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CSIRO PUBLISHING

Wildlife Research, 2007, 34, 578585 www.publish.csiro.au/journals/wr

Pest or prized possession? Genetically modified biocontrol


from an international perspective

Wendy R. Henderson A,C and Elaine C. Murphy A,B


A
Invasive Animals Cooperative Research Centre, University of Canberra, ACT 2601, Australia.
B
Department of Conservation, PO Box 13049, Christchurch, New Zealand.
C
Corresponding author. Email: wendy.henderson@invasiveanimals.com

Abstract. This article provides an overview of current research, regulations and international issues concerning
genetically modified (GM) organisms for use as biological controls of vertebrates. There is increasing interest in using
biotechnology to solve vertebrate pest problems around the world. A major issue lies in the fact that individual countries
focusing on internal problems of pest management may overlook the potential of transborder entry. Animals considered a
pest in one country may well be prized possessions in another, and research and management strategies should consider
the adverse effects of biocontrol agents entering the wrong country. There is a wealth of guidance in the form of national
and international regulations and ethics guidelines. However, current legislation and agreements may not be adequate to
ensure that all risks of GM biocontrols, particularly disseminating agents, have been considered from an international
perspective. Major issues include concerns of transboundary movement, non-target effects and the need for an inter-
national body to consult with and regulate the use of GM biocontrols. We live in a finite and interconnected world: it is
vital that impacts of potential control strategies are assessed at a local and international level, and from social, environ-
mental and economic perspectives.

Introduction
The introduction of invasive plants, animals and micro- a species in another country where it is considered a pest (Gilna
organisms has caused huge economic and environmental losses et al. 2005; Williams 2007). Similarly, concern has been raised
in many countries (Pimentel et al. 2001; Seamark 2001). over the risks of a GMO designed to preserve a species in one
Invasive animals, such as rats (Rattus spp.) and feral cats (Felis country compromising pest-control programs in another
catus) have been shown to be directly responsible for the extinc- (Angulo and Cooke 2002; Anon. 2004).
tion of native species. Without control or eradication programs Regulations governing the research and release of such GM
they will continue to cause further declines and extinctions biocontrol agents differ considerably in content and stringency
(Atkinson 1989; Pimentel et al. 2001; Courchamp et al. 2003). from country to country. Various levels of regulations and
The development of biotechnology, including advances in guidelines also exist, from individual institutions to federal
genetic manipulation techniques, may provide new opportuni- governments and international agreements. There is currently
ties for vertebrate pest control. Controlling reproduction by vec- some confusion about the relevance and authority of some of
tored immunocontraception is one such technique (Barlow these instruments, and what steps researchers are required to
2000; Seamark 2001). Biotechnology approaches may also take during the development of GM biocontrols. There is
provide answers to problems associated with non-target effects growing support for an international authority to deal with these
and innate or developed resistance to current controls (Seamark issues (CBD 2004). The purpose of this article is to outline the
2001). Research into genetically modified (GM) biological con- current status of the research, regulations and international
trols for vertebrate populations has been undertaken in several issues regarding GM biocontrols for vertebrate populations.
countries, including Australia, New Zealand, Spain and the Some suggestions from an international perspective for a way
United States (Torres et al. 2001; Kapuscinski and Patronski forward are also included.
2005; Cowan et al. 2006; Hardy et al. 2006; Angulo and
Brcena 2007; Redwood et al. 2007; Strive et al. 2007; van Research and releases of GM biocontrols
Leeuwen and Kerr 2007). Several research projects have investigated GM biocontrol
From an international perspective, concerns have been raised agents to decrease introduced vertebrate pest populations, or
of transborder entry of GM biocontrol agents (Gilna et al. 2005; preserve valued native species (see Table 1). The Australian
Hill and Sendashonga 2006). These concerns highlight the sig- research on GMOs for fox (Vulpes vulpes), rabbit (Oryctolagus
nificant problem of pest control we face as an international cuniculus) and house mouse (Mus musculus domesticus) control
community: what is considered a pest in one country may be a has ceased owing to technical obstacles (Redwood et al. 2007;
prized possession in another. There are concerns that harm to Strive et al. 2007; van Leeuwen and Kerr 2007). However,
valued native species may be caused by the introduction of a advanced research continues on transmissible biocontrol agents
GM organism (GMO) originally designed to eradicate or reduce for possum (Trichosurus vulpecula) population reduction in

