ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Amended under s67A on 23 August 2007
Application Code
GMD99002
Date
21 July 2000
Consideration Date
11 May 2000
Considered by
Genetically Modified Organism (GMO) Standing Committee of
the Authority
Application Details
Applicant
Application Category
Purpose
Date Application Received
University of Otago
Development in Containment any Genetically Modified Organism
To develop in containment replication-deficient recombinant retroviruses
containing a range of DNA. The retroviruses will be inserted into cell
lines to study the role of viral and cellular proteins in cell cycle control.
1 April 1999
Decision
The application is Approved with Controls.
The organisms approved are:
1.
Escherichia coli strains K12 and B and their derivatives that do not contain conjugative
plasmids or generalised transducing phages, as modified by cloning vectorsa as listed in
Annex 11 and containing nucleic acids sourced from the following items:
i.
Papillomavirus genes E1, E2, E4, E5, E6, E7, L1, and L2.
ii.
Adenovirus genes E1, E2, E3, E4
iii. The ribosome entry site sequence expression cassette from pIRES1neo
iv. Genes that are involved with cell cycle controlb, sourced from eukaryotesc
and being organisms the development of which shall meet Category A or B experiments as
defined in the Hazardous Substances and New Organisms Act (Low-Risk Genetic
Modification) Regulations 1998.
1
Annex 1 refers to the Appendix 1 of the application GMD99002
2.
Replication defective but infection-competent retroviral (Retroviridae) vectors as modified
by:
i.
(a)
(b)
Papillomavirus genes E1, E2, E4, E5, E6, E7, L1, and L2, or
Adenovirus genes E1, E2, E3, E4
ii.
The ribosome entry site sequence expression cassette from pIRES1neo
iii. Genes that are involved with cell cycle controlb, sourced from eukaryotesc
Under no circumstances shall two known oncogenes be included within the same vector.
3.
Epitheloid and fibroblast cells from mouse (Mus musculus), rat (Rattus norvegicus), and humand
as modified by the retroviral vectors identified in 2 above.
4.
Epitheloid and fibroblast cells from mouse (Mus musculus), rat (Rattus norvegicus), and human
as modified by E1b deficient adenoviruses and the retroviral vectors identified in 2 above.
5.
Mouse and humand retrovirus packaging cell lines.
a
b
c
d
The approved vectors shall not include plasmids that are able to transfer themselves by conjugation or
generalised transducing phages.
Genes involved in cell cycle control are those that play a role in the regulation of cellular growth or that
are toxic to mammalian cells.
Genetic material originally sourced from Māori and native or valued flora and fauna shall not be used.
Genetic material from CITES-listed species shall not be used without the written permission of the
Department of Conservation.
Note that the development of genetically modified human cell lines does not require an approval under
the HSNO Act 1996. The definition of organism in the HSNO Act, explicitly, does not include a human being or
a genetic structure derived from a human being.
Application Process
The application was formally received on 1 April 1999. The application was stalled on 16 April
1999 whilst additional information was sought from the applicant. The application was verified
on 31 March 2000.
The application was not publicly notified.
The documents available for the evaluation and review of the application by ERMA New
Zealand included; the application, appendices (including copies of all literature cited), general
reference material on retroviruses, material from other jurisdictions including the Genetic
Manipulation Advisory Committee (GMAC) Guidelines for Small Scale Genetic Manipulation Work
April 1995 (Australia), and comments from the Department of Conservation (DoC).
The application was determined by the Genetically Modified Organisms Standing Committee of
the Authority appointed in accordance with section 19(2)(b) of the Hazardous Substances and
New Organisms (HSNO) Act 1996. The Committee comprised members of the Authority:
Professor Barry Scott (Chair), Helen Hughes, and Dr Oliver Sutherland.
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Additional Information
In the course of its consideration the Committee agreed that further information was required on
aspects of the application. Information was requested primarily with respect to the occupational
risk to personnel working with these organisms, and specifically on the adequacy of the proposed
containment controls to minimise this risk.
It was agreed that an expert virologist review and comment on the following aspects of the
application:

the adequacy of the proposed controls to manage the risks associated with the organisms,
principally the occupational risk to workers

