ENVIRONMENTAL RISK MANAGEMENT
AUTHORITY DECISION
Amended under s67A on 16 August 2007
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27 February 2006
GMD06004
 To develop in containment genetically modified organisms
under sections 40(1)(b) and 42A of the Hazardous Substances
and New Organisms (HSNO) Act 1996.
Victoria University of Wellington
Applicant
To develop in containment genetically modified bacteria to
Purpose
study primary and secondary metabolism and virulence
mechanisms in fluorescent Pseudomonas species.
20 17 February 2006
Date received
Consideration date 22 27 February 2006
Chief Executive, ERMA New Zealand
Considered by
Application code
Application type
1 Summary of decision
1.1 The application to develop, as a project, genetically modified organisms in
containment is approved, with controls, having been considered in
accordance with the relevant provisions of the Hazardous Substances and New
Organisms (HSNO) Act 1996 (the Act), the HSNO (Low-Risk Genetic
Modification) Regulations 2003 (the Low-Risk Regulations), and the HSNO
(Methodology) Order 1998 (the Methodology).
The organisms approved are:
1.2 The organisms approved for development are described in Table 1:
Table 1: Organisms as recorded on ERMA New Zealand Register
Host organism
Category
Modified by:
of host
organism
Escherichia coli (Migula
1895) Castellani &
Chalmers 1919
non-pathogenic laboratory
strains
1
Pseudomonas aeruginosa
(Schroeter 1872) Migula
1900. Strain PAO1
(sequenced; Stover et al.,
2000)
2
Pseudomonas fluorescens
Migula 1895
1
Pseudomonas putida
(Trevisan 1889) Migula
1895
1
Pseudomonas syringae van
Hall 1902.
Pseudomonas syringae pv.
phaseolicola (Joardar et
al., 2005)
2
Site directed mutagenesis of genes encoding the
following types of proteins:
 Quinone oxidoreductase enzymes
 Non-ribosomal peptide synthetase and
polyketide synthetase enzymes
 Stress-sensing and regulatory proteins
 DNA replication and modification
proteins
 Protein and peptide secretory proteins
 Surface proteins, adhesins and antigens
 Small molecule and macromolecule
metabolism proteins.
Standard non-conjugative Escherichia coli and
Pseudomonas cloning and expression plasmid
vectors containing genomic DNA and PCR
products from, E. coli, P. aeruginosa, P. putida,
P. syringae, P. fluorescens, or P. stutzeri.
Donor DNA derived from the above
microorganisms will contain regulatory
elements, non-coding regions and genes
encoding proteins belonging to the following
functional families of proteins:
 Quinone oxidoreductase enzymes
 Non-ribosomal peptide synthetase and
polyketide synthetase enzymes
 Stress-sensing and regulatory proteins
 DNA replication and modification
proteins
 Protein and peptide secretory proteins
 Surface proteins, adhesins and antigens
 Small molecule and macromolecule
metabolism proteins
Additional donor DNA will include nonribosomal peptide synthetase genes derived
from non-pathogenic bacterial hosts.
Vector DNA will also include standard
inducible promoters, selectable markers, protein
purification tags, E.coli and Pseudomonas
origins of replications.
Genetic material shall not include genes
encoding known bacterial toxins.
Environmental Risk Management Authority Decision: GMD06004
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Category of
modification/
containment
level
A/PC1
B/PC2
A/PC1
A/PC1
B/PC2
2 Legislative Criteria for Application
2.1
The application was lodged pursuant to section 40(1)(b) of the Act and
determined according to the rapid assessment provisions of section 42A of
the Act.
2.2
The application has been approved with controls by Mr Rob Forlong, Chief
Executive of ERMA New Zealand, under delegation from the Authority as
provided for in section 19 of the Act.
3 Consideration
Sequence of the consideration
3.1
The application was formally received and verified as containing sufficient
information on 17 February 2006.
3.2
The decision was based on the information supplied by the applicant in their
application form: Develop in containment a project of low risk genetically
modified organisms by rapid assessment (ER-AF-NO3P-1).
3.3
The application was considered by the Chief Executive of ERMA New
Zealand. Relevant staff within ERMA New Zealand, including the Acting
Manager Māori, were involved in providing advice on the consideration of
the application.
3.4
The development of the genetically modified organism described above
(Table 1) meets the criteria of a low-risk genetic modification specified in
the Regulations made under section 41 of the Act, being the HSNO (LowRisk Genetic Modification) Regulations 2003.