CSIRO 2007 10.1071/WR07062 1035-3712/07/070578


Genetically modified biocontrol Wildlife Research 579

New Zealand and rabbit preservation in Spain. The Spanish GM omatosis/RHD vaccine, for example). Most objections would
virus (conferring immunity to myxomatosis and rabbit haemor- probably involve human health issues (such as reactions to vac-
rhagic disease) has been released on a small scale, in an island cinia strains; for example, see Omlin 1997; Rupprecht et al.
field test (Torres et al. 2001). 2001) and political differences.
The GM rabies vaccine (designated VRG) was the first GMO
for vertebrate populations to be released into environmentally International instruments
widespread use 20 years ago. It is interesting to speculate The main international-level instruments (encompassing a
whether this vaccine would be released in the current regulatory range of agencies and agreements) involved in the development
and political climate. The threat of uncontrolled rabies would be and release of GM biocontrol agents are: the Convention on
a major factor in its acceptance, but there is still a mixed reaction Biological Diversity (CBD), the International Plant Protection
to the use of GMOs per se. While a full risk assessment of VRG Convention (IPPC), the OIE, and the Sanitary and Phytosanitary
is beyond the scope of this article, it seems probable that release Agreement (SPS Agreement) of the World Trade Organisation.
would still be approved in at least some countries comfortable A brief review of the current status of relevant principles is pro-
with using GMOs. Many safety aspects of VRG were examined vided in Table 2.
over 10 years of laboratory studies and strictly controlled field There are many other international and regional instruments
releases. Factors considered at the time include: host virus char- relevant to managing the spread and adverse effects of invasive
acteristics, target specificity of virus and baits, field durability, alien species. Alliances exist between some of these and the
pathogenicity, transmissibility, possible recombination with major instruments described in Table 2, recognising overlapping
other viruses, and consideration of sites prior to release (Hanlon roles and aiming to reinforce information exchange and
et al. 1989; Boulanger et al. 1996; Pastoret and Brochier 1996; improve coordination (e.g. CBD 2006).
Cliquet and Aubert 2003). As a non-transmissible agent, VRG The only international instrument specifically dealing with
would be likely to receive higher support for release than a dis- transborder movement of GMOs for use with animals is the
seminating agent: both the World Organisation for Animal Cartagena Protocol on Biosafety, which states that participating
Health (OIE) and World Health Organisation have expressed parties must ensure that any GMO (including developmental
their concern at using disseminating agents for wildlife man- phases) is used in a way that prevents or reduces the risks to bio-
agement (see Hinds et al. 2000). In addition, from a transborder logical diversity. As of August 2007, 142 countries have
perspective, VRG is unlikely to cause problems with opposing accepted to be bound by this treaty. Australia and the United
wildlife-management strategies (in contrast to the Spanish myx- States are notable exceptions.

Table 1. Recent or proposed research into GM biocontrols

Area and species GM organism References

Australia
European carp Daughterless technology transgenic approach blocking the expression of the Thresher and Bax (2003)
(Cyprinus carpio) female-determining aromatase gene
Cane toad (Bufo marinus) Daughterless technology transgenic approach; and potentially transmissible Molloy and Henderson (2006)
viral-vectored delivery of gene to prevent metamorphosis
Wild house mouse (Mus Mouse cytomegalovirus expressing mouse zona pellucida (ZP) subunit 3 gene, Redwood et al. (2007),
musculus domesticus) as transmissible agent to prevent fertilisation Williams et al. (2007)
Rabbit (Oryctolagus Myxoma virus expressing ZP glycoproteins, as transmissible agent to prevent Hamilton et al. (2005),
cuniculus) fertilisation; and trypanosome Trypanosoma nabiasi as transmissible Hardy et al. (2006),
immunocontraceptive agent (suggested research) van Leeuwen and Kerr (2007)
Fox (Vulpes vulpes) Vaccinia virus, canine herpesvirus and Salmonella typhimurium, expressing Bradley et al. (1997), Reubel et al. (2005),
various gamete proteins, to prevent fertilisation Hardy et al. (2006), Strive et al. (2006,
2007)
New Zealand
Australian brushtail possum Possum-specific nematode (Parastrongyloides trichosuri), to produce Cowan et al. (2006), Grant et al. (2006a,
(Trichosurus vulpecula) transmissible sterilising agent 2006b), Newton-Howes et al. (2006)
Spain
Rabbit (Oryctolagus Myxoma virus expressing rabbit hemorrhagic disease (RHD) virus capsid Brcena et al. (2000), Torres et al. (2000,
uniculus) protein, to confer transmissible immunity against myxomatosis and RHD 2001), Angulo and Brcena (2007)
United States
Exotic fish Triploid or transgenic fish to produce sterile individuals or to disrupt Kapuscinski and Patronski (2005)
development
United States and Europe
Raccoon (Procyon lotor), Vaccinia virus expressing rabies glycoprotein for non-transmissible Brochier et al. (1996), Pastoret et al.
coyote (Canis latrans), rabies vaccination (1993), Hanlon et al. (1998)
skunk (Mephitis mephitis)
and fox (Vulpes vulpes))
580 Wildlife Research W. R. Henderson and E. C. Murphy

