ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Controls amended under S67A 27 March 2002.
Application code
Hearing date
Considered by
GMF99004
Original Decision
26 October 2000
6 and 7 April 2000
Special Committee of the Authority appointed under section 19(2)(b) of
the Hazardous Substances and New Organisms Act 1996.
Application Details
Application code
Applicant
Purpose
Date application received
ERMA New Zealand contact
GMF99004
New Zealand Pastoral Agricultural Research Institute Ltd (AgResearch)
To field test in containment in the Waikato region, genetically modified sheep
with an inactivated myostatin gene, to increase the understanding of
myostatin function in order to identify the effects on sheep muscularity
16 June 1999
Erika Anderson
Decision
The application is approved with controls for a period of five years from the date of this
decision.
The organism is:
Ovis aries (sheep)
Construct: Plasmid, pBlueScript, containing the sheep myostatin locus (comprising three exons
and two introns), a copy of a neomycin (neo) or puromycin (antibiotic resistance) gene originally
derived from the bacteria Escherichia coli (E. coli), and a thymidine kinase (tk) gene. Refer to Annex
1 for further details of the genetic construct.
Phenotype: ‘myostatin knockout’ sheep (disruption of the ovine myostatin gene by introduction
of the neomycin or puromycin resistance gene into the first exon), resulting in sheep that lack a
functional myostatin protein.
Application Process
The application was formally received on 16 June 1999 and verified on 3 December 1999,
following a number of additional information requests.
The application was publicly notified on 8 December in The Dominion, The New Zealand Herald,
The Press and The Otago Daily Times.
Page 1 of 36
Public submissions closed on 16 February 2000. Eighty submissions were received, of which
sixty-four submitters requested to be heard at a public hearing in support of their submission. A
list of submitters is attached as Annex 2.
The documents available for the evaluation and review of the application by ERMA New
Zealand included: the application and appendices (including supporting documentation and
confidential information provided), public submissions, and submissions and comment from
other government agencies (including the Ministry of Agriculture and Forestry [MAF], the
Department of Conservation [DOC], and the Ministry of Health).
In accordance with section 19(2)(b) of the Hazardous Substances and New Organisms (HSNO)
Act 1996, the Authority appointed a Committee to determine the application. The Committee
comprised members from the Authority, including: Bill Falconer (Chair), Professor Barry Scott,
Dr Oliver Sutherland, Professor Colin Mantell, Dr Lindie Nelson, and one external member, Dr
Mere Roberts appointed for her expertise and knowledge in Māori culture and traditions).
Hearing
A public hearing was held on 6 and 7 April 2000 in Wellington.
Presentations
Presentations were made to the Committee by the following parties:
For the applicant:
1.
2.
3.
4.
Michael Holm
Dr John Bass
Dr Ravi Kambadur
Jim Napier
Partner, Russell McVeagh (Legal Counsel)
Programme Leader, Functional Muscle Genomics (AgResearch)
Senior Scientist, Functional Muscle Genomics (AgResearch)
Research Associate, Functional Muscle Genomics (AgResearch)
For ERMA New Zealand:
1.
Elizabeth Beale
Project Leader, ERMA New Zealand
For Ngā Kaihautū Tikanga Taiaoi:
1.
Gerrard Albert
Ngā Kaihautū Tikanga Taiao
Submitters:
1.
2.
Sue Kedgley
Tom Bennion
3.
4.
Wendy McGuinness
Chris Lester (sheep &
Beef Farmer)
Mark Christensen
(Anderson Lloyd)
William Rolleston
Sigi Kirchmair
5.
6.
7.
Green Party
Legal Counsel representing Sue Kedgley and Associate
Professor Peter Wills
Private
Representing Federated Farmers of New Zealand (Inc)
Legal Counsel representing New Zealand Life Sciences
Network and Biotenz
Representing New Zealand Life Sciences Network and Biotenz
Private
Page 2 of 36
8.
9.
10.
11.
12.
13.
Kei Munro
Briana Ball
Susie Lees
Claire Bleakley
Noel K Wierzbicki
Manfred von
Tippelskirch
14. Tscherner Wolfgang
15. Mary Anne HowardClarke
16. Bob McDougall
17.
18.
19.
20.
21.
Trudy Burgess
Alex Vickers
Joanna M Paul
Tracy Buick
Rich J Wernham
Private
On behalf of Berylla Berylla
Private
Private
Private
Private
Private
GE Free New Zealand in Food and Environment (RAGE Inc)
Witness to Mary Anne Howard-Clarke [GE Free New Zealand
in Food and Environment (RAGE Inc)]
GE Free NZ
Private
Private
Private
Private
Further information
Additional information sought from the applicant and considered by the Committee included:
1
Detailed information in respect of the outdoor containment facility, including both the
currently registered 4 ha facility, and the proposed expanded 45 ha facility to enable the
Committee to consider and form a view as to the probability of the escape of sheep by
theft (sabotage) from the containment facilities through negligence, or failure in systems
and processes in place.
2
Further detailed information on the electronic subcutaneous microchips proposed to be
inserted into genetically modified sheep:
i.
the detailed specifications of the proposed microchips
ii.
information on their capability to facilitate detection and retrieval of any sheep that
may escape from the containment facilities.
3 Further information on animal ethics approvals, including:
i.
copies of the full applications and decisions of the Ruakura Animal Ethics
Committee (AEC) in respect of work covered under this application
ii.
a list of the members, and their respective qualifications, of the Ruakura AEC
Following receipt and consideration of the information specified above, the Committee sought
further additional information, including:
i
the PGSF Report relating to the funding of the Contract of which the proposed research
forms part;
Page 3 of 36
ii
further commentary from the Ruakura Animal Ethics Committee on certain aspects of
the monitoring of the proposed research.
The information received largely provided further elaboration and detail on material already
available, and was not considered by the Committee to constitute new information. In addition
sections of this information were provided in confidence to the Committee. However, the
following were forwarded to parties to the application:
i
Extracts from Section 1 – Parliamentary Section - the PGSF Report (1999-2000)
ii
Letter from Applicant to ERMA New Zealand dated 14 August containing
information from the Ruakura Animal Ethics Committee
iii
Letters from the Chairman of the Ruakura Animal Ethics Committee to the
Applicant dated 27 July and 15 August 2000.
Relevant Legislative Criteria
The application was lodged pursuant to section 40 of the Hazardous Substances and New
Organisms Act 1996, and determined in accordance with section 45, the additional matters
contained in sections 37 and 44, and the matters set out in Part II of the Act.
Consideration of the application followed the relevant provisions of the Hazardous Substances
and New Organisms (Methodology) Order 1998 (the Methodology).
The Application
The application is for approval to produce and field test in containment, Ovis aries (sheep)
containing a genetic modification aimed at disrupting the ovine myostatin gene locus. The
modification involves the introduction of a construct containing an ovine myostatin locus
disrupted by the introduction of a neomycin (neo) or puromycin (antibiotic) resistance marker
gene. The construct also contains a thymidine kinase (tk) marker gene.
Sheep that are homozygous for the modification are expected to lack the ability to express
functional myostatin protein. Myostatin is a protein that is believed to restrict muscle growth and
development. The purpose of the work is to increase the understanding of myostatin function in
order to identify the effects on sheep muscularity.
This application seeks approval to produce and field test myostatin knockout sheep, from
embryos developed under delegated approval. The development of myostatin knockout sheep
embryos was approved by the Ruakura Institutional Biological Safety Committee (IBSC) under
delegated authority from the Environmental Risk Management Authority (the Authority) on 17
March 1999 (Application code: GMO99/ARR001, ERMA approval code: GMD000056).
The field test will comprise of up to 100 sheep divided into three flocks of approximately 30
genetically modified sheep each.
Page 4 of 36
The genetically modified sheep will be maintained in containment at the Ruakura Research
Centre (RRC), in, at various stages, an indoor or outdoor containment facility1.
The indoor facility is a registered containment facility under the Biosecurity Act 1993, in
accordance with the Ministry of Agriculture and Forestry (MAF) Biosecurity Authority/ERMA
New Zealand Standard Animal Health and Welfare Standard 154.03.03: Containment Standard for
Vertebrate Laboratory Animals.
The outdoor facility (45ha site) is registered as a containment facility in accordance with the
MAF Biosecurity Authority/ERMA New Zealand Animal Health and Welfare Standard
154.03.06: Containment Standard for Field Testing of Farm Animals.
Scope of the application
The development of genetically modified embryos has been approved under the HSNO Act
1996 by AgResearch’s Ruakura IBSC under delegated authority from the Environmental Risk
Management Authority (the Authority).
The application identifies options for marker sequences that may be utilised in constructs to
produce sheep of the phenotype identified. The controls on this approval specify that, any
further constructs and genetically modified embryos to be developed, using alternate marker
sequences, must be approved by AgResearch Institute’s Ruakura IBSC prior to production and
field testing of genetically modified sheep. Schedule 1 to this decision defines the scope of any
further constructs to be utilised for the production and field testing of sheep under this approval.
The total number of sheep involved in the field test, including genetically modified and
conventional sheep, will not exceed the capacity of the containment facilities, as approved under
the MAF Biosecurity Authority/ERMA New Zealand Standards, nor any requirements of the
Ruakura AEC, and shall be the minimum number of animals required to obtain statistically
significant results.
