Download ENVIRONMENTAL RISK MANAGEMENT AUTHORITY DECISION

ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Amended under s67A on 22 August 2007
26April 2001
Application code
GMC00012
Application type
To import into containment a genetically modified organism under section
40(1)(a) of the Hazardous Substances and New Organisms (HSNO) Act.
Applicant
Date received
Horticulture and Food Research Institute of New Zealand
To import into containment Escherichia coli K12 and B derivatives
containing fragments of DNA that have been cloned from other species.
3 November 2000
Consideration period
4-7 April 2001
Considered by
A Committee of the GMO New Organisms Standing Committee of the
Environmental Risk Management Authority (the Authority).
Purpose
Decision
The application to import genetically modified Escherichia coli into containment is
approved subject to controls in accordance with sections 45(1)(a) and 45(2) of the
Hazardous Substances and New Organism Act 1996.
Organism Identification
The approved organisms are: Escherichia coli strains K12 and B that do not contain
conjugative plasmids or general transducing phages, and that have been genetically modified
by non-conjugative plasmid or bacteriophage vectors.
These vectors shall only contain one or more of the following elements, and involve genetic
modifications that meet Category A or Category B experiments in the Hazardous Substances
and New Organisms (Low-Risk Genetic Modifications) Regulations 1998:
Promoters and terminators
Promoter, operator, enhancer and/or terminator sequences derived from genes sourced from
bacteria, phytoplasma1, insects, molluscs, fungi, elephants or plants, or from bacterial,
fungal or plant viruses.
1
Phytoplasma are bacteria-like parasites that lack cell walls and which cause diseases in plants.
Reporter genes
Fully characterised2 reporter genes whose products can be assayed by one or more of the
following techniques:
 Visual colour or fluorescence
 Spectrophotometrically
 Histochemically
 Enzyme-linked immunosorbent assays (ELISA)
 Thin layer chromatography
 Liquid scintillation counting
 Affinity purification
 Immunological detection
And do not produce proteins that are pathogenic to vertebrates, or vertebrate toxins that have
an LD350 less than 100 μg/kg, or are involved in vertebrate cellular differentiation.
Selectable marker genes
Fully characterised genes that confer the ability to:
1 Tolerate or deactivate antibiotics
2 Tolerate or deactivate metabolic inhibitors
3 Synthesise amino acids
And do not produce proteins that are pathogenic to vertebrates, or vertebrate toxins that have
an LD50 less than 100 μg/kg, or are involved in vertebrate cellular differentiation.
Origins of replication4
Origins of replication derived from Escherichia coli plasmids.
Bacteriophage origins of replication.
Fungal origins of replication.
Other features
1 Multiple cloning site
2 Polyadenylation signals
3 Transcriptional activators, enhancers, responsive elements, receptor elements, and
terminator sequences
4 Secretory signals
5 Compartmental localisation/targeting signals, e.g., nuclear localisation signals
6 Intron sequences to alter gene expression
2
Fully characterised means that the sequence and function of the gene is known.
LD50 is defined as Lethal Dose, 50%. The basic idea (and practice) of the test is to take healthy animals (usually
mice or rats but sometimes dogs, monkeys or other animals) and force feed them enough of a potential toxin to kill
(usually slowly) 50% of them. (Variations include starving the individual before testing, injecting the tested
substance, or coating the animal's skin with the tested chemical.)
3
4
Origins of replication are the nucleotide sequences at which DNA synthesis is initiated.
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Ribosomal binding sites and/or Kozak sequences
Viral packaging signals, e.g., Ψ+, cos sites
Viral long terminal repeat sequences
Viral intergenic regions
Viral genes required for replication
Site specific recombinase systems such as Cre/Lox
Lambda att sites
Specific recognition sites for proteolytic cleavage
Incorporation of genes to allow display of proteins on the surface of filamentous
phage
16 Telomere sequences
17 T-DNA region of Agrobacterium tumefaciens (not including the vir region)
And do not produce proteins that are pathogenic to vertebrates, or vertebrate toxins that have
an LD50 less than 100 μg/kg, or are involved in vertebrate cellular differentiation. Viral
sequences that in toto are capable of producing more than two thirds of a virus shall not be
included in the same E.coli.
Donor Genetic Material
These vectors may contain genetic material sourced from plant viruses, bacteria,
phytoplasma, fungi, plants, invertebrates and elephants (Loxodonta spp. and Elephas spp.),
provided that the donor genetic material shall not come from:
1 New Zealand native or endemic macroflora and macrofauna, or species valued by
Māori that are sourced from New Zealand
2 Species from Appendix 1 of CITES
(http://www.wcmc.org.uk/CITES/eng/index.shtml), unless accompanied by written
approvals from the importing and exporting countries
And that the donor genetic material shall not include:
1 Genes encoding vertebrate toxins that have an LD50 of less than 100 μg/kg
2 More than two thirds of a complete viral genome
3 Sequences that will produce particles able to infect humans.
Application process
The application was formally received on the 3 November 2000, and stalled for further
information, which was received and verified on the 5 March 2001. 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) and
comments from the Department of Conservation.
