A. WHICH ENVIRONMENTAL CONCERNS ABOUT GM CROPS ARE REALLY "SCIENCE-BASED"?
Document: REPLIES BY
Document codes: WT/DS291/R/Add.6; WT/DS292/R/Add.6; WT/DS293/R/Add.6
Complainant: Argentina
Respondent: EU
Third parties: Australia, Brazil, Canada, Chile, China, Taiwan, Colombia, El Salvador, Honduras, Mexico, New Zealand, Norway, Paraguay, Peru, Thailand, Uruguay, U.S.
Dispute stage: Mutually Agreed Solution
Agreements cited: Art. 2.2, 2.3, 5.1, 5.2, 5.5, 5.6, 7, 8, 10.1, Annex B, Annex C  Sanitary and Phytosanitary Measures (SPS)Art. III:4, X:1, X:3(a), XI:1  GATT 1994Art. 2.1, 2.2, 2.8, 2.9, 2.11, 5.1, 5.2, 5.6, 5.8, 12  Technical Barriers to Trade (TBT)Art. 4.2  AgricultureArt. I:1  GATT 1994

ANNEX H

REPLIES BY THE SCIENTIFIC EXPERTS ADVISING THE PANEL
TO QUESTIONS POSED BY THE PANEL

GENERAL COMMENTS OF THE EXPERTS

Dr. Snow
A. WHICH ENVIRONMENTAL CONCERNS ABOUT GM CROPS ARE REALLY "SCIENCE-BASED"?
1. A great deal has been written on this topic, and I will not try to summarize all of it here. Most
hypothetical concerns pertaining to environmental effects of GM crops are included in the following
list from the Ecological Society of America (Snow et al. 2005). Risk assessments are needed to
minimize the likelihood of:
• creating new or more vigorous pests and pathogens;
• exacerbating the effects of existing pests thr ough hybridization with related transgenic
organisms;
• harming non-target species, such as soil organisms, non-pest insects, birds, and other flora
and fauna;
• disrupting biotic communities, including agroecosystems;
• causing irreparable loss or changes in species diversity or genetic diversity within species.
2. Clearly, many GM crops are unlikely to cause an y of these potential problems. Furthermore,
it is not logical to group all GM crops into a si ngle category and conclude that they are either
inherently safe or inherently dangerous (see Question 103 below). It is important to evaluate new GM
crops on a case-by-case basis in each country where the crop will be grown, and to do so using
appropriate baseline comparisons. In the United States, for example, each new GM variety is
compared to its non-GM predecessor in the context of conventional agricultural practices, and in the
context of the nation's overall stra tegy for how its agricultural, environmental, and trade policies are
implemented.
3. Some of the environmental concerns that EC Me mber States have raised in the documents are
similar to concerns that routinely are addressed by regulatory agencies of the Complainants.
However, there are also major differences in re gulatory goals and policies. For example, the
conservation of farmland biodiversity is a much bigger issue in Europe than in the USA or Canada (I
am not familiar with the situation in Argentina). In some European countries, much of the non-urban
landscape consists of farmland, which is the prim ary habitat for native insects, birds, and other
animals. Also, EC Member States' strategies to manage and/or reduce herbicide applications differ
from policies of the Complainants. In addition, the EC requires post-release monitoring to check for
unanticipated environmental problems, reflecting a more cautious approach to risk assessment. The
overall philosophy of EC Directive 2001/18 differs radi cally from regulations of the Complainants by
emphasizing the possibility of cumulative and long-te rm hazards that could be caused by GM crops,
and the need for precautionary decision-making to avoid irreversible harm to the environment.
Ultimately, with regard to the Panel's questions about scientific issues, it may be useful to

acknowledge these major differences between the dis puting parties in their environmental and public
health goals and their strategies for reaching these goals.
4. Some of the delays in EC regulatory decisions c ould be related to the time needed to develop
new legislation that pertains to labeling (e.g., Regulation 1830/2003). Also, there were many requests
for new information to be added to notifications wh en EC Directive 90/220, which had been in effect
since 1990, was replaced by Directive 2001/18 in March 2001. The new Directive includes
requirements for labeling and traceability, as well as consideration of socioeconomic and ethical
issues.
5. Another question that has been raised in Europe is how to ensure that GM crops can "coexist"
with non-GM crops, including both c onventional and organic crops, such that current and future
labeling standards can be met. I will not attempt to address this socioeconomic and legal issue, other
than to note that scientific information about ge ne flow is often releva nt to questions about
coexistence. In some cases, further research may be needed to fill information gaps.
6. As a scientific advisor to the Panel, I am restricting my comments to over-arching questions
about whether various Member States had valid reasons to conclude that additional scientific research
was needed to complete their environmental risk assessments in 1998-2003.
7. Below, I list questions about possible envi ronmental problems that could result from
cultivating the specific GM crops that are cited in this dispute. These concerns apply mainly to
agricultural landscapes rather than other managed or natural habitats for flora and fauna. For each
new GM variety, one can ask whether its widesp read cultivation is likely to cause any of the
following outcomes to a greater degree than corresponding non-GM varieties:
a. create or exacerbate weed problems
b. cause direct or indirect harm to non-target species, including:
i. beneficial insects and soil organisms that affect crop yields
ii. native flora and fauna, including culturally important species
- e.g., native butterflies, skylarks, wild relatives of
crop plants
c. lead to the use of more herbicides or insecticides, including more toxic ones,
potentially harming flora, fauna, and human health.
Gene flow and the concept of "genetic pollution"

8. I will focus most of my comments on the environmental effects of gene flow from GM crops.
In this context, the dispersal of transgenes by pollen or seeds is not a problem, in and of itself, unless
this process has unwanted biological consequences. From this standpoint, the presence of transgenes
in non-GM crops, wild relatives, or weeds can be se en as being part of a normal process that occurs
with all crop genes. It is well known that crop genes can disperse widely by means of pollen and
seeds. Only certain types of new genes, such as those that confer herbicide resistance or cause harm
to non-target species, might lead to unwanted biological consequences.

9. The mere fact that GM crops are regulated may cause some people to conclude that all
transgenes are risky. I do not agree with this opinion, as I discuss in my answer to Question 103. The
process of artificially inserting genes into a plant's DNA can have unintended consequences, such as
abnormal growth or development, but it is unlik ely that 1) these effects will be ecologically
significant in commercially-produced transgenic crops, or 2) they would be more risky than the types
of side-effects that arise routinely during conventio nal breeding. In any case, many scientists agree
that genetically modified plants should be judge d on the basis of their phenotype – their actual
characteristics – rather than the pr ocess that is used to develop them (e.g., Tiedje et al. 1989, NRC
2000, Snow 2003).
10. Moreover, there is no reason to expect that tr ansgenes could be harmful simply because they
are released in a particular crop's center of divers ity (i.e., where many of the crop's wild ancestors or
original cultivars occur; e.g., Gepts and Papa 2003, Bellon and Berthaud 2004). This topic has
received a good deal of publicity, but there is no reason to expect that the effects of gene flow from
modern cultivars on the genetic diversity of wild relatives differ between transgenic vs. nontransgenic
crops. As stated above, gene flow do es not represent an environmental concern per se , unless it
results in unwanted biologi cal consequences. If there are unwanted consequences, the extent to
which the transgenes disperse via pollen and seeds may be analogous to the "exposure" term in the
simple risk assessment equation "risk = exposure x harm", in which these terms are expressed as
probabilities.
11. The documents in this dispute indicate that some Member States consider transgenes to be a
type of genetic pollution, even th ough these genes are unlikely to have any negative effects on plants,
animals, human health, or the environment. For example, Greece stated concerns about herbicide
resistance transgenes from imported oilseed rape that is not intende d for cultivation (see Question 66).
Here the perceived risk appears to be that the transgenes would be harmful to native Brassica species.
I do not agree that this would be a problem beca use 1) the imported seeds would not be cultivated
(though some mix-ups with seed sources could occur), and 2) the transgene in question – a gene that
confers resistance to the herbicide glufosinate – is unlikely to be harmful to native Brassica species in
Greece, should they eventually hybridize with crop pl ants. Even if transgen ic seeds of oilseed rape
were to leak into the local farming system, perhaps establishing volunteer populations of oilseed rape,
it is difficult to imagine how a transgene that confer s resistance to glufosinate could be harmful to
wild relatives of the crop.
B. SCIENTIFIC UNCERTAINTY DURING 1998-2003
12. Biotechnology companies have been evaluati ng characteristics of GM crops for over two
decades. As of 1998, however, the USA, Canada, and Argentina had only two years of experience
with commercial-scale cultivation of transgenic Bt and herbicide-tolera nt crops. Independent
researchers who study environmental effects of GM crops had even less experience, partly because
funding for this research was meager given the scope of key questions (e.g., Tiedje et al. 1989, Snow
and Moran-Palma 1997, Snow 2002, Snow et al. 2005). Also, it has often been impossible for
independent researchers to gain access to GM plants for field studies that precede deregulation
(Dalton 2002). Rigorous ecological research typica lly requires several years and multiple teams of
researchers to answer seemingly simple questions such as:
• Will pollen from Bt maize (corn) be harmful to native butterflies?
• Will volunteer plants from herbicide-tolera nt oilseed rape become problematic
weeds?
• How will farmland wildlife be affected by the cultivation of new GM crops as
compared to current crops?

13. Therefore, from 1998-2003, many ecological qu estions about current and planned GM crops
were just beginning to be addressed.
14. When additional scientific knowledge is needed to evaluate new GM crops, each nation's
regulators and scientific advisory committees are placed in the difficult position of choosing between
expediency and greater certainty. It is not alwa ys clear where the distinction lies between what
regulators "need to know" vs. what is merely "nice to know". Regulators often have to rely on
intuition or small-scale, unpublished studies, some of which lack adequate statistical tests (e.g.
Marvier 2002). The routine, agronomic field trials that are conducted by biotech companies prior to
deregulation are rarely intended to answer ecological -scale questions, nor would they be sufficient.
Moreover, regulators and scientific advisory committees may not have the insight to correctly identify
all of the potential risks that should be considered for each GM crop that is proposed for deregulation.
In recent years, the hotly contested political, socio-economic, and ethical questions that pertain to GM
crops have made it even more difficult to find the appropriate balance between expediency and
certainty.
15. Scientific research can play an important role in how GM crops are evaluated by confirming
or refuting widely held assumptions. Findings that are published in peer-reviewed scientific journals
help define and answer key scientific questions that relate to risk assessments. Below, I include a
sample of recent publications (1994-2003) that are relevant to this dispute because they illustrate
recent discoveries. Several of these papers challe nged widely held assumptions about pollen, gene
flow, or herbicide-tolerant crops. The context for these papers should become clear in light of my
answers to the questions from the Panel. Papers that appeared in four high-profile journals – Nature,
Science, Philosophical Transactions of the Royal Society of London, and Proceedings of the US
National Academy of Sciences – may have had the biggest impact on European decision-makers'
perceptions about environmental effects of GM crops.
Examples of recent scientific findings about pollen, gene flow, and herbicide-tolerant crops:

(Listed in chronological order; see References for complete citations.)

1. Crop genes can spread from oilseed rape to populations of weedy Brassica rapa, contrary to
previous expectations.
Jeorgensen and Andersen 1994. American Journal of Botany.
2. Transgenes that confer herb icide resistance can spread from oilseed rape to populations of
weedy Brassica rapa (not surprising based on the paper listed above, although this article
received a great deal of attention).
Mikkelsen et al. 1996. Nature.
3. Oilseed rape transgenes that confer herbicid e resistance and spread to wild relatives do not
harm weedy Brassica rapa, even in the absence of herbicide use.
 Snow et al. 1998. Molecular Ecology.
4. Transgenic pollen from Bt maize is harmfu l to monarch butterfly larvae under laboratory
conditions.
Losey et al. 1999. Nature.
In 2001, several follow-up papers in the Proceedings of the US National Academy of
Sciences (e.g., Sears et al. 2001) concluded that th is is not a problem under field conditions;

in contrast, Jesse and Obrycki (2000; Oecologia) did find evidence for harmful effects on
monarch butterfly larvae under field conditions.
5. Pollen flow among fields of herbicide-tolerant oilseed rape in Canada can lead to the
emergence of weedy volunteer plants (from unharvested seeds) that are resistant to three types
of herbicides.
 Hall et al. 2000. Weed Science.
6. A modeling study showed that more effective control of weeds, for example in fields of
herbicide-tolerant crops, could reduce the food su pply of declining populations of skylarks in
the UK.
Watkinson et al. 2000. Science.
7. Feral populations of oilseed rape can persist outside of cultivated fields for more than eight
years, contrary to previous expectations. This study showed that pers istent feral populations
can originate from earlier varieties of the crop that are no longer planted.
Pessel et al. 2001. Theoretical and Applied Genetics.
8. Transgenes were discovered in locally pro duced landraces of maize in remote areas of
Mexico, despite a national moratorium on planting transgenic maize.
Quist and Chapela. 2001. Nature.
(This finding was confirmed by further studies, while other conclusions in this controversial
paper have largely been discounted.)
9. Unharvested and spilled oilseed rape seeds can acquire secondary seed dormancy and may
persist for five years or longer in no-till as well as tilled systems, contrary to previous
expectations. Previous studies suggested th at seeds from the crop would be unlikely to
acquire secondary seed dormancy under no-till conditions.
Simard et al. 2002. Weed Technology.
10. Pollen from oilseed rape fields can travel at least three kilometers away from source fields,
thereby allowing cross-pollination to occur ove r much greater distances than previously
expected.
 Reiger et al. 2002. Science.
11. Hybridization between transgenic oilseed rape and weedy Brassica rapa is inevitable in the
UK, further demonstrating difficulties of confining gene flow if this is deemed necessary.
Wilkinson et al. 2003. Science.
12. Widespread adoption of glufos inate-tolerant oilseed rape and sugar beet might be harmful to
the biodiversity of agroecosystems in the UK, as compared to effects of currently used non-
tolerant varieties.
Results of government-sponsored Farm Scale Evaluations. Philosophical Transactions of
the Royal Society of London (several papers, including Firbank 2003, Squire et al. 2003).
13. In Canada, non-GM oilseed rape seeds that were sold to farmers were often contaminated
with small amounts of seeds that had transgenic resistance to two herbicides, glyphosate and
glufosinate. This exacerbated the emergi ng problem of volunteer canola plants with
glyphosate resistance, a trend that might jeopardize the use of low-tillage weed control.
Freisen et al. 2003. Agronomy Journal.

16. These publications illustrate that even within the specialized area of gene flow and herbicide-
tolerant crops, new findings and new unanswered questions repeatedly appeared in prominent
scientific journals between 1998 and 2003. Th is undoubtedly contributed to science-based
considerations of various Competent Authorities in Europe. Unfortunately, results from these papers
have often been distorted by the media and others, leading to a great deal of flawed information about
the actual content and the inherent strengths and li mitations of these studies. In this climate of
controversy and distrust, some of the publicity abou t risks of biotech crops has led to exaggerated
concerns about environmental risks. At the same time, the biotech industry so metimes is perceived as
being unaware of or unconcerned about valid environmental risks that are taken seriously by
professional scientists and others (e.g., Gray 2004).
17. As new publications appeared in the scientific literature during 1998- 2003, decision-makers
saw that transgenes are expected to spread very widely by means of pollen and seeds, and that
therefore these genes could show up in certified seed s sold to farmers, farmer-saved seeds, volunteer
plants, and weedy or wild relatives of the crop. Ev en in maize, which has no wild relatives or feral
populations in the USA, and which must be purchased anew each year to achieve high yields,
experience with Starlink corn showed how difficult it is to prevent unwanted genes from re-appearing
year after year. After Starlink was taken off the market, its unauthorized Bt gene (cry9C) continued to
be present in trace amounts in the US grain supply three years later (USDA 2003, Marvier and Van
Acker 2005). Evidence that transgenes could be pr esent in freely reproducing plants over very large
areas shows that "calling them back" will not be pr actical for crops like oilseed rape. Therefore,
scientific advisors of some Member States cautioned against approving certain notifications until the
environmental and economic effects of commercialization were understood more thoroughly.
18. While new questions were bei ng raised about specific GM crops during 1998-2003, many
reports stated that the GM crops grown in North America were neutral or beneficial from an
environmental standpoint. Regulatory agencies a nd biotech companies in the USA assume that
transgenic crops that are approved for commercia l production are no more ri sky than nontransgenic
crops. The US National Research Council, which is independent of the government, published two
critiques of regulatory approvals of the first GM crop varieties in the US (NRC 2000, 2002). These
critiques included suggestions for how to improve the regulatory process to make it more rigorous,
but they did not question the safety of products that had been approved for marketing. To date, no
clear environmental problems have been identified from the cultivation of GM soybean, cotton, or
maize in the USA. In Canada, one problem that has arisen from growing herbicide-resistant oilseed
rape (canola) is the emergence herbicide-resistant volunteers that are more difficult to manage than
normal volunteers (e.g., Hall et al. 2000, Friesen et al. 2003).
19. Here, it is important to recall that the pe riod during which GM crops have been grown
commercially is relatively short, having begun in 1996-1997. This is not much time for noticing
major environmental effects of GM varieties. In the USA, for instance, little attention has been paid
to documenting positive or negative effects on biodiversity empirically, and many types of effects are
not expected to be obvious to casual observers (e.g., NRC 2002, Snow et al. 2005).
C. SPECIAL CONCERNS ABOUT THE LONG -TERM EFFICACY OF BT CROPS AND THE HERBICIDE
GLYPHOSATE
20. The most widely used transgenes in commercial GM crops are various Bt transgenes and
those that confer resistance to glyphosate. These transgenes deserve special consideration because of
the environmental and health benefits they can sometimes provide, relative to current pesticide
practices, and because their effectivenes s could be lost if the target p ests evolve resistance to the Bt
pesticide or the herbicide glyphosate. This is di fferent from other types of possible risks, such as

greater allergenicity, because it deals with the loss of a benefit rather than the creation of a risk. First,
I briefly summarize the benefits of Bt crops.
21. Plant-produced Bt proteins can be more effec tive at controlling insect pests than chemical
sprays because they kill insects th at feed inside the plant, wh ere sprays cannot reach, and these
proteins are produced continuously throughout the season. In crops like cotton, which requires heavy
insecticide applications, the use of Bt cotton substitutes for many of these broader-spectrum
insecticides. This can be beneficial to farm work ers' health, populations of non-target and beneficial
insects, and the environment. Similar benefits can be seen with Bt maize, but only in regions where
insecticide use is deemed necessary to begin with. The high specificity, low toxicity, and agronomic
benefits of Bt proteins make them extremely valu able, especially in comparison to standard practices
in chemical-intensive conventional agriculture.
22. One of the risks associated with Bt crops is that target insects could become resistant to these
Bt proteins, and therefore a valuable method of insect control would be lost. Because of these
concerns, many countries in which Bt crops ar e grown have developed formal or informal
management strategies to delay the evolution of target insect pests that are resistant to Bt proteins.
23. A somewhat similar situation exists with the herbicide glyphosate (sold as Round-Up TM).
This herbicide is extremely effective at killing most species of weeds and, compared to many other
herbicides, it breaks down relatively quickly after use. Thus, it is much easier and safer to use than
herbicides such as 2,4-D, although some crops can thrive with little or no herbicide treatment
(baseline levels of herbicide use vary among crops, countries, and farmers). Glyphosate is relatively
inexpensive and it is widely available in many c ountries. In North America, large-scale conventional
farmers often use glyphosate for general weed c ontrol and for no-till and low-till cultivation, which
has several environmental benefits (e.g., reducing erosion and siltation).
24. Despite heavy use of this herbicide over the past twenty years, few weed species have
evolved resistance to glyphosate, in contrast to the large numbers of weed species that are resistant to
other herbicides like imidazolinone (WSSA 2005). The problem of allowing more and more types of
weeds to become resistant to co mmonly used herbicides is widely recognized by weed scientists,
biotech companies, and farmers, but few countries ha ve implemented effective strategies to delay the
evolution of resistance.
25. Like Bt, glyphosate has been very popular with farmers and its use has increased substantially
with the adoption of glyphosate-tolerant crops. As with Bt, one of the risks associated with the spread
of resistance genes in pests (in this case, weeds an d volunteer plants) is the loss of a benefit that is
associated with this particular herbicide. To summarize, Bt crops and glyphosate offer potential
benefits to both farmers and the environment, and scientists are skeptical about whether they could be
replaced in kind if target pests – be they insects or weeds – evolve resistance. Therefore, many
scientists, farmers, and decision-makers are work ing on methods to prolong the efficacy of Bt,
glyphosate, and glyphosate-tolerant GM crops.
1

1 References Dr. Snow:

Note: Papers cited in this report are underlined.

This reference list includes a large proportion of peer-reviewed papers about gene flow from oilseed
rape, which is the focus of my answers. Numerous publications are relevant to these questions, including
citations listed in a recent review by Devos et al. (2005).

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Dr. Squire
Notes on ecological and environmental standards
26. These notes are intended to summarise my views on some general issues pertaining to the
questions set by the panel. The notes touch on –the general weakness of current criteria and standards,
particular in relation to ecological topics, by which products of the type under review might be
judged, the importance of context and scale when as sessing new global products for specific markets,
some recent and current research in geneflow and ecological impacts.
Criteria and standards
Information required to prove that a biotech product is safe
27. No product can be assessed as being absolutely safe. There seems to be general agreement on
that. However, there is no general agreement on what should be considered as acceptably safe. As
information is first gathered on a product, the likelihood that the product is safe or not may increase
quite steeply, but gradually each further piece of information comes to contributes less and less to the
likelihood that the product is saf e. (The knowledge/safety curve assumes the form of 'diminishing
returns'). Among the present arguments, however, there seems no agreement as to when there is

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Ecology and the Environment, in press).
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enough information to satisfy all reasonable parties that a product is safe. This is largely because there
is no benchmark or set of standards by which ecological safety can be judged.
28. There is an important set of exceptions to the 'diminishing returns' shape of a
knowledge/safety curve. If the body of knowledge rev eals a new property of the biotech product, or if
the external 'environment' – the context – change s, then the knowledge/safety curve could well
steepen again because much additional knowledge is th en required to ascertain that the product is safe
in relation to the new property or context. In some instances, new properties might emerge only after a
product is grown over a period of tim e or over a wide area of land. Several of the arguments and
uncertainties in the dispute over geneflow and cont amination are of this type: current knowledge
seems insufficient to model and predict the extent of a potential problem.
International Standards and Guidelines
29. The standards and guidelines referred to in the panel's questions are (to this reviewer)
important steps towards an agreed set of standards. They differ in their stringency and required detail.
Recommendations are relatively well defined and stringent in the WTO Sanitory and Phytosanitory
Measures, in ISPM 11, and in the Guideline for the Conduct of Food Safety Assessment of foods
derived from recombinant-DNA plants . The latter in particular requires great detail in characterising
and describing the genetic material, plant metabolites and other constituents. Many of the 'requests for
further information' by the EC or its member states are consistent with (i.e. not beyond) the level of
detail indicated in that publication.
30. Less detailed and prescriptive are the 'Proposed draft annex on the assessment of possible
allergenicity.....' and the Annex III of the Cartag ena protocol on biosafety to the Convention on
Biological Diversity'. Both offer less well defined guidelines, largely because the issues (allergenicity,
impact on ecosystems) are complex and, as indicated at paragraphs 2 and 7, there are no or few
obvious quantitative criteria on which to base assessm ents. This weakness of criteria makes risk
assessment very difficult in relation to these topics.
31. As many others have said, non-biotech crop plants are not subjected to anything like the same
degree of testing as biotech plants. If they had been, the ecological standards might now be in place.
Criteria and comparators for ecological effects
32. Many of the arguments around the ecological benef its or harm of biotech crops are carried on
without reference to objective criteria. Annex III of the Cartagena Protocol on Biosafety to the
Convention of Biological Diversity indicates only the general type of information that might be
considered. Even when discussing specific products in relation to specific ecosystems, neither the
biotech companies nor the responding EC bodies refer to objective criteria when stating that a product
is safe or not. For instance, an assessing body might require more information on long-term ecological
safety without stating what type of information is necessary to ascertain safety.
33. The problems referred to in paragraph 7 arise because we lack a set of properties –physical,
chemical, biological, social and economic – which de fine that a production system is resilient and
'healthy'. As a stop-gap, ecological studies have compared GM products with non-GM isogenic lines
when making initial comparisons on a small scale, and then with another agronomic practice when
making comparisons on a field scale. Even when a comparator is used (e.g. current practice), it is not
yet clear that the comparator is an appropriate st andard, one that will sustai n crop production. Neither
side in the argument over any product has indicated, even in general terms, what a suitable set of
ecological standards might be.

Scale and context in assessing biodiversity and ecological impact
34. The arguments in paragraphs 7 and 8 are complicated further because ecological criteria will
be influenced by the context – the combination of weather, soil, landscape, the agronomy and
economics – of which the flora and fauna are an essential part. The compan ies seem to have been
unaware of, or not appreciated, the context in wh ich biodiversity exists when presenting many of the
cases in support of their products. Biodiversity is important primarily because it keeps an ecosystem
intact and functioning and makes it resilient to ex ternal perturbations. The diversity-function relation
– i.e. that diversity in microorganisms, plants a nd animals is required to maintain essential functions
such as decomposition, nutrient transformations, soil fo rmation, detoxification, regulating pests, etc. –
has been largely ignored in arguments over the safety of biotech crops (and non-biotech crops).
35. It is necessary therefore that the biodiversity in the cropped field is taken into account as
much as the biodiversity of the semi-natural habitats surrounding agriculture. Moreover
• what is considered the valuable biodiversity in one part of the world might be very
different from what is considered valuable in another part, and

• a product that is considered to have negligib le impact on total biodiversity in one part
of the world might have a large effect in another (and vice versa).

36. There are some large differences between Eur ope and North America in these respects. In
many parts of Europe, valuable biodiversity is cons idered not just that of natural and semi-natural
areas (mountains, extensive forests), but must include the flora and fauna within arable fields, both for
the reasons in paragraph 9 above (i.e. the divers ity-function relation) and because biodiversity in the
form of farmland plants, insects and birds is perceived by people as being valuable.
37. Emergent properties at the landscape scale . Moreover, it is scientifically unreasonable to
assume that an effect (positive or negative), or the absence of effect, of a biotech product when
measured at the scale of the single plant or small fi eld plot will be the same when measured at the
scale of the field, farm or landscape. Up-scaling of biological effect to the landscape was a major
topic of research in the 1990s and still is. Questions include -
• Will cross-pollination frequencies at a fiel d or feral population rise substantially as
the proportional area of a donor type increases in the landscape?

• Will volunteer and feral populations become more competitive and longer persisting
as they evolve through selection and the incorporation of new traits?

• To what extent will variation in local ph ysical conditions and occasional human error
seriously reduce the efficacy of mitigation protocols?

38. One of the main difficulties in predicting the effect of any type of innovation is still the
uncertainty over up-scaling. What will happen as more and more of the land area is affected by the
innovation? Again, progress is limited by the absence of hard criteria that define resilient or
sustainable states at the scales in question.
New research since 1998 on relevant ecological topics
39. In part to address issues in paragraphs 8 to 12, a large body of research has been
commissioned on ecological impacts and geneflow in Europe. Much of the previous research on GM

plants in the early and mid-1990s compared genetic lines with and without a GM trait, either in
containment, or in small areas of field. Howeve r, it was increasingly accepted during the 1990s that
this was insufficient to provide knowledge of the spread, persistence and ecological effects of biotech
crops at scales of the field and landscape. An im portant watershed was the symposium held in 1999 –
Geneflow in agriculture: relevance for transgenic crops – which brought together researchers from
Europe and North America to compare latest studies on geneflow and persistence of GM and non-GM
crops in the environment. This symposium drew atte ntion to the fact that geneflow at the landscape-
scale had spatial and temporal features that were not evident at smaller scales of study.
Geneflow in Agriculture: relevance for tr ansgenic crops. (1999) Proceedings of a
Symposium held at the University of Keele, 12-14 April 1999. Chaired by PJW
Lutman. British Crop Protection Council, Farnham, UK.
40. Both the EC and several EC countries have commissioned multi-partner studies on geneflow
and GM impacts. Results are reported for some stud ies; others are in progress or partly completed.
They include the following projects.
• SIGMEA – Sustainable Introduction of GM crops into European Agriculture
(http://sigmea.dyndns.org/) a EC-funded pr oject which began in 2004 and has the
main aims of (a) collating European information on the spread, persistence and
ecological effect of the main crop species under review here and (b) developing
models and decision support tools for the introduction of GM crops and their
coexistence with other forms of cropping. SIGMEA has contributions from 45 partner
organisations, and will be conducted over three years. Progress to date indicates that a
very large part of the scientific knowledge on this topic is not yet in the public
domain, because it is either recently comp leted and not published or is still in
progress.

• ECOGEN – Soil ecological and economic evaluation of genetically modified crops

http://ecogen.dk), an EC-funded project in progress, examining the effect of Bt
maize and other biotech crops on soil communities and processes at field sites in
Denmark and France.

• The Farm Scale Evaluations of GMHT crops compared GMHT and conventional
crop management on arable plants and invertebrates at more than 250 sites in the UK.
The first findings for spring-sown crops we re published in 2003 and the effects of
winter oilseed rape on biodiversity will be in 2004. Studies on gene movement and
persistence of the GMHT trait are in progress.

 The Farm Scale Evaluations of spring -sown genetically modified crops. 2003.
Philosophical Transactions of the Royal Society, Biological Sciences, 358, Theme
Issue.

41. Botanical and rotational implications of geneti cally modified herbicide tolerance in winter
oilseed rape and sugar beet (BRIGHT project). Several years of study at 5 sites in the UK comparing
the management of various herbicide tolerant varieties.
Final report at www.hgca.com and look under Crop research.
42. GMO Guidelines Project, under the auspices of th e International Organisation for Biological
Control (IOBC), funded by the Swiss Development C ooperation; the aim is to develop international

biosafety testing guidelines for transgenic pl ants, with emphasis on ecological communities and
processes in developing countries, but in principle applicable to Europe.
Hillbeck A. and Andow DA (Eds) 2004. Environmental risk assessment of
genetically modified organisms. Volume 1. A case study of Bt maize in Kenya. Eds.
CAB International, Wallingford, UK. Further case studies are in progress.
43. Additionally, there are many other country-specific studies, notably in Germany, France,
Denmark and the UK that are still in progress.
44. In summary, the understanding of environmental and ecological risk and benefit has lagged
behind the understanding of most other forms of risk . There is no set of agreed criteria for defining
'ideal' arable ecosystems and this has restricted logical and rational argument on ecological impacts in
most of the cases under review. Research is in pr ogress in a range of European contexts to provide
the scientific information which could be used to define the necessary criteria.
Annex to Notes. Persistence and movement of genes among crops, weeds and wild relatives
45. The summary in Table 1 is offered as background to several of the panel's questions that refer
to persistence and spread of GM plants. Several of the crops under re view have the potential to shed
seed and leave descendents. The following definitions are used. Volunteer(s): plants that originate
from seed or vegetative material shed or left by a crop and that inhabit fields, usually emerging as a
weed within a crop. Feral: plants th at originate from seed or vegetative material shed or left by a crop
and that exist outside fields, in waysides and the margins of agriculture. (Some authors use the term
feral for plants descended from a crop whether they are found inside or outside fields.) The table
shows that transmission among crops, feral/volunteer plants and wild relatives is highly species-
specific. Persistence and spread are generally greatest for oilseed rape, low for beet providing the crop
is prevented from flowering, very low for maize ex cept between flowering maize crops and negligible
for cotton as presently grown in EC countries.
46. A relevant factor in all arguments over biotech oilseed rape is that the perception of oilseed
rape as a weed pest in Europe has increased as more findings from research became available from the
mid-1990s and through the early 2000s. Notable facts include –
• It persists over 5 to 10 years at relatively high density in the soil (e.g. around 100
seeds per square metre).
• It is very widespread in the arable seedbank, commonly among the top 10 species
within fields and occurs frequently as a wayside plant.
• Its re-seeds in subsequent crops of oilseed rape.
• Pollen-transfer causing low-frequency cr oss-pollination occurs over several
kilometres: insect vectors are important in moving pollen.
47. For the sake of balance, oilseed rape should also be seen as a valuable and flexible 'break
crop' in cereal rotations; it needs lower agrochemic al inputs than many other crops and supports a
high level of in-field biodiversity.

Table 1. The ability of the crops to disperse genetic material through feral or
volunteer progeny, pollen movement and out crossing to form hybrids in Europe.
Author's summary.
 Beet Maize Oilseed rape Cotton

Feral / volunteer weeds
 persistence in field
 persistence outside field

medium
medium

zero1
low

high
medium

zero
zero
Pollen transfer
 crop to crop

low2

high

high

low
 crop to feral /volunteer low2 zero high zero
 crop to wild relative low2 zero low zero
Crossing frequency (on
arrival of pollen at flowering
plants)

 crop to crop high high high low
 crop to feral high zero1 high -
 crop to wild relative high (local) zero low -

 1 In most of Europe; potential for some survival in the south.
 2 Provided few or no crop plants allowed to flower.
48. The presence of a GM construct should not markedly affect the qualities in Table 1 nor the
status of oilseed rape and other crops as a weed. However, if the GM construct is accompanied by
genes that make the plant more or less male-fertile than other sexually compatible plants, then the GM
trait might be spread differentially. Oilseed rape can again be used as an example since it varies
commonly in male fertility (i.e. the proportion of pl ants that produce pollen): in some crop varieties
all (or nearly all) the plants are male fertile, but other varieties plants can be mixtures of male-fertile
(20%) and male sterile (80%) individuals. Also, volunteers (including GM volunteers) produced from
seed shed by some types of GMHT crop plant can be variously male fertile or male sterile. Generally,
crops and plants that have more male sterility get more cross-pollination from other fields or plants
than those that are fully fertile. It is importa nt to know when assessing the risk of spread and
persistence whether the variety and its offspring are fully, partly or not male fertile.
Dr. Snape
49. The comments below relate to difficulties expe rienced by the Expert in provided a full
exposition in answering questions No. 12, 16, 18 and 25, concerning the molecular characterisation of
three exhibits, EC-062, EC-063 and EC-067.
50. All the information on the molecular characterisation of a GMP (genetically modified plant)
would greatly benefit from a summarisation and compilation into a single document for each exhibit.
Currently, the evaluation of an exhibit requir es the mining of information in more than 100
attachments often representing 1000 pages. Moreover, each attachment contains a mixture of
information relative to different aspects of the risk assessment (transgene structure, expression,
phenotypic stability, toxicity or allergenicity of the product etc. …). This greatly complicates the
assessment and presents a significant risk that not a ll relevant information will be examined by each
expert. This reviewer would recommend that all information relative to an exhibit is organised

according to the structure of the EFSA-Q-2003- 005 guidance document Section III "Information
required in applications for GM plants and/or deri ved food and feed". Data initially provided by the
applicant as well as the subsequent questions and answ ers should be kept, by topic, in the appropriate
part A, B, C or D of section III. This should allow experts in different fields to have a simplified and
exhaustive access to the relevant information.
51. This reviewer has some concerns about the criteria set by in the guidance document EFSA-Q-
2003-005 section III-D2 "information on the sequences actua lly inserted or deleted". The use of the
term "insert" can be, and often has been in the e xhibits, a source of confusion. In some exhibits
"insert" is understood and presented as the DNA sequence to be introduced, in others as the integrated
transgenic coding sequence, in others as the integrated expression unit (promoter:coding
sequence:polyA signal) and in others as the entire transg enic locus. In the past, this has led applicants
to produce Southern blots and PCR-based analyses of variable quality and information content. This
has also led to a large number of subsequent request s for additional molecular information by experts.
This situation could be avoided if the information was initially requested in term of structural (and
functional) analysis of each
transgenic locus. A transgenic locus is here defined as a genetic location
where transgenic sequences are linked (i.e. co-segrega ting at meiosis when integrated into the nucleus
or remaining physically linked when part of other genetic structures). A transgenic locus can include
some plant intervening DNA between transgenic sequences. Molecular analysis in terms of a
transgenic locus is particularly rele vant for GM plants produced using Agrobacterium-based
transformation technologies as they often contain transgenic sequences dispersed over several loci
into the plant nuclear genome. Southern or PCR analyses of multi-locus GM plants can be informative
in terms of total copy numbers but have little relevance regarding the structural organisation of each
transgenic locus (copy number, rearrangement level etc…). This Expert is of the opinion that the
information required in section III-D2 should be re-organised as follows:
 (1) Sub-cellular location of transgenic seque nces (nucleus, chloroplasts, mitochondria or
maintained in a non-integrated form) [ same as part of section IIID2d in EFSA-Q-
2003-005]
 (2) When transgenic sequences are inte grated into the nucleus, the number of
independent transgenic loci. This is pr imarily obtained from inheritance pattern
determined using the structural analysis of transgenic sequence(s) in progenies
following appropriate self- or cross- pollination [ similar to part of section IIID2d in
EFSA-Q-2003-005]
 (3) Information about each transgenic locus. When more than one transgenic locus is
present in the GM plant, the molecular ch aracterisation of each locus should be
conducted independently when possible (i.e. when the GM plant is fertile).

(a) The size and copy number of all detectable transgenic sequences, both
complete and partial [same as section IIID2a in EFSA-Q-2003-005]

(b) The organisation of the transgenic seque nces at each transgenic locus and the
methods used for characterisation [ same as section IIID2b in EFSA-Q-2003-
005]

(c) In the case of deletion(s), size and f unction of the deleted regions(s) [same as
section IIID2c in EFSA-Q-2003-005]
 "Applicants should demonstrate ….. wh ich molecular analysis is but one"
[same as section IIID2 in EFSA-Q-2003-005]

(d) All sequence information including the location of primers used for detection
[same as part of section IIID2e in EFSA-Q-2003-005]
 "When events have been combined by interbreeding ….. transformation event
for risk assessment purposes" [same as section IIID2 in EFSA-Q-2003-005]
52. Please note that inheritance patterns based on the molecular analysis of transgenic sequences
in progenies are rarely presented in the exhibits. Generally, the segregation of transgene phenotype
(based on transgene expression in the progenies) is u sed to estimate inheritance pattern. However it is
well documented in the literature that the segregati on of transgene phenotype is not always a reliable
indicator of either transgene inheritance or transgene linkage as only expressing (i.e non-silenced)
transgene copies are detected by expression studies. Th is is particularly relevant when monitoring the
inheritance of non-functional transgenic sequences or when more than one copy of the transgene(s)
has been introduced into the plant genome. When a single transgene copy is integrated into the plant
nucleus, segregation studies based on transgene expr ession are satisfactory to estimate inheritance
patterns.
53. Please also note that the determination of genom ic flanking sequences is not enough in itself
to ensure that no additional unwanted DNA sequences (such as antibiotic resistance genes from vector
backbone) are present at a given transgenic locus or overall in the genome.
54. This Expert has some concerns about the cr iteria set by in the guidance document EFSA-Q-
2003-005 in sections III-D5 "Genetic stability of the insert and phenotypic stability of the GM plant".
The current requirements are to "demonstrate the inheritance pattern and stability of the trait(s)
introduced". This does not provide avenues to monitor potential recombination or deletion within a
transgenic locus across generations. This point reiterates, as expressed in 2 a bove, the need to focus
the molecular analysis at the tran sgenic locus level. It is documented that large, rearranged and
complex transgenic loci such as those generated by direct transfer of DNA (particle gun bombardment
etc…) can be prone to such genetic rearrangement s across generations. A comparative Southern blot
analysis of primary and some transgenic progeny plants using a selection of restriction digests and
probes should be adequate to assess the structural stability of each transgenic locus.
55. Overall this Expert feels that the molecular ch aracterisation of the GM plants is probably one
of the few areas where applicants have the oppor tunity to provide non-debatable and definitive
information. This information can also be obtained using routine molecular techniques at a reasonable
cost. In this context it is surprising that the opportu nity of providing a full molecular characterisation
of GM plants is not always taken by the applican ts. In the case of a small transgenic locus (up to
10,000 nucleotides) this Expert would even suggest that applicants produce a full sequence of the
transgenic locus. Most of the information n ecessary to produce an effective sequence is already
required by EFSA-Q-2003-005 (genom ic flanking sequences, architecture of the transgenic locus
determined by Southern blot analysis). This se quence of the transgenic locus and flanking plant
genomic region along with a basic inheritance study at the structural level (using dot blot or PCR) and
a Southern blot analysis of the primary tran sgenic plant and a few progenies should provide a
compelling case for the molecular characterisation of each exhibit. When a single copy of each
transgene is present in the GM plant at one locus, this analysis can be simplified by using the
segregation of transgene phenotype to estimate the i nheritance and stability of the transgenic locus.
This reviewer understands that large and more co mplex transgenic loci, such as those produced by
particle gun bombardment, may not be easy to sequence. In this later case, particular attention should
be given to the molecular stability of the transg enic locus structure over generation (see comment 3
above).

56. In conclusion I suggest:
 1. The requests from the EC for informati on from the companies could have been more
precise in its requirements, points 2 and 3 above.
 2. The information collected by the EC co uld have been compiled more systematically,
point 1 above.
 3. Companies could have provided a full mol ecular characterization of the GM plants in
the initial submissions, point 4 above.

QUESTIONS TO EXPERTS
General Questions

Question 1: On the basis of the information be fore the Panel, is there any scientific evidence to
support the hypothesis that antibiotic resist ance marker genes (ARMG) pass from a biotech
food or feed product to bacteria or other micro-organisms present in the human or animal gut?
(a) If scientific evidence indicates that such an event could occur, what risks, if any,
would arise from that event? What is the comparative relevance or magnitude
of this risk in relation to the likelihood of such a transfer from other sources of
antibiotic resistance not involving the use of recombinant DNA technology?
(b) If such risks have been identified, what is the likelihood of adverse effects to
human or animal health, in light of the processing of raw biotech products into
human food or animal feedstuffs?
(c) Are these consequences relevant to the specific types of ARMG currently used in
the products at issue in this dispute? Please explain.
(d) If such risks have been identified, what risk management options are available to
mitigate those risks and what is their efficacy?
Dr. Nutti
57. According to FAO/WHO (2000), DNA transfer fro m plants to microbial or mammal cells,
under normal circumstances of dietary exposure, would require all the following events to occur:
(a) the relevant gene(s) would have to be released, probably as a linear fragment;
(b) the gene(s) would have to survive nucleases in the plant and in the gastrointestinal
tract;
(c) the gene(s) would have to compete for uptake with dietary DNA;
(d) the recipient bacteria or mammalian cells would have to be competent for
transformation and the gene(s) would have to survive their restriction enzymes and
(e) the gene(s) would have to be inserted in the host DNA by rare repair or recombinant
events.

58. There have been numerous experiments aimed at evaluating the possibility of transfer of plant
DNA to microbes and mammalian cells. To date, ther e are no reports that marker genes in plant DNA
transfer to these cells. Even so, the use of alternative transformation methods, which do not use
ARMG, is encouraged. If alternative marker genes ar e used, they also must be evaluated regarding
their safety.
59. The transfer of marker genes which confer resistance to kanamycin, ampicillin and
streptomycin to bacteria in the human gut is unlikely to present a significant health impact since
bacteria resistant to these antibioti cs are already spread all over or are naturally found in the human
gastrointestinal tract (S malla et al., 1993; Calva et al., 1996; Shaw et al., 1993; Smalla et al., 1997).
Besides, kanamycin/neomycin and streptomycin ar e rarely used for humans due to their collateral
effects (WHO 1993).2
Question 2: On the basis of the information be fore the Panel, is there any scientific evidence to
support the hypothesis that antibiotic resistance develops through ways other than the uptake of
food or feed by humans or animals, that is, due to the potential persistence of plant-derived
DNA in the environment during crop cultivation and harvesting, and in soil residues?

(a) If scientific evidence indicates that such an event could occur, what risks, if any,
would arise from that event? What is the comparative relevance or magnitude
of this risk in relation to the likelihood of such a transfer from other sources of
antibiotic resistance not involving the use of recombinant DNA technology?
(b) Are these consequences relevant to the specific types of ARMG currently used in
the products at issue in this dispute?
(c) If such risks have been identified, what risk management options are available to
mitigate those risks and what is their efficacy?
Question 3: On the basis of the information be fore the Panel, is there any scientific evidence to
support the hypothesis that wide-spread cultivation of Bt crops such as biotech maize of the Bt
variety adversely affects non-target organisms which may be exposed to such crops under
typical agricultural practice? (See, inter alia, EC-149, EC-150, EC-151, EC-152) If so, how does
this risk compare with risks to non-target organ isms arising from non-biotech applications for

2 References: FAO/WHO. Safety asp ects of genetically modified foods of plant origin. Report of a
Joint FAO/WHO Expert Consultation on Food Derived from Biotechnology, Geneva, Switzerland, 29 May-2
June 2000. Rome: Food and Agriculture Organization of the United Nations. 2000.
www.fao.org/es/esn/gm/biotec-e.htm
CALVA, J.J.; SIFUENTES-OSBORNIO, J.; CERON, C. Antimicrobial resistance in fecal flora:
longitudinal community-based surveillance of children from urban Mexico. Antimicrob. Agents Chemother.,
vol. 40, pp. 1699-1701, 1996.
SHAW, K.J., RATHER, P.N., HARE, R.S. AND MILLER, G.H. Molecular genetics of
aminoglycoside resistances genes and familial relationships of the aminoglycosidemodifying enzymes.
Microbiol. Rev., vol. 57, pp. 138-163, 1993.
SMALLA, K.; VAN OVERBEEK, L.S.; PUKALL, R.; VAN ELSAS, J.D. Prevalence of nptII and Tn5
in kanamycin resistant bacteria from different environments. FEMS Microbiol. Ecol., vol. 13, pp. 47-58, 1993.
SMALLA, K.; WELLINGTON, E.; VAN ELSAS, J.D. Natural background of bacterial antibiotic
resistance genes in the environment. Nordic Seminar on Antibiotic Resistance Mark er Genes and Transgenic
Plants, p 8-16. June 12-13, 1997, Oslo, Norway. The Norwegian Biotechnology Advisory Board.
Workshop. World Health Organisation, Geneva.

Bt toxins (i.e., the use of Bt toxin as an insecticide in conventional and organic farming)? What
risk management options are available to mitigate any resulting risks and what is their efficacy?
Dr. Andow
Scientific evidence:
60. In the answer to this question, I will concentr ate on Bt maize and Bt cotton. Yes. there is
some scientific evidence to support the hypothesis th at wide-spread cultivation of Bt crops adversely
affects non-target organisms which may be exposed to such crops under typical agricultural practice.
However, this evidence is insufficient to establish the hypothesis that such adverse effects are
expected to occur.
61. The review of non-target effects of Bt plants in EC -149 covered 13 laboratory studies
evaluating potential hazard, and 14 field studies ai med at evaluating potential risk. Of the 13 lab
studies reviewed, I conclude that five studies did not expose the test organisms properly and therefore
are irrelevant hazard evaluations (an organism must be exposed to evaluate hazard). In two additional
studies, four of eight test species were not proper ly exposed. Of the rema ining, seven species were
studied.3 Of these seven, six were on Bt maize and none were on Bt cotton. Bt toxin had significant
adverse effects on two of these six species, the collembolan Folsomia candida (a soil organism) and
the lacewing Chrysoperla carnea. In neither case would adverse effects on these species been
predicted based on the known spectrum of toxicity of the Cry toxin. There have been no studies to
follow up the result with F. candida. Hence, Dr. Andow concludes that there is a possible hazard
(adverse effect) to collembola from Cry1Ab Bt maize, but until this is confirmed, he cannot conclude
that there is a potential risk to collembola. Ther e have been numerous studies that confirm that C.
carnea is somehow adversely affected by Cry1Ab toxin. 4 Hence, he concludes that there is a
potential risk to C. carnea from Cry1Ab Bt maize.
62. Of the 14 field studies reviewed in EC -149, six used plot sizes larger than 0.1 ha. 5 Because
most of the study organisms are mobile at spatia l scales much larger than the plot sizes used, it is
necessary to have larger plot sizes and to tailor the sampling methods to detect possible transient
differences in population sizes between treatments. Otherwise, false negatives become problematic.
The observed lack of significant differences between Bt and non-Bt treatments in nearly all the studies
with small plot sizes is difficult to interpret, b ecause the absence of an observed difference in these
kinds of experiments is not a good indication that there were no differences. The only differences

3 Folsomia candida , Coleomegilla maculata , honey bee adults, Hippodamia convergens adults, 2
parasitoid species, Crysoperla carnea.
4 Hilbeck, A., Baumgartner, M., Fried, P.M. & Bigler, F. (1998a). Effects of transgenic Bacillus
thuringiensis corn-fed prey on mortality and development time of immature Chrysoperla carnea (Neuroptera:
Chrysopidae). Environ. Entomol ., 27, 480-487. Hilbeck, A., Moar, W.J., Pusztai-Carey, M., Filippini, A. &
Bigler, F. (1998b). Toxicity of Bacillus thuringiensis Cry1Ab toxin to the predator Chrysoperla carnea.
Environ. Entomol., 27, 1255-1263. Hilbeck, A., Moar, W. J., Pusztai-Carey, M., Filippini, A. & Bigler, F.
(1999). Prey-mediated effects of Cry1Ab toxin and protoxin and Cry2A protoxin on the predator Chrysoperla
carnea. Entomol. Exp. Appl., 91, 305-316. Dutton, A. Klein, H., Romeis, J. & Bigler, F. (2002). Uptake o Bt-
toxin by herbivores feeding on transgenic maize and consequences for the predator Chrysoperla carnea. Ecol.
Entomol. 27, 441-447. Dutton, A. Klein, H., Romeis, J. & Bigler, F. (2003) Prey-mediated effects of Bacillus
thuringiensis spray on the predator Chrysoperla carnea. Biol. Contr. 26, 209-215.
5 Lozzia and Rigamonti 1998a. ATTI Giornate Fitopatologiche, 223-228. Lozzia et al. 1998. Notiziario
sulla Protezione delle plante 8: 27-39. Lozzia and Rigamonti 1998b. IOBC Bulletin 21: 171-180. Lozzia 1999.
Boll. Zool. Agr. Bachic. Ser. II 31: 37-58. Orr and Landi s. 1997. J. Econ. Entomol. 90: 905-909. Hardee and
Bryan 1997. J. Econ. Entomol. 90: 663-668.

detected in these small plot studies were in the potato experiment, 6 which will not be discussed here.
Of the remaining six, two studies had insufficient replication. 7 This low level of replication might
allow the researchers to detect only differences larg er than an order of magnitude. This detection
threshold is much larger than what would be consider ed an adverse effect, so it is possible that some
adverse effects were not detected. The remaining f our studies had an equivalent of eight replications
in each of two years. No adverse effects were observed. Unfortunately, the sampling methods were
not sufficiently tailored to detect differences in population size. Taken as a whole, excepting the
results on Bt potato, the other 13 studies would indicate that very large, order of magnitude adverse
effects on non-target species are unlikely. However, the data do not address the likelihood of other
large adverse effects on non-target organisms. Additional research would be needed before claims
about safety can be supported with scientific evidence. In particular, the cited field studies do not
enable an evaluation of the likelihood of risk to C. carnea.
63. EC-150 is a scientific paper that evaluated the effects of Bt maize litter (Bt11) on the
earthworm Lumbricus terrestris in laboratory and semi-field experiments. 8 There were no effects on
survival or growth of immature earthworms in the semi-field experiment. There were no difference in
adult survival or growth during the first 160 days in the laboratory experiment. Adults were about
20% smaller with Bt litter than non-Bt litter at 200 da ys. In total and compared with other published
results, the results of this study indicate that adverse e ffects on L. terrestris, if they exist, are likely to
be subtle. In addition, they suggest that there is a possible hazard to earthworms from Cry1Ab Bt
maize litter, but until this is confirmed, the Expert cannot conclude that there is a potential risk to
earthworms.
64. EC-151 documents a novel route of exposure of soil organisms to Cry1Ab toxin from Bt
maize.9 Cry1Ab toxin is exuded from living maize roots. There has been some controversy as to
whether the exudates are from damaged root cells or from a process involving living root cells,
however, the evidence suggests that th ere is a process involving living root cells. This result was not
anticipated. Theoretically, the Cry1Ab protein w as considered too large to be exuded from living
plant roots. This study does not document a possible adverse effect of Bt maize , however, it
demonstrates that species inhabiting the maize root rhizosphere can be exposed to Cry toxin. This is
significant because these species had been considered not at risk prev iously, and opens the possibility
that unanticipated advserse effects to rhizoshere species might be identified.
65. EC-152 demonstrates that Cry toxins persist, ac cumulate and remain insecticidal in soil by
binding to humic acids in soils. 10 Previous work had demonstrated similar results in binding to clay
in soils.11 Moreover, the toxins maintain toxicity for at least 234 days (the longest time examined). 12
Together these studies demonstrate that Cry toxins w ill persist in soils much longer than previously
believed, and that the mechanism of persistence is related to adsorption to clay particles and humic
acids in the soil. Together with EC-151, these studies demonstrate that soil organisms are likely to be
exposed to Cry toxin via root exudates and litter. None of these studies document a possible

6 Riddick et al. 1998. Annals Entomol. Soc. Amer. 91: 647-653.
7 Orr and Landis. 1997. J. Econ. Entomol. 90: 905-909 had 3 replications and conducted the experiment
in only one year. Hardee and Bryan 1997. J. Econ. Entomol. 90: 663-668 had 2 replications in each of two
years.
8 Zwahlen et al. 2003. Molecular Ecology 12: 1077-1086.
9 Saxena et al. 1999. Nature 402: 480.
10 Crecchio and Stotzky 1998. Soil Biol. Biochem. 30: 463-470.
11 Venkateswerlu and Stotzky 1992. Current Microbiology 25: 1-9. Tapp et al. 1994. Soil Biol.
Biochem. 26: 663-679. Tapp and Stotzky 1995a. Appl. Environ. Microbiol. 61: 602-609. Tapp and Stotzky
1995b. Appl. Environ. Microbiol. 61: 1786-1790.
12 Tapp and Stotzky 1998. Soil Biol. Biochem. 30: 471-476.

adverse effect of Bt maize . Together, however, they suggest th at more species in the soil may be at
risk than previously expected. These results suggest that additional studies may be needed to evaluate
these possibilities.
66. During 1999, concerns were expressed that m onarch butterflies would be adversely affected
by Bt maize pollen falling on thei r host plant, common milkweed. 13 This result was followed up with
a series of studies to assess the risk to monarchs. 14 The authors of these follow-up studies concluded
that the risk to monarch populations was insigni ficant for Mon810 and Bt11, because only a small
proportion of butterflies would be exposed to these and the toxicity of pollen was low. 15 The events
and findings associated with these studies ar e summarized in an excellent review. 16 Recent studies, 17
however, have revealed a higher toxicity of Bt po llen and anthers than found in previous studies.
Although some suggested that the risk to monarchs remains insignificant, 18 a close reanalysis of the
issues may allow other interpretations of risk to monarchs.
67. Most these same papers also addressed the pot ential risk to monarchs from Event 176 BT
maize. Although they concluded that there was no risk to monarchs from Event 176, 19 this was based
largely on the assumption that Event 176 would not be used very much in production. Had Event 176
gained greater market share, it would likely have put monarchs at risk.20

13 Losey et al. 1999. Nature 399: 214.
14 Hellmich, R.L., Sigfried, B.D., Sears, M.K., Stan ley-Horn, D.E., Mattila, H.R., Spencer, T., Bidine,
K.D., Daniels, M.J. & Lewis, L.C. (2001). Monarch larvae sensitivity to Bacillus thuringiensis -purified
proteins and pollen. PNAS , 98, 11925-11930. Oberhauser, K.S., Pr ypsby, M.D., Mattila, H.R., Stanley-Horn,
D.E., Sears, M.K., Dively, G., Olson, E., Pleasants, J.M., Lam, W.-K.F. & Hellmich, R.L. (2001). Temporal
and spatial overlap between monarch larvae and pollen. PNAS, 98, 11913-11918. Pleasants, J.M., Hellmich,
R.L., Dively, G.P., Sears, M.K., Stanley-Horn, D.E., Mattila, H.R., Foster, J.E., Clark, P. & Jones, G.D. (2001).
Corn pollen deposition on milkweeds in and near cornfiel ds. PNAS, 98, 11919-11924. Sears, M.K., Hellmich,
R.L., Stanley-Horn, D.E., Oberhauser , K.S., Pleasants, J.M., Mattila, H.R., Siegried B.D. & Dively, G.P.
(2001). Impact of Bt corn pollen on monarch butterfly populations: a risk assessment. PNAS, 98, 11937-11942.
Stanley-Horn, D.E., Dively, G.P., He llmich, R.L., Mattila, H.R., Sears, M. K., Rose, R., Jesse, L.C.H., Losey,
J.E., Obrycki, J.J. & Lewis, L. (2001). Assessing the impact of Cry1Ab-expressing corn pollen on monarch
larvae in field studies. PNAS, 98, 11931-11936.
15 Sears, M.K., Hellmich, R.L., Stanley-Horn, D.E. , Oberhauser, K.S., Pleasants, J.M., Mattila, H.R.,
Siegried B.D. & Dively, G.P. (2001). Impact of Bt corn pollen on monarch butterfly populations: a risk
assessment. PNAS, 98, 11937-11942.
16 Oberhauser, K.S. & Rivers, E.R.L. (2003). Monarch butterfly ( Danaus plexippus) larvae and Bt
maize pollen: a review of ecological risk assessment for a non-target species. AgBiotechNet, 5, 1-7.
17 Anderson, P.L., Hellmich, R.L., Sears, M.K., Sumerford, D.V. & Lewis, L.C. (2004). Effects of
Cry1Ab-expressing corn anthers on monarch butterfly larvae. Environ. Entomol., 33, 1109-1115. Dively, G.P.,
Rose, R., Sears, M.K., Hellmich, R.L., Stanley-Horn, D.E., Calvin, D.D., Russo, J.M. & Anderson, P.L. (2004).
Effects on monarch butterfly larvae (Lepidoptera: Danaid ae) after continuous exposu re to Cry1Ab-expressing
corn during anthesis. Environ. Entomol., 33, 1116-1125. Jesse, L.C.H. & Obrycki, J.J. (2004). Survival of
experimental cohorts of monarch larvae following exposure to transgenic Bt corn pollen and anthers. In: The
monarch butterfly. biology and conservation (eds. Oberhauser, K.S., Solensky, M.J.). Cornell University Press,
Ithaca, pp. 69-75.
18 Dively, G.P., Rose, R., Sears, M.K., Hellmich, R. L., Stanley-Horn, D.E., Calvin, D.D., Russo, J.M.
& Anderson, P.L. (2004). Effects on monarch butterfly larvae (Lepidoptera: Danaidae) after continuous
exposure to Cry1Ab-expressing corn during anthesis. Environ. Entomol., 33, 1116-1125.
19 Sears, M.K., Hellmich, R.L., Stanley-Horn, D.E. , Oberhauser, K.S., Pleasants, J.M., Mattila, H.R.,
Siegried B.D. & Dively, G.P. (2001). Impact of Bt corn pollen on monarch butterfly populations: a risk
assessment. PNAS, 98, 11937-11942.
20 NRC (National Research Council). (2002). Environmental effects of transgenic plants: the scope
and adequacy of regulation. National Academies Press, Washington, DC, USA.