In essence, this application covers an extension of research already undertaken on the
development of the genetically modified sheep embryos. The flock is to be maintained in
containment, and there is no intention for the meat, milk or offal from the sheep to enter the
human food chain.
Jurisdiction to consider the application as a field test in
containment
A submitter questioned whether the application was properly lodged as a “field test in
containment” and therefore whether the Committee had jurisdiction to hear it.
The applicant sought approval to field test the sheep to be developed on the advice of ERMA
New Zealand, on the basis that the purpose of the research is to increase understanding of
myostatin function in order to identify the effects on sheep muscularity, during the life-cycle of
the sheep, and under normal conditions ‘in the field’. The applicant might equally have sought
approval to develop the sheep as genetically modified organisms. The only practical difference
A containment facility refers to a place that is registered by MAF under the Biosecurity Act 1993 as a containment
facility in accordance with approved standards.
1
Page 5 of 36
between an application to field test and one to develop is that as regards the latter, the Authority
has a discretion whether or not to notify the application, whereas for a field test the Act requires
that the application be publicly notified, and submissions on it invited. Where the research
proposed in an application is strictly neither a development nor a field test (in accordance with
the definitions in the HSNO Act 1996), then in advising the applicant, the key consideration for
ERMA New Zealand is the nature of the environment in which the research will be undertaken.
As the intention is to undertake a portion of the research ‘in the field’, the applicant was advised
to make an application to field test in containment rather than to develop the genetically modified
sheep. Both the applicant and the Authority accepted this view.
The distinction between a development in containment and a field test in containment is, in this instance,
one of presentation rather than substance, and the Authority is of the view that it has the
jurisdiction to consider the application as a field test, and that this jurisdiction is not
compromised by the fact that the application might also have been considered as a development in
containment.
Purpose of the application
At the hearing a number of submitters expressed concern at what they perceived to be a change
in emphasis with respect to the purpose and potential benefits of the proposed research
programme. Specifically, they discerned from the application, that a principal objective behind
the stated purpose of increasing understanding of the myostatin function in order to identify the
effects on sheep muscularity, was to enable the applicant to identify the scope to develop a
variety of heavy muscled sheep in New Zealand for agricultural benefit. In the course of the
hearing however, the applicant presented benefits from potential medical applications of the
information as being of as equal if not greater importance than these from potential agricultural
applications.
Submitters argued that this change in emphasis put them at a disadvantage; some saying that they
may not have made submissions at all had they believed the application was aimed at potential
medical applications. At the risk of over generalising, the submissions seemed to accept that
some account might be taken of potential medical benefits, where these had a high degree of
certainty as to their outcome, but did not accept that account should be taken of potential
benefits in any other area.
These submissions required the Committee to consider two related issues in relation to
applications whose immediate purpose is obtaining scientific information. First, the question of
the extent to which the Committee should take into account benefits which may flow from
future developments involving that information; and secondly whether the Committee could set
any thresholds in relation to the value or benefit of the scientific information being sought.
The issue of how potential future benefits should be weighed by the Authority has been
addressed in previous applications. The Committee has taken the view to date, that the principal
benefit of research programmes such as that involved in the present application is the scientific
knowledge they will generate, and that undefined potential future benefits, be they of a medical,
agricultural or economic nature, should not be taken into account by the Committee in its
assessment of benefits. Any future risks, costs or benefits that may result from the release of an
organism, or its commercial use as a food product, or in a pharmaceutical, agricultural or medical
context, are more appropriately dealt with in the context of any future applications that may be
made for those purposes.
Page 6 of 36
As a general matter, the Committee was of the view that where, as in the present instance, the
research programme had been considered and given appropriate funding priority by the
applicant, and approved by the Foundation for Research Science and Technology, the
Committee might assume, in the absence of evidence to the contrary, that the scientific
knowledge to be gained was beneficial in itself, regardless of whatever future benefits may
emerge from its use. The sections of the Public Good Science Fund Report to Parliament 19992000 relating to the proposed research endorsed its value.
Overall, the Committee concluded, on the basis of the application and the PGSF Report, that
information on sheep muscularity would be beneficial in itself, not least because it would likely
contribute to a wide range of related research. The Committee did not consider it could more
precisely establish immediate benefit thresholds for research of this nature, for example by
attaching greater benefit to information which may lead to medical outcomes than that which
may lead to agricultural outcomes. Such an approach may encourage applicant’s to misrepresent
potential benefits. Nor was the Committee prepared to conclude that this was an area of inquiry
which should not be pursued, even though the ultimate applications of the research and the
benefits which might flow from it were uncertain.
The Committee accepted that the information could only be obtained efficiently through
knockout procedures, noting the applicant’s statement that the heavy muscled sheep variety in
New Zealand, Texel, and the European Bextel, are not known to have a mutation for myostatin,
and would therefore not be appropriate for this research.
The Committee noted that while one of the research outcomes may be the development of
heavy muscled sheep, which might alternatively be pursued through conventional breeding
techniques, the applicant’s objective as stated to the PGSF was physiological studies associated
with the development of the sheep rather than the sheep themselves. The applicant pointed out
that the programme is not looking at muscularity per se, but is determining the function of the
myostatin gene using sheep as an animal model.
The Committee accepted that though different ethical issues may arise (dealt with subsequently
in this decision) there were benefits to be gained from research using larger animals such as
sheep (in which the applicant has considerable scientific expertise), rather than smaller laboratory
animals.
Key Issues
The Committee’s consideration of the application encompassed those matters relevant to the
application, and included:
1.
Adequacy of the proposed containment regime, including:
i.
the ability of the organism or any heritable material to escape from containment,
including:
(a)
escape of whole sheep:

escape by human intervention, including as a result of:
-

sabotage (including theft)
negligence
events of force majeure (including natural disaster)
Page 7 of 36
(b)
(c)
2.
escape of genetic material by horizontal gene transfer, via:

biting arthropods

microorganisms
disposal of sheep and other biological material
ii.
the ability of the organism to establish a self-sustaining population
iii.
the ease of eradication of any population established.
risks associated with the organism, including:
i.
risks to public health from the consumption of meat, milk or offal derived from
genetically modified sheep, in the event that sheep enter the national flock, and
subsequently, the food chain, following an escape from containment, including:
(a)
direct health effects as a result of consumption
(b)
development of antibiotic resistance as a result of horizontal gene transfer to
gut bacteria
(c)
unknown outcomes, ie CJD and long-term unanticipated health effects
ii.
risks to New Zealand’s ‘clean green image’, and export relationships and organic
farming
iii.
risks to the relationship of Māori and their culture and traditions with their ancestral
lands, water, sites, waahi tapu, valued flora and fauna, and other taonga
iv.
the maintenance and enhancement of the capacity of people and communities to
provide for their own economic, social, and cultural wellbeing and for the reasonably
foreseeable needs of future generations
3.
Animal welfare and ethical issues.
4.
The benefits to be derived from the application.
Adequacy of the Proposed Containment Regime
The Committee’s consideration of the adequacy of the proposed containment regime, included:
1.
the ability of the organism or any heritable material to escape from containment
2.
the ability of the organism to establish a self-sustaining population
3.
the ease of eradication of any population established.
The ability of the organism or any heritable material to escape from
containment
In considering the ability of the organism to escape from containment, the Committee
considered inter alia, the following matters:
i.
escape of whole sheep
Page 8 of 36
ii.
escape of genetic material by horizontal gene transfer
iii.
disposal of sheep and other biological material
Sheep under this approval will be maintained in two separate containment facilities, initially in an
indoor facility (comprising two non-contiguous rooms), and later in an outdoor containment facility,
‘in the field’.
The Committee was satisfied, subject to the controls imposed in this decision, that the
containment regime can adequately contain sheep as a part of this field test. The controls require
sheep to be produced and maintained in registered containment facilities, constructed, operated
and managed in accordance with the MAF Biosecurity Authority/ERMA New Zealand
Standard154.03.06: Containment Standard for Field Testing of Farm Animals (outdoor containment
facility), and the MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.03:
Containment Facilities for Vertebrate Laboratory Animals (indoor containment facility), and to comply
with the controls specified in this approval.
Furthermore, an additional measure has been put in place over and above the requirements of the
standards, to further reduce the probability of any escape of sheep from the outdoor containment
facility by the installation of a system to electronically monitor the perimeter fencing in order to
promptly detect any interference or break in the fence.
The Committee was satisfied that the management and operational procedures in place are
sufficient to ensure that the sheep will be maintained in secure containment.
Under this regime, the Committee concludes that the likelihood of an escape from the facilities is
very low.
i.
Escape of whole sheep
Human Intervention
The outdoor and indoor containment facilities are located within the AgResearch RRC, and will
therefore be under constant close supervision.
The outdoor containment facility, originally registered as a 4ha facility, has been expanded and
now comprises a 45ha block. The extended facility has been approved by MAF under the relevant
MAF Biosecurity Authority/ERMA New Zealand Standard, and the Committee considers that it
provides equivalent containment to the previous 4 ha facility. In order to provide additional security
for the outdoor facility the applicant has installed further electronic monitoring systems, and access
to the facility is restricted to authorised persons through electronic card access. The electronic
systems in place are monitored outside of working hours by an external security provider, enabling
immediate detection of any breach of the perimeter fences.