3.5
In reaching my decision I have used information that is relevant and
appropriate to the scale and significance of the risks, costs, and benefits
associated with the genetic modifications and matters relevant to the purpose
of the Act, as specified in Part II, and followed the relevant provisions of the
Methodology.
3.6
In accordance with section 42A of the Act for rapid assessment, the
approach adopted was to identify the circumstances of the genetic
modification, to evaluate these against the criteria specified in section 41 of
the Act, and to consider whether there are any residual risks that require
further consideration. This approach covered the following issues:
 Purpose of the application (section 39 of the Act)
 Assessment against the criteria for low-risk genetic modification
(section 42A of the Act)
 Identification and assessment of the risks and other impacts of the
organism
 Precedents
 Proposed controls
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3.7
The Department of Conservation (DoC) was notified upon receipt of this
application.
3.8
DoC responded with comments on the application by email on 24 February
2006 , and concluded with the following statement:
“The Department considers that it is extremely unlike that the proposed
modification will increase the ability of the host organisms to escape
containment. Therefore, it considers that if the above containment conditions
are properly implemented the likelihood of escape of a developed organism
to be very low.
Based on that likelihood, the Department considers that the overall risk the
developed organisms poses to its mission to be negligible.”
Purpose of the application
3.9
Pseudomonas aeruginosa is an important opportunistic pathogen of humans,
and is inherently resistant to antibiotics. Consequently, there is much
interest in identifying “virulence factors”, enzymes and other
macromolecules which assist this organism in causing disease, and which
are potential targets for novel therapeutic agents.
3.10 The primary aim of this research is to study genes and enzymes involved in
virulence of P. aeruginosa and to compare these to similar genes and
enzymes in the related fluorescent bacteria Pseudomonas putida,
Pseudomonas fluorescens, and Pseudomonas syringae. These genes encode
the following types of proteins:
 Quinone oxidoreductase enzymes
 Non-ribosomal peptide synthetase and polyketide synthetase enzymes
 Stress-sensing and regulatory proteins
 DNA replication and modification proteins
 Protein and peptide secretory proteins
 Surface proteins, adhesins and antigens
 Small molecule and macromolecule metabolism proteins.
3.11 The function of these genes and enzymes will be determined on the basis of
their expression and biochemical properties and also by creating targeted
gene knockouts, and over-expressing specific genes in the four
Pseudomonas species, and E. coli. The applicant states that understanding of
the mechanisms by which these enzymes promote virulence will open the
way for design of novel therapeutic agents. A further beneficial outcome is
that some of these enzymes may also have applications in anti-cancer gene
therapy.
3.12 I have determined that this application is for a valid purpose being the
development of any [new] organism as provided for in section 39(1)(a) of
the Act.
Environmental Risk Management Authority Decision: GMD06004
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Assessment against the criteria for low-risk genetic
modification
3.13 Category of host organisms:
The established laboratory strains of Pseudomonas fluorescens,
Pseudomonas putida and non-pathogenic laboratory strains of Escherichia
coli to be used by the applicant are not capable of causing disease in
humans, animals, plants or fungi nor do they produce desiccation-resistant
structures, such as spores or cysts. As such, these bacterial strains are
considered Category 1 host organisms as defined in clause 7(1) of the HSNO
(Low-Risk Genetic Modification) Regulations 2003.
Pseudomonas aeruginosa and Pseudomonas syringae pv phaseolicola are
capable of causing disease in humans, animals, plants or fungi and as such,
are considered Category 2 host organisms as defined in clause 7(2) of the
HSNO (Low-Risk Genetic Modification) Regulations 2003. P. aeruginosa is
an opportunistic pathogen and the applicant proposes to use a genomesequenced laboratory strain, P. aeruginosa PAO1, which is much less
virulent than other clinical isolates. P. syringae pv phaseolicola is generally
non-pathogenic except to a variety of beans, on which it can cause halo
blight. The genome of this organism has also been sequenced.
3.14 Category of genetic modification:
The proposed genetic modifications to established laboratory strains of P.
fluorescens, P. putida and non-pathogenic laboratory strains of E. coli 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.
Therefore, the genetic modifications as described in Table 1 of this decision
are Category A genetic modifications as defined in clause 5(1) of the HSNO
(Low-Risk Genetic Modification) Regulations 2003 and require a minimum
of Physical Containment Level 1 (PC1).