68. As mentioned in paragraph 61, C. carnea is under a potential risk from Bt maize. Although
this has not yet been studied in the field, Dr. Andow expects that the actual risk to C. carnea would
not be large. In maize in temperate regions, this species feeds primarily on aphids, mites, thrips, and
lepidopteran eggs and larvae. Previous studies ha ve shown that the main aphid on maize does not
contain any Cry toxin when feeding on Bt maize, and that mites contain the Cry toxin, but they do not
adversely affect C. carnea. It is not known if thri ps or Lepidopteran eggs contain Cry toxin. Adverse
effects on C. carnea have been documented only when it feeds on diets containing Cry toxin or on
Lepidopteran larvae that had fed on Bt maize. Any adverse effect from Lepidopteran larvae and
possibly thrips and eggs would be diluted by feedi ng on aphids and mites. Consequently, he believes
that any adverse effect on C. carnea in the field will be subtle and difficult to detect in the field.
69. Relatively few non-target studies have been conducted on Bt cotton. This limits discussion of
potential risks.
70. It has been argued that the experience in using Bt maize and Bt cotton in the US provides
evidence that there are no adverse effects on non-target organisms. It is true that Bt maize and Bt
cotton have been used widely in the US, and it is also true that no adverse effects to non-target species
have been reported and confirmed. However, these two facts do not imply that there have been no
adverse effects on non-target organisms, because th ere is virtually no monitoring for such adverse
effects. If such effects are not looked for, it is unlik ely that they will be found, even if they exist. It
can be reliably argued that the US experience w ith these crops suggests that there have been no
immediate catastrophic adverse effects to non-target species. Such catastrophes would have become
noticeable even if they were not actively looked for. However, it is more likely that adverse effects, if
they exist, will be more subtle, and would not be readily noticed. Hence, the US experience does not
imply that there are no adverse effects on non-target species.
71. In any event, Cry toxins are un likely to adversely affect mammals through direct toxicity.
They exert their toxic effect by binding first w ith specific glycolipids in the gut. These protein-
glycolipid complexes then bind to specific receptors (cadherin-like receptors) on the wall of the gut.
The entire family of the specific glycolipid and receptor molecules are absent from mammals, thus
Cry toxins cannot act in their normal way in mammals.
72. Conclusion. Possible hazards have been identified and potential risks are evident, however, it
cannot be concluded that an actual non-target risk of Bt maize or Bt cotton has been fully
characterized. However, it is likely that if the anal ysis is completed, that there will be a documented
risk of some Bt maize events to monarch butterflies.
Comparison with Bt insecticides:
73. There are significant differences in exposure proba bility and exposure routes. Bt insecticides
have low likelihood of a non-target risk because they do not persist very long after application. The
Cry toxin is degraded rapidly under UV light, and effi cacy of the insecticide is lost usually within a
few days. This contrasts with Bt crops, which produce Cry toxin constitutively over the entire
growing season at high concentrations throughout the plant tissue, and as noted above, can persist in
the soil.
74. It is theoretically possible that hazards will be different, but this depends on the protein
structure of the Cry toxin in the plant compared with the insecticide. For example, Event 176 Cry1Ab
is significantly smaller than Cry1Ab toxin in the bacterium, Bacillus thuringiensis. The full protein in
the bacterium must be shortened to a form that show s active toxicity. Event 176 protein is already in
an active form. Thus, if the shortening process a ffects hazards, then Event 176 protein may exhibit a

different spectrum of hazards than the bacterium protein. However, it has not been scientifically
demonstrated that this difference in structure actually affects hazard. There are also differences
between Mon810 and Bt11 Cry1Ab and the Cry1Ab in the bacterium. There appears to be little
difference between Event 1507 Cry1F and the Cry1F in the bacterium.
75. Both are unlikely to adversely affect ma mmals. The Bt insecticides have undergone
significant number of studies on mammalian toxicity, a nd none have been found to be toxic to any of
the mammals studied. Coupled with paragraph 71, it is unlikely that either Bt crops or Bt insecticides
will adversely effect mammals through direct toxicity.
Risk management options:
76. Risk management measures should be commensurate to the risk.21
77. One risk management option is to prohibit the use of the Bt crop. If this option is considered,
it should be necessary to provide a worst-case risk ass essment to provide a basis for concluding that a
prohibition is commensurate to the risk. For example, what would be the harms if all earthworms
were 20% smaller after feeding on Bt maize litter for more than 160 days (see paragraph 63)?
78. Another approach would be to limit the area of the transgenic crop until a reasonable
assessment of the possible hazards and potential risks are completed. As the information is gathered,
the risk management measure can be modified consistent with the principle of "modification."22
79. For actual risks, such as the presumed risk to m onarchs, geographic restrictions in use of the
Bt crop could be considered. Incentives to use alternatives to the Bt crop can be provided.
Disincentives for the use of the Bt crop could be imposed. Monitoring of the population at risk could
be required, although this would likely be more expensive than restricting or managing use, and may
be difficult to justify based on the principle of equivalence.23
80. Efforts to improve risk assessment procedures could be viewed as a form of risk management.
Improved procedures24 would be more effective at identifying potential risk, reducing uncertainty and
allowing risk assessment to proceed more rapidly.
81. Another option would be to have no risk management. Here it seems necessary to have a risk
assessment that concludes that there are no significan t non-target risks and that unanticipated effects
will be either absent or insignificant.
82. By no means should the Panel assume that this is an exhaustive list of possible management
options. These are suggestions of approaches to risk management emphasizing avoiding risk.
Approaches that emphasize mitigation of risk or tolerance of risk could also be considered.
Dr. Squire
83. Bt crops are grown widely in some regions of the world. They are used to target crop-specific
pests, with a consequent, intended benefit that less chemical insecticide is used. They have a potential
disadvantage, in that they might harm other organisms in the ecosystem. Among these are the

21 ISPM 11, 3.4.
22 ISPM 11, 3.61.
23 ISPM 11, 3.4.
24 For example, Andow, D. A. & Hilbeck, A. (2004). Science-based risk assessment for non-target
effects of transgenic crops. BioScience, 54, 637-649.

predators and parasitoids that feed on the pests, and organisms not normally reached by chemical
insecticides. Important examples of the latter are soil fauna that might be exposed to the toxin if it
occurs in or moves out of roots, and detritus feeders and other decomposers that encounter the toxin in
dead or dying plant matter. There are therefore arguable advantages and disadvantages in the use of
the product. The emerging results from North Ameri ca show that pesticides have been reduced, but
not in all instances – context is important. Much less information on non-target organisms has come
from existing Bt maize-growing areas.
84. Potential for ecosystem effects: Assessments of potential ecological harm or benefit in areas
such as Europe have so far relied largely on findings from research in the laboratory or in contained
systems in which the Bt plant and organisms tested have no, or restricted, contact with the outside
world. Three of the exhibits referred to in Q.3 (EC-150, -151 and -152) are among this body of
research. The potential for ecological effects is indicated by these and other studies, and some of them
have shown that the toxin might well affect soil or ganisms. Exhibit EC-149 is a review of laboratory
and field data up to 1999, and concludes that, be cause of the small area and short duration of the
experiments, only limited inferences can be made a bout large scale or long term ecological effects.
Since then, there have been few field-scale experime nts on Bt crops in Europe comparable to those in
the present ECOGEN project, from which results should be available in 2005.
85. Scaling, timescales and dilution. The publishe d studies do not demonstrate one way or the
other what would happen if Bt maize were grown widely in Europe. There is no general consensus
among scientists of what would happen when a Bt crop is introduced into any new region. It is argued
by some that there could be a range of unforeseen, negative effects. In support of this view, the
authors of Exhibit EC-149 write:
Although such regulatory trials provide valuab le initial toxicity information, basing
ecological assessments solely on bi-trophic feeding trials that provide the insecticidal
protein in a highly processed form directly to the non-target organism is not
sufficient. Ecologically important interactions between plants and herbivores and
natural enemies/non-target organisms …. may be missed.
86. The implication here is that exposure to the Bt plant over wider spatial zones, and season after
season, might cause negative ecological effects that are not observed in short-term feeding trials in the
laboratory or small-plot trials in the field.
87. Conversely, it is argued that 'feeding' trials might create such artificial conditions, removing
choice of food, etc. that they greatly overestimate effects that would occur in an ecosystem. An
example of such a 'dilution' of effect is that of the monarch butterfly in North America, for which
damaging effects of Bt maize pollen, found in the laboratory, were subsequently shown unlikely to
occur in the ecosystem. There were several reasons for this: much of the insect's food plant occurred
outside Bt maize fields, the seasonal timing of the insect and the release of pollen only partly
overlapped and the actual exposure of the insect to th e toxin was lower in maize fields than in the
laboratory. A lay article Butterflies and Bt corn : allowing science to guide decisions, published by
several US universities and the USDA provides a summary of the study; scientific papers are
assembled in the journal Proc. Natl. Acad. Sci. USA, vol. 98, 2001. The results of many other feeding
experiments would similarly be 'diluted' (this review er believes) when examined in the context of the
ecosystem.
88. The general absence of ecological standards and comparators. As discussed in the
introductory notes, and in relation to herbicide-tolera nce (Q.6), a context and comparator are vital to
any assessment, and as in the case of herbicide-tolerance, they have rarely been taken fully into

account when assessing Bt crops. Assessments purporting to show harm, for instance, should compare
the effects of the Bt crop with an existing practice or standard, such as current pesticide management,
or options using application of Bt preparations (for which there appears little ecological information).
As for assessment of GMHT crops, there is no definiti on or set of criteria for an acceptable, resilient
and sustainable ecosystem. It should be feasible for scientists to define such criteria, which would be
generic but with location-specific elements. A recent case study of Bt maize in Kenya shows what can
be done quickly by combining global and location-specific knowledge (Hilbeck A & Andow DA, eds.
2004. Environmental risk assessment of genetically modified organisms. Vol 1, A case study of Bt
maize in Kenya. CABI Publishing).
89. Mitigation. It much more easy to confine maize within cropped fields than it is oilseed rape or
beet, for example, so the ecological risks are mainly to do with the in-field food web and with any
wide ranging organisms that visit the field (i.e. th ere are no risks in much of Europe through ferals
and volunteers). To plan a mitigation strategy need s understanding of the existing state of the food
web. In the UK Farm Scale Evaluations (see Notes) , which used >60 fields of GMHT, not Bt, maize,
the maize fields supported a small set of organisms compared to the organisms in oilseed rape and
beet. This was so because the persistent herbicid es then used on maize sev erely reduced the arable
plants available for the insects to eat. Moreover, only a few species in the above-ground food web
were found on the maize itself. If maize is adopted over wide areas of a country where it had not been
grown, that itself would affect biodiversity whether or not it was Bt maize. Any additional effect of Bt
should be seen in relation to the effect cause d by the introduction of maize as a crop species.
However, if Bt maize was found to harm the in-field biodiversity, it should be feasible to reverse the
effect by growing different crops in a rotation. Perh aps the main uncertainty is the effect of Bt maize
on the soil food web. As indicated, the ECOGEN project should inform on this. Effects in the soil
would be harder to monitor because it is very di fficult to assess soil biodiversity, particularly among
bacteria and fungi. Nevertheless, any adverse effects should be mitigatable through a rotation.
90. Q.3 addresses a complex topic. The following is offered in summary.
• The evidence is insufficient to confirm whether widespread cultivation of Bt crops in
Europe would affect non-target organisms.
• The weight of evidence from contained expe riments and small-plot field experiments
generally favours the view that little immediate toxic effect of Bt maize on the food
web will occur in the field; but the eviden ce admits the possibility of chronic effects
following long-term exposure to Bt maize.
• The question could only be answered by long-term study of the effects of Bt maize
and a comparator on the complex interacti ons among non-target organisms; it would
be difficult to see how that would be done without intensely monitored, field-scale
experimental plantings in Europe.
• Criteria for resilient food webs would need to be proposed and agreed as a standard
for any new research: without such criteria, progress is impossible.
Question 4: On the basis of the information be fore the Panel, is there any scientific evidence to
support the hypothesis that the wide-spread cultivation of Bt maize or other, non-biotech
applications of Bt toxins, leads to the emergen ce of Bt-resistant target organisms under field
conditions? If so, what risk management options exist to mitigate any resulting risks and what is
their efficacy?

Dr. Andow
91. Evidence for resistance. Widespread non-biotech applications of Bt toxins have resulted in
the Bt-resistant target pest organisms under field conditions. 25 Probably the best studied example is
diamondback moth on cabbage, which has developed fiel d resistance to Bt insecticide applications in
several countries, including the US (Hawaii), Japan, Philippines and elsewhere.
92. There is strong evidence that resistance will de velop in the field to any insecticide applied
uniformly over wide areas for a long enough period of time. 26 This has been a scientific consensus
since the 1980s.
93. Bt-resistant target organisms have been f ound – pink bollworm (Pectinophora gossypiella) to
Bt cotton in the field,27 but this has not yet led to field failures.28
94. Evolutionary theory predicts that the evol ution of resistance will proceed as directional
selection, at least up to the point resistance becomes common. This means that resistance in the field
cannot be prevented, it can only be delayed.
95. There is a scientific consensus that resistance to Bt maize is inevitable, 29 even though it still
has not been detected in the field.30 This is based on the preceding empirical and theoretical evidence.
96. Generally speaking, of all of the potential environmental risks of transgenic Bt crops, it can be
said that resistance in the target pests is a real, ta ngible risk, while risks associated with gene flow and
risks to non-target organisms are mostly only potential risks.
97. There are several risk management options for Bt maize, but they are all concerned with
reducing the selective advantage of resistant alleles in populations of the target species.31
98. The most widely used resistance management strategy for Bt maize (and all Bt crops) has
been the high-dose/ refuge strategy. 32 This has been used by th e US Environmental Protection

25 Tabashnik, B.E. (1994). Evolution of resistance to Bacillus thuringiensis. Ann. Rev. Entomol., 39,
47-79.
26 NRC (National Research Council). (1986). Pesticide resistance: strategies and tactics for
management. National Academy Press, Washington, DC.
27 Tabashnik, B.E., Patin, A.L., Dennehy, T.J., Liu, Y.B., Carrière, Y., Sims, M.A. & Antilla, L.
(2000). Frequency of resistance to Bacillus thuringiensis in field populations of pink bollworm. PNAS, 97,
12980-12984.
28 Tabashnik, B.E., Carrière, Y., Dennehy, T.J., Morin, S., Sisterson, M.S., Roush, R.T., Shelton, A.M.
& Zhao J.-Z. (2003). Insect resistan ce to transgenic Bt crops: Lesson from the laboratory and field. J. Econ.
Entomol., 96, 1031-1038.
29 Tabashnik, B.E. (1994). Evolution of resistance to Bacillus thuringiensis. Ann. Rev. Entomol., 39,
47-79. Gould, F. (1998). Sustainability of transgen ic insecticidal cultivars: integrating pest genetics and
ecology. Ann. Rev. Ent., 43, 701-726. Andow, D.A. 2001. Resisting resistance to Bt corn, pp. 99-124. In
D.K. Letourneau and B.E. Burrows (eds.), Genetically engineered organisms: Assessing environmental and
human health effects. CRC Press, Boca Raton, FL.
30 Bourguet, D., Chaufaux, J., Séguin M., Buisson, C., Hinton, J.L., Stodola, T.J., Porter, P., Cronholm,
G., Buschman, L.L. & Andow, D.A. (2003). Frequency of alleles conferring resistance to Bt maize in French
and US corn belt populations of the European corn borer, Ostrinia nubilalis. Theor. Appl. Genet., 106, 1225-
1233.
31 Gould, F. (1998). Sustainability of transgenic insecticidal cultivars: integrating pest genetics and
ecology. Ann. Rev. Ent., 43, 701-726.

32 A major factor determining resistance management is the "dose" of the Bt crop. Dose is defined as a
concentration of Cry toxin in the Bt crop plant in relatio n to the expression of the resistance phenotypes. For a
high dose, the concentration of Cry toxin must be sufficie ntly high that the plant tissue is toxic enough so that
resistance is functionally recessive (Fig. B1, concentration HD or higher, Taylor & Georghiou 1979; Tabashnik
& Croft 1982). Any other concentration is a low dose. A high dose is desired for resistance management
because resistance can be delayed more readily than fo r a low dose by using the high-dose/ refuge resistance
management strategy. A refuge is an area where the pest is not subjected to selection from the Cry toxin and
can produce a viable population. The same crop plant without the Bt gene can function as a refuge as long as Bt
insecticides are not applied to it. Th ree conditions are essential to the success of the high-dose/ refuge strategy:
resistance is recessive, resistance is rare, and there is sufficient intermating between adults coming from refuges
and Bt fields.
The recessivity of resistance is determined by complex interactions (Tabashnik & Croft 1982). If
resistance is determined by allelic variation at a single locus, with R designating the resistance allele and S the
susceptible allele, dominance of the R allele is related to both the mechanism of resistance in the insect and the
toxin concentration in the plant (Fig. 1, appendix to question) . For example, if resistance in the insect is related
to a change in a receptor in the midgut lining, resistance may be physiologically recessive (Fig. 1a, appendix to
question), while in other cases, the dose-mortality curve for RS heterozygotes may be physiologically co-
dominant (Fig. 1b, appendix to question) or dominant. In both cases, the toxin concentration in the plant will
also determine whether the R allele functions as a recessive allele ( HD in Fig 1a and 1b, appendix to question).
If the concentration is not high enough relative to the dose-mortality curve for the RS heterozygote, the Bt crop
would be a low dose crop. Clearly, high dose cannot be determined if resistance ha s not been observed in the
target pest.
Recessive resistance to Bt cotton (Cry1Ac) has been found in cotton budworm ( Heliothis virescens
(F.), Gould et al. 1997, Gahan et al. 2001), cotton bollworm ( Helicoverpa armigera (Hübner), Akhurst et al.
2003) and pink bollworm ( Pectinophora gossypiella (Saunders), Tabashnik et al. 2000), but no resistance has
been found in cotton bollworm ( Helicoverpa zea (Boddie)). Resistance to Bt maize, which is the most widely
grown Bt crop, has not been found in the widely distributed European corn borer ( Ostrinia nubilalis (Hübner),
Bourguet et al. 2003) or in southwestern corn borer ( Diatraea grandiosella (Dyar)). Resistance has been found
in the beetle Chrysomela tremulae F. to Bt poplar (Genissel et al., 2003), but not in rice stemborer (Scirpophaga
incertulas (Walker), Bentur et al. 2000) to Bt rice. Neither Bt poplar nor Bt rice is planted commercially at this
time.
There have been several attempts to designate a high dose without reference to a resistance allele,
because the observed resistance alleles may not be the onl y ones and resistance has not been observed in some
species. The EPA-SAP (1998) has suggested that 25 times the the concentration of toxin at which 99% of the
individuals in a susceptible population die (LC
99, lethal concentration 99) may function as a high dose. Caprio
et al. (2000) argue that 50 times the LC 99 is a better supported by the published data on insecticide resistance.
The value of the EPA-SAP (or Caprio et al. 2000) standard is that it is readily operationalized for transgenic Bt
crops. The fundamental problem with this approach, however, is that it does not anticipate novel resistance
mechanisms and may contribute a false sense of security about the validity of the high-dose assumption.
If resistance is recessive, it challenging to measure when rare. As a rule of thumb, for the high-dose/
refuge strategy to be effective, the frequency of resi stance should be <0.001 (Roush & Miller 1986). This
implies that resistant phenotypes will be extremely rare, <1 x 10 -6, and that more than 10 6 individuals from
natural populations must be screened to estimate such low allele frequencies.
Refuges are areas of habitat within normal dispersa l distance of the target species from a Bt habitat
where the R and S alleles have similar fitness. In addition, an effective refuge must be able to support a viable
population of SS homozygotes in the tran sgenic landscape. A refuge must be within normal dispersal distances
so that there is sufficient mixing and mating between individuals emerging in the refuge and Bt field. A refuge
can be any habitat where the target species occurs, including the non- Bt crop, other crops, and other non-crop
plants, as long as the R allele does not have a selective advantage in these fields. If the refuge areas cannot
support a viable population of SS homozygotes, then the population tends toward extinction, and there are
insufficient SS individuals available to delay resistance evolution (Ives & Andow 2002).
References cited. Akhurst, R.J., James, W., Bird, L. & Beard, C. (2003). Resistance to the Cry1Ac δ-
endotoxin of Bacillus thuringiensis in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae). J.
Econ. Entomol., 96, 1290-1299. Bentur, J.S., Andow, D.A ., Cohen, M.B., Romena, A.M. & Gould, F. (2000).
Frequency of alleles conferring resistance to a Bacillus thuringiensis toxin in a Philippine population of a rice

Agency (US-EPA). For Bt maize, it is generally accepted that a 20% refuge is needed for the high-
dose/ refuge strategy to be effective.33
99. Low-dose Bt events will require a greater refuge. There is no scientific consensus as to how
big a refuge is needed. One approach, used by the US-EPA is to require a 20% refuge during the
initial period of use, when market penetration remains low. This may provide enough time for the
scientific evidence to accumulate so that an approp riate refuge size can be determined. A different
approach was used by Australia, where a very large refuge was required initially, which was gradually
reduced over time to a 70% refuge.
100. Additional resistance management strategies may become possible to develop after resistance
in the target pest is identified and characterized.
101. The efficacy of the high-dose/refuge strategy cannot be assessed empirically in the field.
Indeed, it would probably be unethical to conduc t such a field experiment because it would be
necessary to have a positive control treatment where resistance was encouraged . Resistant insects in
such a control treatment could escape, leading to field failures that undermine the efficacy of the high-
dose/ refuge strategy.

stem borer, Scirpophaga incertulas (Lepidoptera: Pyralidae). J. Econ. Entomol., 93, 1515-1521. Bourguet, D.,
Chaufaux, J., Séguin, M., Buisson, C ., Hinton, J.L., Stodola, T.J., Porter, P., Cronholm, G., Buschman, L.L. &
Andow, D.A. (2003). Frequency of alleles conferring resistance to Bt maize in French and US corn belt
populations of the European corn borer, Ostrinia nubilalis. Theor. Appl. Genet., 106, 1225-1233. Caprio, M.A.,
Sumerford, D.V. & Sims, S.R. (2000). Evaluating tran sgenic plants for suitability in pest and resistance
management programs. In: Field manual of techniques in invertebrate pathology (eds. Lacey, L.A. & Kaya,
H.K.). Kluwer Academic Publishers, Amsterdam, pp. 805-828. EPA-SAP (US Environmental Protection
Agency – Scientific Advisory Panel). (1998). Transmitta l of the final report of the FIFRA Scientific Advisory
Panel Subpanel on Bacillus thuringiensis (Bt) plant-pesticides and resistance management. Meeting held on
February 9-10, 1998. Report dated, April 28, 1998. (Docket Number: OPPTS-00231). Gahan, L. J., Gould, F.,
& Heckel, D. G. (2001). Identification of a gene associated with Bt resistance in Heliothis virescens. Science,
293, 857-860. Génissel, A., Augustin, S., Courtin, C., Pi late, G., Lorme, P. & Bourguet, D. (2003). Initial
frequency of alleles conferring resistance to Bacillus thuringiensis poplar in a field population of Chrysomela
tremulae. Proc. R. Soc. Ser. B.-Bio., 270, 791-797. Gould, F., Anderson, A., Jones, A., Sumerford, D., Heckel,
D.G., Lopez, J., Micinski, S., Leonard, R. & Laster, M. (1997). Initial frequency of alleles for resistance to
Bacillus thuringiensis toxins in field populations of Heliothis virescens. PNAS, 94, 3519-3523. Ives, A.R. &
Andow, D.A. (2002). Evolution of resistance to Bt crops: Directional selection in structured environments.
Ecol. Lett., 5, 792-801. Roush, R.T. & Miller, G.L. (1986). Considerations for design of insecticide resistance
monitoring programs. J. Econ. Entomol., 79, 293-298. Tabashnik, B.E., Patin, A.L., Dennehy, T.J., Liu, Y.B.,
Carrière, Y., Sims, M.A. & Antilla, L. (2000). Frequency of resistance to Bacillus thuringiensis in field
populations of pink bollworm. PNAS, 97, 12980-12984. Tabashnik, B.E. & Croft, B.A. (1982). Managing
pesticide resistance in crop-arthropod complexes: Interactions between biological and operational factors.
Environ. Entomol., 11, 1137-1144. Taylor, C.E. & Georghiou, G.P. (1979). Suppression of insecticide
resistance by alteration of gene dominance and migration. J. Econ. Entomol., 72, 105-109.
33 EPA /US Environmental Protection Agency). (2001) . Biopesticides registration action document –
Bacillus thuringiensis plant-incorporated protectants. 16 October 2001. [WWW document] URL
http://www.epa.gov/pesticides/biopesticides/pips/bt_brad.htm (last accessed 23 November 2004).

102. Some greenhouse experiments have confirmed the efficacy of the high-dose/refuge strategy.34
103. Theory has predicted that the high-dose/ ref uge strategy will be efficacious when resistance
allele frequency <0.001, 35 resistance is recessive, and there is sufficient adult movement between the
refuge and Bt fields to ensure matings between resistant and susceptible phenotypes.
104. These assumptions have been confirmed scientifi cally for European corn borer (Ostrinia
nubilalis) in the northern US corn belt. These assumpti ons are likely for Europ ean corn borer in the
southern US and in western Europe. They have not been confirmed for southwestern corn borer
(Diatraea grandiosella) in the southern US, for European corn borer in eastern Europe, or for
Mediterranean corn borer (Sesamia nonagrioides) in southern Europe. Hence, it can be concluded
that in some circumstances it is possible to predict that the high-dose/ refuge strategy should be
efficacious. It is possible that the absence of detec tion of field resistance to Bt maize in the US is
partially attributed to the efficacy of the high-dose/ refuge strategy.

34 Shelton, A.M., Tang, J.D., Roush, R.T, Metz, T.D. & Earle, E.D. (2000). Field tests on managing
resistance to Bt engineered plants. Nature Biotechnology, 18, 339-342.
35 Roush, R.T. & Miller, G.L. (1986 ). Considerations for design of insecticide resistance monitoring
programs. J. Econ. Entomol., 79, 293-298.

Appendix

Figure 1. High dose (HD) and low dose (LD) concen trations (dotted vertical lines) in relation to
hypothetical mortality of SS, SR, and RR insect genotypes as a function of Bt crystal protein
concentration (solid diagonal lines). a. R allele is physiologically recessive, so SS and RS mortality is
similar. b. R allele confers an intermediate, co-dominant level of physiological resistance, so RS
mortality is about midway between SS and RR mortality.

SS RS
RR 50%
90%
99%
10%
1%
HD2
3
4
5
6
7
8
-2 -1 0123
Mortality (probits)
LD
a.
2
3
4
5
6
7
8
-2 -1 0123
Concentration (log scale)
Mortality (probits) RS
HD LD LD
SS RR 50%
90%
99%
10%
1%
b.

Dr. Squire
105. The emergence of resistance by pest insects to pesticides differs widely from context to
context depending on factors such as the exposure to and strength of the toxin, the movement of insect
populations from areas where the pesticide is not applied, and the genetics and mating system of the
insect. Resistance to Bt crops has occurred and is infl uenced by the 'dose' of toxin delivered to the
pest and the genetic nature of the pest, among ot her factors. Because of long-standing scientific
interest in pest resistance generally, knowledge has been imported from that body of work to construct
models for the evolution and mitigation of Bt resi stance. The processes involved in Bt resistance and
its management are generally appreciated by scientis ts, and mitigation strategies that have a strong
scientific basis have been considered. By the la te 1990s, mathematical models of the population
dynamics of resistant and susceptible biotypes were being used to estimate the number of years for
which the GM trait would remain effective in the f ace of genetic adaptation by the pest. The point is
that sound scientific knowledge has been applied to this problem for several years.
106. A strategy for mitigation needs protocols for both pre-emptive management, testing for
resistance and managing it if found. The strategies might involve providing refuges in or near the crop
where resistant individuals do not have an advantag e over resistant ones, periodically controlling the
pest by other means and growing different 'types' of Bt maize, or Bt and non-Bt maize, either together
or in sequence. Mitigation will be much easier to implement where the existing ethos is sympathetic
to integrated pest management based on genetic and ecological principles. A useful, recent summary
of the topic, much of it in layman's terms, directed at Bt maize in Kenya but based on generic
arguments, is given by: Fitt GP et al., (2004). R esistance risks and management associated with Bt
maize in Kenya. In Hilbeck A & Andow DA, eds. 2 004. Environmental risk assessment of genetically
modified organisms. Vol 1, A case study of Bt maize in Kenya. CABI Publishing.
Question 5: On the basis of the information be fore the Panel, is there any scientific evidence to
support the hypothesis that Bt maize varieties a re any more toxic to humans or animals than
conventional maize under field conditions? If so, what risk management options exist to
mitigate any resulting risks and what is their efficacy?
Dr. Nutti
107. Based on the information before the Panel, ther e is no evidence to support the hypothesis that
Bt maize varieties are more toxic to humans or animals than conventional maize under field
conditions.
Dr. Squire
108. (This response does not address the part of this question relating to humans. Toxicity was
considered as part of the Response to Q.3 and some of the argument here is repeated.) Assessing a
product's toxicity to ecological processes is usually more complicated than assessing the toxicity of,
for example, a feedstuff for a domestic or farm an imal. The domestic or farm animal may have the
food under test as its sole or main diet in reality, whereas non-target organisms generally have a wider
choice of food throughout the habitat. Also, the ex pression of a toxin in a plant and its effect on
animals eating the plant are influenced by local gr owing conditions, the weather, the behaviour or the
local animals, etc., all of which differ between s ites. Therefore making inferences about toxicity to
animals in one part of the world (e.g. a maize fi eld in a European country) from information in a
constrained experiment, or field experience in anothe r part of the world, should be done with great
caution. The contention that nothing adverse has occurred when growing Bt maize in, say, North

America, should not be taken as definitive evidence by itself that nothing will occur in Europe, where
the organisms and their interactions are different.
109. Feeding experiments in which organisms are expos ed to live or dead Bt plant matter, and
have no other choice of food, should be viewed as providing information only on the possibility that
the Bt plant might be toxic to the animals in the fi eld. Many of the studies in contained experimental
systems are of this type. Some do and others do not indicate harm occurred, but none should by
themselves be taken to indicate that harm will nec essarily occur in the field. (Several major crop
species would fail toxicity tests of this type.) Ma ny of the plot-scale experiments examined in EC-149
were criticised by the authors of that review for a range of reasons, but taken as a whole, experiments
at the scale of small field plots and one season's exposure indicate no acute toxicity to animals other
than the pest.
110. Evidence, either way, of harm to animals of growing Bt crops in the field in Europe is
therefore inconclusive at present. The question could be approached by a range of experimental
studies over longer periods, but the results would need to be compared against the effects of other
crop species on the animals, and against independent criteria defining a normal or healthy state for the
animals in question (as for Q.3).
Question 6: On the basis of the information be fore the Panel, is there any scientific evidence to
suggest that herbicide tolerant crops (whether biotech or developed through mutagenesis) are
more persistent in the agricultural environment or more persistent in the non-agricultural
environment than their conventional counterparts? If so, do herbicide tolerant crops qualify as
a potential "pest" as the term is used in the International Plant Convention's (IPPC)
International Standard for Phytosanitary Measures (ISPM) 11 (EC-130)?
(a) What is the potential for the establish ment and spread of herbicide tolerant
plants arising from handling, spillage during transport of the plant/plant parts,
or any other means outside of cultivation in the absence of application of the
herbicide? How is any potential for establishment and spread affected by
environmental conditions, the presences of wild or conventional relatives of the
herbicide tolerant plants in an area, or other factors?
(b) What is the potential for the establish ment and spread of herbicide tolerant
plants in the presence of herbicide app lication (in fields, urban, domestic or
other environments)? How is any potential affected by the existence of feral
related plant species; infertile wild relatives; seed survival in relevant
pedoclimatic conditions; the reproduction biology of the species; or other
factors?
(c) Is this potential different for biotech crops tolerant to two wide-spectrum
herbicides? Please explain.
(d) If significant risks of establishment and spread have been identified, what risk
management options exist to mitigate any resulting risks and what is their
efficacy?
(e) What types of post-market monitoring and data collection activities could be
envisaged on the basis of the monitori ng and review principles described in
ISPM 11?

Dr. Nutti
111. According to the International Plant Conve ntion's (IPPC) International Standard for
Phytosanitary Measures (ISPM) 11 (EC-130), the definition of pest is "Any species, strain or biotype
of plant, animal or pathogenic agent injurious to plant or plant products". In my opinion, herbicide
tolerant crops can not be qualified as "pest", according to this definition.
Dr. Andow
112. I will provide a theoretical response to the ma in question and address only part (e) of the
subparts of this question. I will use the seven kingdom taxonomy here and throughout in discussing
organisms.
113. Persistence of herbicide tolerant crops (wheth er biotech or developed through mutagenesis)
will be determined by the relative fitness of the crop with the trait and the degree of release of the
trait. The only way that a GMHT would be more persistent than a non-GMHT crop would be if it had
a higher relative fitness and/or it was released at a higher rate. Both factors would have to be assessed
on a case-specific basis. For GMHT maize, cotton, soybean and beet, the question is moot because
there are no comparably good non-GMHT varieties. For GMHT oilseed rape, evidence from Canada
suggests that the GMHT varieties are used more co mmonly than the non-GMHT varieties, thus there
is some scientific evidence to suggest that GMHT oilseed rape may be more persistent in the
agricultural or non-agricultural environment than their conventional HT oilseed rape.
114. A similar argument holds for persistence of he rbicide tolerant crops (whether biotech or
developed through mutagenesis) compared to thei r conventional counterparts. When examining
release rate, however, the relevant release rate for the conventional counterpart is that of a single
variety.
115. Annex 2 of ISPM 11 specifies "Phytosanitary ri sks from LMOs may result from certain traits
introduced into the organism, such as those that in crease the potential for establishment and spread, or
from inserted gene sequences that do not alter the pest characteristics of the organism by that might
act independently of the organism or have uninten ded consequences." Thus phytosanitary effects of
the transgene outside of the original organism of introduction are covered by ISPM 11.
116. Annex 2 elaborates on this further in the next clause, which states "In cases of phytosanitary
risks related to gene flow, the LMO is acting more as a potential vector or pathway for introduction of
a genetic construct of phytosanitary concern rather than as a pest in and of itself. Therefore the term
"pest" should be understood to include the potential of an LMO to act as a vector or pathway for
introduction of a gene presenting a potential phytosanita ry risk." Phytosanitary risks associated with
gene flow and persistence are covered by ISPM 11.
117. Moreover, Annex 3 "Determining the potential for a living modified organism to be a pest",
specifies that "potential phytosanitary risks fo r LMOs may include: a. Changes in adaptive
characteristics which may increase the potential for introduction or spread, … b. Adverse effects of
gene flow or gene transfer. This indicates that th e potential for introduction and spread are a part of
phytosanitary risk, as well as adverse effects of gene flow.
118. From the perspective of gene flow and persistence, GMHT crops may qualify as a potential
pest according to ISPM 11, as long as they can be sh own to be released at a higher rate and/or have a
selective advantage compared to a conventional counterpart.

119. Of course it is possible for a GMHT crop to qualify as a potential pest according to ISPM 11
if persistence is expected to be the same as a conventional counterpart. This would occur if it
potentially resulted directly or indirectly in so me adverse effect that was different from the
conventional counterpart. For example, if contamin ation of conventional crop germplasm (related to
the "coexistence" issue) were cons idered an injury to the conventi onal plant, then there would be a
reason to believe that all GMHT crops can be c onsidered potential pests under ISPM 11. All other
potential pest risks would probably have to be considered on a case by case basis.
120. ISPM 11 is ambiguous whether all possible e ffects of GMHT crops can be considered
phytosanitary risks. The definition of "pest" is given on page 6 of ISPM 11 as "Any species, strain or
biotype of plant, animal or pathogenic agent injurious to plants or plant products [FAO, 1990; revised
FAO, 1995; IPPC, 1997]" The concept of "injurious to plants includes environmental risks, as
indicated in supplementary text S1.
121. Annex 2, which covers the scope of IPPC for LMOs states "PRA may constitute only a
portion of the overall risk analysis for import and release of a LMO. For example, countries may
require the assessment of risks to human or animal health, or to the environment, beyond that covered
by the IPPC." This is a clear acknowledgement that there are some environmental risks that are not
covered by the IPPC.
122. Annex 1 addresses the scope of the IPPC in regard to environmental risk. This text is
reproduced here:
123. "The full range of pests covered by the IPPC extends beyond pests directly affecting
cultivated plants. The coverage of the IPPC definition of plant pests includes weeds and other species
that have indirect effects on plants, and the Conven tion applies to the protection of wild flora. The
scope of the IPPC also extends to organisms which are pests because they:
– directly affect uncultivated/unmanaged plants

124. Introduction of these pests may have few commercial consequences, and therefore they have
been less likely to be evaluated, regulated and/or placed under official control. An example of this
type of pest is Dutch elm disease (Ophiostoma novo-ulmi).
– indirectly affect plants

125. In addition to pests that directly affect host pl ants, there are those, like most weeds/invasive
plants, which affect plants primarily by other pro cesses such as competition (e.g. for cultivated plants:
Canada thistle ( Cirsium arvense) [weed of agricultural crops], or for uncultivated/unmanaged plants:
Purple loosestrife (Lythrum salicaria) [competitor in natural and semi-natural habitats]).
– indirectly affect plants through effects on other organisms

126. Some pests may primarily affect other organi sms, but thereby cause deleterious effects on
plant species, or plant health in habitats or ecosystems. Examples include parasites of beneficial
organisms, such as biological control agents.
127. To protect the environment and biological di versity without creating disguised barriers to
trade, environmental risks and risks to biological diversity should be analyzed in a PRA."

128. In considering indirect effects, Annex 1 a ddresses "indirect effects on plants" and "indirect
effects on plant through effects on other organisms." Indirect effects are not defined, but the two
examples provided are two of the five fundamental two-species interactions in ecological science,
consumption (parasite-host or predator-prey or herbivore-plant), competition, mutualism, amensalism,
and commensalism. Thus, indirect effects occur via any combination or pathway of these to-species
interactions. This is the standard interpretation of the concept of indi rect interactions which give rise
to indirect effects in the field of ecology. As the IPPC is science-based, I assume that standard
scientific usage is appropriate for interpreting key terms taken from science.
129. Under this interpretation (and ot her interpretations as well), the effects of the herbicides
applied to the GMHT crops is not covered under A nnex 1 and can not be considered a phytosanitary
risk. Herbicide use cannot be considered a direct or indirect effect of the GMHT crop, because
humans apply the herbicides. It is inaccurate to say that this is an indirect effect of the GMHT crop
through effects on other organisms. The GMHT crop does not affect humans to apply herbicides.
Causality works the other way. Humans affect th e distribution of the GMHT crops and at the same
time affect herbicide use. Thus , the effects of herbicide use on GMHT crops is not covered under
Annex 1.
130. Section 2.3 of ISPM 11 covers "Assessment of potential economic consequences" and also
covers environmental risk. Under 2.3.1.2, indirect pest effects are considered. This is a list of
potential endpoints to consider given that it is an indirect effect. Thus, section 2.3 does not provide an
interpretation to cover the effects of herbicides applied to GMHT crops as a phytosanitary risk.
131. It would be disingenuous to suggest that herb icide effects should be considered as if they
were independent of the use of a GMHT crop. While it may be true that the environmental effects of
herbicides are regulated under independent regulatory structures, this does not change the fact that
they are used part and parcel with GMHT crops because that is their intended use. Thus GMHT crops
almost always will be accompanied by the use of the specific herbicide.
132. The only ecologically consistent position would be to identify humans as the potential pest.
This, however, is absurd under ISPM 11.
133. Thus I conclude that GMHT crops can qualify as a potential "pest" as the term is used in the
International Plant Convention's (IPPC) International Standard for Phytosanitary Measures (ISPM) 11
(EC-130). However, not all of the risks associated with GMHT crops can be considered phytosanitary
risks.
(e) Post-market monitoring and data collection:
134. Section 3.6.1 of ISPM 11 describes the monitoring and review of phytosanitary measures.
This concentrates on the principle of "modification", and is repeated here.
3.6.1 Monitoring and review of phytosanitary measures
The principle of "modification" states: "As conditions change, and as new facts
become available, phytosanitary measures shall be modified promptly, either by
inclusion of prohibitions, restrictions or requirements necessary for their success, or
by removal of those found to be unnecessary" (ISPM N° 1: Principles of plant
quarantine as related to international trade).

Thus, the implementation of particular phytosanitary measures should not be
considered to be permanent. After app lication, the success of the measures in
achieving their aim should be determined by monitoring during use. This is often
achieved by inspection of the commodity on arrival, noting any interceptions or any
entries of the pest to the PRA area. The information supporting the pest risk analysis
should be periodically reviewed to ensure that any new information that becomes
available does not invalidate the decision taken.
135. This suggests that an important part of th e risk management measures applied to GMHT
crops should include a process to review the necessity of the management measures. Given that many
ecological effects are scale-dependent, manifesting more readily at larger spatial and temporal scales,
some management measures could be linked to the sp atial extent of use of the GMHT crop. If the
spatial extent of use is not great enough, then certain management measures could be considered
unnecessary. Alternatively, if the GMHT trait persis ts in a given locality for long enough, additional
management measures could become necessary.
136. In a similar fashion, after certain time and area thresholds have been exceeded, it would be
reasonable to review management measures to determin e if the measures need to be dropped, retained
or strengthened. Such thresholds should be agreed upon in advance.
137. Finally, for precautionary management measur es, there should be certain time and area
thresholds specified which would trigger a review to determine if sufficient information had
accumulated to merit a reduction in the level of precaution. Presumably the precautionary
management measures would provide data from which it would be possible to assess their necessity.
Dr. Snow
138. Note: I use the terms herbicide resistance and herbicide tolerance interchangeably. Weed
scientists generally use the term resistance in this context to di stinguish between a newly evolved
condition and an existing level of tolerance that has always been present. Biotech companies prefer to
use the term tolerant for transgenic crops that have new genes for protection from herbicides.
139. ISPM 11 definition of a pest (explained further in Annexes 1-3 of ISPM No. 11):
• "Any species, strain, or biotype of plant. ... injurious to plants or plant products,"
• "... many organisms indirectly affecting plan ts also satisfy this definition (such as
weeds/invasive plants)"
• Annex 3 mentions LMOs with increased pot ential for spread du e to ... "pesticide
(including herbicide) resistance or tolerance"
140. In summary, a crop with herbicide tolerance can qualify as a potential "pest" under the
ISPM 11 definition if 1) it transmits herbicide resi stance to weedy volunteer plants or the crop's
sexually-compatible wild relatives, and 2) this leads to worse weed problems. Volunteer oilseed rape
and its sexually-compatible relative, Brassica rapa, are considered to be w eeds because they compete
with food crops and require management. These plants could become more abundant if they gain one
or more types of herbicide resistance from the crop. This can occur when the herbicide in question is
used extensively (e.g., glyphosate) and herbicide-resistant volunteers become much more difficult to
control.

141. The magnitude of the problem, if and when th ere is a problem, depends on whether resistant
volunteers and related weeds can be killed using othe r herbicides, crop rotation, and/or tilling, and on
what amount of effort is needed to keep these plants from harming crop yields (e.g., Warwick et al.
1999, Friesen et al. 2003). This problem varies among species of crops and weeds that occur in
different countries. Brassica rapa is known to h ybridize with oilseed rape in Europe, but its
abundance is usually kept under control by tilling and herbicide applications.
142. I conclude that there is no evidence to suggest that herbicide-resistant plants are more
persistent in non-agricultural areas where herbicides are not used (e.g., Crawley et al. 2001), and there
is no scientific reason to expect that this could ha ppen. Also, each type of herbicide-tolerant crop
should be considered individually – it is not possible to generalize across all herbicide-tolerant crops.
143. ANSWERS 6a. and 6b: Note: I assume that this question refers to plants that are cultivated
by farmers, rather than seeds that are imported exclusively for processing.
Routes of dispersal
144. Seeds – Genes for herbicide resistance, whether transgenes or not, can spread by means of
seed movement. Herbicide-tolerant crop plants can easily spread by seed dispersal that occurs during
handling, transport, spillage. Depending on the crop, seeds from volunteers, feral populations, and
weedy relatives of the crop also disperse widely by wind, water, farm vehicles, and the transport of
seeds, forage, and manure.
145. Pollen – Like other crop genes, transgenes for he rbicide tolerance can spread to sexually-
compatible crops and wild or weedy relatives that occur nearby by cross-pollination. Therefore, the
potential for spread and establishment of transgen es is greater when similar crops or wild/weedy
relatives occur nearby.
146. Pollen-mediated transfer of crop genes to ot her fields and to sexually-compatible wild
relatives occurs more frequently in crops that typi cally cross-pollinate, such as maize and to a lesser
extent oilseed rape, than in soybean, which is mostly self-pollinated. With respect to oilseed rape, the
weed Brassica rapa is an obligate outcrosser that is known to hybridize with the crop (e.g., Jeorgensen
and Andersen 1994).
147. Transgenic tolerances to both glyphosate (Round-Up) and glufosinate (Liberty, Basta) are
inherited as dominant, Mendelian traits. Some transgenic varieties may have more than one insertions
of a gene for herbicide tolerance, while others have just one. A plant needs just one copy of the gene
at one insertion site to be tolerant of the herbicid e in question. This means that pollen from the crop
can transfer the herbicide tolerance trait to seeds in other farmers' fields and to seeds of volunteer crop
plants, related weeds, or wild relatives. When different crop varieties have different genes for
herbicide tolerance, weed populations can acquire sev eral types of herbicide resistance genes, as
shown by Hall et al. (2001) for weedy, volunteer oilseed rape in Canada.
Potential for establishment
148. Some crops establish volunteer and feral populations more easily than others, depending
largely on how thoroughly domesticated the crop is. Oilseed rape is not highly domesticated in the
sense that it can survive as free-living, volunteer populations that can re-seed from year to year (e.g.,
Pessel et al. 2001). Also, viable seeds of oilseed rape can persist in the soil for several years, and then
germinate and reproduce (Simard et al. 2002). In contrast, maize does not establish feral populations

in Europe. At most, maize plants can be present as volunteers in the year following the crop, but they
do not establish problematic, long-term populations.
149. Hybridization with sexually-compatible weed y relatives can further allow transgenes to
spread and persist in and around agricultural fields. Even when first-generation offspring between a
crop and a weed are inferior to normal weeds, crop genes can often become incorporated into the local
gene pool of the weed following further cross- pollination. If progeny from these crosses are
completely infertile, they will not produce seeds a nd they will not be a problem unless they establish
clonal, vegetatively propagated populations, as in perennial grasses.
Further spread after establishment
150. Frequencies of specific crop genes in free-livi ng plant populations depend on their rates of
introduction and also their effects on plant fitness (i.e, relative survival and reproduction). Unlike
some types of nontransgenic herbicide tolerance, the transgenes that confer tolerance to glyphosate
and glufosinate are not expected to have any negative effects on crop yields or the fitness of crop
relatives (e.g., Snow et al. 1998).
151. In the absence of exposure to the herbicide in question, herbicide-tolerant plants will not have
any selective advantage over their non-transgenic c ounterparts. But when the herbicide is used
repeatedly, it will select very quickly for plants that are resistant to the herb icide. The scientific
literature in weed science is full of examples of rapid evolution and spread of herbicide resistant
weeds (e.g., Warwick et al. 1999, WSSA 2005).
152. ANSWER 6c: In principal, the potential for the establishment and spread of herbicide-
tolerant plants is similar for nontransgenic vs. transgenic crops that have genes for these traits. Rates
of establishment and spread of herbicide-tolera nt crops are affected by whether the genes for
herbicide resistance show dominant or recessive inheritance, whether the resistance genes are
associated with fitness costs (i.e., reduced survival or reproduction), and the extent to which the plants
are exposed to the herbicide(s) in question. Transgenes that confer resistance to glyphosate or
glufosinate show dominant inheritance and they are not likely to be associated with fitness costs (e.g.,
Snow et al. 1998).
153. In answers 6a and 6b, I discuss how biotech crops with resistance to either of these two wide-
spectrum herbicides might contribute to the spread and establishment of herbicide-resistant weeds.
This can happen by gene flow to related weeds, and also by another mechanism – spontaneous
selection for weeds that are naturally resistant to the herbicide. The second mechanism has occurred
in only a few species for the two herbicides in que stion. Once a gene for glyphosate or glufosinate
resistance is present, is likely to increase in fre quency in compatible weeds and volunteer populations
that are exposed to these herbicides. The more the herbicide is used, the stronger the selection
pressure favoring herbicide-resistant weeds. Gly phosate use is already common and is expected to
increase further when crops are bred to have transgenic resistance to this herbicide. Glyphosate is
very useful to farmers because it is wide-spectrum, killing most weed species very efficiently, and it is
considered to be "environmentally friendly", as compared to other herbicides. Glufosinate is used less
widely, but its use also is expected to increase after biotech crops with glufosinate tolerance have been
commercialized.
154. ANSWER 6d: Several risk management options are available to mitigate problems that result
from plant pest populations that are tolerant of one or more herbicides, but they may or may not be
very practical (e.g., Beckie et al. 2004, Devos et al. 2005). One way to prevent herbicide-tolerant
weeds from becoming more abundant is to rotate among herbicides with different modes of action

from one year to the next, in a cycle that accounts for the persistence of viable seeds of herbicide-
resistant weeds in the soil.
155. Another method for suppressing populations of herbicide-resistant weeds is to use several
types of herbicides in tank-mixes each year, before or after a crop is grown, to kill off resistant plants.
Management options become more challenging and more complicated when the pest population has
genes for several types of herbicide resistance. In some cases, it may be necessary to revert to the use
of herbicides that have greater toxicity and longer persistence in the environment (e.g., 2,4-D).
156. The efficacy of different management options will vary depending on local conditions and the
types of crops and rotation systems that are used. For oilseed rape, which has a long-lived seed bank,
seeds of weedy volunteer plants may emerge over a period of at least 4-5 years (Simard et al. 2002).
157. As mentioned above, it is expected that transgenes that confer herbicide tolerance will not be
associated with negative effects on the weedy plants that inherit them. This me ans that the transgenes
could persist indefinitely in weed populations, even in the absence of exposure to herbicides.
158. ANSWER 6e: In cases where the persistence and proliferation of herbicide-resistant weeds
could pose problems, post-market monitoring could be used to develop an early warning system for
alleviating the problem. Weed scientists are already acutely aware of emerging problems with
herbicide-resistant weeds that develop in response to heavy herbicide use (e.g., WSSA 2005). Many
farmers are also familiar with these problems. Stewardship programs can be developed for risk
management, for example by encouraging farmers to report cases of herbicide failure to the proper
authorities. Studies of reported cases could be carried out to test for resistance due to transgenes, and
to determine whether the stewardship programs are working as planned.
159. However, by the time the spread of herbicide resistance is detected in free-living pest
populations, such as weeds or volunteer plants, it may be too late to prevent these plants from
proliferating further. By this time, continuous e fforts may be needed to keep their populations as low
as possible. After a transgene for glyphosate has spread widely in oilseed rape, as occurred in
Canada, there is no easy way to eliminate the problem.
Dr. Squire
160. The background to this question is that several of the crops have become widespread feral
plants or weeds (see Notes, Annex). Oilseed rape has been grown in Europe for centuries, but has
become a weed in many parts of Europe following a great increase in its cropped area since the 1970s.
It is now a regular and persistent member of th e arable and wayside seedbank (buried viable seeds).
Oilseed rape is not considered a problem on waysid es and is ousted by perennial vegetation such as
grass. Within fields, it is treated as part of the broadl eaf weed flora. It also contributes to yield of the
next oilseed rape crop, but has generally not been considered a problem in this respect, since the
quality of oil has generally been similar in the volunteers and crop plants. Similarly weed beet, arising
from seeding crop beet, is common in beet-growing areas and brings similar but lesser problems.
161. Are HT plants a potential pest? Oilseed rape and beet are both crops and pests, as are many
other crop plants (pests as defined in ISPM-11, pa ge 6). The volunteers or ferals arising from HT
crops would be no greater pests than those arising from non-HT crops, unless the specific herbicide
was used to favour them as pests, or their presence in the field or in the yield itself had greater
significance than the presence of non-HT. Since agriculture began, crops and weeds of the same
species have existed side by side and exchanges ge nes – but this does not mean that volunteer/feral
weeds are not pests.

