ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Amended under s67A on 23 August 2007
8 April 2003
Application code
GMC02011
Application type
To import into containment organisms under section 40(1) (a) of the
Hazardous Substances and New Organisms (HSNO) Act.
Applicant
University of Otago, Dunedin
Purpose
To import into containment genetically modified Lactobacilli reuteri strain
100-23 derived by insertion of a replication –defective plasmid to study the
properties essential for colonisation of the murine gut, in order to better
understand the microbial ecology of the gastrointestinal tract.
Date received
28 January 2003
Consideration date
18 March 2003
Considered by
A Committee of the Genetically Modified Organisms Standing
Committee of the Environmental Risk Management Authority (the
Authority).
1 Summary of Decision
The application to import into containment genetically modified Lactobacilli reuteri, is approved
with controls, having been considered in accordance with the relevant provisions of the Hazardous
Substances and New Organisms (HSNO) Act 1996 and the HSNO (Methodology) Order 1998.
The organisms have been given the following unique identifiers for the ERMA New Zealand
Organism Register:
Lactobacillus reuteri strain 100-23 insertional mutants derived by the integration of the
replication-defective vector pOR128
The scope of the organisms subject to the approval is limited to Lactobacillus reuteri 100-23
insertional mutants created by the integration of the replication-defective vector pOR128 that have
been modified with genetic material which may encode properties essential for the colonisation of the
murine gut. No DNA sequences other than those derived from the vector pOR128 and/or L. reuteri
shall be inserted into the transformed L. reuteri during the creation of the mutants. The site of insertion
is not defined and individual L. reuteri imported may vary in both their point of insertion and the
phenotypic consequences of the insertion.
The modified L. reuteri will be imported into a laboratory registered under MAF/ERMA New Zealand
standard 154.03.02: Containment facilities for micro-organisms at Physical Containment Level 2
(PC2) as defined in AS/NZS 2243.3.2002. Safety in Laboratories. Part 3: Microbiological aspects and
containment facilities.
2 Legislative Criteria for the application
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 section 44, and matters relevant to the purpose of the Act, as specified under Part II of the
HSNO Act. Unless otherwise stated, references to section numbers in this decision refer to sections of
the HSNO Act.
Consideration of the application followed the relevant provisions of the Hazardous Substances and
New Organisms (Methodology) Order 1998 (the Methodology), with particular regard to clauses 12
(dealing with assessment of risks) and clauses 13 (dealing with assessment of costs and benefits).
Unless otherwise stated, references to clause numbers in this decision refer to clauses of the
Methodology.
3 Application Process
3.1 Application receipt
The application was formally received by ERMA New Zealand on 28 January 2003 and verified as
containing sufficient information on 28 January 2003.
The organisms to be imported are defined as low risk (category B (b) (IV) (A)) as defined in the
HSNO (Low-Risk Genetic Modification) Regulations 1998, and therefore the application was not
publicly notified according to ERMA New Zealand policy.
In accordance with clauses 2(2) (e) and 5 of the Methodology and section 58(c) of the HSNO Act the
Department of Conservation (DoC) and Ministry of Agriculture and Forestry (MAF) were invited to
comment on this application.
3.2 Information available for consideration
The documents available for the consideration of this application by the Committee included the
application and its appendices; the University of Otago Containment Manual for the Vertebrate
Laboratory Animals facility; the Containment and Quarantine Manual for holding of Microorganisms; and an Evaluation and Review (E&R) Report prepared by ERMA New Zealand. Neither
the Department of Conservation nor the MAF Biosecurity Authority had any specific comments on
this application.
3.3 Decision making Committee
The application was considered by the Genetically Modified Organism Standing Committee of the
Authority, appointed in accordance with section 19(2) (b) of the HSNO Act. The Committee
comprised the following members: Ms Jane Lancaster (Chair) and Dr Lin Roberts.
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4 Consideration
4.1 Purpose of the Application
The purpose of the application is to import into containment genetically modified Lactobacilli reuteri
that contain a replication defective plasmid incorporated into their genome which alters their ability to
colonise the murine gut. These organisms will be tested for their colonisation ability using a unique
colony of Lactobacillus-free mice which are maintained within the Department of Microbiology,
University of Otago.