The application of modern international law, as contained in National regulation of GM biocontrols


international instruments, varies from country to country, Different systems of regulation are currently being used by key
depending on their domestic law. For some, a signed treaty is countries to control research and environmental release of GM
binding upon their courts. However, for most countries it is per- biocontrol agents. These are briefly outlined in Table 3. In most
suasive only; even if they have signed an international agree- countries the regulation is split between several agencies and over
ment, that agreement still needs to be ratified by the domestic multiple acts of legislation. The degree of enforceability of the
legislature and, in many cases (as in Australia, New Zealand, the legislation and guidelines also varies considerably from country
United States and the European Union) incorporated into a to country, particularly with regard to international consultation.
domestic statute before being truly enforceable.
In the case of the Cartagena Protocol, non-compliance is International conferences, issues and implications
dealt with through a Compliance Committee, and may involve: International issues concerning the use of disseminating GM
(a) providing financial and technical assistance to facilitate biocontrol agents centre on the rights, objectives and ability of
compliance; (b) issuing a caution to the concerned party; individual countries to manage pests or endangered species,
(c) publishing cases of non-compliance in the Biosafety global responsibilities to preserve trading arrangements and the
Clearing-House; and (d) taking other measures such as suspen- environment, and transparency and availability of information
sion of trade, and/or suspension of some rights or privileges and consultation.
accorded to the Party (CBD 2007). To date, there have been no A symposium on Rabbits and Rabbit Haemorrhagic
reported cases of non-compliance. Disease (RHD): Disseminating Genetically Modified Organ-

Table 2. International instruments relevant to GM biocontrols

Agency Principle Brief description Reference

Convention on Article 3 Each party has sovereign right to exploit their own resources, but with due United Nations (1993)
Biological regard to the environment of areas beyond their national jurisdiction.
Diversity (CBD) Article 5 Parties should cooperate with each other on biodiversity matters of mutual interest. United Nations (1993)
Article 8 Risks associated with the use and release of GMOs that could have adverse United Nations (1993)
environmental effects should be managed by contracting parties.
Article 14 Requires environmental impact assessments, including consideration of areas United Nations (1993)
beyond national jurisdiction. Also states issue of liability and redress should
be examined.
Decision V1/23 Deals with intentional and unintentional introduction of species and mitigation CBD (2002)
of impacts of invasive alien species.
CBD Cartagena Advanced Preimport information provided before release of GMO. CBD (2000)
Protocol on Informed
Biosafety Agreement
Article 16 Risk assessments should consider international implications; transboundary CBD (2000)
movement should be prevented.
Article 17 Parties should take necessary steps in the event of an accidental release. CBD (2000)
International Plant International Concerned with the import and release of self-replicating plant biocontrol IPPC Secretariat (2005a)
Protection Standards for agents, including consideration of risk to other countries.
Convention Phytosanitary
(IPPC) Measure 3
(ISPM 3)
ISPM 11 Includes plant pest risks posed by GMOs, requiring the risks of the GMO itself IPPC Secretariat (2005b)
to be analysed and the consequences of its genetic material moving to
another organism.
World Trade Sanitary and Trade-related rules concerning sanitary and phytosanitary measures: countries WTO (1995)
Organisation Phytosanitary retain their right to ensure that the food, animal and plant products they import
(WTO) Agreement (SPS are safe, but should not use unnecessarily stringent measures as barriers to trade.
Agreement)
World Organisation Terrestrial Animal International coordination and cooperation on infectious animal diseases, with OIE (2006)
for Animal Health Code regard to trade in terrestrial animals, their genetic material and products.
Health (OIE)A (recognised in
SPS Agreement)
Aquatic Animal International coordination and cooperation on infectious animal diseases, with OIE (2007)
Health Code regard to trade in aquatic animals, their genetic material and products.
(recognised in
SPS Agreement)
A
OIE is considering GMO issues (OIE 2005): has concurred that disseminating agents are not desirable but that GMO research should proceed with extreme
caution (Anon 2004).
Genetically modified biocontrol Wildlife Research 581