any additional risks associated with the applications not identified in the ERMA New
Zealand Evaluation and Review, and any additional measures required to manage those
risks
In addition, advice was sought on the appropriateness and feasibility of requiring the applicant to
test for the production of replication-competent retroviruses in cells and in the packaging cell
lines to be utilised in the development of genetically modified retroviruses.
Information received from the external reviewer on the adequacy of the proposed controls was
considered by the Committee.
No additional risks were identified by the reviewer that had not been identified in the ERMA
New Zealand Evaluation and Review Report.
Relevant Legislative Criteria
The application was lodged pursuant to section 40(1)(b) of the HSNO Act 1996.
The matters considered in making this decision comprise those set out in sections 37, 44 and 45 of
the Hazardous Substances and New Organisms Act 1996, and those relevant matters in Part II of
the Act.
Consideration of the application followed the relevant provisions of the Hazardous Substances and
New Organisms (Methodology) Order 1998.
The Application
Scope of the application
The application seeks approval for a multi-stage programme; including the development of
genetically modified E. coli, retroviruses, and the development of the genetically modified cell
lines.
The development of specified categories of genetically modified organisms, as defined in
Category A and B of the Hazardous Substances and New Organisms (Low-Risk Genetic Modification)
Regulations 1998 are able to be considered by an Institutional Biological Safety Committee (IBSC)
under delegated authority from the Environmental Risk Management Authority (the Authority).
However Category C experiments as defined in the regulations cannot be considered under
delegated authority.
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The development of the genetically modified retroviruses as a part of this application is required
to be considered by the Authority, as the manipulation techniques and materials to be utilised
make the development fall within the definition of a Category C (c) experiment.
A number of the other stages of the programme, including the development of genetically
modified E. coli, are not Category C experiments and could have been considered by an IBSC,
however they have been included in this application in order that the programme as a whole be
considered as.
Identification of the organisms
In the case of this application not all of the genetic modifications are fully described by the
applicant. The range of foreign genes to be introduced into the organisms has not been
completely specified, as this can only be determined as the research proceeds. However, the
applicant has identified the general sources of the genes that will be used, ie papillomaviruses,
adenoviruses and eukaryotes (primarily mammals). In addition, the applicant has identified that
they are concerned with genes that will assist them to study the role of viral and cellular proteins
in cell cycle control.
The Authority accepts that in the case of the development of genetically modified organisms an
exact identification of all of the elements of the organisms may not be able to be specified at the
outset of the research. However this approval (in the description of the organisms approved on
page 1) details the organisms that may be developed and the range of the genetic modifications
that can be introduced.
This position is consistent with policies developed by the Authority for the identification of
organisms. The ERMA New Zealand Protocol Interpretations and explanations of key
concepts (Number 3, Series 2) provides for this type of identification specifying that… if a
complete taxonomic description is difficult or problematic the Authority will, within the latitude provided by the
requirements of the HSNO Act, require a description of the biological properties of the organism. In general, it
will require relatively more detailed information where the potential risks are high and relatively less detailed
information where the potential risks are low.
Adequacy of the Proposed Containment Regime
In considering the adequacy of the proposed containment regime the Committee considered, inter
alia:
i.
the ability of genetically modified organisms (including Escherichia coli strains K12 and B,
genetically modified replication-defective but infection-competent retroviral vectors, and,
cell lines) to escape from containment, including inter alia:
(a)
the risk of inadvertent infection of personnel working with the organisms
(b)
the adequacy of PC1 and PC2 containment levels for containing organisms under
this approval
ii.
the ability of the organisms to establish self-sustaining populations
iii.
the ease of eradication of any populations established.
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Ability of genetically modified organisms to escape from containment,
establishment of self-sustaining populations and ease of eradication
In accordance with section 44(b) the Committee considered the ability of genetically modified
organisms as a part of this application to escape from containment. The principal mechanisms of
escape considered by the Committee included:
i.
inadvertent release, via human error or failure of procedures
ii.
deliberate action (sabotage)
iii.
natural disaster
iv.
infection of personnel working with the organisms.
And furthermore, having escaped that the organisms would fail to be recovered as a result of:

deficiencies in procedures, including identification of material

inadequacy of contingency plans

infection of personnel.
The organisms are required to be contained in an approved containment facility (under the
Biosecurity Act 1993) in compliance with the Ministry of Agriculture and Forestry (MAF)
Biosecurity Authority (Animal Health and Welfare) Standard 154.03.02: Containment Facilities for
Microorganisms. This standard is employed to meet the requirements of the Hazardous Substances
and New Organisms (HSNO) Act 1996. In addition the operation, construction and management
of the facility must be in accordance with the relevant provisions of the Australian/New Zealand
Standard (AS/NZS) 2243.3:1995 Safety in Laboratories Part 3: Microbiology at Physical Containment
Level 1 or 2. Furthermore, the applicant must comply with the additional controls imposed by
the Authority in this decision.
The principal pathway for escape of the organisms from containment, other than infection of
workers, would be inadvertent, or deliberate removal of the organisms from the facility.
A condition of facility registration is that the approval holder (facility Operator) must prepare,
maintain and implement a quality assurance programme and procedures that address the
requirements of the standard, including the operating requirements. The manual and operating
procedures approved by the facility Supervisor2, are required to address the containment
requirements (including how controls imposed by the Authority are to be met), training
requirements for personnel working within the facility, and internal quality systems in place.
In addition, to ensure that personnel working with these organisms are aware of, and understand
all of the requirements of containment, including the provisions of the MAF Biosecurity
Authority/ERMA New Zealand Standard, AS/NZS2243.3: 1995, and the additional controls
imposed in this decision, the applicant is required to prepare a laboratory operations manual
(containment manual) for all persons working with organisms under this approval. This
containment manual is required to be available at all times, and all personnel are required to sign
2
An inspector appointed under the Biosecurity Act.
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a declaration stating that they have read and understood the relevant standards, and the additional
controls specified in this approval.
Taking into account that the applicant has experience in developing and using these organisms in
containment for research purposes, and additional controls imposed to ensure that all personnel
are adequately trained in the containment requirements, the Committee was satisfied that the
likelihood of an escape of these organisms from containment by inadvertent removal is very low.
The Committee was satisfied that the containment regime in place would equally minimise the
likelihood of deliberate action (sabotage) or natural disaster leading to a breach in containment.
Inadvertent Infection of Personnel Working with the Organisms
The Committee considered that the principal risk associated with this application is the risk that
personnel working with these organisms could become inadvertently infected with the retroviral
vectors they are handling. Such an infection route would also constitute a pathway for the escape
of the organism from containment.
The vectors included in this application are replication-defective (in that they lack essential gene
products that would enable them to propagate) and therefore are only able to infect cells they
directly come in contact with and not be able to produce more infective particles.
However replication-competent retrovirus could be formed if recombination occurs between the
retroviral vector and an (undetected) integrated retrovirus in the cell(s) used. This recombination
event could restore the deleted genes into the vector and make it a fully functional retrovirus. The
applicant has indicated that it is not possible to guarantee that the cell(s) and cell lines to be used
will be retrovirus-free, so that there exists a prospect of replication-competent viruses being
formed.
However the probability of this occurring is considered to be low. Retroviral vectors have been
used since the early 1980s and published reports of reconstitution of replication competence are
uncommon.
No evidence is available that the cell lines to be used (HeLa and A549) contain any endogenous
retroviruses, and there is no evidence that any replication-competent retroviral vectors have been
generated in either of these cell lines. These cell lines have been used for many years by many
researchers for a variety of retroviral infection (and other) experiments so there is reasonable
certainty that they are safe for use.
Furthermore, considerable research involving retroviral vectors is undertaken internationally and
no major incidents, including infection of workers, have been identified in the scientific literature
available.
In the unlikely event of the generation of a replication-competent retrovirus, and in the event of
that organism escaping from containment, it would be unlikely that the organism would be able
to be recovered or eradicated, and there is a remote possibility that it could spread if the infection
path could not be controlled.
However, the Committee were satisfied that the likelihood of the generation of a replicationcompetent retrovirus is very low.
The Committee noted the comments of the expert reviewer that although it was uncommon for
a recombination event between a recombinant replication-deficient retrovirus and an endogenous
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retrovirus in a packaging system to occur, that personnel working with these organisms should be
adopting containment measures on the assumption that the stocks and cells do potentially
contain replication-competent retroviruses.
All experiments involving work with retroviruses will be conducted under PC2 containment
conditions, as specified in the AS/NZS 2243.3: 1995. The application states that the experiments
will also be undertaken in accordance with the Australian Genetic Manipulation Advisory
Committee Guidance for work involving genetically modified viruses for gene transfer into animal and human
cells (Appendix 5 of the Guidelines for small scale genetic manipulation work). This document provides
guidelines based on good virological and tissue culture techniques. The Committee has reviewed
the guidelines and incorporated a number of the elements into the controls to which this
approval is subject.
These additional control measures over and above the requirements of PC2 containment are
designed to further minimise the possibility of infection of personnel working with retroviruses,
and include:

the use of a biological safety cabinet of class II (refer AS 2252.2 3) for all experiments
involving the handling of retroviruses

handling of dishes and plates of cells containing human-infectious viruses in larger plates
(or inverted lids) to provide traps for accidental spills

storage of tissue cultures infected with human-infectious or potentially human-infectious
viruses in incubators dedicated to the use of human-infectious viruses

storage of human-infectious viruses or infected cell lines in a section of the freezer
specifically designated for this purpose and clearly marked to this effect
Furthermore, additional controls have been imposed in this approval requiring the facility
Operator in conjunction with the IBSC, to ensure that only suitably qualified trained personnel
may handle human-infectious recombinant viruses, and that their qualifications and training
records be available for inspection.
Taking into account the additional controls imposed in this decision derived in part from the
GMAC Guidelines, and the specific requirements with respect to the training and qualifications
of personnel working with genetically modified retroviruses, the Committee considered that
probability of a worker being infected is low. The key factor in enabling this risk to be managed
to a low level, however, is the management and oversight by those responsible for ensuring the
controls imposed in this decision are rigorously adhered to.
Adequacy of PC1 and PC2 containment levels for containing genetically modified
organisms under this approval
As noted above this application covers the development of a range of genetically modified
organisms, including E. coli, retroviruses, and cell lines that each present differing risk profiles.
3
AS 2252.2 Part 2: Laminar flow biological safety cabinets (Class II) for personnel and product protection
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E. coli
The low-risk regulations referred to above, the Hazardous Substances and New Organisms (Low-Risk
Genetic Modification) Regulations 1998, specify the appropriate level of containment for the
development of genetically modified organisms in accordance with the manipulation techniques
and materials utilised. Theses regulations state that where certain techniques are performed
(defined as Category A or B) using host/vector systems listed in Schedule 2 of the regulation the
appropriate level of containment is Physical Containment Level 1 (PC1) as defined in the in
2243.3:1995.
Schedule 2 of the HSNO Low-Risk Regulations (referred to above), Approved Host/Vector Systems,
identifies as an approved host E. coli K12 or E. coli B derivatives that do not contain conjugative
plasmids4 or generalised transducing phages5. Approved vectors for use with these E. coli strains
are either non-conjugative plasmids or specific bacteriophage6. The particular nucleic acids used
in each modification will determine whether Category A or B applies, however PC1 is the
required containment for both, where approved host/vector systems are used.
Controls to which this approval is subject therefore require that the development of genetically
modified E. coli be undertaken at Physical Containment Level 1 (PC1) as defined in the
Australian/New Zealand Standard 2243.3:1995.
Retroviruses
The development of the retroviruses, and infection of the cells lines constitute Category C
experiments and are required to be undertaken at Physical Containment Level 2 (PC2) as defined
in AS/NZS 2243.3:1995.
In addition, as a result of the risk of infection of personnel working with these organisms the
Committee has imposed a range of additional controls to address particular issues of training and
the qualifications personnel working with these organisms, and other measures to minimise the
prospect of inadvertent infection occurring.
The Committee was satisfied that adherence with the requirements of the relevant standards
including the MAF Biosecurity Authority/ERMA New Zealand containment standard, the
relevant sections of AS/NZS 2243.3: 1995, and the additional controls specified, can adequately
contain organisms specified under this approval, and in particular, will minimise the prospect of
an escape from containment via infection of personnel.
Population establishment and ease of eradication
The majority of the organisms covered under this approval are not in themselves able to enter
the natural environment and all microorganisms covered under this approval are required to be
maintained in specialised vessels (petri dishes or tubes) within the containment facility, containing
specific culture media required for the survival of the organism. Retroviruses covered under this
approval cannot survive independently outside of a host cell.
Conjugative plasmid: a plasmid containing a set of transfer genes that promotes bacterial conjugation (transfer of
DNA from one bacteria to another).
4
5
Transducing phages: virus particle containing host cell DNA instead of the viruses’ own genes.
6
Bacteriophage: a virus which infects bacteria and is usually species-specific.
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E. coli
The ability of E. coli to establish a self-sustaining population was considered in the context of
application NOC99007 (to import into containment genetically modified E. coli), approved by the
Authority in March 2000. In the decision on that application it was noted that E. coli strain K12
has been subcultured in research laboratories since 1922 and the closely related strain B has been
subcultured for over 30 years. They are not known to colonise the alimentary tract of a human
host or cause any symptoms of disease. The strains are genetically crippled and therefore cannot
survive without certain growth factors. As the laboratory-based strains of E. coli will be