For any potential saboteurs to gain access to the outdoor containment facility they would need to
breach a number of levels of security. They would need to gain access to the RRC (which is
monitored outside of working hours by a security provider on-site), locate the containment
facility, gain access to the facility without triggering the security system, and exit without being
detected.
The indoor containment facilities are located within an existing animal handling facility building,
located within the RRC complex, and the sheep will be housed in individual pens. Access to
these facilities is restricted to authorized personnel involved with the trial. Outside of working
Page 9 of 36
hours the facility is locked and infra-red detector beams are in place to detect any breach in the
outer security fence monitored by an external security provider.
For any potential saboteurs to gain access to the indoor containment facility, they would also
need to breach a number of levels of security. They would need to gain access to the RRC, locate
the building in which the indoor facility is located, gain access to that building without triggering
the security system, locate the containment facilities within the building, gain access to the
facilities themselves (past infra-red detector beams), and exit without being detected.
Identification measures in place for genetically modified sheep make sheep in this trial readily
identifiable, and distinct from other sheep within the RRC. The identification measures include
double tagging of all sheep, comprising either two ear tags (different in appearance to those in
common use on the surrounding farm) or an ear tag and a sub-cutaneous microchip, or for rams,
an ear tag and a permanent fixed radio tracking device capable of allowing remote detection and
location of the animal.
The Committee is satisfied that the construction, operation and management of the containment
facilities minimises the likelihood of any deliberate action or sabotage resulting in a breach of
containment, taking into account the security monitoring in place, fencing and access restrictions
and location of the facilities within the RRC.
In addition the Committee is satisfied that the operational and management procedures in place
minimise the likelihood of inadvertent release of organisms by personnel managing the trial.
Force Majeure
The Committee is satisfied that the containment regime proposed by the applicant (including
compliance with the MAF Biosecurity Authority/ERMA New Zealand standards and the
controls in this decision) is adequate to ensure that genetically modified sheep can be contained
in the event of an incident of force majeure, such as natural disaster, including earthquake, flood or
fire.
The site of the outdoor containment facility has been inspected by MAF and deemed to be a
suitable site for the purpose of maintaining genetically modified farm animals, with the aim of
ensuring containment.
In a force majeure event the applicant is required to take immediate steps to secure the facility, and
retrieve any animals that may have escaped from the facility. The containment standards for
registration of facilities require containment manuals that detail procedures that shall be followed
in an event of force majeure.
ii.
Escape of genetic material via horizontal gene transfer
Submitters have raised a number of concerns, in respect of this and previous applications, with
respect to the occurrence and purported consequences of horizontal gene transfer. This includes
such issues as:

the spread of foreign genes to unrelated species by ‘infection’ (horizontal gene transfer)

the potential for transferred material to create new viruses and/or diseases

the potential for transferred material to contribute to the spread of antibiotic resistance.
Page 10 of 36
Potential for the foreign DNA to move within the genome
Concern was expressed by some submitters that the E. coli gene used had an inherent capacity to
move within the genome. The Committee notes that this is not the case. A series of experimental
steps are needed to introduce this gene into the sheep because it is unable to insert by itself. The
antibiotic resistance gene was selected because it can be used as a selectable marker, not because
it can insert or move within the genome. The DNA derived from the E. coli bacterium and
introduced into the sheep myostatin gene has no inherent capacity to move between species, or
to move to other locations within the genome, because it does not have the genetic elements
necessary for transferring itself. Genetic elements that confer the ability for independent
movement are well characterised and distinctive, and the DNA to be introduced into the sheep
myostatin gene does not contain such sequences, nor will such mobility elements be created by
the insertion of the antibiotic resistance gene.
Horizontal Gene Transfer
Under certain conditions genetic material may be able to be transferred to other organisms by
non-reproductive means. Such examples of exchange of genetic material are called horizontal
gene transfer. These can most commonly occur via bacterial matings and viral infections.
Antibiotic resistance is transferred when resistance genes are on plasmids and these plasmids are
passed to other cells when bacteria mate (‘conjugate’). In the laboratory transfer of DNA to
bacteria requires specialised vectors and/or physiological conditions. If host DNA is transferred
by viruses this material is usually random pieces of DNA around the site of virus insertion and is
not specific to functional genes. The Committee notes that if horizontal gene transfer between
microorganisms, plants and animals was widespread and frequent then most individuals would
contain within their genomes a large diversity of genes and other pieces of DNA from a large
range of other organisms. This is not the case and reflects the fact that horizontal gene transfer is
generally restricted.
For the antibiotic resistance gene (or the disrupted sheep myostatin gene) to move to another
organism a series of independent steps would be required. For example, for the gene to be
acquired and expressed by a bacterium in the gut of the animal the following minimum number
of independent steps would be required:
i.
resistance to degradation of the gene in the sheep digestive system
ii.
uptake of a functional copy of the gene by a bacterium
iii.
integration of the gene into the bacterial genome (either into the chromosome or into a
plasmid)
iv.
expression of the gene.
Similar multiple steps would be required for other potential transfer pathways (such as transfer
to biting arthropods, or transfer via faecal material to soil microbes). Based upon published
scientific investigations it is very unlikely that any of these steps would occur. Calculating the
likelihood of the transfer path being completed requires the multiplication of each of the
individual step probabilities. The likelihood of such a transfer is, therefore, very much lower than
any of the individual steps, and so the Committee considers that the risk of horizontal gene
transfer by these means is negligible. Gut bacteria are continually exposed to cellular and DNA
material from their hosts with no evidence that the bacteria take up and incorporate host DNA.
DNA transfer between some gut bacteria has been demonstrated, but this involves direct
transfer of plasmids between the bacteria.
Page 11 of 36
iii.
Disposal of sheep and other biological material
The focus of the controls to which this approval is subject, is the containment of all genetically
modified material, either within the containment facility itself or on-site2 within the RRC.
Furthermore the controls require that all other biological material be disposed of on-site.
Although the risk of the transfer of genetic material by other than reproductive mechanisms is
considered to be negligible, this approval requires that all biological material be disposed of onsite.
This approach provides assurance that all sheep used in the field test, and all biological material
will be disposed of in an appropriate manner, and that this is able to be verified through the
records and registers the applicant is required to maintain.
For a small-scale field test, there are no practical impediments to on-site disposal, and for these
reasons controls relating to various components of biological material all require disposal on-site.
In addition, controls require the applicant to engage in on-going consultation with Ngāti Wairere
to develop culturally appropriate mechanisms and protocols for disposal of genetically modified
sheep and other biological material.
Disposal of sheep
Controls on this decision require that all sheep involved in the field test be disposed of, when no
longer required, on-site in accordance with the provisions of the MAF Biosecurity
Authority/ERMA New Zealand Standards 154.03.06 or 1.54.03.03 (as specified in control
number 1.12). This includes genetically modified sheep and foetuses, surrogate ewes that have
borne genetically modified lambs, conventional surrogate ewes that have failed to become
pregnant following embryo transfer, and non-genetically modified progeny.
For the purposes of publication and verification of any research results, the applicant may retain
semen and/or ova from genetically modified sheep following the conclusion of the field test.
Any semen or ova retained shall be held in accordance with the provisions of approvals to
develop the genetically modified organism (specifically embryos) and the control conditions to
which these approvals are subject.
Disposal of milk
The genetic modification introduced into sheep in this field test involves the disruption (knock
out) of the ovine myostatin gene locus. The modification involves the introduction of an
antibiotic resistance marker gene, and its protein product will be expressed. The genetically
modified (‘knocked out’) myostatin gene is under the control of the gene’s normal promoter and
so should demonstrate the same pattern and timing of expression as the unmodified myostatin
gene. The ovine myostatin protein is believed to be a negative regulator of muscle growth and
development, and is expected to be primarily expressed in the muscle tissue and blood of the
modified sheep. The antibiotic resistance gene is under the control of a promoter that is active in
most or all cells, and so may be expressed in other cells and tissues. Milk derived from genetically
modified sheep may, therefore, differ in composition from milk of conventional sheep, and
somatic cells carrying the modified (‘knocked out’) gene and marker sequences may also be
present in the milk of genetically modified sheep.
2
On-site refers to any place on AgResearch property within the Ruakura Research Centre
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Control conditions on this approval specify that all biological material, including waste milk and
cream, be disposed of on-site in accordance with the requirements of the MAF Biosecurity
Authority/ERMA New Zealand Standard 154.03.06 or 1.54.03.03.
Disposal of wool
The process of shearing sheep may result in some hair follicles or skin tissue being plucked, and
therefore shorn wool may contain DNA (including the modified DNA) adhered to a plucked
hair follicle.
Therefore, as for other biological products identified above, control conditions on this approval
specify that shorn wool, be disposed of on-site in accordance with the requirements of the MAF
Biosecurity Authority/ERMA New Zealand Standard 154.03.06 or 1.54.03.03.
The ability of the organism to establish a self-sustaining population and the
ease of eradication
Entry into the National Flock
Taking into account the containment regime it is considered very unlikely that a genetically
modified sheep might escape, enter the national flock undetected, and subsequently breeding
with conventional sheep in the national flock.