isms (GMOs) and Conflicting International Objectives was GMOs. The main points from the discussion threads are sum-
held at the 3rd International Wildlife Management Congress marised in Table 4.
(5 December 2003, Christchurch, New Zealand) (Anon. 2004). A few participants questioned whether this type of bio-
This meeting highlighted the concerns of research projects on control research should continue at all in the current precau-
rabbit biocontrol with opposing management strategies (i.e. tionary climate, particularly with regard to its cost and possible
eradication of rabbits in Australia versus rabbit preservation in pressure by shareholders to release GMOs. However, there was
Spain) (Anon. 2004). The discussion was extended to include general consensus that the research should continue, at least in
the GM biocontrol of possums in New Zealand and house mice the biosecure laboratory phase: to explore options for biocontrol
in Australia. The potential for these biocontrol agents to enter and perhaps come up with unforseen answers to the problems
the wrong country and cause unwanted effects on the same (or discussed.
related) species was acknowledged (Anon. 2004). It was noted Participants agreed that there was a need for a consistent
that there has been little consideration of this risk to date, or of framework for international debate, with set standards to make
the risk of transfer of genetic material from such a GMO to it clear who is responsible for what, and to what degree. The
another wild-type organism (Anon. 2004). However, since that process could be guided by existing protocols in the Cartagena
time, an assessment has been made of the risks of inadvertent Protocol and other CBD and OIE codes. Serious difficulties in
export of the GM immunocontraceptive virus for house mice assessing risks and reaching consensus on a decision to release
from Australia (Williams 2007). GM biocontrol agents could arise from different countries atti-
Symposium participants explored whether current regula- tudes to the pest or the GMO, perceived national rights and inad-
tions are adequate to manage the risks associated with the equate infrastructure capacity. It was pointed out that a database
development and deployment of disseminating GMOs. It was of experience could be built up to assist decisions on risk and
concluded that they are not adequate, being limited by rights of release this has occurred with other agencies such as the
sovereignty and providing guidance only (Anon. 2004). It was Office of the Gene Technology Regulator in Australia.
agreed that research should proceed with caution, and that an Participants concluded that it is difficult with our current
international consultation process could help manage the risks state of knowledge to guarantee that the use of GMOs can be
and problems inherent in using GMOs in animal population limited to any particular region; therefore, the release of GMOs
management (Anon. 2004). should be everybodys concern. Currently, it is up to researchers
An online conference on Biosafety Considerations in the to inform regulatory and ethics-based agencies of progress, and
Use of Genetically Modified Organisms for Management of it is up to those agencies to consult with each other to approve
Animal Populations was hosted by the CBD from 18 October to or disallow GMO releases. It was generally concluded that a
15 November 2004 (CBD 2004). A total of 495 participants clearer-cut and more consistent process of consultation and
registered for the conference from 104 countries (Galloway management is needed, and that both national and international
Maclean 2005). The emphasis of discussions was on inter- mechanisms need to be strengthened to adequately deal with
national consultation processes for development and release of disseminating GM biocontrol agents. It was also acknowledged