The application was considered by a Committee of the Genetically Modified Organisms
Standing Committee of the Authority appointed in accordance with section 19(2)(b) of the
HSNO Act 1996. The Committee comprised the following members: Ms Jill White (Chair),
Professor Colin Mantell and Ms Prue Kapua.
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Relevant legislative criteria
The application was lodged pursuant to section 40(1)(a) of the HSNO Act. The decision was
determined in accordance with section 45, taking into account additional matters to be
considered under sections 37 and 44 and matters relevant to the purpose of the Act, as
specified under Part II of the Act.
Consideration of the application followed the relevant provisions of the Hazardous
Substances and New Organisms (Methodology) Order 1998, but with particular regard to
clauses 8 (dealing with the scale and significance of the risks, costs and benefits) and 26
(dealing with applications where the risks are negligible).
Reasons for the decision
Purpose
In accordance with section 45(1)(a)(i) of the HSNO Act 1996, the genetically modified E.
coli strains K12 and B will be used for research purposes as a source of: DNA libraries to
screen for clones, cloned DNA for subcloning into other vectors, cloned DNA for
sequencing, mutagenesis experiments, probes in hybridisation experiments, vectors for DNA
cloning experiments, and for the expression of cloned genes to enable functional analysis of
gene products to assess and gain knowledge of biological processes in humans. The
Committee considers that this constitutes an appropriate purpose under section 39(1)(h) of the
Act 1996: Such other purposes as the Authority thinks fit.
Inseparable organisms
The Committee considered the effects of inseparable organisms in accordance with section
45(a)(ii) of the HSNO Act and noted that the bacterial strains would be imported as pure
cultures, therefore contamination with other organisms is very unlikely. Should any
inseparable organisms be imported, it is very unlikely they would escape containment under
the specified containment controls imposed in this decision.
Ability to escape containment
In accordance with section 44(b) of the HSNO Act the Committee considered the ability of
the organisms to escape from containment.
The Committee requires the genetically modified E. coli to be maintained in a facility
approved under MAF/ERMA New Zealand Standard 154.03.02: Containment Facilities for
Microorganisms. The facility will meet physical containment level 1 (PC1) as described in
the Australia/New Zealand (AS/NZS) Standard 2243.3:1995 Safety in Laboratories: Part 3:
(Microbiology)(this standard is referred to in the rest of the decision as the AS/NZ
Microbiology Standard).
E. coli strains K12 and B, and their derivatives, are classified as Risk Group 1 organisms
according to the AS/NZ Microbiology Standard, which defines Risk Group 1 organisms as:
“(low individual and community risk)- a microorganism that is unlikely to cause
human, plant or animal disease”
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The Committee notes the genetic modification of E. coli meets the characteristics of Category
A and Category B(b)(i) and B(b)(v) experiments under the HSNO (Low-Risk Genetic
Modification) Regulations 1998 if they were developed in New Zealand.
The Committee is satisfied that, taking into account the laboratory procedures proposed by
the applicant and the containment controls imposed in this decision, it is very unlikely that E.
coli could escape from containment in a viable form.
Ability to form self-sustaining populations
In accordance with sections 37(a) and (b) of the Act the Committee also considered the
ability of the organism to establish a self-sustaining population and the ease with which any
such populations could be eradicated.
Should E. coli breach containment the Committee considers that the bacterium is unlikely to
establish a self-sustaining population. In reaching this conclusion, the Committee noted that
the organisms are derivatives of E. coli strains K12 and B, and such strains have been
demonstrated to be less able to survive outside of culture than unmodified bacteria, due to
their requirements for essential amino acids and vitamins (Smith 1975, Heitkamp et al. 1993).
Furthermore, the genetic modifications will not introduce traits to increase environmental
fitness. The Committee notes that the applicant is required to implement procedures for the
retrieval or destruction of any viable material of the organism that has breached containment,
as part of the facility approval, and as detailed in the HortResearch, Mt Albert containment
manual updated October 2000.
The Committee considers that the eradication of the organisms, should escape occur and a
population establish, could be difficult since they may not be able to be detected, and any
eradication method is likely to adversely effect other microorganisms already present in that
environment.
The Committee concludes that should the microorganisms breach containment it is unlikely
to establish self-sustaining populations in the natural environment.
Adverse effects
The Committee considered the potential adverse effects of the organism in accordance with
Section 45(a)(ii) of the HSNO Act.
The types of modifications are restricted to only category A and B experiment as described in
the HSNO (Low Risk Genetic Modification) Regulations, and the vectors will not contain
sequences that are toxic to humans. The Committee concluded that since the host organisms
are very unlikely to survive outside of containment, and that toxins with an LD50 of less than
100μg/Kg are prohibited, it is very unlikely that researchers handling E. coli containing
genetic material from pathogenic organisms will be adversely affected, so long as the
containment controls are adhered to.
The Committee wish to emphasise that this approval does not allow a whole donor organism to
be created from the donor genetic material contained in the E. coli strains. Such an experiment
would involve in vitro manipulation of genetic material which would meet the criteria for a
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genetically modified organism, requiring an approval to develop a new organism. Such a
development is not covered by this approval.
