ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Date signed: 2 October 2007
Application code:
GMD06058
Application category:
Develop in Containment any New Organism under the
Hazardous Substances and New Organisms (HSNO) Act
1996
Institute of Environmental Science and Research
Applicant:
Purpose:
The creation of influenza viruses by reverse genetics in
laboratory containment, for future application in
diagnostics, research and vaccine manufacture
Date received:
21 August 2007
Consideration date:
28 Septmeber 2007
Considered by:
A Committee of the Environmental Risk Management
Authority
1
Summary of decision
1.1
Application GMD06058 to develop in containment, the genetically modified
organisms described in Table 1 is approved with controls (specified in Appendix 1
of this decision), having been considered in accordance with the relevant provisions of
the Hazardous Substances and New Organisms (HSNO) Act 1996 (the Act) and of the
HSNO (Methodology) Order 1998 (the Methodology).
Table 1: Description of organisms approved for development
Host organism
Modified by:
Escherichia coli
(Migula 1895) Castellani and
Chalmers 1919
non-pathogenic laboratory
strains
Standard non-conjugative cloning and/or expression plasmid
vectors containing cDNA sequences encoding the following
the influenza genes:
Nucleoprotein (NP), Matrix protein (M1, and splice variant
M2), Haemagglutinin (HA), Neuraminidase (NA, and splice
variant NB in influenza B viruses), Polymerase genes (PB1,
PB2, PA) and Non-structural protein (NS1, and splice
variant NS2).
Homo sapiens immortalized
cell lines1 (Linnaeus, 1758)
Canis familiaris (dog) cell
lines (Linnaeus, 1758)
Chlorocebus aethiops
(African green monkey) cell
lines (Linnaeus, 1758)
Influenza
(Lamb & Krug, 2001),
Seasonal influenza2 strains
and avian influenza strains3
Vectors may contain one or more of the following: standard
and commercially available promoters, terminators,
regulatory elements, reporter and selectable marker genes,
proteins purification tags and origins of replication.
Plasmids carrying one of the eight influenza gene segments
will be transformed into each E. coli strain.
The mammalian cells will be transiently transfected with
plasmids containing each of the influenza genes.
No vector sequence will be transferred into the Influenza
virus generated.
Alterations to the sequence of the influenza genome may be
necessary for research purposes, to eliminate pathogenic
gene sequences or attenuate the virus. For example: The
pathogenic sequences in the avian HA gene may be removed
through deletion of the polybasic region of amino acids.
These alterations will not increase the pathogenicity,
virulence or infectivity of the host organism.
Reverse genetics to create a virus comprised of two gene
segments (HA and NA and splice variant NB in influenza B
viruses) from pathogenic influenza isolates such as seasonal
human influenza or avian influenza. The further six gene
segments in these influenza strains will be derived from
A/PR/8/34 (or another apathogenic influenza isolate).
Alterations to the sequence of the influenza genome may be
necessary for research purposes, to eliminate pathogenic
gene sequences or attenuate the virus. For example: The
pathogenic sequences in the avian HA gene may be removed
through deletion of the polybasic region of amino acids.
These alterations will not increase the pathogenicity,
virulence or infectivity of the host organism.
Excludes embryonic stem cells or cell lines derived from persons of Māori decent.
Seasonal influenza strains are defined as the influenza A strains that are currently circulating in the human
population as well as all influenza B strains.
3 Avian influenza strains are defined as influenza A viruses that circulate in avian species.
1
2
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2
Legislative criteria for application
2.1
The application was lodged by the Institute of Environmental Science and Research
(ESR) pursuant to section 40(1)(b) of the Act. The decision was made in accordance
with section 45 of the Act taking into account additional matters to be considered
under section 44, and other matters relevant to the purpose of the Act, as specified
under Part II of the Act. Unless otherwise stated, references to section numbers in
this decision refer to sections of the Act.
2.2
The proposed modifications to E. coli and those involving seasonal influenza are low
risk modifications according to the HSNO (Low-Risk Genetic Modification)
Regulations 2003 (the Regulations). However, the application does not qualify for
rapid assessment under section 42A as the modifications to avian influenza involves a
microorganism of Risk Group 3 or above and the production of genomes or fragments
of genomes capable of causing disease in humans, animals, plants, or fungi (other
than those that satisfy the requirements of a category A or B genetic modification).
2.3
Consideration of the application followed the relevant provisions of the Methodology,
as specified in more detail below. Unless otherwise stated, references to clauses in
this decision refer to clauses of the Methodology.
3
Application Process
Application Receipt
3.1
The application was determined to be in compliance with section 40(2) of the Act and
was formally received on 21 August 2007.
Notification
3.2
The Authority has discretion, upon receipt of applications to develop in containment
any new organism, to decide whether or not they are publicly notified (section 53(2)
of the Act). Application GMD06058 was not notified as it was considered that public
notification would be unlikely to generate any information that would be material to
the consideration of this application.
3.3
In accordance with sections 53(4) and 58(1)(c) of the Act and clauses 2(2)(e) and 5 of
the Methodology, the Ministry of Agriculture and Forestry Biosecurity New Zealand
(MAF) were notified and provided with the opportunity to comment on the
applications. MAF raised no issues with this application.
Decision Making Committee
3.4
In accordance with section 19(2)(b) of the Act and clause 43 of the First Schedule to
the Act, the Environmental Risk Management Authority (the Authority) appointed a
committee (“the Committee”) of its members to determine the application. The
Committee comprised of: Dr Kieran Elborough(chair), Dr Max Suckling and Dr
Deborah Read.
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Information available for the consideration
3.5
The information available for the consideration comprised:

Application GMD06058 (Form NO3) submitted by ESR(the applicant);

References as listed in the application;

A memo from the Agency to the Committee to assist and support the
Committee’s decision making; and

Comments received from MAF.
4
Associated Approvals
4.1
The Committee noted that this decision is specific to the applicant and that the
purpose of this application is to create influenza viruses by reverse genetics in
laboratory containment. The Committee noted that for future application of these
viruses, for example in vaccine manufacture, additional HSNO Act approvals would
be required.
5
Sequence of the consideration
5.1
In accordance with clause 24 of the Methodology, the Committee considered the
information provided by the sources listed above. The approach adopted by the
Committee was to look sequentially at identification, assessment and the combined
evaluation of risks and of costs and benefits. Techniques for identifying and assessing
information on risks, costs and benefits were based on internal procedures as specified
in the ERMA New Zealand Technical Guide to identifying assessing and evaluating
risks costs and benefits. Those risks identified as potentially significant were assessed
in accordance with clause 12 of the Methodology. Management techniques were
considered in relation to the assessed risks. Costs and benefits were assessed in
accordance with clause 13 of the Methodology. Qualitative scales used by the
Committee to measure likelihood and magnitude of risks, costs and benefits were
provided in the memo from the Agency to the Committee.
5.2
In carrying out its consideration, the Committee considered the adequacy of
containment in accordance with section 45(1)(a)(iii) of the Act, and the magnitude
and probability of the risks, costs and benefits alongside each other and in an
integrated fashion. This is because the former interacts with the latter and this is
recognized in clause 12(d) of the Methodology and in section 45(1)(a)(ii) of the Act.
5.3
The Committee set controls to satisfactorily provide for the matters in the Third
Schedule (Part I) of the Act (see Appendix 1 of this decision).
5.4
Benefits associated with these applications were considered in accordance with
clauses 9, 10, 13 and 14 of the Methodology and section 6(e) of the Act.
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5.5
Finally, taking account of the risk characteristics established in accordance with
clause 33 of the Methodology, the combined impact of risks, costs and benefits was
evaluated in accordance with clause 34.
6
Purpose of application and scope of the approval
6.1
In accordance with section 40(1)(b) of the Act, the ESR seeks to develop in
containment genetically modified strains of influenza for future application in
diagnostics, research and vaccine manufacture.
6.2
In accordance with section 45(1)(a)(i), the Committee is satisfied that the purpose of
these applications falls within the scope of section 39(1)(a) of the Act: “the
development of any new organism.”
7
Adequacy of the containment regime
7.1
In assessing the ability of the genetically modified organisms (as described in Table
1) to escape from containment, the Committee considered:

The biological characteristics of the organisms;

The genetic modification;

