ENVIRONMENTAL RISK MANAGEMENT
AUTHORITY DECISION
Amended under s67A on 23 August 2007
16 January 2006
Application code:
GMD05066
Application
category:
Develop in Containment any New Organism under the
Hazardous Substances and New Organisms (HSNO) Act 1996
Applicant:
University of Otago
Purpose:
To generate genetically modified fungi and Eschericia coli in
order to study how antifungal drugs interact with their targets
and how fungal cells become resistant to drugs. This will assist
towards the development of better drugs, and solutions to
overcoming drug resistance.
Date application
received:
10 October 2005
Consideration:
6 December 2005
Considered by:
A Committee of the Authority
1
Summary of Decision
1.1
Application GMD05066 to develop in containment:
Escherichia coli (non-pathogenic strains), Candida albicans, Candida
dubliniensis, Candida glabrata, Candida krusei, Candida tropicalis,
Saccharomyces cerevisiae (non-pathogenic strains), Aspergillus fumigatus
and Cryptococcus neoformans, genetically modified by plasmid vectors or
E. coli/yeast shuttle vectors containing selectable markers1, reporter genes,
standard protein tags, regulatory sequences2 and donor DNA from the fungi
listed above. Donor DNA shall encode azole or echinocandin drug targets, or
genes involved in azole or echinocandin drug resistance.

Only one drug target or resistance mechanism will be investigated in
each modification.
1
Including antibiotic resistance genes and genes involved in nutrition biosynthesis to select for
genetically modified fungi.
2
Fungal genes will be expressed under the control of their own promoters or alternative fungal
promoters that give high or low levels of expression.
has been approved with controls, having been considered in accordance with
the relevant provisions of the Hazardous Substances and New Organisms
(HSNO) Act 1996 (the Act) and the HSNO (Methodology) Order 1998 (the
Methodology).
2
Legislative Criteria for Application
2.1
The application was lodged pursuant to section 40(1)(b) of the Act and
determined by the Environmental Risk Management Authority (the
Authority) in accordance with section 45, having regard to section 43 and to
matters relevant to the purpose of the Act, as specified under Part II. Unless
otherwise stated, references to section numbers in this decision refer to
sections of the Act.
2.2
The application does not qualify for rapid assessment under section 42 as it
involves a range of genetic modifications, some of which are prescribed as
not low-risk in clause 1(k) of the Schedule to the HSNO (Low-Risk Genetic
Modification) Regulations 2003: developments involving modifications to
pathogenic micro-organisms that result in resistance to antibiotics used for
clinical or veterinary treatment of infections caused by that micro-organism.
2.3
Consideration of the application followed the relevant provisions of the
Methodology, with particular regard to clauses 12 (dealing with assessment
of risks) and 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
Application Receipt
3.1
Application GMD05066 was determined to be in compliance with section
40(2) of the Act and was formally received on 10 October 2005.
Notification
3.2
The application was not publicly notified because the Authority considered
that there was unlikely to be significant public interest in this application.
3.3
The Ministry of Agriculture and Forestry (MAF) (Biosecurity New Zealand)
and the Department of Conservation (DoC) were notified of the receipt of
this application and provided with an opportunity to comment.
Information Available for Consideration
3.4
The information available for the consideration of the application was:
The application prepared by the applicant, including scientific references.
The Evaluation and Review (E&R) Report prepared by ERMA New Zealand including
comments provided by those government agencies notified of the application.
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3.5
The Authority has authorised the completion of the E&R Report as
additional information under section 58(1)(b) of the HSNO Act 1996. The
consideration of this application was postponed under section 58(3) of the
HSNO Act 1996, to allow for the completion of the E&R Report.
3.6
The E&R Report was provided to the applicant five working days prior to
the consideration and a response was obtained before the consideration.
Decision Making Committee
3.7
The application was considered by a Committee of the Authority, appointed
in accordance with section 19(2)(b) of the Act and clause 43 of the First
Schedule to the Act. For the purposes of determining this application, the
Committee comprised the following members: Dr Kieran Elborough (Chair),
Dr Marie Dziadek and Dr George Clark.
3.8
The Committee considered the application by teleconference on 6 December
2005.
4
Consideration
The Committee’s approach to the Consideration
4.1
In accordance with clause 24 of the Methodology, the approach to the
consideration adopted by the Committee was to first examine the scope and
purpose of the application, and the range of organisms applied for, then to
look sequentially at the identification, assessment and evaluation of risks,
costs and benefits. Those risks identified as significant were assessed (clause
12). Costs and benefits were assessed in accordance with clause 13 of the
Methodology. Qualitative scales used by the Committee to measure
likelihood and magnitude of risks, costs and benefits were provided in
Appendix 3 of the E&R Report.
4.2
Interposed with this was the consideration of the adequacy of the proposed
containment regime, and the ability of the organisms to escape and to
establish undesirable self-sustaining populations. Management techniques
were considered in relation to the identified risks. The Committee set
controls to satisfactorily provide for the matters in the Third Schedule (Part
I) of the Act and additional controls were considered in relation to any
residual risks that required further consideration.
4.3
Benefits associated with this application were considered in accordance with
clauses 9, 10, 13 and 14 of the Methodology and section 6(e) of the Act.
4.4
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. The approach follows
the decision path outlined in Appendix 1 of the E&R Report.
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Sufficiency of information
4.5
The Committee determined that the information supplied was relevant and
appropriate to the scale and significance of the risks, costs, and benefits
associated with the organism and was sufficient for consideration. The
comments received from DoC were noted by the Committee.
Purpose of the Application
4.6
The purpose of this application is to generate E. coli, and C. albicans, C.
dubliniensis, C. glabrata, C. krusei, C. tropicalis, S. cerevisiae, A.
fumigatus, and C. neoformans which have been genetically modified to alter
azole or echinocandin drug targets, or, genes involved in resistance to these
antifungal drugs. This will provide the applicant with tools to study the
interaction of antifungal drugs with their targets, and the mechanisms of
drug resistance in fungi.
4.7
In accordance with section 45(1)(a)(i) of the Act, the Committee determined
that this purpose falls within the scope of section 39(1)(a) of the Act: the
development of any new organism.
Scope of Application and Organism Description
4.8
The scope of the organisms subject to this approval is limited to the
following description:
4.9
Escherichia coli (non-pathogenic strains), Candida albicans, Candida
dubliniensis, Candida glabrata, Candida krusei, Candida tropicalis,
Saccharomyces cerevisiae (non-pathogenic strains), Aspergillus fumigatus
and Cryptococcus neoformans, genetically modified by plasmid vectors or
E. coli/yeast shuttle vectors containing selectable markers , reporter genes,
standard protein tags, regulatory sequences and donor DNA from the fungi
listed above. Donor DNA shall encode azole or echinocandin drug targets, or
genes involved in azole or echinocandin drug resistance.

