potential for harm to humans [activity of inserted nucleic acid/product]
Viral GMOs : consider tropism of GMM and disease causing potential in animals and man : if
retrovirus vectors + packaging line, risk of replication competent virus generation [defective
retroviruses produced from packaging cell line in which the helper genes are located in two
separate blocks of DNA are regarded as inherently safe GMOs by HSE – Class 1 – but
consider effect of foreign genes inserted into such vectors - insertion of oncogenes increases
Class to 2]. Viruses generated by transfection of cloned genomic/subgenomic fragments are
regarded as GMMs even if no deliberate manipulations such as deletions of genes/insertion of
foreign DNA have been performed. Viruses generated by transfection of DNA purified from
virion preparations (uncloned) are not regarded as GMM.
Full assessment - points for consideration : host
 does GMO infect humans, is it listed in ACDP hazard group or a pathogen controlled by
MAFF?
 pathogenicity of host strain
 presence of adventitious agents (cell cultures)
 for human pathogens good starting point is hazard group value - justify lower containment
level if pathogenicity is known to be reduced in GMO relative to parental organism
(reference literature)
 any possibility of reversion/complementation ?
 does the nature of the work preclude it being undertaken by any workers who have a
serious skin condition or immunological deficiency ?
points for consideration : foreign gene
 does insertion of foreign gene alter pathogenicity of host through complementation/
transcriptional effects ?
 does the inserted DNA encode a toxin, oncogenic protein, allergen, hormone, cytokine,
antibiotic resistance factor or any other protein with a potentially harmful biological activity ?
 is it possible that expression of the inserted gene could alter tissue tropism, host range or
infectivity? [if not expressed/expressed in inactive form (e.g. human gene product of which
requires post translational modifications not performed in bacterial expression system)
unlikely that it will cause harm]
 for non-standard procedures such as animal inoculation consider exposure to risks such as
needle stick injury, in vivo recombination/reversion events (if applicable)
Comment on consequence of human exposure to GMO [See Table below] – severe, medium,
low or negligible ?
Assignment of provisional containment level
This is the initial containment level assigned based on the risk to human health only –
generally this will be based on the hazard group of the pathogen taking into account whether
any changes increase/decrease risk to human health.(e.g. insertion of oncogenes into viral
vector may increase containment level necessary by 1 increment; rendering group 2 pathogen
replication defective may decrease containment level necessary by 1 increment).
Animal pathogens with no risk to human health would be assigned 1 at this stage but as a
general rule they would be reassigned minimum Class 2 after assessment of environmental
risk.
Risk to environment presented by host, vector, insert and final GMM
Risk assessment has to take into account an assessment of the degree of potential harm and severity
of consequence, and the likelihood, or frequency, of that harm occurring. This section considers the
first part, Section 2 considers the latter part.
Assume worst case scenario – if GMM escaped into environment what could happen ? (control
measures employed to minimise this risk are detailed in section 2)
points for consideration :
routes of escape of GMM - waste disposal routes
 probability of harm to the environment (land/sea/air) arising as a result of escape of
GMO from containment facility
 could the GMO survive in the environment in the event of a breach of laboratory
containment ?
 consider animal species infected by GMO and its pathogenicity within these species
 potential for genetic transfer between GMM and organisms in environment
 products of gene expression
 stability (phenotypic and genotypic)
 capacity of GMM to establish, disseminate and/or displace other organisms

numbers of organisms being handled
 adverse effects arising from natural transfer of inserted genetic material toother
organisms
In many cases, the characteristics of the host organism will be more relevant to a risk assessment than
those of the donor organism (e.g. cloning of T. Rex DNA into pCR2.1 and transformation of E.coli). As
a general guide, if a donor organism is merely used as a source of well characterised DNA …. the
characteristics of the donor will not need to be considered. If, however, the insert contains genes
encoding biologically active molecules, toxins or virulence factors, then relevant information from the
donor organism should be considered.