The proposed genetic modification on Category 2 hosts, P. aeruginosa
PAO1 and P. syringae pv phaseolicola involve characterised nucleic acids
in which the sequence is known, gene function and potential gene products
understood. These modifications are not expected to increase the
pathogenicity, virulence or infectivity of the organisms to laboratory
personnel, the community, or the environment. In addition, these
developments will not result in the organisms having a greater ability to
escape from containment than the unmodified organisms. Therefore, the
genetic modifications as described in Table 1 of this decision are Category B
genetic modifications as defined in clause 5(2) of the HSNO (Low-Risk
Genetic Modification) Regulations 2003 and require a minimum of Physical
Containment Level 2 (PC2).
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3.15 I am satisfied that the development meets the criteria for low-risk genetic
modification specified in the Regulations, made under section 41 of the Act.
The experiments on P. fluorescens, P. putida and E. coli meet the
requirements of Category A modifications as defined in clause 5 of the
Regulations in that the modification involves a category 1 host organism and
is to be carried out under a minimum of PC1 containment. Experiments on
P. aeruginosa PAO1 and P. syringae pv phaseolicola meet the requirements
of Category B modifications and is to be carried out under a minimum of
PC2 containment.
Identification and assessment of the risks, costs and other
impacts of the organism
3.16 I consider that the information provided by the applicant is relevant and
appropriate to the scale and significance of the risks, costs, and benefits
associated with the application (as required by clause 8 of the
Methodology). In accordance with clauses 9 and 10 of the Methodology the
information supplied by the applicant has been evaluated as follows:
3.17 I consider that, given the controls attached to this approval, there is no
evidence for, nor any reason to expect, any non-negligible adverse effects of
the proposed genetically modified organism on humans, animals, plants,
other organisms or the environment.
3.18 I have considered the potential Māori cultural effects in accordance with
sections 6(d) and 8 of the Act and clauses 9(b)(i), 9(c)(iv) of the
Methodology, in consultation with the Acting Manager, Māori.
3.19 As this application does not involve the use of genetic material from native
or valued flora and fauna from New Zealand or DNA sourced from Māori,
and as this application is for a development in containment, there is no
requirement for the applicant to consult with Māori.
3.20 Although recognising that iwi/Māori maintain an ongoing interest and
concern in the potential long term cultural implications of genetic
modification generally, I consider that this application poses negligible risk
of adverse effects to the relationship of Māori culture and traditions with
their ancestral lands, water, sites, waahi tapu, valued flora and fauna, and
other taonga.
Precedents
3.21 I must consider each application on its merits, and am therefore not bound
by the stance taken in previous decisions. However, in reflecting on
previous decisions that involved similar genetic modifications to those
proposed by this application, I note that low-risk genetic developments of
non-pathogenic E. coli, and P. aeruginosa, P. fluorescens, P. putida and P.
syringae have been considered and approved on many occasions by
Institutional Biological Safety Committees (IBSCs) under delegated
authority and by the Chief Executive of ERMA New Zealand.
Environmental Risk Management Authority Decision: GMD06004
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3.22 I consider that this current application does not raise any novel issues that
would warrant it not to be considered via section 42A of the Act.
Containment
3.23 The proposed genetic modifications on category 1 hosts, E. coli, P.
fluorescens and P. putida, meet the requirements of Category A genetic
modifications as defined in clause 5 of the HSNO (Low-Risk Genetic
Modification) Regulations 2003. Category A experiments are required to be
contained within a Physical Containment level 1 facility (PC1) registered
under MAF/ERMA New Zealand Standard 154.03.02 ‘Containment
Facilities for Microorganisms’. The proposed genetic modifications on
Category 2 hosts, P. aeruginosa PAO1 and P. syringae pv phaseolicola
meet the requirements of Category B modifications and is to be contained
within Physical Containment level 2 (PC2) of the above standard. The
containment regime contains clear guidelines for the safe handling and
disposal of bacterial cultures.
3.24 The facility in which the organisms will be maintained shall comply with
the requirements of the Australian New Zealand Standard AS/NZS
2243.3:2002 Safety in Laboratories: Part 3: Microbiological aspects of
containment and facilities, except for the deviations specified in the
MAF/ERMA New Zealand Standard 154.03.02. The laboratory proposed to
be used by the applicant is currently approved and registered as a
containment facility under section 39 of the Biosecurity Act, in accordance
with the MAF/ERMA New Zealand Standard 154.03.02.
4 Decision
4.1
I am satisfied that this application is for one of the purposes specified in
section 39(1) of the Hazardous Substances and New Organisms Act 1996,
being section 39(1)(a): the development of any [new] organism.