4.10
Only one drug target or resistance mechanism will be investigated in
each modification.
The azoles act by inhibiting 14α –lanosterol demethylase, an enzyme
involved in sterol biosynthesis, whereas echinocandins inhibit glucan
synthase, an enzyme involved in the synthesis of fungal cell walls. To study
the interaction of antifungals with their drug targets, the expression of these
drug target genes will be modified by deletion, site-directed mutagenesis and
over-expression in the host fungi. The expression of genes involved in
resistance, such as drug efflux pumps, will be modified by deletion, sitedirected mutagenesis and over-expression. These types of modifications will
enable the applicant to examine how fungal cells develop resistance to the
azoles or echinocandin antifungal drugs.
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4.11
The proposed development of the genetically modified fungi in containment,
fall under schedule 1 (k) of the HSNO (Low Risk Genetic Modification)
Regulations (2003) developments that are not low-risk genetic
modifications. Schedule 1 (k): “developments involving modifications to
pathogenic micro-organisms that result in resistance to antibiotics used for
clinical or veterinary treatment of infections caused by that microorganism”.
4.12
The Committee noted that the genetically modified organisms would still
remain susceptible to at least two other classes of antifungal drugs used
clinically to treat fungal infections. The Committee also noted that none of
the antimicrobial drugs used for selection of the genetically modified fungi
are used to treat fungal infections in human medicine.
5
Containment
5.1
In assessing risks and costs, the Committee considered issues affecting the
adequacy of the containment regime (in accordance with section 45(1)(a) of
the Act); the potential for population establishment and population
eradication (section 37 of the Act and clauses 10(e) and 10(f) of the
Methodology); and other matters in order to give effect to the purpose of the
Act (section 45(2)(b)). Risk management techniques were used in relation to
the identified risks and costs (clauses 12(d) and 24 of the Methodology). As
such, the assessment of risks and costs (refer to section 6 of this decision)
was taken into account in setting the containment requirements that are
discussed in this section.
5.2
For the purposes of sections 37, 43 and 45(1)(iii) of the Act the Committee
has considered the biological characteristics of the organism, potential
pathways for escape of the organism from containment and the ability of the
organisms to escape containment and to establish a self-sustaining
population, and the ease with which such a population could be eradicated.
A set of controls to manage the risk of escape from containment are listed in
Appendix 1 of this Decision.
Ability to Escape from Containment
Biological characteristics of the organisms
5.3
The host organisms to be modified are microorganisms which fall into two
host categories. Non-pathogenic laboratory stains of E. coli and S. cerevisiae
are category 1 host organisms3 that are unlikely to cause disease in humans,
animals, plants or fungi. The E. coli and S. cerevisiae strains to be used in
the study will have nutritional requirements that make it highly unlikely that
these strains will be able to survive outside laboratory culture. The proposed
genetic modifications of these organisms will not increase the pathogenicity
of these hosts or increase their ability to escape from containment.
3
As defined in the HSNO (Low-Risk Genetic Modification) Regulations 2003.
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5.4
The other host organisms to be used as hosts for genetic modifications are
the following seven species of fungi: C. albicans, C. dubliniensis, C.
glabrata, C. krusei, C. tropicalis, A. fumigatus and C. neoformans. These
are category 2 hosts organisms, risk group 24, which may cause disease in
humans but are unlikely to be a serious hazard to laboratory personnel. The
biological characteristics of the host organisms have been described in
section 3 of the E&R report.
5.5
The Candida species are sensitive to desiccation and are normally dispersed
between mammals by contact. The route of infection by A. fumigatus and C.
neoformans is by inhalation of spores and aerosols.
5.6
Therefore the following containment regime is designed to contain fungal
cultures and spores, and bacterial cultures.
Containment regime
5.7
The controls imposed by this approval (as specified in Appendix 1) establish
a containment regime that addresses the matters detailed in the Third
Schedule Part I of the Act. Additional controls to give effect to the purpose
of the Act have also been included.
5.8
The host organisms to be genetically modified are microorganisms and
therefore the Committee imposes controls 1.1, 1.2, 1.3 and 1.4 (Appendix 1),
which require the genetically modified organisms to be developed and
maintained within a MAF registered containment facility subject to
construction, operational and management measures designed for the secure
containment of microorganisms. These controls impose the provisions of the
MAF/ERMA New Zealand Standard 154.03.02 Containment Facilities for
Microorganisms.
5.9
Having considered the risks associated with the development of these
microorganism, the Committee considers that Physical Containment level 2
(PC2) for microorganisms as defined in the AS/NZS 2243.3:2002 Standard
and modified by the MAF/ERMA Standard 154.03.02 Containment
Facilities for Microorganisms, is the appropriate containment level for the
opportunistic pathogenic fungi (category 2 host organisms), and Physical
Containment Level 1 (PC1) for E. coli and S. cerevisiae (category 1 host
organisms).
5.10
The Committee is satisfied that the genetic modifications are not anticipated
to affect the ability of the fungi to escape from containment. However, as A.
fumigatus produces spores, and the route of infection for A. fumigatus is
usually inhalation of spores, and for C. neoformans inhalation of aerosols,
the Committee has imposed an additional control to manage these risks. The
additional control requires that all manipulations with A. fumigatus and
procedures likely to generate aerosols containing C. neoformans, to be
carried out in a class II biological safety cabinet within the PC2 facility (as
4
As defined in the HSNO (Low-Risk Genetic Modification) Regulations 2003
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described by clause 1.3.5.2 of the AS/NZS 2243.2:2002) to prevent the
release of airborne spores to the environment (control 8.1, Appendix 1).
Potential pathways for escape from the containment facility
5.11
The Committee has identified the following potential routes by which the
organism may escape from containment:

Accidental release of the pathogen (or its spores) or any viable genetic
material

Escape of organisms as air-borne spores or aerosols

Escape in transit between facilities
5.12
The Committee considered that accidental release of the organisms may
occur by inadvertent removal by staff from the containment facility or
through removal by unauthorised persons gaining entry to the facility. The
presence of other organisms in the facility could pose another pathway of
escape. Other pathways of escape include incorrect handling (human error),
sabotage or natural disasters that breach the structural integrity of the
containment facility.
5.13
The inadvertent or deliberate removal of organisms from the containment
facility by unauthorised people (deliberate theft or sabotage) is considered to
be highly improbable because of the access restrictions (controls 2.1 and
4.1). The Committee considers that controls 1.2 and 7.1 are adequate to
ensure that staff training would reduce the likelihood of accidental release
from containment as a result of incorrect handling or procedure by personnel
to highly improbable.
5.14
The escape of genetically modified organisms can also be mediated by
rodents, birds or invertebrates. The Committee considers that this is highly
unlikely given the containment measures (Control 3.1). The MAF/ERMA
New Zealand Standard 154.03.02 also includes provisions for fitting fly
screens on windows that can be opened, if this is a potential pathway of
escape. The Committee considers that it is extremely unlikely that this will
occur.
5.15
The Committee identified the possibility that a breach of containment as a
result of natural disaster, could occur. In the event of an escape, the
Committee considers that the contingency plans should be implemented
immediately (control 5.2).
5.16
The Committee noted that unlike the other fungi, A. fumigatus produces
desiccation-resistant spores, and noted that the applicant sought advice on
working with A. fumigatus from Professor David Denning and Dr Michael
Anderson, Faculty of Life Sciences, University of Manchester, UK
(http://www.aspergillus.man.ac.uk). The applicant proposed to contain A.
fumigatus within tissue culture flasks rather than petri dishes, to reduce the
risk of escape of spores. Procedures with C. neoformans will be carried out,
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wherever possible, in sealed containers to minimise exposure of researchers
to aerosols. Fungal cultures will be disposed of by autoclaving in accordance
with section 4.6 (requirements for disposal of microorganisms and biological
waste) of the MAF/ERMA Standard 154.03.02.
5.17
To reduce the likelihood of escape of spores, all work requiring handling of
spores by laboratory personnel will be carried out within a class II biological
safety cabinet (as described by clause 1.3.5.2 of the AS/NZS 2243.2:2002).
Since UV light is not very effective in killing A. fumigatus spores,
disinfection of the biological safety cabinet will be achieved by a
bactericidal, virucidal, and fungicidal disinfectant such as Virkon.
5.18
The Committee considers that the use of a class II Biological Safety Cabinet
in these circumstances is appropriate, and has imposed an additional control
to this effect (control 8.1). The Committee considers that this control 8.1
together with the standard microbiology aseptic techniques required by the
AS/NZS 2243.3:2002 will reduce the likelihood of genetically modified
fungal spores or aerosols escaping from the facility to improbable.
5.19
The MAF/ERMA Standard 154.03.025 provides for transfers between
facilities of microorganisms in culture and sets minimum packaging
requirements6. The Committee considers that if all of these conditions are
met, then the likelihood of escape of fungi during transit will be improbable.
Control 1.4 would mandate these minimum conditions for any transfer of
approved microorganisms.
5.20
The Committee considered the above containment regime against each of the
matters specified in Schedule 3 Part 1 of the Act and endorse the analysis
presented in paragraphs 4.8 to 4.43 of the E&R Report. Having considered
the biological characteristics of the organism, the potential pathways by
which the organism could escape from containment and the containment
regime the Committee conclude that the likelihood of the organisms
escaping from containment is highly improbable.
Ability of Organism to Establish a Self-sustaining Population and the Ease of
Eradication
5.21
In accordance with sections 43 and 37 of the Act the Committee considered
the ability of the organism to establish self-sustaining populations, should it
escape from containment and the ease with which such populations could be
eradicated.
5.22
The Committee considered that the E. coli and S. cerevisiae strains (category
1 host organisms) are highly unlikely to establish self-sustaining populations
outside containment due to their nutritional requirements.
5
Sections 4.7, 4.8
6
Packaging Instructions No 650 of the IATA Dangerous Goods Regulations
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5.23
The Committee noted that the Candida species (category 2 host organisms)
are relatively sensitive to desiccation and are unlikely to survive out of
culture within the laboratory unless transferred directly to mammals. This
statement was supported by a recent publication supplied by the applicant.
The non Candida species however, C. neoformans and A. fumigatus are less
sensitive to desiccation and so may be able to establish a self-sustaining
population in the highly improbable event of an escape from containment.
5.24
The Committee noted that in the absence of selection pressure such as the
application of antifungal drugs, there will be no selective advantage for
genetically modified fungi resistant to azole or echinocandin drugs. The
Committee considered that the potential nutritional deficiencies, impaired
growth and increased metabolic burden caused by the genetic modifications
and the absence of selective advantage would reduce the likelihood of the
modified fungi surviving outside containment.
5.25
The Committee considered that it is highly improbable that genetically
modified fungi could establish self-sustaining populations outside
containment. The Committee considered that such a population could be
eradicated by antifungal treatment but noted that detection would be
unlikely.
6
Identification of Risks, Costs and Benefits
6.1
The Committee identified risks and costs related to the application in
accordance with clauses 9 and 10 of the Methodology, which incorporate
sections 5, 6, 8 and 43 of the Act.
6.2
The Committee identified no significant risks to the environment, public
health, Māori culture, or the economy, associated with developing
genetically modified E. coli and S. cerevisiae. Therefore, no further
assessment of the effects of these organisms has been conducted.
6.3
The other host organisms in this application; C. albicans, C. dubliniensis, C.
glabrata, C. krusei, C. tropicalis, A. fumigatus, and C. neoformans are all
opportunistic pathogens. These organisms are capable of causing minor
mucosal and skin infections in humans, and serious disseminated disease in
severely immunocompromised individuals. Some of the proposed genetic
modifications could increase the resistance of the fungi to the azole or
echinocandin classes of the antifungal drugs currently used to treat such
infections.
6.4
Based on the biology of these organisms and the proposed genetic
modifications, the Committee has identified the following potential adverse
effects which may occur if the genetically modified fungi escape from
containment and established a self sustaining population.
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Potential adverse effects on the environment:
6.5
The Committee identified the following as significant potential adverse
environmental effects (assessed below in section 7 of this decision).