All sheep in this field trial, including conventional surrogate sheep, are required to be double
tagged at all times, comprising either, two ear tags (different in appearance to those in common
use on the surrounding farm), or an ear tag and a sub-cutaneous microchip. All genetically
modified ewes are required to carry both a visible ear tag and, at an appropriate age, a subcutaneous microchip. Prior to microchip implantation genetically modified lambs will be double
tagged, with two different ear tags, or have a visible tattoo.
Furthermore, in recognition of the fact that in the event of an escape, a ram poses the greatest
risk, in terms of potential to interbreed with conventional sheep in surrounding land, all
genetically modified rams are required to wear a permanently fixed radio tracking device capable
of allowing remote detection and location of the animal.
All sheep will therefore be readily identifiable in the event of an escape from containment.
Controls, to which this approval is subject, require that in event of unintended or accidental
release or escape of genetically modified sheep, that the applicant shall recover the escaped
sheep. Furthermore, if there has been any possibility of mating occurring, any possible resulting
pregnancies will be aborted, and if not successful, the affected sheep shall be slaughtered and
disposed of on-site. Alternatively, any potentially affected ewes shall be identified, destroyed, and
disposed of in accordance with the provisions specified in the controls.
Establishment of a Feral Flock
Although sheep are domesticated animals, feral sheep populations do exist in New Zealand. As
discussed above escape is considered to be unlikely, because of the nature of the containment
regime, and an undetected escape is considered to be even more unlikely because of the
proposed management regime. RRC is located adjacent to Hamilton City in a well populated
area, and therefore should an undetected escape occur it is very unlikely that the sheep reach a
suitable habitat to establish a feral flock and remain.
Page 13 of 36
The Committee therefore concluded that in the event of an escape from containment, it is highly
likely that sheep would be identified and retrieved. Furthermore, if there were a prospect that
escaped sheep had mated, any affected ewes would be able to be destroyed, the cost of which
would be borne by the applicant.
Effects of the Organism
Risk to Public Health
Concerns with respect to public health have been raised on a number of grounds, including:
i.
direct effects as a result of consumption of meat, milk, or offal derived from genetically
modified sheep
ii.
development of antibiotic resistance as a result of horizontal gene transfer to gut bacteria
iii.
unknown outcomes, e.g. CJD, and long-term unanticipated health effects.
i.
direct effects as a result of consumption of meat, milk or offal derived
from genetically modified sheep
The key issue is that of whether human consumption is likely to occur at all. The flock to be
produced under this approval is to be maintained in containment, and there is no intention for
the meat, offal or milk from genetically modified sheep, or sheep that have given birth to
genetically modified lambs, or surrogate ewes that fail to become pregnant, to enter the human
food chain.
The myostatin knockout sheep are expected to lack the ability to produce biologically active
myostatin protein, although they will be expected to produce the antibiotic resistance protein
product. In all cases the meat and milk will contain some modified genetic material, as the
transgenes will be present in every cell of the sheep.
As regards the potential harm to human health resulting from human ingestion of meat, offal or
milk from myostatin knockout sheep should it occur, the Committee identified two issues.
Firstly, the absence of a normal sheep myostatin protein was considered extremely unlikely to
present any ill effects if products from myostatin-deficient sheep were consumed. This is based
upon the fact that no altered sheep protein is produced. Furthermore myostatin-defective cattle
are consumed without any apparent ill effects. The Committee did, however, take account of the
fact that the presence of an antibiotic resistance protein product may provide some potential risk
of allergenicity if consumed. There are no published reports indicating that the neomycin or
puromycin resistance gene products are allergenic to humans.
While the applicant has no intention of producing food products as a result of this field test, if in
the future food products were to be developed from myostatin knockout sheep, further approvals
from ERMA, and approval from the Australia New Zealand Food Authority (ANZFA), would
be required before production of products for release could be undertaken.
Allergenicity
The neomycin resistance and puromycin resistance gene products do not pose any significant
allergenic concerns. In particular the neomycin resistance gene and its product has undergone
detailed biosafety evaluation. Human exposure to the protein without discernible effects on the
Page 14 of 36
immune system is assumed to be already occurring given the background of neomycin resistance
among microorganisms in the environment and human gut.
Most human allergens have a common set of characteristics (eg similarities in amino acid
composition, resistance to degradation in the gut) that enables many potentially allergenic
compounds to be identified. As the antibiotic resistance genes (both neomycin and puromycin)
to be used in this trial already occur in gut microorganisms without any apparent allergenic
effects it is not anticipated that there will be an allergenic response to them.
ii.
development of antibiotic resistance as a result of horizontal gene
transfer to gut bacteria
Submissions raised the prospect of the development of widespread antibiotic resistance as a
result of horizontal gene transfer following ingestion of products derived from genetically
modified sheep. The concern arises out of the fact that the genetic modification will introduce an
antibiotic resistance gene into the sheep genome. Unless removed the gene (and its product) is
expected to be present in the meat, milk and offal of the sheep developed using the selection
marker. However, expression of this product in some cells or tissues may be variable, very low,
or absent due to the fact that gene expression is also influenced by the local chromatin structure
(proteins which bind up the DNA in the chromosomes).
As noted above, the likelihood of consumption of products derived from genetically modified
sheep is considered to be very low, as is the likelihood of horizontal gene transfer given the
complexity of steps required to achieve it.
Even if such transfer of an antibiotic resistance gene to microorganisms, or to epithelial cells, did
occur in the gut the Committee considered that, set against an existing high background level of
antibiotic resistance (amongst microorganisms within the gut), the incremental consequence of
any transfer would be minimal.
However, in the unlikely event that such a transfer did occur, there is the potential for the
antibiotic to be inactivated if taken orally at the same time as meat derived from genetically
modified sheep or milk was consumed. The enzyme produced by the resistance gene that is
responsible for inactivating the antibiotic is however unlikely to be still present in food if it has
been cooked/heated. In addition in the case of neomycin it is poorly absorbed by mouth and is
given orally only in certain circumstances to reduce gut microorganisms eg preoperative bowel
preparation, hepatic encephalopathy. Such patients have dietary restrictions and are very unlikely
to be eating meat.
Therefore, taking into account the low likelihood of consumption of such products, and the low
likelihood of horizontal gene transfer actually occurring, the risk to human health as a result of
the development of antibiotic resistance is considered to be negligible.
iii.
unknown outcomes, ie CJD and long-term unanticipated health effects
Escherichia coli
The Committee notes that there was concern expressed by submitters regarding the use of a
gene derived from the bacterium Escherichia coli, and the potential for it to lead to disease or
genetic instability in the sheep. The gene involved is the neomycin antibiotic resistance gene (or,
alternatively, the puromycin antibiotic resistance gene), which is used both as a selectable marker
for the genetic modification and also as the means for disrupting the sheep myostatin gene.
Page 15 of 36
Some strains of E. coli, notably strain O157:H7, are known to cause acute diarrhea. This
pathogenicity is caused by specific toxins produced by these bacterial strains. In the case of
O157:H7 these are cytotoxins (called Shiga-like toxins or verotoxins) that are the products of
specific genes carried by this strain. The E. coli strain used in the myostatin research is not a
pathogenic strain and does not contain the verotoxin or other toxin genes. The antibiotic
resistance gene derived from the bacterium and introduced into sheep does not produce a toxin
that causes diarrhea or other diseases in humans; its function is to break down a specific
antibiotic compound. Products of this breakdown are not known to be toxic or to have other
detrimental effects on human health. The Committee therefore, considers that there is a
negligible risk for this E. coli gene to cause disease when introduced into sheep.
Creutzfeldt-Jakob Disease (CJD)
Submissions also raised the prospect that water can be a source of infection for CJD, and that
DNA could enter water supplies and provide such an opportunity. This issue is not relevant in
the context of this application, as none of the sheep will have scrapie or other prion diseases.
Prions (which cause scrapie, CJD, etc) are infectious misfolded proteins derived from specific
genes in the genome. Prion proteins all appear to share a similar set of amino acids, so that the
potential for a protein to exhibit prion-like activity can be readily identified. The genetic
modifications introduced in this application do not contain these prion-like elements. Myostatin
and the antibiotic resistance genes have not been shown to have any association with
susceptibility to these diseases. Therefore, the Committee considered that there is negligible risk
of the development of prion-like diseases from these modifications.
Risks to New Zealand’s ‘clean green image’, export relationships
and organic farming
Submitters raised the prospect that the presence of genetically modified sheep (including their
maintenance in containment) could jeopardise New Zealand’s expanding organic agricultural and
horticultural industries, and its ‘clean green image’.
In particular the concern expressed was that this trial, and others like it, may jeopardise New
Zealand’s ability to produce certified organic produce, and tarnish our ‘clean, green image’, and
that this poses a risk to the capacity of people and communities to provide for their own economic, social and
cultural well-being and for the reasonably foreseeable needs of future generations (section 5(b) Hazardous
Substances and New Organisms Act 1996).
Although the Committee does not dismiss such concerns, it considers that they are more
relevant to a release application (or the release of products) than to an application covering the
conduct of research in containment.
The Committee considers that so long as it is satisfied of the adequacy of the containment
controls and management regime, the risks are negligible for current and future generations alike.