162. More serious problems to do with their persistence as pests will arise if labelling or marketing
'rules' specified that a non-biotech crop had to be of a high degree of purity, i.e. to contain less than,
say, 0.9% of GM in the yield.
163. 6a. In the absence of use of the specific herbicide, HT plants would have the same potential as
non-HT, as indicated in 6 above. Species would diffe r as in Table 1. Establishment and spread
depends on local conditions in ways that are not clear. For example, feral wayside populations of
oilseed rape differ greatly in number in areas st udied in France and the UK. Also, wild, weedy
relatives differ in their presence and number in different parts of Europe, as do hybridisation rates
between crops and between crops and wild relatives. But the essential point is that – in the absence of
the specific herbicide to which the plants are tolera nt – plants having HT and non-HT traits should act
similarly.
164. 6b. Suppose that HT plants were growing in mixed vegetation with non-HT plants of the
same and other species, then applying the herbicide would favour the HT plants, lead to greater seed
set on HT plants and their increase in the seedbank. This would happen wherever the plants were
growing and whether the HT trait was in volunteer/ feral plants of had been transferred to wild
relatives. Whether such differences occurred in practice would then depend on the how widely the
specific herbicides were used (which itself would vary between countries and crops).
165. 6c. In the absence of the specific herbicide, the potential for establishment and spread should
be similar for biotech plants tolerant to the wide-spectrum herbicides glufosinate ammonium and
glyphosate. The genes conferring tolerance to either s hould not enable the plants to be better at, say,
persisting in the soil. Relative persistence between the two GMHT types would differ if one or both of
the herbicides was used. As indicated, this will be highly specific to the agronomy of a farm or region.
For instance, in the UK, glyphosate has risen in the last few years to be the second (or maybe first)
most widely applied herbicide in arable fields, while glufosinate ammonium is used little in fields or
waysides. If GMHT glyphosate tolerant plants were present in fields, then they would be advantaged
at this specific time and area, where glufosinate ammonium tolerant plants would not.
166. 6d. Risk management options should be similar to those recommended for non-biotech
varieties (oilseed rape and beet), but would need to be more stringently applied if cropping was to
ensure the proportion of GM in non-GM remained be low a threshold. Establishment can be reduced
by –
• Reducing or preventing seed loss to the soil, e.g. by harvesting oilseed rape before too
many pods split (not always possible), uprooting flowering beet plants.
• Not cultivating soil after harvest, leaving seeds to germinate on soil and then killing
them by cultivation or herbicide spray.
• As part of normal broadleaf weed control, ensure they do not flower and seed in later
crops of other species.
• Spread can be reduced by –
• Clearing seed from combine harvesters and other machinery between crops.
• Transporting seed from field to market in sealed containers or vehicles.
• Cutting or spraying roadsides that support la rge populations of feral oilseed rape, but
these practices will impinge on the semi-natural flora.
• Using a pollen barrier to reduce field-to-field geneflow (oilseed rape and maize), such
as a strip of flowering crop 50 to 100 m wide between the donor and recipient crops
and which is not harvested as part of the recipient crop.

167. The main difficulty in preventing establishment and spread is with oilseed rape, specifically
in that it will enter the soil and become dormant ev en if soil is not cultivated after harvest, it will
flower and seed, usually unseen, in a later crop of oil seed rape and it is impractical to remove all seed
from farm machinery. Lesser problems occur with weed beet.
168. 6e. Post-market monitoring: ISPM 11 does not give great detail on monitoring. The type of
monitoring in this instance would differ depending on purpose: for example, monitoring the effect of
a HT cropping on biodiversity or ecosystem f unctioning would require a different set of
measurements from monitoring the presence or abundance of a HT trait. What is clear from existing
data and work in progress is that monitoring of this type is far from simple and needs much time and
effort, especially if the aim to measure low frequenci es (e.g. 1%, 0.1%) or small effects (e.g. 1.5-fold
effects on populations or ecological processes). Recently introduced populations are highly
aggregated or clumped and this increases the number of samples and area over which samples have to
be taken. At present, there are no reliable and accepted monitoring schemes for the presence or impact
of biotech-derived plants: research is in progress to develop such schemes (see Notes, paragraph 14),
but it is not even certain that it would be feasibl e or practicable to monitor low frequency occurrences
or ecological effects routinely in the field.
Question 7: On the basis of the information be fore the Panel, is there any scientific evidence to
support the hypothesis that repeated use of a given biotech herbicide tolerant crop has adverse
effects on flora and fauna, including soil micro - and macro-fauna? If so, how does this compare
with any similar risks of adverse effects from the repeated use of a non-biotech herbicide
tolerant crop (i.e, one developed through mutagenesis)?
Dr. Andow
169. I will restrict this answer to the following ge netically modified herbicide tolerant (GMHT)
crops: oilseed rape, fodder beet, cotton, maize, soybean, and sugar beet. I will use the seven kingdom
taxonomy in discussing organisms.
Adverse Effects of GMHT Crops:
170. There is abundant evidence that repeated use of a given biotech herbicide tolerant crop would
likely result in the evolution of resistance in weeds to the herbicide. While it can be debated if this
adverse effect results from repeated use of the GM HT crop or from the repeated use of the herbicide
of which the GMHT crop is tolerant, in practice th e two are so tightly correlated, and the GMHT crop
is expected to have the herbicide applied to it. Hence a risk assessment should consider both as
correlated causal factors. In the language before the panel, weed resistance is a risk resulting from the
associated changes in agricultural practices.
171. Adverse effects on non-target flora and fauna could arise directly from transgene products,
directly from the herbicide compounds, or indirectly through the effects of the transgenic crop or the
herbicide on the environment. Although the author s of the Farm-Scale Evaluation (FSE) Trials of
GMHT crops in the UK36 concluded that all adverse non-target effects arise indirectly from the effects

36 Brooks et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1847-1862. Champion et al. 2003, Phil. Trans.
R. Soc. Lond. B, 358, 1801-1818. Firbank 2003, Phil. Trans. R. Soc. Lond. B, 358, 1777-1778. Haughton et al.
2003, Phil. Trans. R. Soc. Lond. B, 358, 1863-1877. Ha wes et al. 2003, Phil. Trans. R. Soc. Lond. B, 1899-
1913. Heard et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1819-1832. Heard et al. 2003, Trans. R. Soc. Lond.
B, 358, 1833-1846. Roy et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1879-1898. Squire et al. 2003, Phil.
Trans. R. Soc. Lond. B, 358, 1779-1799.

of the herbicide on the environment, 37 this highly credible argument is not proved conclusively. 38 In
any event, the FSE Trials demonstrate that some, but not all, GMHT crops can have adverse effects
on non-target organisms. I believe that it is likely that additional adverse effects on non-target
organisms may be reported in the future.
172. Gene flow from a GMHT crop to a weedy relative can create weeds that are more difficult to
control with herbicides. This occurred for GMHT oilseed rape in Canada. GMHT maize, cotton and
soybean have not yet been grown legally in loca tions where there are weedy relatives and none occur
in Europe, the US, Canada or Argentina, so this risk is not possible in these countries. Beets have
wild relatives in Europe, and weedy beets have developed as a consequence of gene flow in the past.
173. Gene flow from a GMHT crop to another crop cultivar can contaminate seed supplies (part of
the "coexistence" issue) or reduce genetic diversity in the crop. Contamination is a serious concern,
although it can be debated if this adverse effect is on the flora (the crop) or the growers of the crop. It
is clear that the harm is to the growers of the crop. Genetic diversity of cotton, maize and soybean is
not known to be great in Europe, so this is unlikely to be a concern. It may be possible that there are
significant sources of genetic diversity in oilseed rape and beets in Europe.
174. To my knowledge there are no reports of adverse effects on soil micro- or macro-flora or
fauna separate from those in the UK-FSE trials. Nor are there any reports of adverse effects on soil
dwelling bacteria, algae, or protozoa. However, to my knowledge there have not been any studies of
any of these possible effects. The Panel should not infer that the absence of information implies an
absence of effect.
175. To my knowledge, all reports have not found adverse effects on flora or fauna from antibiotic
resistance genes or gene products. Extensive st udies on nptII did not find any adverse effects, and
found that any undetected adverse effects would like ly be several orders of magnitude smaller than
naturally occurring phenomena.
Comparison with non-GM HT Crops:
176. There are several ways in which GMHT and non-GMHT crops may differ that are significant
to risk to flora and fauna.
177. GMHT traits are typically hemizygous, which means that there is only one copy of the locus
in the plant. Normal plant genes occur in two copies, that are either different (heterozygous) or the
same (homozygous). Gene flow to wild relatives or other crop varieties will result in hemizygous
offspring for GMHT traits, which are likely to ex hibit dominant expression. For non-GMHT traits,
hybrid offspring are most likely to be heterozygo us and may exhibit varying degrees of dominance.
This means that whatever the selective advantage of the trait in the original HT plant, it is likely to be
similar for GMHT hybrids, but possibly less for non- GMHT hybrids. Because selective advantage is
one of key determinants of spread of genes, this would suggest that GMHT traits could be more likely
to spread than equivalent non-GMHT traits.39
178. The promoters on most GMHT transgenes keep th e transgene turned on and expressing at all
times. The non-GMHT transgenes are regulated by plant promoters, which may or may not keep the

37 Champion et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1801-1818.
38 Andow 2003, Nature Biotechnology, 21, 1453-1454.
39 It should be recognized that while the bits of information that comprise this argument can be found in
the information before the Panel, the argument itself cannot be found in the information before the Panel.

gene turned on all the time. This may enable the herbicide to be used for a longer period of time on
the GMHT crop than the non-GMHT crop. If so, gr eater degrees of weed reduction may be possible
in GMHT than non-GMHT crops, which would r esult in greater non-target effects and higher
selection for weed resistance in more species of weeds.40
179. Most of the GMHT crops tolerate glufosinat e and glyphosate, while most of the non-GMHT
crops tolerate imidazolinone and sulfonylurea herbic ides. It is possible that there are important
differences in these herbicides that result in different risks to flora and fauna. I am unaware of studies
that address this possibility. In addition, the opti ons for managing resistant weeds are likely to differ
among the different herbicides.
180. Scale of use has a significant effect on risk. 41 Some GMHT crops have gained significant
spatial scales of use that dwarf those of non-GMHT crops. For example, RoundUp Ready ® soybean
now occupies >60 of the US soybean area (>20 million hectares), while no non-GMHT trait has come
close. With greater scale, small effects can reinforce each other and become more apparent. This
would be especially true for non-target effects, such as those found in the FSE trials.
181. If contamination is an adverse effect on flor a, then GMHT crops have this risk while non-
GMHT crops do not. This is based on the definition of harm, which is premised on distinguishing
between GM and non-GM traits.
182. If all of the factors mentioned in paragraphs 177 through 181 are equivalent between the
GMHT and non-GMHT crop, then there will be no pred icted difference in the risks of adverse effects
from the repeated use of a GMHT or non-GMHT crop.
Dr. Squire
183. The difficulties of argument in this topic revolve around what is classed as an adverse effect
(Notes, paragraphs 32-35). There are no objective criteria for what is an ideal state and what therefore
might be judged an adverse effect against that stat e. At best, HT cropping has been compared against
an existing cropping system. Certainly in Europe, the important and relevant flora and fauna should
include those within the managed areas of fields.
184. The Farm Scale Evaluations (FSE) in the UK (Notes, paragraph 14) detected small, but
important shifts caused by using GMHT rather than the conventional management, and illustrate
issues of context. The difference in weed management rather than the GM or conventional crop plants
per se was the effective agent. The direction of effect differed depending on the severity of the
conventional management. In maize, where convent ional management uses pe rsistent, highly toxic
herbicides, GMHT increased the flora and fauna; in spring oilseed rape and beet, where current
practice was less effective against arable plants, it had the opposite effect, reducing the flora and
fauna. The crucial point about the FSE – and one that has been missed or misrepresented by many
international commentators – is that the effects on the arable flora or weeds (though small by
international standards) were important in the context of the UK's arable scene in the early 21 st
century. The flora and fauna of arable fields were important in the national biodiversity as perceived
by many people; these flora and fauna had already b een severely depleted by intense agriculture, so
further depletion was unnecessary and unacceptable.

40 It should be recognized that while the bits of information that comprise this argument can be found in
the information before the Panel, the argument itself cannot be found in the information before the Panel.
41 NRC (National Research Council). (1987). Introduction of recombinant DNA-engineered
organisms into the environment: key issues. National Academy Press, Washington, D.C.

185. In principle, there should be no difference between the effects of GMHT cropping and non-
biotech HT cropping, though this has not been compared on a suitable scale. The BRIGHT project
(Notes, paragraph 14) made some comparisons of GMHT and non-GM HT crops.
186. A general knowledge of the macro- and micro-fa una in the soil suggests they will be much
less sensitive to GMHT cropping than will the above-ground flora and fauna. The FSE and some other
work included macrofauna caught on the soil surface, but no large-scale studies of herbicide tolerance
have included soil micro-and macro-fauna. (ECOGE N will provide information on soil communities
at three field sites for Bt maize.) All evidence from small-scale work in field plots and containers
points to the herbicides glyphosate or glufosinate a mmonium being much less directly toxic to soil
fauna than other agrochemicals and the GMHT pl ants themselves having no effect on soil organisms
different from that of non-GM plants.
Question 8: What are the different detection methods currently available for testing for the
presence of material from genetically modified plants?
(a) Have commercially available detection methods changed since the mid-1990's?
Were methods available in the mid-1990's event specific?
(b) Please outline the steps necessary to va lidate a detection method, including the
determination of what types of reference materials needed and differences in
validation steps for qualitative and quantitative detection methods.
(c) What are the differences in the intended uses of qualitative and quantitative
detection methods? What are the diff erences between event specific and non-
event specific detection methods? How does the availability of different types of
detection methods relate to risk assessment and risk management processes?
Dr Nutti
187. Answer 8a: Beginning in the 1990's, commercial protein-based and DNA-based detection
methods have become available. Both methods can be used in a qualitative and a quantitative way.
Early DNA-based methods have been mainly desi gned as qualitative methods only. More recently,
there has been a shift towards quantitative PCR me thods. Today, most published methods are based
on real-time PCR technology that can be operated bot h in a qualitative and a quantitative way. Some
DNA-based methods were designed for the identification of a specific event while others were
designed to identify only a certain vector c onstruct or a common genetic element (e.g., 35S
promoter). Today, the development of these three different types PCR methods continues. The
selection of method depends on the intended purpose. The experience for protein-based methods is
similar to the experience with DNA-based methods – there are many commercially available protein-
based methods also. These methods can confirm the presence or absence of specific proteins
expressed by the different biotech events. These protein-based methods can be applied in both a
quantitative and qualitative fashion. The protein based methods, while not event-specific, can identify
uniquely most commercial events.
188. Answer 8b: I would like to inform that I am not an expert in this area, and I will list bellow
the information that I could find in order to help the panel, but is important to point out that I will list
the references and information founded and not express my personal opinion.

Interlaboratory studies:
189. The International Union of Pure and Applied Chemistry (IUPAC) classify interlaboratory
studies into the following three categories:
 1. Method performance: Determines the bias and precision of an analytical method.

2. Material performance: Assigns a valu e and an uncertainty (or reliability) to a
characteristic (usually concentration) of a material.
 3. Laboratory performance: Permits the ev aluation of each participant against preset
criteria or criteria estimated from the study itself.

190. Although the procedures regarding statistical data evaluation from these three types of
interlaboratory studies may be identical, the use and interpretation of the resulting statistical estimates
will be determined by the primary purpose of the study.
Validation of methods:
191. The validation of methods consists of two phases. The first is an in-house validation of all of
the parameters except reproducibility. The second is a collaborative trial, the main outcome of which
is a measure of the repeatability and reproducibility in order to estimate the transferability of methods
between laboratories. Usually a small-scale colla borative trial should be performed to test the
ruggedness of a particular method before the expense of organizing a large-scale trial is incurred. In
case any improvement of the method or the method description is needed, only limited costs are
incurred through the pre-trial, while a failure of a full interlaboratory method validation due to an
ambiguous method description is a very costly failure. Implementation of an already validated method
in a laboratory needs to include the confirmation that the implemented method performs as well under
local conditions as it did in the interlaboratory me thod validation. A method must be validated using
the protocols and reaction conditions under which it will be performed.
192. The Procedural Manual of the Codex Alimen tarius Commission, 12th edition, (2001), list the
main definitions of the parameters for method validation, although the concepts of a Limit of
Detection (LOD), Limit of Quantification (LOQ), and Range of Quantification (ROQ) are not yet
explicitly defined by Codex Alimentarius , they are listed below in order to answer this question:
193. Parameters for method validation: Accuracy – The closeness of agreement between the
reported result and the accepted reference value. A ccuracy describes how close the measured value is
to the actual value of a known reference sample.
194. Precision – The closeness of agreement between independent test results obtained under
stipulated conditions. Precision describes how well the results agree between repeated analyses of the
same material. Less precision is reflected by a larger standard deviation of the combined results than
the individual results.
195. Sensitivity – Change in the response divi ded by the corresponding change in the
concentration of a standard (calibration) curve, i.e., the slope of the analytical calibration curve.
196. Specificity – The ability of a method to respond ex clusively to the charact eristic or analyte.
Specificity describes how often the analyte is not detected if the analyte is not....

197. Ruggedness (Robustness) – The ability of a met hod to resist changes in results when
subjected to minor changes in environmental and procedural variables, laboratories, personnel, etc.
198. Applicability – The analytes, matrices, and concen trations for which a method of analysis is
validated that may be used satisfactorily.
199. Repeatability – Precision under repeatability conditions. These are conditions in which
independent test results are obtained with the sa me method on identical test items in the same
laboratory by the same operator using the same equipment within short intervals of time.
200. Reproducibility – Precision under reproducibility conditions. These are conditions in which
test results are obtained with the same method on id entical test items in different laboratories with
different equipment and operators.
201. Limit of Detection (LOD) – The lowest amount of analyte in a sample that can be detected
with suitable confidence but not necessarily quan tified as an exact value as determined by method
validation.
202. Limit of Quantification (LOQ) – The lowest am ount of analyte in a sample that can be
quantified with suitable accuracy and precision as determined by method validation.
203. Range of Quantification (ROQ) – The range within which the amount of analyte in a sample
can be quantified with suitable accuracy as determined by method validation.
 I will assume a PCR detection method for outline the necessary steps. According to ILSI 2001, there
are two primary ways in which PCR is used in th e detection of GM DNA in plants. These are called
quantitative PCR, which yields an estimate of the amount of the specific analyte present, and
qualitative PCR, which yields a yes/no answer as to the presence of GM material.
204. The Sample Preparation is considered the fi rst step and it is necessary to take into account
the:
• Sampling the consignment of seed or grain to obtain the bulk sample.
• Sampling of the bulk sample to obtain the laboratory sample.
• Subsampling the laboratory sample to obtain the test sample.
• Sampling the meal that results from grinding the test sample to obtain the analytical
sample.
• Sampling the DNA solution that results from extraction of the meal sample to obtain
the test portion.
205. The DNA extraction is the second step and DNA qu antification is the third. I could not find
more details for this topic. I also did not find re ferences for identify the differences in validation steps
for qualitative and quantitative detection methods but I found that the parameters common to
qualitative and quantitative methods are the specificity and applicability.
206. Reference Materials: According to ILSI 2001 , and ISO Guides 30 to 35, reference materials
play a number of roles in development, validati on and troubleshooting of PCR-based diagnostics, as
well as in the routine conduct of such assays. In the context of assay validation, positive reference
materials are used to establish the accuracy, precision, sensitivity, LOD, and the false negative rate in
quantitative assays. Negative reference materials are very important in determining the false positive
rates and specificity. At the ISO guidelines on reference materials, it is defined that they can be of
several levels of metrological quality:

207. A Certified or Standard Reference Material (CRM or SRM) is accompanied by a specific
certificate. This certificate states that one or more of the property values of the reference material is
certified by a procedure that establishes the value's tr aceability to an accurate realization of the unit in
which the property value is expressed; in addition, the certificate states a level of confidence of
uncertainty. Usually these reference materials are issued by National Metrology Institutes.
208. A Reference Material (RM) is a reference ma terial or substance one or more of whose
properties are sufficiently homogeneous and well estab lished to be used for the calibration of an
apparatus, the assessment of a measurement method, or for assigning values to materials. (ISO
definition).
209. A Working Standard (WS) is a secondary standard in regular use. This working standard is
equivalent to RM if it is quantified/characterized by comparison with the CRM/SRM. In order to
guarantee the quality standards for the reference ma terials, each one should be accompanied by a
certificate of analysis . The certificate will describe the characteristics of the material, both as to the
presence of the target material, and the absence of other possibly interfering materials. In addition, a
reference material may even be restricted as to th e method for which it can be used or is validated.
Usually, the certificate of analysis for a GM refe rence material will address the target event or
sequence; adventitious presence of other events; strength and purity; and genetic background. The
references consulted call the attention for the choice of the reference material, as they can have
advantages and disadvantages, according to particular purposes and usually there are three references
materials being used: Grain or seed or seed-deriv ed powders; GM DNA of plant origin; Plasmid
DNA or amplicons containing the target sequence. As I am not an expert in detection, I asked the
EMBRAPA technicians in this area and they pointed out that the key decision is the type of reference
material to use. This decision will be influenced by the availability of reference materials and must
consider the matrix effects. In any case, each method should be validated in the laboratory using a
reference material of the highest metrological standard available (SRM or CRM if possible). After
the validation, the laboratory may use a referenc e material or working standard that has been
calibrated back to the CRM/SRM.
42
210. Answer 8c: Qualitative methods indicate the pres ence or absence of the analytical target;
quantitative methods give a measure of the quantity of the analytical target present. For the purpose of
verifying compliance with thr esholds, both qualitativ e and quantitative DNA and protein based
methods can be used. It is essential to establis h appropriate sampling plans for both methods.
Statistical models that build the foundation of th e analysis of grain and seeds are made available
through ISTA and GIPSA/USDA.
211. An event-specific DNA-based method determines the presence of a specific trait at a specific
locus in a plant. A construct specific DNA-based method determines the presence of a gene within a

42 References:
International Standardization Organization, Geneva, Switzerland:
ISO Guide 30:1992 "Terms and definitions used in connection with reference materials",
ISO Guide 31:2000 "Reference materials -- Contents of certificates and labels",
ISO Guide 32:1997 "Calibration in analytical chemistry and use of certified reference materials",
ISO Guide 33:2000 "Uses of certified reference materials",
ISO Guide 34:2000 "General requirements for the competence of reference material producers ",
ISO Guide 35:1989 "Certification of reference materials -- General and statistical principles".
ILSI (2001) – Ilsi Europe Report Series: Method Development to in Relation to Regulatory
Requirements for the detection of GMO's in the food chain. Summary report of a joint workshop held in
December 2000. March 2001 (Prepared by the ILSI Euro pe Novel Food Task Force in collaboration with the
European Commission's Joint Research Centre (JRC) and ILSI International Food Biotechnology Committee)

plant, independent of its specific integration lo cus; a screening method determines the presence of
certain genetic elements, such as a promoter or a terminator. Protein based methods confirm the
absence or presence of specific expressed traits such as herbicide tolerance or insect protection
proteins.
212. Detection methods as applied to food or feed labeling do not have a direct role in risk
assessment or risk management. However, while similar methods are used to establish the level of
proteins in genetically modified products, and these levels are relevant to dietary intake and exposure
assessments that form the basis of risk assessment, the common understanding of "detection methods"
is that they are used to describe the presence or absence of GMO grain (or seed) in the context of
conventional grain (or seed). Thus, there are two ki nds of methods: methods that measure the analyte
in the grain for the purposes of characterization a nd risk assessment, and methods that detect the
presence or absence of the GMO (or level of the GMO) in seed, in grain or in a finished food or feed.
The manuscript referred to above describes the use of detection methods for this second purpose.
43
Question 9: In what ways does molecular charac terization inform the risk assessment for
any particular biotech product? Can a risk assessment be carried out in the absence of a
comprehensive molecular characterization of each transformation event?
Dr Nutti
213. Answer 9: According to the Codex Alimentarius Guidelines for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003, Paragraph 18), the
food safety assessment follows a stepwise process of addressing relevant factors. Molecular
characterization informs the risk assessment the st eps of the food safety assessment related to the
description of the recombinant-DNA plant, description of the genetic modification and
characterization of the genetic modification. Ho wever, all the safety assessment steps must be
fulfilled. My understanding is that the knowledg e achieved with a comprehensive molecular
characterization of a transformation event is fundamental for the food safety assessment of a biotech
product and, consequently, for its risk assessment.
214. The molecular characterization of a particular biotech product can be used to determine the
presence and absence of certain DNA-sequences. It can be further used to determine the integrity of
the inserted DNA, whether the sequence that was in tended to be introduced was indeed introduced
and whether it was introduced in a way that produces the expected protein products. Specific types of
molecular characterization could also be used to assess the amount of a certain DNA-sequence.
215. Sometimes, the term molecular characterization includes the biochemical characterization of
expressed proteins, and will include the description of the mode of action of a protein. Thus,
conclusions about the safety of an inserted DNA sequence and its expression product is dependent
upon: potential effects resulting from the assessment of the function of a gene, the mode of action a of
any encoded proteins; the relatedness of the inserted pr otein to proteins that have a history of safe use
or are related to protein toxins or allergens. This evaluation needs to incorporate the biochemistry of

43 References:
ILSI (2001) – Ilsi Europe Report Series: Method Development to in Relation to Regulatory
Requirements for the detection of GMO's in the food chain. Summary report of a joint workshop held
in December 2000. March 2001 (Prepared by the ILSI Europe Novel Food Task Force in collaboration
with the European Commission's Joint Research Centre (JRC) and ILSI International Food
Biotechnology Committee)

the organism the gene was introduced into, the organisms' intentional use, the conditions under which
the organism is cultivated, its interactions with the environment, and related factors.
Dr. Andow
216. A risk assessment cannot be carried out in the absence of a comp rehensive molecular
characterization of each transformation event. A risk assessment could not address the risks specific
to the transgene of interest without this information. Without the information, there would be no clear
basis for knowing when all reasonable scientific risk s have been assessed and there would often be an
uncertain basis for determining if a specific risk was scientifically justifiable or unjustifiable.
217. Molecular characterization addresses three distin ct issues: (1) specification of expected
expression of the transgene in the plant; (2) charact erization of expression of the transgene in the
plant; (3) genetic stability of the transgene under transmission to offspring.
218. Specification of the expected expression of the transgene in the plant is necessary to
characterize expression and to begin to clarify scientifically what can be considered an expected effect
and an unanticipated effect. Since about the year 2000, it has been technically routine and reliable to
determine the number of transgene loci, the copy nu mber at each locus, and of DNA sequence each
transgene locus. For some crops, most notably ma ize, it was also possible to sequence out from the
transgene through adjacent DNA until distinct plan t DNA sequences could be identified (these are the
so-called flanking regions). In 2001 this was not possible for some crops, such as cotton. Many other
molecular methods are under development, including proteomics and others, but these are not yet
ready for use in risk assessment now. It is likely that some of these tools will become available in the
future.44
219. Characterization of expression of the transgene in the plant is necessary to specify all gene
products from the transgene locus or loci and to determine their patterns of expression (concentrations
of gene products) in the plant, both as the plan t develops over time and as it generates new tissues and
plant parts. All gene products must be identified so that their effects can be assessed. If they are not
all identified, unanticipated effects could result fro m the unidentified products. It is important to
characterize patterns of expression because transgenes are typically expressed differently in different
plant tissues and expression in a plant tissue typically changes over time. In several cases, measured
differences among tissues and changes over time have proven significant for risk assessment.
220. Genetic stability of the transgene under transmission to offspring is needed to demonstrate the
transgene behaves similarly to ot her plant genes through development and reproduction. If it was not
stably inherited, most likely it would be silenced or excised or otherwise inactivated. In some such
cases, this could lead to unanticipated effects.
221. There is a paper that outlines the rationale fo r molecular characterization in detail for a
particular case.45

44 Nap, J.-P., A. Atanassov and W. J. Stiekema (eds). 2004. Genomics for Biosafety in Plant
Biotechnology. IOS Press: Amsterdam, 247 + viii pp.
45 Andow, D. A., D.A. Somers, N. Amugune, F. J. L. Aragão, K. Ghosh, S. Gudu, E. Magiri, W. J.
Moar, S. Njihia and E. Osir. 2004. Transgene locus structure and expression of Bt maize, pp. 83-116. In A.
Hilbeck and D.A. Andow (eds), Environmental Risk Assessment of Transgenic Organisms: A Case Study of Bt
Maize in Kenya. CABI: Wallingford, UK.

Dr. Healy
222. General Approach to Assessing Safety : It is well accepted that assessment of whole foods,
such as foods from genetically modified organism s (GMOs; GM foods), involves the use of the
'comparative approach', whereby GM foods are comp ared to their counterpart foods produced using
conventional techniques and which have been safely consumed over significant periods of time. The
objective of such a comparison is to identify any di fferences with the conventionally produced food.
Any identified differences are then examined for their biological impact in terms of health and safety.
223. This approach to the assessment of the safety of food products has been advocated for over a
decade (OECD, 1993,1996; WHO, 1991; 1996) and reconfirmed several times during that period.
Most notably, a relatively recent expert consu ltation convened by WHO/FAO concluded that 'a
comparative approach……is consider ed the most appropriate strate gy for the safety and nutritional
assessment of genetically modified foods' (WHO, 2000) . This approach is used in place of the
traditional approach of assessing the safety of discrete chemicals added to, or present in, food (eg food
additives, contaminants) that are generally subjected to a range of toxicological evaluations in animal
studies in a dose dependent manner (WHO, 1987). The toxicological approach is not appropriate for
whole foods because of the complex mixture of ch emicals present in a whole food, some of which
will be present at very low levels, and the difficu lty of conducting meaningful dose response studies
in animals while providing a nutritionally balanced diet.
224. The comparative approach to ex amining the safety of a GM food is generally considered to
encompass three major elements: molecular charact erisation of the inserted gene (transgene);
biochemical, structural and functional properties of the protein product from the inserted gene; and
analysis of the composition of the food. This approach taking into account knowledge about the
organism from which the transgene is sourced as we ll as the recipient organism. Application of the
comparative approach has been examined in de tail for food derived from GM plants (WHO, 2000;
WHO, 2001a), GM micro-organisms (WHO, 2001b) and GM animals (WHO, 2003).
225. The totality of information available from these types of studies should be considered in
coming to a view about the safety of the food; information from each element contributes to the
assessment of safety. Analyses of the information about these three aspects of the GM food provides
a structured mechanism to identify both the intended changes to the food as well as any that may be
an unintended consequence of the genetic manipul ation. Some unintende d consequences may be
predicted while others are unexpected.
226. The comparative approach involving characterisation at the DNA, protein and compositional
levels has been generally accepted and applied by many countries around the world, some for more
than a decade. These countries include Austra lia and New Zealand (ANZFA, 2001), Belgium (Van
Haver et al, 2003 ), Canada (Health Canada, 1994), European Union Member States (SCP, 1999;
EFSA, 2004), Japan (Japanese MHLW 2001), United Kingdom (UK FSA, 2003) and the United
States (USA FDA, 1992, 1997, 2001). Approaches by other countries are summarised in OECD,
2000 and Kuiper et al, 2001. A significant number of foods from GMOs have now been approved
using this assessment approach (eg Japan
46, Australia/New Zealand 47, Canada 48, USA 49 and
Europe50). International consensus on the appropriaten ess and utility of this approach resulted in the

46 http://www.mhlw.go.jp/english/topics/qa/dna/index.html.
47 http://www.foodstandards.gov.au/whatsinfood/gmfoods.
48 http://www.hc-sc.gc.ca/food.
49 http://www.cfsan.fda.gov/~lrd/biocon.html.
50 http://europa.eu.int/comm/food/food/biotechnology/authorisation/258-97-ec_authorised_en.pdf.

development of international guidance on the asse ssment procedures by the body responsible for
developing international food standards, the Codex Alimentarius Commission (CAC, 2003a, b, c, d).
Role of Molecular Characterisation in Risk Assessments
NOTE: QUESTION 9 REFERS TO THE ROLE OF MOLECULAR CHARACTERISATION IN A
'RISK' ASSESSMENT. HOWEVER, I HAVE ANSWERED THE QUESTION IN THE CONTEXT
OF A 'SAFETY' ASSESSMENT, WHICH IN RELATION TO EXAMINING THE HEALTH AND
SAFETY IMPACT OF FOOD, HAS A DEFINED MEANING (CAC 2003A) AND MAY BE PART
OF A SUBSEQUENT RISK ASSESSMENT IF POTENTIAL HAZARDS ARE IDENTIFIED
DURING THE SAFETY ASSESSMENT.
227. The molecular characterisation is considered to be a very important part of the safety
assessment of a biotech product. It provides in formation about the number of copies of the
transgene(s) that have been inserted, presence of any multiple copies at the same or dispersed
insertion site(s) and the structural organisation of the inserted transgene(s). The molecular
characterisation also includes an analysis of th e stability of the inserted DNA. Together this
information provides verification that the intended genetic modification has occurred and provides
some indication as to the likelihood of any un intended changes. However, the molecular
characterisation is not the only mechanism for id entifying unintended effects and the information
from such studies should be considered in conjuncti on with the information from the other elements
of the safety assessment.
228. While a safety assessment could be conducted in the absence of a comprehensive molecular
characterisation, current international good practi ce includes the molecular ch aracterisation (CAC b,
d). However, the issue does arise as to how comprehensive the molecular characterisation should be
in order to fulfil the requirements of an appropriate safety assessment. In large part, the answer to this
question depends on the type and quality of informati on available for the three elements of the safety
assessment, the information revealed from these st udies and the state of the scientific knowledge
about the gene(s) to be transferred, the donor organism and the recipient organism.
229. In recent years, there have been significant developments in the technologies to analyse the
molecular biological and biochemical characteris tics of GMOs and their products (eg DNA sequence
analysis, reverse transciptase polymerase chain reac tion, bioinformatics; micro array technologies,
proteomics and metabolomics). In some cases, these technologies can already significantly assist the
safety assessment. A recent example is the ready generation of DNA sequence data. It should be
noted that the guidelines for conducting safety assessments of GM foods that were recently adopted
by the Codex Alimentarius Commission (CAC 2003 b, d) require information sufficient to identify
any new substances that may be included in food. DNA sequence information on the junction regions
of the transgene and for the transgene itself is one mechanism to obtain this information.
230. It is now regarded as good practice to undertake DNA sequence analysis as part of a safety
assessment, for several reasons. First, some of the gene constructs and the transformation
technologies available today result in more complex segments of DNA and in rearrangements of the
inserted DNA. One of the most effective methods currently available for assessing the integrity and
impact of the inserted DNA is sequencing. Second, sequence information can now be much more
readily generated than was possible using the laborious procedures available a decade ago.
231. Although this type information has not genera lly been required in the past, a number of
countries have now included requirements for its pr ovision in the future. However, countries have

generally not required such information for GMOs a nd their food products that have previously been
approved for entry to the marketplace.
232. Some of the technologies being developed are currently not appropriate for use in safety
assessments as they do not yet provide any further assu rance of safety or the results cannot be fully
interpreted (eg NAS, 2004). However, some of the profiling methods, such as metabolomics, may
eventually prove very useful. At some point in the future, the development of these technologies and
associated understanding about the impact of sma ll variations in gene expression levels and/or
metabolites may be adequate, minimising the need for molecular characterisation.
233. It must also be acknowledged that the commerc ially produced GMOs gene rated to date have
utilised well defined crops as recipients of the gene tic modification and well defined genetic elements
in the transgenes. Generally, the genetic elements have been extensively studied at both the DNA and
protein levels. Consequently there has been a high level of confidence in the assessments carried out
to date. However, there have been ongoing develo pments in the molecular biological technologies
that are being applied in the development of GM crops (eg different transformation technologies;
insertion of multiple and different genes; differe nt donor and recipient organisms) and these are
raising new and more complex questions about the saf ety of the products of such crops in the food
supply.
234. Conclusion: Currently the molecular characterisation is an important element of the safety
assessment, providing verification that the intended genetic modification has occurred and
information about unintended effects. The extent of molecular characterisation in any particular case
should be addressed in the context of the available scientific information.
235. Approach to Addressing Specific Questions About Molecular Characterisation :
Questions relating to requests for additional information for safety assessment purposes have been
considered in the context of the general approach to assessing the safety GMOs and their food
products. In particular, requests for further molecular characterisation have been considered
according to the principles and approaches outlined above.
Dr. Squire
236. The Guideline for the conduct of food safet y assessment of foods derived from recombinant-
DNA plants indicate full characterisation of the gene tic change should be provided. Such information
may be essential for assessing some aspects of th e product but is not always necessary when
considering spread or persistence of the biotech product or its effect on the ecosystem.
237. Spread and persistence. In many of the studies in Europe (ongoing and not yet published), the
purposes of the study can be achieved by detection based on an outward or phenotypic trait (i.e.
whether a plant dies if sprayed with a specific herbicid e), a protein test (e.g. an antibody based test for
the protein produced by the genes conferring tolera nce to glyphosate or glufosinate ammonium) or a
DNA test for the gene (e.g. the bar gene). None of these tests is absolutely accurate since they are
affected by the condition of the plant tissue. If samp ling of plant material is done in real agricultural
fields, it is impossible to ensure the plant tissu e is in the right condition – often it is not.
Consequently, false-positives and false-negatives occur. Moreover, if the GM trait is thought to be at
low frequency, e.g. 0.5% or lower, and the individuals possessing it are unevenly distributed or
clumped in the field, then very large numbers of individuals have to be sampled to estimate presence
or % frequency with a high degree of certainty.

238. Ecological impact. For some aspects of comparing GM and non-GM plants, detailed
knowledge of the molecular construc t is not necessary. For instance, if comparing the ecological
effects of GMHT or Bt plants, the studies coul d be done equally well wi thout detailed molecular
knowledge of the construct.
239. More information would be necessary if r esearch was following the volunteers or pollen-
vectored genes originating from the GM crop: at least, knowledge of the genetic factors determining
the proportions of male sterility and male fertility would be important, but even then the detailed
molecular knowledge of the construct would not be essential. However, if many GM varieties had
been grown in an agricultural area, and if it were important to ascertain the origin of an individual
plant or population, then more detailed molecular characterisation would be required.
240. More generally it can be argued, given the scientific and public interest in GM issues, that it is
reasonable for an assessing body to request full ch aracterisation (according to standards in the
Guideline for the conduct…) of a GM construct (a) to enable its officers to confirm whether
impurities existed in the original seed if they we re found or suspected (e.g. the presence in the
breeding line of GM traits such as antibiotic resistance) and (b) to trace the persistence or spread of
the GM trait if this was necessary in later years.
241. While much of the text above under Responses 9 is generally appreciated by workers in the
field, many of the scientific findings pertaining to th e spread and persistence of GM traits in Europe
are not yet in the public domain (Notes, paragraph 14).
ISSUE 1

Background: The following questions have the objective of assisting the Panel to determine whether
the EC regulatory approval process for the specific biotech products at issue in this dispute
proceeded without unjustified delays on the part of the relevant EC regulatory bodies. The questions
are aimed at determining for each product application (whether pending or withdrawn):

(a) the scientific or technical grounds for the comments and/or objections raised by EC
member States following the initial assessments of the Competent Authority (CA) and of the
relevant EC scientific body, in light of the sci entific evidence and evaluations available at the
time;

(b) the scientific or technical grounds for the requests for additional information from
the notifier during or following the initial a ssessments of the CA and of the relevant EC
scientific body, in light of the scientific evidence and evaluations available at the time; and

(c) the justification on scientific or technica l grounds for the time taken to evaluate the
additional information provided.

Questions:

Bayer oilseed rape (Falcon GS40/90)
C/DE/96/05 (EC chronology 62)

Question 10: Given the information before the Panel, including the notification by AgrEvo (EC-
62/At.1-30) and the EC Scientific Committee for Plant (SCP)'s opinion (EC-62/At.74), was the
information to assess the long-term effect of th e newly expressed protein on the biogeochemical

cycle and the food chain requested by the Ita lian CA (EC-62/At.95) necessary to ensure that
conclusions of the safety assessment were valid?
Dr. Nutti
242. Based on the information provided by the appli cant and the analysis carried out by the EC
Scientific Committee for Plant (SCP) and following the Codex Guidelines, my understanding is that
the request by the Italian CA (EC-62/At.95) was not necessary to ensure that conclusions for the
safety assessment of the newly expressed protein in the food chain were valid. As pointed out by the
EC SCP, the new protein has a very low potential for being allergenic or toxic and also is degraded
within the gastric fluids. In my opinion, the conc lusion of the EC SCP that the protein was safe for
consumption in the food chain is correct and w as based on sound scientific evidence, which was
presented by the applicant.
Dr. Andow
243. The request by the Italian CA (EC-62/At.95, 14 March 2000) is repeated below.
Referring to the conclusion of the last meeting of the Regulatory Committee under
Directive 90/220/EEC, Italy would be very pleased to receive from the notifier more
information concerning the assessment of the effect of the genic product on the
biogeochemical cycles and on food chain and on the apreading of the gene due to the
possibility of crossing between the PGM and wild species.
244. The original notification by AgrEvo (EC-62/At.1-30, 1 April 1996) contains no studies on the
safety of Falcon GS 40/90 to other organisms and no studies on outcrossing rates with wild species.
Instead, the notifier argues that Falcon GS 40/90 is in no way different from non-transgenic cultivars
expect for the ppt gene. They conclude that ther e is no reason to believe that Falcon GS 40/90 has
any adverse effects on the environment.
245. In my reading of the original notification, AgrEvo's claim rests entirely on the SCI report
"Glufosinate tolerant transgenic Winter Oil Seed Rape – Field observations of the variety Falcon GS
40/90 pHoe6/Ac in comparison to Falcon non transgenic and conventional fa rmers rape," which is
code number Hoe 039866 00 SL 01 A 201 (EC-62/At.30_SCI, 1 April 1996), and on the report
"Supplementary ecological monitoring of a transgenic , Basta-resistant winter rapeseed crop cultivated
at Friemar (Thuringia, Federal Republic of Germ any) during the season 1994/95" (EC-62/At.8). I
agree with the assessment of the lead CA that EC-62/ At.8 is difficult to interpret. Without going into
the details, the data are not scientifically convin cing that the environmental effects of Falcon GS
40/90 are in no way different from non-transgenic cultivars.
246. The lead CA requested improvements to EC-62/At.8 on 14 May 1996 (EC-62/At.33) and
received an improved version from the notifier on 12 July 1996 (EC-62/At.37_SCI). This includes a
revision of EC-62/At.8. The notifier admits that the small plot size in EC-62/At.8 precludes statistical
analysis, so I conclude that the data are of li mited scientific value for evaluating environmental
effects.
247. The full application to the community was not included in documents before the Panel, so it is
not possible to determine conclusively the basis for the SCP assessment.
248. Between the time of submission of the full appli cation to the community and the time of the
SCP Opinion, there were several requests for information to the notifier from various countries and

the SCP. The replies from the notifier are in EC- 62/At.45, EC-62/At.52, EC-62/At.54, EC-62/At.68
and EC-62/At.71. Information in EC-62/At.52 and EC-62/At.71 are relevant to the question at hand.
249. In EC-62/At.52, 28 January 1997, the notifier clarified as follows:
42. Survival, multiplication, and dissemination of the GMO(s) in the
environment:
The comparison study mentioned in this section has only taken place in a growth
chamber which is big enough to handle ev en maize plants. Field observations are
discussed in document A 56991.
Five plants each from the recipient and from the transformant have been analysed in
the growth chamber.
250. In EC-62/At.71, 5 June 1998, the notifier provided extensive information related to
environmental impact that was not available previo usly. This addressed dispersal of oilseed rape
pollen, the potential invasiveness 51 of Falcon GS 40/90 oilseed rape, and a number of aspects related
to fitness, weediness and outcrossing (data and sources not provided).
251. The SCP concluded that there is no evidence to indicate that Falcon GS 40/90 is likely to
cause adverse effect on human health and on th e environment (EC-62/At.74, 14 July 1998).
However, they also indicate that "Few studies have been conducted on the safety of modified oilseed
rape to other organisms." No studies were conducted on biogeochemical cycling or on the food chain.
252. It must be stated clearly at this point that th e lack of evidence does not imply that there is a
lack of an effect. Either the SCP did not base their conclusion about non-target effects on sound
scientific reasoning or the reasoning they used is no t apparent from their Opinion (EC-62/At.74). In
either event, their conclusion does not follow from the data provided by the notifier.
253. The SCP did address the possibility of the spr eading of the gene due to crossing between
Falcon GS 40/90 and wild species. The SCP indicated that the likelihood of crossing to wild species
was small, especially when compared to the likelihood of crossing with another oilseed rape cultivar.
They suggested that appropriate risk management measures would be likely to avoid these risks.
254. The conclusions by the SCP related to outcr ossing and invasiveness are based on the sound
interpretation of the data present at the time. Howeve r, there is likely to be several approaches to risk
management, and these are not specified.
255. The Italian CA (EC-62/At.95) requested inform ation to assess the long-term effect of the
newly expressed protein on the biogeochemical cy cle and the food chain. While a request for
additional/ clearer assessment of the possible effect s on environment can be readily justified on the
basis of the preceding analysis, such a request must it self be clear and specific. For example, it is not
clear that all of the environmental hazards have been identified let alone assessed. A request for
additional/ clearer assessment of the possible effects on the environment would not be clear or
specific enough to allow a notifier to know how to respond, and hence although information on the
environmental impacts is necessary to ensure that conclusions of the safety assessment were valid, a
general request for such an assessment would not be an appropriate way to request such information.
In a similar way the request by the Italian CA is not clear or specific enough to be considered

51 Crawley et al. Nature 363: 620-622.

necessary. There are hundreds of possible processes in biogeochemical cycles that could be
investigated, and thousands of possible ways to evaluate food chains. Unless the Italian CA can point
to prior regulatory precedents wh ere these terms are clarified, the phrasing of the request is not
scientifically justified. Alternatively, had the It alian CA specified certain biogeochemical cycles and
food chains to assess, then it would be possible bo th for the notifier to assess them and for me to
determine their necessity to ensure that conclusions of the safety assessment were valid.
256. The Italian CA request came on 14 March 2000 . On 10 November 1999 Aventis submitted
and updated notification, which was circulated to the CAs on 13 December 1999. The AFFSA
Opinion was issued 5 January 2000 and Aventis provided additional information on 28 February
2000. The time between the receipt of materials un til the Italian CA response can be calculated as
either 15 or 125 days. Of the 125 days, 33 were used to process the file administratively, and Italy
had the materials for 92 days. The shorter time is a very rapid response time. The longer time is
probably too long, but mainly because 33 da ys seems excessive for processing the file
administratively.
Question 11: Given the information before the Pa nel, including the notification by AgrEvo and
the conclusions of the EC SCP, was the info rmation regarding the post-event monitoring
protocol to assess the likelihood of spreading re quested by the lead CA necessary to ensure that
conclusions of the safety assessment were val id (EC-62/At.100)? If so, what other risk
management options are available to mitigate this risk?
Dr. Nutti
257. Three questions presented in EC-62/At.100 are on environmental issues. I can only refer to
item 7.3 (oilseed rape animal feeding study), where a doubt was raised about which event was used in
the male broiler chicken studies. It is mentioned in EC-62/At.103 that copies of both studies were
submitted by the notifier in order to answer this point. I could not find these studies in the
attachments, but examining the information provided by the applicant to EC and the answers given in
EC 62 (Attachments 003, 005, 006, 007, 010, 011, 012, 015, 017, 018, 019, 021, 022, 023, 024, 025,
026, 027, 028, 049, 050, 051, 053, 057, 060, 063, 065, 071, 073, 074, 075, 081, 082, 085, 091, 093
and 0103) and the analysis carried out by the EC Scientific Committee for Plant (SCP) (Exhibit US-
42), where it is relevant to note:
258. Item 7 (Overall Assessment): "there is no evidence to indicate that the placing on the market
of line transformant Falcon GS 40/90 oilseed rape, with the purpose to be used as any other oilseed
rape is likely to cause adverse effects on human health and on the environment".
259. Item 6.2.2: "in food of animal origin from liv estock animals fed with feedingstuffs after the
application of glufosinate-ammonium to tolerant rape, no residues are expected above the limit of
determination".
260. My understanding is that there is no significant risk to humans or livestock with the ingestion
of the gene product. All the food safety aspects indi cated in the Codex Alimentarius Guidelines have
been accomplished. It is important to point out that the nutritional composition of the entire seed,
meal and oil from the genetically modified rape has been analysed. Information on the contents of oil,
fibre, ash, protein, fatty acid composition, glucosinolate, sterols and tocopherols showed no
differences that could be attributed to the genetic modification. The Scientific Committee for Plants
appointed within the EC Commission has, in its opinion on rape, stated that no significant changes in
nutritional value have been identified.

261. I cannot express any opinion on the post-event monitoring protocol since this is an
environmental issue. According to the follow- up programme, monitoring will be carried out on
herbicide use, biodiversity and spread of the genes introduced via waste plants and wild relatives.
Dr. Andow
262. The information requested by the lead CA rega rding the post-event monitoring protocol to
assess the likelihood of spreading is in EC-62/At.100, 25 July 2001, and reproduced below.
Ad_Document_3 ("Draft Post-Marketing Monitoring Plan for InVigor ®, SeedLink®
and LibertyLink® Oilseed Rape"):
III.2. "Existing networks of expertise to carry out General Surveillance":
On principle, we think that a more deta iled description of the tasks and the kind of
data that will be collected by the different networks would further acceptance of the
monitoring plan by the other Member States. We expect that relevant information
will be supplied by Aventis after the diffe rent networks have elaborated their
programs for the general surveillance.
III.2.2. "Plant protection networks":
In Germany, the "Bundessortenamt" is th e competent authority for plant variety
registrations (as mentioned under III.2.1.), but its focus is not on evaluating the
impact of agricultural practices on the environment, specifically on water, soil,
wildlife and flora. Since several federal and regional ("Länder") authorities are
responsible for these questions in Germany, it would be difficult to give a complete
list. We suggest either to omit reference to a particular institution in Germany or,
alternatively, to mention – as the main federal and regional institutions – the
"Biologische Bundesanstalt für Land- und Forstwirtschaft", the "Umweltbundesamt"
and the "Pflanzenschutzämter".
263. The first mention of a monitoring protocol is in the SCP Opinion (EC-62/At.74, 14 July
1998).
2. The SCP is also of the opinion that the potential transfer of the herbicide resistance
gene to wild Brassica relatives is a new issue in Europe in view of the limited scale of
release to date. The SCP has examined the available evidence from monitoring and
research programmes to date. After evaluating all the information available to the
SCP, it was concluded that herbicide tolera nt volunteers that may appear would be
canola plants and not wild Brassica relativ es. Such herbicide-tolerant volunteers
could be controlled in subsequent crops by conventional agricultural methods other
than by the use of glufosinate-amm onium. The SCP recommends that the
introduction of herbicide tolerant crops should be accompanied by:
i) an agreed code of practice for field ma nagement of the particular modified crop
involving the active participation of the notifier to promote best practice by farmers.
ii) a research programme with an agreed design and implementation plan to detect
the occurrence and the establishment of herbicide tolerant volunteers and weeds
under field conditions in the EU.