maintained in approved containment, are unable to survive outside of controlled conditions, and
do not contain self-transmissible vectors, the Committee in that case considered that escape and
establishment of self-sustaining populations, which might cause adverse effects, was extremely
unlikely.
Retroviruses
Replication-deficient retroviruses would be unable to establish a self-sustaining population, as
they are unable to propagate themselves.
For a replication-deficient retrovirus to establish a self-sustaining population it would first be
required to recover its original gene complement, and therefore become ‘replication-competent’.
Retroviruses have very small genomes (only 7-10 kilobases) so that carrying additional foreign
genes will confer a packaging disadvantage on the virus and, consequently, is likely to make it less
infective. An escaped and infective retrovirus is unlikely to carry foreign genes and so any
infected individuals will be at risk primarily from the symptoms caused by that virus.
Any affected individuals may be able to be identified and treated to ameliorate the effects, as well
as controlling the retroviruses spread. Retroviruses are transmitted primarily by direct contact
between infected body fluids and other suitable cells, so that spread of the virus would be able to
be managed.
Cell Lines
The ability of cell lines containing foreign DNA to establish a self-sustaining population was
considered in the context of application NOC99004 (to import into containment genetically modified
mink epithelial cell line modified by human and firefly DNA), approved by the Authority in October
1999. In the decision on that application it was noted that the cell line could not survive
unassisted in the uncontrolled environment. The Committee took into account that the cells
could only survive (i) in a laboratory cell culture system that provides a complex and accurately
balanced mixture of highly purified nutrients and a stable temperature, or (ii) after being placed
by surgery or injection inside an immune system-weakened laboratory animal.
Furthermore, the Committee noted that there has been no recorded instance of a cultured cell
acquiring an ability to naturally infect an animal or plant. It was, however, noted that in the case
of NOC99004 no viral or other infectious agent was used in the genetic transformation of the
cell line. The issue of infection of workers from viral components is addressed in the present
decision.
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Effects of the Organism
As a result of the low likelihood of organisms under this approval surviving or persisting in the
environment, and the low likelihood of the escape of organisms from containment, the principal
effect considered by the Committee was the effect on personnel working with genetically
modified retroviruses in the event of infection.
Occupational risk to workers
This approval covers the use of several different retroviral vectors, some of which are capable of
infecting only a specific host (such as mouse), while others have the potential to affect several
species, including humans.
As the retroviral vectors used in this application are replication-defective, any escape is expected
to result only in the infection of cells that the virus comes in direct contact with. However, as
noted previously, there is a possibility for a replication-competent virus to be created, but it too
requires direct contact with uninfected cells to escape.
Infection of a researcher, or other person, with one of the retroviral vectors (whether or not the
vector is replication-competent) could have one or more of the following effects:
i.
alteration of cellular processes as a consequence of retroviral infection
ii.
expression of the foreign gene in the recipient
iii.
random insertion of the virus into the genome could induce a mutational change (such as
knocking out a gene) in an endogenous gene if the insertion occurred within it
iv.
random insertion of the retrovirus could induce cellular transformation (ie tumour
formation) rather than a simple mutational change.
The most significant of these effects is the possibility of expression of the genes that the vector
contains. The consequences will be larger for a replication-competent vector since this would be
able to spread to other cells.
Most of the foreign genes to be utilised in these experiments are those involved in cell cycle
control, and therefore there is a high likelihood that infected cells would have altered growth
and/or development. The extent to which growth and development of cells would be altered can
not be precisely defined at this point, although considering the nature of the genes involved,
some of the effects on the infected cells may be significant. The nature of the effects would also
be dependent upon the types of cells infected, for example whether they are short-lived epithelial
cells or pluripotent stem cells. Retroviruses are only able to infect cells that have the appropriate
cell surface proteins, so that not all cells will have the same susceptibility to infection. If the
vector is unable to replicate effectively the effect may be limited to the infected cells or tissues.
The effects of such an infection could range from:
i.
non-demonstrable effects, due to the foreign genes having very little effect on the infected
cell(s)
ii.
local effects on the growth or development of the infected cell(s) or tissue that may be
treatable
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iii.
more severe effects such as tumour development or organ function disruption. However,
oncogenesis is considered to be of lower likelihood relative to the other effects because it
often requires separate mutations in two or more genes.
The likelihood of the occurrence of other effects listed aboved is considered to be very much
lower as a result of:

The fact that only about approximately 10% of the human genome appears to code for
specific genes, and therefore there is a high probability that a retrovirus will insert at a
location that will not disrupt genome function.

The fact that the effects of other viruses that commonly infect humans and animals appear
to be largely due to the genes encoded by the virus, rather than mutations introduced by
integration of the virus into the genome.

Viral infections are often effectively controlled by the body’s immune system. In most
individuals virus-encoded proteins will be expressed on the surface of infected cells and so
be targets for destruction by the immune system. However newborn animals or individuals
that are immuno-compromised appear to be most susceptible to developing tumorous
responses to viral infection.
As the retroviral vectors specified in this approval are replication-deficient, they are unable to
propagate in cells (or humans). However, as noted above, there is a possibility the replicationcompetent retroviruses could be produced. The major effects of this situation would be that
more cells could become infected and that the virus is transmitted to other individuals.
Taking into account the advice received from an expert virologist, the Committee considers that
controls and protocols specified in this decision will minimise the probability of infection of
personnel.
Other human health effects
As a result of the low likelihood of the generation of replication-competent retroviruses, and the
low likelihood of the survival of these organisms outside of the specific mediums in which they
are maintained in the containment facility, the risks to human health of other than experimenters
working with the organisms is considered to be negligible.
Inseparable organisms
In accordance with section 45(a)(ii) the Committee considered the effects of any inseparable
organisms.
In this case the issue of inseparable organisms includes any endogenous retroviruses present in
cell lines into which DNA will be transferred by recombinant replication-deficient retroviral
vectors.
The cell lines to be utilised in these experiments are not certified as retrovirus free, and it is not
considered to be feasible to test for the presence or absence of unknown retroviruses.
In reviewing this application an expert virologist noted that it was uncommon for a
recombination event between a recombinant replication-deficient retrovirus and an endogenous
retrovirus in a packaging system to occur. And furthermore, that in the case of in vitro work, the
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most significant effect of such an event would be from the point of view of the interpretation of
experimental results, rather than from the perspective of a risk to human health.
These comments were made in the context of comments noted above that personnel working
with these organisms should be adopting appropriate containment measures on the assumption
that the stocks and cells do potentially contain replication-competent retroviruses.
Taking into account the additional controls imposed in this decision, the Committee was satisfied
that occupational risks associated with the possibility of the development of replicationcompetent retroviruses as a result of the presence of endogenous retroviruses is low.
However the Committee notes that the applicant should take heed of the comments of the
reviewer with respect to assuming that replication-competent retroviruses may be present, and
take any additional measures over and above the requirements of this approval in order to protect
personnel from the risk of infection.
Negligible risk
Infection of personnel working with genetically modified retroviruses could result in adverse
health effects, and although the likelihood of such an infection occurring is low, the Committee
concluded that the occupational risk to workers could not be considered negligible.
In evaluating this risk to workers the Committee considered the characteristics of this risk, noting
that exposure to the risk was predominantly voluntary, any adverse effects to the health of the
worker would likely not persist, any adverse effects would be unlikely to spread, any adverse
effects would likely be treatable, and the risk is known and relatively well understood by those
working with the organisms.
The Committee also concluded that, taking into account the containment regime and additional
controls imposed in this decision, the risks to the environment and wider public health associated
with the development of these organisms, are negligible provided careful and strict management
is maintained.
Benefits
The principal direct benefit to be derived from the development in containment of these
organisms was identified as being the generation of scientific knowledge that may answer
questions about how viruses cause cells to die. The value of this information was considered to
be potentially significant in that it may lead to a better understanding of the normal cellular
controls of growth and death.
The Committee acknowledges that the degree of benefit to be derived from this research, as with
all fundamental research, is difficult to quantify. However, that is not to say that the knowledge
accumulated in the research is not beneficial as that information adds to the pool of knowledge
from which other benefits flow, and the Committee accepts that exploratory research is an
essential prerequisite for scientific progress.
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In addition the Committee took into account other indirect benefits that may accrue from the
work to be undertaken under this approval, including:

possible future applications in therapeutic strategies in the treatment of disease such as
cancer

enabling New Zealand scientists to undertake innovative research projects

maintaining New Zealand researchers’ involvement in the international scientific
community maintaining New Zealand’s standing in international science.
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Conclusion
Pursuant to section 45(1)(a)(i) of the Act, the Committee is satisfied that this application is for one
of the purposes specified in section 39(1) of the Act, being section 39(1)(a), The development of any
genetically modified organism.
In terms of clause 27 of the Methodology where risks are deemed not negligible, the Authority
must take into account the extent to which the risks and any costs associated with the organism
may be outweighed by benefits. And, in accordance with section 45(1)(a) may approve the
application if the beneficial effects of having the organism in containment outweigh the adverse
effects of the organism and any inseparable organism should the organism escape, taking account
of the ability of the organism to escape from containment.
In this case the Committee formed the view that the occupational risk to workers associated with
the development of genetically modified retroviruses is not negligible. However, taking into
account the characteristics of the risk, principally that the risk is voluntary and understood by
personnel working with the organisms, and the low likelihood of any adverse effect occurring, the
Committee considered that the risk to workers to be low, provided careful management is
maintained.
The Committee concluded that risks to the environment and wider public health from the
development of these organisms in containment are negligible.
In the absence of any costs to other parties, the Committee weighed the benefits of the proposal
against occupational risk to workers, and concluded that the benefits, principally the generation
of scientific knowledge to be gained outweigh the low risk to workers, taking into account the
voluntary nature of the risk, and the additional risk management measures imposed in controls to
which this approval is subject.
The Committee is satisfied that the proposed containment regime, including compliance with
MAF Biosecurity Authority Standard 154.03.02 and the relevant sections of AS/NZS 2243.3:
1995 Safety in Laboratories. Part 3: Microbiology, and the additional control conditions imposed in
this decision will adequately contain the organism.
Environmental Risk Management Authority Decision: Application GMD99002
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Controls
In order to provide for the matters detailed in Part I of the Third Schedule to the Act, Containment
Controls for Development and Field Testing of Genetically Modified Organisms, this application is approved
subject to the following controls:
1. To limit the likelihood of any accidental release of any organism or any
viable genetic material7:
For the development of genetically modified E. coli
1.1
The construction, operation and management of the containment facility shall be in
accordance with the:
(a)
Ministry of Agriculture and Forestry (MAF) Biosecurity Authority/ERMA New
Zealand Standard 154.03.028: Containment Facilities for Microorganisms.
(b)
Australian/New Zealand Standard (AS/NZS) 2243.3:19958 Safety in Laboratories: Part
3: Microbiology, at Physical Containment Level 1 (PC1).
For the development of genetically modified Retroviridae and genetically
modified cell lines
1.2
The construction, operation and management of the containment facility shall be in
accordance with the:
(a)
MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.028: Containment
Facilities for Microorganisms.
(b)
Australian/New Zealand Standard (AS/NZS) 2243.3:19958 Safety in Laboratories: Part
3: Microbiology, at Physical Containment Level 2 (PC2).
1.3
The facilities shall be approved by MAF as containment facilities in accordance with the
MAF/ERMA New Zealand Standard 154.03.028, and the controls of the Authority, prior
to the development of any genetically modified organisms that are the subject of this
approval.
1.4
A biological safety cabinet of class II (refer AS 2252.29) shall be used for all experiments
requiring PC2 containment involving the handling of retroviruses.
1.5
Dishes and plates of cells containing human-infectious viruses shall be handled in larger
plates (or inverted lids) to provide traps for accidental spills.
Viable Genetic Material is biological material that can be resuscitated to grow into tissues or organisms. It can be defined to
mean biological material capable of growth even though resuscitation procedures may be required, eg when organisms or parts
thereof are sublethally damaged by being frozen, dried, heated, or affected by chemical.
7
Any reference to this standard in these controls refers to any subsequent version approved or endorsed by ERMA
New Zealand.
8
9
AS 2252.2 Part 2: Laminar flow biological safety cabinets (Class II) for personnel and product protection
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1.6
All pipettes, glassware, plasticware, and other used equipment shall be decontaminated by