In accordance with section 45(1) (a) (i) of the HSNO Act, the Committee determined that the purpose
was appropriate under section 39(1) (h): Such other purposes as the Authority thinks fit.
4.2 The Sequence of Steps in the Consideration
In accordance with clause 24 of the Methodology, the approach adopted by the Committee was to look
sequentially at identification, assessment and evaluation of risk, costs and benefits. Interposed with
this was the consideration of the adequacy of the proposed containment regime, and the ability for the
organisms to escape and to form self-sustaining populations. Management techniques were considered
in relation to the identified risks (clause 24) and those risks identified as significant were assessed
(clause 12). Costs and benefits were assessed in accordance with clause 13 of the Methodology.
Risk characteristics were then established, in accordance with clause 33 of the Methodology.
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, and the cost-effectiveness of the application of controls was considered in accordance with
clause 35.
4.3 Organism description
The scope of the organisms subject to the approval is limited to Lactobacillus reuteri 100-23
insertional mutants created by the integration of the replication-defective vector pOR128 that have
been modified with genetic material which may encode properties essential for the colonisation of the
murine gut. No DNA sequences other than those derived from the vector pOR128 and/or L. reuteri
shall be inserted into the transformed L. reuteri during the creation of the mutants. The site of insertion
is not defined and individual L. reuteri imported may vary in both their point of insertion and the
phenotypic consequences of the insertion.
4.4 Identification of the potential significant risks, costs and benefits
No significant risks or costs were identified for assessment and evaluation in terms of clauses 9(a),
9(b)(i), 9(c)(i), 9(c)(ii), 10(a), 10(b) and 10(d)-(f) of the Methodology, which incorporates sections 5,
6, and 8 of the Act. In reaching this conclusion, the Committee gave particular regard to:
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Risks to the environment
The Committee considered there were no significant adverse effects on the environment associated
with the import into containment of genetically modified Lactobacilli reuteri. The Committee noted
that the wild-type strain, Lactobacillus reuteri 100-23, was first isolated in New Zealand, is highly
rodent specific, and is very unlikely to persist in the gut of other animals. Because the site of insertion
of the replication defective plasmid and its effect on the colonisation ability of L reuteri is unknown,
there is some uncertainty associated with the characteristics, and hence the risk level of those
characteristics, of the mutant strains. Identification of nature of the mutants at importation (Control
1.7) will provide a greater level of certainty about the characteristics and thus about the risk level of
the mutant strains. Control 1.8 requires the applicant to notify ERMANZ of any mutant strains with
pathogenicity.
However, the Committee considered that as the long experience with L. reuteri has not
revealed any mutants with pathogenicity and as the modified strains are very unlikely to persist in
competition with unmodified Lactobacillus reuteri, the level of risk to the environment was not
significant.
Risks to public health
The Committee considered there were no significant adverse effects on human health associated with
the genetically modified Lactobacilli reuteri. The Committee noted that Lactobacilli species are nonpathogenic and are accorded GRAS (Generally Regarded As Safe) status by the United States Federal
Drug Administration.
Risks to Māori and their taonga
The Committee noted that the organisms will be held in containment and the modifications do not
involve genetic material from native or valued species or from humans. The Committee therefore
considers that there are no significant adverse effects on the relationship between Māori culture and
their traditions with their ancestral lands, water, sites, waahi tapu, valued flora and fauna and other
taonga, associated with this application.
Costs and benefits (beneficial effects)
The Committee addressed potential costs and benefits associated with the application, in accordance
with the Methodology clauses 9, 10, 13, and 14 and section 6(e) of the HSNO Act. The Committee
considered that no significant costs were associated with the import of genetically modified L. reuteri
into containment. The Committee identified the primary beneficial effects as those related to increased
scientific knowledge associated with the research
As the Committee did not identify any potential significant risks (and associated costs) associated with
the import into containment of genetically modified L. reuteri, no further consideration and/or
assessment of these aspects is warranted.