Proposed containment regime; and

Potential pathways for the escape of the organisms from the containment
facility.
Biological characteristics of the host organisms
Non-pathogenic laboratory strains of E. coli
7.2
The non-pathogenic laboratory strains of E. coli that will be used in this research
require growth factors and nutrients for survival and cannot survive outside of
controlled laboratory conditions. The non-pathogenic laboratory strains of E. coli to
be used are not capable of causing disease in humans, animals, plants or fungi, do not
contain infectious agents normally able to cause disease in humans, animals, plants or
fungi, do not normally infect, colonise or establish in humans nor produce
desiccation-resistant structures, such as spores or cysts, and their main biological
characteristics are known.
Mammalian cell lines
7.3
The immortalised mammalian cell lines to be used in this research have highly
specific growth requirements that would limit their ability to escape from
containment. These cells require a controlled atmosphere that is enriched for CO2 and
immersion in cell culture media for survival. Some of the mammalian cell lines may
contain infectious viruses used to immortalise cell lines, such as the adenovirus.
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Seasonal Influenza
7.4
Seasonal influenza strains are defined by the applicant as the influenza A strains that
are currently circulating in the human population as well as all influenza B strains.
Seasonal influenza strains are categorised as Risk Group 2 organisms as per the
Australian/New Zealand Standard: Safety in Laboratories Part 3: Microbiological
aspects and containment facilities (AS/NZS 2243.3:2002) as they can cause human or
animal disease, but are unlikely to be a serious hazard to laboratory workers, the
community, livestock, or the environment. The biological characteristics of influenza
are described in sections 7.6-7.13.
Avian Influenza
7.5
Avian influenza strains are described by the applicant as influenza A viruses that
circulate in avian species. Pathogenic avian influenza strains are considered exotic
animal virus that must be held under quarantine conditions at the CSIRO Australian
Animal Health Laboratory or other facilities approved by AQIS (Australia Quarantine
and Inspection Service) as per the AS/NZS 2243.3:2002. The strains of avian
influenza to be developed in this project will be modified to be apathogenic.
Therefore, they will be unlikely to cause human or animal disease. However,
apathogenic strains of avian influenza do not currently have a Risk Group
classification under the AS/NZS 2243.3:2002. The biological characteristics of
influenza are described in sections 7.6-7.13.
Biological characteristics of influenza
7.6
Influenza viruses are enveloped viruses made up of 8 segments of negative-sense,
single-stranded RNA that encode 10 genes. Two of the RNA segments encode the
envelope glycoproteins, hemagglutinin (HA) and neuraminidase (NA and splice
variant NB for influenza B). HA and NA/NB are important for viral pathogenicity
and are the main antigenic determinants of influenza. Influenza viruses lose viability
quickly (within days) in an ambient environment outside of a susceptible host.
7.7
Negative-sense RNA viruses rely on proteins that are packaged along with the viral
RNA in the infectious virions, to infect a host cell. The proteins required form an
RNA-dependent RNA polymerase that initiates viral replication by transcribing the
negative-sense viral RNAs (which are not infectious) into messenger RNA (mRNA).
The mRNA can then be translated into protein to form new infectious virions. In
reverse genetics the efficiency of virus production can be enhanced by adding protein
expression constructs for RNA polymerase expression. Therefore, by transfecting
cultured mammalian cells with eight plasmids containing viral RNAs and four protein
expression constructs, or with plasmids that can produce both viral RNA and mRNA,
it is possible to produce infectious virus rapidly in cell culture.
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7.8
Influenza viruses evade the immune system through antigenic drift and antigenic shift.
Antigenic drift occurs due to point mutations that result in changes in the amino acid
sequence in the HA and NA/NB glycoproteins. The segmented nature of the genome
also allows for antigenic shift through the formation of reassortant viruses. Rapid
evolution through antigenic shift can occur when one cell is infected with more than
one influenza virus. The viruses can exchange RNA segments and form new
reassortant viruses containing a combination of gene segments derived from the
parental virus strains.
7.9
Influenza viruses infect a wide variety of avian and mammalian species. Host cellular
receptor specificity plays a major part in determining host species restriction. For
example, human influenza strains preferentially bind to sialic acid residues linked to
galactose by an alpha- 2,6 linkage. This is the predominant linkage found in human
respiratory epithelial cells. In contrast, avian influenza strains preferentially bind to
sialic acid residues linked to galactose by an alpha- 2,3 linkage, as is found in avian
host cells.
7.10
Transmission of avian influenza virus between bird species can occur either by direct
contact, or indirectly via faecal-contaminated aerosols, water feed and other materials.
Avian influenza strains may sometimes infect humans who are exposed to large
quantities of virus, even though avian influenza viruses may not efficiently bind
human respiratory cells. If transmission does occur, it is likely to be through
inhalation of infectious droplets, airborne droplet nuclei, or by indirect (fomite)
contact followed by self-inoculation of the upper respiratory tract or conjunctival
mucous membranes.
7.11
Influenza viruses can cause a spectrum of disease ranging from asymptomatic
infection, or mild respiratory illness, through to severe and rapidly fatal systematic
failure. While the factors contributing to the pathogenicity of influenza are not fully
understood, the post-translational cleavage of HA is one important determinant of
pathogenicity. Cleavage is necessary for the spread of infection through the