Potential for displacement of other fungi in the same ecological niche;

Potential for spread of resistance to azole or echinocandin drugs to
naturally occurring fungi.
Potential adverse effects on public health:
6.6
The Committee identified the potential for genetically modified fungi to
infect research personnel working with the fungi (assessed below in section
7 of this decision).
Potential adverse effects on Māori culture:
6.7
The Committee identified the following as significant potential adverse
effects on Māori culture (assessed below in section 7 of this decision).

Potential to affect the well being of the mauri of individuals working in
the laboratory;

Potential for adverse effects on tāonga species.
Potential adverse effects on social or the community:
6.8
The Committee identified no potential adverse effects on society or the
community, associated with developing genetically modified fungi.
Therefore, no further assessment of these potential effects has been
conducted.
Potential adverse effects on the economy:
6.9
The Committee identified no potential adverse effects on the market
economy, associated with developing genetically modified fungi. Therefore,
no further assessment of these potential effects has been conducted.
Potential effects on New Zealand’s international obligations:
6.10
The Committee noted that New Zealand’s obligations under the Cartegena
Protocol are satisfied by the requirements of the HSNO Act and ERMA New
Zealand’s undertaking to deposit information concerning all Living
Modified Organisms (LMOs) developed in New Zealand onto the Biosafety
Clearing House web site.
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Identification of potential beneficial effects
6.11
The Committee has identified the following potential direct benefits
associated with this application (assessed below in section 7 of this
decision).