264. The SCP is recommending a "code of practice" in order to ensure that herbicide-tolerant
volunteers are eliminated. In other words, the purpose of the risk management measure is to eliminate
volunteers as they occur. In addition, it is re commending research design and implement monitoring
to detect herbicide-tolerant volunteers and herbicide-tolerant weeds in the EU.
265. The notifier indicates they are developing a monitoring plan in EC-62/At.75, 22 June 1998,
even before the SCP is published), with some deta ils in Annex 3. Annex 3 is not a part of the
materials before the Panel.
4. that PGS and AgrEvo contribute to and collaborate with different programmes
monitoring the occurrence and establishment of herbicide tolerant volunteers and
weeds under field conditions. In annex we present examples of collaborations in the
UK, France and Germany. PGS and AgrEvo will keep the Scientific Committee on
Plants as well as the competent authorities informed of the results.
266. The notifier provided a monitoring plan (EC62/At.78, 25 January 1999) entitled "A
Structured Stewardship and Monitoring Programme for Agronomic Impact of Growing LibertyLink™
and SeedLink™ Oilseed Rape in Europe – A mon itoring and quality assurance strategy." The main
objective of this plan is to increase understanding of the appropriate management of these products by
their users and to improve public awareness and acceptance. 52 The second objective is to promote
practices, which ensure the quality and stability of the crops, in both the farmers' and AgrEvo's own
interest.53
267. In the monitoring plan is a section entitled "2 .3.2 Which Guidance?" in which the notifier
states that they will provide farmers with guidan ce on volunteer management: post-harvest treatment
of the area. However, there are no details on the gui dance that will be provided. Moreover, there is
no concrete discussion on how a monitoring plan will be implemented (see my answer to question 24a
for criteria on judging the completeness of a monitoring plan).
268. This monitoring plan (EC62/At.78, 25 January 1999) does not meet the specifications
suggested by the SCP.
269. In EC-62/At.81-82, 8 July 1999, the notifier repeats information in EC-62/At.71, 5 June 1998.
270. In EC-62/At.83, 3 August 1999, the notifier reiterates the content of EC-62/At.78, 25 January
1999.
271. In EC-62/At.90_SCI, 10 November 1999, the notifier modifies the proposed monitoring plan
from that in EC62/At.78, 25 January 1999. The main objective is still to increase understanding of
the appropriate management of these products by their users and to improve public awareness and
acceptance. There is no secondary objective. [xxx]
272. The lead CA requested (III.2 in EC-62/At.100, 25 July 2001) more detailed description of the
tasks and the kind of data that will be collected by the different networks. As the monitoring
objective of the notifier does not fully correspond wi th the monitoring recommendations of the SCP,
or monitoring objectives of what will become Annex VII of Directive 2001/18/EC, and there are few
details in any of the proposed monitoring plans submitted by the notifier, and the submitted
monitoring plans do not address the main manage ment measure recommended by the SCP (how

52 EC-62/At.78, p. 9 of pdf file.
53 EC-62/At.78, p. 9 of pdf file.

conventional practices will eliminate volunteers), th is request for additional information is necessary
to ensure that conclusions of the safety assessment were valid.
273. The lead CA also requested (III.2.2 in EC-62/At.100, 25 July 2001) a modification to the list
of proposed plant protection networks that will participate in monitoring. This seems like a useful and
important suggestion to the notifier, and should be c onsidered necessary to ensure that conclusions of
the safety assessment were valid.
274. There are many other possible risk management options to the monitoring systems proposed
by the notifier. The most evident is to follow up rigorously on the SCPs original suggestion that
conventional practices may be sufficient to eliminat e volunteers. This would require the notifier to
consider all the possible practices that may follow an oilseed rape crop and determine which aspects
of these are responsible for elimination of volunteers. I would suggest that the competitiveness of the
subsequent crop, the phenology of crop management activities (disturbance regime), and management
of field margins (tillage or herbicide application) may be critical factors that would be sufficient to
eliminate nearly all volunteers. Monitoring would shift from looking for glufosinate-tolerant
volunteers and weeds to monitoring the cropping prac tices of farmers. Monitoring for glufosinate-
tolerant volunteers and weeds could then be restricted to those farmers or regions where management
of the subsequent crop was inappropriate for eliminating volunteers.
275. The lead Ca requested the information on 25 July 2001. The notifier provided the reviewed
proposal on 10 November 1999. This is a 1 year 8 month interval, and is excessively long.
Dr. Snow
276. In summary, asking for information about the post-event monitoring protocol appears to be
justified for two reasons. First, when the EC SC P rendered its opinion in July 1998, questions were
raised about how quickly transgenic resistance to glufosinate would spread to volunteer oilseed rape
plants and to weedy Brassica rapa. The EC SCP stated that these questions could be addressed by
post-release monitoring. Second, information about monitoring is now required under EC Directive
2001/18, which went into effect in March 2001. Discussions about whether to require monitoring
protocols were probably taking place while the previous Directive was in effect as well.
277. With regard risk management options, the spr ead of volunteer oilseed rape or wild Brassica
species that acquire resi stance to glufosinate can be managed by other methods of weed control,
including applications of other herbicides. However, if glyphosate-tolerant oilseed rape is introduced
in the future, problems with herb icide-resistant volunteers could be compounded. This concern is
mentioned specifically in the EC SCP (EC-062-At. 074).
278. I noticed two shortcomings in the notifica tion for Falcon GS40/90 by AgrEvo (EC-062-At.
003) and the July 1998 conclusions of the EC Scientific Committee on Plants (SCP; EC-062-At. 074):
(a) The documents show that both AgrEvo and the EC SCP concluded that transgenes
from oilseed rape are unlikely to spread to weedy Brassica rapa and multiply, despite
the fact that several early studies showed that persistence of the transgenes is
expected (Jeorgensen and Andersen 1994, Mikkelsen et al. 1996, Snow et al. 1998).
The SCP stated ". . . there may be a very low level of natural crossing with related
species particularly Brassica rapa and Brassica juncea under field conditions. . . The
risk assessment assumes that transfer will occu r at a very low level." This could be
correct, depending on local conditions. Ho wever, the EC SCP then stated: "After
evaluating all the information available to the SCP, it was concluded that herbicide

tolerant volunteers that may appear would be canola plants and not wild Brassica
relatives." The basis for this conclusion is not clear (also, it doesn't make sense to
refer to wild relatives as a type of volunteer).
(b) AgrEvo and the EC SCP did not specifically address the issue of how the widespread
adoption of glufosinate-tolerant canola would affect the efficiency of weed control,
which has implications for biodiversity in farmland habitats (see above). This
question had not been evaluated at the time and is still under investigation.
Dr. Squire
279. If long term events or emergent properties are envisaged as a result of introducing a GM plant
and practice (and this seems to be the case for oilseed rape in Europe), then it is legitimate to expect a
rigorous post-event monitoring protocol to be integral to a pre-release risk assessment. Risk
management options for this GMHT product are si milar to the generic ones included in Responses
6d-e. When they are applied, the practices appear to work in keeping weedy oilseed rape to
manageable levels, but the gene ral level and type of management have not prevented it from
becoming established throughout Europe as a weed.
Question 12: Given the information before the Pa nel, including the notification by AgrEvo and
the conclusions of the EC SCP, was the information regarding the molecular characterization of
this product requested by the lead CA (EC-62/At.106) necessary to ensure that conclusions of
the safety assessment were valid?
Dr. Healy
280. In 1996 AgrEvo submitted a request to the German CA for the authorisation of the marketing
of oilseed rape carrying the Falcon 40/90 tran sformation event. The German CA expressed a
favourable opinion in late 1996. Subsequently , the request was forwarded to the European
Commission and distributed to the Member States in December 1996. A number of the Member
States raised concerns and requested that additiona l information be provided. The original dossier,
together with additional material supplied, was considered by the Scientific Committee for Plants
(SCP), with a favourable opinion being issued in mid 1998.
281. Information Submitted (EC 62 – 1-30 37, 73): The recipient oilseed rape line was
transformed with a gene encoding the enzyme phosphinithricin acetyl transferase (PAT) from a
common soil bacterium. The PAT enzyme inactiv ates the herbicide glufosinate ammonium and
confers herbicide tolerance to the genetically modified oilseed rape. The gene was constructed so that
the regulatory sequences for the pat coding region were derived from the cauliflower mosaic virus.
These genetic elements are well described in the litera ture and have been used to transform a variety
of crops, both in the combination used in this case as well as in combination with other coding regions
and/or regulatory sequences. Furthermore, a numbe r of oil seed rape lines transformed with the pat
gene, sometimes in combination with other genes, have been assessed for safety and approved for
food use in a number of countries around the world (eg Japan
54, Australia/New Zealand 55, Canada56,
USA57 and Europe58).

54 http://www.mhlw.go.jp/english/topics/qa/dna/index.html.
55 http://www.foodstandards.gov.au/whatsinfood/gmfoods.
56 http://www.hc-sc.gc.ca/food.
57 http://www.cfsan.fda.gov/~lrd/biocon.html.

282. The molecular characterisation submitted included a sequence analysis of the pat gene
isolated from the soil bacterium, full DNA sequence analysis of the DNA construct containing the pat
gene and Southern blot and Polymerase Chain Reac tion (PCR) analysis of the transgene. The latter
demonstrated that the pat gene was stably inserte d into two separate locations within the recipient
strain and that vector sequences outside the border regions had not integrated. Analysis of the
biochemical and structural properties of the protein included heat stability, pH optimum, enzyme
kinetics, inactivation studies in th e presence of gastric juices from a variety of species and under a
variety of conditions. These studies were conducted with the PAT protein isolated from a variety of
transformed organisms, including an over expressing E. coli strain and maize and oilseed rape line
transformed with the pat gene, which were demonstr ated to have similar immunological properties to
each other. The DNA sequence of the pat gene used to generate Falcon GS 40/90 was compared to
major sequence databases. These studies also allow an assessment of the allergenicity potential of the
PAT protein.
283. The PAT protein was tested for dose responsive toxicity effects in animals. The treated
animals were subjected to extensive analysis for toxic effects including mortality, food consumption,
clinical parameters, clinical biochemistry and pathology on an array of internal organs.
284. The composition of fractions derived from seeds we re analysed for nutrients appropriate for
the food (oil) and feed derivatives of oilseed rape (OECD, 2001). The analysis addressed the key
fatty acids in oil (including the toxin erucic acid) as well as nutrients and toxins in oilseed rape meal.
285. Conclusions of the Safety Assessment: The SCP in July 1998 (EC 62-74) concluded that
having considered the submitted dossier and the background information available, 'there is no
evidence to indicate that the placing on the market of line transformant Falcon GS40/90 oilseed rape,
with the purpose to be used as an y other oilseed rape, is likely to cause adverse effects on human
health….'. Furthermore, the SCP did not request any additional information or suggest any additional
research in relation to the safety assessment. It appears that the information submitted met the
requirements for a safety assessment as outlined in Annex 11 B of Directive 90//220/EEC.
286. I have reviewed the scientific information submitted and consider that a reasonably
comprehensive information package was developed to support the safety assessment of oilseed rape
line Falcon GS 40/90. The information package de veloped was consistent with internationally
accepted approaches to the safety assessment at that time, as outlin ed above. The information
package addressed the three major elements that are usually cons idered in assessing the safety of a
GM product and included molecular characterisation of the inserted gene, characterisation of the
protein expressed by the inserted gene and an analysis of key compositional constituents relevant to
the food product (ie oil) derived from the Falcon GS 40/90 line. The experimental procedures were
conducted in accordance with standard methodologies.
287. Having examined the scientific information underpinning the safety assessment and the
opinion of the SCP, I conclude that food (oil) de rived from oilseed rape Fa lcon GS 40/90 line is as
safe for human consumption as food from conventional produced oilseed rape varieties. There is no
indication from the scientific evidence submitted for consideration by the Competent Authority that
the food poses a health or safety risk. Furthermore, there is no indication of human health and safety
issues arising from the consumption of food from other lines of oilseed rape carrying the pat gene that
have been approved around the world and which ar e/have been in commercial production, some for
significant periods of time. I therefore concur with the opinion of the SCP, issued in July 1998.

58 http://europa.eu.int/comm/food/food/biotechnology/authorisation/258-97-ec_authorised_en.pdf.

288. The Lead CA sought addition information (EC 62-106):
• on the DNA sequences flanking the transgene locus in Falcon GS 40/90 (ie spanning
the left and right junctions between the inserted DNA and the plant DNA),
• comparison of these DNA sequences with those of the pre insertion site in
nontransformed lines, preferably from the Falcon variety.
289. The information on the flanking sequences is lik ely to provide a full se quence analysis across
DNA junction between the genomic DNA and the inserted DNA (pat gene). Such information can be
used to analyse the DNA for potential novel protei ns that may be generated as a result of the
juxtapositioning of the different segments of DNA.
290. Such information will also provide a more detaile d description of parts of the structure of the
transgene that was actually inserted into the genom ic DNA, facilitating the identification of any small
rearrangements or other modifications to the DNA th at may occur during the transformation process.
Some transformation techniques are more prone than others to generating such modifications.
291. Comparison of the insertion sites in the transformed line with that of its parental non
transformed line may give additional information about any minor DNA changes at these sites (eg
rearrangements, deletions). It may also reveal other nucleotide changes (polymorphisms) that are a
result of natural variation between individuals within lines and which are not related to the
transformation process.
292. Impact of the Additional Information on the C onclusions of the Safety Assessment: The
additional information requested by the Lead CA is unlikely to influence the conclusions of the safety
assessment carried out by the SCP. The rationale for this view are:
• The transgene was inserted into the Falcon GS40/90 line via agrobacterium mediated
transformation techniques. This technique is less likely to result in DNA
rearrangements and other modifications of the inserted DNA, compared to some other
transformation techniques (eg micro projectile bombardment). The molecular
characterisation provided by AgrEvo does not indicate that any modifications to the
structure of the inserted DNA have occurred although the possibility of small
modifications that cannot be detected by Southern blot analysis cannot be entirely
excluded. This is well exemplified by the example of Roundup Ready soybean, for
which the initial molecular ch aracterisation did not reveal any modifications in and
around the inserted gene, although subseque nt sequence analysis revealed the
presence of additional DNA fragments (Windels et al, 2001).
• The molecular characterisation originally submitted did include a PCR analysis,
which indicated that only the DNA sequences associated with the pat gene had
integrated into the recipient strain. In pa rticular, the analysis verifies that that no
sequences from the vector, others than intended, integrated into the recipient strain.
• Additional information of the sequences at the sites of insertion may reveal fine
structural details and identify similarities and differences with the parental line at
these sites. Such detailed structural analyses usually aim to provide an indicator of
unintended effects from the genetic modification. However, in some cases it may be
difficult to determine the biological signif icance of any sequence differences because
of a lack of knowledge about the extent of sequence variation between individual
organisms within the parental line. Furthermore, other indicators of unintended
effects, that are biologically meaningful, are obtained from other elements of the
safety assessment (eg analysis of protein, composition of food).

• A comprehensive package of informati on was provided that addressed the key
elements required in a safety assessment and there was no indication of any safety
issues when all the available data was take n into account. Furthermore, the pat/PAT
gene/protein is well characterised, has been utilised successfully in a number of crops
and their resultant food products and was derived from a well described organism.
293. Conclusion: The additional molecular characterisation requested by the Lead CA is unlikely
to affect the conclusions of the safety assessment and was not necessary to ensure that the conclusions
of the safety assessment were valid. The totality of information provided in the dossier to that point in
time were adequate to support the validity of the conclusions of the safety assessment without the
need for the additional data requested in the letter dated 2 April 2002.
Dr. Snape
294. The comments below relate to Question 12 concerning the molecular characterisation of
exhibit EC-062. These were examined relative to the EFSA-Q-2003-005 guidance document
produced by the Scientific Panel on Genetically Modified Organisms.
295. Exhibit EC-062: Oilseed rape event Falc on GS40/90 (also named GS40/90pHoe6/Ac) was
produced, using plasmid pHoe6/Ac, by means of Agrobacterium-mediated gene transfer. The T-DNA
of the plasmid pHoe6/Ac used for transformation carries a chimeric pat expression unit conferring
resistance to the herbicide glufosinate. Oilseed rape event GS40/90pHoe6/Ac contains two functional
transgenic loci. Both transgenic loci will be present in the commercialised GM plants. Information
relative to the molecular characterisation of GS40 /90pHoe6/Ac has been found primarily in EC-062
attachments 3, 13, 14, 113, 114. A series of commen ts and questions are numbered below. This is
followed by a conclusion proposing answers to Question 12.
296. The molecular characterisation of GS40/90pHoe6/ Ac initially provided was not complete and
failed to identify the exact transgenic make-up of th is GM plant. Initially, two intact copies of the
CaMV35S:pat:35T expression unit were believed to be inserted at tw o independent f unctional loci
(EC062 attachment 3 and 14). Later it was determin ed, following requests for additional information,
that in fact one of the two functional loci contained two CaMV35S:pa t:35T expression units in
inverted tail-to-tail orientation (EC062 attachment 114) bringing the total copy number of the pat
gene to three in GS40/90pHoe6 /Ac. This re-emphasizes the need, expressed in general comment 2
(accompanying document) by this Expert, to conduct independent molecular analyses for each
transgenic locus present in a GM plant. This also means that two sets of full molecular analyses are
expected to be provided, one for each transgenic locus present in GS40/90pHoe6/Ac.
297. The segregation analysis of transgene phenotyp e indicates the presence of two functional loci
in GS40/90pHoe6/Ac but does not account for n on-functional transgenic sequences potentially
inserted into additional genomic location (i.e. third transgenic locus). An inheritance study, based on
the presence or structure of transgenic sequences in progeny plants should be conducted to confirm
that only two transgenic loci are present in GS40/90pHoe6/Ac (see comment 3 in accompanying
document).
298. EC062 – Attachment 3: The text cross-refere nce annexes that are not included in this
attachment. This reviewer assumed that annex 8 and 9 corresponded to attachment 13 and 14.
299. EC062 – Attachment 13: Only a partial sequenc e of the pHoe6/Ac vector (including the T-
DNA region and surrounding vector sequences) is provided. Attachment 13 (annex 8) does not
provide information about the "insert" as stated in page 16 section 30a and 30b of attachment 3.

300. EC062 – Attachment 14: The Southern blot analyses do not provide definitive answers about
the total number of copies of transgenic seque nces introduced into GS40/90pHoe6/Ac. Further
analyses (attachment 113, 114) will show that an additi onal copy to the two identified here are in fact
inserted into the nucleus of GS40/90pHoe6/Ac.
301. EC062 – Attachment 74: This Expert disagre es with the comments in paragraph 6.1.3
"Transgenic construct in the GM plant". So far, th e data presented only show that two functional loci
are present in GS40/90pHoe6/Ac. No structural analysis of progeny has been conducted to
demonstrate that only two transgenic loci were present and that each locus was structurally stable. The
Southern blot analysis presented in attachment 14 (see above) does not provide information about the
structure of each of the two transgenic loci. The use of the term "insert" is here again confusing as the
sentence "It is shown that the insert is stable a nd follows standard Mendelian inheritance" means in
fact that two functional loci have been identified and that the segregation of transgene phenotype (i.e.
herbicide tolerance) across generations is compatible with two functional Mendelian loci. This is not
completely informative about the structural stability or the number of transgenic loci (with the
exception that there is at least two of them).
302. EC062 – Attachment 113: Page 70 of the PDF file and onwards, what is "transgen" In
figures 1,2,3,5,6,7? Does "transgen" mean line GS40/90pHoe6/Ac?
303. EC062 – Attachment 113: This attachment contains, among many other documents, the study
A59211 entitled "Southern analysis performed on the gl ufosinate tolerant 6Ac rape line". This Expert
disagrees with the conclusion of this study that "no region outside the two border sequences of the
binary vector pHoe6/Ac are integrated into the geno me of the 6Ac rape line" stated page 69 of the
PDF file. The Southern blot analyses undertaken in this study seem to have been conducted on a rape
line containing only one copy of the CaMV35S:pa t:35T expression unit integrated into the plant
genome (the XbaI/XhoI digest hybridized with th e pat gene produces only one band in figure 5). This
is not compatible with the fact that GS40/90pHoe6/ Ac contains at least two functional loci (i.e. at
least two copies of the expression unit). Further molecular analyses should be conducted using
restriction enzymes which cut only once into the T-DNA and using the same set of probes 1,2,3,4 and
pat.
304. EC062 – Attachment 114: Page 5 should read "three copies of the T-DNA are inserted in
oilseed rape event falcon GS40/90 at two indepe ndent functional loci. It was shown that the
expression of the insert is stable and follows Mende lian inheritance". In the absence of molecular
analysis of the progeny plants, the exact number and stability of transgenic loci cannot be established.
305. EC062 – Attachment 114: The full sequence of the transgenic locus EE1 is provided with
flanking genomic sequences providing a full and optimal characterisation of this locus.
306. EC062 – Attachment 114: The sequence of the transgenic locus EE2 should be provided,
especially as it contains a combination of two T-DNAs . This reviewer could not locate the southern
blot analyses underlying the architecture proposed for EE2.
307. Conclusion – Response to Question 12: Since the first submission in 1996, the following
molecular information about GS40/90pHoe6/Ac has been established:
• GS40/90pHoe6/Ac contains two independent functional loci, named EE1 and EE2;
• The locus EE1 and its plant genomic flanki ng regions have been fully sequenced and
characterised. One copy of the CaMV35S:pat:35T expression unit is present in EE1;

• The locus EE2: A structural organisation involving two CaMV35S:pat:35T
expression units in tail-to-tail orientation has been proposed and plant genomic
flanking regions have been sequenced.
308. For a full molecular characterization to satisfy all regulatory requirements, this Expert would
suggest that more information is still required about:
• The locus EE2: the Southern blot data supporting the structural organisation of this
locus could not be found by this reviewer. It would be desirable to sequence the EE2
locus in order to provide the same type of complete information already available for
the EE1 locus.
• The molecular analysis of GS40/90pHoe6/ Ac for the presence of unwanted backbone
sequences needs to be re-visited as inconsiste ncies appear in the current analysis (see
attachment 113 – the plant analysed seems to contain only one pat transgene copy
and not three as expected from GS40/90pHoe6/Ac).
309. The structural analysis of progeny plants is required to ensure that no additional transgenic
sequences have been integrated in other genomic locations and that EE1 and EE2 are structurally
stable across generations.
310. Additional information may have submitted to the EC by the company. Is so, it should be
examined to see if it answers some of the requests formulated above.
Question 13: Given the information before the Pa nel, including the notification by AgrEvo and
the conclusions of the EC SCP in relation to the potential persistence or invasiveness of Bayer
oilseed rape (Falcon GS40/90), would this produc t qualify as a potential "pest" as the term is
used in ISPM 11?
Dr. Nutti
311. According to the International Plant Conve ntion's (IPPC) International Standard for
Phytosanitary Measures (ISPM 11) in EC-130, the defin ition of pest is "Any species, strain or biotype
of plant, animal or pathogenic agent injurious to plant or plant products". In my opinion, Bayer
oilseed rape (Falcon GS40/90) cannot be qualified as a potential "pest", according to this definition.
Dr. Andow
312. This question is a special case of the general c ase covered in my answer to question 6. My
answer to question 6 holds in this special case. Sp ecifically, it appears that on the issue of persistence
that the SCP did not explicitly consider the issue of release rate or scale, nor did it determine whether
contamination of conventional varieties was an inju ry to plants. Indeed, the data presented by the
notifier, the argument of the notifier, and the argum ent implicit in the SCP Opinion all suggest that
scale was not considered. Hence the SCP Opinion must be considered incomplete for allowing a
determination of "pest" status. I conclude that the pest status of Falcon GS 40/90 cannot be
determined based on the information before the Panel.
Dr. Snow
313. I conclude that in some situations, this product qualifies as a potential "pest", as I discuss in
Answer 6. If the herbicide glufosinate becomes used more widely, as expected, volunteers and weedy

Brassica rapa that develop resistance could qualify as worse "pests". However, this problem should
be manageable over the short term by using other methods of weed control.
Dr. Squire
314. In that oilseed rape as a volunteer or feral plant is a pest as defined in ISPM 11, e.g. a weed
competing with a crop for resources, and having ot her potential influences, the Bayer oilseed rape
(Falcon GS40/90) has the potential to be a pest, si nce it would leave volunteer and feral descendents.
Whether it would be more or less a pest than othe r forms of oilseed rape depends on whether it would
be sprayed as a volunteer or feral with the herbicide glufosinate ammonium (see Responses 6b-c).
That would be context-specific, e.g. whether farm ers in the region or country used this specific
herbicide widely or repeatedly in fields.
Bayer hybrid oilseed rape (MS8/RF3)
C/BE/96/01 (EC chronology 63)
Question 14: Given the information before the Pa nel, including the conclusions of the EC SCP
(EC-63/At.54) in relation to the potential persi stence or invasiveness of Bayer hybrid oilseed
rape (MS8/RF3), would this product qualify as a potential pest according to ISPM 11? If so, are
the proposed post-market monitoring plans consistent with the monitoring and review
principles described in ISPM 11?
Dr. Nutti
315. According to the International Plant Conve ntion's (IPPC) International Standard for
Phytosanitary Measures (ISPM 11) at EC-130, the definition of pest is "Any species, strain or biotype
of plant, animal or pathogenic agent injurious to plant or plant products". In my opinion, Bayer
oilseed rape (MS8 / RF3) cannot be qualified as a potential "pest", according to this definition.
Dr. Andow
316. Regarding the definition of potential pest, this question is answered more generally in my
response to question 6. That answer applies to this particular case. Specifically, it appears that on the
issue of persistence that the SCP did not explicitly consider the issue of release rate or scale, nor did it
determine whether contamination of conventional varieties was an injury to plants. Indeed, the data
presented by the notifier, the argument of the notif ier, and the argument implicit in the SCP Opinion
all suggest that scale was not considered. Hence th e SCP Opinion must be considered incomplete for
allowing a determination of "pest" status. I conclu de that the pest status of MS8/RF3 cannot be
determined based on the information before the Panel.
317. The need for risk management and monitoring was outlined in the SCP (EC-63/At.54, 19
May 1998) as follows:
2. The Committee was also of the opinion th at the potential transfer of the herbicide
resistance gene to wild Brassica relatives is a new issue in Europe in view of the
limited scale of release to date. The Comm ittee has examined the available evidence
from monitoring and research programmes to date. After evaluating all the
information available to the Committee, it was concluded that herbicide-tolerant
volunteers that may appear would be canola plants and not wild Brassica relatives.
Such herbicide-tolerant volunteers could be controlled in subsequent crops by

conventional agricultural methods. The Committee recommends that the introduction
of herbicide-tolerant crops should be accompanied by:
i) an agreed code of practice for the particular modified crop involving the active
participation of the notifier to promote best practice by farmers.
ii) a monitoring programme with an agreed design and implementation plan to detect
the occurrence and the establishment of herbicide-tolerant volunteers and weeds
under field conditions in the EU.
318. The SCP is recommending a "code of practice" in order to ensure that herbicide-tolerant
volunteers are eliminated. In other words, the purpose of the risk management measure is to eliminate
volunteers as they occur. In addition, it is re commending research design and implement monitoring
to detect herbicide-tolerant volunteers and herbicide-tolerant weeds in the EU.
319. In addition, the objectives of Annex VII, Directive 2001/18/EC also are relevant and are
acknowledged as such by the notifier (EC-63/At.124, p. 6).
320. The notifier proposes a two-part monitoring system: case-specific monitoring for HT
volunteers and HT weeds, and general surveillance fo r unanticipated effects (EC-63/At.124, p. 7ff).
These parts are intended to meet the concerns expressed by the SCP and the objectives of Annex VII.
321. Section 3.6.1 of ISPM 11 describes the monitoring and review of phytosanitary measures.
This concentrates on the principle of "modification", and is repeated here.
3.6.1 Monitoring and review of phytosanitary measures
The principle of "modification" states: "As conditions change, and as new facts
become available, phytosanitary measures shall be modified promptly, either by
inclusion of prohibitions, restrictions or requirements necessary for their success, or
by removal of those found to be unnecessary" (ISPM N° 1: Principles of plant
quarantine as related to international trade).
Thus, the implementation of particular phytosanitary measures should not be
considered to be permanent. After app lication, the success of the measures in
achieving their aim should be determined by monitoring during use. This is often
achieved by inspection of the commodity on arrival, noting any interceptions or any
entries of the pest to the PRA area. The information supporting the pest risk analysis
should be periodically reviewed to ensure that any new information that becomes
available does not invalidate the decision taken.
322. This suggests that an important part of th e risk management measures applied to GMHT
crops should include a process of review of the ne cessity of the management measures. Additional
general issues are discussed in my response to question 6.
323. The notifier has proposed a review process in the monitoring plan (EC-63/At.124, p. 21),
which is repeated here.

Review
This monitoring plan will be revised along with the expertise acquired. The
monitoring plan and its associated methodology will be reviewed at the end of each
growing season and updated as necessary in light of results obtained during the
monitoring programme. Reviews will examine the effectiveness and efficiency of
data measurements and collection in order to be able to improve the quality of the
monitoring programme.
324. In addition, the notifier proposes to terminate case-specific monitoring after five years.
325. Thus, I conclude that the proposed post-marke t monitoring plans are consistent with the
monitoring and review principles described in ISPM 11. The proposed plans are the most thorough
and well-considered of any of the monitoring plans in the material before the Panel.
326. However, I also believe that the proposed plan could be much improved so that it is not
merely consistent with ISPM 11 but actually meets the full intent of ISPM 11.
Dr. Snow
327. I conclude that this product could qualify as a potential pest, as I discuss in Answers 6 and 13.
If the herbicide glufosinate becomes widely used, as expected, glufosinate-resistant volunteers and
weedy Brassica rapa could become worse pests. Ho wever, this problem can be managed using other
methods of weed control. (note: the EC SCP conclusions about risks dealing with gene flow,
volunteers, and wild relatives are the same as tho se for Bayer oilseed rape, above, with the same
shortcomings; see Answer 11).
328. The ISPM 11 monitoring and review principles (section 3.6.1) discuss the need to modify
phytosanitary measures as needed when new information becomes available. The post-marketing
monitoring plans that are proposed by the notifi er are consistent with these principles. If
implemented, these plans would be useful for detecting the spread of glufosinate resistance to
volunteer plants and wild relatives.
329. Caveat – The monitoring plans do not address broader concerns about monitoring for changes
in biodiversity in and around farmland as a result of more efficient and extensive weed control, and
they do not provide unique identifying markers for monitoring gene flow (e.g., these issues are
discussed in the May 2004 Danish position; EC-063 At. 173).
Dr. Squire
330. The basic answer is similar to that given for Q.13. A few statements need comment,
however. The EC SCP (EC-63/At.54, section 6.3.1) gave the opinion that "rape is a poor competitor"
and "not regarded as an environmentally-hazardous colonising species", and while the latter is still
probably acceptable in semi-natural ha bitats, the continued widespread persistence of oilseed rape in
and around arable fields, both from original seed and from re-seeding, points to its competitive ability
being greater than "poor" but similar to many second -order weeds (i.e. weeds that are less important
than the most aggressive few species but are still important). The proposed post-marketing
monitoring plans (EC-63 At.060) are generally consistent with the principles in ISPM 11, but do not
give specific detail on, say, how a field is to be sampled to estimate % GM presence among
volunteers with a stated degree of certainty (see Responses 6e and 9).

Question 15: Given the information before the Pane l, including the conclusions of the EC SCP,
was the information regarding the assessment of the long-term effect of the newly expressed
protein on the biogeochemical cycle and the food chain requested by the Italian CA
(EC-63/At.87) necessary to ensure that conclusions of the safety assessment were valid?
Dr. Nutti
331. Based on the information provided by the appli cant (EC-63/At.01 to 176) and the analysis
done by the EC SCP presented in CDA-35A and EC-063/At.54, the information presented in CDA-56
and following the Codex Guidelines, my understandi ng is that the request by the Italian CA (EC-
63/At.87) was not necessary to ensure the conclusions for the safety assessment of the newly
expressed protein in the food chain. As pointed out by the EC SCP in item 6.2 – Safety Aspects, there
are no antibiotic resistance marker genes in this GM O. The bargene is controlled by a plant promoter
which is not functional in bacteria. Conseque ntly, its expression in the unlikely event of
transformation would not occur. Even if, due to ge netic recombination, the gene would be expressed
in intestinal micro-organisms or human or animal cells, the probability of which is remote, no
negative effects are expected b ecause the only known substrate of the enzyme phosphinothricin
acetyltransferase PAT is the herbicide glufosinate ammonium.
332. The PAT protein has been detected in very low amounts in dry seed (0.1 mcg/mg seed
protein). As almost no protein is present in the oi l extracted from the plants, the risk for human
consumption is non existent. The amounts of PAT (t he new protein) present in the seed-meal fed to
animals would be too low to cause even theoretical concern. The low amount of PAT protein in
vegetative tissues of the plants is an additional gua rantee of safety in case of occasional consumption
of the green parts of the plant by farm or wild an imals. The low levels of PAT protein and the weight
of evidence available elsewhere c oncerning the safety of PAT leads the Committee to conclude that
there is no significant risk to humans or livestock following the ingestion of the gene product.
333. Compositional analysis were carried out on the seed harvested from the original plant lines,
their hybrids, and lines developed from them by mu ltiple backcrosses to different oil seed rape
varieties from field trials in Europe and North America, where locally adapted commercial varieties
were used as controls. Information on fat content, gl ucosinolate levels, fatty acids profiles (including
erucic acid), vitamin E and mineral content showed no differences which could be attributed to the
genetic modification, leading to the assumption that no significant changes in nutritional value have
been identified.
334. I agree with item 7 of the Overall Assessment, where the EC SCP states that there is no
evidence to indicate that the placing on the market of hybrid seed of sweed rape (consisting of
crossing of parentals derived from genetically m odified sweed rape lines MS8 and RF3) with the
purpose to be used as any other sweed rape is lik ely to cause any adverse effects on human health.
Based on the scientific evidences presented, my understanding is that the new protein is safe for
consumption in the food chain.
Dr. Andow
335. In EC-63/At.87, 14 March 2000, the Italian CA requested the following information.
Referring to the conclusion of the last meeting of the Regulatory Committee under
Directive 90/220/EEC, Italy would be very pleased to receive from the notifier more
information concerning the assessment of the effect of the genic product on the

biogeochemical cycles and on food chain and on the apreading of the gene due to the
possibility of crossing between the PGM and wild species.
336. This is an identical request for information on the same date as for Falcon GS 40/90 (EC-
62/At.95). Even without looking into the specifics of this case, my genera l comments on this request
to the notifier of Falcon GS 40/90 in my response to question 10 in paragraph 252 also holds in this
case.
337. The Italian CA made the request on 14 Ma rch 2000. The notifier provided an updated
notification on 10 November 1999, which was circulated to the CAs on 13 December 1999. The
notifier provided additional information on 25 February 2000 and 1 March 2000. Except for the dates
that additional information was provided, this is the same timeline as Falcon GS 40/90, and my
conclusion in paragraph 256 holds here.
Question 16: Given the information before the Panel, including the conclusions of the SCP, was
the information regarding molecular characterizati on of this product requested by the lead CA
(EC-63/At.107) necessary to ensure that conclusions of the safety assessment were valid?
Dr. Healy
338. Following receipt in September 1996 of the notification from Plant Genetic Systems (PGS,
now Bayer Crop Science) seeking approval for oilseed rape lines MS8 and RF3, the lead CA
(Belgium) expressed a favourable opinion in January 1997. The dossier was then sent to the European
Commission and distributed to other Member States for comment. Supplementary information from
PGS required by Member States was added to the dossier, which was submitted to the SCP for
assessment. In May 1998, the SCP also expressed a favourable opinion on MS8, RF3 and hybrid
MS8xRF3 lines with respect to human health and safety.
339. Background: The two oilseed rape lines under c onsideration, MS8 and RF3, both contain a
bacterial gene, pat, that confers tolerance to the herbicide phosphinothricin, also known as
glufosinate-ammonium. The presence of the pat gene enables the rape plants (Brassica napus) to
produce an enzyme, phosphinothricin acetyl transferase (PAT), which chemically inactivates the
herbicide, allowing the plants expressing the PAT protein to function normally in the presence of the
herbicide. The pat gene was used as a selectable mark er gene in the individual lines and also to confer
tolerance to commercial applications of glufosinat e-ammonium herbicide in MS8/RF3 hybrids. There
were no other marker genes used and there are no antibiotic resistance genes in either MS8 or RF3.
340. In conjunction with the herbicide tolerance trait, MS8 contains another bacterial gene,
barnase. Expression of barnase in specific parts of the flower at a particular developmental stage gives
rise to plants that do not produce pollen (male-st erile, MS). The barnase gene prevents pollen
formation by producing a non-specific ribonuclease that destroys the tapetal cell layer in which it is
expressed.
341. The RF3 line contains the corresponding bacteria l gene, barstar, as well as the herbicide
tolerance trait. Expression of the barstar gene in the same floral tissues, the tapetal cells, produces a
protein that specifically binds and inactivates the barnase, thereby restoring normal pollen production.
Hybrid MS8/RF3 plants are phenotypically normal and fully fertile. The primary objective of the
modifications in MS8 and RF3 is to generate pollin ation-controlled parental oilseed rape lines with
superior agronomic performance for use in a breeding system that generates hybrids with significantly
higher seed yields.

342. Molecular characterisation data: PGS submitted a range of company studies on the molecular
characterisation of MS8 and RF3 lines in its origin al dossier. These data were consistent with the
approach generally considered appropriate at the ti me for the assessment of the safety of transgenic
crop plants and derived food products intended for human consumption.
343. Descriptions of the plasmids pTHW107 and pTHW118 used to generate the MS8 and RF3
lines respectively were provided, including the size, source and function of the transgenes and their
regulatory elements. In addition to standard Southern blot hybri disation analyses that generate
information on the number and size of insertions, the data included DNA primer sequences, PCR
methods for analysing the specific insertion ev ents, and detailed DNA sequence analyses of the
integration region in both lines. The CA (Belgium) declared in 1997 that the data provided showed
that the transgenes in lines MS8 and RF3 are not inserted into a known functional plant gene (EC 063,
attachment 007).
344. Approximately 4900 base pairs (bp) of the inserted DNA sequence in MS8 was provided,
showing exact similarity with the transforming plasmid sequence between the left and right borders of
the T-DNA, with no deletions or rearrangements. Flanking nucleotide sequence was provided at the
5'end (864 bp) and the 3'end (357 bp) of the inser ted DNA. These flanking sequences were analysed
for homology with known sequences. Part of the 5' flanking sequence showed some degree of
similarity with published sequences from Arabidops is thaliana chromosomes 5 and 3. However, no
homology with known sequences was found using the 3' flanking sequence.
345. The DNA sequencing analyses of RF3 showed that the inserted DNA corresponds exactly
with the transforming plasmid, however from the data it was evident that an inversely-oriented second
partial fragment of T-DNA was present at the left bo rder of the introduced DNA at the same insertion
site. The second copy includes a functional part of the anther specific TA-29 promoter, the coding
region of the barstar gene, 3' nos termination sequence and a non-func tional part of the PssuAra with
the T-DNA sequence ending 750 bp upstream of the ATG initiation codon of the bar gene. Detailed
nucleotide sequencing across the integration sites generated 1077 bp at the right border, 1457 bp at the
inverted repeat junction, and 1501 bp at the left border. Approximately 8900 bp corresponding to the
entire RF3 insertion event was determined. Flanking nucleotide sequence was provided at the 5' end
(812 bp) and at the 3' end (1275 bp) of the inser ted DNA. These flanking sequences were analysed for
homology with known sequences. Parts of the 3' flan king sequence showed some degree of similarity
with published sequences from Arabidopsis thaliana chromosomes 1 and 5. However, no significant
homology with known sequences was found using the 5' flanking sequence.
346. Conclusions of the Safety Assessment: The SC P considered in 1998 that the data submitted
in the dossier was satisfactory concerning information on the transformation techniques, vector
constructs, insertion event in the transgenic plan ts and transgene and cryptic gene expression. The
overall assessment and conclusion of the SCP was that 'there is no evidence to indicate that the
placing on the market of hybrid seed of swede rape (consisting of crossings of parentals derived from
genetically modified swede rape lines MS8 and RF3) with the purpose to be used as any other swede
rape is likely to cause adverse effects on human health and the environment'.
347. Additional Information Sought by Lead CA: Although junction regions at the sites of
integration in the transformed lines were separately cloned and sequenced, there was minimal DNA
sequence data generated from the wildtype isogenic lines at the corresponding locus. Consequently,
the submitted homology searches were performed using sequence data obtained only from the
transgenic lines. Subsequently, molecular details of the flanking regions were not considered to be
adequate by the lead CA and the results of further PCR and sequence analyses were sought.

348. Impact of Additional Information on the C onclusions of the Safety Assessment: The
additional molecular characterisation data sought by the lead CA in 2002 were not required to
demonstrate the safety of food products derived from MS8, RF3 or MS8xRF3 hybrid lines. The
rational for this view are:
• Finely detailed molecular analysis of a defined chromosomal region in commercially
adapted plant lines such as oilseed rape is likely to reveal minor nucleotide
differences between various lines that have no significance with respect to the safety
of derived food products. In the absence of an equivalent nucleotide analysis from a
number of commercial food-producing oilseed rape lines, these data do not contribute
substantially to the bank of information th at is necessary to adequately characterise
MS8 and RF3 at the molecular level. Moreover, it is not clear how such data should
be interpreted in an assessment of transgenic plants.
• The data provided at that point in time (i) allowed adequate description of the
insertion events in both transgenic lines, (ii) was sufficiently detailed, presented
clearly and of acceptable quality, and (iii) was consistent with a generally accepted
approach to the molecular characterisation of transgenic crops used for food
production.
• The most significant elements of the assessment of MS8 and RF3 oilseed rape lines
more appropriately focus on the molecular characterisation of the insertion events and
the resultant novel gene products, particularly in terms of their potential toxicity and
allergenicity. The food products derived from transgenic oilseed rape are likely to
contain at most only trace amounts of plant proteins due to processing. Comparative
compositional data from the seeds of bot h transgenic lines and the corresponding
isogenic lines therefore provides necessary information to determine unintended
effects.
349. Conclusion: The totality of information provided in the dossier to that point in time contained
sufficient information to support the validity of th e conclusions of the safety assessment without the
need for the additional molecular ch aracterisation data requested in the letter of the Biosafety Council
dated 28 March 2002.
Dr. Snape
350. The comments below relate to Questions 16 and 18 concerning the molecular characterisation
of exhibit EC-063. These were examined relative to the EFSA-Q-2003-005 guidance document
produced by the Scientific Panel on Genetically Modified Organisms.
351. Oilseed rape event Ms8 was produced, using plasmid pTHW107, by means of
Agrobacterium-mediated gene transfer. The T-DNA of the plasmid pTHW107 used for
transformation carries two expression units: one contai ns the bar gene, conferring resistance to the
herbicidial compound glufosinate and one contains the barnase gene to engineer male sterility. A
barstar gene (driven by a bacterial promoter) is also present in the backbone of pTHW107. Oilseed
rape event Ms8 is described as containing one functi onal transgenic locus containing one copy of the
pTHW107 T-DNA.
352. Oilseed rape event Rf3 was produced, using plasmid pTHW118, by means of Agrobacterium-
mediated gene transfer. The T-DNA of the plasmid pTHW118 used for transformation carries two
expression units: one contains the bar gene c onferring resistance to the herbicidial compound
glufosinate and one contains the barstar gene to engineer fertility restoration. An additional copy of
the barstar gene (driven by a bacterial promoter) is also present in the backbone of pTHW118. Oilseed

rape event Rf3 is described as containing one functio nal transgenic locus containing two copies of the
pTHW118 T-DNA in inverted orientations, one copy of the T-DNA being intact the other one being
partially deleted.
353. Information relative to the molecular charact erisation of Ms8 and Rf3 has been found
primarily in EC-063 attachments 4, 19, 20, 43, 54, 98, 114, and 115. A series of comments and
questions are numbered below. This is followed by a conclusion proposing answers to Questions 16
and 18:
354. EC063 – attachment 001: (1) Contains only the table of content of the initial notification. The
data corresponding to the molecular analysis in part1 section III and IV (pages 7 to 20) were not
found by this Expert in subsequent attachments. When requested, this info rmation was not available
from WTO either. It is likely that some relevant information was initially submitted by the company.
However in its absence this Expert cannot determine:
(1a) The sequence of pTHW107 and pTHW118
(1b) If oilseed rape event Ms8 contains only one copy of the bar gene and one copy of the
barnase gene.
(1c) If oilseed rape event Rf3 contains only one copy of the bar gene and two copies of the
barstar gene.
355. EC063 – attachment 019 and 020: Presents a PCR analysis suggesting the absence of Sm/Sp
antibiotic resistance gene in Ms8 and Rf3.
356. EC063 – attachment 043: Presents a Southern blot analysis demonstrating the absence of
vector backbone sequences into Ms8 and Rf3 including the barstar gene in Ms8.
357. EC063 – attachment 098: The transgenic locus in Ms8 and its plant genomic flanking regions
have been fully sequenced and analysed providing an optimal characterisation of this locus. This
transgenic locus contains one copy of the pTHW107 T-DNA.
358. The transgenic locus in Rf3 and its plant genomic flanking regions have been fully sequenced
and analysed providing an optimal characterisation of this locus. This transgenic locus contains two
copies of pTHW118 T-DNA in inverted orientations , one copy of the T-DNA being intact the other
one being partially deleted.
359. Conclusion – Response to Questions 16 and 18: Additional information may have been
submitted to the EC by the company (see point 1a, 1b, 1c above). Experts should examine if this
information answers some of the requests formulated below.
360. Ms8: Since the first submission in 1996, the following molecular information about Ms8 has
been established:
• Ms8 contains one transgenic locus (if confirmed by Southern analysis see point 1b).
• This locus and its plant genomic flanking regions have been fully sequenced and
characterised. One copy of the pTHW107 T-DNA is present at this locus.
• Transgenic sequences outside the T-DNA of pT HW107 have not been integrated into
Ms8.

361. This Expert concludes that no more informati on is required (if Southern analysis confirm
point 1b). As Ms8 contains a single copy of the bar and barnase gene, the monitoring of transgene
expression across generations provides a satisfactory estimation of the transgenic locus structural
stability and inheritance.
362. Rf3: Since the first submission in 1996, the following molecular information about Rf3 has
been established:
• Rf3 contains one transgenic locus (If confirmed by Southern analysis see point 1c).
• This locus and its plant genomic flanking regions have been fully sequenced and
characterised. Two copies of the pTHW118 T-DNA in inverted orientations are
present at this locus, one copy of the T-DNA being intact the other one being partially
deleted.
• Transgenic sequences outside the T-DNA of pTHW118 to have not been integrated
into Ms8.
363. In the opinion of this Expert, to satisfy all re gulatory requirements, more information is still
required about the structural analysis of progeny plants to ensure that the transgenic locus is
structurally stable across generations. As Ms8 cont ains two copies of the barstar gene the monitoring
of transgene expression across generation is not in formative on the stability and inheritance of the
transgenic locus.
Question 17: Were detection meth ods commercially available in 2001 sufficient to enable the
detection of the transgenic proteins expressed by the plant line hybrid oilseed rape MS8/RF3?
Given the information before the Panel, including the SNIF (EC-63/At.109) and the updated
environmental risk assessment (EC-63/At.110-14 0), was additional information regarding a
quantitative detection method (EC-63/At.141) necessa ry to ensure that conclusions of the safety
assessment were valid?
Dr. Nutti
364. A qualitative method available at the time. EC-063/At.141 asks for a quantitative event
specific detection tool for elite events and hybrids. It is recognized that a qualitative detection method
and reference materials were provided by the co mpany. Based on the information provided at CDA-
56, I can infer that there was no general validation of the method used by the notifier. So far, only
DNA extracted from unprocessed raw mate rial has been tested. It was observed that this detection
method should also be demonstrated to work usi ng other matrices. Further, the sensitivity of the
detection method should still be assayed by testing th e presence of the elite or hybrid events in mix
samples. Finally, based on the given method, one can not make a distinction between the hybrid and a
mixture of the two parental lines. My conclusi on is that the method presented was technically
sufficient for the safety assessment, although it would have to be validated.
365. Although a quantitative method will provide rele vant information for labelling and to the
consumer, it is not necessary to ensure the validity of the safety assessment. It is important to point
out that the EC SCP has considered that there is no evidence to indicate that the placing on the market
of hybrid seed of oilseed rape (c onsisting of crossing of parentals derived from genetically modified
oilseed rape lines MS8 and RF3) with the purpose to be used as any other oilseed rape is likely to
cause adverse effects on human health.

Dr. Healy
Question 17 – part 1:

366. The transgenic oilseed rape lines MS8 and RF3 are developed specifically to generate high
yielding seed produced by the MS8xRF3 hybrid. Fo od products intended for human consumption are
therefore derived from the hybrid seed rather than from the individual parental lines. Novel transgenic
proteins expressed in the hybrid plants include th e PAT protein (herbicide tolerance), and the two
bacterial proteins barnase and barstar (for pollinatio n control in the parental lines). The patterns and
levels of gene expression conformed to those predicted and intended by the modification process.
367. The SCP acknowledged that the dossier containe d appropriate information on PCR detection
methods for identification of MS8 and RF3 transgenic lines (Opinion, 19 May 1998).
368. Barnase and barstar: The barnase and barsta r genes, derived from the bacterium Bacillus
amyloliquefaciens, each encode a different small si ngle-chain protein. Both of these proteins have
been studied extensively as models for protein fold ing because of their small size, and there is an
abundance of published scientific information relati ng to research work conducted since the early
1960s.
369. The barnase gene encodes a ribonuclease that is naturally secreted by the bacterium.
Ribonucleases are ubiquitous in nature, and serve many biological functions. In this case, the secreted
ribonuclease serves to protect the environment of the bacteria. Conversely, the barstar gene encodes a
specific protein inhibitor of this ribonuclease. In th e Bacillus species from which the two proteins are
derived, the function of the barstar protein is to protect the organism from the otherwise toxic effects
of its own barnase activity. This naturally occurring system is well studied and the interaction of the
two proteins is known to be highly specific.
370. Under the control of a highly tissue-specific plant promoter from tobacco (PTA29),
expression of barnase and barstar in the MS8xRF3 hy brids is restricted exclusively to the pollen
producing cells in developing anthers and only when the plants are flowering. The two proteins are
not expressed elsewhere in the plant, including the leaves, stems or seeds.
371. Gene expression data: For the parental RF3 line, barstar messenger RNA (mRNA) was
barely detected in flower buds only, but not in any other plant tissues, including the seeds. As
expected, because expression of the ribonuclease resu lts in cell death, barnase mRNA could not be
detected in any tissues from the MS8 line.
372. Expression of PAT protein: The enzyme responsible for herbicide tolerance, PAT, is mainly
expressed in the green tissues of th e plant. There is some expres sion in other tissues including the
seeds, but at such low levels that specific en zyme activity was not detectable. Messenger RNA
(mRNA) corresponding to the bar gene could be detected at extremely low levels in the leaves and
flower buds of parental lines MS8 and RF3, but not in the seeds of the plants.
373. Hybrid MS8xRF3 would be expected to express hi gher levels of PAT than either parental
line. Data were provided indicating that the levels of PAT expressed in the seeds of the individual
parental lines were detectable using the speci fic Enzyme Linked Immunosorbent Assay (ELISA)
system, with a detection limit of approximately 5 ng/ml. As expected, the level of PAT protein in
various oil fractions of seeds derived from the MS8xRF3 hybrid is below the limit of detection.