organism. Non-pathogenic influenza strains typically express HAs that are cleaved in
only a few cell types. Therefore, these viruses only cause local infection. In contrast,
the pathogenic avian influenza strains have polybasic HAs that are cleaved in a broad
range of host cells. Therefore, they can cause systemic infection.
7.12
Influenza virus reassortment in embryonated chicken eggs is currently used in the
manufacture of seasonal human influenza vaccines. Eggs are inoculated with an
influenza virus that grows well in eggs (such as A/PR/8/34) along with the seasonal
influenza strain. However, the production of an appropriate vaccine strain is not
guaranteed and the process requires large arrays of embryonated eggs in order to
produce a strain that grows well and expresses the desired antigens, namely the HA
and NA/NB genes from the seasonal strain and the other six genes from A/PR/8/34.
7.13
Reverse genetics, as the term is used in molecular virology, describes the generation
of viruses possessing a genome derived from cloned DNA. Reverse genetics allows
for rapid and rational generation of influenza strains as each of the eight viral
segments can be derived from different influenza strains and recombined into a virus
with the desired reassortment of genes.
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Description of genetic modification
E. coli
7.14
The genetic modifications to non-pathogenic laboratory strains of E. coli (described in
Table 1) are not expected to increase the pathogenicity, virulence or infectivity of the
organisms to laboratory personnel, the community, or the environment. In addition,
the developments will not result in the organisms having a greater ability to escape
from containment than the unmodified organisms. No transcription or translation of
the cloned influenza cDNA will occur in the host E. coli strains and therefore no
infectious viral particles or viral proteins will be produced.
Mammalian cell lines
7.15
The cultured mammalian cell lines will be used as host cells to produce and replicate
live influenza viral particles as described in Table 1. The containment level required
for the genetic modification of the mammalian cell lines will depend on the risk
categorisation of the influenza virus being created. The genetically modified virus
strains produced by the mammalian cells and the Physical Containment (PC) level
required for containment of these activities are described below (sections 7.16-7.35).
Seasonal influenza
7.16
While influenza strains developed using reverse genetics have not been categorised in
the AS/NZS 2243.3:2002, the Committee considered that their Risk Group
classification will be dependent upon the progenitor influenza strains used, and the
combination of gene segments in the resultant virus.
7.17
The influenza strain A/PR/8/34 and other influenza stains commonly used in the
production of human influenza vaccines (vaccine strain) are Risk Group 2 viruses as
they can cause human or animal disease, but are unlikely to be a serious hazard to
laboratory workers, the community, livestock, or the environment. Reverse genetics
to develop a vaccine strain such as A/PR/8/34 would result in a virus with an identical
genome and characteristics to the progenitor strain and would also be a Risk Group 2
organism.
7.18
The seasonal influenza viruses are categorised as Risk Group 2 organisms as they can
cause human, or animal disease, but are unlikely to be a serious hazard to laboratory
workers, the community, livestock, or the environment. While laboratory exposures
may cause infection, effective treatment and preventive measures are available and
the risk of spread is limited. The strains developed by reverse genetics will contain
the HA and NA/NB gene sequences from the seasonal viruses and the remaining six
gene segments from an influenza vaccine stain. Site-directed mutagenesis of the HA
and/or NA gene sequences may occur for research purposes, to attenuate the virus, or
to create an apathogenic strain. For example, gene sequences that influence viral
growth may be altered to ensure that the virus can no longer survive in the lower
respiratory tract where it can cause serious illness (cold-adapted viruses). Therefore
these genetically modified organism could be classed as Risk Group 2 organisms.
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7.19
The reverse genetic development of the seasonal influenza strains involves the
introduction of nucleic acid that is characterized to the extent that its sequence is
known, its gene function is understood and its potential gene products are understood.
The modification is not expected to increase the pathogenicity, virulence or infectivity
of the organisms to laboratory personnel, the community, or the environment. In
addition, the developments will not result in the organisms having a greater ability to
escape from containment than the unmodified organisms.
Apathogenic strains of avian influenza
7.20
Pathogenic avian influenza strains are categorised as exotic animal viruses that must
be held under quarantine conditions at the CSIRO Australian Animal Health
Laboratory or other facilities approved by AQIS (Australia Quarantine and Inspection
Service) for these organisms (AS/NZS 2243.3:2002). These organisms may cause
serious disease in humans or animals and may present a serious hazard to laboratory
workers. However, only apathogenic strains of avian influenza will be derived by
reverse genetics. The viruses to be developed will be rendered apathogenic by sitedirected mutagenesis of pathogenicity determinants, for example the polybasic region
of the HA gene. The avian viruses may also be modified to facilitate growth or
attenuate the virus.
7.21
The applicant has noted that in an application for importation of genetically modified
apathogenic strains of influenza A virus that contained the H5 and N1 genes in a
A/PR/8/34 background genome (GMC06001), the head of the Australian/New
Zealand Standards Authority suggested that PC3 containment would be suitable for
the containment of apathogenic strains of avian influenza generated by reverse
genetics. For the current application, the Committee considered that these viruses