7
Gains in scientific knowledge about the interaction of antifungal drugs
and their targets, and mechanisms of drug resistance in fungi, through
the development of a suite of tools for research.
Identification, Assessment and Evaluation of Risks,
Costs and Benefits
7.1
In section 6 above, the Committee identified potentially significant adverse
and beneficial effects of the fungi. These are assessed below.
Potential for displacement of other fungi in the same ecological niche
7.2
The Committee considered all the potential outcomes of the genetic
modification of fungal hosts as outlined in paragraph 5.18 of the E&R
Report. By deleting genes involved in biosynthetic pathways, genetically
modified fungi will require growth media supplemented with additional
nutrient. In addition, the range of proposed genetic modifications such as the
alteration of drug targets could introduce a fitness cost on fungal cells as
some of these genes have essential functions within the cell. The overexpression of genes could also place an increased metabolic burden on cell
growth. To displace wild type fungi from their ecologically niches, the
genetically modified fungi must have some selective advantage over their
wild type counterparts. In the absence of any selection pressure such as the
application of antifungal drugs, the Committee considers the likelihood of
this occurring to be reduced.
7.3
The Committee considers the likelihood of these modified fungi escaping
from containment and displacing wild type fungi to be highly improbable,
and would be of minimal effect. Therefore, the risk of adverse effects to
naturally occurring fungi is considered to be negligible. The Committee
considers that this risk can be adequately managed by the proposed
containment measures.
Potential for the spread of resistance to azole or echinocandin drugs from
genetically modified fungi to naturally occurring fungi
7.4
The Committee considered the potential for genetically modified fungi to
transfer the resistance trait to naturally occurring fungal communities and
thereby altering natural biodiversity. The Committee considers that for this
to occur, there has to be both genetic mechanisms to transmit modified
genetic material conferring resistance to antifungal drugs between fungal
populations, and selection pressure in the environment. For reasons outlined
in paragraphs 5.25 – 5.29 of the E&R Report, the Committee considers the
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likelihood of antifungal drug resistance genes spreading between populations
of fungi is highly improbable.
7.5
In the highly improbable event that transmission of resistance occurred, the
Committee considered that the spread of resistance to azole and
echinocandin drugs amongst naturally occurring fungi would be localised
and could be contained if the location is known. The Committee considered
that the magnitude of such an effect would be minimal to minor, and
assessed this risk as negligible.
Potential for infection of research personnel
7.6
The fungi C. albicans, C. dubliniensis, C. glabrata, C. krusei, C. tropicalis,
A. fumigatus, and C. neoformans are all opportunistic pathogens which can
colonise humans and can potentially cause infection. The Committee
considered the potential for the genetically modified fungi to infect research
personnel working with the fungi and noted that this potential adverse effect
would be more severe in immunocompromised individuals. Severely
immunocompromised individuals are most likely to be susceptible and
normally these individuals require hospitalization. Therefore, the Committee
considered the likelihood of exposure of susceptible individuals to this
potential adverse effect is highly improbable.
7.7
The Committee considered that within containment, direct contact of
researchers with genetically modified fungi could occur if PC2 operating
procedures were not adhered to. The Committee noted that this is most likely
to occur through a breach of containment, or inappropriate handling
procedures.
7.8
The Committee considered that PC2 standard operating procedures will
minimise the risk of colonisation of researchers by the Candida species, as
outlined in paragraphs 5.35-5.36 of the E&R Report. The risk of infection by
A. fumigatus and C. neoformans, which gain access to the host by inhalation,
will be reduced by additional operational containment practices (the use of a
class II Biological Safety Cabinet).
7.9
The Committee also considered the transfer of genetically modified fungi to
other individuals outside of containment by contact with a contaminated
laboratory worker. The Committee noted that there is no evidence for
efficient transfer of C. albicans strains between healthy individuals, and
even if inoculation was to occur, the probability that the modified fungi