Animal Welfare and Ethical Issues
A number of issues related to animal welfare were raised at the hearing. The focus of these
concerns was primarily that research aimed at the development of sheep in which muscle growth
is not controlled by myostatin may result in “uncontrolled” muscle growth with consequent
suffering and deformities for lambs and suffering for their mothers as a result of:
Page 16 of 36
(a)
birthing difficulties, resulting in turn in a higher than usual proportion of deliveries by
caesarean section
(b)
physiological effects such as difficulties in walking as a consequence of heavy muscling
(c)
unanticipated morphological effects, including intolerable deformities.
The applicant is required to comply with the relevant sections and regulations of the Animal
Welfare Act 1999, and the Animal Welfare Advisory Committee (AWAC) and National Animal
Ethics Advisory Committee (NAEAC) guidelines administered by MAF.
The Animal Welfare Act anticipates that animals will be used in research and may experience
some suffering in the process. The Act recognises that manipulation of a small number of
animals may result in benefits to a wider group of people or animals, to society generally, or to
the environment. The Act manages the use of animals in research and provides for the use of the
minimum number of animals required to maximise the benefits, whilst minimising the suffering
to the animals.
Any organisation using animals in research must maintain a Code of Ethical Conduct (CEC)
approved by MAF setting out policies and procedures to be followed by the organisation and its
Animal Ethics Committee. Furthermore every project involving the manipulation of animals,
including those undertaken in the course of the development and field-testing of genetically
modified sheep covered by this application must be approved by an Animal Ethics Committee
(AEC). Normally, where approval has been given the Authority should be able to assume that
all animal welfare matters of concern to submitters have been considered and addressed in that
approval.
In the present instance this was not evident from the original application, and did not become
clear until the Committee made further inquiries. From the additional information received,
however, it is evident that the Ruakura Animal Ethics Committee was fully seized of the
concerns expressed by submitters when it considered the programme, and that the ongoing
monitoring required to be exercised over the research in terms of the AEC’s approval is designed
to ensure that harm to lambs and their mothers is avoided or minimised.
In particular it is the expectation of the Ruakura AEC that the project will involve an expert
supervising large animal veterinarian to monitor the progress of lambs in utero, and take steps to
terminate a pregnancy and/or determine a humane end point where suffering to any animal is in
prospect. The Committee concluded that this requirement should be made a control condition
of its approval.
In addition, other controls to which this approval is subject, require the applicant to report to the
Authority on a regular basis from an animal welfare perspective on issues including the number
of and reasons for caesarean sections performed for genetically modified lambs; any
physiological effects relating to the heavy muscling of genetically modified sheep; and any
unanticipated behavioural or morphological effects. Finally, the applicant is required to forward
to ERMA New Zealand any reports provided to the relevant Ruakura AEC in respect of this
work.
A submitter raised the legal argument that, as the potential for suffering and deformity would
have been created at the time the embryos were modified by knocking out the myostatin gene,
this would have preceded the point at which the approval of an Animal Ethics Committee is
required. The Animal Welfare Act 1999 specifies that ethics approvals are only required in
respect of foetuses more than half way through their term. Animal ethical issues, it was argued,
Page 17 of 36
should have been considered at the time approval was sought to modify the embryo and, if the
applicant’s Animal Ethics Committee was not involved at this point because its jurisdiction was
not triggered, then consideration should have been given to the issues by the IBSC and explained
in its decision.
The majority of the Committee considered that as a practical matter, the Animal Ethics
Committee did address all the issues arising in consequence of the embryos having been
modified, even though this was in the context of an application for AEC approval to insert the
embryo into the sheep, and after the approval to modify the embryo had been given.
Nonetheless, there is a jurisdictional gap here, which until addressed by the Government, needs
to be handled by the Authority being able to assure submitters that their concerns have been
addressed by the relevant Animal Ethics Committee.
Benefits
The Committee noted that the principal benefit, as with all research, is the scientific information
expected to be gained - in this case from the development of genetically modified sheep based
on transfers of genetically modified embryos, with the result, if successful, that the sheep will not
express biologically active myostatin, and therefore may contribute to an increased understanding
of myostatin function and its effects on sheep muscularity.
For the longer term there may be other benefits, but these will only materialise as a consequence
of further research and development, and at this point it would be premature to speculate on
what those benefits might be.
Indeed, the majority of the Committee acknowledges that the degree of long term benefit to be
derived from this research, as with all fundamental research, is difficult to quantify. However,
that is not to say that the knowledge accumulated in the research is not beneficial as that
information adds to the pool of knowledge from which other benefits flow, and the Authority
accepts that exploratory research is an essential prerequisite for scientific progress.
Immediately, and for the purposes of most containment applications therefore, the issue is not
so much whether the longer term benefits outlined will be achieved, but whether research
leading to those potential benefits is a legitimate and valuable scientific endeavour.
This programme, to produce, and evaluate the muscle physiology and biochemistry of myostatin
knockout sheep, has been endorsed for funding by the Foundation for Research, Science and
Technology (FRST). The majority of the Committee accepts that given the significance of the
agricultural and pastoral industries in New Zealand, its research institutions should be at the
leading edge of research into the genetic factors which control and regulate muscle structure and
function, and of associated biotechnological innovation, including recombinant research, and
technologies of nuclear transfer (cloning) and surrogacy.
Risks to the Relationship of Māori and their Taonga
The applicant conducted consultations with Te Kōtuku Whenua, the environmental agency of
Ngāti Wairere, the Tainui hapu which claims mana whenua over the land on which the
applicant’s research facilities are located; Waikato Raupatu Lands Trust which also has
responsibilities for the land in question; Ngā Mana Tōpu O Kirkiriroa, which comprises all
senior kaumātua in the region, and is recognised by all the territorial local authorities in the
district on treaty matters; and with Hare Puke who is a kaumātua of Ngāti Wairere and kaumātua
to the applicant.
Page 18 of 36
It was evident that a diversity of views towards the application exists within the various bodies
consulted with some having no major issues with it.
No submissions on the application were made by Ngāti Wairere, but the applicant appended to
the application a Memorandum from Te Kōtuku Whenua containing a statement of Ngāti
Wairere’s views towards the application and a number of recommendations which may be
summarised as follows:
Ngāti Wairere considers that the alteration of the whakapapa of species is culturally
inappropriate and inherently against their tikanga, and that the Authority and the applicant have
a statutory duty under Te Tiriti O Waitangi to afford protection to whakapapa as a taonga. If the
proposed research involves alteration to normal growth patterns and therefore to the whakapapa
of the sheep, with potential risks to the wellbeing of the animals involved, Ngāti Wairere are
strongly opposed to the research proceeding. The consequences of proceeding include human
health risks, metaphysical imbalances and adverse psychological and physical effects, enduring
into the future, for both the mana whenua and all associated with the research or proximate to
the land on which this is conducted.
If the Committee should nonetheless decide to approve the application then Ngāti Wairere
recommends that every effort be taken to ensure that:

offal holes are designed in a culturally suitable manner as to prevent any potential risks on
the receiving environment;

an assessment of the potential risks and benefits to the cultural well-being of Ngāti Wairere
be undertaken by personnel agreed between Ngāti Wairere and the applicant.

a contingency plan for escape from the containment facility be established including
electrification of the inner fence;

a monitoring programme be established with representatives of Ngāti Wairere, with
provisions covering the avoidance, reduction or mitigation of any adverse effects detected in
consequence of the monitoring activity;

ongoing consultation with Ngāti Wairere representatives continue throughout the trial.
The Report of Ngā Kaihautū Tikanga Taiao, the Authority’s advisory body on matters relating to
sections 6(d) and 8 of the HSNO Act, after reiterating the points made by Te Kōtuku Whenua
consultants, commented as follows:
Ngā Kaihautū takes into account the need to be mindful of its previous decisions. It takes the
view that unless there is compelling evidence to the contrary, it should maintain a position that is
not clearly inconsistent with these. This view is reinforced by the knowledge of an impending
Royal Commission and ERMA New Zealand’s own Generic Issues research project. Until these
have provided both ERMA New Zealand and Ngā Kaihautū with a more informed
understanding of the issues of concern to Māori, we will maintain our support for the
precautionary approach as it pertains to both science and ethics concerning this type of research,
particularly in organisms intended for eventual field release and/or possible human
consumption.
In view of the above considerations, Ngā Kaihautū Tikanga Taiao recommends that the
Committee decline this application.
Page 19 of 36
Similar issues to those raised by Te Kōtuku Whenua and Ngā Kaihautū arose in the application
considered by the Authority (GMF98009) to field test cattle developed from embryos into which
had been inserted a copy of gene coding for the human protein myelin. That case was regarded
more seriously by Ngāti Wairere in that it involved the transplantation of a gene coding for
human protein, whereas the present application involves the knockout of a gene – although in
both cases the point at issue is the intereference with the whakapapa of the manipulated species.
And in the present instance, the recommendations made by Ngāti Wairere, through the
applicant, contemplate the possibility of the Committee approving the application.
The Committee acknowledges the seriousness of the matters raised by Te Kōtuku Whenua on
behalf of Ngāti Wairere and has weighed them carefully. As with the cattle decision however,
the majority of the Committee did not consider that these matters were such as to justify
declining the application. In so concluding, the majority noted in particular that there was a
diversity of views towards the application amongst those consulted by the applicant.