Comment on consequence of environmental exposure to GMO [See Table 1] – severe,
medium, low or negligible ?
This section details the containment level which must be applied to reduce the risk to
human health and the environment (which includes animal health) to an acceptible level
(i.e. low or effectively zero using HSE criteria: see Table 1)
2. Assignment of containment level
Maximum culture volumes
State maximum culture volumes/organism numbers/ virus titre
Control measures undertaken to protect human health and environment
Activity
Control measures
example :
infection of cell culture with virus
performed in Class 2 hood to prevent escape
into environment and exposure of operator to
aerosols
Disposal and spill procedures
The new regulations place increased emphasis on the inactivation of GMOs prior to disposal
and it is important that the RA details inactivation protocols for all GMOs in the assessment
e.g. disposal of GMO waste (liquid and contaminated plastic/glassware) [some information
regarding disposal of Class 1 bacteria is on the local GM web site]
disposal of animal carcasses and waste (urine/faeces/bedding etc.), where contamination with
GMOs is applicable
Provide information for laboratory and animal facilities (ACGM Guidance part 3E)) :
Define inactivation methods to be applied and spill control (state ppm or refer to local code of
practice if standard levels are suitable for inactivation of the GMO). Evidence for efficacy of
disinfectant should be presented (refer to manufacturer’s data or your own data). Autoclaving
is assumed to be effective for most bacterial and viral GMMs.
For Class 2 projects covered by the codes of practice for the Vaccinia laboratory, LRF
containment lab or Cat 3 lab, refer to these codes of practice for general procedures but
specify disposal and spill protocols.
Other issues : transportation of GMOs/infected animals within departments/across campus.....
Assignment of final containment level
this section considers if there are any reasons for applying a higher containment level (CL)
than the provisional containment level defined above to protect the environment – e.g.
activities at large scale may increase required CL, MAFF-controlled pathogens may need
higher levels of containment to minimise risk of environmental release. Animal pathogens are
regarded as minimum Class 2 unless justification can be made for genetic background which
renders them less hazardous to animals (i.e. ‘the environment’).
Cite containment level of animal facilities if GMOs being administered to animals
See Schedule 8 for info on containment levels : note that most labs and working practices in
the Vet School are currently classified as Containment Level 2 [WSI and Vet Path are CL2 with
facilities such as Vaccinia laboratory and cat3 lab in Vet Path providing additional safety
measures with respect to working practices, disposal of GMOs and access to the facility].
Where Level 2 is applied for the sake of convenience rather than safety, and work involves
GMOs which could be safely handled at Level 1, then Class 1 is the appropriate activity class.
If some measures to be applied at lower containment level, state measures and justify
Projects must be classified according to the HIGHEST level of containment applied for reasons
of safety. However, some projects may involve a number of procedures which can be safely
carried out at a lower level - such procedures should be detailed and the use of lower
containment level justified
Does application of containment measures reduce risk of hazard to low or effectively zero ?
If your initial assessment of risk is not 'low' or 'effectively zero', containment level should be
reviewed and risk assessment revised accordingly.
Table 1
Consequence
Severe
Medium
Low
Negligible
High
High
High
Medium/Low
Effectively zero
Likelihood of
Medium
High
Medium
Low
Effectively zero
Hazard
Low
Medium
Medium/Low
Low
Effectively zero
Negligible
Effectively zero
Effectively zero
Effectively zero
Effectively zero
This part of the risk assessment is provided for your convenience – it offers a simple way of
recording that personnel involved in the work have read the RA and GM Local Code of Practice.
Its use is optional and you may choose to record training of staff in another manner.
4. Training Record
All staff involved in the work should read this Risk Assessment and the GM Local Code of
Practice and sign below to indicate they are aware of the hazards associated with this project
and the measures necessary to minimise risk to human health and the environment posed by
the GMO(s). The RA should be retained by the Principal Investigator/Safety Officer/Lab
Manager so that staff can access the RA for reference purposes.
Name
Signature
Date