4.2
Based on consideration and analysis of the information provided, and having
considered the characteristics of the organism that is the subject of this
approval, the modification and the criteria for low-risk genetic modification
detailed in the HSNO (Low-Risk Genetic Modification) Regulations 2003, I
am of the view that the organism meets the criteria for rapid assessment
under section 42A of the Hazardous Substances and New Organisms Act
1996.
4.3
I am satisfied that the proposed containment regime and the controls
imposed in accordance with section 42A(3)(b) of the Hazardous Substances
and New Organisms Act 1996, as set out below, will adequately contain the
organism.
4.4
Pursuant to section 42A(3)(a) of the Hazardous Substances and New
Organisms Act 1996, and acting under delegation from the Authority
provided for in section 19 of the Act, I have approved this project
application for genetically modified non-pathogenic laboratory strains
Escherichia coli as described in Table 1 of this decision, subject to the
controls specified herein.
Environmental Risk Management Authority Decision: GMD06004
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4.5
In reaching this decision I have relied upon the following criteria in the Act
and the Methodology:
 Criteria for assessing the purpose of the application (section 39 of the
Act)
 Criteria for rapid assessment of adverse effects for the development of a
genetically modified organism in containment (section 42A of the Act).
 Criteria for a low-risk genetic modification specified in the HSNO
(Low-Risk Genetic Modification) Regulations 2003, made under section
41 of the Act.
 The information provided by the applicant was assessed against the
criteria in clauses 9, 10 and 12 of the HSNO (Methodology) Order 1998.
 Matters to be addressed by containment controls for developing
genetically modified organisms specified in Part 1 of the Third Schedule
to the Act.
5 Controls
In order to provide for the matters detailed in Part 1 of the Third Schedule of the Act1,
Containment Controls for Importation, Development and Field Testing of Genetically
Modified Organisms, and other matters in order to give effect to the purpose of the
Act, the approved organism is subject to the following controls:
1 To limit the likelihood of any accidental release of any organism or
any viable genetic material2.
1.1
The approved organism shall be developed and maintained within a containment
facility which complies with these controls.
1.2
The person responsible for a particular research area and/or the person
responsible for the operation of the containment facility shall inform all
personnel involved in the handling of the organism of the Authority’s controls.
1.3
The facility shall be approved and registered by MAF as a containment facility
under section 39 of the Biosecurity Act, in accordance with the MAF/ERMA
New Zealand Standard (below), and controls imposed by the Authority (as
follows):
1
Bold headings in the following text refer to Matters to be Addressed by Containment Controls for
Import, Development and Field Testing of Genetically Modified Organisms, specified in the Third
Schedule of the Act.
2
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.
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1.4
The construction and operation of the containment facility shall be in
accordance with:
a) MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.023:
Containment Facilities for Microorganisms.
b) Australian New Zealand Standard AS/NZS 2243.3:20023: Safety in
Laboratories: Part 3: Microbiological Aspects and Containment
Facilities.
c) Physical Containment Level 1 (PC1) requirements of the above
standards for E. coli, P. fluorescens and P. putida and Physical
Containment Level 2 (PC2) requirements of the above Standards for P.
aeruginosa PAO1 and P. syringae pv phaseolicola.
2 To exclude unauthorised people from the facility.
2.1
Construction and operation of the containment facility shall comply with the
requirements of the standards listed in control 1.4 relating to the 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
Construction and operation of the containment facility shall comply with the
requirements of the standards listed in control 1.4 relating to the 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
Construction and operation of the containment facility shall comply with the
requirements of the standards listed in control 1.4 relating to the prevention of
unintended release of the organism by experimenters working with the
organism.
5 To control the effects of any accidental release or escape of an
organism.
5.1
Construction and operation of the containment facility shall comply with the
requirements of the standards listed in control 1.4 relating to controlling the
effects of any accidental release or escape of an organism.
3
Any reference to this standard in these controls refers to any subsequent version approved or endorsed
by ERMA New Zealand
Environmental Risk Management Authority Decision: GMD06004
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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 any viable material 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
standards listed in control 1.4.
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 1.4 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 1.4.
7 Qualifications required of the persons responsible for implementing
those controls.
7.1
The training of personnel working in the facility shall be in compliance with the
standards listed in control 1.4.
_____________________
_____________
Rob Forlong
Date
Chief Executive, ERMA New Zealand
Approval code (BCH code): GMD004162 – 66 (11739 – 43)
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
 Removal of the control regarding inspection of facilities by the Authority, its
agent or enforcement officers
____________________________
Mr Rob Forlong
Chief Executive, ERMA New Zealand
Environmental Risk Management Authority Decision: GMD06004
16 August 2007
Date:
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