Most of the matters contained in the recommendations of Ngāti Wairere regarding the disposal
of offal, containment, monitoring and consultation have been addressed in the controls attaching
to this decision.
As regards the health risk assessment sought by Te Kōtuku Whenua, the majority of the
Committee does not believe it is practical to require that this be undertaken as a condition of the
approval, but it is of the view that if Ngāti Wairere choose to undertake the assessment
themselves, the applicant might look for ways of providing practical support.
Overall evaluation of risks, costs and benefits and
conclusions
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)(b): Field testing any new
organism.
The Committee is satisfied that the proposed containment regime, together with the additional
controls imposed by the Committee, will adequately contain the genetically modified sheep and
any heritable material. The likelihood of escape from the facilities is very unlikely, and the
likelihood of any escape going undetected with consequent failure to retrieve the escaped animals
and take action to manage any possible matings that have occurred, is considered to be even
more unlikely. In recognition that rams pose the greatest risk in the event of escape from
containment, the controls require all genetically modified rams to wear a permanently fixed radio
tracking device allowing remote detection and location of the animal.
The Committee concludes that risks to the environment and human health from the possible
escape of the genetically modified sheep are negligible, given the nature and extent of the
containment and sheep management regime set out in this approval.
The Committee gave particular consideration to animal welfare issues. The Committee notes the
jurisdictional gap that exists in the Animal Welfare Act 1999 in relation to foetuses that are less
than half way through their term. Nevertheless the Committee is satisfied that the Ruakura
Animal Ethics Committee has considered all the issues that arise in consequence of the embryos
being modified. Controls on this application ensure that the husbandry of the animals shall be
overseen by an experienced large animal veterinarian who has authority to terminate the
pregnancies or the trial, should it be necessary to avoid unacceptable suffering.
Page 20 of 36
The majority of the Committee concludes that the proposed research for which this field trial
application is sought, is a legitimate and valuable scientific endeavour. Research results may
contribute to an increased understanding of myostatin function and its effects in sheep
muscularity, and contribute to a wide range of related research. The majority of the Committee
accepts that the information can only be obtained efficiently from knockout procedures, noting
that heavy muscled sheep such as Texels and Bextels are not known to have defective myostatin,
and would therefore not be appropriate for this research. Further, given the significance of
agricultural and pastoral industries in New Zealand, its research institutions should be at the
leading edge of research into genetic factors that control and regulate muscle structure and
function, and of associated biotechnology techniques. The majority of the Committee also
accepts that there are scientific benefits to be gained from research using larger animals such as
sheep (in which the applicant has considerable experience), in addition to smaller laboratory
animals.
The Committee acknowledges the seriousness of the matters raised by Te Kōtuku Whenua on
behalf of Ngāti Wairere and has weighed them carefully. The Majority of the Committee did not
consider that these matters were such as to justify declining the application. In so concluding,
the majority noted in particular that although it have given particular attention to the views
expressed by Ngāti Wairere through the Te Kōtuku Whenua there was a diversity of views
towards the application amongst those consulted by the applicant. Most of the matters
contained in the recommendations of Ngāti Wairere regarding the disposal of offal, containment,
monitoring and consultation have been addressed in the controls attaching to this decision.
Minority Decision
The minority considers that the application should be declined on three grounds:
First, because of the adverse effects it may have on Ngāti Wairere, approval of this application
imposes yet another affront to these people already burdened by concerns about a previous
application. The cumulative effect is likely to contribute to increased stress on their spiritual and
physical well being.
Secondly, on the grounds that the consultation process did not provide a sufficiently robust basis
on which to properly evaluate and weigh the risks, effects and benefits to tangata whenua. The
minority argues that a process in which the applicants themselves conduct the assessment of
cultural effects is inappropriate and seriously flawed. Instead it is suggested that assessment of
cultural effects should in future be conducted or facilitated by an independent and appropriately
qualified person or persons - a situation which is likely to engender more trust and willingness by
iwi to engage in a fully informed way in the process. An additional problem was the conflation
of this application with a previous one (GMF98009) a situation which, by their own admission,
led tangata whenua to give inadequate time and attention to this particular proposal. Of concern
also was the potential for conflict of interest among the source of advice sought by the applicant
on cultural risks and effects.
Thirdly, because the proposed benefits of the scientific information to be obtained from this
research do not outweigh the possible adverse effects on the health and well being of the animals
involved in the programme. The minority believes, as a general principle, that approval should
only be given if the outcomes are clearly perceived to be of benefit to society, and/or where
there are considered to be no viable alternatives to the research.
The application for the field testing of myostatin knockout sheep is thus approved with controls.
Page 21 of 36
Controls
In order to provide for the matters detailed in Part I of the Third Schedule to the Act, Containment
Controls for Development and Field Testing of Genetically Modified Organisms, 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 material3:
1.1
The applicant before field testing sheep containing any construct not yet developed, shall
obtain development approval, under the Hazardous Substances and New Organisms Act
1996, from the AgResearch Institute’s Ruakura Institutional Biological Safety Committee
(IBSC) and provide a declaration in writing to the Authority verifying that:
1.1.1
the construct and genetically modified embryo has been developed in accordance
with an approval under section 39(1)(a) of the Act
1.1.2
the construct and genetically modified embryo complies with the requirements
detailed in the attached schedule to this decision
1.1.3
the genetically modified cell line (nuclear donor) from which the embryo is
produced contains the transgene (verified by methods including, but not limited
to, the Polymerase Chain Reaction (PCR) or Southern hybridisation analysis).
1.2
The operation, management and construction of the indoor containment facilities4 shall be in
accordance with the Ministry of Agriculture and Forestry (MAF) Biosecurity
Authority/ERMA New Zealand Standard 154.03.03: Containment Facilities for Vertebrate
Laboratory Animals.
1.3
The indoor facility shall be approved by MAF as a containment facility in accordance with the
MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.03 prior to housing of
any sheep under this approval.
1.4
The operation, management and construction of the outdoor containment facility shall be in
accordance with the MAF Biosecurity Authority/ERMA New Zealand Animal Health and
Welfare Standard 154.03.06: Containment Standard for Field Testing of Farm Animals.
1.5
The outdoor facility shall be approved by MAF as a containment facility in accordance with
the MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.06 prior to the
introduction of any sheep into the facility.
1.6
The production and maintenance of genetically modified sheep in the containment
facilities shall be in accordance with the relevant sections and regulations of the Animal
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
Containment facility means a place approved in accordance with section 39 for holding organisms that should not,
whether for the time being or ever, become established in New Zealand. Biosecurity Act 1993
4
Page 22 of 36
Welfare Act 1999, the Animal Welfare Advisory Committee (AWAC) and National Animal
Ethics Advisory Committee (NAEAC) guidelines administered by MAF, and the local
AgResearch Institute’s Animal Ethics Committee (AEC). The husbandry of these animals
shall be overseen by an experienced large animal veterinarian, who shall have the power to
determine a humane endpoint for any part of the trial or overall endpoint of the trial.
1.7
The maximum number of sheep in the field test shall not exceed the capacity of the
containment facilities, and/or any requirements of the relevant Animal Ethics Committee,
and should at all times be the minimum number of animals required to obtain statistically
significant results.
1.8
The maximum number of genetically modified sheep in the field test shall not at any one
time exceed 100 animals.
1.9
The identification system for genetically modified sheep shall enable information on the
genotype and generation (T0, T1 etc) to be derived from a database maintained by the
applicant.
1.10 The outdoor containment facility shall be enclosed by a double perimeter fence
constructed in accordance with the requirements specified in the MAF Biosecurity
Authority/ERMA New Zealand Standard 154.03.06. The inner perimeter fence shall be
electronically monitored and alarmed (in order that the location of any breach of
containment is detected immediately), stock-proof and capable of preventing entry and
escape of sheep.
1.11 No genetically modified sheep shall leave the containment facilities except in accordance
with the MAF Biosecurity Authority/ERMA New Zealand Standards, as described in
controls 1.2 and 1.4.
1.12 In the event that operations involving genetically modified sheep cease, all genetically
modified sheep in the containment facilities shall be destroyed and disposed of on-site in
accordance with the requirements of the MAF Biosecurity Authority/ERMA New
Zealand Standard 154.03.06, and of any requirements under the Resource Management
Act 1967, and in consultation with Ngāti Wairere with respect to developing culturally
appropriate mechanisms and protocols for disposal.
1.13 All sheep no longer required for breeding and any biological material (including semen,
ova, milk and shorn wool) derived from genetically modified sheep no longer required for
the purpose of this application shall be disposed of on-site5 in accordance with the
requirements of the MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.06,
and of any requirements under the Resource Management Act 1967, and in consultation
with Ngāti Wairere with respect to developing culturally appropriate mechanisms and
protocols for disposal. Ova and semen can be kept in secure containment for record
purposes.
1.14 The applicant shall provide ERMA New Zealand and the facility Supervisor6 (MAF) with a
timetable for the production and field testing of genetically modified sheep approved
5
On-site refers to any place on AgResearch property within the Ruakura Research Centre.
6
An Inspector appointed under the Biosecurity Act.
Page 23 of 36
under this decision, and shall notify ERMA New Zealand and the facility Supervisor, in
writing, of any changes to that timetable.