should also be held and manipulated at a minimum of PC3 containment.
7.22
The development of apathogenic avian influenza strains involves a microorganism of
Risk Group 3 or above and the development will result in genomes or fragments of
genomes capable of causing disease in humans, animals, plants, or fungi.
Containment regime
E. coli
7.23
The Committee considered that the experiments proposed in this application, to
develop genetically modified non-pathogenic laboratory strains of E. coli, should be
contained within a minimum of Physical Containment level 1 (PC1).
7.24
The facility to be used shall be approved and registered as a containment facility
under section 39 of the Biosecurity Act, in accordance with the MAF Biosecurity
New Zealand/ERMA New Zealand Standard Facilities for Microorganisms and cell
cultures 2007 (the Standard). This containment regime contains clear guidelines for
the safe handling and disposal of cultures.
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Influenza and mammalian cell lines
7.25
The appropriate containment regime for the influenza viruses developed by reverse
genetics, as well as for the mammalian cell lines that are used to create or maintain
these viruses, is dependent on the influenza strain involved.
Seasonal influenza
7.26
Seasonal influenza viruses are categorised as Risk Group 2. While, seasonal
influenza viruses developed using reverse genetics have no Risk Group designation
under AS/NZS 2243.4:2002, the Committee noted that the re-creation of seasonal
influenza strains using reverse genetics is not likely to increase the pathogenicity, host
range or transmission characteristics of the influenza viruses.
7.27
The applicant proposes to develop the seasonal strains of influenza in PC2
containment at the WHO National Centre for Influenza at ESR. The applicant notes
that the standard operating procedures in place include procedures for handling of all
infectious material in closed, sealed containers or within class II biological safety
cabinets, decontamination protocols and training requirements. The applicant notes
that these standard operating procedures will be followed for the development of
seasonal strains of influenza by reverse genetics.
7.28
The Committee noted that PC2 containment is used for seasonal influenza
surveillance and for reverse genetic manipulation of seasonal influenza viruses in
other laboratories around the world. The Committee also noted that the WHO
National Centre for Influenza, handles nearly 800 seasonal influenza virus isolates
every year during routine surveillance and there have been no laboratory acquired
infections to date. This surveillance work is carried out in PC2 containment, within a
facility registered to the Standard. ESR has procedures in place to minimise the risk
of escape from containment and infection of laboratory personnel.
7.29
The Committee considered that the genetically modified seasonal strains of influenza
should be developed in a containment facility registered under the Biosecurity Act
1993 in accordance with the Standard and operated at a minimum of PC2 (control 1.2
in Appendix 1). This containment regime contains clear guidelines for the safe
handling and disposal of cultures.
Apathogenic strains of avian influenza
7.30
The applicant states that the reverse genetic development of apathogenic strains of
avian influenza will be undertaken solely at the PC3 laboratory situated at the
Wallaceville Research Centre. This facility is shared between MAF Investigation and
Diagnostic Centre (IDC) and ESR, as part of the National Centre for Biosecurity and
Infectious Disease (NCBID).
7.31
The Committee considered that the genetically modified apathogenic strains of avian
influenza should be developed in a containment facility registered under the
Biosecurity Act 1993 in accordance with the Standard and operated at PC3 (control
1.2 in Appendix 1). The minimum requirements for PC3 containment are specified in
the AS/NZS 2243.3:2002.
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7.32
The Committee noted that ESR has signed a lease with the MAF IDC to use the PC3
facility at Wallaceville, Upper Hutt. This lease specifies that ESR will follow all
health and safety procedures and relevant standard operating procedures in place at
this facility.
7.33
The Committee noted that this PC3 facility is located in a building that is designed for
maximum earthquake resistance, a category 1 structure, the highest category in the
structure design code. All windows to the facility are sealed and made from doubleglazed laminated safety glass that is strengthened to reduce the chance of a
containment breach through accidental breakage.
7.34
The Committee noted that IDC’s PC3 facility has been operating well since it was
established in 2000. This facility has good standard operating procedures in place to
satisfy the requirements in relation to PC3 operations.
7.35
In summary, the Committee considered that the containment regime described above
will be effective in containing the genetically modified organisms. The Committee
considered that a laboratory registered to the Standard and operated at a minimum of
PC1 for developments involving of non-pathogenic strains of E. coli, PC2 for the
developments involving seasonal influenza and PC3 for developments involving
apathogenic avian influenza is an appropriate containment regime (control 1.2). The
Committee noted that the Standard requires facilities to be constructed and operated in
a manner to ensure organisms are securely contained and held only within the facility.
The provisions in the Standard that ensure that containment is maintained cover
access to the facility, staff training, contingency plans, waste disposal, record keeping
and packaging for organisms in transit.
Potential pathways for escape of organisms from the containment facility
7.36
The Committee considered the following potential pathways of escape of the
genetically modified organisms:
i)
Escape during transport.
ii)
Escape from containment facilities by intentional removal by staff or
unauthorized persons.
iii)
Escape from containment facilities by accidental or unintentional removal by
staff or unauthorised persons, for example:
iv)