could replace endogenous strains and colonise the healthy human is low. On
the basis of the analysis in paragraphs 5.38-5.45 of the E&R Report, the
Committee considered that the likelihood of infection of laboratory workers
or other persons with the genetically modified fungi is highly improbable,
and the magnitude of the risk of potential infection by the fungi ranges from
minimal to minor. The Committee has assessed the risk of infection of
research personnel or other persons by these modified fungi to be negligible.
7.10
The Committee noted that in paragraph 6.3 of the E&R Report (Previous
similar applications) an additional control was imposed on an approval
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(GMO04/UO005) requiring “Persons known to be immunocompromised
shall not work with genetically modified C. albicans”. This matter was
addressed in paragraphs 5.47 – 5.48 of the E&R Report and considered to be
unnecessary as severely immunocompromised patients, who are at higher
risk of infection, are effectively prevented from exposure by being
hospitalised.
7.11
The applicant stated in an email submission that he wanted this matter
brought to the attention of the Authority. In at least four previous decisions
by IBSCs, two from Otago University and another two from Massey
University, no such additional controls were added. Although the control for
GMD04050 was added by the Otago IBSC, the IBSC now think that it is
impracticable as the term “immunocompromised” is not defined and it
would be impracticable to test the immune status of people working with the
genetically modified organisms (GMOs). In addition, it would raise difficult
privacy issues. Based on the analysis in the E&R Report and information
provided by the applicant, the Committee considered that the additional
control “Persons known to be immunocompromised shall not work with
genetically modified C. albicans”, was unnecessary.
Potential adverse effects on Māori culture
7.12
The Committee considered the potential Māori cultural effects of this
application in accordance with sections 6(d) and 8 of the HSNO Act 1996,
and the assessment framework contained in the ERMA New Zealand User
Guide “Working with Māori under the HSNO Act 1996”.
7.13
The Committee noted that in accordance with ERMA New Zealand policy
and guidelines, the applicant was not required to consult with Māori
regarding this application as it is a containment application that does not
involve genetic material from native flora and fauna or humans.
Nevertheless the applicant had provided information that demonstrates that
in similar circumstances with regard to research of this nature, the local Iwi
had no concerns.
Potential to affect the well being of the mauri of individuals working in the
laboratory
7.14
The area of potential adverse effect is to the well-being of the mauri of the
individual working within the confines of the laboratory environment, or in
the highly improbable event of organism escape, of other persons coming
into contact with the genetically modified fungi. The Committee considered
that the likelihood of this effect is highly improbable and, if exposure does
occur, the magnitude of this effect is minimal to minor (refer paragraphs
5.32 – 5.46 of E&R Report on public health). Therefore the Committee
considered that the risk of potential adverse effects on the well-being of the
mauri is negligible.
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Potential for adverse effects on taonga species
7.15
The Committee considered the potential for modified fungi to escape and
establish self sustaining populations, thereby causing adverse effects on
taonga species. However, there is no evidence that these genetically
modified fungi have any selective advantage over naturally occurring nongenetically modified fungi and therefore the potential for this application to
have adverse effects on taonga species is considered negligible.
Assessment of potential beneficial effects
7.16
The Committee identified the following potential direct benefits associated
with the application, in accordance with clauses 9, 10, 13 and 14 of the
Methodology, and section 6(e) of the Act:

7.17
Gains in scientific knowledge about the interaction of antifungal
drugs and their targets, and mechanisms of drug resistance in fungi,
through the development of a suite of tools for research.
The applicant states that gain in scientific knowledge could lead to the
following:

Publications in international peer-reviewed journals and conference
presentations which would enhance their reputation internationally
and strengthen their ability to attract future research funding;

Design of antifungal drugs that may be more effective in treatment of
patients with fungal disease;