374. Protein detection methodology: As well as detailed DNA-based detec tion protocols, the
company provided protein-based detection and identification methodologies (ELISA and test kit)
which were evaluated at the September 2001 meeting of the Scientific Committee "Transgenic Plants"
of the Biosafety Council (EC 63-30). In its discuss ions of the information, the Committee raised
several matters concerning the protocol for detec tion of the PAT protein using ELISA. Concerning
amounts of PAT protein detected, there were questions raised on the use of the standard curve and on
the limit of detection. It was also remarked that the detection methods proposed by the company were
qualitative rather than quantitative.
375. However, quantitative immunoassay methods to detect novel proteins in foods derived from
transgenic crops were under development during th e late 1990's. The two most common tests are
ELISA and lateral flow strips. Commercial immu noassay methods were reportedly available for
detection and quantification of a number of transg enic proteins including the PAT protein in 2000 59.
As there are numerous transgenic food crops that ha ve been genetically modified for tolerance to the
glufosinate-ammonium herbicide, detection of PAT in the MS8xRF3 hybrids is not an event-specific
detection method. Information supplied by Strategi c Diagnostics Inc (Delaware, USA) indicates that
their Lateral Flow test for oilseed rape was commercially available in April 2002.
376. Conclusion: The available evidence indicat es that immunoassay-based detection methods
suitable for the detection of PAT protein in MS8xRF3 hybrid transgenic plants were commercially
available as early as 2000.
Question 17 – part 2:

377. Additional information regarding the provision of an event-specific quantitative detection tool
was sought on 25 February 2003 (EC 63-141). The re quest for a detection method that is "event-
specific" directly implies the use of a DNA-based method rather than a protein-based method, which
can be quantitative but in this case is not event-specific.
378. The company provided both DNA-based (PCR primers and protocols) and protein-based
methods (ELISA and test kit) for detection and iden tification of its transgenic lines. This information
together with reference material was used in an independent laboratory to evaluate the detection
protocol. The results of this evaluation indicated that the DNA-based PCR detection methods were
reproducible and valid for the transgenic oilseed rape lines MS8 and RF3.
379. The provision of event-specific oligonucleotide primers for unambiguous identification of
transgenic plant lines readily enables use of qualitative PCR methods, but also allows development of
quantitative PCR technology according to plant species. Quantitative PCR methods have been
developed for a range of food commodities including soybean and maize products but validation of
these methods is not straightforward. Moreover, it is generally recognised that quantitative protein-
based methods of detection are more directly accessible.
380. Quantitative detection is an integral part of enforcement capabilities especially concerning
labelling regulations. However, it is not a necessary component of a safety assessment. Given the
comprehensive nature of the information and ma terial provided in the updated dossier regarding
identification primers, protocols and plant-deri ved DNA reference material, and the existence of

59 International Life Sciences Institute (ILSI) Euro pe Novel Food Task Force in collaboration with the
European Commission's Joint Research Centre (JRC) and ILSI International Food Biotechnology Committee,
Joint Workshop: Method Development in Relation to Re gulatory Requirements for the Detection of GMOs in
the Food Chain, December 2000. Journal of AOAC International, Vol. 85, No. 3, 2002.

established quantitative protein-based detection methods , I consider that further information relating
to quantitative event-specific detection tools for tr ansgenic oilseed rape lines MS8 and RF3 is not
necessary to ensure that conclusions of the safety assessment were valid.
381. Conclusion: Additional information regardi ng an event specific, quantitative detection
method requested in February 2003 is not necessary to ensure that the conclusions of the safety
assessment are valid.
Question 18: Given the information before th e Panel, including the SNIF and the updated
environmental risk assessment (referenced ab ove), was the information regarding molecular
data requested by the CA (EC-63/At.144) necessar y to ensure that conclusions of the safety
assessment were valid?
Dr. Healy
382. The original dossier was supplemented with a dditional molecular characterisation data and
other information to conform with particular requi rements of the Belgian Biosafety Council, the SCP
and Member States. The totality of the data a nd information provided allowed a comprehensive
assessment of the safety of food and feed derived from transgenic oilseed rape lines MS8, RF3 and
hybrids, as well as the potential environmental impact. Previously, both the Biosafety Council and the
SCP had declared that these lines do not pose human health and safety concerns, and the additional
information sought between 1998 and 2002, and sub sequently provided by the company, had not
altered these favourable conclusions.
383. Additional Information Sought by CA: The new Belgian guidelines on molecular
characterisation were developed over a period of time culminating in the final draft version
(version 6) being distributed for comments to th e Scientific Committee in November 2001. This
version was submitted to the members of the Biosaf ety Council for final approval at the meeting of
February 2002 (EC-63/At.105). Additional informat ion was sought 'in order to bring the molecular
data in the file into line with the new Belgian guidelines on molecular data: evaluation of the presence
and functionality of new chimeric ORFs via bi oinformatics on the sequence of the insert and the
flanking areas'.
384. It is noted that more detailed requirements c oncerning molecular characterisation data were
required by Belgium prior to September 2001. In r esponse, an extended package of information was
provided by the company, and this additional inform ation was evaluated at tw o subsequent meetings
of the Scientific Committee "Transgenic Plants " on 27 September 2001 and 7 February 2002. After
some modifications, the Scientific Committee approved the information supplied in these additional
documents. It is also noted however that the new Belgian guidelines were deve loped in parallel with
the ongoing assessment of the dossier and requests to the company for further molecular information.
It is surprising therefore that the informati on necessary to comply with the new molecular
characterisation guidelines was not requested until July 2003.
385. Impact of Additional Information on the Safe ty Assessment: Given the pace of advancing
knowledge in the field of molecular biology and plant biotechnology and the rapidly evolving
technical capabilities, for a foreseeab le time, there will often be a discrepancy between the scientific
data submitted in a dossier for assessment, and the information possible from the most recently
developed experimental techniques. In the specifi c case of the frequently updated dossier on MS8,
RF3 and hybrids, it is clear from the wording of the request to the company that the additional
information sought is for the purpose of "simply" ma king the file consistent with the current Belgian
guidelines on molecular data. This additional inform ation does not add substantially to the weight of

evidence that supports the lack of human health an d safety concerns associated with the use of the
transgenic oilseed rape.
386. In this constantly changing environment, the data requirements for the molecular
characterisation of transgenic crops are not static. In general, there is a recognised need to constantly
monitor the progress of technical advances and adapt regulatory guidelines accordingly as techniques
move from the highly experimental area to more validated and established methodology. It is
acknowledged for example that newly evolving research fields such as proteomics and metabolomics
may play a strategic role in the future assessment of food products derived from transgenic organisms.
387. Conclusion: The additional information sought by the CA in the email of 14 July 2003 (EC
63-144) on molecular data was requested to upda te the dossier to new guidelines and was not
necessary to ensure the conclusions of the safety assessment were valid.
Dr. Snape
388. See answer given to Question 16
Question 19: Given the information before the Panel, including the SNIF, the updated
environmental risk assessment and the clarific ation provided by the notifier (EC-63/At.147),
was the information regarding ecological effect s of this product on agricultural systems
requested by the lead CA (EC-63/At.149) necessary to ensure that conclusions of the safety
assessment were valid?
Dr. Andow
389. The lead CA requested the following information (EC-63/At.149, 10 October 2003): "The
Council found a shortcoming highlighted in the report to be sufficiently serious in terms of Annex 2,
D2 points 4, 5, 7, 8, 9 of Directive 2001/18: the data on file are not precise enough to inform the
Council properly about the impact of commercial crops of Ms8 or Rf3 or their hybrids, in the
presence of glufosinate on the parameters set in the aforementioned points in agricultural ecosystems.
Further data are awaited concerning: farmland dive rsity (macro-fauna, weed flora and microbial soil
ecosystem), food web integrity (trophic structur e), population dynamics of key species, life-cycles,
etc."
390. The lead CA is referring to Directive 2001/18/ EC Annex 2, section D2, which is reproduced
here:
D2. In the case of genetically modified higher plants (GMHP);
4. Potential immediate and/or delayed environmental impact resulting from direct and
indirect interactions between the GMHP and target organisms, such as predators,
parasitoids, and pathogens (if applicable);
5. Possible immediate and/or delayed environmental impact resulting from direct and
indirect interactions of the GMHP with non-target organisms, (also taking into
account organisms which interact with ta rget organisms), including impact on
population levels of competitors, herbivores, symbionts (where applicable), parasites
and pathogens.;
7. Possible immediate and/or delayed effects on animal health and consequences for
the feed/food chain resulting from consumption of the GMO and any products
derived from it if it is intended to be used as animal feed;

8. Possible immediate and/or delayed effects on biogeochemical processes resulting
from potential direct and indirect interacti ons of the GMO and target and non-target
organisms in the vicinity of the GMO release(s);
9. Possible immediate and/or delayed, direct and indirect environmental impacts of
the specific cultivation, management and harvesting techniques used for the GMHP
where these are different from those used for non-GMHPs.
391. The notifier's original response to these A nnex 2 guidelines is given in EC-63/At.112,
14 January 2003.
392. The notifier's full response to point 4 60 was "This is not applicable to glufosinate tolerant
oilseed rape." This is an insuffi cient response. At a minimum there should be an argument as to why
the point is not applicable. I conclude that the request by the lead CA for more information on point 4
was necessary to ensure that conclusions of the safety assessment were valid.
393. The notifier's response to point 5 covers 4 pages.61 The notifier concludes
– Scientific experts confirm that the hor izontal gene transfer from the GMHP to
bacteria is extremely unlikely although theoretically it cannot be totally
excluded. In the very unlikely case wher e such a transfer would occur this
would have no impact since the transgenes involved would not provide a
selective advantage.
– Scientific experts also confirm that the GM OSR has no effect on honeybees
and epigaeic predatory arthropods of oilseed rape.
– There are no identified potential effe ct of the GMHP on birds and small
mammals.
394. I believe that the notifier's first conclusion is well supported by the data and arguments they
muster. I believe that the data on epigeal predator s is weak, but even with additional data, I am not
convinced that a major adverse effect will be identi fied. The data on birds and rabbits are also weak,
but again, I am not convinced that additional data will reveal a major adverse effect.
395. As there are literally hundreds of species of non- target organisms that could be tested, it can
always be argued that more species should be evaluated. Indeed, several important ecological
functional groups were not evaluated, including parasitoids and detritivores. Without a specific
reason, it is difficult to argue that testing a paras itoid is necessary for the risk assessment. A case
could be made that a detritivore would have been a better choice for study than epigeal predators.
However, if this was the specific concern of the lead CA, it would have been appropriate to clarify the
point. Hence, I conclude that the request by the lead CA for more information on point 5 was not
necessary to ensure that conclusions of the safety assessment were valid.
396. The notifier's response to point 7 covers 3 pages. 62 The notifier argues that there is no reason
to believe that the feed value of GMHT oilseed ra pe will be different from conventional oilseed rape,
provides a summary of data on the major nutrients of seed and meal, and report the results on a rabbit
and chicken feeding study. It is possible that the compositional studies are not adequate or the wrong

60 EC-63/At.112, p. 28, 14 January 2003.
61 EC-63/At.112, pp. 29-32, 14 January 2003.
62 EC-63/At.112, pp. 37-39, 14 January 2003.

species have been used for feeding studies. I am unable to conclude that the request by the lead CA
for more information on point 7 was necessary or not necessary to ensure that conclusions of the
safety assessment were valid.
397. The notifier's response to point 8 is one page. 63 The notifier claims that oilseed rape does
participate in biogeochemical cycles, therefore there are no expected effects on biogeochemical
cycling. The notifier further supports this view by reporting that there have been no observed effects
on rhizoshere bacteria and no observed effects on the plants growing in the field the year following a
crop of GMHT oilseed rape.
398. The notifier is mistaken to claim that oilseed rape does not participate in any biogeochemical
cycles. The root and leaf litter participate in carbon cycles, and it may have subtle effects on
phosphorous cycles, as it will reduce proliferation of my corrhizal fungi relative to most other plants.
It is likely that participation in carbon cyclin g is one of the main ways it participates in
biogeochemical cycles. A simple litter degradation st udy would determine if it is likely to have any
effects on carbon cycling. Hence, I conclude that the request by the lead CA for more information on
point 8 was necessary to ensure that conclusions of the safety assessment were valid.
399. The notifier's response to point 9 covers two pages.64 The notifier argues
The only change in cultivation techniques used on the herbicide tolerant (hybrid)
oilseed rape is the possibility to use glusof inate ammonium as selective herbicide.
However, glusofinate-containing herbicide can be used on the crop only if they are
registered for this specific use in the different Member States according to Directive
91/414/EEC. The safety assessment and the assessment of the impact of the
herbicide use on the environment are done in the said 91/414 application.
400. The notifier argues that except for herbicide use, other cultivation practices are unlikely to
change. This contrasts sharply with claims by others that GMHT crops are likely to result in changes
in tillage practice. The notifier does not address th is possibility in detail a nd states only that soil
preparation practices will be identical. This seem s to be an insufficient argument. In addition,
although the UK-FSE trails will not be published until later in 2003, everyone involved was aware of
the experiments and the possibility that the associated herbicide use would have adverse
environmental effects. These issues are not addr essed in the response of the notifier. Hence, I
conclude that the request by the lead CA for more information on point 9 was necessary to ensure that
conclusions of the safety assessment were valid.
401. The lead CA requested information on 10 October 2003. The notifier submitted the material
to be reviewed on 14 January 2003. The 269 days th at elapsed seem excessively long to complete the
review and articulate the questions.
Dr. Snow
402. The vagueness of this request and the lack of related information from the notifier make it
difficult to provide a clear answer to this question from the Panel.
403. The lead CA asked for inform ation about effects on "farmland diversity (macro-fauna, weed
flora and microbial soil ecosystem), food web inte grity (trophic structure), population dynamics of
key species, life-cycles, etc." (EC-63/At. 149 translation). Although this request seems to ask for

63 EC-63/At.112, p. 40, 14 January 2003.
64 EC-63/At.112, pp. 41-42, 14 January 2003.

some unnecessary data (and the request is quite vague), several Member States were concerned about
the ecological effects of more intensive herbicide use. I assume that th is is the focus of concern – that
more information was requested to determine the e ffects of cultivating new herbicide-tolerant crops
on the weed plants that support insect and animal populations in farmland habitats in these countries.
404. Realistically, this is a difficult set of questions to answer, regardless of whether one uses basic
theory or empirical research. As noted above, the indirect effects of introducing herbicide-tolerant
crops to countries like the UK are still being investigat ed, so in this sense th e requested information
could be considered as necessary. However, it not feasible to obtain scientific data on all of the
processes listed by the CA without extensive multi-year studies. Even the Farm Scale Evaluations are
too short and too small-scale to answer these questions (e.g., Andow 2003, Squire et al. 2003).
Adverse effects on flora and fauna are mentioned in SPS and ISPM 11, but it is not possible to test for
all of these effects prior to deregulation. Inst ead, scientific committees typically use their best
judgement to recommend approving or denying applications for marketing.
Dr. Squire
405. The updated environmental risk assessment is comprehensive on topics for which data are
available. The further information on ecological ef fects requested by the lead CA was clearly not
available, and could only be in existence if there had been either large scale experimental
measurements on the habitats typical of where th e crop would be grown commercially or specially
commissioned experiments on trial crops, as in the UK's Farm Scale Evaluations. Since cropping with
this variety and its herbicide has potentially new effects on the arable flora (i.e. different from those of
other oilseed rape varieties with other herbicides) , which may be severely depleted in any case, it is
legitimate to ask what such effects might be. However, the general matter of the need for a
comparator is relevant here (Notes, paragraphs
32-35). Surely it is incumbent on both sides in the
argument to proffer their standard or comparator against which the new technology should be judged.
Question 20: Are the conclusions and recommendati ons in the Report of the Group of Experts
Mandated by the Belgian Biosafety Advisory Council (EC-63/At.167 pp.20-21) regarding the
agricultural guidelines proposed by the applicant consistent with the Belgian Biosafety Advisory
Council's conclusion that the agricultural guidelines are "impracticable, hardly workable and
hard to control"?
Dr. Andow
406. The Belgian Biosafety Council concluded (EC-63/At.166, 27 January 2004) as follows
Concerning the environemental risk, the Biosafety Advisory Council is of the opinion
that some risks associated with these GM crops will be reduced provided that the
following conditions are strictly applied:
– The agricultural guidelines must be (1) adapted to the recommendations of
the experts and to the recommendations made by the Member States due to
differences in agricultural practices a nd landscapes, (2) applied by all the
operators supported by appropriate legislation and control, and (3)
continuously updated according to the new available scientific information.
– The post-market monitoring plan must (1) assess compliance with the
agricultural guidelines, (2) assess and determine the effectiveness of the
proposed risk management strategies of the agricultural guidelines in limiting

gene flow and its potential conse quences and in preserving farmland
biodiversity, (3) be adapted to the recommendations made by the Member
States of the European Union due to di fferences in agricultural practices and
landscape, (4) be continuously upda ted according to the new available
scientific information and (5) the no tifier must provide a more detailed
description of the tasks and the kind of data that will be collected by different
networks involved in the general su rveillance. In this context the
detection/identification method must be validated.
Furthermore, the Biosafety Advisory Council considers that presently, a number of
recommendations of the agricultural guide lines are impracticable, hardly workable
and hard to control in current agricultura l practices, hence vertical gene flow and
negative consequences on biodiversity may not be controlled.
407. The expert recommendations referred to are (EC-63/At.167, 3 February 2004, pp. 20-21)
408. 4.2.3. Agricultural guidelines
Taking the importance of the agricultural guidelines into account, the Group of
Experts recommends that the agricultural guidelines should be translated in the
national or local language(s) of each MS of the EU. The guidelines should be
readable and understandable by all the opera tors (e.g. farmers, seed suppliers) that
will handle the seed, crop and product of MS8, RF3 and MS8xRF3. A widely
comprehensible language should be used. Moreover, the agricultural guidelines
should be adapted to the recommendations made by the Member States of the
European Union because of differences in agricultural practices and landscape and be
continuously updated according to the new available scientific information.
Given that the occurrence of spontaneous inter-specific crossings among oilseed rape,
Brassica rapa and Brassica juncea crops has already been reported, the Group of
Experts recommends that each farmer growing B. napus, B. juncea or B. rapa crops in
the neighbourhood of the transgenic oilseed rape fields is aware of the agricultural
guidelines.
Considering that seeds will be imported and transported and that seed spillage might
occur, feral oilseed rape plants might estab lish and persist outside the fields (e.g. on
roadsides, railways, wastelands, points of import). For this reason, the Group of
Experts recommends that every operator should be informed on the appropriate
management measures to be taken in the case of accidental seed spillage and the
establishment of feral oilseed rape populations. It should also be made clear to whom
these events must be reported. Where seed spillage occurs, the agricultural guidelines
should recommend that the seeds must be swep t or shovelled into sealed containers.
In addition, seed spills should be recorded and monitored in subsequent years in order
to allow appropriate control.
To limit seed spillage during transport and the potential establishment of feral
herbicide-tolerant oilseed rape popula tion, the agricultural guidelines should
recommend transporting the seeds in closed and sealed containers.

To limit the development of multiple herbicid e-tolerant plants through vertical gene
transfer of the herbicide-tolerant trait, the agricultural guidelines should recommend
that:
– the cultivation of transgenic oilseed rape with tolerance to other herbicides in
the same or nearby rotation must be avoided,
– oilseed rape must not be grown on the same land more than one year in four.
– Given that reliance on a single mode of herbicide control exerts a high
selective pressure on the weed population and volunteers, which may lead to
the development of resistance to the concerned herbicide and that rotating
active ingredients reduces resistance de velopment, the agricultural guidelines
should recommend that:
– the glufosinate ammonium must be used in combination with other herbicides
during the same rotation cycle,
– the use of different crops tolerant to glufosinate ammonium must be avoided
in the same rotation cycle,
– integrated weed management systems should be applied.
Given that agricultural weeds become l ess abundant in fields where glufosinate
ammonium is applied and that the environm ental disbenefits of very clean fields
might be judged unacceptable, uncropped fields and/or field margins and untreated
field margin strips might be the only physical habitats to preserve these plants and the
whole chain of living organisms living on and from these plants. Within those fields,
margins or strips, the agricultural weeds and the accompanying fauna may find a
place to survive, provided that the farm ers allow them to. For this reason the
agricultural guidelines should recommend farmers to preserve field margins (hedges,
hedgerows, grassy borders, small woodlots and others), leave field margins unsprayed
and limit herbicide drift. The agricultural guidelines should also recommend farmers
to use band spraying if the negative impact of cleanliness is to be reduced.
To limit any selective pressure of the glufosinate ammonium on plants outside the
field (e.g. herbicide-tolerant feral oilseed ra pe plants, wild relatives in field margins),
the agricultural guidelines should recommend farmers to avoid herbicide drift in field
margins according to the current principles of good agricultural practices.
409. The original proposed Agricultural Guidelines are in EC-63/At.123, 14 January 2003. I
consider the question from the Panel to be ambi guous. The conclusions and recommendations in the
Report of the Group of Experts (RGE) are about improving the original Agricultural Guidelines
(OAG) proposed by the notifier. The conclusion by the Belgian Biosafety Advisory Council (BBAC)
is also about the original Agricultural Guidelines. The stated question may be interpreted to be: do
the conclusions and recommendations in the RGE support the conclusion by the BBAC that the OAG
are "impracticable, hardly workable and hard to control"? Alternatively, the stated question may
interpreted to be: is there anything in the conclusions and recommendations in the RGE that contradict
the conclusion by the BBCA that the OAG are "impracti cable, hardly workable and hard to control"?
I will assume that the stated question is not anythi ng similar to: Are the OAG "impracticable, hardly

workable and hard to control," or are any of the conclusions or recommendations in the RGE
"impracticable, hardly workable and hard to control"? I will address the first possible alternative here.
410. First, it should be clear that the RGE does not contain any conclusion similar to that of the
BBAC, i.e., that the OAG are "impracticable, hardly workable and hard to control."
411. The RGE states
To allow safe and sound integration of MS8, RF3 and MS8xRF3 oilseed rape in
Europe the notifier proposed agricultural guidelines giving guidance to farmers and
seed suppliers. Since the experts considered that the guidelines would be crucial in
limiting vertical gene flow and its potential consequences, it was strongly
recommended that the practical mitigation measures as described in the guidelines
would be followed.
412. This implies that some of the mitigation measur es in the OAG are consider impracticable, as
there would be no need to specify "practical m itigation measures" in the statement if all were
considered practical.
413. The RGE also states
Moreover, it was proposed that the effectiv eness of the proposed risk management
strategies in limiting vertical gene flow (and its potential consequences) and in
preserving farmland biodiversity should be assessed in the post-market monitoring
plan. In addition, an independent farmers' audit to monitor compliance with the
agricultural guidelines should be foreseen on a regular basis.
414. This is an critically important control point th at was lacking in the OAG. This implies that
the RGE considers some of the risk management measures proposed in the OAG to be hard to control.
415. The RGE organizes the recommendations for th e Agricultural Guidelines along routes of
escape and potential adverse effects. The OAG is organized around farm operations. The OAG
emphasizes workability for farmers and seed distri butors, while the RGE emphasizes workability for
attaining the risk management objectives. Obviously , both must be met ultimately, but it is not clear
from the BBAC conclusion what kind of workability is considered.
416. Thus the RGE supports the BBAC conclusion that the OAG are impracticable and hard to
control. It is not clear if the RGE supports the BBAC conclusion that the OAG are hardly workable.
417. In passing, and not really on the point, I ag ree with the BBAC that parts of the OAG are
"impracticable, hardly workable and hard to cont rol," but would prefer to consider these parts as
needing considerable improvement before they ar e acceptable. I also consider some of the
recommendations in the RGE to be "impracticable, ha rdly workable and hard to control," and overall
the Agricultural Guidelines will need additional work so that they accomplish the risk management
goals.
Dr. Squire
418. The question is whether the words "impracticable, hardly workable and hard to control" used
by the Belgian Biosafety Advisory Council are justified on the basis of the report by its Group of
Experts. The Group of Experts does not use this wo rding as such but makes ca utionary remarks that

the guidelines are not sufficient as they stand and shou ld be extended and that there could be practical
problems in successfully implementing them. Examples include –
• "each farmer growing B. napus, B. rapa and B. juncea, in the neighbourhood of the
transgenic oilseed rape field is aware of the agricultural guidelines": this requires that
non-GM farmers need to make themselves aw are of GM issues, and that all farmers
will be cooperative neighbours.
• "every operator should be informed on the appropriate management measures to be
taken in the case of accidental seed spillage …etc.": this in reality is probably
unworkable, since many spillage events w ill go unnoticed, and there are such a large
number of feral populations arising from such events.
• "seed should be swept or shovelled into sealed containers" and "(spillage sites) should
be recorded and monitored in subsequent years": these are likely to be impracticable
in commercial agricultural areas; the measu res would place an intolerable burden on
whoever was considered responsible if that could be decided (the grower, the
transported, the buyer, the roads authority?).
419. Several other of the Group of Experts' recommendations are workable, but the above three
examples would appear to justify the Belgian Safety Advisory Council's choice of words.
Trifolium/Monsanto/Danisco Roundup Ready fodder beet (A5/15)
C/DK/97/01 (EC chronology 64)

Question 21: Given the information before the Pa nel, including the lead CA's positive opinion
(EC 64/At. 29-30) and the conclusions of the EC SCP (EC-64/At.83), did the additional risk
assessment provided by the notifier (EC-64/At.104-105) in response to requests by the Dutch CA
for a theoretical safety assessment address outstanding scientific concerns related to the
potential risks associated with this product?
Dr. Nutti
420. Based on the information provided by the applican t in EC-067/At.1 and the analysis done by
the EC SCP (CDA-35A) and following the Codex Guid elines, my understanding is that the additional
risk assessment provided by the notifier was not necessary in the first place. It does not address
outstanding scientific concern since they are not re lated to the potential risks associated with this
product.
421. It is important to point out that both the EC SCP and the Danish CA overall assessments have
stated that there is no evidence indicating that the use of the fodder beet tolerant to glyphosate with
the purpose to be used as any other fodder beet is likely to cause any adverse effects on human health
and the environment.
Dr. Andow
422. The issue addressed in EC-64/At.104-105, 19 October 1999, was the existence and sequence
of backbone DNA. This issue had not been addressed previous to this date for this transgenic fodder
beet. The notifier indicated that they would provide information documenting that the backbone DNA
was absent, but in the meantime would provide a risk assessment assuming the entire backbone DNA
was present (EC-64/105, 19 October 1999). This risk assessment was not available in any of the
materials before the Panel.

423. The issue addresses the possible activity of back bone DNA in the plant. If it exists and it
becomes active, it could result in unanticipated phenotypes that could lead to adverse effects on
human health or the environment.
424. Thus I conclude that the additional risk assessment would have addressed an outstanding
scientific concern related to the potential risks associated with this product.
425. The request by the Dutch CA was on 4 August 1999, following the launching of the Inter-
Service Consultation on 4 September 1998. The notifier provided additional information on 24
September 1998, 25 September 1998, 5 Octobe r 1998, 13 November 1998 and 5 March 1999.
Counting from the date of the launching of the In ter-Service Consultation, the Dutch CA took 334
days. Counting from the date the notifier last pr ovided additional information, the Dutch CA took
152 days. In either case, the number of days seems like a long time to make the response.
Question 22: Given the information before the Pa nel, including the conclusions of the SCP's
risk assessment, was the information requested by the Italian CA in March 2000 (EC-64/At.116)
necessary to ensure that conclusions of the safety assessment were valid?
Dr. Nutti
426. Based on the information provided by the applican t in EC-067/At.1 and the analysis done by
the EC SCP (CDA-35A), I understand that there was no need, as far as food safety is concerned, for
the request made by the Italian CA (EC-64/At.116) in order to ensure that conclusions of the safety
assessment were valid.
Dr. Andow
427. The information requested by the Italian CA (EC-64/At.116, 14 March 2000) is as follows.
Referring to the conclusion of the last meeting of the Regulatory Committee under
Directive 90/220/EEC, Italy would be very pleased to receive from the notifier more
information concerning the transfer and the recombination of genes in natural
conditions, on the efficacy of the use of the Roundup herbicide and on the evaluation
of the possible crossing with spontaneous species.
428. Issues related to the transfer and recombinati on of genes in natural conditions and to the
evaluation of possible crossing with spontaneous (w ild?) species were addressed in the SCP Opinion
(EC-64/At.83, 23 June 1998) and were addressed in the original notification.
429. It is possible that the Italian CA believed these assessments to be inadequate. If so, the CA
should have indicated this in the question. However, as discussed below, this is unlikely.
430. Issues related to the efficacy of the use of glyphosate were not addressed in the SCP Opinion,
but I fail to understand how such issues relate to a risk assessment, and the Italian CA provides no
guidance. This question was NOT necessary to ensu re that conclusions of the safety assessment were
valid.
431. In the response to the Italian CA (EC-64/At.119, 12 July 2000), the notifier states that there
are no new data in the document, only clarifications. Given that the Italian CA did not follow up on
this response, I must assume that the response satisfied the Italian CA's concerns.

432. Thus, the issues raised by the Italian CA had been addressed in previous documents. Based
on the detailed responses from the notifier in EC- 64/At.119a, the question on possible crossing with
spontaneous species was NOT necessary, but the ques tion on transfer and recombination in natural
conditions was necessary to ensure that conclusions of the safety assessment were valid.
Question 23: Given the information before the Panel, including the conclusions of the lead CA's
positive opinion (EC-64/At.29-30) and EC SCP opinion, did the additional information provided
by the notifier in response to requests by the Du tch CA and by the UK for additional data
regarding molecular characterization address outstanding scientific concerns related to
potential risks associated with this produc t (EC-64/At.113 and 119)? Was the molecular
characterization provided by the applicant suffi cient to enable the Dutch CA and the SCP to
conduct a risk assessment and to support the conc lusions reached regarding the safety of the
product?
Dr. Andow
433. The issue addressed in EC-64/At.104-105, 19 October 1999, was the existence and sequence
of backbone DNA. This issue had not been addressed previously for this transgenic fodder beet. The
notifier indicated that they would provide in formation documenting that the backbone DNA was
absent, but in the meantime would provide a risk assessment assuming the entire backbone DNA was
present (EC-64/105, 19 October 1999). This risk assess ment was not available in any of the materials
before the Panel. The notifier provided the Dutch CA with the molecular evidence in EC-64/At.113,
23 February 2000.
434. This issue addresses the possible activity of backbo ne DNA in the plant. If it exists and it
becomes active, it could result in unanticipated phenotypes that could lead to adverse effects on
human health or the environment.
435. Thus I conclude that the additional informa tion addressed an outstanding scientific concern
related to the potential risks associated with this product.
436. The UK CA also addressed the same issue to th e notifier, apparently unaware that the issue
had been raised by the Dutch CA and addressed alrea dy by the notifier. The notifier clarified this in
EC-64/At.119, 12 July 2000.
437. The molecular data was included with a summary and transmittal letter from the notifier on
23 February 2000. I will not comment on the sufficiency of these data.
Monsanto Roundup Ready cotton (RRC1445)
C/ES/97/01 (EC chronology 66)

Question 24: Does the information before th e Panel, including the full application
(EC-66/At.3-12) and the EC 's SCP opinion (EC-66/At.43), provide scientific support for the
objections raised by EC member States (EC-66/At.57) concerning the adequacy of the
monitoring plan, the antibiotic resistance marker genes, and herbicide residues?
(a) What criteria can be used to judge if the final monitoring plan submitted by
Monsanto in January 2003 was complete?

Dr. Andow
Scientific support for objections:
438. This response addresses only issues related to the adequacy of the monitoring plan, and does
not address issues related to antibiotic resistance marker genes and herbicide residues.
439. The lead CA reached the following conclusions (EC-66/At.05, Submission of lead CA to
COM, 19 Nov 1997, Overall Assessment).
2. From the risk assessment it is conc luded that there is no reason to suppose
that the harvest and handling of RRC Line 1445 tolerant to glyphosate herbicide,
have adverse effects on the environment and human health.
9. The dossier was considered by Nationa l Commission on Biosafety. The main
aspects considered in the risk assessment were:
– Capacity to survive, establish and disseminate.
– Potential for gene transfer.
– Products of expression of inserted sequences.
– Phenotypic and genetic stability.
– Pathogenicity to other organisms.
– Potential for adverse effects to humans.
10. The National Commission on Biosafety considers that, for the considered
uses, there is no significant difference as fa r as environmental and human health risks
related to other cotton.
16. Finally, as far as potential effects for non-target organisms is concerned, CP4
EPSPS protein is broadly present in nature. Therefore organisms which feed with
plants and microorganisms are exposed to this protein. Studies with birds feeded
with RRC line 1445 cottonseed meal have been conducted and no significative
difference of this feeding was detected. Moreover, EPSPS protein exist in nature and
is considered non-toxic to animal species. On the other hand, cotton is a unique crop
that mammals and other species which consume vegetation avoid feeding on the plant
due to both gossypol in the plant and the morphology of the plant.
440. These excerpts show that the risk assessment da ted 19 November 1997 by the NCB did not
consider indirect effects on non-target organisms, long-term or spatial scale effects on non-target
species, effects associated with changes in the cropping system or the evolution of resistance in weeds
to glyphosate. The NCB does consider whethe r there are any significant differences between
anticipated risks from RRC 1445 compared to conventional cotton. Although the NCB concludes that
there are no significant differences (point 10), they do not provide a scientific argument that this
conclusion extends to the potential effects not cons idered in the risk assessment. Because the NCB
considered that the risks they assessed were inconsequential, the NCB did not propose any monitoring
plan.

441. The SCP provided the following Opinion (EC-66/At.43, 14 July 1998. Opinion of the
Scientific Committee on Plants).
6.3.3. Safety to non-target organisms: Exposure of non-target species to seeds can
be considered very low, due to the morphology of the boll. Feeding studies with
birds (seeds) and mammals (both proteins) indicate very low toxicity of the proteins
which also occur ubiquitously in the environment in plants and microorganisms.
Field studies on agronomic performance showed equivalent susceptibility of line
RRC 1445 and non-modified varieties to diseases and insect pests.
6.3.4. Resistance and tolerance issues: Any selective advantage of cotton line RRC
1445 is restricted to cases where no herbicide other than glyphosate is used on early
stages of cotton. Under normal applicati on rates, the introduced glyphosate-tolerance
is effective up to the 4-leaf stage only. Other herbicide, cultivation of rotational crops
or winter conditions will kill both modified and non-modified plants. Volunteers
should be dealt with by standard agricultural practice except that glyphosate should
not be used.
442. These excerpts show that they did not consider indirect effects on non-target organisms, long-
term or spatial scale effects on non-target species, effects associated with changes in the cropping
system or the evolution of resistance in weeds to glyphosate. Because the SCP considered that the
risks they assessed were inconsequential, the SCP did not propose any monitoring plan.
443. Sweden commented as follows (EC-66/At.57, 26 April 1999. Consultation of the Committee
by Written Procedure).
Sweden has, in different EU contexts and in earlier statements, put forward the view
that herbicide tolerant crops should not be placed on the market until the long-term
effects of herbicide tolerant crops on the environment have been better analysed.
Common principles for evaluation and monitoring of the risks connected to the
cultivation of herbicide tolerant crops should be established.
444. Sweden suggests that long-term effects shoul d be studied prior to marketing and that
principles for monitoring risks related to the cu ltivation of herbicide tolerant crops should be
established. Sweden is saying that effects associ ated with changes in the cropping system should be
considered both in the assessment and monitoring of risk. The scientific literature at that time
indicated that long-term effects frequently elude de tection when assessed on short time scales, so one
of the only reasonable ways to address these kinds of effects is via monitoring. It does not seem
appropriate to require long-term experiments for risk assessment because that could delay the process
by a decade, so monitoring is a possible alternative.
445. The United Kingdon commented as follows (EC-66/At.57, 26 April 1999. Consultation of
the Committee by Written Procedure).
We would also like to draw to the attenti on of other Member states where GM cotton
might be widely grown, that this cott on line may have a negative impact on
biodiversity arising from changes in the way the herbicide tolerant crop is managed.
446. The United Kingdom is drawing attention to spa tial scale effects and effects associated with
changes in the cropping system. The scientific literatu re at that time indicated that large-scale effects

frequently elude detection when assessed at smaller spatial scales. One of the reasonable ways to
address these kinds of effects is via monitoring.
447. The information in the full application (EC-66/At.3-12) and the EC's SCP opinion
(EC-66/At.43) do not provide scientific support for the objections raised by EC member States
(EC-66/At.57) concerning the adequacy of the mon itoring plan. This is because the scientific
information in the full application and the EC-SCP opinion do not address the scientific basis for the
objections raised by the EC member states concerning the adequacy of the monitoring plan.
448. The objections of the member States (EC-66/ At.57) concerning the adequacy of the
monitoring plan raise new scientific issues that we re not assessed in the full application (EC-66/At.3-
12) or the EC's SCP opinion (EC-66/At.43). There is no monitoring plan proposed in either the full
application (EC-66/At.3-12) or the EC's SCP opi nion (EC-66/At.43), so the EC member States'
objections can only be interpreted that a monitoring plan may be or is necessary.
449. The objections of the member States (EC-66/At.57) concerning the adequacy of the
monitoring plan are scientifically justifiable. The EC member States raise specific scientific issues
that can be addressed in a monitoring plan. Howeve r, the necessity of a monitoring plan cannot be
determined from these objections.
450. It should also be noted that the objections of the member States (EC-66/At.57) concerning the
adequacy of the monitoring plan are not all clearly stated as monitoring issues, and the objections do
not provide clear guidance to an applicant for how to fully respond.
451. Excerpt from EC-66/At.64. Letter from Span ish CA to Monsanto requesting additional
information regarding the monitoring plan, 1 January 2003.
Plan de seguimiento
Se deberán concreter y desarrollar aquello s aspectos susceptibles de ser sometidos a
herbicida Glifosato y sus potenciales efect os a largo plazo (incidencia de malas
hierbas, resistencias, etc), así como cualquier otro efecto relacionado con cambios en
las prácticas agrícolas convencionales.
452. I believe the Spanish CA is requesting that the monitoring plan address potential effects of
large spatial scale (for example, weed incidence, resi stance) and in relation to changes in agricultural
practice. These echo the comment of the UK (EC-66/At.57) and some of the comment of Sweden
(EC-66/At.57).
453. The UK Farm Scale Evaluations (FSE) 65 of GMHT crops published in 2003 indicate that one
anticipated effect of GMHT crops is an alterati on in weed populations and communities compared to
conventional production. The precise alteration probably depends on the herbicides used on the

65 Andow 2003, Nature Biotechnology, 21, 1453-1454. Brooks et al. 2003, Phil. Trans. R. Soc. Lond.
B, 358, 1847-1862. Champion et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1801-1818. Firbank 2003, Phil.
Trans. R. Soc. Lond. B, 358, 1777-1778. Haughton et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1863-1877.
Hawes et al. 2003, Phil. Trans. R. Soc. Lond. B, 1899-19 13. Heard et al. 2003, Phil. Trans. R. Soc. Lond. B,
358, 1819-1832. Heard et al. 2003, Trans. R. Soc. Lond . B, 358, 1833-1846. Roy et al. 2003, Phil. Trans. R.
Soc. Lond. B, 358, 1879-1898. Squire et al. 2003, Phil. Trans. R. Soc. Lond. B, 358, 1779-1799.

GMHT crop and the conventional crop. Nearly a ll other adverse effects on non-target species would
likely follow from these changes in weed populations and communities. Changes to weed populations
and communities have not been reported in GMHT cotton on a scale similar to the UK-FSE trials.
These studies follow on a concern published in 2000 that GMHT crops could adversely affect
skylarks in the UK. 66 Although none of these studies existed at the time of the decisions by the NCB
or the SCP, they partially validate some of the hypothetical concerns expressed by the various
countries, especially related to spatial and temporal scale.
454. Considerable theory and evidence indicates that another anticipated effect of GMHT crops is
the development of resistance to glyphosate in some weed populations. 67 Resistance to glyphosate in
a species of morning glory, which is a weed found in GMHT cotton in southeastern US, has been
recently reported.
68 However concerns about resistance have been widely recognized for some time
before 1998.
455. Potential long-term effects of GMHT cotton have not yet been identified and verified
scientifically.
456. The Member States submitted questions to the notifier on 22 February 1999. The notifier
submitted materials to the SCP on 6 July 1998. The SCP returned an Opinion on 14 July 1998. The
notifier submitted additional information that was circ ulated on 20 November 1998. After this follow
3 decisions to postpone the decision-making pro cess. Depending on which dates are used, the
Member States used either 94 or 231 days to respond to the most recent submission by the notifier.
The shorter time frame is reasonable, especially as it spans the winter holiday season when most
offices are closed for long periods of time. The longer period of time seems excessive to formulate
and provide the responses to the notifier.
Criteria to judge monitoring plan:
457. The specific purpose of monitoring should be clearly stated. This purpose should be linked to
the management of some risk. This linkage ties monitoring to risk management, and delimits the
monitoring endpoint. Although Annex VII.A of Directive 2001/18/EC provides a statement about the
objective of a monitoring plan, this statement is not specific enough as the purpose of any particular
monitoring plan. The plan should be more closely linked to actual potential risks, as indicated in
Annex VII.C.1-2 of Directive 2001/18/EC.
458. For example, there are several possible pur poses for monitoring herbicide resistance 69 in
weeds, including:

66 Watkinson et al. 2000, Science, 289, 1554-1557.
67 See www.weedscience.org, which lists all occurrences of weeds resistant to a herbicide worldwide,
including cases of resistance to glyphosate.
68 Baucom and Mauricio 2004, PNAS 101: 13386-13390.
69 Here I will not distinguish between resistance and to lerance, and will refer to both as "resistance."
Baucom and Mauricio (2004, PNAS 101: 13386-13390) distinguish tolerance from resistance, basing the
distinction on Painter's (1958, Annu. Rev. Entomol. 3: 2 67-290) framework. They defi ne tolerance as a trait to
compensate from herbicide damage and resistance as a tra it that prevents herbicide damage. Painter, however,
considered both of these to be a part of the concept of resistance, and sugge sted that resistance is divided into
"tolerance", "antibiosis" and "nonpreference." Antibiosis and nonpreference correspond to the narrow definition
of "resistance" advanced by Baucom and Mauricio. I adopt the broader definition of resistance stemming from
Painter's work.

• Purpose R1. Document/Measure the occurren ce of herbicide-resistant weeds. This
would provide information prior to any control failures that could be used to alter the
use of the herbicide or herbicide-tolerant 70 crop to delay or a void higher levels of
resistance in the weed. Because this o ccurs before any control failures, it would
provide time to develop a reasoned response to the threat. This would be particularly
advantageous to prolong the useful life of the herbicide in question, especially if it
had replaced herbicides that caused greater damage to the environment.
• Purpose R2. Document/Measure the occurre nce of a weed problem that cannot be
controlled by the herbicide. This could provide information of localized control
failures that could be used to alter the use of the herbicide or herbicide-tolerant crop
to delay or avoid higher levels of resist ance in the weed. Because some control
failures will have occurred, it may be n ecessary to mobilize a rapid response to
prolong the useful life of the herbicide.
• Purpose R3. Document/Measure the widespread occurrence of a weed problem that
cannot be controlled by the herbicide. This would provide information of widespread
occurrence of the risk, and would be an indi cation that the risk management practices
had failed. One crucial use of monitoring is to determine when a predefined point of
failure is reached.
459. Similarly there are several possible purposes for monitoring changes in weed populations and
communities, including:
• Purpose W1. In the UK, there has been a concern that GMHT crops would adversely
effect the skylark, a desirable species.
71 Skylarks feed on weed seeds, so monitoring
of weed populations and communities could be aimed to monitor the abundance of
food for skylarks. This approach could be extended for any other non-target species
of concern.
• Purpose W2. There is a possibility that "unanticipated" or "unexpected" adverse
effects could follow from changes in weed populations and communities. Monitoring
these weeds would be a prelude to discovery of unanticipated or unexpected adverse
effects.
460. In addition, there are several possible purpo ses served for monitoring for "unexpected" or
"unanticipated" effects, including:
• Purpose U1. Document the geographic and temporal pattern of use of the GMHT
crop so that there is a database availa ble that would enable retrospective
epidemiological-like investigations should an "unanticipated" or "unexpected" effect
be observed.
72
• Purpose U2. Train personnel who normally visit agricultural fields and natural areas
near agricultural fields to be aware of the possibility that agriculture generally and
GMHT crops specifically may have "unexpected " or "unanticipated" effects. Inform
these people of the occurrence of GMHT crops . Training should include information

70 The use of "tolerance" in this context is different from the use of tolerance as defined either by
Painter (1958) or Baucom and Mauricio (2004). In actuality, these GMHT (genetically-modified herbicide-
tolerant) crops are not damaged by the herbicide, unless the herbicide is applied inappropriately.
71 Watkinson et al. 2000, Science, 289, 1554-1557.
72 This was recommended by an NRC committee (N RC 2002, Environmental e ffects of transgenic
plants: The scope and adequacy of regulation. NAS Press, Wash, DC). It finds resonance in Annex VII.C.4 of
Directive 2001/18/EC, but the Annex is not as clear as the recommendations in NRC 2002.

about the potential mechanisms by which such effects could possibly arise. For
example, "unanticipated" effects of weed co mmunity shifts should receive attention.
This training will provide the many eyes needed to notice possi ble changes in the
environment.73
• Purpose U3. Document changes in agricultu ral practices associated with the use of
GMHT crops. There is a possibility that "unanticipated" or "unexpected" adverse
effects could follow from changes in agri cultural practices. Monitoring these
practices could be a prelude to discovery of unanticipated or unexpected adverse
effects.
461. Clear specification of the purpose of monitoring is probably the most critical step in
developing a useful monitoring system.
Monitoring must have a sequel
462. The act of monitoring should not be an end in itself. The information gathered during the
monitoring activities should be used to make a respon se. This response should be related to the risk,
including activities ranging from those to better estimate the risk to those activities to avoid, mitigate
or tolerate the risk.
74 Specification of a response is probably the second most critical step in
developing a useful monitoring system. Although Annex VII.C.6 of Directive 2001/18/EC states that
a response should be considered, it does not require a response.
Action Trigger
463. In addition to a sequel, a monitoring system mu st have a well-defined action trigger. An
action trigger is a criterion or set of criteria, which if met (or exceeded) according to the information
from monitoring, would require taking the response actions. Without a clear action trigger, it could
become quite difficult to determine when monitoring would precipitate a response.
464. Important logistical issues include: (1) who will monitor, (2) who is responsible for
monitoring, (3) who handles and synthesizes the mon itoring information, (4) who verifies the quality
of the monitoring, information obtained from monito ring and synthesis of this information, (5) to
whom are monitoring results reported and at what frequency, (6) how will the monitoring effort be
sustained, (7) how can monitoring be conducted in a cost-effective manner. Most, but not all, of these
logistical issues are recognized in Annex VII.C.3 and 5 of Directive 2001/18/EC.
465. Important methodological issues include: (1) what 'endpoint' will be monitored, (2) what
frequency will monitoring be conducted, (3) what will be the spatial density (grain) of monitoring, (4)
can monitoring be stratified by potential risk. A fe w of these issues are recognized in Annex VII.C.3
of Directive 2001/18/EC. The endpoint of monitoring is what is actually measured and/or estimated
by the people conducting the monitoring. For exampl e, for weed resistance it could be the frequency
of resistance genes or resistant phenotypes in the weed population; for non-target effects, it could be
total weed biomass, production of weed seeds that ar e normally consumed by skylarks, and so on.
Choice of endpoint can enable or restrict possible responses. The frequency of monitoring will
depend on several factors, including the anticipat ed response. If the response involves a series of

73 This was recommended by an NRC committee (N RC 2002, Environmental e ffects of transgenic
plants: The scope and adequacy of regulation. NAS Press, Wash, DC). It finds resonance in Annex VII.C.3.2 of
Directive 2001/18/EC, but the Annex is not as clear as the recommendations in NRC 2002.
74 This point has been made emphatically in NRC 2001. Ecological monitoring of genetically modified
crops. National Academy Press, Washington, DC. 45pp.

processes that would take several years to comple te, it might be appropriate to increase monitoring
frequency so that the monitoring process does not in troduce additional delays in responding to a
potential threat. The grain of monitoring will be depend on several factors, including the expected
grain of the effect being monitored. For example, if regional weed resistan ce is the concern, then
monitoring can be done at a regional scale. If weed resistance on farms is the concern, then
monitoring will probably need to occur at a finer grain. Weed community shifts might be done on
several spatial scales. If the adverse consequences are focused on bird species, regional monitoring
may be sufficient. If they are focused on less mob ile species, then a finer grain will be necessary.
Finally, it will be essential that monitoring is stratified so that it occurs at places and times most likely
related to possible risks. Monitoring is costly a nd monitoring resources must be allocated efficiently.
For example, monitoring for unanticipated effect s could be stratified according to the geographic
distribution of the GMHT crop. Monitoring coul d occur mostly or only at places where the GMHT
crop is used considerably, with a threshold inte nsity of usage determining the distribution of
monitoring effort.
466. Monitoring for unanticipated or unexpected effects. The first step in developing a monitoring
system for these possible effects is to specify clearly wh at effects, if any, are anticipated or expected.
Without this clear statement, it will not be possible to determine if any observed effect is
unanticipated or unexpected. For example, if no adverse effects ar e anticipated or expected, then it
must be granted that any subsequently observed effect must be considered unanticipated or
unexpected.
467. General critique of proposed monitoring plan (EC-66/At.62). (1) Specific purpose of the
proposed monitoring plan is to monitor for unanticip ated or unexpected effects. It does not address
concerns associated with weed resistance or chang es in weed populations and communities, as
indicated by the EC officials. The purpose of th e monitoring for unanticipated or unexpected effects
is not entirely clear. Consequently it is difficult to know how the proposed monitoring will enable the
identification of any unanticipated or unexpected e ffects. (2) The proposed response is to conduct a
scientific evaluation of the observed potential unanticip ated effect to confirm that it is in fact an
unanticipated effect.75 While it is stated that the response should be proportionate to the risk, it is not
clear what procedure and standard will be used to determine a proportionate response. Second is
unclear who is responsible for conducting the scien tific evaluation and how to ensure that this
information will be collected in a timely fashi on, who will have access to the information, who
determines the adequacy of the design of the evalua tion and so on. (3) The triggers for initiating a
scientific evaluation are not clear. It would be folly to initiate a scientific evaluation of all possible
observations of possible adverse effects. Ho w will these be screened to trigger a scientific
evaluation? When does the scientific evaluation provide sufficient evidence to trigger remedial
action? These issues are not resolved in the prese nt proposed monitoring plan. (4) The logistical
issues are better specified than the other component s of the proposed plan. Important weaknesses in
the proposal include lack of specification for how the seed supply and distribution network will be
linked to the monitoring procedures, that key exte rnal networks are not committed to monitoring,
procedures to verify the quality of monitoring, in formation from monitoring and synthesis of this
information, short time span and long intervals for reporting, and so on. (5) Methodologically, the
proposal has several important weaknesses. No e ndpoint is specified and it is not clear how people
who might be monitoring would recognize an unanticipated effect. In this case these people should be
informed that any adverse effect mu st be considered an unanticipated effect, and should be reported.
The frequency and grain of these obser vations is unclear. There seems to be little stratification of
effort by potential risk.

75 Page 4, Evaluation of potential adverse effects.

Dr. Squire
468. The EC's SCP opinion is generally positive about the application, whereas the EC member
states raise several objections. The issue of antibiotic resistance was considered in the SCP's opinion
(EC-66/At.53) and found not to pose risk, but ther e is now widespread perception that antibiotic
resistance should not be introduced through GMHT products. The basis of objection by some member
states is that general effects of HT cropping on farmland habitats are uncertain, and this applies
whatever the species or the land area it occupies. However, the context for this biotech product in
cotton is very different from the context of those products which are varieties of oilseed rape. Cotton
occupies a very small area in Europe and does not present potential problems of the type associated
with oilseed rape or even maize or beet (Table 1). This notwithstanding, and as in other instances,
unless criteria can be given, from both the proposer and objector as to what is a desirable or
acceptable comparator, then progress with the discussion is impossible, as it became in this instance.
469. There appeared to be no monitoring plan in the original proposal, and in the 2003 document,
the relevant section indicated that little specific m onitoring was necessary b ecause of the low risk.
Monitoring the effects on habitat would be feasibl e but suitable criteria on which to base monitoring
have not been proposed or agreed by either side.
Amylogene starch potato
C/SE/96/3501 (EC chronology 67)

Question 25: Given the information before the Panel, including the application (EC-67/At.13-
44), and additional information provided by the notifier (EC-67/At.51, 57, 61-63, 75-83, 87,
92-93, 94-95, 101, and 103), was the information regarding molecular characterisation, toxicity,
protein analysis, animal feed trials, effects on non-target organisms, bleomycin resistance, and
substantial equivalence requested by the Scient ific Committee for Food (SCF) (EC-67/At.84-86,
96, 98, 100, 102, 104, 105 and 106), necessary to support a valid safety assessment?
Dr. Nutti
470. Based on the information provided by the applicant in EC-067/At.13-44 and the analysis done
by the EC SCP and following the Codex Guidelines, it is clear that the information required was
necessary to support a valid safety assessment. The no tifier failed to provide information on several
fundamental issues concerning safety such as substantial equivalence, toxicity, protein analysis and
animal feeding trials.
Dr. Andow
Necessity for valid safety assessment:
471. In this answer I will address only the non-target organism part of the question. The transgenic
potato produces elevated concentrations of amyl opectin by blocking starch synthesis and also
expresses nptII. Based on molecular analysis there are multiple copies of the transgene in the plant.
472. EC-67/At.84, 4 February 1999, First request from SCP to Amylogene for clarification:
In conclusion, I would like to inform you that the Committee will only consider at
this stage additional data that is of a non-confidential nature. In this respect, I would
be obliged if you would include a statement to this effect in the covering letter with
any data you may wish to submit in r esponse to the Committee's questions. The

reason for this is that it is not possible at this stage to repeat the notification
procedure, which provides for agreement between the notifier and the competent
autority on confidentiality (Article 19 of Council Directive 90/220/EEC). However
should you consider that certain of the data requested by the SCP is of a confidential
nature and that you would prefer not to submit the data without obtaining a
confidential status, the SCP will then complete its evaluation on the basis of the
available data.
76
8) No data are provided on the safety of modified crops to non-target organisms.
No risk assessment has been made.77
3.5. Environmental Aspects
3.5.1. Geographical relevance of data
Wherever possible data should be provi ded from field experience in those
geographical regions where the GM plant will be grown commercially to reflect
relevant meteorological, soil and agronom ic conditions. Where data from field
studies on other continents are supplied, the notifier should submit a reasoned
argument that the data is applicable to Europe. Bridging studies may be particularly
useful.
3.5.2. Impact on non-target organisms
Clear and well-defined risk assessments should be carried out for each of the different
functional environmental compartments that are exposed to the GM plant; this will
depend on the specific crop and if any parts of it will remain in the environment after
harvest. For example, exposure should be estimated to soil organisms and function
(e.g. earthworms, micro-organisms, leaf litter breakdown), non-target arthropods
(including pollinators, beneficial arthr opods), grazing birds and mammals or, if
appropriate, the aquatic environment. Th is risk assessment should take account of
where in the plant the inserted genes are expressed and the consequent exposure of
non-target organisms. The assessment s hould also address the fate of the expressed
substance in those environmental compartm ents where they are introduced and may
cause exposure of non-target organisms (e.g. in soil after the incorporation of plant
material). Data on the comparative su sceptibility of the GM plant to pests and
diseases compared to the non-modified plants are useful together with observations
on agronomic performace during greenhouse and experimental release trials.78
473. The SCP indicated that no non-target assessment had been conducted, and provided guidance
(3.5) for completing such an assessment. At this time, possible risks could arise from the transgenic
plant or processing wastes, including fruit juices and fruit water.
474. EC-67/At.86, 12 July 1999, Draft opinion of SCP
6.3.3. Safety to Non-Target Organisms : There are no data on safety of the
modified crops to non-target organisms. However, the results of growing trials

76 Page 1. All page references in this answer refer to the page number in the relevant pdf file.
77 Page 3.
78 Page 9.

suggest neither greater susceptibility nor great er resistance to pests and diseases than
non-GM potato lines. In view of this and the equivalent composition of the modified
potato line, it is considered that no adve rse effects on non-target organisms would be
expected from cultivation of the GM high amylopectin potato line EH92-527-1.79
475. The SCP draft opinion reverses their previous opinion that there has been no risk assessment
for non-target effects. In what follows, I lay out th e findings that appear to lead to both the original
opinion and the reversal, and evaluate the scientific basis for each.
476. EC-67/At.77 1 April 1999, Amylogene provides SC P working group with the full application
as well as additional data. This is copy of EC- 67/At.14 7 May 1998, Summary statement of CA of
Sweden
Environmental effects of the genetically modified potato clone (EH92-527-1)
In laboratory and field trial studies no differences in behaviour of clone EH92-527-1
have been observed compared to its parental variety. The genetically modified clone
is not considered to have increased surviv ability or competitiveness in agricultural or
natural environments. The analysis carried out to investigate whether the modified
clone displayed any increase or decrease in frost tolerance did not indicate any such
changes. The parental variety has a low flowering frequency and thus a low seed
production, and with respect to this the modified clone does not display any change.
Compared to the parental variety, no data indicate that the genetically modified clone
possess any alteration in its potential to sp read its genes, neither through sexual
crosses not horizontally to sexually in-c ompatible species. No unexpected
(pleiotropic) effects resulting from the modi fication of the clone in question have
been identified. In conclusion, the modified clone does not exhibit any differences in
environmental behaviour as compared to the parental variety (cv. Prevalent), and is
therefore not considered to pose any risk to the environment.
80
477. The summary risk assessment statement of the CA of Sweden on 7 May 1998 does not
mention specifically any non-target risks.
478. EC-67/At.79 (also EC-67/At.17) Part I B.7 81, Potentially significant interactions of the plant
with other organisms than plants in the ecosystem where it is normally grown, including information
on toxic effects on humans, animals and other organisms . Here the notification suggests that changes
in glycoalkaloids or nitrates may be causes of non-target effects. The extant literature is reviewed for
effects of glycoalkaloids and nitrates on organisms. Mammals, birds, insect pests of potato, nematode
pests of potato, pathogenic fungi of potato, pathogenic bacteria of potato and infectious viruses of
potato are listed. There is only citations of the toxi c effects of glycoalkaloids to some of the listed
species. No citations to effects of nitrate are mentioned.