4.4 Adequacy of the proposed containment
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
at the same time in an integrated fashion. This is because the former interact with the latter and this is
recognised in clause 12(d) of the Methodology and in section 45(1) (a) (ii) of the Act. For
convenience in setting out the decision the adequacy of containment is discussed first.
Ability to escape from containment
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The Committee considered that if the genetically modified organisms intended to be imported were
developed in New Zealand; they would meet the characteristics of Category B experiments
(specifically B (b) (IV) (A)) under the HSNO (Low-Risk Genetic Modification) Regulations 1998.
Experiments involving these organisms are thus required to be conducted within a facility registered in
accordance with MAF/ERMA New Zealand standard 154.03.02: Containment facilities for microorganisms, at Physical Containment Level 2 (PC2) as specified in the Australian/New Zealand
Standard AS/NZS 2243.3:2002. Safety in Laboratories: Part 3: Microbiological Aspects and
Containment Facilities (refer to Controls 1.2 and 1.3 in Annex 1 of this decision).
The Committee noted that Lactobacilli do not produce spores, so are likely to be effectively contained
without additional controls on their handling.
The Committee noted that the applicant intends to use mice to investigate the colonisation ability of
the genetically modified Lactobacilli reuteri. Therefore controls are also imposed within this decision
with regard to mice that may contain the genetically modified Lactobacilli. These controls require that
the mice used are to be housed in a facility registered under both MAF/ERMA New Zealand standard
154.03.02: Containment facilities for micro-organisms, and MAF/ERMA New Zealand standard
154.03.03: Containment Facilities for Vertebrate Laboratory Animals, at Physical Containment Level
2 (PC2) as defined in the Australian/New Zealand Standard AS/NZS 2243.3:2002: Safety in
Laboratories: Part 3 (Microbiology).
Experiments involving mice will be carried out within gnotobiotic isolators1. The Committee noted
that the treatment and disposal of waste products and bedding is covered in section 5.10 of the
University of Otago, Dunedin Campus: Vertebrate Laboratory Animals Containment Manual. The
Committee has elected to impose an additional control (control 1.5) which requires that waste material
generated within the vertebrate facility (including bedding) shall be sterilised by autoclaving. The
carcasses of mice used in the experiments shall be carried in a double-bag to a biological safety
cabinet within the PC2 laboratory. Following the removal of organs, carcasses shall be returned to the
bag and disposed of by incineration. Remains of organs shall also be disposed of by incineration. The
interior of the gnotobiotic isolator shall be sprayed with sterilising solution prior to it being opened for
cleaning.
The Committee considers that with the containment controls it has imposed (refer to Annex 1 of this
decision); it is very unlikely for viable genetically modified cell lines to escape or be removed
inadvertently from containment.
Ability of the organism to establish a self-sustaining population outside of containment and the
ease of eradication
In accordance with sections 44 and 37 of the HSNO Act the Authority considered the ability of the
organisms to establish undesirable self-sustaining populations, should they escape from containment,
and the ease with which such populations could be eradicated. In evaluating these matters the
Committee took into account the nature of the organisms.
The Committee noted that the genetically modified Lactobacillus reuteri will have an undetermined
ability to colonise the murine gut but the majority of strains will be defective in comparison to the wild
type. As a consequence the mutants are unlikely to survive competition with un-modified
Lactobacillus reuteri. The Committee also noted that (based on information provided by the applicant)
the parent strain of Lactobacillus reuteri used to produce the genetically modified strains of
Lactobacillus reuteri to be imported are very unlikely to persist in the gut of humans or other animals,
should they escape from containment.
A gnotobiotic isolator is a container that houses or isolates an animal from unwanted association or infection with
micro- and macro-organisms
1
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The Committee concurs with the ERMA New Zealand E&R report (section 4.5.5), that if the modified
L. reuteri escaped from containment they are very unlikely to be able to survive and establish selfsustaining populations. Furthermore, the Committee is satisfied that the controls imposed within this
decision ensure that inadvertent loss of material from containment is very unlikely. Given that the
organisms would be unlikely to survive outside of containment and that in the unlikely event that the
organism escaped from containment, the Committee concluded that eradication of the organisms
would not be necessary.