The knowledge of the efflux-pump mediated drug resistance in fungi
may contribute to investigation of drug resistant tumours in cancer
chemotherapy.
7.18
The Committee considered that the primary benefit associated with the
development in containment of genetically modified fungi and Eschericia
coli, is increased scientific knowledge about how antifungal drugs interact
with their targets and how fungal cells become resistant to drugs. This
knowledge may have further clinical applications for the treatment of fungal
diseases and the design of new drugs. The Committee noted that as these
benefits are research based, the likelihood and magnitude of the potential
medical outcomes cannot be defined with absolute certainty.
7.19
However, the Committee considered that the primary benefit, a gain in
scientific knowledge, is very likely to be realised, and would be of
moderate value. This potential benefit is considered significant by the
Committee.
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8
Establishment of the approach to risk in the light of risk
characteristics
8.1
Clause 33 of the Methodology requires the Authority to have regard for the
extent to which a specified set of risk characteristics exist when considering
applications. This provision provides a route for determining how cautious
or risk averse the Authority should be in weighing up risks and costs against
benefits. In the present application clause 33 is influenced by the application
being “in containment” and the conclusion that the containment provisions
and other controls reduce biological and physical risks to a low level.
8.2
In relation to the biological and physical risks considered the containment
measures limit the extent to which exposure to the risks is involuntary. The
Committee also considers that there are no significant risks which are not
known or understood by the general public. It is considered that the
potentially significant risks are dependant upon the ability of the organisms
to cause disease in humans and that these risks are localised, controllable
and reversible. Given the Committee's finding that escape from containment
and population establishment is highly improbable, the extent to which these
risk characteristics are present does not warrant caution additional to that
required by section 7 of the Act.
9
Overall Evaluation of Risks, Costs and Benefits and of
the Adequacy of Containment
9.1
In evaluating the potential beneficial effects associated with this application
the Committee concludes that the key benefit of the proposed research is the
ability to contribute to scientific knowledge of the interactions of antifungal
drugs with their targets, and how resistance to antifungal drugs arise in
fungi. This was assessed as very likely to eventuate and to be of moderate
value.
9.2
The Committee noted that, with one minor exception, the proposed
modifications meet the criteria for a low-risk genetic modification
(prescribed in the HSNO (Low-Risk Genetic Modification) Regulations
2003). The minor exception is that the proposed developments may result in
genetically modified pathogenic fungi with increased resistance to either the
azoles or echinocandins classes of antifungal drugs; a risk assessed as
negligible. While some of the modifications may result in fungal strains with
an increased resistance to either the azoles or echinocandins, all genetically
modified fungi will remain susceptible to at least two other classes of
clinically used antifungal drugs. The proposed modifications will not
increase the host organisms’ pathogenicity, infectivity, virulence or ability to
escape from containment. Notwithstanding this single exception to the
criteria for a low-risk genetic modification, the Committee has undertaken a
full risk assessment as required by section 45 of the Act. All risks were
considered to be negligible.
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9.3
10
Having considered the biological characteristics of the organism, the
potential pathways by which the organism could escape from containment
and the containment regime the Committee conclude that the likelihood of
the organism escaping from containment is highly improbable. In the event
of an escape, the Committee considered that it is highly improbable that
genetically modified fungi could establish self-sustaining populations
outside containment. The Committee considered that such a population
could be eradicated by antifungal treatment but noted that detection would
be unlikely. In relation to the level of risk associated with the proposed
genetic modification, the containment requirements, including the additional
controls imposed by the Committee, were considered to be adequate.
Decision
10.1
In reaching its decision on this application, the Committee records that the
following criteria in the Act and the Methodology have been particularly
relied on (in accordance with clauses 21 and 36(2)(b) of the Methodology):
10.2
The application has been considered in the context of the purpose and
principles of the Act (sections 4-8 inclusive).
10.3
Pursuant to section 45(1)(a)(i) of the Act, the Committee is satisfied that the
purpose of the application falls under section 39(1)(a): the development of
any genetically modified organism.
10.4
In accordance with sections 45(1) and (4) of the Act, and clauses 9, 10 and
12 of the Methodology, the Committee concluded that each of the risks and
costs was negligible after taking account of the organism description and the
impact of containment and other controls set out in Appendix 1. Thus, the
Committee considered the application under clause 26 of the Methodology.
10.5
It is evident to the Committee that the potential benefits associated with the
development of this organism outweigh the costs.
10.6
The Committee is satisfied that the genetically modified fungi can be
adequately contained by the controls specified in this decision (section
45(1)(a)(iii)).
10.7
In accordance with clause 36(2)(b) of the Methodology, the Committee
records that in reaching this conclusion, it has applied the balancing tests in
section 45 of the Act.
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10.8
The application to develop in containment:
Escherichia coli (non-pathogenic strains), Candida albicans, Candida
dubliniensis, Candida glabrata, Candida krusei, Candida tropicalis,
Saccharomyces cerevisiae (non-pathogenic strains), Aspergillus fumigatus
and Cryptococcus neoformans, genetically modified by plasmid vectors or
E. coli/yeast shuttle vectors containing selectable markers7, reporter genes,
standard protein tags, regulatory sequences8 and donor DNA from the fungi
listed above. Donor DNA shall encode azole or echinocandin drug targets,
or genes involved in azole or echinocandin drug resistance.