Via infection of personnel working with the microorganisms;

Through passive vectoring (eg on clothing or skin of personnel);

Contaminated liquid or solid waste leaving the containment
facility;

Through air discharged from the containment facility.
Escape from containment following natural disaster (such as flood and
earthquakes) or fire.
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7.37
The Committee concluded that escape of the organisms via pathways i - iv is highly
improbable for all organisms as described in Table 1.
7.38
This conclusion was formed on the basis of the provisions of the Standard and the
AS/NZS 2243.3:2002 imposed by control 1.2 that relate to packaging of the
organisms for transport, waste disposal procedures, the management of the facility
(including access to the facility and staff training) and the requirement for
contingency plans. To enhance staff training, the Committee has imposed control 1.3
requiring the person responsible for the operation or particular research area of the
facility to ensure that all staff are aware of the containment controls on this approval.
This conclusion was also formed based on the strict laboratory operating procedures
and the controls imposed in Appendix 1.
7.39
The Committee noted that all organisms must be transported to and within New
Zealand in packages complying with the packaging (Packing Instruction No 602) and
labelling requirements of the most recent version of the IATA Dangerous Goods
Regulations and the packaging requirements of the Standard and AS/NZS
2243.3:2002.
7.40
In the case of the development of apathogenic strains of avian influenza, the
ventilation system and the laboratory procedures in a PC3 facility ensure that all air,
infectious waste, equipment and clothing is decontaminated in accordance with the
requirements of the AS/NZS 2243.3:2002, before leaving the containment facility.
7.41
The Standard requires contingency plans to be in place for use in the event of
accidental release of new organisms outside the facility and for fire and other
emergencies. Section 8.9 Contingency Plans, states “…In the event of any spillage or
breach of containment of an organism, the contingency plan must be implemented
immediately”. The Committee has imposed controls to enhance this measure
requiring the contingency plan to be implemented immediately following any breach
of containment (control 5.3) and notification to the MAF Inspector of that facility
following such an occurrence (control 5.2).
Conclusion on adequacy of the containment regime
7.42
The Committee considered the biological characteristics of the genetically modified
strains of E. coli, mammalian cell lines, seasonal influenza and apathogenic avian
influenza, the containment conditions, and the potential pathways of escape. Taking
all of these into consideration the Committee conclude that it is highly improbable
that the genetically modified strains of E. coli, mammalian cell lines, seasonal
influenza and apathogenic strains of avian influenza would be able to escape from
containment.
8
Ability of the organisms to establish a self-sustaining
population and ease of eradication
8.1
In accordance with sections 44 and 37 and clause 10(e), the Committee considered the
ability of genetically modified strains of E. coli, mammalian cell lines, seasonal
influenza and apathogenic avian influenza strains developed by reverse genetics to
form a self-sustaining population should they escape from containment and the ease
of eradication of such populations.
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8.2
Based on the biological characteristics of the genetically modified strains of nonpathogenic E. coli, in particular the requirement for growth factors, such as amino
acid supplementation, it is highly improbable that the non-pathogenic laboratory E.
coli strains could survive outside of containment and form a self-sustaining
population.
8.3
The mammalian cells to be used in these experiments have specific growth
requirements that would limit their ability to survive outside of containment. These
cells require a controlled atmosphere that is enriched for CO2 and immersion in cell
culture media for survival. Mammalian cell lines are easily exposed to, and destroyed
by opportunistic microbial agents, or can easily be destroyed by deprivation of growth
media. It is highly improbable that the mammalian cell lines could survive outside
of containment and form a self-sustaining population.
8.4
Based on the biological characteristics of the influenza viruses to be developed, in
particular their limited ability to survive outside of host cells for long periods, The
Committee considered it highly improbable that a self-sustaining population would
form. The development of a self-sustaining population would depend on an escape
from containment occurring (assessed as highly improbably in section 7.36), a
susceptible host being in the vicinity of the containment facility, and that susceptible
host being exposed to a dose of viable pathogen sufficient to lead to an infection. In
order to establish a self-sustaining population, an infected host would then need to
come into contact with, and transmit virus to, other susceptible hosts. There is some
uncertainty as to the range of susceptible hosts, the seasonal influenza viruses
developed are likely to be able to infect humans, whereas the avian derived influenza
viruses developed are likely to preferentially infect avian species and only infect
humans exposed to extremely large viral doses. There is further uncertainty around
how infectious the apathogenic avian influenza viruses would be. However, as the
apathogenic avian influenza viruses developed will contain avian HA and NA gene
segments, it is considered likely that transmission may occur between avian species.
9
Identification and assessment of potentially significant
adverse and beneficial effects (risks, costs and benefits) –
9.1
The potential risks, costs and benefits assessed here are those identified as significant,
having regard for those matters set out in clauses 9 and 10 of the Methodology, which
reflect sections 5, 6, 8 and 44 of the Act. Risks were considered in terms of the
requirements of section 45(4) of the Act and clause 12 of the Methodology, including
the assessment of consequences and probabilities, the impact of uncertainty and the
impact of risk management. The potentially significant adverse effects were
categorised and considered in terms of their area of impact on the environment, on
human health and safety, Māori and their culture and traditions, the market economy
and society and the community. Costs and benefits were considered in terms of
clause 13 of the Methodology. A “cost” is defined in clause 2 as “the value of a
particular adverse effect expressed in monetary or non-monetary terms”. Therefore,
these have been assessed in an integrated fashion together with the risks of those
adverse effects in the following assessment.
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9.2
The Committee considered that, given the biological characteristics of the nonpathogenic laboratory strains of E.coli, the seasonal strains of influenza generated by
reverse genetics and the mammalian cells used to generate the seasonal strains of
influenza, the proposed containment system and controls (see Appendix 1), there is no
evidence for, nor any reason to expect, any non-negligible adverse effects on humans,
animals, the environment, the society and community or the market economy.
9.3
The Committee considered that there are no adverse effects to the market economy or
the society and community of approving the development in containment of
genetically modified apathogenic strains of avian influenza
9.4
The Committee have assessed the potential risks, costs and benefits of the
development of apathogenic strains of avian influenza to the environment, human
health and safety and Māori culture and traditions below.
The environment
Potential for the strains of avian influenza developed by reverse genetics to be
pathogenic to native and valued animals in New Zealand
9.5
The Committee considered the potential for the genetically modified strains of avian
influenza to be pathogenic to native and valued animals in New Zealand. The
Committee noted that the avian influenza viruses developed will apathogenic.
Therefore the risk to the environment would depend on the ability of the virus to
revert to a pathogenic form.
9.6
The Committee noted that it is difficult to determine the magnitude of effect of the
viruses reverting to a pathogenic form through antigenic drift or antigenic shift.
However, the strains of avian influenza to be developed will be made apathogenic
through removal or alteration of pathogenic sequences in the HA gene. One
modification which is commonly performed to render avian influenza apathogenic is
the deletion of the polybasic region of the HA, such deletions have been demonstrated
to be stable over serial passaging (GMC06001). However, the Committee noted that
for this risk to eventuate the virus must first escape and form a self-sustaining
population, of which the likelihood of which has been previously assessed as highly