1.15 At all times only persons authorised by the Operator or the Manager of the containment
facilities shall have access to the containment facilities, other than those identified in
control 6.1
1.16 All sheep in the field test (whilst in the indoor and outdoor containment facilities) shall be
double tagged at all times.
1.16.1 All genetically modified ewes shall be individually identified by an ear tag for
visible identification and also implanted with a subcutaneous electronic microchip
for individual electronic identification. In the event that subcutaneous microchips
cannot be inserted until sheep reach a certain age, sheep should have two different
types of ear tag in place at all times, allowing for immediate identification.
1.16.2 All genetically modified rams shall be fitted with a permanently fixed radio
tracking device capable of allowing remote detection of the location of rams at all
times.
2. To exclude unauthorised people from the facility:
2.1
The applicant shall comply with the requirements contained in the standards listed in
control 1.2 and 1.4 relating to identification of entrances, numbers of and access to
entrances, and security requirements for the entrances and the facility.
3. To exclude other organisms from the facility and to control undesirable
and unwanted organisms within the facility:
3.1
The applicant shall comply with the requirements contained in the standards listed in
control 1.2 and 1.4 relating to exclusion of other organisms from the facilities and the
control of undesirable and unwanted organisms within the facilities.
3.2
In the event of mortality in genetically modified sheep in the field test site, carcasses shall
be immediately removed to prevent access to scavengers and following investigation as
required by Section 4.8 of The MAF Biosecurity Authority/ERMA New Zealand Standard
154.03.06, the carcasses shall be disposed of on-site in accordance with the provisions
specified in control 1.12.
4. To prevent unintended release of the organism by experimenters working
with the organism:
4.1
The applicants shall comply with the requirements contained in the standards listed in
control 1.2 and 1.4 relating to the prevention of unintended release of the organism by
experimenters working with the organism.
4.2
No part or product of the transgenic organism shall be ingested by any person at any time.
Page 24 of 36
5. To control the effects of any accidental release or escape of an organism:
5.1
In case of unintended or accidental release or escape of genetically modified sheep
involved in the field test, the applicant shall recover the escaped sheep to the containment
facility. If there has been any possibility of mating occurring, steps shall be taken to abort
any possible resulting pregnancies. If the abortion is not successful, the affected sheep
shall be slaughtered and disposed of on-site. Alternatively, any potentially affected ewes
shall be identified and destroyed, and be disposed of in accordance with the provisions
specified in control 1.12.
5.2
If for any reason a breach of containment occurs the applicant shall notify the facility
Supervisor (MAF) and ERMA New Zealand immediately the event is noticed (and at least
within 24 hours of the breach being detected).
6. Inspection and monitoring requirements for containment facilities:
6.1
The Authority or its authorised agent or properly authorised enforcement officers, or
supervisor under the Biosecurity Act 1993, may inspect the containment facility at any
reasonable time.
6.2
The applicant shall provide a comprehensive report to ERMA New Zealand in each
December on the progress in the production and field testing of genetically modified
sheep, including an inventory (a register that records the identity and fate of all sheep in
the field trial), with particular reference to the topics listed in section 4.13 of the MAF
Biosecurity Authority/ERMA Standard 154.03.06. This report shall also include:
6.2.1 information on animal welfare issues including; unexpected birthing difficulties,
unexpected mortalities and any unusual physiological, morphological, and
behavioural effects resulting from the genetic modification;
6.2.2 information on the stability of the genetic constructs used in genetically modified
sheep; and
6.2.3 any reports provided to the local Ruakura Animal Ethics Committee7.
6.3
7
The applicant shall provide a final report to ERMA New Zealand at the conclusion of the
approval period, being five years from the date of this decision. This shall include:
6.4.1
information on the items listed in section 4.13 of the MAF Biosecurity
Authority/ERMA Standard 154.03.06
6.4.2
information on animal welfare issues including; unexpected birthing difficulties,
unexpected mortalities and any physiological, morphological and behavioural
effects resulting from the genetic modification
6.4.3
information on the stability of the genetic constructs used in genetically modified
sheep
6.4.4
any reports provided to the local Ruakura Animal Ethics Committee.
Animal Ethics Committee
Page 25 of 36
6.4
The applicant shall establish an on-going liaison committee with Ngāti Wairere, to enable
Ngāti Wairere to monitor the implementation and progress of the field test, and to provide
a forum for the exchange of information on the science of genetic modification.
7. Qualifications required of the persons responsible for implementing those
controls:
7.1
The applicant shall comply with the requirements of the standards listed in control 1.2
relating to the training of personnel working in the facility.
7.2
The applicant shall notify the Supervisor and ERMA New Zealand if there are any changes
in ownership of property housing the containment facility maintaining organisms under
this approval.
_____________________
Chair
__________________
Date
_____________________
Name
Page 26 of 36
References
Fuchs RL, Ream JE, Hammond BG, Naylor MW, et al. Safety assessment of the neomycin
phosphotransferase II (NPTII) protein. Bio/technology 1993; 11: 1543-47.
Huby, R.D., Dearman, R.J., Kimber, I. (2000). Why are some proteins allergens? Toxicol. Sci. 55,
235-246.
Karenlampi S. Health effects of marker genes in genetically engineered food plants. Copenhagen:
Nordic Council of Ministers, 1996.
Mackenzie D. Gut reaction. Could a mechanical gourmet help us digest a GM future? New
Scientist. 30 January 1999: p4
Metcalfe DD, Astwood JD, Townsend R, Sampson HA, et al. Assessment of the allergenic
potential of foods derived from genetically engineered crop plants. Crit Rev Food Sci Nutr 1996;
36(S): S165-86.
US Food and Drug Administration. Guidance for industry: use of antibiotic resistance marker
genes in transgenic plants. Draft. Rockville: US FDA, September 1998.
US Registry of Toxic Effects of Chemical Substances
1997. Foreign (M13) DNA ingested by mice reaches peripheral leukocytes, spleen, and liver via
the intestinal wall mucosa and can be covalently linked to mouse DNA. Proc. Natl. Acad. Sci.
USA 94, 961-966
1999, Immunization via hair follicles by topical application of naked DNA to normal skin.
Nature Biotechnology 17 (870-872)
World Health Organization. Biotechnology and food safety: report of a joint FAO/WHO expert
consultation. Geneva: World Health Organization, 1996
Page 27 of 36
Schedule 1
Organism:
Sheep (Ovis aries)
Construct:
Myostatin knockout (disruption of the sheep myostatin locus by
the introduction of a neomycin or puromycin (antibiotic)
resistance gene.
Genetic modifications:
The constructs shall contain only genes, promoters and marker
sequences specified in the application.
Page 28 of 36
Annex 1
Details of the genetic modification
Gene Construct
The sheep myostatin locus with three exons and two introns has been cloned into pBlueScript.
The neomycin or puromycin resistance gene has been cloned into the 1st exon of myostatin. This
will enable the selection of homologous recombination events by resistance to G418 (Geneticin).
The diagram below illustrates the unmodified and modified myostatin gene.
The figure shows the genomic structure (panel A) and the knockout construct (panel B). The
myostatin gene contains three exons and two introns. The exons are denoted in light shading,
and the introns in dark shading. The nucleotide numbers corresponding to the exons are shown.
In the knockout construct the neomycin (neo) gene is inserted between nucleotides 330 and 331
of the 1st exon, and the thymidine kinase (tk) gene is cloned at the 3’ end of the myostatin gene.
The knockout construct is generated using pBlueScript (SK+). While the neomycin resistance
gene is used for positive selection of homologous recombinations at the myostatin locus, the tk
gene will be used as a negative selection marker for non-specific integration.
A
Myostatin Genomic structure
1
373
374
740
741
1128
B
Myostatin Knock out construct
-126
+330
331
neo
374
740
741
1128
tk
= EXON
= INTRON
Source of Nucleic Acid
The sheep myostatin locus has been PCR-amplified using genomic DNA from “Thomas”
(Cloned sheep) as a template. The neomycin8 and puromycin9 resistance genes used in this
project are synthetic sequences identical to those located in the E. coli genome.
Southern PJ and Berg P. 1982 Transformation of mammalian cells to antibiotic resistance with a bacterial gene
under control of the SV40 early region promoter. Journal of Molecular and Applied Genetics, 1 327 – 341.
8
De la Luna S, Soria I, Pulido D, Ortin J and Jiminez A. 1988 Efficient transformation of mammalian cells with
constructs containing a puromycin-resistance marker. Gene 62(1) 121 – 126.