4.5 Assessment of benefits (beneficial effects)
A benefit is defined in clause 2 of the Methodology Order as “the value of a particular positive effect
expressed in monetary or non-monetary terms”. Benefits that may arise from any of the matters set out
in clauses 9 and 10 of the Methodology were considered in terms of clause 13.
Under clause 13(a), the Committee considered the primary benefit associated with this application to
be increased scientific knowledge (non-monetary) gained through studying the colonisation properties
of Lactobacillus reuteri in order to better understand the microbial ecology of the gastrointestinal tract.
The Committee noted that while the magnitude and expected value of this benefit (clause 13(b)) is
uncertain, immediate benefits are likely to accrue directly to the applicant (clause 13(c)).
4.6 Overall evaluation of risks, costs and benefits
The overall evaluation of risks, costs and benefits set out below was carried out having regard to
Clauses 22 and 34 of the Methodology and in accordance with the tests in clause 26 of the
Methodology and section 45 of the Act.
Clause 34 of the Methodology sets out the approaches available to the Authority in evaluating the
combined impact of risks costs and benefits i.e. weighing up risks, costs and benefits.
The Committee is satisfied that the modified L. reuteri can be adequately contained (sections 45(1) (a)
(iii) and 44(b) of the HSNO Act) under the controls required by this decision (refer to Annex 1). In
relation to the additional matters to be considered under section 37 of the HSNO Act, the Committee
considered that it is very unlikely for genetically modified L. reuteri to escape or be removed
inadvertently from containment and form a self-sustaining population, and that eradication would be
unnecessary given that the organisms would be unlikely to survive outside of containment.
The Committee did not identify any potential significant risks to the environment, to public health and
safety, or to Māori and their taonga associated with this application.
The Committee therefore considered this application in terms of clause 26 of the Methodology, and
thus considered that the benefits associated with the importation and use of the modified L. reuteri in
containment outweighed the costs. In reaching this conclusion, the Committee considered all the
possible beneficial and adverse effects of the organisms in accordance with sections 45(1) (a) (ii) and
(iii) of the HSNO Act.
The Committee considered that it is inappropriate to impose a restricted time limit on this approval
because the approval is likely to be used over a period of time. The Committee also noted that the
controls imposed within this approval have been used in previous approvals, and as such do not
consider that it is necessary for the applicant to report to ERMA New Zealand on the exercise of the
controls imposed by this decision.
In addition, the Committee considered that additional controls to measure or monitor for adverse
effects are not necessary in this case since the work will be conducted within containment facilities.
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5 Decision
Pursuant to section 45(1) (a) (i) of the Act, the Committee is satisfied that this application is for one of
the purposes specified in section 39(1) of the Act, being section 39(1) (h): Such other purposes the
Authority sees fit.
Having considered all the possible effects of the organisms in accordance with sections 45(1)(a)(ii)
and (iii) of the Act 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 this decision, the view of the Committee is that the risks (or costs) of adverse
effects associated with the importation into containment of the organism(s) are outweighed by the
benefits of conducting the research.
The Committee is satisfied that the proposed containment regime together with the additional control
imposed will adequately contain the organisms as required by section 45(1)(a)(iii) of the Act.
In accordance with clause 36(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 following sections in the Act - section 5(a), section
5(b), section 45(1)(iii), and the following clauses in the Methodology: clauses (9), (10), (12), (13),
(21), (22), (24), (25), (26), (34).
The application for importation into containment of genetically modified Lactobacillus reuteri is thus
approved, in accordance with section 45(a) of the HSNO Act. As required under section 45(2) the
approval is subject to controls (as listed in Annex 1 of this decision).