Only one drug target or resistance mechanism will be investigated in
each modification.
is approved with controls in accordance with section 45(1)(a) of the Act.
As required under section 45(2) the approval is subject to the controls listed
in Appendix 1 of this decision.
Dr Kieran Elborough
Date
Chair, GMO Standing Committee of the Authority
Approval codes: GMD004136 – 44
BCH numbers: 11445 - 53
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
7
Including antibiotic resistance genes and genes involved in nutrition biosynthesis to select for
genetically modified fungi.
8
Fungal genes will be expressed under the control of their own promoters or alternative fungal
promoters that give high or low levels of expression.
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Appendix 1: Controls Required by this Approval
In order to satisfactorily address the matters detailed in the Third Schedule Part
I: Matters to be Addressed by Containment Controls for Development and Field
Testing of Genetically Modified Organisms9of the Act, and other matters in
order to give effect to the purpose of the Act (section 45(2)), the approved
organisms are subject to the following controls:
Containment Controls
1
To limit the likelihood of any accidental release of any organism or
any viable genetic material10:
1.1
The approved organisms shall be developed in, and maintained within a
containment facility which complies with these controls.
1.2
The person responsible for a particular research area and/or the person
responsible for the operation of the containment facilities (‘the facility’) shall
inform all personnel involved in the handling of the organisms of the
Authority’s controls.
1.3
The containment facility shall be approved by Ministry of Agriculture and
Forestry (MAF), in accordance with section 39 of the Biosecurity Act and the
MAF Biosecurity Authority/ERMA New Zealand Standard 154.03.0211:
Containment Facilities for Microorganisms.
1.4
The construction, operation, and management of the microorganism
containment facility shall be in accordance with the:
a)
Ministry of Agriculture and Forestry (MAF)/ERMA New Zealand Standard
154.03.0211: Containment Facilities for Microorganisms.
b) Australian New Zealand Standard AS/NZS 2243:3 200211 Safety in
Laboratories: Part 3: (Microbiological aspects and containment facilities),
excluding those deviations specified in section 4.2 of Standard 154.03.02.
c)
Physical Containment Level 1 (PC1) requirements of the above Standards
for E. coli and S. cerevisiae and Physical Containment Level 2 (PC2)
9
Bold headings refer to matters to be addressed by containment controls for new organisms
excluding genetically modified organisms, specified in the Third Schedule (Part II) of the
HSNO Act 1996.
10
Viable genetic material is biological material that can be resuscitated to grow into tissues or
organisms. It can be defined to mean biological material capable of growth even though
resuscitation procedures may be required, e.g. when organisms or parts thereof are sub lethally
damaged by being frozen, dried, heated, or affected by chemical.
11
Any reference to this standard in these controls refers to any subsequent version approved or
endorsed by ERMA New Zealand
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requirements of the above Standards for C. albicans, C. dubliniensis,
C. glabrata, C. krusei, C. tropicalis, A. fumigatus, and C. neoformans.
2
To exclude unauthorised people from the facility:
2.1
The identification of entrances, numbers of and access to entrances, and
security requirements for the entrances and the facility shall be in compliance
with the requirements of the standards listed in control 1.3 and 1.4 of this
document.
3
To exclude other organisms from the facility and to control
undesirable and unwanted organisms within the facility:
3.1
The exclusion of other organisms from the facility and the control of
undesirable and unwanted organisms within the facility shall be in compliance
with the standards listed in control 1.3 and 1.4 of this document.
4
To prevent unintended release of the organism by experimenters
working with the organism:
4.1
The prevention of unintended release of the organisms by experimenters
working with the organisms shall be in compliance with the standards listed in
control 1.3 and 1.4 of this document.
5
To control the effects of any accidental release or escape of an
organism:
5.1
Control of the effects of any accidental release or escape of the organisms shall
be in compliance with the standards listed in control 1.3 and 1.4 of this
document.
5.2
In the event of any breach of containment the contingency plan for the
attempted retrieval or destruction of any viable material of the organisms that
have escaped shall be implemented immediately. The contingency plan shall be
included in the containment manual in accordance with the Standards.
5.3
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.
6
Inspection and monitoring requirements for containment facilities:
6.1
The inspection and monitoring requirements for containment facilities shall be
in compliance with the standards listed in control 1.3 and 1.4 of this document.
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7
Qualifications required of the persons responsible for
implementing those controls:
7.1
The training of personnel working in the facility shall be in compliance with the
standards listed in control 1.3 and 1.4 of this document.
8
Controls additional to the requirements of Standard 154.03.02
(Matter 1)
8.1
All manipulations with Aspergillus fumigatus and procedures likely to generate
aerosols containing Cryptococcus neoformans will be carried out in a class II
biological safety cabinet within the PC2 facility.
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