79 Page 6.
80 Page 5.
81 Page 15 ff.

479. EC-67/At.79 (also EC-67/At.17) Part I D.9
9. Potentially significant interaction with other, not modified organisms
In potatoes normally existing toxic and anti- nutritional substances are glycoalkaloids
and nitrates. The interaction with other orga nisms has been described in Part 1, B.7..
In D.7, in Annex 8 and in Part 3 it is show n that the contents of these substances have
not been changed as a consequence of the modification. On that account, the
modified potato clone, EH92-527-1, does not affect other organisms in another way
than what the unmodified recipient cultivar, Prevalent, does.
The modified clone, EH92-527-1, is characterized by two changes compared to the
recipient clone. Those are resistance to ka namycin and the lack of amylose. Because
of the latter, the content of amylopectin has been increased to >98%.
82
There are no reasons what so ever to s uppose that the changed starch composition
(>98% amylopectin) should cause any danger for other non-modified organisms,
including humans. There are already pl ants, in which this trait has occurred
spontaneously or by induced mutations, e.g. maize and barley. Further more
"amylopectin starch" can be produced from normal potato starch by chemical
treatment. No un-desired side-effects have been observed (Annexes 5 and 7).
83
480. Specifically amylopectin starch o ccurs in large quantity in the so-called waxy maize varieties
and is also found in quantity in some rice, barley and sorghum varieties.
481. EC-67/At.79 (also EC-67/At.17) Part III D
D. THE RISK THAT THE PLANT CAN BE MORE SUSCEPTIBLE TO
PREDATORS AND DISEASES.
The establishment and development of plants from seed of the clone EH92-527-1 has
been studied in comparative trials in 1995, and in observation trials in 1993, 1994,
1995 and 1996. In all trials it has been shown that the establishment and
development of plants are equal to the recipient clone both above and below ground.
During the growing season, both the modified potato clone and the recipient clone
were exposed to attack by insects and diseases, mainly potato late blight. In order to
avoid such damages, treatments were applied according to the requirements of the
recipient clone. There were no observations indicating that the transgenic clone is
more (or less) susceptible to any pest or di sease than the recipient variety. (Annexes
2, 10, 13, 14, 15 and 20.)
84

82 Page 28.
83 Page 31.
84 Page 45.

482. EC-67/At.80 (also EC-67/At.17) Part III E
E. THE POSSIBILITIES FOR NEGATIVE ECOLOGICAL AND OTHER
EFFECTS WHICH ARE NOT RELATED TO WEED CHARACTERS
Such effects have not been observed during any of the trials. It is not expected that
such effects should occur as a consequence of the modifications present in EH92-
527-1 (Annexes 2, 10, 13, 14, 15, 20).
85
483. It should be clear that the relevant information is in Annexes 2, 10, 13, 14, 15, and 20.
• Annex 2. The Swedish Seed Testing and Certification Institute: Statement on variety
distinctness
• Annex 10. Observations in practical cu ltivation of conventional Prevalent and
transgenic Prevalent EH92-527-1, 1994- 1995, and in laboratory freezing tests
1996/1997
• Annex 13.Report about field trial with transgenic potatoes 1993
• Annex 14. Report about field trials and seed production with transgenic potatoes 1994
• Annex 15. Report about field trials and practical cultivation with transgenic potatoes
1995
• Annex 20. Report about field trials and practical cultivation with transgenic potatoes
1996
484. Annex 2 is missing from the materials. This annex is the Swedish Seed Testing and
Certification Institute: Statement on variety distinctness, so is unlikely to have information relevant to
biosafety risk.
485. Annex 10 contains detailed information about 1994 and 1995. Annexes 14 and 15 provide
only summary information related to potato seed pr oduction, and have no info rmation relevant to
biosafety. Annex 20 provides only summary inform ation related to potato seed production, and has
no information relevant to biosafety. Annex 13 provides detailed information about 1993 results.
However, only four clones were untransformed standards and 79 clones were transformed. This study
was too small a scale to provide reliable information related to biosafety.
486. EC-67/At.81 (also EC-67/At.27), Annex 10. The notification concludes as follows:
Conclusions of practical cultivation in 1994 and 1995
The potato clone Prevalent 92-527-1 has not differed from conventional Prevalent in
the cultivations observed.
– Normal morphology
– Similar reactions to chemicals

85 Page 1.

– Similar resistance to diseases
– Similar sensitivity to frost in early summer as well as in fall
Three comparative trials with comparisons of yield and starch contents were made in
1996 with conventional Prevalent and the clone 92-527-1 (Annexes 8 and 20).
Investigations regarding resistance to nematodes, performed at the Agriculture
Laboratory in Lyckeby, revealed identical resistance to Globodera rostochiensis,
pathotype Ro1Ro4, in both clones.86
487. I will show that the conclusion that the clones are similar in resistance to diseases has not
been scientifically demonstrated.
488. EC-67/At.81 (also EC-67/At.27), Annex 10. Summa ry of data. Five plantings are reported,
two in 1994 and 3 in 1995. At the 1994 Skara planti ng, there were bad growth conditions and yield
was low and seed quality was bad. Fields were treated with fungicide and no late blight occurred. At
the 1994 Norra Sunderyn planting, growing conditions were good, but there was no late blight in the
field. At the 1995 Sofielund planting, growing conditions, plant development and yield were
comparatively bad, late blight was similar betw een transgenic and non-transformed varieties, and
there were no attacks of pests or parasites. At the 1995 Habo planting, soil structure was damaged,
yield was low and late blight was controlled by no rmal fungicide applications. At the 1995 Norra
Sunderbyn planting, growing conditions were good, but no attack by late blight or other diseases were
observed.
489. To show that the clones are similar in resist ance to diseases, the diseases must be present and
the plants must be healthy. Of the five plantings with sufficient informati on to evaluate (paragraph
488), none meet these two conditions. If we co nsider only those plantings with disease present,
disregarding the quality of the plants, one planting (1995 Sofielund) would suggest that late blight
resistance was similar.
490. It can be concluded that there is no evidence in the written record to support the statement in
the draft SCP opinion (EC-67/At.86) that "the resu lts of the growing trials suggest neither greater
susceptibility nor greater resistance to pests and di seases than non-GM potato lines." Indeed, even if
the statement is restricted to refer only to late blight, it is not supported by the evidence. It is possible
that the notifier presented information verbally to th e SCP on this issue that is not recorded in the
written record, but in this case, the SCP should have mentioned this.
491. Thus it can be argued that a request for additional non-target information was
scientifically justified in 1999 and remains justifiable even today.
492. However, it is useful to consider if there is any evidence that points to the possible existence
of non-target effects. The notification identifies two possible routes by which non-target effects could
result: changes in glycoalkaloids or nitrates. Ne ither of these is likely for amlyopectin potatoes,
because as detailed in the responses to questions in EC-67/At.83, the amylopectin potatoes do not
have different concentrations of nitrate or glycoalkaloids compared to non-transformed potatoes. Two
other lines of argument can be considered. First, because amylopectin occurs in other crops, have
there been any observable adverse effects from grow ing these crops? Second, is there any route by
which amylopectin or nptII directly could cause a nd adverse effect? Issues related to nptII are

86 Page 8.

extensively covered in the notifica tion, and concerning direct non-target effects, the notification
appears to present a strong case that such effects are lik ely to be small or absent. In the paragraphs
below I will take each of these other points in order.
493. Other crops. Using Agricola and CAB Abst racts, I found 1213 references to waxy
maize/corn during the past 25 years. I looke d at 250 of them and found seven references 87 to the
effects of waxy corn on farm animals. Waxy corn does effect animal digestion, but the effects may be
possibly beneficial in some animals. There were no studies published on effects on other organisms.
This provides some evidence that amylopectin potatoes may have little adverse effect on mammals.
494. EC-67/At.83. Effects on nematodes. The notification reports a summary of an experiment on
cyst nematodes. The conclusion was that the transgenic potato had no effect on the nematodes tested.
The full data are not included in the notification, and there was no request for the data by the CA or
Sweden or the SCP. Without the full data, it is not possible to evaluate the scientific merit of the
conclusion.
Reply to questions forwarded by the Austrian Competent Authority.
Your question regarding experimental tests with non-target organisms, in order to
verify our statement that the clone cannot be supposed to cause any danger to other
non-modified organisms:
As we mentioned during the technical mee ting in Brussels on July 23, no such tests
have been performed. We are still convin ced that the transformed clone cannot be
supposed to cause such harm. During growing of the clone we have not been able to
note any different reactions in the interaction between the potato plants and the
insects and nematodes mentioned. The only analysis made confirms that the
transgenic clone carries the same resistan ce to cyst nematodes (Ro1,Ro4) as does the
recipient variety. It should also be mentioned that one goal for potato breeders
working with conventional methods (not gene technology) is to introduce resistance
to parasitic organisms like beetles and aphids.
88
495. EC-67/At.83. Fruit water as a possible source of risk. There appears to be little
environmental risk associated with fruit water.

87 Snow,-J.L.; Stein,-H.H.; Ku,- P.K.; Trottier,-N.L. 2004. Amino acid digestibility and nitrogen
utilization of high oil, high lysine, and waxy maize fed to growing pigs. Animal feed science and technology
113: 113-126. Akay,-V.; Jackson,-J.A.-Jr.; Harmon,-D.L. 2002. NutriDense and waxy corn hybrids: effects on
site and extent of disappearance of nutrients in sheep. J-anim-sci. 80: 1335-1343. Many factors examined,
generally few significant effects, but some differences reported. Akay,-V.; Jackson,-J.A.-Jr. 2001. Effects of
NutriDense and waxy corn hybrids on the rumen fermentation, digestibility and lactational performance of dairy
cows. J-dairy-sci. 84: 1698-1706. Many factors examined, generally few significant effects, but some
differences reported. Dado,-R.G. 1999. Nutritional benefits of specialty corn grain hybrids in dairy diets. J-
anim-sci. 77 (suppl.2): 197-207. Review. Variable effects reported. Reid,-C.A.; Hillman,-K. 1999. The effects
of retrogradation and amylose/amylopectin ratio of starches on carbohydrate fermentation and microbial
populations in the porcine colon. Anim-sci. 68 (pt.3): 503-510. Potentially positive effects on young pigs.
Kleesen,-B.; Stoof,-G.; Proll,-J.; Schmiedl,-D.; Noack,-J.; Blaut,-M. 1997. Feeding resistant starch affects fecal
and cecal microflora and short-chain fatty acids in rats. J-anim-sci. 75: 2453-2462. Potentially detrimental
effects on rats. Schroeder,-J.-W. (Jerome Wilfred); Park,-C.-S.; Marx,-George-Donald. 199-. Waxy corn as a
replacement for dent corn in diets of lactating holstein cows: final summar y. Fargo, N.D.: Animal and Range
Sciences, North Dakota State University, 14, [8] leaves: ill.
88 Page 9.

It is necessary to specify the composition of potato processing effluent, count plant
cells and provide information about the h ealth and environmental safety of this
effluent, in particular for soil bacteria flora, and the process used.
Objection raised by the CA of France.
We assume that the objection concerns the "fruit water" which is spread over fields as
a fertilizer.
The fruit water consists of water that is added to the process for washing of the
potatoes and for washing of the starch, and a small proportion of fruit juice that is not
added to the pulp.
The fruit water contains small amounts of potato skin and flesh residuals, mainly
fibres and cells, and water soluble salts of potato constituents.
Consequently, nothing is added to the e nvironment that was not present in the
potatoes, and as the water content of the fruit water is > 98%, the amounts of potato
material is very small.
It should be noted that fruit water from the st arch industry has been used as fertilizers
for more than 10 years in Sweden, without causing any problems.
The only constituent of the fruit water that could be a concern is the presence of the
nptII gene causing resistance to aminoglycosides (antibiotics). In Part I, Section D,
paragraph 9; Part 3, Section C; Annex 1 and Annex 22 of the notification, and in
attachment 8 of this document it has been shown that this cannot possibly be assumed
to cause any harmful effects by adding antib iotic resistance of any significance to the
soil bacteria population.
89
496. EC-67/At.83. Vitamin C. Amylopectin potato has higher Vitamin C content than the
untransformed clones, although the concentration re mains within the normal range of variation for
potato. Even though it is in the normal range, it is elevated, and could possibly cause a risk. While I
consider the following scenario unlikely, there is a possible risk pathway. Holometabolous insects
require enough Vitamin C to complete pupation. If pupation is limited for any insect on potato, it
could have a higher survival rate on the amylopectin varieties.
• The higher content of vitamin C indicated in the notification is confirmed. The
difference between EH92-527-1 and Prevalent is even more pronounced.
90
• In both years the content of vitamin C was significantly higher in the transformed
clone than in the non-transformed variety. This is explained in attachment xx,
vitamin C being involved in the sugar and carbohydrate synthesis of the plant. The
higher content of vitamin C is not considered to have any impact on animal health or
on the environment. It should also be ob served that the vitamin C contents measured
are within the range of natural variation of the recipient clone (table 9 of the
notification). Please note that the values reported in annex 8 of the notification and
attachment 1 of this document are expressed as mg/100 g dry matter, while the

89 Page 57.
90 Page 24.

corresponding values of table 9 of the notification are expressed as mg/100 g fresh
weight.91
Exhaustive comments on the observed significant differences in Vit.C, dry matter,
fructose, glucose and sacharose content, associated with the deliberate modification
of the starch synthesis.
Objection raised by the CA of Italy
EH92-527-1 produces less starch than the mother variety Prevalent. This is probably
because of its inability to synthesise amylose and not using the available sugars to
synthesise amylopectin instead. During tuber starch accumulation the tuber is a sink
for sugars from the photosynthesis. If those sugars are not incorporated into starch it
might result in the higher content of mono- and disaccharides in EH92-527-1.
Mutations that affect starch synthesis often result in increased levels of sugars. A
mutation with great effects on starch synthesis is where ADP-glucose
pyrophosphorylase is affected. This enzyme gives the first commited step in starch
synthesis and antisense inhibition of the ge ne results in potato tubers with decreased
dry matter, decreased starch content and increased accumulation of soluble sugars
(Müller-Röber et. al.). The modification is not the same as in the present Notification
but points in the same direction. A decreased ability to form starch results in
increased levels of soluble sugars.
When it comes to Vitamin C, another high amylopectin potato clone is currently
being investigated. This new transgenic clone has the same levels of Vitamin C as
EH92-527-1. It is to be noted that the c ontent of Vitamin C in EH92-527-1 is within
the levels normally observed in potato. A possible explanation is that the higher
Vitamin C content is an associate effect of elevated levels of mono- and disaccharides
since ascorbic acid is most probably synt hesized from simple sugars in a few
metabolic steps in plants. In a resent publication (Wheeler et. al.) the pathway
leading to Vitamin C in plants was inves tigated. In the pathway presented, glucose
and fructose are starting materials for the biosynthesis of Vitamin C. Therefore an
increase of substrate levels might very well result in the significant increase of
Vitamin C observed in the present Notification.
92
497. The transformed amylopectin potato has several transgene loci. Although most of the issues
now appear to be resolved, the discussion on molecu lar characterization implied that the plant might
have been producing an unidentified protein fro m one of these other transgene loci. If such
production had occurred, it could lead to non-target risks.
498. From the alternative perspective, it is difficult to believe that amylopectin potato would cause
a significant adverse effect on non-target species. Aside from the adverse effects of growing potato, it
is not clear what kind of change to potatoes woul d result in a significantly greater adverse effect.
Moreover, if any adverse effects do become manifest, it would be possible to withdraw potato from
the market and expect that the transgenic potato would rather quickly die out.
499. In summary, the SCP was justified in asking fo r a non-target risk assessment on 4 February
1999. In light of the data in front of it at the time, it was not justified in concluding on 12 July 1999

91 Page 28.
92 Page 62.

that amylopectin potato would have no adverse effect s on non-target organisms. The scientific data
provide no evidence that there is an adverse effect , but the data are not strong. Hence the scientific
data and scientific arguments can divide on whether there is a necessity for additional non-target risk
assessment. On the one hand, some could argue credib ly that adverse effects on non-target organisms
are unlikely, and no additional assessment is needed. Others could argue credibly that adverse effects
are possible, requiring data to address the few loose ends identified above. In no case, however, could
it be credibly argued that adverse non-target effects are likely to or in fact do occur, and therefore an
extensive non-target risk assessment is NOT necessary.
500. The SCP requested information from the notifie r on 4 February 1999. The notifier provided
the full notification on 4 January 1999. The 31 days th at elapsed is a reasonable, if not rapid response
time.
Dr. Healy
Note: the question specifies the Scientific Committee for Food, however there is no mention of this
committee in any of the documents, the numbered a ttachments all refer to information requested by
the Scientific Committee on Plants (SCP). I have answ ered this question in relation to the information
requested by the SCP.

501. Response: Following receipt of the notification from Amylogene seeking approval for
Amylogene starch potato clone EH92-527-1 in A ugust 1996, the lead CA (Sweden) expressed a
favourable opinion in May 1998. The dossier w as then sent to the European Commission and
distributed to other Member States for comment . A number of comments and requests for further
information were received from various Member St ates. In September 1998 the EC requested the
SCP to consider whether there was any reason to be lieve that the placing on the market of the starch
potato clone EH92-527-1 would be likely to cause any adverse effects on human health and the
environment.
502. The dossier and additional information were supp lied to the SCP in January 1999. Between
February 1999 and November 2001, the SCP made ni ne further requests to Amylogene for additional
information or clarification of information.
503. Background: Potato clone EH92-527-1 contains two novel genes, an antisense Granule
Bound Starch Synthase gene (gbss), and a marker gene, the nptII gene. The nptII gene, which is
widespread in nature, is widely used in GM plan ts and is well characterised. The gbss gene was
isolated from the potato and was reintroduced into the potato in the antisense orientation to silence the
endogenous gbss gene, which is involved in the synth esis of amylose. This results in the production
of starch with high amylopectin content. No protein is produced from this introduced gene.
504. The potato clone EH92-527-1 was generated pr imarily to produce starch for industrial
purposes. The by-products of this process, potato pulp, potato juice and potato water, were intended to
be used for animal feed and irrigation purposes. No products derived from potato clone EH92-527-1
were intended for use as human food.

505. Molecular characterisation:
4 February 1999 (EC-67/At. 84): the SCP requested further evidence that the plasmid 'backbone'
DNA was not incorporated into the genomic DNA:

More convincing evidence is required that speci fic elements of "backbone" DNA are or are not
incorporated
(A) The dossier lacks a detailed map or tabl e describing the DNA backbone outside T-DNA
borders. One or other should be provided. Or igins of all open reading frames and control
elements should be given and a risk assessment provided for each. The notifiers should clarify
the antibiotic specificity of the nptII gene incorporated into the backbone of the binary vector.
(B) There are some confusing and inconclusi ve PCR analyses to demonstrate absence of
vector sequences flanking the border regions. Appr opriate controls are lacking (i.e. positive
PCR's are carried out on plasmid DNAs instead of control genomic DNAs or mixtures of
genomic and plasmid DNA). The use of overlapping probes in Southern analysis is preferable.
(C) Southern blots have been used to determin e integration of genes/ gene fragments outside
left and right T-DNA borders. A cocktail of probes has been used. However, there is no positive
control to demonstrate that all elements of the probe are working.
(D) Also related to Southern analysis, only 2 probes have been used to determine copy
number. No probes have been used covering the nos gene for example.

506. Response: The data in the original dossier concerning the molecular characterisation of
potato clone EH92-527-1 included Southern blots of genomic DNA from both the transgenic potato
and its parental line, using insert and backbone spec ific probes. Positive and negative controls were
also performed. PCR analysis of the junction re gions between the insert and the genomic region
indicated that no large fragments of introduced DN A were present at these junctions. However, no
sequence data were provided.
507. Southern analysis was originally conducted using a cocktail of backbone specific probes,
although this does not conclusively show the absen ce of plasmid backbone sequence. Evidence was
provided that the company intended to supply conclusive data showing no plasmid backbone
sequence is present is potato clone EH92-527-1 and DNA sequence of the junction regions (EC-
67/At. 83). However, it is not clear from the docum ents provided (Exhibit EC-67) whether this data
was actually provided to the EC or the SCP. This in formation, together with the data in the dossier
provided to the SCP would have made the requests at A), B) and C) unnecessary.
508. A detailed description of the DNA backbone outsi de of the T-DNA border is not warranted
(part A) if sufficient evidence is provided to show that no backbone DNA is present in the DNA of the
GM potato.
509. The optimal number of probes to be used in S outhern hybridisation is determined on a case-
by-case basis (part D). Potato clone EH92-527-1 contains two additional genes the gbss gene and the
nptII gene, and therefore as a minimum it would be r easonable to use probes to detect each of these
genes. The company used probes specific for the gbss gene and its promoter region. Due to the
presence of an endogenous gbss gene, this experimental approach was less than optimal, however, the
use of the non-transgenic potato DNA as a negative c ontrol allowed identification of the endogenous
gene. Although the initial selection procedures confirm the presence of the nptII gene, probes
detecting the presence and copy number of this gene in potato clone EH92-527-1 would be required.

510. Toxicity and animal feeding trials
4 February 1999 (EC-67/At. 84): the SCP requested information on the use of potato pulp for animal
feeding in different MS, including information on targ et animals. The SCP also requested justification
for the absence of a feeding trial using the GM potato:

The notifier should provide information on possible application of pulp for animal feeding in
different EU Member States. Target animals etc.

The notifier should justify the absence of an y toxicological data involving relevant animal
feeding trials.

511. Response: Potato pulp has a long history of safe use as stock feed. The pulp of the non-
transgenic parental line had been used in Sweden as cattle feed since 1988 without known harmful or
negative effects. Evidence was provide that the nutritional composition of tubers from potato clone
EH92-527-1 was similar to that of the parental line and within the normal reference ranges for
commercial potatoes, except for those intended di fferences related to starch composition. The
Swedish Board of Agriculture notified its approval of the transgenic potato for placing on the market
in May 1998.
512. The starch present in the pulp from EH92-527-1 differs from common potato starch
(composed of amylose and amylopectin) only by lack of the amylose component. The company
provided data and estimates of consumption rates in cattle of potato pulp. As amylose usually makes
up 20-25% of the total starch in potatoes, if pul p from potato clone EH92- 527-1 were to be used,
cattle would consume no amylose and more amylopectin per day.
513. Other agricultural crops, such as waxy maize, contain starch with similar levels of
amylopectin to potato clone EH92-527-1. Rice, barley and sorghum also contain high levels of
amylopectin. Furthermore, the potato pulp used as stockfeed is a by-product of the starch industry and
would therefore be unlikely to contain high levels of starch.
514. Given the compositional analyses of the transgenic potato, the similarity in starch
composition with other plant derived animal feeds, and the absence of starch remaining in the potato
pulp, feeding studies using potato clone EH92-527-1 in various livestock species are not necessary to
support a valid safety assessment.
9 March 2000 (EC-67/At. 99): in light of the presen ce of an ORF in potato clone EH92-527-1 (ORF4,
discussed below), the SCP requested an animal feeding study:
Evidence should be provided regarding the safety of the material for animal feeding since part
of the by-products will be used for animal feedin g. This is of relevance given the presence of a
number of ORFs which show significant homolo gies to known coding regions. The safety may
be assessed by performance of a 90 day feeding tr ial with laboratory animals administered diets
with different levels of potato pulp (concentrated).
515. Response: Further molecular characterization data identified a putative open reading frame
(ORF, designated as ORF4) in the transgenic potato. Although transcription was detected, there is no
evidence that ORF4 is translated into a protein. Potato tubers are predominantly composed of
carbohydrate with only minimal amounts of protein. In this case, the transgenic potato has been
shown to be compositionally equiva lent to conventional varieties, except for the intended difference

relating to the starch component. Feeding studies using target livestock species will not add
substantially to the safety assessment because low dietary exposures to potato proteins would not
yield meaningful results that could be validly interpreted.
516. Protein analysis
4 February 1999 (EC-67/At. 84): the SCP request ed information on the expression of any novel
proteins in potato clone EH92-527-1:

In addition to the requirement for conclusive Southerns for components of backbone DNA the
Committee would prefer to see Northern/Western analysis where appropriate. For example, the
expression of the nptII gene and other open reading frames incorporated into the GM plant.

517. Response: Western analysis showing the lack of a gbss gene product in potato clone
EH92-527-1 was provided. These results demonstrate the effectiveness of the genetic modification.
Given that the introduced antisense gbss gene has a silencing function, the results of Northern
analyses would add little to the assessment of safet y of the transgenic potato. Messenger RNA from
the endogenous gbss gene would normally be detect ed and no additional safety concerns would be
raised by the presence of the antisense gbss transcripts.
518. However, the dossier contained no data on the expression of the nptII gene. As the only novel
gene expected to be expressed in the transgenic potato, data and information on the expression of
nptII would be required. Rather than Northern analysis, antibody based techniques (such as ELISA)
would be sufficient to determine the levels of APH(3')II expressed in the tubers.
519. Several other ORFs are present in the inserted DNA. In the first instance, bioinformatic
analysis of any hypothetical polypeptides would be an appropriate means of assessing the significance
of these in the potato. Depending on these resu lts and the presence of any putative regulatory
elements, Northern analysis may be justified. This analysis would be particularly important where
significant homology with known protein toxins or allergens was identified, or where information
from other parts of the safety assessment indicated a potential safety issue.
520. ORF4: Over the period from June 1999 to November 2001, the SCP required further
information on an identified ORF in the inserted DNA, designated ORF4. ORF4 lies between the 3'
end of the nptII gene and the nospA element (the polyadenlation signal) and is not in the same reading
frame as nptII. ORF4 was identified as having some homology to the bleomycin resistance protein of
Tn5 (E. coli) and the ornithine cyclodeaminase protein (from Agrobacterium tumefaciens).
3 June 1999 (EC-67/At. 85): the SCP requested the following information from Amylogene:
ORF analysis indicates homologies within ORF4 to the bleomycin resistance protein of Tn5.
Bleomycin is a chemotherapeutic used in cancer treatment. The SCP wishes to know the
significance of the presence of this ORF with re spect to the expression of a protein and to the
possibilities of transfer of a functional bleomyci n resistance gene to non-target organisms. This
could be determined by Northern blot analysis of potato leaves and tubers.

521. Response: The safety assessment includes an evaluation of the significance of unintended
translation products arising from the genetic modification on human health and safety. Evidence
would be required to ascertain whether expression products could arise from ORF4, and including an
evaluation of the potential toxicity and allergenicity of these products. Northern blot analysis would

normally be expected to detect transcription of functional genes, but due to a lack of sensitivity, may
not detect low abundance messages.
522. The exquisitely sensitive technique of rever se-transcriptase PCR (RT-PCR) was performed
and transcription across the nptII gene into ORF4 was detected. However, this result does not
demonstrate that translation of ORF4 would occu r. As ORF4 lies between the nptII gene and its
polyadenylation signal, a transcript corresponding to this region of DNA may be expected.
Furthermore, the presence of a stop codon immediately following the nptII gene strongly suggests that
translation stops at the 3' terminus of this gene. In addition, ORF4 is not in the same codon reading
frame as the nptII gene and would therefore require additional regulatory signals for translation.
523. Theoretical evaluation of the safety of any translation product from ORF4 is required in the
assessment. Only the first 50 potential amino aci ds of ORF4 are homologous to the bleomycin
resistance gene (total of 126 amino acids). This peptide has been shown to have no bleomycin
resistance activity. Therefore, in the unlikely event of transfer of this putative coding region to micro-
organisms, bleomycin resistance would not be transferred.
524. Additional bioinformatic analysis would dete rmine if ORF4 has any homology with any
known protein toxins or allergens. However, given that there are no indications that ORF4 produces a
protein and that exposure to any proteins from potato clone EH92-527-1 by animals is likely to be
extremely low, the transcription of ORF4 does not raise any safety concerns.
525. Therefore, the additional information requested by the SCP on the potential expression of
ORF4 would contribute to the validity of the safety assessment. It should be noted that in seeking
information from the company, the SCP could have anticipated the need for additional molecular and
expression analysis data concerning ORF4. For example, sequence analysis of ORF4 to detect any
homology with known protein toxins or allergens was indicated on identification of ORF4.
9 March 2000 (EC-67/At. 99): the SCP requested Amyl ogene to conduct bioinformatic analysis of the
amino acid sequence of ORF4 to provide information on the possible allergenicity of any polypeptide
that might be produced:

The SCP accepts the notifier's evidence that no functional bleomycin resistance protein is
produced from ORF4. However, the SCP asks the company to provide information on the
possible allergenicity of any polypeptide that co uld be produced. The SCP accepts that this can
be completed by comparing sequence homologies using relevant databases.

526. Response: It is noted above that these data are indicated in this case to assess the safety of
potato clone EH92-527-1 and therefore in my opinion were required as part of the dossier.
20 July 2000 (EC-67/At. 100): the SCP requested Amyl ogene to repeat this analysis to search for
similarities within ORF4 without any reference to the word "allerg" in the query:
Swiss prot databases have been queried for entrie s with sequence ID's combined with any word
beginning with "allerg" in any field. For completeness the SCP asks the notifier for a full search
for, and list of, similarities within ORF4 without reference to the word "allerg" in the query.

527. Response: This second search is unlikely to add any new information to the assessment of
ORF4 for potential allergenicity. It may, however, identify sequence similarity with known toxins in

the database, as it appears that this specific information had not been supplied to the SCP at this stage.
This analysis would be consistent with a thorough safety assessment.
20 July 2000 (EC-67/At. 100): Amylogene was requested to produce antibodies to the ORF4
polypeptide so as to be able to test for expression in potato clone EH92-527-1.

The SCP thanks the notifier for their efforts in attempting to prepare recombinant polypeptides
using ORF4 and expression vectors. The notifier is requested to attempt antibody production
using synthetic polypeptides raised to immunogenic sequences within ORF 4. This should
include sequences in ORF4 similar to MP12 AMBAR. Several peptides should be prepared
from sequences spanning components of ORF4 to improve the chance of success. Antibodies
could be raised in mice, in addition to rabbits, to accelerate the process of risk assessment.

528. Response: Detailed bioinformatic analyses did not indicate homology of the putative ORF4
amino acid sequence with any known protein toxins or allergens. Furthermore, as stated above, there
is no evidence that ORF4 is translated. Based on the totality of the information on potato clone EH92-
527-1 and its intended uses, sufficient information is available to support the conclusions of the safety
assessment. Further analysis including Western bl otting requiring ORF4 specific antibodies is not
justified.
Between 20 July 2000 and 8 November 2001, the SCP made a number of requests to Amylogene,
concerning the Western analysis of potato clone EH92-527-1 in relation to expression of ORF4. These
questions are listed below:

1 February 2001 (EC-67 Attachment 102)

It is necessary to see an alignment of the amino acid sequences used to generate the synthetic
peptides and the original nucleotide sequence.

In Western blots, figure 1(b), the immunoresponse of the GM leaf protein in lane 12 is very
different from that of the parent line. In lane 12 two major bands are missing ca 40 and 55kDa,
yet a band appears to occur between the 40 and 55kDa proteins (ca52kDa?). This band appears
to be present in lanes 4 and 5 but not in lanes 7, 8, 9, and 11. This implies that expression of
ORF4 results in the formation of a polypeptide much greater than 17.5 kDa. lane 12 implies that
this polypeptide and ca 52kDa, resulting from expression of ORF4 in E. coli is in fact presenting
the GM line. The interpretation of these data by the notifier is requested.

Proteins in lane 12 do not appear well resolv ed compared with lanes containing non GM-
derived leaf protein. There is a need to repeat the blots with adequate replication and with
coumassie stained gels shown in parallel. The notifiers should consider Western blots with
tubers as well as leaves.

Western blot the purified ORF4 polypeptide from E. coli not a crude E. coli extract.

14 March 2001 (EC-67 Attachment 104)

Statements that recombinant ORF4 is a 17.5 kDa protein whereas in the GM plant the
polypeptide would be 14.4 kDa. Fig 5 indicates that the recombinant protein is >25 kDa?

In Fig 4c a cross reacting band appears at 48 kDa in all lanes with leaf proteins from the GM
line (lanes 3-9, lane 12) but not in the non GM parent (lane 11). This band cannot represent E.
coli protein as its intensity does not changes with dilution of the partially purified ORF4. This
band was commented upon in an earlier question to the notifier (see Fig 1). We also note that
the Western blot pattern for Fig 1 is significantly different from Fig 4 (with the same samples?)

With respect to dilutions of ORF4 used to "spike" potato protein samples, there appears to be
very distinct differences in sensitivities for gels in Figs 4 b and d in particular then the presence
of any ORF4 in tubers may be missed?

The coomassie stained gel shown in Fig 4e (which represents the same samples in each lane
shown in Fig 4c) indicates that the polypeptide profile in lane 11 (the non GM parent) is very
different from the profiles for the GM line. Does this indicate a lack of substantial equivalence?

The SCP would also like to discuss the numbers and nature of replicate leaf and tuber samples
chosen and the nature of any additional work th at might be needed to formulate a final risk
assessment.

3 April 2001 (EC-67 Attachment 105)

The notifier should provide details on the contruct used to express ORF4 in E. coli

Since there is still a question over the ca 48 kDa protein it will be important for the notifier to
prove their hypothesis that the band is non spec ific and represents an endogenous, naturally
occurring potato protein. This will mean ca rrying out Western blots with samples from
equivalent leaf positions using plants of equi valent age and physiological status. Leaves should
be used from several non GM and GM plants and experiments for replication purposes. The
notifier could also include comparisons with other antisense GBSS potato GM lines that
company might hold to emphasise their argument . Replicated tuber Western blots would also
be very beneficial. It will be important that the sensitivities of the various Westerns are as
similar as possible so that any relevant immuno logical cross reactions are not missed. Spiking
some lanes with ORF4 protein, as done previously, appears to be a good way to approach this.
In addition to the Western blots, parallel SDS- PAGE and coomassie stained gels of leaves and
tubers will also allow the SCP to determine eq uivalence of the samples selected in terms of
overall polypeptide profile.

8 November 2001 (EC-67 Attachment 106)

The SCP requires that all data proceed in response to questions to notifiers to be of a standard
acceptable for publication in a well-regarded peer reviewed journal.

The quality of Western blots provided to date and using antibodies raised against ORF
sequence is not considered adequate. More specifically:

The sensitivity of the detection system appears highly variable from blot to blot for known
amounts of ORF4 protein. In SDS-PAGE and co omassie stained gels control ORF protein is
visible in come cases but absent in others. This raised questions on the overall reliability of data
presented.

It may be pertinent for the notifier to consider sub-contracting the Western blotting to a third
party.

The notifier should seriously consider additi onal Western blots using antibodies raised
specifically against the nptII protein. This will add weight to arguments that the polycistronic
RNA associated with ORF4 does not give rise to a fusion protein (molecular mass greater than
that of nptII protein alone).

529. Response: The totality of the data and analysis of ORF4 already supplied to the SCP
supports the validity of the safety assessment without the need for Western blot analysis of ORF4.
530. APH(3')II (nptII gene product)
4 February 1999 (EC-67 Attachment 84 Question 6): th e SCP requested that the levels of APH(3')II
protein be provided for tubers and plant tops:

The concentrations of NPTII protein should be provided for tubers and plant tops (haulms).

531. Response: A number of countries, including Canada and Australia would usually require this
information for the safety assessment of a GM plant intended for use as human food. However, the
guidance document for preparing dossiers (SCP, 1999), states that this information may be required if
a potential risk is identified.
532. The nptII gene was used as a selectable marker gene in development of the potato clone
EH92-527-1 and is expressed in the transgenic potato. It is generally accepted that information on
expression levels of novel proteins in GM plants are required for the safety assessment.
533. The nptII gene has been used often as a marker gene in transgenic plants. From previous
evaluations it has been established that the nptII gene and resistance to kanamycin and neomycin are
ubiquitous in nature and that neither the gene nor its protein product pose a risk to human, animal or
environmental health when used in GM plants. It is therefore unlikely that information on the
expression levels of nptII in potato clone EH92-527-1 would adversely influence the safety
assessment. However, expression levels would be of in terest to determine that patterns of expression
in the transgenic potato are as expected. Therefore this request is warranted to ensure the safety
assessment is comprehensive and consistent with other regulatory bodies internationally.
3 June 1999 (EC-67/at. 85): In vitro data on the su rvival of the APH(3')II protein from leaf tissue in
the presence of rumen micro-organisms (and of ORF4, if expressed) was requested by the SCP:

In vitro data on the survival of nptII protein, preferably supplied as leaf tissue in the presence of
rumen micro-organisms should be provided. Should ORF4 be expressed and produce a protein
then the extent of its digestion in the presence of rumen micro-organisms should also be
determined.

534. Response: There is concern that horizontal tran sfer of antibiotic resistance genes from
transgenic plants to gut micro-organisms may cont ribute to increased resistance against clinically
important antibiotics. The potential impact this may have on human health is assessed on a case-by-
case basis.

535. However, as mentioned previously, the nptII gene is already ubiquitous in nature and has
been considered by the World Health Organization to pose no additional risk to human or
environmental health when used in GM plants (WHO, 1993). Although ORF4 has some homology to
a bleomycin resistance gene, is not functional a nd therefore would not confer resistance and would
pose no additional risk if it were to be transferred to micro-organisms in the gut.
536. Effects on non-target organisms
4 February 1999 (EC-67/At. 84): the SCP queried why no data were provided on the safety of
modified crops to non-target organisms:
No data are provided on the safety of modifi ed crops to non-target organisms. No risk
assessment has been made.

537. Response: The nature of this transgenic crop is not such that effects on target organisms
would be anticipated. Compositional analysis did not identify any changes in the nutrient or
antinutritional profile of the GM potatoes that w ould suggest concern for non-target organisms.
Agricultural practises for this crop are identical to conventional starch potatoes. Amylogene did not
observe any unusual interaction between the transgen ic potato plants non-target organisms, such as
insects, during field trials of this crop.
538. Given this information, further data on the effects of the transgenic potato on non-target
organisms are not warranted to support a valid safety assessment.
539. Bleomycin resistance
3 June 1999 (EC-67/At. 85): the SCP requested cl arification on the potential of ORF4 to provide
resistance to bleomycin and the possibility of transfer of a functional bleomycin resistance gene to
non-target organisms:
 ORF analysis indicates homologies within ORF4 to the bleomycin resistance protein of Tn5.
Bleomycin is a chemotherapeutic used in cancer treatment. The SCP wishes to know the
significance of the presence of this ORF… with resp ect to possibilities of transfer of a functional
bleomycin resistance gene to non-target organism.

540. Response: The guidance document for the preparation of dossiers (SCP, 1999) and the
Codex Guideline for the Conduct of Food Safety Assessments of Foods Derived from Recombinant
DNA Plants (CAC, 2003) recommend that the safety assessment of GM plants should take into
account the potential impact of horizontal DNA tran sfer between the plant or plant components and
micro-organisms.
541. Therefore, it important to determine whether the potential ORF4 polypeptide would confer
bleomycin resistance and what impact this would have on human health and the environment if a
functional bleomycin gene were to be transferred to other organisms, for example micro-organisms.
542. Substantial equivalence
4 February 1999 ( EC-67/Att. 84): the SCP requested additional statistical analysis of the
compositional data:

Evidence of substantial equivalence: data from 1996 and 1997 should be treated for statistical
purposes as a single data set and, ideally, strengthened by including data from the 1998 season.
To fully justify co-variant analysis and conclusi ons derived from it, the interdependence of yield
and composition should be addressed for two to three growing Seasons
543. Response: The SCP guidance document for the preparation of dossiers (SCP, 1999) indicates
that compositional analyses should be based on samples from at least two growing seasons, with the
growing sites being diverse in number and locati on, and accompanied by appropriate statistical
treatment. The information provided by Amyl ogene indicates that the samples analysed for
composition purposes were derived from two growing seasons, namely 1996 and 1997. In each year,
three trials sites were used.
544. The SCP guidance document also indicates th at that the specific analyses should be
appropriate to the crop. In the case of potatoes, fi ve constituents have been identified as important for
compositional comparison, namely dry matter, sugars (especially reduc ing sugars), protein, vitamin C
and glycoalkaloids (OECD, 2002). However, several other key c onstituents were also identified
including starch, fat, fibre, minerals, protease inhibitors and lectins. Amylogene submitted
information on the five constituents recommende d by the OECD Taskforce on Novel Foods and
Feeds as well as the majority of the other constituents, the notable exceptions being protease
inhibitors and lectins.
545. The results from the compositional analyses indicat e that the majority of the constituents are
produced in similar levels in the GM and non GM lines. The levels of three constituents, nitrate,
glycoalkaloids and fibre, appear to differ between the two potato varieties in one of the growing
seasons only and thus the differences do not appear to be specific to the variety of potato. The
analyses also identified several cons tituents that show statistically significant differences between the
GM and non GM potatoes, including dry matter (may be related to changes in yield), vitamin C and
sugars. These differences are consistently observe d over the two growing seasons examined and thus
appear to be variety specific.
546. To determine if the differences observed in the le vels of vitamin C and sugars are likely to be
biologically significant, the levels observed in both potato varieties were compared to the levels of the
constituents present in the non GM potatoes grown under different environmental conditions. This
analysis appears to have been presented for the in formation from the 1996 trial only. In all cases the
levels of the three sugars tested and vitamin C fa ll within ranges present in other varieties of potato
that are readily available. It is therefore unlikel y that the detected differences in composition pose a
threat to human health and safety. Further information from another growing season (ie 1998) is
therefore not necessary to ensure the validity of the safety assessment.
547. Amylogene undertook a co-variant analysis on pa rt of the data set, apparently to analyse
possible effects of the reduced yield on levels of particular constituents. Given this approach, it is
reasonable to analyse the data from the two gr owing seasons both as single data sets as well as a
combined data set. However, this analysis is not necessary to ensure the validity of the safety
assessment for the reasons outlined above.

548. Other Issues
February1999 EC-67/Att 84 4: the SCP requested information about the fractionated potato products:

Concerning the overall use of the GM plant it would be beneficial to have a fractionation
scheme showing how by-products arise and the conditions used for extraction. Relevant analysis
of each by-product entering the food chain is required.

549. General information on the fractionation of the potato was provided in the original
application. The genetically modified potato will be used primarily for starch production. By-
products of starch production are pulp (solid remains of the potato after the st arch has been extracted)
potato juice (liquid from the potatoes) and potato water (water used to wash the potatoes). The juice
and water are used for irrigation and the pulp is used for cattle feed. The potato and its products are
not intended to be used for human food, nor is it intended that the whole tubers be used for animal
feed.
550. The safety assessment, addressing the molecular characterisation, properties of any newly
introduced substances and compositional analysis, h as not identified any health and safety issues
arising from the genetic modification. The only co mpositional difference of significance is the type
of the starch, with the genetically modified potat o having high levels of amylopectin. However, a
range of conventional crops produce the amylopectin ty pe of starch (eg maize) and amylopectin type
starch is also chemically isolated from potatoes. Ther efore, the safety of this type of starch is well
established (see also response to questions on Toxicity and animal feeding trials).
551. The information requested on fractionation of by-products and analysis of the by-products
entering the food chain was not necessary to support a valid safety assessment.
552. Overall Conclusion to Question 25: The SCP was not initially provided with adequate
information to enable a valid safety assessment to be conducted. However, much of the information
requested by the SCP would not have had an impa ct on the conclusions of the safety assessment. It
also should be noted that in seeking information from Amylogene, the SCP could have anticipated
much of the data required and requested this from the company in a more timely and coordinated
manner.
553. In conclusion, the totality of the information provided to the SCP up to 20 July 2000 was
sufficient to enable a valid safet y assessment to be conducted. The information requested after this
date (and much of the information requested prior to this date) is unlikely to influence the conclusions
of the safety assessment.
Dr. Snape
554. The comments below relate to Question 25 con cerning only the molecular characterisation of
exhibit EC-067. These were examined relative to the EFSA-Q-2003-005 guidance document
produced by the Scientific Panel on Genetically Modified Organisms.
555. Exhibit EC-067: Potato event EH92-527-1 was produced, using plasmid pHoxwG, by means
of Agrobacterium-mediated gene transfer . The T-DNA of the plasmid pHoxwG used for
transformation carries two expression units: one contai ns the potato gbss gene in antisense orientation
and one contains the nptII antibiotic resistance gene. Potato event EH92-527-1 contains one
functional transgenic locus containing one copy of the gbss antisense gene. Information relative to the
molecular characterisation of EH92-527-1 has been found primarily in EC-067 attachments 14, 18,

20, 21, 36, 37, 39, 41, 42, 43, 44, 60, 82, 107, 108, 111 and 113. A series of comments and questions
are numbered below. This is followed by a conclusion proposing answers to Question 25:
556. EC067 – attachment 021: (1) The southern blot analysis seems to suggest the presence of
three copies of the gbss promoter (figure 3) and of the gbss gene (figure 8) in wild-type Prevalent
potatoes. This contrasts with the results obtained in figure 6 suggesting that only one copy of the gbss
gene is present in wild-type Prevalent (one hybridization signal/band in lane D).
557. EC067 – attachment 021: (2) No molecular information is provided on the nptII gene
integrated into EH92-527-1. This reviewer would suggest to re-probe membranes shown in figure 3,6
and 8 with an nptII probe.
558. EC067 – attachment 036: (3) PCR analysis of the T-DNA integrated into EH92-527-1
suggests the loss of parts of the left T-DNA seque nce in EH92-527-1. More information would be
required to characterise the extent of the T-DNA deletion at the left border.
559. EC067 – attachment 037: Shows the absence of transgenic sequence in the chloroplastic
genome of EH92-527-1 using Southern blot analysis and "labelled fragments" of the pHoxwG vector
used as probes.
560. EC067 – attachment 039: (4) Shows the absence of vector backbone sequences in EH92-527-
1 using a "probe cocktail" from a ~8000 nt BglII fra gment described in attachment 60. The use of
smaller, well defined probes from the backbone (nptIII gene etc…) would have provided more
accurate information.
561. EC067 – attachment 041 to 44: Provides maps and the entire sequence of the pHoxwG vector
used for potato transformation.
562. EC067 – attachment 060: It is mentioned that new experiments will be undertaken to detect
backbone sequences and to sequence genomic flanking re gions. (5) This reviewer disagree with the
following sentence on page 8 of the PDF file "Annex 21 does not give a clear cut case when it comes
to unwanted vector sequences in other parts of the genome of EH92-527-1 than associated with the
inserted integrated DNA". The absence of hybridisa tion signal in the Southern blot hybridisation
conducted in Annexe 21 (i.e. attachment 39) is enough to prove that no backbone sequence is
integrated anywhere in the genome of EH92-527-1 (w ith the limitations associated with the efficacy
of using a probe cocktail).
563. EC067 – attachment 86 and 113: (6) Suggests that the sequencing of genomic flanking
regions has been performed. This reviewer was unable to find this information among the attachments
presented.
564. EC067 – attachment 111: (7) This reviewer could not find the molecular analysis showing
that EH92-527-1 exhibited "genotypic stability" as claimed page 7. Comparative Southern blot
analysis of primary transformants and plants produced through vegetative multiplication is required to
assess the structural stability of a transgenic locus.
565. Conclusion – response to Question 25: Since the first submission in 1996, the following
molecular information about EH92-527-1 has been established:
• EH92-527-1 contains one functional locus c ontaining a single copy of the gbss gene
in antisense orientation.

• Some deletion has occurred in the left T-DNA border region.
• Transgenic sequences outside the T-DNA are unlikely to have been integrated into
EH92-527-1.
• Transgenic sequences from pHoxwG are unlikel y to have been integrated into the
chloroplastic genome.
566. I would suggest that more information is still required about:
• The sequence of the transgenic locus especially around the deleted left T-DNA
region. The proposed architecture of the tr ansgenic locus currently relies only on
PCR-based studies.
• The sequence of the flanking plant genomic regions.
• Additional Southern analysis (using sm all well defined probes) to confirm the
absence of backbone sequences into the genome.
• The molecular analysis of plants propa gated vegetatively to demonstrate the
structural stability of the transgenic lo cus across multiplication cycles. The stability
of the altered starch composition trait can be used to estimate the genotypic stability
of the gbss expression unit as only one transgene copy is present in EH92-527-1.
567. Additional information may have been submitted to the EC by the company. Experts should
examine if this information answers some of the requests formulated above.
Monsanto Roundup Ready oilseed rape (GT73)
C/NL/98/11 (EC chronology 70)

Question 26: Given the information before the Pane l, including the notification (EC-70/At.1-3),
was the information regarding molecular charac terization of this product requested by the
Dutch CA and by the UK (EC-70/At.4, 7, and 8) necessary to ensure that conclusions of the
safety assessment were valid?
Question 27: Given the information before the Panel, including the notification, was the
information regarding feed safety aspects of this product requested by the Netherlands (EC-
70/At.8 and 13) necessary to ensure that conclusions of the safety assessment were valid?
Dr. Nutti
568. Based on the information provided by the applicant in EC-070/At.8 and 13, and the analysis
done by the EC SCP and according to the Code x Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants (Page 4, paragraph 53), some foods
may require additional testing; regarding animal feeding studies, extra studies may be warranted if
changes in the bioavailability of nutrients are exp ected or if the composition is not comparable to
conventional foods. Although the prod uct was concluded to be substa ntially equivalent, the notifier
carried out animal feeding studies with rainbow trout , quail, broilers and lamb. In all these studies, no
differences were detected among the animals fed conventional and GM oilseed rape based feeds.
Therefore, it is concluded that the studies request ed by the Netherlands we re not necessary to ensure
the product's safety assessment.

Question 28: Given the information before the Pa nel, including the notification and the SNIF
(EC-70/At.49-53), was the additional information provided in Monsanto's submission to the
Commission (EC-70/At.84-97) necessary to ensure that conclusions of the safety assessment
were valid?
Dr. Nutti
569. Based on the information provided by the applicant in EC-070/At.1 to 3, and the analysis
done by the EC GMO Panel, where it was stated th at GT73 oilseed rape is as safe as conventional
oilseed rape for humans and animals, and in the context of its proposed use, I concluded that no
additional experimental studies were necessary, I un derstand that the additional information provided
by the notifier was not necessary to ensure that conclusions of the safety assessment were valid.
Dr. Andow
Necessity of additional information on monitoring plan:
570. In this answer I will address only the monitori ng issue. Here the question becomes "is a
monitoring plan necessary to ensure that conclusions of the safety assessment were valid?" The scope
of the notification is for import and use of GT73, excluding the cultivation of varieties derived from
GT73.
571. The monitoring issues are summarized by the notifier in EC-70/At.85.
6. Monitoring
AUSTRIA: The scenarios set out at point 2.3 and the properties of rapeseed oil also
facilitate the appearance of the product in various conventional rapeseed oil
cultivation areas or adjacent locations, even though probably only as a small
proportion. Nevertheless, this fact must be accordingly taken into account by
consideration in a monitoring programme in accordance with appendix VII f the
guideline 2001/18/EG. However, this is not allowed for on the part of the applicant
(cf. test report by the competent Dutch authorities).
DENMARK: The monitoring plan should incl ude observation on dispersal and gene
transfer to oilseed rape and wild relatives.
ITALY: The company shall not only indicate the main places of entrances in each
country but also the storage location and the wa y of the transport; this is due to the
need of guarantee an efficient monitoring plan. In any case the processing plants
should be nearby the places of entrance and every measure to avoid the dispersal of
grains into the environment must be ensured.
SPAIN: The current scope of this application fo r consent to place GT73 oilseed rape
on the market includes both import and processing in the European Union, excluding
cultivation. Nevertheless, assuming that an unintentional release could be raised, the
Spanish Competent Authority considers that a post-market monitoring plan is needed
to consider this aspect. The monitoring plan should include sampling in order to
carry out studies on transgene expression on wild relatives, conventional oil seed
rape crops, volunteers growing across the transit way during import and processing.

UK: The UK requests to monitor for the presence of feral GT 73 oilseed rape at the
points of import and processing, and betw een those locations and proposes to
establish an action plan should feral GT73 be identified … The proposal for post-
market monitoring is not adequate. As outlined above, the environmental risk
assessment assumes that seed will not be spilled, so the post market monitoring plan
should test this assumption. The UK requests that the applicant gives further
consideration to the post-market monito ring plan, which should include (i)
monitoring for the presence of feral GT73 oilseed rape at the points of import and
processing, and between those locations; and, (ii) proposals for action should feral
GT73 be identified.
93
572. The full context for understanding Austria's concerns is provided in EC-70/At.77, 25 March
2003, Objections raised by Austria, and is reproduced below. Austria is concerned about accidental
release during transport or processing and subsequent invasion of other agricultural systems.
• 2.3 Ecology: Importing the product for processing and use for food and animal feed is
no guarantee that GT73 will not be released into the environment. Experiences and
checks in Austria during the last few growing seasons have shown that genetically
modified seeds can come into circulation as a result of accidental or technically
unavoidable contamination (e.g. in the transport or processing stage). In the case of
oilseed rape, with its specific characteristics (ability to survive, potential for dispersal
and outcrossing), there is a possibility of it spreading into agricultural ecosystems.
573. This could therefore affect the possibilities fo r coexistence between different agricultural
production systems with and without GMOs. The pr oduct cannot be approved until there is EU-wide
clarification of the possibilities for coexistence and a legally binding framework (e.g. specifying
threshold values) has been defined.
574. The full context for understanding Italy's concer ns is provided in EC-70/At.71, 24 March
2003, Objections raised by Italy. Italy requests monitoring to provide a means for managing risks
associated with AMPA.
2) As for the previous notification C/ES/00/01, there is an extensive discussion
of possible effects on animal and human health (due to antinutrictional factors), but
there is no data on the possible content of gliphosate and/or metabolites (AMPA). In
this regard, we note that event GT73 contains a gene specific for the degradation of
gliphosate into AMPA, but see no discussion of the possible consequences.
94
575. The Spanish CA expands on their concerns in EC-70/At.75, 25 March 2003, Request for
additional data from Spain. Spain is concerne d about accidental releases during transport or
processing.
Specific actions to avoid unintended dispersal
The Company should strengthen the operating procedures to ensure that seeds are not
spilled during transport and processing. These requirements should be included in the
consent of the product.

93 Page 25. This and all other page references to EC documents in the answer refer to the page of the
pdf file.
94 Page 1.

576. The full context of the concerns of the UK are given in EC-70/At.74, 25 March 2003, Request
for additional data from UK. The UK is concerne d about accidental release and suggests that the
monitoring plan test the assumption that there will be no spillage, either during importation, transport
or processing.
2. Procedures to avoid seed spill . A central argument used in the environmental
risk assessment of this GMO is the assumption that the rape seeds will not be released
into the environment. However, if there is spillage of seed between import and
processing this may lead to the presence of transient populations of feral GT73
oilseed rape. The UK requests further details fr om the applicant concerning the
standard operating procedures that will be used to ensure that seed is not spilled
during transit. Acceptable procedures to minimize seed spill should be a condition
of any consent issued – this could restrict import and processing to specific locations
and require monitoring for seed spill between the sites of import and processing (see
below).
95
5. Post-market monitoring plan . The proposal for post-market monitoring is not
adequate. As outlined above, the envir onmental risk assessment assumes that seed
will not be spilled, so the post-market monitoring plan should test this assumption.
The UK requests that the applicant giv es further consideration to the post-
market monitoring plan, which should in clude (i) monitoring for the presence of
feral GT73 oilseed rape at the points of import and processing, and between
those locations; and, (ii) proposals for action should feral GT73 be identified.
The proposal for post-market monitoring also includes a request to end users of GT73
grain to inform the regulatory authorities of any adverse effects that are attributable to
the use of the GMO. The UK considers that the applicant should be more proactive
in obtaining this information and should be combined with a reporting requirement.
The UK requests that the consent holder approach end users directly to ask if
they have observed any effects to cov er general surveillance and the results of
this should be reported to the competent authorities at intervals of 6 months for
the first 3 years of the consent. The report should list the requests that have been
made to end users, the proportion that elic it a response and what, if any, unexpected
effects were observed (adverse or otherwise).96
577. These concerns of the UK were initially articulated by the scientific Advisory Committee on
Releases to the Environment (ACRE) in EC-70/At.69, 10 March 2003, ACRE Advice to the Secretary
of State for Environment, Food and Rural Affairs, Scottish Ministers, Ministers of the Welsh
Assembly Government and the Department of En vironment (Northern Ireland). Their advice and
reasoning is repeated here.
Primary Advice:
• 3. The post market monitoring plan should include monitoring for spillage of GT73
seed during import, transportation and processing and test for the establishment of
feral populations of GT73 oilseed rape. The plan should also include appropriate
emergency plans should such populations be identified.