________________________
Date: 8 April 2003
Ms Jane Lancaster
Genetically Modified Organisms
Standing Committee of the Authority
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: 23 August 2007
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Annex 1: Controls required by this approval
In order to satisfactorily address the matters detailed in the Third Schedule Part I:
Containment controls for importing, developing or field testing of genetically modified
organisms2 of the HSNO Act, and other matters in order to give effect to the purpose of the
HSNO Act (section 45(2)), 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
material3:
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.
The containment facility in which the genetically modified Lactobacillus reuteri are maintained
shall be registered by the Ministry of Agriculture and Forestry (MAF) Biosecurity Authority in
accordance with:
 MAF/ERMA New Zealand Standard 154.03.024: Containment Facilities for Microorganisms, at Physical Containment Level 2 (PC2) as defined in AS/NZS 2243.3.20024.
Safety in Laboratories. Part 3: Microbiological aspects and containment facilities.
The containment facility in which mice, used to investigate the colonisation ability of the
genetically modified Lactobacillus reuteri, are maintained shall be registered by the Ministry of
Agriculture and Forestry in accordance with:
 MAF/ERMA New Zealand standard 154.03.024: Containment Facilities for Microorganisms.
 MAF/ERMA New Zealand standard 154.03.034: Containment Facilities for Vertebrate
Laboratory Animals.
At Physical Containment Level 2 (PC2) as defined in AS/NZS 2243.3.20024. Safety in
Laboratories. Part 3: Microbiological aspects and containment facilities.
The construction and operation of the containment facilities (‘the facility’) in which the organisms
are maintained, shall be in accordance with the relevant standards listed in 1.2 and 1.3 above.
Waste material generated within the vertebrate facility (including bedding) shall be bagged within
gnotobiotic isolators, and sterilised by autoclaving. The carcasses of mice used in the experiments
shall be carried in a double-bag to a biological safety cabinet within the PC2 laboratory. Following the
removal of organs, carcasses shall be returned to the bag and disposed of by incineration. Remains of
organs no longer required shall also be disposed of by incineration. The interior of the isolator shall be
sprayed with an appropriate sterilising solution prior to it being opened for cleaning.
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.
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, eg
when organisms or parts thereof are sublethally damaged by being frozen, dried, heated, or affected by chemical.
3
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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The Lactobacillus reuteri 100-23 insertional mutants must be imported from a recognised
laboratory that is involved in research on Lactobacilli. Individual mutants must be derived from pure
cultures that contain no inseparables.
The applicant shall report to ERMA NZ on the identity of each mutant as it is imported. Details of
the genetic material of each construct (including recognised laboratory source, DNA sequence, site of
insertion and potential gene products and function) shall be provided to ERMA NZ.
Any imported strains of Lactobacillus reuteri, which are subject to this approval, that
exhibit pathogenicity shall be reported to ERMA NZ immediately.
2.
To exclude unauthorised people from the facility:
Construction and operation of the containment facility shall comply with the requirements of the
standards listed in controls 1.2 and 1.3 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:
Construction and operation of the containment facility shall comply with the requirements of the
standards listed in controls 1.2 and 1.3 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:
Construction and operation of the containment facility shall comply with the requirements of the
standards listed in controls 1.2 and 1.3 relating to the prevention of unintended release of the
organisms by experimenters working with the organisms.
5.
To control the effects of any accidental release or escape of an organism:
Construction and operation of the containment facility shall comply with the requirements of the
standards listed in controls 1.2 and 1.3 relating to controlling the effects of any accidental release or
escape of an organism.
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.
In the event of any breach of containment of the organisms, 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 controls 1.2 and 1.3.
6.
Inspection and monitoring requirements for containment facilities:
The operation of the containment facilities shall comply with the requirements contained in the
standards listed in controls 1.2 and 1.3 relating to the inspection and monitoring requirements for
containment facilities.
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The containment manuals shall be updated, as necessary, to address the implementation of the
controls imposed by this approval, in accordance with the MAF /ERMA New Zealand Standard
154.03.024: Containment Facilities for Micro-organisms.
7.
Qualifications required of the persons responsible for implementing those controls:
The training of personnel working in the facility shall be in compliance with the standards listed in
controls 1.2 and 1.3.
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