9
Page 29 of 36
List of submitters
Annex 2
Page 30 of 36
Name
Organisation
Position
Address
Request
to be
heard
1
Susan Redward
Federated
Farmers of New
Zealand (Inc)
Policy Manager PO Box 715
2
Berylla Berylla
24 Montcalm Street
NELSON
3
Duncan Sloss
12 Newbury St
Opawa
4
Robert Anderson
440a Otumoetai Road
TAURANGA
5
Tim Vallings
Brit Rd
RD 8
WHANGAREI
Yes
6
Zelka Grammer
Bint Road
RD8
Maungakaramea
WHANGAREI
Yes
7
Susie Lees
14 Hastings Street
NELSON
8
9
Justine Gresson
Paul de Spa
50 Sheldon St
50 Sheldon St
CHRISTCHURCH
CHRISTCHURCH Yes
Yes
10
Norman Fletcher
PO Box 17
Waikawa Bay
PICTON
Yes
11
Wendy McGuinness
50 Cashmere Avenue
Khandallah
WELLINGTON
Yes
12
Tscherner Wolfgang
326 Princess Drive
NELSON
Yes
13
Pip Direen
50 Atkins St
MOTEUKA
Yes
14
Gina Luke
4 Landy St
Dallington
15
Genevieve Claire de
Spa
125 Kainga Rd
CHRISTCHURCH 4
16
Claire Bleakley
Pigeon Bush
Western Lake Rd
FEATHERSTON
Yes
17
Associate Professor
Peter Wills
Department of Physics
The University of
Auckland
Private Bag 92
019
AUCKLAND
Yes
18
V A Clayton
c/- Ginny's Herbs
PO Box 35
Maungakaramea
No
No.
WELLINGTON
Yes
Yes
CHRISTCHURCH Yes
No
Yes
CHRISTCHURCH Yes
No
Page 31 of 36
Name
Organisation
Position
Address
Request
to be
heard
No.
NORTHLAND
19
Carolyn Rhodes
32 Wembley St
CHRISTCHURCH
8002
20
Mike Drake
PO Box 10
Tapawera
21
Budyong & Lesley
Hill
11 Huiawa St
WAIKANAE 6454
No
22
Alan J Baldwin
94 Marne St
PALMERSTON
NORTH
No
23
Mike Aitkenhead
RD 1
Mangakino
No
24
Gilbert Urquhart
48 Glenroy St
CHRISTCHURCH
Yes
25
Dr Rebecca Potts
33 Wyndham St
AUCKLAND
Yes
26
Malcolm Frost
107 Havelock Road
Havelock North
HAWKES BAY
Yes
27
Elizabeth Frost
107 Havelock Road
Havelock North
HAWKES BAY
Yes
28
Rich J Wernham
54 Eden St
Island Bay
WELLINGTON
Yes
29
Mary Anne Howard- R.A.G.E Inc
Clarke
PO Box 30 762
LOWER HUTT
30
Helen Scott
185A Keyes Rd
New Brighton
CHRISTCHURCH Yes
31
Manfred von
Tippelskirch
185A Keyes Rd
New Brighton
CHRISTCHURCH Yes
32
Pat Lee
54 Waipori St
Berhampore
WELLINGTON
33
Don and Rose Craig Green Horizons
Organics Ltd
PO Box 113
Kirwee
CHRISTCHURCH Yes
34
Marcus Graf
24 Baildon Rd
Grey Lynn
AUCKLAND
Yes
35
Shannon McGhee
Pupuke Rd
RD 2
Kaeo 0471
NORTHLAND
Yes
36
Jon Muller
PO Box 39 158
WELLINGTON
37
Sam Stocker
54 Waipori St
Newtown
PPL Therapeutics
(New Zealand) Ltd
National
Spokesperson
Member for
R.A.G.E Inc
Yes
NELSON
Yes
Yes
No
Yes
WELLINGTON
Page 32 of 36
No
Name
Organisation
Position
Address
Request
to be
heard
No.
38
Jim Gladwin
22a Walsall St
Avondale
39
Tammy Meaglter
50 Atkins St
MOTUEKA
40
Sara Tree
35 14th Avenue
TAURANGA
41
Ngaire Irwin
77 Owairaka Ave
Mt Albert
42
Kelly Newman
PO Box 54
NELSON
Yes
43
Tahu Erlbeck
PO Box 54
NELSON
Yes
44
John Janke
Hamama Rd
RD 1
45
H J Clark
20 Bishopdale Ave
NELSON
Yes
46
Joanna M Paul
3 Maxwell Ave
WANGANUI
Yes
47
Oraina Jones
47 Ellis St
Brightwater
48
Sue Kedgley
Parliament Buildings
WELLINGTON
Yes
49
Sigi Kirchmair
9 Leach Place
NELSON
Yes
50
Alison Fletcher
PO Box 17
Waikawa Bay
PICTON
Yes
51
Graeme North and
Deniece Gannaway
Matthew Road
RD 1
WARKWORTH
Yes
52
Alex Vickers
Flat 1
44 Roxburgh St
Mt Victoria
WELLINGTON
Yes
53
New Zealand Life
Sciences Network
c/- Mr MRG
Christensen
Anderson Lloyd
Level 13, Forsyth Barr Cnr Armagh &
Yes
House
Colombo St
CHRISTCHURCH
54
Biotenz
C/- Mr MRG
Christensen
Anderson Lloyd
Level 13, Forsyth Barr Cnr Armagh &
Yes
House
Colombo St
CHRISTCHURCH
55
Raymond Vogt
89 Maungaraki Rd
Petone
56
Julia Bromley
177 Vanguard St
NELSON
Yes
57
Tejas Arn
177 Vanguard St
NELSON
Yes
58
Estelle Courtney
1 Boyes Place
NELSON
Yes
59
Marcia Higgs
c/- 11 Dundas St
NELSON
Yes
MP
AUCKLAND
Yes
Yes
Yes
AUCKLAND
TAKAKA
NELSON
WELLINGTON
Page 33 of 36
Yes
Yes
Yes
Yes
Name
Organisation
Position
Address
Request
to be
heard
No.
60
Nick Young
57 Wolfe St
NELSON
Yes
61
Kei Munro
385 Brook St
NELSON
Yes
62
Nicholas Owen
276 Hardy St
NELSON
Yes
63
Joanne Emery
94 Collingwood St
NELSON
Yes
64
Diana Kirpensteijn
5 Ombersley Tce
CHRISTCHURCH
No
65
Zanna Bird
64 Brougham St
NELSON
Yes
66
Andrea Chandler
33 Queens Rd
NELSON
Yes
67
Grant Scott
128 Grove St
NELSON
No
68
Paul Brumwell
East Takaka Rd
Takaka
GOLDEN BAY
69
Charles
Satterthwaite
35 Van Asch St
Sumner
CHRISTCHURCH No
70
Peter and Kerry
Green
PO Box 133
TAKAKA
71
Tracy Buick
68A Taupo St
Green Bay
72
Jeff and Karen Hay
PO Box 62
TAKAKA
Yes
73
Silvia Gassebner
9 Leach Place
NELSON
Yes
74
Bruce Dyer
PO Box 984
NELSON
No
75
Michael Glover
Waihora
Days Bay
RD 4
Yes
CHRISTCHURCH
76
Trudy Burgess
PO Box 693
NELSON
Yes
77
Jim Chapple
260 Willoughby Rd
RD 1
KATIKATI
78
Penelope Buxton
192 Clyde Rd
Fendalton
CHRISTCHURCH No
79
Peter Davis
11 Aorangi Tce
WELLINGTON
No
80
Noel Wierzbicki
RD 9
PALMERSTON
NORTH
Yes
GE Free NZ
National
Coordinator
Yes
No
AUCKLAND
Page 34 of 36
Yes
No
Annex 3
Qualitative scales for describing effects
Adverse effects
Qualitative assessments use words to describe the probability or likelihood of the adverse effect
occurring and the magnitude, or a measure of the severity of that adverse effect. These are
combined to form a qualitative estimate of the level of risk. The following sets of word scales
are used:
Table 1: Likelihood of effect
Descriptor
Description
Very unlikely
Not impossible, but only occurring in exceptional circumstances
Unlikely
Could occur, but is not expected to occur under normal
conditions
Possible
50:50 chance of occurring, or where there is insufficient
information to judge likelihood otherwise
Likely
Will probably occur at some time
Very likely (almost certain)
Is expected to occur
Table 2: Magnitude of adverse effect
Descriptor
Examples of descriptions
Minimal
Insignificant (repairable or reversible) environmental impact, no
observable cultural effects, other effects slight (reversible) or very small
Minor
Reversible environmental impact, limited adverse cultural effects
(affecting small area or localised community), other effects small and
limited in scope
Moderate
Some slight effect on native species, adverse cultural effects to wider
area but not considered serious, other effects medium or mid range
Major
Irreversible environmental effects but no species loss, adverse cultural
effects widespread but remedial action available, other effects large
Massive
Extensive irreversible environmental effects, including species loss,
adverse health effects, severe adverse cultural effects over whole
Page 35 of 36
country with no possible remedial action, other effects very large and
widespread
Beneficial effects
The word scale used to describe the likelihood of beneficial effects is the same as stated in
Table 1 above. A qualitative assessment to describe the magnitude of beneficial effects has been
made. The following word scale is used:
Table 3: Magnitude of beneficial effect
Descriptor
Qualitative description
Minimal
Very small or insignificant benefit
Minor
Small benefits that are localised in distribution
Moderate
Medium benefits that are localised in distribution
Major
Large benefits that are regional level in distribution
Massive
Very large benefits that are national in distribution
Ngā Kaihautū Tikanga Taiao has been formally established under clause 42 of the First Schedule to the Hazardous
Substances and New Organisms Act 1996, as a Māori advisory committee, to advise the Authority on how to take
account of issues of concern to Māori (particularly in relation to sections 6(d) and 8 of the Act).
i
Page 36 of 36