95 Page 2.
96 Page 2.

Post market monitoring plan:
• The post market monitoring plan is a new requirement under Directive 2001/18/EC.
The plan provided by the notifier is brief and limited. On the basis of the risk
assessment for GT73 Monsanto do not propose to carry out any case-specific
monitoring. ACRE agree with this conclusion as the risk assessment does not
identify any risks in relation to human h ealth or the environment from the use of
GT73 oilseed rape as defined in the notification (i.e. for import and processing).
• Directive 2001/18/EC places a requirement to provide a monitoring plan for the
occurrence of adverse effects not anticipated in the environmental risk assessment.
ACRE consider that the proposal in respon se to this requirement put forward by the
notifier is unsatisfactory. In particular the plan does not take into account monitoring
of the occurrence and effects of seed spill during transportation and processing of the
oilseed rape seed. Based on current experience regarding movement of non-GM
oilseed rape in the UK seed spill is likely to occur and will result in the survival and
establishment of feral oilseed rape populati ons and their hybridisation with crop and
other feral populations. It is accepted that the risk of harm to human health and the
environment from seed spill is low, however there is an issue, in the UK, regarding
the segregation of transgenic and non-transgenic material. Hence, ACRE request that
Monsanto review their monitoring plan taking into account methods of monitoring
and reporting seed spill, incorporating a proposal as to how seed spill may be
controlled. It should also include plans for monitoring and controlling establishment
of feral populations as a result of seed spill.
578. In addition, Sweden raised a monitoring issu es in EC-70/At.67, 28 February 2003, Swedish
Board of Agriculture, which is repeated below. Sweden has focused on procedures to report adverse
effects.
A. Monitoring plan
1. The monitoring focuses on the identifi cation, checking and traceability of GT73,
whereby Monsanto Europe is to:
• pass information to the traders and users w ithin the processing industries in order for
them to be able to identify any negative effects on the environment or on human
health and request these users to info rm the Competent Authorities about any
identified negative effects on the environment or human health which are assumed to
have come about from the use of GT73,
• to inform the European food industries dir ectly when the product has been approved
and to request operators in food chains to inform the Competent Authorities about
any identified negative effects on the health of animals, which are reported to them by
farmers or other users, and which are assumed to have come about from the use of
GT73,
• immediately inform the Commission and Competent Authorities during the consent
period if the company receives information about negative effects of the product.

579. Moreover, although Norway is not a part of the EC, Norway submitted comments related to
monitoring in EC70/At.78, 25 March 2003, Comments from Norway. Norway merely notes that there
should be a monitoring plan.
Although required according to Directive 200 1/18/EC, there is no monitoring plan
attached to the application. The Competent Authority of the Netherlands
recommends an approval on the condition that the notifier gives a yearly report of the
sold quantities of line GT73 in each count ry. We will comment on the need for
additional monitoring when we have finished our assessment of the risks for human
health and the environment.
580. Notwithstanding that a monitoring plan is required by Directive 2001/18/EC, it is still
possible to consider if a monitoring plan is necessary from a scientific standpoint, and if so, what kind
of monitoring plan could appropriately address the potential risks.
581. Before doing so, I will establish that in th e SNIF to COM on 16 January 2003, the notifier
proposed a plan that must be considered a case-sp ecific monitoring plan. This is a critical step,
because had the notifier not proposed such a monito ring plan, then the issue would hinge on whether
any monitoring plan is needed. Instead, the i ssue will hinge on the adequacy of the proposed
monitoring plan. This is a substantially more complex argument.
582. In EC-70/At.54 (16 January 20 03 SNIF), the notifier states that unintended release can occur
during import, transport and processing. The notifier suggests that because most grain is crushed near
seaports and modern equipment is used that there is little chance of unintended release.
7. Measures suggested by the notifier to take in case of unintended release or
misuse as well as measures for disposal and treatment
This application is for consent for import and use in the EU of GT73 as any other
OSR, but do not include the cultivation of varieties in the E.U. Therefore an
unintended release would be more likely to occur during import, processing and
transportation of GT73 grain. However, most OSR grain imported into the E.U. via
European sea ports is immediately crushed in nearby crushing facilities and modern
methods of transportation and grain handling minimize losses of grain. Therefore,
there is little chance of germination of grai n resulting in the development of mature
plants of GT73 in the E.U. environment.
GT73 is intended for use by oilseed producers, in the same manner as other oilseed
rape varieties. The measures for waste disposal and treatment for GT73 products are
the same as those for other oilseed rape products.
97
583. In EC-70/At.54 (16 January 2003 SNIF), the notifier argues that there is no need for post-
release monitoring. First, two years of monitori ng volunteers in Canada and the US indicated that
there were no greater numbers of volunteers than non-transgenic rape and that the transgenic
volunteers could be controlled by other herbicides. Second no adverse effects of GT73 have been
identified.
584. Previous commercializations of GT73 have been alongside a stewardship program, which
provides a communication channel if an unanticipated adverse effect would occur.

97 Page 5.

(f) Aim of post-releases monitoring
In the case of the field testing performed in Canada and the US, the fields were
monitored for volunteers during a period of two years and volunteers were chemically
removed.
Since no adverse effects of GT73 have been identified (see section 29), no
requirement for case-specific post-release monitoring is indicated, which is consistent
with approvals granted in other world areas.
GT73 has been commercialized alongs ide stewardship programmes involving
downstream stakeholders in the use of this oilseed rape, in order to ensure the
implementation of good agricultural practice in its cultivation and a channel of
communication in the unlikely event that unanticipated adverse effects might occur.
(g) Duration of post-releases monitoring
In the case of the field testing performed in Canada and the US, the fields were
monitored for volunteers during a period of two years.
(h) Conclusions of post-release monitoring
The monitoring conducted after the Canadian and US field releases showed no
differences in volunteer numbers between GT73 and conventional OSR varieties in
the number of volunteers, and confirmed that GT73 volunteers can be effectively
controlled. No unanticipated effects have been observed since the commercialization
of GT73 in other world areas, nor during th e many years of field-testing inside and
outside the E.U.
98
585. In EC-70/At.54 (16 January 2003 SNIF), the notif ier indicates that the monitoring plan will
focus on general surveillance for unanticipated adverse effects.
D. INFORMATION RELATING TO THE MONITORING PLAN-
IDENTIFIED TRAITS, CHARACTERISTICS AND UNCERTAINTIES RELATED
TO THE GMO OR ITS INTERACTION WITH THE ENVIRONMENT THAT
SHOULD BE ADDRESSED IN THE POST COMMERCIALISATION
MONITORING PLAN
2. Identification of the occurrence of adverse effects of the GMO or its use on
human health or the environment which were not anticipated in the E.R.A.
The environmental and human health safety assessment for GT73 did not identify any
specific risks related to its placing on the market during storage, processing and other
uses. Therefore the monitoring plan for GT73 is focused on general surveillance for
unanticipated, adverse effects.
99

98 Page 19.
99 Page 20.

586. In EC-70/At.55 (16 January 20 03 SNIF), the notifier indicates that there will be no special
measures taken to prevent escape or misuse of the product nor will th ere be any specific instructions
or recommendation for storage or handling.
Part II. Additional Relevant Information
5. The measures to be taken in the ev ent of the escape of the organisms in the
product or misuse of the product.
GTOSR is targeted for use by oilseed producers, in the same manner as other OSR
varieties. Dissemination of OSR plants in to non-agricultural environments, such as
roadsides and other semi-managed areas, occurs currently. The information
presented in this application establishes that the frequency of dissemination of
GTOSR will be similar to other OSR, a nd no special measures will be taken to
prevent dissemination.
6. Specific instructions or recommendations for storage and handling of the
product.
Since GTOSR varieties have b een demonstrated to be s ubstantially equivalent to
other OSR varieties, apart from their tolera nce to glyphosate, no specific instructions
or recommendations for storage and handling are envisaged.100
587. Lastly, the notifier submitted with the 13 December 2002 notification to the Netherlands CA
an Annex VII, Monitoring Plan (EC-70/At.51) in wh ich they propose that no case-specific monitoring
is needed.
588. However, in the same appendix they pro pose three measures to identify potential
unanticipated adverse effects. All three measures are specific to GT73, so these measures must be
considered a proposal for case-specific monitoring, even if the notifier and ACRE (paragraph 577) do
not recognize them as such. The specifics of this proposal follow. It should be clear that although
this proposal parallels the concerns of Sweden (paragraph 578), it is substantially different.
2. Identification of the occurrence of adve rse effects of the GMO or its use on human
health or the environment which were not anticipated in the e.r.a.
Since the notification of GT73 oilseed rape is for consent for import only, and since the
majority of use of this oilseed rape will be for human and animal consumption (see Annex IV,
A.4.), it follows that unanticipated affects are most likely to be manifested as a result of this
use. Therefore, it is proposed:
• To provide traders and processors of bulk mixtures of oilseed rape grain, likely to
contain GT73 oilseed rape grain, with pr oduct information about GT73 oilseed rape
grain. Traders and processors will be request ed to inform the relevant authorities of
any adverse effects on the environment or human health, which they consider to be
attributable to GT73 oilseed rape grain.
• To inform the European feed industry directly, by way of a public announcement, of
the consent for placing on the market of GT73 oilseed rape at such time as it appears

100 Page 66.

in the Official Journal of the rapporteur Member State. Monsanto will also offer to
meet with interested operators to discuss the safety and general characteristics of the
product. Operators in the feed chain will be requested to inform the relevant
authorities of any adverse effects on animal health reported to them through farmers
or national feed associations, which they consid er to be attributable to the feed use of
GT73 oilseed rape grain.
• For the duration of the authorization of GT73 oilseed rape, to immediately inform the
Commission and the Competent Authorities for Directive 2001/18/EC, of any reports
of adverse effects, which come to the atte ntion of Monsanto, so that any reports can
be further investigated by the appropriate authorities.
589. Although it may be true that the notifier made a similar monitoring proposal in the 16 January
2003 SNIF, I could not find this clearly stated in the SNIF. However, under the assumption that any
item in the notification to a country CA is automatic ally a part of the submission to the COM, it must
be concluded that there was a case-specific monitoring plan proposed in the 16 January 2003
SNIF.
590. Are the additional requests made by the memb er nations regarding the monitoring plan
necessary to ensure that conclusions of the safety assessment were valid? To argue this question, I
will first consider if concerns over inadvertent esc ape are valid, then consider if there are valid
reasons for distinguishing GT73 from other rape vari eties, then discuss the ad equacy of the proposed
case-specific monitoring plan, and finally I will addr ess the specific monitoring advice provided by
the various member countries. In addition, I will consider if there was any change in the proposed
monitoring plan, based on an examination of th e new information submitted by the notifier on
5 August 2003 (EC-70/At.92).
591. Concerns that rape seed can escape during import, transport or processing are valid. This was
first brought to scientific attention for rape seed in 1995 in the UK. 101 Moreover, using standard
modern equipment, it is simply not possible to ensu re that spillage will not occur. The concerns of
Denmark (paragraph 571), Austria (paragraph 572), Spain (paragraph 575) and the UK
(paragraph 576) begin with this concern.
592. Should transgenic GT73 be distinguished from conve ntional rape varieties? Here the various
countries express different perspectives. Denmark (paragraph 571), Spain (paragraph 575), the UK
(paragraph 576) and Norway (paragraph 579) do not link their request for additional monitoring to a
specific risk. Austria is concerned about potential contamination of other agricultural systems with
undesired transgenes (paragraph 572), Italy is concerned about potential effects of glyphosate
metabolites (paragraph 574), and Sweden is concerned that a monitoring system is set up so that
adverse effects, should they occur, can be identified (paragraph 578). In addition, the UK (paragraphs
576 and 577) suggests that monitoring should test the assumption of the notifier that seed will not be
spilled.
593. Without a connection to a specific risk or th e risk assessment, requests for monitoring GT73
cannot be distinguished from conventional rape varie ties, and the requests and concerns of Denmark,
Spain and Norway cannot be scientifically justifie d on the basis of the written record received.
Austria's specific concern and the reasoning behi nd the UK requests are scientifically justified, and
both would require that GT73 is distinguished from conventional rape varieties. I was unable to

101 Crawley, M.J. & Brown, S.L. (1995). Seed limitation and the dynamics of oilseed rape on the M25
motorway. Proc. R. Soc. Ser. B.-Bio, 259, 49-54.

determine if Italy's concern had been treated in the notification (13 December 2002) or the SNIF (16
January 2003). If it has not been adequately assessed, then Italy's concern is scientifically justified,
and would distinguish GT73 both from conventi onal rape varieties and from glyphosate itself
(because the plant metabolite AMPA would be less of a concern on non-GMHT oilseed rape crops).
Sweden's concern is that the monitoring plan be sufficient to identify unanticipated adverse effects.
594. Adequacy of proposed case-specific monitoring plan (EC-70/At.51, Annex VII, Monitoring
Plan). The test for adequacy is whether the propo sed plan would meet the aims proposed by Sweden,
Austria, the UK and Italy. The first measure (paragraph 587)102 partially meets Sweden's concern, but
does not address the concerns of Austria, th e UK or Italy. The second measure (paragraph 587)103
meets none of these countries concerns. The third measure (paragraph 587)104 partially meets
Sweden's concern, but does not address the concerns of Austria, the UK or Italy. Thus the proposed
monitoring plan does not meet the concerns of the Sweden, Austria, the UK or Italy.
595. Specific monitoring advice from the member countries is incomplete and too inflexible. By
incomplete, I refer to the criteria for monitoring in my response to question 24a. By inflexible I refer
to the recommendations that the notifier monitor rapeseed cultivation areas, gene dispersal, gene
transfer, studies on transgene expression in wild relatives and conventional oil seed rape crops, and
sampling of volunteers. Meeting the concerns of Sweden, Austria, the UK and Italy do not require
such an intensive monitoring plan. A general prin ciple of GMO risk assessment is that risk is
proportional to the scale of release.
105 Low frequency releases at sma ll spatial scales will have lower
risk, and the monitoring system should reflect this.
596. The revised monitoring plan (5 August 2003, EC-70/At.92, Appendix 9) 106 falls well short of
the meeting the concerns expressed by Sweden, Austri a, the UK and Italy, and meets even fewer of
the criteria for an adequate monitoring plan that the original proposal (13 December 2002, EC-
70/At.51).
597. In EC-70/At.85, the notifier makes an effective argument 107 that grain entering the main
gateways and crushed in the port areas is very un likely to establish in the environment. Hence it
could be suggested that the concerns of Austria, the UK and Italy could best be met by concentrating
monitoring activities in places other than these main gateways and crushing facilities, focusing on
those for which escape may be more likely (worst cases). By monitoring a few potential worst cases,

102 To provide traders and processors of bulk mixtures of oilseed rape grain, likely to contain GT73
oilseed rape grain, with product information about GT73 oilseed rape grain. Traders and processors will be
requested to inform the relevant authorities of any adverse effects on the environment or human health, which
they consider to be attributable to GT73 oilseed rape grain.
103 To inform the European feed industry directly, by way of a public announcement, of the consent for
placing on the market of GT73 oilseed rape at such time as it appears in the Official Journal of the rapporteur
Member State. Monsanto will also o ffer to meet with interested operators to discuss the safety and general
characteristics of the product. Operators in the feed chain will be requested to inform the relevant authorities of
any adverse effects on animal health reported to them through farmers or national feed associations, which they
consider to be attributable to the feed use of GT73 oilseed rape grain.
104 For the duration of the authorization of GT73 oilseed rape, to immediately inform the Commission
and the Competent Authorities for Directive 2001/18/EC, of any reports of adverse effects, which come to the
attention of Monsanto, so that any reports can be further investigated by the appropriate authorities.
105 NRC (National Research Council). (1987). Introduction of recombinant DNA-engineered
organisms into the environment: key issues. National Academy Press, Washington, DC.
106 Pages 17-26.
107 Page 26.

it can be argued that all other cases are likely to ha ve even lower risk. Th e suggestions by Sweden
(paragraph 578) have not been addressed yet.
598. Monitoring is a form of risk management, and might be argued to be independent of the risk
assessment process. However, in this specific case, issues associated with monitoring are intricately
tied to the issues associated with risk assessment, in that the risks associat ed with unanticipated
possible hazards can be reduced to acceptable leve ls with the appropriate monitoring plan, as
suggested by Sweden. For GT73, specific concerns include contamination of other agricultural
systems and possible effects of glyphosate metabolites. Moreover, monitoring can be used to confirm
critical assumptions in the risk assessment (Annex VII, Directive 2001/18/EC), which is suggested by
the UK for this case. Hence, from these perspectives, it can be concluded that a monitoring plan is
necessary to ensure that conclusions of the safety assessment were valid. However, the
monitoring plans advised by the countries are on ba lance overly specific and do not allow the notifier
sufficient latitude to suggest alternative approaches that would address the concerns in ways that may
be more cost-effective.
599. The Member States requested information related to the monitoring plan on 28 February 2003
(Sweden) and 24/25 March 2003 (Austria, Italy, Sp ain, United Kingdom. I have assumed that the
monitoring plan was submitted to COM with the SN IF on 16 January 2003. The earliest date of
submission of this plan was 13 December 2002. Usi ng the later date, it took the Member State 43-68
days to respond with comments. Using the earli er date, this would be 77-102 days to respond
(including the winter holidays). These do not seem to be unreasonable lengths of time for a response.
Bayer Liberty Link oilseed rape (T45 & Topas 19/2)
C/GB/99/M5/2 (EC chronology 72)
Question 29: Given the information before the Pa nel, including the notification (EC-72/At.1-3)
and additional information provided by AgrE vro (EC-72/At.7-9), were requests for more
information (EC-72/At.5, 6, and 11-13) concerning feeding studies, chemical compositional data
and potential interactions of transformation ev ents in the stacked product necessary to ensure
that conclusions of the safety assessment were valid?
Dr. Nutti
600. Based on the information provided by the applicant in EC-72/1-3 and additional information
at EC-72/At.7-9 and the minutes of the Sixty-Seco nd Meeting of ACRE, when it was verified that
there were a number of inconsistencies in the mo lecular data provided and the dossier was rather
impenetrable. Also, too much important material was buried in annexes rather than being in the core
dossier and the descriptions of the plasmid maps and analyses did not correspond with the data as
presented. Further concerns were expressed about de ficiencies in the molecular studies carried out to
characterize the inserted DNA, although it was accepte d that the appropriate experimental data may
have been supplied somewhere in the notification dossier but it was not immediately obvious where it
might be. Therefore, I understand that ACRE and the UK SC requests for more information were
necessary to ensure that conclusions of the safety assessment were valid. It is important to point out
that ACRE was not being asked for formal advi ce at that stage, only guidance; therefore, no
conclusions were drawn at the time.

Stoneville BXN cotton (10215, 10222, 10224) (formerly held by Aventis and Calgene)
C/ES/99/01 (EC chronology 73)

Question 30: Given the information before the Panel, including the notification (EC-73/At.1),
was additional information regarding environmenta l impact tests, feed safety, residue analysis,
monitoring for herbicide resistance requested by Spain (EC-73/At.3) necessary to ensure that
conclusions of the safety assessment were valid?
Dr. Nutti
601. This question could not be properly answered because the complete notification was not made
available to me (At. 1 corresponds to the notific ation index only). However, the answers to the
questions, which are presented in At.4, seem to be satisfactory as far as food safety is concerned
(question 7). Nevertheless, the entire evaluation pro cess is, by the chronology made available, at its
initial stage, so conclusions on the validity of the safety assessment cannot be drawn.
Dr. Andow
Necessity to ensure validity of safety assessment
602. I will address items 8, 9 and 11 as listed in EC-73/At.03 (15 July 1999, proposed amendments
to 3 May 1999 notification made by lead CA). These relate to environmental impact tests, herbicide
studies, and resistance evolution. The issue is to determine if this additional information is necessary
to ensure valid conclusions of the safety assessment.
8. The results concerning the environmental impact of these genetically
modified plants refer almost exclusively to studies conducted in the USA, Japan,
Argentina, etc. (page 37). The results of tests conducted in Spain and Greece should
also be included, considering that in some cases the environmental conditions cannot
be extrapolated.
9. Indicate where and when the survival study described on page 38 was carried
out. These studies were conducted on two lines other than the lines covered by this
application.
11. Provide toxicological and ecotoxicological studies on the herbicide used on
this type of crop.
13. As regards points 11 and 12, the results of the studies must be indicated as a
function of the different doses of product applied, as was the case in various
phonological studies on cotton plants.
In addition to clarification of these points, the National Biosafety Committee
considered that the company should pr opose specific methods for management,
monitoring and handling of this crop in order to reduce the possibility that resistance
to this herbicide could emerge.
603. Similar concerns are reiterated in EC-73/At. 12, the 26 September 2003, analysis of the 16
January 2003 notification by the Spanish NCB.

these conclusions do not address how much additional information would be necessary. Given the
information before the Panel this quantitative issue cannot be addressed.
612. The information requested by Spain was on 15 Ju ly 1999 to material submitted by the notifier
on 3 May 1999. This was 73 days to respond. This does not seem unreasonably long.
Question 31: Given the information before the Panel, including the notification and the
notifier's responses to questions (EC-73/At.2), was additional information regarding feed safety
and compositional analysis (particularly related to nitrilase levels) (EC-73/At.3) necessary to
ensure that conclusions of the safety assessment were valid?
Dr. Nutti
613. Once again, this question could not be prope rly answered either, since the complete
notification was not made available to me (At.1 co rresponds to the notification index only). However,
the answer to question 7 (nitrilase le vel in cottonseed oil) seems to be satisfactory. Also in this case,
the entire evaluation process is, by the chronology made available, at its initial stage, so conclusions
on the validity of the safety assessment cannot be drawn.
Pioneer/Dow AgroSciences Bt corn Cry1F (1507)
C/NL/00/10 (EC chronology 74)

Question 32: Given the information before the Pa nel, including the original notification (EC-
74/At.1-15), was the information regarding molecular characterization of this product requested
by the lead CA (EC-74/At.17 and 33) necessary to ensure that conclusions of the safety
assessment were valid?
Dr. Nutti
614. Based on the information provided by the applicant (EC-074/At. 01-15) and the request by the
lead CA (EC-074/At.17), I can conclude that a lot of information was provided.
However, I do not feel capable of judging if such material was enough to ensure that conclusions of
the safety assessment were valid.
Question 33: Given that Pioneer had notified this product under the regulation concerning the
deliberate release of this plant (rather than the novel food regulation), were concerns raised by
the lead CA about allergenicity of the truncated synthetic Cry1F protein relevant to the CA's
assessment of this product (EC-74/At.17 and 33)?
Dr. Nutti
615. Based on the information provided by the applicant (EC-074/At. 01-15) and the specific
request on allergenicity by the lead CA (EC-074/At.17) and checking EC-074/At.05, page 54, my
understanding is that the applicant provided information on the history of safe use of the donor
organism and a general explanation on the other ite ms, such as molecular weight of the protein,
protein degradation in gastric fluids, expression levels of the protein and glycos ylation of the protein.
I believe that the applicant was supposed to pres ent experimental data to support this general
information.
616. Based on the further information presented by the applicant (EC-074/At.19) I can assume that
the work carried out by Evans (1998) provides fundamental information on protein equivalence

(maize expressed core CRY1F and microbially-derived trypsinolyzed core CRY1F). However, I could
not recover such information, presented on tables 10A, 11A, 12A and 13A in EC-074/At. 19) (I could
not open the files on my computer). So, I cannot assess if the request from the lead CA was fulfilled.
617. From EC-074/At.33, I can infer that the se cond package of information provided by the
applicant seems not to be sufficient as far as experimental data are concerned.
618. Examining EC-074/At.36, once again no comple te annexes could be found (only pages
identifying each annex can be visualized, but not the complete papers which were supposed to be
there).
619. From all the above exposed, I do not have enoug h information to conclude that the concerns
raised by the lead CA about allergenicity of the tr uncated synthetic Cry1F protein are relevant to the
product's assessment. I would like to point out that th is specific process called my attention since it is
completely different from others I have reviewed, i. e., in my opinion, the requested information is not
provided in a straight way, and experimental data are not presented in a clear way.
620. Based on EC-074/At.060, the comments from RIKILT on the work by T. Meyer (1999)
concerning the need of correct updated data, full description of the methodology, and significant
homology (according to FAO/WHO Expert Consulta tion on Allergenicity of Foods Derived form
Biotelchnology, 2001) are, in my opinion, correct.
Question 34: Given the information before the Panel, including the original notification and
additional information provided by Pioneer (EC-74/At. 18-32, 35-50, and 53-59), was the
information regarding feed safety, detection me thods and reference materials requested by the
lead CA (EC-74/At.52 and 60) necessary to ensu re that conclusions of the safety assessment
were valid?
Dr. Nutti
621. Based on EC-074/At.052_SCI, Annex 1B, the request from the lead CA [xxx] was necessary.
622. Based on EC-074/At.052_SCI, Annex 2B, the comments from RIKILT [xxx] are correct.
623. Based on EC-074/At.052_SCI, Annex 6B, the comments from RIKILT [xxx] are correct.
624. Although I could not find the quail feeding study within the files, I would like to point out
that, according to the Codex Guideline for the Condu ct of Food Safety Assessment of Foods Derived
from Recombinant-DNA Plants (Page 4, paragraph 53) , some foods may require additional testing;
regarding animal feeding studies, extra studies may be warranted if changes in the bioavailability of
nutrients are expected or if the composition is not comparable to conventional foods.
625. Examining EC074/At.058_SCI, where the detection method is presented, so far as my
knowledge goes, it seems to be correct (in my opi nion, the notifier has to submit the method, the
primers sequences and the reference materials, wh ich was done). Although in EC074/At.060 the lead
CA and RIKILT keep the same requests and comments , I do not feel capable of judging the raised
issue of intellectual property and confidentiality.
Question 35: In your view can EC field trials, in France, Italy, and Chile, provide compositional
data on maize kernels that is relevant to evaluating cultivation areas exporting maize to the EC?

Dr. Nutti
626. It is not clear to me whether the mentioned EC field trials have already been done or are being
requested. If they have already been carried out, the Codex Alimentarius Guidelines for the Conduct
of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003,
Para 45) states that the location of the trials sites should be representative of the range of
environmental conditions under which the plant varieties would be expected to be grown. The number
of trial sites should be sufficient to allow accurate assessment of compositional characteristics over
this range. Similarly, trials s hould be conducted over a sufficient number of generations to allow
adequate exposure to the variety of conditions met in nature. Therefore, in this case, I understand that
the field tests in EC and Chile can be considered as supplementary information to the previous tests
carried out by the notifiers, although they were not necessary, (specially when we remember that the
maize in question if for importation and processing).
Dr. Andow
627. Clarification from the Panel: Information provid ed by the notifier makes reference to field
trials conducted in Chile that (it argues) provide compositional data on maize kernels that would be
relevant to the evaluation of [?] products exported to the EC. See the notifier's response to questions
from the CA, EC74/At.52_SCI, page 6.
Relevance to EC maize imports:
628. The question with the response from the notifier is in EC-74/At.52_SCI, 7 December 2001
(question originally posed 19 March 2001, EC-74/At.33), as follows.
Question 12
The applicant is requested to supply com positional data for kernels harvested from
geographical areas representative for the co mmercial cultivation of maize 1507 (prior
to export to the EU). Data for such areas should include multiple locations within an
area and cover at least two seasons. In addition, these data should include the
following treatments: transgenic 1507 ma ize plants treated with glyfosinate,
untreated transgenic 1507 maize plants, and untreated non-transgenic control plants.
Pioneer's reply to question 12
The locations for the field experiments th at are described in the dossier, namely
France, Italy, and Chile, would be repr esentative for the cultivation areas exporting
maize to the EU. GM maize plants cultivat ed at the EU test sites had been either
treated or not treated with the herbicide glufosinate.
Comment RIKILT
RIKILT has not been convinced by Pioneer's statement that the EU locations would
be representative of non-EU locations exporting maize and maize products to the EU.
629. It is clear that France and Italy cannot be locations representative of areas exporting maize to
the EU, as they are part of the EU.

630. World maize export and import statistics are available from the US Department of Agriculture
Economic Research Service. These statistics are summarized below.
World Corn Trade
Thousand Metric Tons
Date Created: 12/13/2004
 2000/01 2001/02 2002/03 2003/04 2004/05
Exports December
Argentina 12,229 8,581 12,349 10,400 10,000
Brazil 3,741 3,857 3,181 5,818 3,000
Bulgaria 15 41 191 100 300
China, Peoples Republic of 7,276 8,611 15,244 7,553 4,000
EU-25 1,016 2,849 1,995 400 500
Paraguay 386 262 516 800 600
Romania 50 135 144 100 1,000
South Africa, Republic of 1,415 1,182 1,141 797 1,000
Serbia and Montenegro 50 21 200 50 600
Thailand 407 184 137 726 450
Ukraine 397 349 811 1,238 2,000
Others 1,147 1,201 1,238 1,822 1,165
Subtotal 28,129 27,273 37,147 29,804 24,615
United States 48,329 47,271 40,924 48,645 51,000
World Total 76,458 74,544 78,071 78,449 75,615
Imports
Algeria 1,500 1,537 1,643 1,750 1,800
Belarus 169 126 43 150 150
Brazil 671 297 521 677 400
Canada 2,843 4,022 3,846 2,039 2,200
China, Peoples Republic of 89 39 29 2 200
Chile 1,362 1,278 933 1,000 1,000
Colombia 1,857 1,911 2,112 2,100 2,200
Costa Rica 513 463 514 585 600
Cuba 119 292 279 475 450
Dominican Republic 968 1,038 906 824 1,000
EU-25 3,800 3,801 4,327 5,600 2,500
Ecuador 149 309 304 475 300
Egypt 5,268 4,905 4,848 3,800 4,300
El Salvador 469 287 394 475 450
Guatemala 549 584 513 500 550
Honduras 252 217 214 230 200
Indonesia 1,280 1,149 1,633 1,350 1,300
Iran 1,265 1,261 2,157 1,700 1,900
Israel 993 1,021 776 1,400 1,000
Japan 16,340 16,395 16,863 16,781 16,800
Jamaica & Dep 221 241 218 225 225
Jordan 454 439 406 400 450
Kenya 700 20 82 100 400
Korea, Democratic Peoples Rep 688 288 144 80 200
Korea, Republic of 8,743 8,621 8,786 8,783 8,500
Lebanon 184 263 131 285 250
Libya 252 235 134 150 200
Malawi 50 86 25 150 200
Morocco 966 829 1,054 1,200 1,200
Mexico 5,928 4,076 5,269 5,707 5,800
Malaysia 2,588 2,425 2,408 2,100 2,000

 2000/01 2001/02 2002/03 2003/04 2004/05
Mozambique 40 369 413 100 200
Peru 861 858 917 1,100 1,000
Panama 249 237 272 300 300
Zimbabwe 50 311 625 600 600
Russian Federation 150 534 99 500 600
Saudi Arabia 1,389 1,268 1,424 1,550 1,600
South Africa, Republic of 0 726 617 495 200
Syria 794 892 919 950 1,100
Tunisia 776 793 734 800 800
Turkey 608 1,193 1,475 1,050 900
Taiwan 4,924 4,661 4,681 4,900 4,700
Venezuela 1,207 515 675 680 800
Yemen 200 264 252 200 200
Others 2,872 2,203 2,166 2,022 1,790
Subtotal 75,350 73,279 76,781 76,340 73,515
Unaccounted 929 1,063 916 1,768 1,720
United States 179 202 374 341 380
World Total 76,458 74,544 78,071 78,449 75,615

Source: USDA Official Estimates http://www.fas.usda.gov/psd/complete_tables/GF-table7-88.htm

Note:
 – The international trade year for corn is October/September.
 – 'Unaccounted' includes grain in transit, reporti ng discrepancies in some countries, and trade to
countries outside the USDA database.
 – This table was prepared by the Grain and Feed Division, Commodity and Marketing
Programs, Foreign Agricultural Service, USDA, Washington DC 20250. Information is
gathered from official statistics of foreign governments and other foreign source materials,
reports of U.S. agricultural att aches and Foreign Service officer s, results of office research,
and related information.

631. It is clear that the US, Argentina, People's Republic of China and Brazil are major exporters
of maize.
632. From a statistical perspective, some reasonable representation of actual imports to the EU
would seem necessary. There has been no attempt to do this. Indeed there seems to be a dismissive
attitude to the request.
633. It can be asked if maize from Chile would in fact be representative of maize from other
regions of the world. It is likely that for some industrial uses of maize, that there is little need to
distinguish among regional sources. Without data supporting this lack of need to distinguish
geographic sources, it is not possible to assert that Chile would be representative of all importation
regions.
634. Hence, maize from France, Italy, and Chile wo uld not provide compositional data on
maize kernels that is relevant to evaluating cultivation areas exporting maize to the EC.
635. Even though the question was not asked by the Panel, the scientific rationale for the request
for compositional data is not evident from the written record. It would seem essential to provide such
a rationale concomitant with the request for inform ation from the notifier. More specifically, while I
could imagine some reasons for such a request, many of these imagined reasons are NOT necessary
for completing a scientific risk assessment.

Pioneer/Dow AgroSciences Bt corn Cry1F (1507)
C/ES/01/01 (EC chronology 75)

Question 36: Given the information before the Panel, including (EC-75/At.1), was the
information regarding molecular characterization , toxicity, and environmental impact of this
product requested by the National Biosafety Committee of Spain (NBC) (EC-75/At.5) necessary
to ensure that conclusions of the safety assessment were valid?
(a) Given that Pioneer had notified this product under the regulation concerning the
deliberate release of this plant (cultivation), were concerns raised by the lead CA
on allergenicity and toxicity of the pr oteins expressed by the inserted gene
sequences relevant to the risk assessment of this product?
Dr. Nutti
636. Based on EC-075/At.1, regarding Question 5 on protein expression, the protein used in the
toxicology and allergenicity tests was obtained in a heterologous system and not in other maize. Thus,
the notifer provided the correct information, which the lead CA did not understand. The possible
reason for the misunderstanding resides in the fact that the same protein is expressed in maize 1507
and in maize 1360. But, it is important to point out that the studies were carried out with protein
derived from recombinant microorganisms.
637. Based on EC-075/At.1, regarding Question 6 on protein toxicity and allergenicity, the lead
CA asks about products which clearly degrade duri ng processing. The answer provided by the notifier
is very well detailed and comprehensive, clarifyi ng all the topics regarding the safety assessment of
protein toxicity and allergenicity. My impression is that the lead CA was not the most suitable person
to evaluate the case.
Dr. Andow
Necessity to ensure validity of safety assessment:
638. I will cover the toxicity and environmental impa ct parts of this question. I will address
whether the information requested by the NBC was necessary to ensure that the conclusions of the
safety assessment were valid.
639. The specific information requested by the NBC that is relevant to the part of the question I
will address is detailed in EC-75/At.05, 28 November 2001, and repeated below.
Field trials in Spain
1. In accordance with its rules and pr inciples, the National Biosafety Committee
considers it necessary to conduct and have the results of experimental trials in Spain
before proceeding to examine the applicati on to place the product on the market or
for the company to give sufficient reasons wh y there is no need to conduct such trials
in Spain.

Protein expression
5. The toxicity and allergenicity tests for CRY1F and PAT proteins must be
carried out on proteins obtained from 1507 maize, not from 1360 maize which,
although obtained in the same way, is a different transformation event.
6. Point 4.3(ii) on page 134 states that irrespective whether or not the process
degrades the inserted sequences and the CRY1F and PAT proteins, the effects of the
product on animal feed are considered in significant since neither the CRY1F and
PAT genes nor the cry1f and par proteins are toxic or allergenic. This claim is
debatable, above all with regard to the possible toxicity of degradation peptides
compared with full proteins. Without going any further, Bt protein itself is toxic
when processed. It would therefore be desirable to check that technological
processing of grain and forage derive d from 1507 maize leads to no specific
degradation of the proteins analysed. If it does, it would be necessary to analyse the
allergenicity or toxicity of the specific degradation products.
Environmental impact
9. The information provided by the company on studies is insufficient,
particularly on the impact on other species and/or the environment. There are many
cross-references to unpublished documents. In order to evaluate the experiments
conducted, the following documents are needed: Hoxter et al. 1999 a, b, c and d;
Higgins, 1999; Maggi, 1999; Halliday, 1998b.
10. Given that the Mediterranean corn borer (Sesamia nonagrioides) is the
greatest pest in Spain, data are needed on th e efficacy of this transgenic maize against
this species in the laboratory and in the field (vernier 2001b).
11. Annex 1, Appendix 1 includes no non-target species of relevance to Spain.
Species which are endangered, sensitive to ch anges in their habitat and of special
interest should be covered. More specifi cally, species occurring at the same time or
place as maize pollination must be taken into consideration.
12. The resistance management plan proposes voluntarily reserving 5% of the
area as a refuge with non-Bt maize, but fails to state that insecticides must not be
applied within this area so that it can ser ve as a true refuge. The monitoring plan
must be brought into line with the legislation in force.
640. After receiving responses from the notifier, the NBC amplified their request for experimental
trials in Spain by requesting additional trials, and evaluations of he rbicide efficacy (EC-75/At.13, 17
June 2002). While these requests are not included in the question at hand, they suggest that request 1
in EC-75/At.05, 28 November 2001, was considered quite important by the NBC.
Field trials in Spain
1. After examining the additional information supplied by Pioneer, the National
Biosafety Committee repeated that more experimental trials were needed in Spain and
that the results should be placed at its dispo sal in order to ascertain the behaviour and
efficacy of this maize under the different environmental conditions which can occur

in Spain and its impact on populations of co rn borer and of other non-target species
associated with maize crops in Spain.
2. Data are also needed on evaluations of the efficacy of the herbicide on this
crop under conditions in Spain, since this c ould depend on the climate in each area or
region.
641. The original notification submitted by the notifier on 6 July 2001 was not included in the
materials. EC-75/At.01 appears to be the original Spanish version of EC-75/At.06-11. This means
that I cannot assess the necessity of the request in th e context of information already provided in the
original notification. Instead, I will have to judge the necessity of the requested information on the
presumption that the answers provided by the notifier on 14 February 2002 109 to questions 5, 6, 9, 10,
11 and 12 of EC-75/At.05 were sufficient to m eet the information needs requested. On this
presumption, I can determine if the information provi ded was necessary to ensure that conclusions of
the safety assessment were valid. As Question 1 apparently was not sufficiently addressed in the 14
February 2002 response, and apparently was still unresolved over a year later (EC-75/At.21, 17
February 2003), I believe too much time and additio nal information intervened to judge the necessity
of the original question on the basis of an analysis of the response. I will evaluate this question
without reference to the answers provided by the notifier.
642. Question 5. Question 5 was necessary to ensure that conclu sions of the safety assessment
were valid. It is possible that microbially derived Cry1F protein or Event 1360 derived Cry1F or
PAT are not the same chemical products. If they were not the same chemical products, then results
from toxicity tests could be different, leading to e rroneous conclusions in the safety assessment. It is
possible that identical gene sequences in the pl asmid vector could give rise to different gene
sequences incorporated in the plant. This has been seen for the cry1 Ab gene that was incorporated
into Mon810 and Bt11.110
643. Question 6. Question 6 was necessary to ensure that conclu sions of the safety assessment
were valid. It is possible that stable or transiently quasi-stable degradation products of Cry1F or PAT
proteins could have effects, either by themselves as degradation products or in reacting with other
chemicals, such as sugars, even when the parent compound does not react with other chemicals. I
believe stable or quasi-stable degradation products of Cry proteins in humans have not been found,
however, such products are known in insects. Indeed stable degradation products are the basis of the
toxicity of some Cry proteins to some insects. Hence this possibility cannot be excluded in other non-
mammalian degradation processes. The possibility of degradation products reacting to form
chemicals with some adverse effect is a hypothetical possibility. However, the chemical reactions
that occur in grain processing are complex and fully understood. It should also be recognized that the
possibility of such effects may be small.
644. Question 9. Question 9 was necessary to ensure that conclu sions of the safety assessment
were valid. Actuality a risk assessor can choose to belie ve the interpretation of the data provided by
the notifier or to examine the data and come to an independent interpretation. The scientifically sound
approach would be to examine the data and come to an independent interpretation. The question is
both reasonable and essential.
645. Question 10. This question is difficult to unders tand. Vernier (2001b) is referenced in the
question. This report, which was included in the orig inal notification, evaluates lab and field efficacy

109 EC-75/At.01 (English).
110 Essential Biosafety. www.essentialbiosafety.com/main.php.

of Event 1507 to Sesamia nonagrioides. The questio n asks for lab and field evaluations of efficacy to
S. nonagrioides. Hence, from a literal interpretation of the question, it was not necessary to ensure
that conclusions of the safety assessment were valid. Even if we assume that the question was
requesting field data from Spain, the notifier's answer clearly states that the field data were collected
in an area of southern France, and the NCB did not question this in their follow-up questions. Hence,
is must be concluded that they decided also that the question was not essential, as acceptable data had
already been submitted in the original notifica tion. The only way that this question could be
considered essential is if the data provided by th e report by Casteñera (2001), cited in the notifier's
response filled some significant gap in the Vernier (2001b) study. As neither of these reports were
included in the information before the Panel, it cannot be concluded that the Casteñera (2001) report
filled a crucial gap in the Vernier (2001b) study, and it cannot be concluded that the question was
essential.
646. Question 11. Question 11 was necessary to ensure that conclusions of the safety
assessment were valid. The original notification suggested th at most susceptible Lepidoptera would
not be adversely affected by Event 1507 because exposure levels would be low and the expected
hazard was also low. This part of the risk assessment then presume d that all Lepidoptera of potential
concern would not be adversely affected. The qu estion was necessary to ensure that the specific
Lepidoptera (butterflies) of special concern would not be adversely affected.
647. Question 12. There is legitimate scientific debates that bear on the necessity of this question.
One perspective is that a 5% refuge is insufficient for corn borers regardless of how it is treated and
that a 20% refuge is essential, but this 20% refuge can be treated with other insecticides. From this
perspective, it is the area of refuge that is mo st important as long as the refuge produces some number
of susceptible moths,111 and Question 12 is not justified because its premises are not valid. The other
perspective is that a 5% refuge may be suffici ent if enough corn borers are produced from it. 112 In
other words, refuges should be managed to produce as many susceptible moths as possible. From this
perspective, question 12 was necessary to ensure that conclusions of the safety assessment were
valid. In either case, discussion of various management options for the refuge is necessary.
648. Question 1. The degree that a field trial shoul d be conducted in the actual location of concern
is of considerable scientific interest and there is not yet a scientific consensus on this issue. Whether
data from France and Italy can be used to inform a risk assessment in Spain in a scientifically
definitive manner is difficult to answer in the abstract. In general, it has been appreciated for some
time that climate constrains ecological communiti es and ecosystems creating similarities among them
even when they are in widely disparate regions of the world. More recently, climate has been shown
to similarly structure some biological communities, especially in comparisons of communities in the
world's deserts. Thus, there are reasons to believe that data from France and Italy can be used to
inform a risk assessment in Spain. At the same time, it has been widely appreciated that local
ecological communities and ecosystems often func tions quite differently even from nearby
superficially similar communities and ecosystems. For example, for maize en tomology in the US, it
is unlikely that an extension entomologist in Illinois would accept important results from Iowa,
Indiana or Ohio as accurate for Illinois without doing some experiments to confirm their applicability.

111 This perspective has been most recently advanced by Ives and Andow, 2002, Evolution of resistance
to Bt crops: Directional selection in structured environments. Ecology Letters 5:792-801. The basic idea is that
selection is related mostly to the proportion of refuge moths that are exposed to selection on the Bt crops, which
is proportional to the relative area of the Bt crops.
112 This perspective has been widely suggested, but still has not been proved. The underlying metaphor
is that the refuge moths dilute out resistance in the Bt crops, and the more refuge moths, the greater the dilution
effect. More moths can be produced by producing more moths per unit of refuge or by having larger refuges.

Thus, there are reasons to believe that data from France and Italy may not be used to inform risk
assessment in Spain, unless some of the critical inform ation is confirmed to be relevant for Spain in
experiments in Spain. Based on the exact wording of the question, it cannot be determined which of
these two perspectives would hold in this case. In pa rt, the reason for this indeterminacy is that the
question is so general that it coul d be referring to almost any aspect of field experimentation. The
question should be worded more sp ecifically, both to guide the registrant in answering the question,
and to be clarify the specific purpose of the question. As a consequence, while I would conclude that
questions along the lines of question 1 would be ne cessary to ensure that conclusions of the safety
assessment were valid, in this case, I must conclude that question 1 was not necessary to ensure that
conclusions of the safety assessment were valid.
649. Summary. With respect to toxicity and environmental impacts, questions 1 and 10 were not
necessary to ensure that conclusions of the safety assessment were valid, while questions 5, 6, 9, 11
and 12 were necessary to ensure that conclusions of the safety assessment were valid.
650. Following from this summary, given that th e notifier had notified this product under the
regulation concerning the deliberate release of this plant (cultivation), the concerns raised by the lead
CA on toxicity of the proteins expressed by the in serted gene sequences were relevant to the risk
assessment of this product.
651. The notifier submitted the original notification to the Spanish CA (NBC) on 6 July 2001. The
NBC requested additional information on 30 Oc tober 2001 and 28 November 2001. The notifier
responded to these requests on 14 February 2002. Th e NBC made additional requests for information
on 17 June 2002. From the time of submission to the first request for information 116 days passed.
From the time of submission of responses to the re quests for information to the 17 June 2002 request
for additional information, 92 days passed. This was 208 days total time, and seems longer than
necessary to complete the reviews and make requests for information.
Monsanto Roundup Ready corn (NK603)
C/ES/00/01 (EC chronology 76)

Question 37: Given the information before the Pa nel, including the notification (EC-76/At.1-2
and 27), was further information regarding mol ecular characterisation, nutritional analysis,
and environmental impact requested by the le ad CA (EC-76/At.6) necessary to ensure that
conclusions of the safety assessment were valid?
Dr. Nutti
652. Based on EC-76/At.1-2 and 27, and regarding nutritional analysis, com positional analysis of
NK603 maize grain has demonstrated substantial equivalence with traditional maize. This is
supported by lack of differences shown in the results of feeding NK603 to broiler chickens compared
with birds fed with NK603 parental line or with fi ve commercial reference lines, as well as repeated
dose feeding studies in rats, which contribute to determine the composition and nutritional
equivalence. Whatever studies were further requested by the lead CA in EC-76/At.6, such studies
were not necessary to ensure that conclusions of the safety assessment were valid since all the
relevant information had already been provided. In EC-76/At.27, the Spanish Biosafety Commission
had already estimated that for the uses considered and at the current scientific and technical "state of
the art", there was no scientific evidences which indi cated any risk for human and animal health of
NK603 maize.

Dr. Andow
Necessity to ensure validity of safety assessment:
653. I will cover the environmental impact aspects of questions 37 and 38.
654. The relevant questions from the lead CA are in EC-76/At.5, 15 February 2001 and are
reproduced below.
Environmental impact of the GMO
Although this is not the direct purpose of th is notification, the application covers
aspects relating to the imminent request for authorization for cultivation in another
Member State. In view of this eventua lity, the National Biosafety Committee raises
the following questions:
16. Page 14, Section 3 – Survivability: The comments on plant survival are
incorrect, given that crop repetition is common in many areas and that, in many cases,
grain from fallen ears germinates to produce plants in the next season.
17. Page 16, Section 7 – Potential interactio ns: This point must be looked into as
broadly and deeply as necessary. The environmental impact of these interactions on
target or non-target flora and fauna must be evaluated more thoroughly.
18. Page 58, Point D.6. – Transferability of genetic material from the genetically
modified plant to other organisms: Alt hough the application refers exclusively to
authorization of the grain, detailed knowledge of the dispersal capacity of the pollen
under various conditions is n ecessary and would be of relevance for authorization of
cultivation in the State concerned by the a pplication. This information could be
decisive when the time comes to draw up a future monitoring plan.
19. Page 98 – Appendices: Doses and conditi ons for application of the herbicide
(phenological condition of the plant, dose and date).
655. It is clear from the opening paragraph of these questions that the lead CA justifies the
questions on the basis of an imminent request fo r authorization for cultivation in another Member
State. Thus, I conclude that the lead-CA does not believe that the questions on environmental impact
are necessary to ensure that conclusions of the safety assessment were valid.
656. I believe that none of the questions posed under envi ronmental impact are necessary to
ensure that conclusions of the safety assessment were valid.
657. Given that the notification is not for cultivati on, whether survival is slightly better in
continuous maize than in rotated maize ignores the bigger point, which is that maize does not survive
very well. While the CA is correct in noting the di fference, this difference is not necessary to ensure
that conclusions of the safety assessment were valid.
658. While it is true that potential interactions need to be looked into broadly and deeply if the GM
crop were to be cultivated, it is not true that such an investigation is necessary for the present
notification. It would be bette r to focus attention on detecting accidental releases and quickly
eliminating them.

659. Although it is true that detailed information on transferability of genetic material from the
GM crop to other organisms is needed if the GM crop were to be cultivated, it is not true that detailed
information is needed for the present notification. Some information is necessary to consider how
gene escape can occur either during processing, storag e or transport, but detailed information is not
necessary.
660. As the GM plant is not to be cultivated under the present notification, information on
herbicide application is not needed.
661. From the time that the notification was received (2 January 2001) to the time that the lead CA
sent questions for clarification to the notifier (15 Fe bruary 2001), 44 days had elapsed. This seems to
be a rapid turn around time.
Dr. Squire
662. The weakness of suitable criteria is again at issue here (Notes, paragraphs 2, 7, 8).
Documentation from the company is least on environmental matters, partly because of changed
context (what might be an acceptable impact in one part of the world is considered unacceptable in
another), while the counter-arguments take no st ep towards indicating what standards might be
applied.
Question 38: Given the information before the Panel, including the notification and letter from
Monsanto providing additional information (EC-76/At.7-9), was additional information
necessary regarding molecular composition a nd environmental impacts associated with
accidental germination requested by the lead CA (EC-76/7-9 and 10) necessary to ensure that
conclusions of the safety assessment were valid?
Dr. Andow
663. Having received answers from the notifier (5 September 2001), the lead CA requested
additional information. On the issue of environm ental impacts, the lead CA asked the following
question.
• Although cultivation of this maize is not c overed by this application for marketing
authorisation, details of the potential environmental impact of any accidental
dissemination or germination are needed.
664. Based on the SNIF (EC-76/At.2, 4 August 2000) and the responses to the first set of questions
from the lead CA (EC-76/At.7-8, 5 September 2001), it is clear that the notifier has not addressed this
question. The notifier believes, probably rightly, that the likelihoo d of accidental dissemination and
germination (exposure to the environm ent) is small. If this is true, the notifier is arguing that when
exposure is small, risk is small. Consequently, the notifier may believe that it was not necessary to
address this question.
665. In either event, the lead CA may reason as follows: if the hazard associated with a rare
exposure event is large, then the risk may be large. Hence the question is necessary to ensure that
conclusions of the safety assessment are valid. However, it would be helpful to the notifier to
specify that the lead CA is concerned about large potential environmental impacts. Moreover, it
would be even more helpful to the notifier to suggest some possibilities. For example, if
contamination of conventional produc tion (related to the "coexistence" issue) is a major concern (and

is considered an environmental impact), the notif ier would be able to propose how the concern could
be managed, thereby facilitating the more rapid completion of the notification process.
666. The time from the responses to the first set of questions from the lead CA (EC-76/At.7-8, 5
September 2001) to the time of the second set of questions from the lead CA (EC-76/At.10, 10
October 2001) was 35 days. This seems to be a rapid turn around time.
Dr. Squire
667. On the more specific question of whether GMHT maize persists (accidental germination), it is
justified to query the original statement since emergence has been recorded in some southern
European areas.
Question 39: Given the information before the Panel, including the notification and additional
letter from Monsanto providing additional information (referenced above and EC-76/At.11-12),
was additional information regarding molecular characterization and toxicology requested by
the lead CA (EC-76/At.14) necessary to ensure that conclusions of the safety assessment were
valid?
Dr. Nutti
668. Based on EC-76/At.11-12 and the request by the lead CA in EC-76/At.14, regarding to animal
feeding, asking for clarification of the titles of ta bles 2 and 3 on page 24 of Appendix 2 and of their
content with regard to the calculation of the saf ety margins established and the reasons for the
differences detected, my opinion is that such a reque st was not necessary to ensure that conclusions of
the safety assessment were valid.
Dr. Squire
669. See answer to question 37.
Question 39 bis: Given the information before the Pa nel, including the notification and
additional letter from Monsanto providing additional information (previously referenced and
EC-76/At.11-12), was additional information regarding allergenicity studies and PCR tests
requested by Austria (EC-76/At.44) necessary or u seful to ensure that conclusions of the safety
assessment were valid?
Dr. Nutti
670. Answer 39 bis: Based on EC-76/At.11-12 and the request by the lead CA in EC-76/At.44,
regarding to allergenicity studies and PCR tests, my opinion is that such a request was not
necessary to ensure that conclusions of the safety assessment were valid.
Monsanto Roundup Ready corn (GA 21)
C/ES/98/01 (EC chronology 78)
C/GB/97/M3/2 (EC chronology 85)
Question 40: Given the information before the Panel, including the application
(EC-85/At.25-26), questions by Denmark (EC-85/At.32) and responses to these questions
(EC-85/At.41), was the additional information requested by Denmark (EC-85/At.42) necessary
to ensure that conclusions of the safety assessment were valid?

Dr. Nutti
671. Based on the information provided by the applicant (EC-85/At.25-56) and responses to the
questions raised by Denmark (EC-85/At.41 and EC-85/At.32, respectively), regarding animal feeding
studies, my understanding is that the applicant has provided sufficient relevant information.
According to the Codex Alimentarius Guideline fo r the Conduct of Food Safety Assessment of Foods
Derived from Recombinant-DNA Plants (CAC/GL 45-200 3, paragraph 53), additional animal testing
will be required if the composition is not comparable to conventional foods, which is not the case of
Monsanto's event corn GA 21.
Question 40bis: Given the information before the Panel, including the first whole food study
and agronomic performance tests (attached, cover letter provided in EC 78/85/At. 19), was a
second animal whole food study requested by Denmark, Austria and Italy (EC-78/85 /At. 67, 40
and 72) necessary or useful to identify potential adverse effects that had not been previously
identified?
Dr. Nutti
672. Answer 40 bis: Based on 78/85/At. 19 and the request by Denmark, Austria and Italy (EC-
78/85 /At. 67, 40 and 72, asking for a second animal whole feeding study, my opinion is that such a
request was not necessary to ensure that conclu sions of the safety assessment were valid. Based
on the Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of Foods Derived
from Recombinant-DNA Plants (CAC/GL 45-2003), paragraphs 53, my understanding is that
additional animal feeding studies may be warranted for GM foods if changes in the biovailability of
the nutrients are expected or if the composition of the GM food is not comparable to conventional
food, and this was not the case of maize GA21.
Pioneer/Dupont high-oleic soybean (260-05)
C/NL/98/09 (EC chronology 87)
Question 41: Given the information before the Panel, including the notification (EC-87/At.1),
additional letters from Monsanto providing additional information (EC-87/At.3, 7, CBI part in
8, and 14) and the conclusion of the Commission on Genetic Modification that this product did
not present environmental risks (EC-87/At.8-9), wa s additional informatio n regarding data on
the composition of high-oleic soybeans and the alteration in the protein profile of this product
requested by the lead CA (EC-87/At.11, 13, and 15) necessary to ensure that conclusions of the
safety assessment were valid?
Dr. Nutti
673. Based on the notification (EC-87/At.1), the additional information regarding data on the
composition of high-oleic soybeans (lectin content and composition data from at least two seasons.)
and the quantification of the change in the protein profile of this product requested by the lead CA
(EC-87/At.11 and 12) were necessary to ensure that conclusions of the safety assessment were valid.
674. It is important to point out that the lead CA has recognized that the soya oil with the changed
fatty acid composition is mainly intended for human use, and not for livestock feed. It is therefore not
expected that these changes will have an impact on livestock feed safety.