Table 3. National regulatory instruments relevant to GM biocontrols

Country Primary legislation/guidelines Level of enforcement

Australia Gene Technology Act 2000, Biological Control Act 1984, Australia New National Acts enforceable. No legal requirement to consult
Zealand Food Authority Act 1991, Therapeutic Goods Act 1989, internationally before approving GMO releases (i.e.
Agricultural and Veterinary Chemicals Code Act 1994, Industrial Ethical Framework not enforceable by law). International
Chemicals [Notification and Assessment] Act 1989, and various state law considerations of CBD (but not Cartagena Protocol).
and territory regulatory schemes, Framework for the Development of
Ethical Principles in Gene Technology (GTEC 2006).
New Zealand Hazardous Substances and New Organisms (HSNO) Act 1996, National Acts enforceable. Section 6 of the HSNO Act
Biosecurity Act 1993, Agricultural Compounds and Veterinary requires consideration of international obligations.
Medicines Act 1997, Food Act 1981, Medicines Act 1981, International law considerations of CBD and Cartagena
ERMANZ Ethics Framework (ERMANZ 2005). Protocol.
European Union Individual nations laws (for contained experiments), EU Article 16 of National laws enforceable. Directives enforceable by EU
(EU) Directive 90/219/EC (to consult other member states that could be law. International law considerations of CBD and the
affected by an accidental release), Directive 2001/18/EC (for deliberate Cartagena Protocol.
releases, requiring submissions that allow any EU member state to do a
risk assessment). May also involve other EU decision-making processes
(e.g. for marketing of all veterinary medicinal products).
United States Food, Drug and Cosmetic Act, Plant Protection Act, Food Quality National Acts enforceable. NIH guidelines and USDA
Protection Act, National Institute of Health (NIH) guidelines, performance standards voluntary only. NEPA does not
Department of Agricultures (USDA) performance standards for fish, impose substantive requirements on any agency decision
Indian federal law, invasive species legislation, National Environmental making. NAAEC limited to effects on Canada and
Policy Act (NEPA), Endangered Species Act, North American Mexico. International law considerations of CBD
Agreement on Environmental Cooperation (NAAEC). (though not yet ratified), but not Cartagena Protocol.
582 Wildlife Research W. R. Henderson and E. C. Murphy

that less-developed countries would need substantial infra- Where to from here?
structure support in such a process. The development and release of GM biocontrol agents may be
Since the online conference, there has been no equivalent inevitable, given the increasing impacts of vertebrate pests and
forum to further discuss international consultation or biosafety the rate of progress in biotechnology. There is an urgent need for
issues. Ongoing efforts by the OIE, CBD and other agencies are researchers, policy makers and other stakeholders to consider
attempting to provide further guidance for GMO research and the international perspective and not focus solely on internal
release. For example, Norway and Canada (endorsed by the problems to look outside the national square, from social,
CBD) recently held an expert workshop on assessing risks of environmental and economic perspectives. The concept of pest
emerging GMOs (Anon. 2007). versus prized possession needs to be addressed when deciding
on whether and how to intervene in the management of animal
International issues raised by the literature populations. The recent assessment by Williams (2007) of
The current literature raises international issues of trade (where export risks of the GM contraceptive virus for house mice pro-
a country restricts imports owing to safety concerns), accessible vides one of the first examples of the types of international
pathways for transborder GMO entry (e.g. during humanitarian issues to be addressed. This assessment acknowledged the dif-
or military operations), intellectual property rights, and ethical ferent values of mice in different countries (thereby affecting the
issues. Ethical issues include who decides which species to pro- consequences of a GM virus release) and recommended further
tect and which to eradicate, rights to intervene in natural consideration of the risks. Angulo and Brcena (2007) have also
systems and rights to genetically manipulate populations to acknowledged the need for international consultation by taking
become sterile or diseased (Minteer and Collins 2005a, 2005b). an open, transparent approach to the development of the
Recent articles have highlighted the need for a consistent and Spanish rabbit vaccine.
accessible international approach for the regulation and man- Before New Zealands GM nematode for possum control
agement of GM biocontrol agents to prevent a potential politi- reaches a stage where environmental release is technically pos-
cal and/or environmental disaster (Angulo and Cooke 2002; sible, Australia should be involved in full consultation and
Anon. 2002, 2004; Nowak 2003; Gilna et al. 2005; Minteer and debate on its risks and implications (Gilna et al. 2005). The con-
Collins 2005a, 2005b; Messing and Wright 2006). They high- sultation to date has been sporadic and informal and should now
light the potential dangers of pursuing local biocontrol pro- be established on a regular and formal basis with relevant
grams without regard for conservation values in other countries. Australian agencies. Discussions should include not only
Issues of host specificity of biocontrol agents have also been environmental concerns, but also possible ramifications to
raised; they are discussed in detail elsewhere (e.g. Louda et al. trans-Tasman trade, politics, and tourism. Illegal or uninten-
2003). tional nematode transfer must also be seriously considered, as