As noted above, the E. coli strains are disabled, requiring specific nutritional supplements,
and are unlikely to survive or establish outside of laboratory culture. Considering the types of
genetic material able to be introduced into these bacteria, the Committee concluded that it is
very unlikely that there will be adverse effects on the environment, so long as the
containment controls are adhered to. The Committee notes that the applicant has many years
experience in handling such organisms without any reported adverse effects.
Based on the fact that the E. coli will be used as a laboratory tool in containment, that
material from native flora and fauna will not be used and other flora and fauna valued by
Māori will not be sourced from New Zealand and that similar microorganisms are already
held in containment the Committee considered that it is very unlikely for there to be adverse
effects on Māori culture.
Negligible risk
Based on the consideration and analysis of adverse effects to the environment and public
health, and taking into account the containment controls imposed in this decision, the
Committee considers risks associated with the importation of E. coli into containment are
negligible. The Committee has therefore considered this application in terms of clause 26 of
the Methodology.
Costs and benefits
Given the application is to undertake research, the Committee is satisfied that the direct
benefits are the increase in scientific knowledge gained by undertaking this research.
The Committee also considered the comments made by Michael Cameron (New Organisms
Officer Department of Conservation), which asked for more information on the benefits of
this application. The Committee considers that the benefits, mainly the scientific knowledge
gained to help understand biological processes, has been sufficiently described.
The Committee is satisfied that costs are unlikely to accrue to parties other than the applicant.
Conclusion
Having considered the possible effects of the organism in accordance with sections
45(1)(a)(ii) and (iii) of the HSNO Act, the Committee is satisfied that the proposed
containment regime and additional controls can adequately contain the organism.
The Committee concludes that, taking account of the ability of the organism to escape from
containment (refer section 44(b) of the HSNO Act 1996), the beneficial effects of having the
organism in containment outweigh the likely adverse effects of the organism and any
inseparable organisms, should the organism escape.
The application to import into containment these genetically modified E. coli strains K12 and
B is thus granted in accordance with section 45(1)(a) of the HSNO Act. As required under
section 45(a)(b) the approval is subject to containment controls, as specified below.
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Containment controls
In order to satisfactorily address the matters detailed in the Third Schedule Part I
Containment Controls for Development, Importations or Field Testing of Genetically
Modified Organisms5 of the Act, the Authority’s approval of this application is subject to the
following controls:
1.
To limit the likelihood of any accidental release of any organism or any viable
genetic material6:
1.1 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 organisms of the Authority’s controls.
1.2 The containment facilities shall be approved by Ministry of Agriculture and Forestry
(MAF) in accordance with the MAF Biosecurity Authority/ERMA New Zealand
Standard 154.03.027 and the controls of the Authority.
1.3 The construction and operation of the containment facilities (‘the facility’) in which the
organisms are maintained, shall be in accordance with the:
a)
MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.027: Containment
Facilities for Micro-organisms, and
b)
Australian New Zealand Standard AS/NZS 2243.3:19957 Safety in Laboratories: Part 3:
(Microbiology), at Physical Containment Level 1 (PC1).
2.
To exclude unauthorised people from the facility:
2.1 The identification of entrances, numbers of and access to entrances, and security
requirements for the entrances and the facility shall be in compliance with the
requirements of the standards listed in control 1.3.
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 1.3.
Bold headings refer to Matters to be Addressed by Containment Controls for Development and Field Testing of Genetically
Modified Organisms, specified in the Third Schedule 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, eg
when organisms or parts thereof are sublethally 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
7
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4.
To prevent unintended release of the organism by experimenters working with
the organism:
4.1 The prevention of unintended release of the organism by experimenters working with the
organism shall be in compliance with the standards listed in control 1.3.
5.
To control the effects of any accidental release or escape of an organism:
5.1 Control of the effects of any accidental release or escape of an organism shall be in
compliance with the standards listed in control 1.3.
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 the contingency plan for the attempted
retrieval or destruction of any viable material of the organisms that have escaped shall be
implemented immediately. The contingency plan shall be included in the containment
manual in accordance with MAF Biosecurity Authority/ERMA New Zealand Standard
154.03.027: Containment Facilities for Micro-organisms.
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 control 1.3.
6.2 The containment manuals shall be updated, as necessary, to address the implementation
of the controls imposed by this approval, in accordance with MAF Biosecurity
Authority/ERMA New Zealand Standard 154.03.027: Containment Facilities for Microorganisms.
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 1.3.
_____________________________
Date: 26 April 2001
Ms Jill White Chair,
Genetically Modified Organisms
Standing Committee of the Authority
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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
 Removal of the control regarding inspection of facilities by the Authority, its agent or
enforcement officers
____________________________
Dr Kieran Elborough
Chair, GMO Standing Committee
Date: 22 August 2007
References
Heitkamp MA, et al. (1993). Fate in sewage of a recombinant Escherichia coli K-12 strain
used in the commercial production of bovine somatotropin. J. Ind. Microbiol. 11, 243-252.
Smith HW (1975). Survival of orally administered E. coli K12 in alimentary tract of man.
Nature 255, 500-502.
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