improbable (section 7.40). Therefore, this adverse effect was assessed as negligible
given the proposed containment regime and controls.
Human health and safety
Potential for strains of avian influenza generated by reverse genetics to be
pathogenic to humans
9.7
The Committee considered the potential for the genetically modified apathogenic
strains of avian influenza viruses generated by reverse genetics to be pathogenic to
humans.
9.8
The Committee noted that should an infection occur, the effects would be minimal as
the virus is apathogenic. The Committee also noted that, if the virus remains in
containment, the primary potential for an infection to occur would be through
occupational exposure of laboratory workers. However, human infection is
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highly improbable due to the biological characteristics of the virus, including the
host cellular receptor specificity of avian influenza (sections 7.9-7.10), the use of
safety equipment such as biological safety cabinets, the use of protective clothing
such as gowns, face masks and gloves, and extensive training and compliance with
safety procedures. Based on the magnitude and likelihood of the effect the level of
risk is A. The Committee concluded that the adverse effect is negligible.
Māori culture and traditions
9.9
The Committee considered the potential Māori cultural effects of these applications in
accordance with clauses 9(b)(i) and 9(c)(iv) of the Methodology and sections 6(d) and
8 of the Act. In addition, the Committee used the assessment framework contained in
the ERMA New Zealand User Guide “Working with Māori under the HSNO Act
1996”, and the ERMA New Zealand revised protocol “Incorporating Māori
perspectives in Part V Decision-making”, as guides in assessing the information
contained in these applications.
9.10
The Committee noted no native DNA samples from taonga species are used in this
application and no use of human DNA derived from Māori also.
9.11
The Committee noted also that no significant issues were identified between the
applicant and local iwi/Māori (Tenths Trust) during informal communication
regarding this application.
9.12
Taking into account the assessment of the potential adverse environmental effects
associated with this application, the Committee considers that this application
presents negligible risk to Māori culture or traditional relationships with ancestral
lands, water, sites, wāhi tapu, valued flora and fauna or other taonga.
9.13
This assessment is made on the condition that the specimens are, transported, handled,
stored, and used in accordance with all proposed controls, conditions and relevant
regulations.
Beneficial effects
9.14
The Committee considered the potential beneficial effects associated with this
application, in accordance with sections 5 and 6(e) of the Act and clauses 9, 10, 13,
and 14 of the Methodology.
9.15
The Committee considered that there are no beneficial effects to the environment, the
market economy or human health and safety of approving the development in
containment of genetically modified apathogenic strains of avian influenza.
9.16
The Committee noted that the purpose of this research is to create influenza viruses by
reverse genetics in laboratory containment, for future application in diagnostics,
research and vaccine manufacture. The Committee noted that for future application
of these viruses, for example in vaccine manufacture, an additional HSNO Act
approval would be required. Potential benefits arising from the future application of
the viruses developed by reverse genetics have not been considered as they fall
outside the scope of this approval.
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Society and community
Upskilling of the workforce and enhancement of scientific knowledge
9.17
The Committee noted that the development of apathogenic strains of avian influenza
would lead to upskilling of the workforce, would increase scientific knowledge in
regards to influenza and other viruses, and would enhance the ability of New Zealand
to respond to an influenza epidemic. The magnitude of this effect is considered by the
Committee to be minor and it is very likely to eventuate. Thus the level of beneficial
effect is E. The Committee concluded that this benefit is non-negligible.
10
Overall evaluation of risk, costs and benefits
Precautionary approach
10.1
Section 7 of the Act requires the Committee to take into account the need for caution
in managing adverse effects where there is scientific and technical uncertainty about
those effects. The Committee used scenarios to set upper and lower bounds on the
assessment of risks and the evaluation was based on the higher value of the risk.
Clause 29 of the Methodology notes that where there is scientific and technical
uncertainty the Authority must considered the materiality of the uncertainty to the
decision. Since none of the risks was assessed as being non-negligible, the
Committee concluded that this uncertainty was not material to the decision.
Approach to risk
10.2
Clause 33 of the Methodology requires the Authority to have regard for the extent to
which a specified set of risk characteristics exist when considering applications. This
provision provides a route for determining how cautious or risk averse the Authority
should be in weighing up risks and costs against benefits. In the present applications
clause 33 is influenced by the applications being “in containment” and the conclusion
that the containment provisions and controls will reduce most biological and physical
risks to a low level.
10.3
The Committee considered that as the identified biological, physical, or human health
risks were assessed as being negligible, caution in addition to that required by section
7 of the Act is not warranted.
Aggregation and comparison of risks, costs and benefits
10.4
The overall evaluation of risks, costs and benefits was carried out in accordance with
section 45 of the Act and clause 26 of the Methodology, having regard to clauses 22
and 34 of the Methodology.
10.5
The adverse effects identified by the Committee were the potential of the apathogenic
strains of avian influenza developed using reverse genetics to be pathogenic to native
and valued animals in New Zealand, or be pathogenic to humans. The Committee
assessed these adverse effects and concluded the risks to be negligible if the proposed
containment regime and the controls in Appendix 1 are adhered to.
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10.6
The significant beneficial effects identified by the Committee were the upskilling of
the workforce and enhancement of scientific knowledge. The Committee assessed
these benefits and concluded that the benefits are non-negligible.
10.7
The Committee was unable to find common units of measurement with which to
combine risks, costs, and benefits in accordance with clause 34(a) and there were no
dominant sources of risk (clause 34(b)). Because the risks individually and as a
whole are negligible, the decision is made in accordance with clause 26 (not clause
27) of the Methodology.
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11
Decision
11.1
Pursuant to section 45(1)(a)(i) of the Act, the Committee is satisfied that this
application is for a purpose specified in section 39(1) of the Act, namely section
39(1)(a) of the Act: “the development of any new organism.”
11.2
The Committee is satisfied that the containment regime, as set out in Appendix 1 of
this decision, will adequately contain the organisms as required by section
45(1)(a)(iii) of the Act.
11.3
The Committee evaluated the potential of genetically modified organisms (as
described in Table 1) to escape from containment in accordance with section 44(b) of
the Act. Having considered the proposed containment regime, the biological
characteristics of the organisms and the potential pathways for escape from
containment, the Committee concluded that it is highly improbable that the
organisms would escape from containment.
11.4
In accordance with section 37 of the Act, the Committee evaluated the potential of the
genetically modified organisms to establish an undesirable self-sustaining population
should they escape containment. The Committee considered that it is highly
improbable that an undesirable self-sustaining population could establish. In the
event that a population did establish, they would be difficult to detect and almost
impossible to eradicate.
11.5
Having considered all the possible effects in accordance with sections 45(1)(a)(ii),
45(4) and 44 and pursuant to clause 26 of the Methodology, and based on
consideration and analysis of the information provided and taking into account the
application of risk management controls specified in Appendix 1 of this decision, the
view of the Committee is that the risks (or costs) of adverse effects associated with
the development of influenza viruses by reverse genetics are outweighed by the
benefits.
11.6
In accordance with clause 36(2)(b) of the Methodology the Committee records that, in
reaching this conclusion, it has applied the balancing tests in section 45 of the Act and
clause 26 of the Methodology and has relied in particular on the criteria set out in the
following sections of the Act:

section 44 additional matters to be considered;

section 45 determination of application;

section 37 additional matters to be considered; and

the Third Schedule (Part I) matters to be addressed by containment controls for
importing, developing or field testing of genetically modified organisms.
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11.7

The Committee has also applied the following criteria in the Methodology:

clause 9 – equivalent of sections 5, 6 and 8;

clause 10 – equivalent of sections 36 and 37;

clause 12 – evaluation of assessment of risks;

clause 13 – evaluation of assessment of costs and benefits;

clause 20 – information produced from other bodies;

clause 21 – the decision accords with the requirements of the Act and
regulations;

clause 22 – the evaluation of risks, costs and benefits – relevant considerations;

clause 24 – the use of recognized risk identification, assessment, evaluation and
management techniques;

clause 25 – the evaluation of risks;

clause 26 – the risks are negligible and it is evident benefits outweigh costs;

clause 29 and 32 – considering uncertainty;

clause 33 – the risk characteristics; and

clause 34 – the aggregation and comparison of risks, costs and benefits.
Application GMD06058 to develop in containment E. coli and mammalian cells lines,
seasonal influenza viruses and apathogenic avian influenza viruses by reverse
genetics, is thus approved, with controls, in accordance with section 45(1)(a) of the
Act. As required under section 45(2) the approval is subject to the controls listed in
Appendix 1 of this decision.
_____________________
2 October 2007
Kieran Elborough
Date
Chair New Organisms (GMO) Committee
Approval code (BCH code): GMD004745 – 49 (41040 – 41044)
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Approval numbers and BCH numbers for Organisms in Application GMD06058
Approval code
GMD004747
GMD004748
GMD004745
GMD004746
GMD004749
Organism
Escherichia coli (Migula 1895) Castellani &
Chalmers 1919 (GMD06058)
Homo sapiens (Linnaeus, 1758) (GMD06058)
Canis familiaris (Linnaeus, 1758) (GMD06058)
Chlorocebus aethiops (Linnaeus,1758)
(GMD06058)
Influenza A (Lamb & Krug, 2001) (GMD06058)
Environmental Risk Management Authority Decision: GMD06058
BCH number
41040
41041
41042
41043
41044
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Appendix 1: Controls Required by this Approval
In order to satisfactorily address the matters detailed in the Third Schedule Part I:
Matters to be Addressed by Containment Controls for Importing, Developing and
Field Testing of Genetically Modified Organisms4, of the Act, and other matters in
order to give effect to the purpose of the Act, the approved organisms are subject to
the following controls:
1
To limit the likelihood of any accidental release of any organism or any viable
genetic material5.
1.1 The approved organism shall be developed and maintained within a containment facility
which complies with these controls.
1.2 The construction, operation, and management of the microorganism containment facility
shall be in accordance with the:
1.2.1 MAF Biosecurity New Zealand and ERMA New Zealand Standard: Facilities
for Microorganisms and cell culture6.
1.2.2 Australian New Zealand Standard AS/NZS 2243.3: 20026 Safety in
Laboratories: Part 3: (Microbiological aspects and containment facilities),
excluding those deviations specified in the Standard listed in 1.2.1.
1.2.3 Physical Containment Level 1 (PC1) requirements of the above standards for
non-pathogenic strains of E. coli.
1.2.4 Physical Containment Level 2 (PC2) requirements of the above standards for
strains of seasonal influenza and the mammalian cell lines used to develop
seasonal influenza strains.
1.2.5 Physical Containment Level 3 (PC3) requirements of the above standards for
apathogenic strains of avian influenza and the mammalian cell lines used to
develop apathogenic strains of avian influenza strains.
1.3 The person responsible for the operation of the containment facility shall inform all
personnel involved in the handling of the organisms of the Authority’s controls.
1.4 The containment facility shall be approved by MAF Biosecurity New Zealand, in
accordance with section 39 of the Biosecurity Act and the standards listed in control 1.2.
Bold headings refer to matters to be addressed by containment controls for [importing, developing or
field testing] of genetically modified new organisms, specified in the Third Schedule (Part I) of the HSNO
Act 1996.
5 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, e.g. when organisms or parts thereof are sub-lethally damaged by being frozen, dried, heated, or
affected by chemical.
6 Any reference to this standard in these controls refers to any subsequent version approved or endorsed
by ERMA New Zealand.
4
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2
To exclude unauthorised people from the facility:
2.1 The identification of entrances, numbers of and access to entrances, and the security
requirements for the entrances and the facility shall be in compliance with the standards
listed in control 1.2.
3
To exclude other organisms from the facility and to control undesirable and
unwanted organisms within the facility:
3.1 The exclusion of other organisms from the facility and the control of undesirable and
unwanted organisms within the facility shall be in compliance with the standards listed in
control 1.2.
4
To prevent unintended release of the organism by experimenters working with
the organism:
4.1 The prevention of unintended release of the organisms by experimenters working with
the organisms shall be in compliance with the standards listed in control 1.2.
5
To control the effects of any accidental release or escape of an organism:
5.1 Control of the effect of any accidental release or escape of the organism shall be in
compliance with the standards listed in control 1.2.
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.
5.3 In the event of any breach of containment of the organism, the contingency plan for the
attempted retrieval or destruction of the organism that has escaped shall be implemented
immediately. The contingency plan shall be included in the containment manual in
accordance with the requirements of the standards listed in control 1.2.
6
Inspection and monitoring requirements for containment facilities:
6.1 The operation of the containment facilities shall comply with the requirements contained
in the standards listed in control 1.2 relating to the inspection and monitoring
requirements for containment facilities.
6.2 The containment manual shall be updated, as necessary, to address the implementation of
the controls imposed by this approval, in accordance with the standards listed in control
1.2.
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7
Qualifications required of the persons responsible for implementing these
controls:
7.1 The training of personnel working in the facility shall be in compliance with the
Standards listed in control 1.2.
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