Table 4. Summary of the online CBD conference on Biosafety considerations in the use of GMOs for management of animal populations
(CBD 2004)

Thread No. of Summary points


postings

Who and when to consult 48 Acknowledged need for an international body to consult and debate on GM biocontrol issues.
with internationally? Experts from range of disciplines (ecologists, lawyers, molecular biologists, politicians, etc.) would be useful;
possibly acting under auspices of CBD or OIE.
Need to address GM issues from perspective of different research disciplines (e.g. medicine, genetics, ecology).
Consultation should occur early in the research phase, on a case-by-case (or similar-group) basis, and with
standardised processes.
Consultation particularly important for research into disseminating GMOs.
Problems to overcome for international consensus include rights of sovereignty, differing perceptions of pest
and comfort with using GMOs per se.
Capacity to assess GMOs lacking in less developed countries.
How can we make 25 Safety mechanisms should be built into disseminating GMOs, to confine effects to target species and
disseminating target country.
GMOs safer? Existing strategies used for GM bioremediation bacteria were explained, including chemical dependence and
suicide functions.
GMOs in use of mammalian 9 GMOs potentially provide new needed control methods but there should be a clear pathway to allow
population control international consultation and evaluation.
Need policy and strong regulation in biosafety.
New challenge facing 8 High proportion of uneducated people in countries like India.
developing countries Need education, strong regulations and public participation programs.
such as India Cultural and ethical values also need to be considered.
Biocontrol 7 Potential lower-risk options exist for biocontrol designing a Trojan gene (a gene that spreads efficiently
through a population, but reduces biological fitness of the recipients), or engineering male-biased sex ratios
(daughterless technology).
Genetically modified biocontrol Wildlife Research 583

illustrated by the illegal introduction of (non-GM) calicivirus international level in the event of unauthorised GMO develop-
into New Zealand from Australia (Parkes et al. 2002). ment or release. Further social research will probably be
At a national level, jurisdiction/administration of GMO required on the public acceptability of GM biocontrols. Issues
research and release is often divided between government of resource disparities (e.g. between developed and developing
departments of environment, health and trade. There are dis- countries) will also need addressing.
crepancies in obligations to consult internationally: some coun-
tries have voluntary guidelines only, others are legally bound to Conclusions
consult with other countries. At an international level, the legal- It is clear that a single unwanted introduction of a GM biocon-
ity varies between contracting parties (at different stages of rat- trol agent could have serious consequences. Once a persisting
ification), and different countries are signatories to different transmissible GMO is released (whether intentionally, legally, or
instruments. The legal/guiding framework needs to be improved otherwise), it is unlikely that it could be completely removed
and the international procedure clarified. from the environment. The scientific community involved in
Risk-assessment processes also need to be addressed. developing GM biocontrols therefore needs to demonstrate a
Further discussions will need to include how each partys highly precautionary attitude. Scientists also have an ethical
biosafety concerns can be dealt with (e.g. engineering safety responsibility to consider the full implications of the solutions
mechanisms into GMOs and developing field-testing proto- they are researching: they must be seen to be acting openly, col-
cols), and at what point safe is considered safe enough. laboratively and responsibly. Early assessment from an interna-
Different levels of risk assessment may be appropriate at dif- tional perspective of the feasibility of releasing GM biocontrol
ferent stages of research and development (e.g. even before agents (considering technical aspects as well as possible ramifi-
proof-of-concept stage). A recent CBD-endorsed workshop on cations) may prevent valuable research funds being wasted, and
risk assessment of GM viruses (Canada Norway Expert avert environmental, economic and political repercussions.
Workshop on Risk Assessment for Emerging Applications of
Living Modified Organisms, 46 June 2007) acknowledged that Acknowledgements
there is insufficient guidance on performing risk assessments on The authors thank Brian Cooke, Robert Henzell, Chris Hardy, Lyn Hinds,
GM fish and viruses (Anon. 2007). It also acknowledged that Rod Hay, John Dowding and Greg Sherley for constructive discussions in
new methodologies may need to be developed to assist in risk this area.
assessments, because the use of field trials is likely to be prob-
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