submersion in a suitable disinfectant or by placing in polyethylene bags that should
subsequently be sealed and autoclaved. Sharps (eg needles and scalpel blades), shall be
disposed of in puncture-resistant containers (refer AS 403110) and incinerated.
1.7
Tissue cultures infected with human-infectious or potentially human-infectious viruses shall
be kept in incubators dedicated to the use of human-infectious viruses.
1.8
Human-infectious viruses or infected cell lines shall be stored in a section of the freezer
specifically designated for this purpose and clearly marked to this effect. Similarly,
ampoules of frozen infected cell lines shall be stored in a separate section of the liquid
nitrogen tank.
2. To exclude unauthorised people from the facility:
2.1
The applicants shall comply with the requirements contained in the standards listed in
controls 1.1 and 1.2 relating to identification of entrances, numbers of and access to
entrances, and security requirements for the entrances and the facility.
3. To exclude other organisms from the facility and to control undesirable and
unwanted organisms within the facility:
3.1
The applicants shall comply with the requirements contained in the standards listed in
control 1.1 and 1.2 relating to exclusion of other organisms from the facility and the
control of undesirable and unwanted organisms within the facility.
4. To prevent unintended release of the organism by experimenters working
with the organism:
4.1
The applicants shall comply with the requirements contained in the standards listed in
control 1.1 and 1.2 relating to the prevention of unintended release of the organisms by
experimenters working with the organisms.
4.2
Under no circumstances should investigators be infecting cultures of their own cells, or of
their immediate relatives, or those of other members of the laboratory.
5. To control the effects of any accidental release or escape of an organism:
5.1
The applicants shall comply with the requirements of the standards listed in control 1.1 and
1.2 relating to the provision of an eradication plan to deal with an accidental release beyond
the facility and into the uncontrolled environment or spillage of microorganisms within the
facility.
5.2
If a breach of containment occurs, the facility operator must ensure that the MAF
Inspector responsible for supervision of the facility has received notification of the breach
within 24 hours.
10
AS 4031 Non-reusable containers for the collection of sharp medical items in health care areas
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6. Inspection and monitoring requirements for containment facilities:
6.1
The inspection and monitoring requirements for containment facilities shall be in
compliance with the standards listed in controls 1.1 and 1.2.
7. Qualifications required of the persons responsible for implementing those
controls:
7.1
The applicant shall comply with the requirements of the standards listed in control 1.1 and
1.2 relating to the training of personnel working in the facility.
7.2
The facility Operator, in consultation with the Institutional Biological Safety Committee
(IBSC), shall ensure that only suitably trained individuals will handle human-infectious
recombinant viruses covered under this approval.
7.3
The facility Operator shall record the qualifications and training undertaken of all
personnel working with organisms under this approval, and make these records available
for examination by the Inspector.
7.4
The Operator shall ensure that every individual working with organisms under this
approval:
i.
has read and understood the requirements of standards identified in controls 1.1 and
1.2, and the additional controls specified in this approval, and in particular those
provisions relating to the safety of personnel working with human-infectious
recombinant viruses.
In order to achieve this, the applicant shall prepare a laboratory operations manual
(containment manual) for all persons working with organisms under this approval,
which is derived from AS/NZS 2243.38 at PC1and PC2, and the additional controls
identified as a part of this approval.
This manual shall be available to all personnel working in the containment facility at all
times.
ii. has signed a declaration that states that they have read and understood the standards
identified in controls 1.1 and 1.2, and the additional controls specified in this approval.
____________________________
D Barry Scott
Chair, GMO Standing Committee
Date: 21 July 2000
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Amendment: November 2006
Changes to controls:
 Addition of footnotes to the containment facility references and the Australian/New
Zealand containment facility references to “future proof” the decision
 Standardise the wording of the breach of containment control
 Replacement of the control regarding inspection of facilities by the Authority, its agent
or enforcement officers with the standard control
____________________________
Dr Kieran Elborough
Chair, GMO Standing Committee
Date: 23 August 2007
Environmental Risk Management Authority Decision: Application GMD99002
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