Dr. Squire
675. Response on environmental risk. This product is intended for import for processing, not for
growing as a crop. Its potential for environmental risk – compared to other soybean – is through any
difference in the waste the processing generates or any difference in the rate of establishment as a
casual or weedy plant. It would seem that the di fferences in chemical composition of the high-oleic
soybean are unlikely to generate a different type of waste after processing.
676. The company indicate the product does not pose a risk through becoming established as a
plant "since the transgenic soybeans do not differ in their characteristics regarding survival,
multiplication and dissemination from commodity soybeans" (paragraph 42 in EC-87 At-1). While
there are no data presented explicitly on this poi nt, the assertion is nevertheless well founded. In
Europe, the soya-bean (soybean) Glycine max (L.), ex ists as an uncommon casual plant, mostly on
tips and waste ground, originating from imported oil-seed (spilled or dumped), food-refuse and bird-
seed. It is very much less common than plants arisi ng from, say, oilseed rape: in the UK, for instance,
casual soybean was recorded at less than 50 localities over many years in a major compendium of
alien plants (see reference below). It does not spre ad, is not an agricultural weed and will not cross-
pollinate with local plants. It is possible that im ported high oleic soybean would come to exist as a
casual plant in Europe, but there is no reason to in dicate that it would exist in greater numbers, for
longer or at more locations than other soybean. The high-oleic soybean does not have properties that
would cause it to differ markedly in nutritional or toxic properties, and so affect insects or other wild
animals, and the evidence and arguments presented about the extremely low risks regarding horizontal
gene transfer (e.g. to microorganisms) appear correct. Therefore the conclusion of the Commission of
Genetic Modification that the product did not present environmental risk is sound. The requests for
further information (EC-87, At 11, 13, 15) appear to be mainly about safety as a feed, so the
additional year's field data (requested) would not provide substantive new information on the plant's
environmental risk.
Clement EL, Foster MC (1994) Alien plants of the British Isles. Botanical Society of the
British Isles, London.
Monsanto/Syngenta Roundup Ready sugar beet
C/BE/99/01 (EC chronology 88)
Question 42: Given the information before the Panel, including the notification (EC-88/At.1)
and additional information provided by the notifier (EC-88/At.10-11), was the information
regarding allergenicity, molecular characterisa tion, and gene transfer in digestive tracts
requested by the lead CA (EC-88/At.12) necessary to ensure that conclusions of the safety
assessment were valid?
Dr. Nutti
677. Based on the notification (EC-88 At.1) and additi onal information provided by the notifier
(EC-88 At.11-12), my understanding is:
678. EC- 88 At.11_SCI, pg 7, Question 3.1, asks more information on allergenicity. The additional
answer is provided at Section 4, pg 13 of EC-88 At 11, with a very detailed explanation on the
allergenicity assessment. The notifiers provided ag ain the references explaining that there are two
common characteristics snared by most allergens: first, food allergens are generally abundant in food,
usually greater than 1% of the total protein and, seco nd, food allergens are stable to gastric digestion,
and a specific test for gastric stability has been established and validated using a broad array of known

food allergens. Assessment of this two characteris tics along with careful review of the history of
exposure and search for possible sequence similar ity between the GM protein and known allergens
can help minimize the risk of transferring a known allergen, one that is possibly going to be cross-
reactive to a known allergen, or one that could have a high likelihood of being a food allergen. On pg
13 and 14, the notifiers explained the rationale for the test carried out, the decision three used for the
allergenic potential proteins encoded by genes tran sferred to genetically modified crops, derived from
FAO/WHO 1996, ILSI 1996. Although this references and this process were carried out in 1998, it is
very important to point out that the allergenicity assessment was carried out according to the rules that
would be established by the Codex Alimentarius Guidelines for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DN A Plants (CAC/GL 45-2003) and its Annex on
Possible Allergenicity Assessment My understanding in this case is that the necessary information
was provided by the notifiers in the notification, and the additional information, although was not
needed, provided clarification to the lead CA, to the understanding of the allergenicity assessment.
679. EC-88 At.11_SCI, pg 7, Question 3.2: "Can the notifying firms show that the concentration
of GUS protein in the gastrointestinal tract that appears in people and animals after ingesting food
products is sufficient protection against any toxici ty caused by the deglucuronidation of endogenous
or exogenous products?", is properly answered by the notifiers, when they explain that since sugar is
derived from sugar beet and there is very little if any residual protein, there would be no meaningful
exposure to GUS protein with the consumption of refi ned sugar. In my opinion this information was
already in the notification, the additional answer was just an explanation to the lead CA.
680. EC-88 At.11_SCI, pg 7-9, Question 3.3: "On page 64, proteins wich are toxic via acute
mechanisms needs justification. The same is true of the acute toxicity test". With this question, the
lead CA asks for justification for the acute gavage study tests done with mice (provided in the
notification) and information why this tests has been carried out with proteins that are toxic. The
notifier answered the question, explaining that acute gavage tests are usually performed with proteins
that are toxic via acute mechanisms, provided th e reference (EPA and Slojab at al, 1992), and
explained the rationale for using this test with CP4EPSPS proteins. The notifier also reminded that
those proteins were rapid degraded in in vitro digestibility studies, providing more information with
new versions of paragraphs C.2.a (ii), C.2.b (ii), C.2.c (iii) in the Application, in the Appendix of
Section 4 of the document. My conclusion, as far as the toxicity of the protein is concerned, is that
the notifier provided the information requested and l ogical justification why those tests were carried
out, so the information provided at EC- 80 At 11 complemented and explained the data submitted in
the notification ( At 1), clarifying the lead CA, but it is important to point out that the necessary
information on the toxicity of the protein has been presented at the notification, so further information
was not necessary to ensure the conclusions of the safety assessment in this topic (toxicity of the
protein), but it helped very much to understand a ll the issue on toxicity, as the answer was detailed
and explained not only the tests which were car ried out, but also the specific target tissue,
measurements of weight and f ood consumption, low toxicity of the GUS protein, and minor
pathological changes observed at the necropsy, so with the answer provided at AT.11, my
understanding is that no more information was needed in this topic.
Question 43: Given the information before the Panel, including the notification and additional
information provided by the notifier (EC-88/At.10, 11, 13, 14, 15, 16, and 18-26), was the
information regarding molecular characterisation and allergenicity of event '77' requested by
the lead CA (EC-88/At.27-28) necessary to ensure that conclusions of the safety assessment were
valid?

Dr. Nutti
681. The information required by the lead CA (EC-88 At. 27-28) for allergenicity was not
necessary to ensure that the conclusions of the safety assessment were valid. The previous information
on this item covered all the points recommended by WHO/FAO Expert Consultation 1996, and the
Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of Foods Derived from
Recombinant-DNA Plants (CAC/GL 45-2003) and its Annex on Possible Allergenicity Assessment.
Monsanto MaisGard Roundup Ready (MON 810 & GA21) corn (stack)
C/ES/99/02 (EC chronology 94)

Question 44: Given the information before the Pane l, including the notification (EC-94/At.1-3),
was the information requested by the Neth erlands(EC-94/At.12) concerning molecular
characterization, DNA sequence analysis of the insert ion event, analysis of protein levels, effect
of glyphosphate treatment, composition, toxico logy and the request for a study on dairy cows
necessary to ensure that conclusions of the safety assessment were valid?
Dr. Nutti
682. Concerning the information requested by Netherlands in EC-94/At.12, I will answer the
questions related to my background.
683. Question 3 – This information is not necessary to ensure the validity of the safety assessment.
The lead CA was asking for more explanation about the higher level of m EPSPS protein in MON
810xGA21 maize line, as the Committee did not accept th at they are due to biological variety. My
opinion is that the explanation given by the notifier was acceptable, since in EC-94/ At 13, the notifier
presents the results of second year study in order to test this assumption, and the results confirmed
that the expression levels were similar.
684. Question 4 – the Committee asked to be provided data to confirm that glyfosate treatment has
no effect on the composition of maize plants used and justification for the statistical analysis
presented at annex 2 of the dossier. It was also poi nted out that regarding question 3, the glyfosate
treatment was not considered to be a possible factor that might influence the level of expression of the
introduced trait. My understanding is that for the comparison of a GM with a non GM product,
usually the treatment with herbicide is not required, as the tests are done in order to compare the effect
of the new trait, and not the effect of the herbicide. In this case, the applicants had provided data with
glyfosate treated and non-treated product, so the in formation asked was not necessary to ensure the
safety assessment. This was confirmed at the answer provided by the applicant at EC-94/ At 13,
where references were provided that the treatment with glyfosate does not affect the composition of
GM maize or GM soybean.
685. Question 5 – the Committee recognized that data was provided on the comparison of
MON810x GA21 and the parental lines MON 810 a nd GA 21 for grain and forage, and asked for
more information, including a comparison of the new hybrid with non-transgenic parental lines, as the
comparison presented was with the transgenic parental lines. The Committee asked for information on
the composition of ferulic acid, p-commaric acid, raffinose and inositol, and in this case the
Committee accepts the comparison between the new hybr id with the parental transgenic lines. My
understanding is that there are different approaches that can be used when GM traits have been
combined by traditional breeding (stacked), one is that the comparison of the new hybrid should be
done with the GM parental lines, and this GM parental lines should have been compared with
conventional counterpart; another is that the new hybrid should be compared with non-transgenic

parental lines. The problem with the second option is that sometimes the relevant parental lines may
not been developed or may exist only in a genetic background which is not comparable to the stacked
hybrid.
686. The applicant has chosen the first option for th e comparison, as data was presented with the
information of the equivalence of GA 21 and MON 810 to their nearest non-transgenic isolines, and
also the new hybrid MON810xMONGA21 with the pare ntal transgenic lines. My understanding is
that the information submitted by the applicant wa s sufficient to ensure the validity of the safety
assessment. As far as information on the composition of ferulic acid, p-commaric acid, raffinose and
inositol, were concerned, my understanding is that this supplementary information was not necessary,
as the applicant had already submitted information indicating that none of the corresponding pathways
have been altered by the breeding of new traits. The applicant also informed that Phytic acid has been
considered an important seconda ry metabolite in maize and its measurement encompasses a
biologically relevant form of inositol.
687. Question 6 – toxicological tests, the Committee asked for a semi chronic feeding study on
mice with grain or meal from MON 810x GA21maize. Based on Codex Alimentarius Guidelines for
the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants (CAC/GL
45-2003), paragraphs 34 to 43, Assessment of Possibl e Toxicity, we have at para 37 that the use of
appropriate conventional toxicology or other studies on the new substance may be necessary, if taking
into account its function and exposure, there are doubt s on the safety of the new substance. In this
case the applicant has provided in the dossier (EC-94/At.1-3), all th e information usually requested
for the food safety assessment, for toxicological ass essment he presented safety studies on protein
digestion in mammalian gastric and intestinal sy stems, acute gavage studies in mice, homology to
known toxins and allergens and exposure to human diet. Based on Codex Alimentarius Guidelines for
the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants (CAC/GL
45-2003), paragraphs 53, my unde rstanding is that additional animal feeding studies may be
warranted for GM foods if changes in the biovail ability of the nutrients are expected or if the
composition of the GM food is not comparable to c onventional food, that is not the case of the new
hybrid MON 810x GA21 maize. My understanding is th at the information submitted by the applicant
was sufficient to ensure the validity of the safety assessment.
Dr. Andow
Necessity to ensure that conclusions were valid:
688. I will answer this question with respect to the effect of glyphosphate treatment and
toxicology.
689. The relevant question from the Netherlands CA (E C-94/At.12, 17 July 2000) on the effect of
the glyphosate treatment is reproduced below.
4. The field trials and block treatment . In Appendix II of the dossier, the
compositional analyses of the MON 810 x GA21 maize are described. The following
comparisons are made: components of MON 810 x GA21 maize/forage with
components of MON 810 maize/forage (both maize lines not treated with
Glyphosate) and components of MON 810 x GA21 maize/forage with components of
GA21 maize/forage (both maize lines not treated with glyphosate). Then, the
following statement is made: "the significant differences in component values are
only relevant where the mean value of the MON 810 x GA21 maize line was either
greater than or less than both parental lines". However, this assumption is only viable

if there is no effect of Glyphosate treatme nt on the composition of the maize lines
since only half of the maize plants used for analyses (maize lines MON 810 x GA21
and GA21) are treated with Glyphosate. Therefore, the Committee would like to be
provided with data to confirm that Gl yphosate treatment has no effect on the
composition of the maize plants used and to justify the statistical analyses and
comparisons between MON 810 x GA21 maize/forage (not treated with glyphosate)
and MON 810 x GA21 maize/forage (treated with glyphosate).
In relation to question 3: mEPSPS protein levels in MON 810 x GA21 maize line are
compared with the levels of mEPSPS protein in the parental maize lines MON 810
and GA21. In these comparisons, the Glyphosate treatment was not considered to be
a possible factor that might influence the expression levels of the introduced traits.
690. The relevant question from the Netherlands CA (E C-94/At.12, 17 July 2000) on toxicology is
reproduced below.
6. The toxicological tests. Recently, it was discovered that in genetically
modified soybeans parts of the vector used for the modification were scattered over
the genome of the soybeans. It can not be excluded that these parts are able to form
proteins. The Committee is of the opinion that other plants that are modified with the
particle acceleration method also might contain some unknown parts of DNA. The
presence and safety of these DNA parts, howev er, can not be guaranteed with the
current molecular biological techniques. Therefore, the Committee finds that, beside
the molecular biological characterization of the modified genome, a semi-chronic
toxicity study with relevant parts or prod ucts of the plants are necessary to assess the
safety of the modified plant for the consum ers. In the case of the new maize hybrid
MON 810 x GA21, this would mean, for example, a semi-chronic feeding study on
mice with maize grain or meal from the MON 810 x GA21 maize plant.
691. The statistical analysis used is given in Appendix II, page 90 of EC-94/At.2, 29 February
2000.
Statistical analyses of the U.S. and E.U. composition data was conducted separately
and combined using a mixed model analysis of variance:
Yijk = U + Li + Bij + Tk + eijk,
where U = overall mean, Li = random location effect, Bij = random spray block within
location effect, T k = treatment line effect, and e ijk = residual error. The component
values for the RR/YG lines were compared to those of the corresponding single trait
parental lines (RR and YG) to determine significant differences. SAS ® software was
used to generate all summary statistics a nd perform all analyses (SAS Institute, 1989,
1990, 1996). Report tables present p-values from SAS ® as either <0.001 or the actual
value truncated to three decimal places.
692. The experimental design for both the US and EU field studies was similar. There were six
sites in the US trial and seven sites in the EU tria l. At each site the field was split into two glyphosate
treatments (plus glyphosate or no glyphosate). All of the RoundUp Ready ® (RR) varieties were
planted in the plus glyphosate whole plot. Those lacking RR were in the no glyphosate whole plot.
Within each whole plot, the relevant YieldGard
® (YG) varieties (both DeKa lb and Holden parentage)

and those lacking YG were planted (presumably randomly) as the split plots within the whole plots.
Thus there were four split treatments within each of the two whole plots.
693. From the description in paragraph 691, it would appear that the notifier compared RR+ versus
RR- means based on estimated standard errors of the means from the stated statistical analysis.
694. In my opinion, the notifier used the wrong statis tical analysis. The experiment is set up as a
randomized complete block-split plot design with the following statistical model:

695. where i indicates location, j the glyphosate treatment and presence/absence of the RR gene,
and k a treatment level. Y is the response, µ is the grand mean, RR is the glyphosate treatment/RR
gene effect, εij is the whole plot error for testing the RR effect, T is the treatment effect, nested within
split plots, and εik(j) is the residual error. The T effect can be decomposed into the following
orthogonal effects:
T = YG + V + YG * V + YG * RR + V * RR + YG * V * RR
696. where YG is the effect of the YG gene, V is the effect of parental type (DeKalb versus
Holden), and the other terms are the interaction effects. All of the T components have 1 df. As there
is no true replication within locations, it is essential to treat locations as replicates or there would be
no source of error variation. In doing this, it is readily recognized that locations are identical to
blocks, so the whole plot design is a randomized complete block design. The appropriate ANOVA
table for this analysis for the EU trial is:
LEVEL EFFECT df MS F p
Level 1 Location (L) 6
 RR 1
 Error 1 6
Level 2 T 6
 YG 1
 V 1
 YG*V 1
 YG*RR 1
 V*RR 1
 YG*V*RR 1
 Error 2 36

697. This analysis would allow direct statistical test of all relevant effects. It should be noted,
however that the RR gene effect is totally confounded with the effect of the application of glyphosate
in the experimental design. Thus it is not possible to estimate an RR effect independent of a
glyphosate effect.
698. Moreover, the notifier treats each of the many response variables as if they were completely
independent responses. Statistically it is inadvi sable to treat each response independently with a
series of ANOVAs (which is what the notifier did). In actuality it is likely that many of the response
variables will be correlated with each other. Thus, the notifier should have considered several

possible multivariate responses, and conducted sever al intelligently planned MANOVAs prior to the
many ANOVAs. Statistically it is inadvisable to use all of the data in a single large MANOVA.
699. Thus the CA was justified to request additi onal information on the glyphosate treatment, the
statistical analysis, and the comparisons made in Appendix II. These were necessary to ensure that
conclusions of the safety assessment were valid.
700. With respect to the toxicity question, the NL-CA is correct to suggest that there may be parts
of the vector scattered over the genome. However, with respect to maize, it should be possible to
eliminate most of these via the backcrossing methods used to transfer a transgene into a
commercializable inbred line. The only ones that are likely to remain are ones that are closely linked
to the transgene locus. It would seem that the notifier would have other alternatives to the toxicity test
to address the concern of the Netherlands CA. By providing details such as the number of backcross
generations, it would be possible to calculate a proba bility that vector parts remain scattered around
the genome. Soybean, being predominantly selfing would not readily eliminate such vector parts
from its genome during the plant breeding process. T hus, while the concern is valid and needs to be
addressed to ensure that the conclusions of the safety assessment are valid, the request for toxicity
testing is not necessary to ensure that the conclusions of the safety assessment were valid for maize.
701. The notifier submitted the original notification to the NL-CA on 29 February 2000. The NL-
CA requested additional information on 17 July 2000. A 139 day interval elapsed. This seems to be a
little longer than necessary to complete the review and request additional information.
Question 44bis: Given the information before the Panel, including the first whole food study
and agronomic performance tests (EC 94/At.3, pp. 40), was a second animal whole food study
(EC 94/At.12) necessary or useful to identify potential adverse effects, including those associated
with small random DNA insertions, that had not been previously identified?
Dr. Nutti
702. Answer 44bis: Based on (EC 94/At.3, pp. 40), and the request by the lead CA (EC
94/At.12), asking for a second animal whole feedi ng study in order to identify potential adverse
effects, my opinion is that such a request was not necessary to ensure that conclusions of the
safety assessment were valid.
703. Based on the Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of
Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003), paragraphs 53, my understanding
is that additional animal feeding studies may be warranted for GM foods if changes in the
biovailability of the nutrients are expected or if the composition of the GM food is not comparable to
conventional food, and this was not the case of maize MON 810 & GA21.
Question 44ter: In the context of this corn product which was produced through conventional
breeding from biotech parents, is data regarding the safety of the biotech parent plants relevant
to the safety assessment of the hybrid plant pro ducts? Given the results of the risk assessment
studies of the biotech parents, were additional studies necessary or useful to ensure that the
conclusions of the safety assessment related to the hybrid plants were valid?
Dr. Nutti
704. Answer 44ter: As I already pointed out in my answer of question 44, my understanding is
that there are different approaches that can be used when GM traits have been combined by traditional

breeding (stacked), one is that the comparison of the new hybrid should be done with the GM parental
lines, and this GM parental lines should have been compared with conventi onal counterpart; another
is that the new hybrid should be compared with non-transgenic parental lines. The problem with the
second option is that sometimes the relevant parent al lines may not been developed or may exist only
in a genetic background which is not comparable to the stacked hybrid. The applicant has chosen the
first option for the comparison, as data was presented with the information of the equivalence of GA
21 and MON 810 to their nearest non-transgen ic isolines, and also the new hybrid
MON810xMONGA21 with the parental transgenic lines. My understanding is that the
information submitted by the applicant was sufficient to ensure the validity of the safety
assessment. In this case I believe that given the results of the risk assessment studies of the biotech
parents, no additional studies were necessary or useful to ensure that the conclusions of the safety
assessment related to the hybrid plants were valid.
Novel foods
Monsanto Roundup Ready corn (GA 21)
(EC chronology 91)

Question 45: Given the information before the Pa nel, were there scientific or technical reasons
which explain why the process of validating the detection method took at least fourteen months
(June 2002-August 2003)? (See EC-91/At.49-56)?
Dr. Nutti
705. Although detection methods are not my main area of expertise, I had analysed the information
presented at EC-91/At.49-56, and I noticed that at EC-91/At.50, the applicant submits not only the
protocol ( methodology) used for the detection, but also comments on the main issues that should be
taken into consideration, as the preparation of standard DNA solution, validation parameters,
precision of measurements, accuracy, robustness, range and sensitivity.
706. EC-91/At.51, the applicant informs what are the materials that should be used in the analysis.
707. EC-91/At.52, the applicant submits more speci fic information on the protocol already sent
and specific data for the validation of GA21 detection methodology.
708. EC-91/At.53, the applicant informs that the reagents will be sent.
709. EC-91/At.54, the applicant apologizes for the delay and informs that more reagents will be
sent. Applicant asks information for the amount and type of reagent requested.
710. EC-91/At.55, JRC extends the deadline and answers the applicant about the amount and type
of reagents.
711. EC-91/At.56, JRC sends the draft protocol.
712. Based on the information available, I have th e impression, that there were no scientific or
technical reasons which explain why the process of validating the detection method took at least
fourteen months. It is important to point out th at the existing method submitted by the applicant at
EC-91/At.50, was complete and detailed, being alr eady in use, so the JRC was supposed to test and
validate it.

Syngenta Bt-11 sweet corn
(EC chronology 92)

Question 46: Were detection methods commercially available in 2002 sufficient to enable the
detection of the transgenic proteins expressed by the plant line Bt11 sweet corn?
Dr. Nutti
713. The methods available in 2002 were sufficient to enable to detect the DNA and the protein of
Bt11sweet corn.
714. Beginning in the 1990's, commercial protein-b ased and DNA-based detection methods have
become available. Both methods can be used in a qualitative and a quantitative way. Early DNA-
based methods have been mainly designed as qualitative methods only. More recently, there has been
a shift towards quantitative PCR methods. Today, most published methods are based on real-time
PCR technology that can be operated both in a qualitative and a quantitative way. Some DNA-based
methods were designed for the identification of a specific event while others were designed to identify
only a certain vector construct or a common gene tic element (e.g., 35S promoter). Today, the
development of these three different types PCR met hods continues. The selection of method depends
on the intended purpose. The experience for protein- based methods is similar to the experience with
DNA-based methods – there are many commercially available protein-based methods also. These
methods can confirm the presence or absence of spec ific proteins expressed by the different biotech
events. These protein-based methods can be applied in both a quantitative and qualitative fashion. The
protein based methods, while not event-specific, can identify uniquely most commercial events.
Dr. Healy
715. Response: In 1998, the SCP expressed a favourable opinion about the safety and human
health impacts of the field maize line Bt-11. Subsequently the European Commission published its
decision to grant consent to the marketing of the Bt-11 line of maize, but not the cultivation of this
line. In November 1998, Novartis submitted to the Dutch authorities a request for an authorisation of
food products derived from fresh and processed sweet corn carrying the Bt-11 transformation event.
This request was subsequently sent to the Eur opean Commission and dist ributed to the Member
States, some of which raised issues of concern. The Scientific Committee for Food (SCF) reviewed
the scientific information underpinning the request for authorisation and expressed the view that 'Bt-
11 sweet maize is as safe for human food use as its conventional counterparts'.
Background:
716. The Bt-11 corn line contains a single copy of two transgenes, namely the cry1A(b) and pat
genes. The cry1A(b) gene encodes one of the Bt toxins derived from the common bacterium Bacillus
thuringiensis. The toxins are selectively active against groups of insects, particularly moths and
butterflies, beetles, flies and mosquitos. The Cry1A(b) protein toxin is selectively active against
lepidopterans (ie moths and butterflies). The pat gene is derived from a common soil bacterium and
encodes the enzyme phosphinothricin acetyl transferase (PAT), which inactivates the herbicide
glufosinate ammonium and confers herbicide tolerance to the genetically modified corn.
717. No other marker genes, including antibiotic resistance genes were transferred, to Bt-11 corn.

Detection methods:
718. Methods to detect transgene(s) and their produc t(s) may be based on either the detection of
specific DNA sequences (Polymersase Chain Reaction; PRC) or on the identification of the proteins
produced by the transgene(s). The documents subm itted by Novaris/Syngenta to support the approval
of products derived from Bt-11 reference several D NA based detection methods for Bt-11 corn lines.
These include the method of Zimmermann et al (2 000), based on quantitative polymerase PCR at the
integration site of the transgene, a PCR based me thod subjected to pre-validation studies by the Joint
Research Centre (EC 92- 59; it is not clear from the available documents if this method is also based
on the method of Zimmermann et al, 2000) and another method developed by Syngenta in
conjunction with the Joint Research Centre and s ubjected to validation procedures (EC 92-60, 63, 64,
65 and 66; Rolling et al 2003). However, it app ears that none of these DNA based procedures were
available commercially in 2002 (Codex Inter governmental Taskforce on Foods Derived from
Biotechnology, 2002; Bonfini et al, 2002).
719. Methods for the detection of the transgenic pr oteins expressed by Bt-11, as raised in question
46, could focus on either of the PAT or the Cry1A(b) proteins, as the genes encoding both proteins
have been transferred to Bt-11 and both are expres sed in Bt-11 plants. The availability of detection
methods for the detection of the PAT protein are discussed under question 17; the discussion here
focuses on the detection of the Cyr1A(b) protein.
720. A method based on the detection of the Cry1A(b) protein by an antibody was available in
2002 (Intergovernmental Taskforce on Foods Derive d from Biotechnology, 2002). This method had
been developed to quantitatively determine the leve ls of Cry1A(b) in the transgenic corn line
MON810. The method was subjected to a large inter-laboratory validation trial to test the
performance of a kit based on the method and used co rn flour samples as the test material (Bonfini et
al, 2002).
721. While the test method was designed to quantify Cry1A(b) in MON810 plants and their
derivative food products, the method should be able to detect Cry1A(b) in any plant variety in which
this protein is expressed in sufficiently high levels . It has been demonstrated that the method does
indeed detect the Cry1A(b) from Bt-11 in samples containing as little as 0.1% Cry1A(b) (Stave,
2002).
722. It should be noted that the protein based detection method for Bt-11 does not specifically
detect Bt-11 plants and their food products. Rather it detects any plant and food product that contains
Cry1A(b) protein at detectable levels – a number of crops have been genetically modified with the
gene encoding Cry1A(b). This is a general li mitation of protein based detection methods.
Furthermore, the method is only functional for tho se foods derived from plant tissue that express the
protein and accurate quantification is not always possible (Stave, 2002).
723. It is also notable that both DNA and protein based methods are effective in foods only if the
DNA or protein is retained intact through an y processing (eg DNA and proteins are generally
removed in oil extraction and refinement processes ) and can be extracted and purified from food
matrices.

Conclusion:
724. The available evidence indicates that an immunoassay based detection method suitable for the
detection of Cry1A(b) protein in Bt-11 trangenic plants was commercially available in 2002.113

113 References Dr. Healy:

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included in Standard A18/Standard 1.5.2 – Food Produced Using Gene Technology.
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Question 47: Given the information before the Pa nel, were there scientific or technical reasons
which explain why the process of validating the detection method took twelve months (EC-
92/At.54-56, 57-65, and 66)? What would constitute an adequate amount of material to be used
in the detection method validation? What are adequate performance indicators for pre-
validation results for this product?
Dr. Nutti
725. Although detection methods are not my main area of expertise, I had analysed the information
presented at EC-92/At.54-56, 57-65, and 66 and I no ticed that the applicant submitted the material
and proposed methodology, JRC tested the method and at EC-92/At.56, JRC points out some
problems that were found , mainly with the am ount of DNA detected in some samples , asks for
comments and more material for testing.
726. EC-92/At.57, JRC explains to the applicant the main issues discussed by phone and
acknowledge the reception of the material for re-starting the validation of the tests.
727. EC-92/At.58, the applicant submits more information on the methodology.
728. EC-92/At.59, confidential report from JRC, informing that the method submitted by the
applicant could not be accepted for a full validation before a final method optimization is carried out,
pointing out problems on the amplification of the GM specific system. JRC informs that they are
willing to discuss the problems with the applicant and share expertise.

USA Food and Drug Administration (1997). Consultation Procedures under FDA's 1992 Statement of
Policy: Foods Derived from New Plant Varieties. www.cfsan.fda.gov/~lrd/consulpr.html.
USA Food and Drug Administration (2001). Proposed Rule: Premarket Notice Concerning
Bioengineered Foods Premarket Notice Concerning Bioengineered Foods.
www.cfsan.fda.gov/~lrd/fr010118.html.
Van Haver E., de Schrijver, A., Devos, Y., Lievens, S., Renckens, S., Moens, W. (2003). Guidance
Notes for the Safety Assessment of Gene tically Modified Crops for Food and Feed Use. Report of the Belgian
Biosafety Advisory Council. Scientific Institute of Public Health.
www.biosafety.be/NF/GuidanceNotes/Documents/SafetyAssessmentGMFoodFeed.
Windels, PI., Taverniers, A. Depicker, E. Van Bock staele, and M. De Loose. (2001). Characterization
of the Roundup Ready soybean insert. Eur Food Res Technol. 213:107-112.
World Health Organisation (1987). Principles fo r the Safety Assessment of Food Additives and
Contaminants in Food. Environmental Health Criteria 70. World Health Organisation, Geneva.
World Health Organisation (1993). Health Aspects of Marker Genes in Genetically Modified Plants.
Report of a WHO Expert Consultation on Foods Derived from Biotechnology. World Health Organisation,
Geneva.
World Health Organisation (2000). Safety Aspects of Genetically Modified Foods of Plant Origin.
Report of the Joint FAO/WHO Expert Consultation on Foods Derived from Biotechnology. World Health
Organisation, Geneva.
World Health Organisation (2001a). Evaluation of the Allergenicity of Genetically Modified Foods.
Report of the Joint FAO/WHO Expert Consultation on Foods Derived from Biotechnology. World Health
Organisation, Geneva.
World Health Organisation (2001b). Safety Assessment of Foods Derived from Genetically Modified
Micro-organisms. Report of the Joint FAO/WHO Expert Consultation on Foods Derived from Biotechnology.
World Health Organisation, Geneva.
World Health Organisation (2003). Safety Assessmen t of Foods Derived from Genetically Modified
Animals, including Fish. Report of the Joint FAO/WHO Expert Consultation on Foods Derived from
Biotechnology. World Health Organisation, Geneva.
Zimmermann, A., Luthy, J, Pauli,U. (2000). Event Specific transgene Detection in Bt-11 Corn by
Quantitative PCR at the Integration Site. Lebensm.-Wiss.u.-Technol. 33: 210-216.

729. EC-92/At.61-65, several e-mails are exchanged between the applicant and JRC, about the
methodology validation.
730. EC-92/At.66, JRC informs the conclusion of the validation and acceptance of the method.
731. Based on the information available, I have th e impression, that there were no scientific or
technical reasons which explain why the process of validating the detection method took twelve
months.
732. I don't have expertise or references to answer about the adequate amount of material to be
used in the detection method validation and adequate performance indicators for pre-validation results
for this product.
Bayer LibertyLink soybeans
(EC chronology 93)

Question 48: Given the information before the Panel, what type of data should be used to
evaluate substantial equivalence of these soybeans given their intended end-use (human
consumption)? Was information regarding substantial equivalence, presence of PAT
 DNA and
pat protein requested by the Belgian Biosafety Council (EC-93/At.11) necessary to ensure that
conclusions of the safety assessment were valid?
Dr. Nutti
733. Based on the information provided by the applicant (EC-093/At. 01-2) and the request for
additional information by the Belgian Biosafet y Council (EC-93/At.11), as far as substantial
equivalence is concerned, my understanding is that although the notifier provided a good comparative
analysis of two transgenic lines and one non-transgen ic line, with information on a large number of
nutrients and anti-nutrients, the request for data on composition (soluble and insoluble nutritional
fibres, vitamin, mineral and trace elements contents) was proper since this information is relevant for
the comparison. This is due to the fact that, when a product is a source of fibre, as in this case, the
comparison should take into account its soluble a nd insoluble fibres contents. Besides, products
derived from soybean are considered to be a good source of vitamins and minerals.
734. The request for the company to present information on the presence of PAT DNA and PAT
protein in derived soya products is also correct , as the notifier presented only data on the protein
content in the seeds and derived products, but did not present any specific information on PAT
protein.
735. My understanding is that the requests were nece ssary to ensure that the conclusions of the
safety assessment were valid.
Question 49: Given the information before the Pa nel, including the application (EC-93/At.1-2),
was additional information regarding nutritional and biochemical characterization and toxicity
of the transgenic plant requested by the G reek and Italian authorities (EC-93/At.16-17)
necessary to ensure that conclusions of the safety assessment were valid?
Dr. Nutti
736. Based on the application (EC-93/At.1-2), the request from the Greek authorities (EC-
93/At.16), items a and b, regarding to the information on the analytical methods used for

determination of anti-nutrients and data on th e nutritional and composition parameters of the
transgenic and non-transgenic products, is correct, as I could not find this necessary information in the
dossier. Based on the application (EC-93/At.1-2), concerning the request from the Italian authorities
(EC-93/At.17), my understanding is:
737. Questions 1 and 2, related to the herbicide treat ment in the tests and possible residues of this
treatment in the final product. My understanding is that the risk assessment conducted here is related
to the GMO and not to the herbicide, so this info rmation is not relevant fo r the conclusions of the
safety assessment. It is important to point out that the herbicide residue in the product has to be
within the limits established by JEFCA and the Codex Alimentarius, so the herbicide has been
assessed in a different study. Sometimes, companies that are carrying out the safety assessment of
herbicide resistant GMOs, perform fi eld tests with and without herbicides, but this is not the main
objective of the evaluation, as we need to compare the transgenic and non transgenic crops and not the
utilization of the herbicide.
738. Question 3, asking detailed information on th e analytical methods used to evaluate
composition in macro and micro nutrients and the cont ent of anti-nutrironal factors, is correct, as I
could not find this necessary information in the dossier.
739. Question 4, about accreditation status of the laboratory that carried out the composition
analysis, is correct, as according to the Codex Guidelines the analysis should be performed under
Good Laboratory Practices.
740. Question 5, asking the study of the acute toxic ity of the PAT protein to be presented, is
correct, as I could not find this necessary information in the dossier.
741. Question 7, asking data on substantial equivalence to be presented on composition or animal
tests carried out by the company, is correct, as I could not find this necessary information in the
dossier.
742. Remark on the necessity of carrying out a study of sub-acute toxicity to obtain a more
detailed assessment of toxicological aspects. In my opinion, if the information on the equivalence
were properly supplied and an acute toxicity gavage study was carried out with satisfactory result, it
would be not necessary to carry out a sub-acute test. As the notification does not present all the
necessary information on equivalence, I cannot judge this request.
Question 50: Given the information before the Panel, including the application and the
information provided by Aventis Crop Science (EC-93/At.21), was additional information
regarding nutritional composition requested by Belgium (EC-93/At.23) necessary to ensure that
conclusions of the safety assessment were valid? Does any scientific evidence exist to
demonstrate that genetically-modified soybean varieties are toxic to humans or animals? If so,
what risk management options exist to mitigate any such risks?
Dr. Nutti
743. Looking at EC-93, At.21 and 22, I understand th at the notifier has provided information with
four additional data packages on nutritional comp osition of Liberty Link Soybean A2704-12 and
A5547-127. I could not find these additional information in the files that were sent to me, as At .21.
At 22 are only the letters containing the additional information.

744. In EC-93/At.23, Belgium asks again for the data on neutral and acid detergent fibre, and as
stated before, I agree that this is information necessary for the comparison among the transgenic and
non-transgenic lines. So, I agree with this request but I cannot find the data packages on nutrition
composition. Therefore, if the notifiers had pr ovided the requested information on fibre, the
compositional comparison could be concluded, but it seems that the lead CA could not find these data
in the additional packages.
745. As far as the broiler chicken study is concerned, at the original dossier, EC-93/At.2. pg 29, it
is described that these studies were conducted at the University of Guelph (Leeson 1997 and 1998),
but I did not find a copy of the study in the dossier or in the supplementary information, so I don´t
know if the lead CA has the study or not. My understanding is that this specific information is
relevant for the risk assessment, and the dossier informs that the results showed no difference,
although they are not presented. The scientific ev idence provided was not sufficient to demonstrate
that the genetic modified soybean is toxic to humans or animals. The problem was that the
information was not complete in order to fulfil all the steps for the risk assessment. So, before
concluding that the product was toxic, the request ed information was necessary to conclude the safety
evaluation.
746. I would not go into risk management options to mitigate risks, if I am not sure if such risks
exist. Therefore, my conclusion is that the lead CA could not conclude about the toxicity of the
product or about the nutritional and compositional equivalence either, so it could not be stated that the
product might be safe or toxic.
Pioneer/Dow AgroSciences Bt corn Cry1F (1507)
(EC chronology 95)
Question 51: Given the information before the Pa nel, including the application (EC-95/At.1-2)
and the responses from Pioneer/Dow AgroScienc es (EC-95/At.10-12), was information
regarding molecular characterization, compositio nal analyses and toxicological analyses of the
product requested by the Health Council of the Netherlands (EC-95/At.8 and 13) necessary to
ensure that conclusions of the safety assessment were valid?
Dr. Nutti
747. Based on the application presented in EC- 95/At.1-2 and the no tifiers' answers in
EC-95/At.10-12, I will comment on the compositional analysis and the toxicological analysis
requested by Netherlands (EC-95/At.8 and 13):
748. Studies for the analysis of nutrient composition of grain were carried out in Chile (1998 and
1999), France and Spain (1999), comparing the GMO a nd the conventional crop, and also the use of
the herbicide, concluding that 1507 maize is compar able and nutritionally equivalent to grain from
existing commercial maize hybrids and spraying with glufosinate ammonium does not have an effect
on the nutrient composition of the 1507 maize grain. The lead CA asked for 3 seasons of field tests in
order to compare the results, but generally 2 seasons are acceptable. My understanding is that the
notifier has provided 2 seasons in Chile and one season in France and Spain. I also understand that the
locations in Chile, Spain and France are a represen tative set of locations. In EC-95/At.10-12, the
notifier explains the seasons and informs about other tests carried out in France, Italy and Bulgaria in
2000, so my understanding is that the notifier has undertaken different representative places for more
than 2 seasons so and, therefore, the additional request was not necessary.

749. The lead CA requested that the compositional data should be complemented with levels of Fe,
Cu, Se, Zn, Vitamin A, vitamin B6 and nicotinic acid. At In EC-95/At.11, pages 10 to 12 , the notifier
supplied the requested data on minerals and Vita min A, vitamin B6 and nicotinic acid. So, my
understanding is that this additional information covered the necessary items for the nutrient
comparison, concluding that the GM and non GM crop were equivalent. My understanding is that the
additional request was necessary and it was covered with the supplementary information.
750. Data on the broiler chicken study was also s upplied in At EC-95/At.11, annex 4 of the
application. So, the additional request was not necessary.
751. The Committee asked the original papers of Brooks (2000), Evans (1998), Glatt(1999)
Herman (2000), Kuhn (1998), Meyer (1999) and Pf ister (1996), which were not included in the
dossier. The lead CA also pointed out the impor tance of the equivalence of the novel proteins
expressed in the 1507 maize and the proteins used in the toxicological tests. A 90-day oral toxicity
study with maize grain or meal on mice was require d in order to provide additional data that no
observed negative effects on food safety would result from a possible unintended change in the 1507
maize. My understanding is that this additional request was necessary and it was covered with the
supplementary information as in EC-95/At.11, the applicant presented all the requested papers, and
information regarding the equivalence of the proteins (page 13), so this information was properly
provided. As far as the 90-day oral toxicity study with maize grains in rats is concerned, the applicant
informed that this study was under way and it would be submitted to the Committee by July 2002.
Furthermore, according to the Codex Alimentarius Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinan t-DNA Plants (CAC/GL 45-2003, paragraph 53),
additional animal testing will be required if the co mposition is not comparable to conventional foods,
which is not the case of Pioneer/Dow AgroSciences Bt corn Cry1F (1507).
Question 52: Given the information before the Panel, including the application and the
responses from Pioneer/Dow AgroSciences (referen ced above), was information regarding the
potential unintended expression of allergenic proteins requested by Gezondheistsraad
(EC-95/At.15) necessary to ensure that conclusions of the safety assessment were valid?
Dr. Nutti
752. My understanding of the request in EC-95/At.15 is that the Committee discussed the weight
of evidence and all information and concluded that sufficient data on the CRY1F protein was
therefore available, substantiating the food safet y of the 1507 maize regarding the presence of this
novel protein. So far, as my knowledge goes, the information submitted for allergenicity was enough
and there was no need for providing additional in formation on putative fusion proteins from the
flanking regions of the insert in 1507 maize.

Monsanto Roundup Ready corn (NK603)
(EC chronology 96)
Question 53: Given the information before the Pa nel, including the application (EC-96/At.1-2),
was information regarding molecular characteriza tion, toxicity effects of unintended changes
and compositional data requested by Gezondheitsraad (EC-96/At.7) necessary to ensure that
conclusions of the safety assessment were valid?
Dr. Nutti
753. Based on the application presented in EC-96/At.1-2, I will comment on the compositional
analysis and toxicological analysis requested by Gezondheitsraad (EC-95/At.7):
754. Question 2, concerning the request for a semi chronic toxicity study in mice or rats, using
maize grain or meal, in order to rule out possible undesired effects of additional, unidentified changes.
Based on the Codex Alimentarius Guidelines for th e Conduct of Food Safety Assessment of Foods
Derived from Recombinant-DNA Plants (CAC/GL 45- 2003), paragraphs 34 to 43, Assessment of
Possible Toxicity, we have at para 37 that the u se of appropriate conventional toxicology or other
studies on the new substance may be necessary, if ta king into account its function and exposure, there
are doubts on the safety of the new substance. In this case the applicant has provided in the dossier
(EC-96/At.1-2) all the information usually requested for the food safety assessment, for toxicological
assessment he presented safety studies on CP4EPS PS protein digestion in mammalian gastric and
intestinal systems, acute gavage studies in mi ce, homology to known toxins and allergens and
exposure to human diet. It was al so confirmed that the GM maize was equivalent in composition and
nutrition to the conventional counterpart. Therefore, in my opinion, there was no need for requesting a
semi chronic toxicity study in mice or rats, using maize grain or meal, in order to rule out possible
undesired effects of additional, unidentified cha nges. Furthermore, according to the Codex
Alimentarius Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants (CAC/GL 45-2003, paragr aph 53), additional animal testing will be
required if the composition is not comparable to conventional foods, which is not the case of
Monsanto Roundup Ready corn (NK603).
755. Question 3 asked clarification on the data of the field trials presented in appendix II-IV of the
application. My understanding is that the inform ation was there and they were just asking for the
exact description of the tests and the statistical design.
Question 53bis: Given the information before the Panel, including the first whole food study
and agronomic performance tests (EC 96/At.2 , pp. 102-103), was a second animal whole food
study (EC 96/At. 7) necessary or useful to iden tify potential adverse effects, including those
associated with small random DNA insertions, that had not been previously identified?
Dr. Nutti
756. Answer 53bis: Based on (EC 96/At.2, pp. 102-103), and the request by the lead CA (EC
96/At. 7) asking for a second animal whole feeding study in order to identify potential adverse effects,
my opinion is that such a request was not necessa ry to ensure that conclusions of the safety
assessment were valid.
757. Based on the Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of
Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003), paragraphs 53, my understanding
is that additional animal feeding studies may be warranted for GM foods if changes in the

biovailability of the nutrients are expected or if the composition of the GM food is not comparable to
conventional food, and this was not the case of maize NK603.
Question 54: Given the information before th e Panel, including the application, was
information requested by Italy (EC-96/At.9) necessary to ensure that conclusions of the safety
assessment were valid?
Dr. Nutti
758. Based on Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of
Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003), paragraphs 53, my understanding
is that additional animal feeding studies may be warranted for GM foods if changes in the
biovailability of the nutrients are expected or if the composition of the GM food is not comparable to
conventional food. So, the request from Italy for more animal feeding studies was not necessary to
ensure that conclusions of the safety assessment were valid. My understanding is that the dossier
presented followed not only the OECD consensus document but also the Codex guidelines, as far as
nutrition, composition and toxicology is concerned. It is clear for me that the substantial equivalence
studies compared the GM with the conventional counte rpart. Therefore, my opinion is that the Italian
request on these topics was not necessary to ensure that conclusions on the safety assessment were
valid. I can not answer on the questions related to molecular characterization (a to h).
Bejo-Zaden Transgenic Radicchio rosso
(EC chronology 97)
Question 55: Given the information before the Panel, including the application (EC-97/At.1),
and additional information provided by the notifier (EC-97/At.25-28), was additional
information requested by the SCF (EC-97/At.20, 22-24, 28, 31, and 30), regarding molecular
characterisation, antibiotic resistant marker genes, and substantial equivalence necessary to
ensure that conclusions of the safety assessment were valid?
Dr. Nutti
759. Based on the information presented in EC-97/At.1, 25 and 28, I will comment on the
substantial equivalence item requested by SCF. The notifier has conducted several field tests from
1994 to 1996, where different characteristics were observed (colour, size of the plants, productivity
etc). The notifier presented some data on the com position for substantial equivalence in the first
dossier. In EC97, At. 28, the SCF informs that th e previous information was formally accepted and
requested analysis on crude protein and amino acid composition, carotenoids, folic acid and
vitamin C. My opinion is that this request is correct, since these parameters are important for the
nutritional evaluation of the product.

Bejo-Zaden Transgenic Green hearted chicory
(EC chronology 98)
Question 56: Given the information before the Panel, including the application (EC-98/At.9)
and additional information provided by the notifier (EC-98/At.35-37 and 39), was additional
information requested by the SCF (EC-98/At.30, 32-33, 34, 38, 40 and 41), regarding molecular
characterisation, antibiotic resistant marker genes, and substantial equivalence necessary to
ensure that conclusions of the safety assessment were valid?
Dr. Nutti
760. Based on the information presented in EC-98/At.9, 35 and 37, I will comment on the
substantial equivalence item requested by the SCF. The notifier has conducted several field tests from
1994 to 1996, where different characteristics were observed (colour, size of the plants, productivity
etc). The notifier presented some data on the composition for the substantial equivalence assessment
in the first dossier. In EC97, At. 33, the SCF stat es that substantial equivalence was claimed on the
basis of a very small sample under conditions that have not been clearly described and asks the
applicant to confirm substantial equivalence over two growing seasons and in six geographical
locations. The SCF also requested care to ensure that appropriate comparators were used and that
these were compared with an appropriate sample size at each location. The SCF stated that if
substantial equivalence should not be established, further detailed safety examination of the crop must
be undertaken. I agree with the SCF comments and request in At 33.
761. In At 35, the applicant informs that the in formation on substantial equivalence was given
before, as information on chemical composition, taste tests, phenotypical description of breeding lines
and varieties, seeds and so on and informs that the data produced on crude protein, amino acid
composition, biogenic amines, carotenoids, vita mins and bitter compounds provided in may 1999
should be seen as additional to the information given before. The applicant pointed out that all this
information indicates that the GM chicory is equivalent to the conventional counterpart and there is no
sense in continuing with experiments for longer periods.
762. In At 38, the SCF informs that the previ ous information submitted by the notifier on
November 2000 was formally accepted and requested analysis on crude protein and amino acid
composition, carotenoids, folic acid and vitamin C. As I could not find this information on the files
submitted by the notifier, my opinion is that this request is correct, as this parameters are important
for the nutritional evaluation of the product.
763. My conclusion is that the request from SCF re garding substantial equi valence was necessary
to ensure that conclusions of the safety assessment were valid.

Pioneer LibertyLink and Bt (T25 x MON 810) corn (stack)
(EC chronology 101)
Question 57: Given the information before the Pa nel, including the application (EC-101/At.1-3)
and the additional information provided by Pioneer (EC-101/At13), was additional information
molecular characterization, field trials, seconda ry plant metabolites, and toxicological tests
requested by the Netherlands (EC-101/At.14) necessa ry to ensure that conclusions of the safety
assessment were valid?
Dr. Nutti
764. Based on the information presented in EC-96/At.1-3, At 13, I will comment on the
compositional analysis and toxicological analysis requested by The Netherlands (EC-101/At.14):
765. Question 2 is about field trials and compar ison between the composition of untreated and
ammonium glufosinate treated samples. In my opinion, the required information is not relevant to
ensure that conclusions of the safety assessment were valid, as in At-13, as I agree with the notifiers'
explanation that the aim of the safety assessment was not the ammonium glufosinate, but the new
product, obtained by a crossing T25xMON 810 corn.
766. Question 3, requests information on the levels of ferulic acid, p-coumaric acid, raffinose and
inositol, as the Committee believes that determining the levels of these plant metabolites is important
to rule out fortuitous changes in the plant meta bolism as result of the genetic modification. My
opinion is that this information required is not re levant to ensure that conclusions of the safety
assessment were valid, as the compositional informati on supplied before leads to the conclusion that
the product was substantial equivalent. I am also of the opinion that in this case, we have a product
that is a conventional cross of two GM corns, but this product has not been submitted to a new genetic
modification, so the possibility of fortuitous changes in the plant seems to be vanishing small, if
exists.
767. Question 4, on toxicological tests, asks for a semi -chronic oral toxicity study on mice or rats
with maize grain or meal from the hybrid lines, in order to provide additional data of the food safety
of this hybrid. I would like to recall the Code x Alimentarius Guidelines for the Conduct of Food
Safety Assessment of Foods Derived from Recomb inant-DNA Plants (CAC/GL 45-2003), paragraphs
53, that states that additional animal feeding studi es may be warranted for GM foods if changes in the
bioavailability of the nutrients are expected or if the composition of the GM food is not comparable to
conventional food. Therefore, the request from the Netherlands for semi-chronic oral toxicity study on
mice or rats with maize grain, were not necessary to ensure that conclusions of the safety assessment
were valid, as the data provided by the notifiers confirmed that T25 and MON 810 are substantially
equivalent to other commercial maize. In partic ular, it was shown the absence of expression of any
other sequence or fragment of the sequence inserted in T25 and MON 810. A far as the new proteins
expressed in T25 and MON 810 are concerned, for toxicological assessment were presented studies
on protein digestion in mammalian gastric and int estinal systems, acute gavage studies in mice,
homology to known toxins and alle rgens and exposure to human diet, that I understand are sufficient
for the toxicological assessment. In addition, a poultry feeding study has been carried out with grain
from T25xMON810, T25, MON 810 and non GM control with comparable genetics. The results
presented showed that no significant differences were observed on mortality, body weight grain and
feed conversion between chickens fed a diet containing maize grain from T25xMON810, T25,
MON 810, the non GM control or a standard diet cont aining yellow dent maize. I am of the opinion
that this additional study confirms that th e conventionally derived T25xMON 810 maize is
nutritionally equivalent to T25, MON810 and maize from non-GM co mmercial hybrids, so it would

not be necessary to carry out a semi-chronic oral toxi city study with maize or meal, as this study with
broilers has already provided the information with the grain.
Dr. Andow
Necessity to ensure validity:
768. In this question, I will address field trials, secondary plant metabolites, and toxicological tests.
769. The questions from the lead CA are reproduced here (EC-101/At.14, 23 April 2001).
2. The field trials. The Committee would appreciate receiving the results of the
comparison between the composition of untreated and glufosinate-ammonium treated
T25 maize by Aventis, as mentioned in your letter.
3. Secondary plant metabolites. In our previous letter we requested information
on the levels of ferulic acid, p-coumaric acid, raffinose and inositol. The Committee
believes that determining the levels of these plant metabolites is important to rule out
fortuitous changes in the plant metabolism as a result of the genetic modification.
The Committee has not received this information on either the hybrid or both parental
lines.
4. Toxicological tests. Although your remarks about the repeated backcrossing
to the non-GM inbred line have been noted, the Committee is still convinced that a
semi-chronic oral toxicity study on mice or rats with maize grain or meal from the
hybrid line is required to provide additional proof of the food safety of this hybrid.
770. The necessity of question 2 on composition of treated and untreated T25 maize is difficult to
determine. The comparison between untreated T25 maize and untreated maize lacking T25 would
seem to provide the necessar y evidence that T25 does or does not affect the composition of maize.
However, as indicated by the lead CA, it is also pos sible that the application of the herbicide could
affect the composition of T25 maize. The notifier makes a compelling point that the herbicide
registration process should account for all possible a dverse effects of the herbicide. However, the
herbicide registration process does not require demons tration of substantial equivalence. Moreover,
the herbicide is normally used with the T25 maize. Thus, there is a scientific justification for
requiring comparison of T25 with glufosinate, T25 without glufosinate and maize without T25 and
without glufosinate. However, I believe that there is a legitimate scientific debate as to whether all
three are necessary to ensure that conclusions of the safety assessment were valid.
771. The necessity of information about secondary plant metabolites can be justified by the
following argument. There is a legitimate concern that there are additional transgenes incorporated
into the T25 and Mon810 lines that could be ex pressing additional but unknown gene products, and
that elude detection by present molecular methods. In addition, it is also possible that the main gene
products from T25 or Mon810 could interact with pl ant metabolism, changing the composition of the
plant. The likelihood of the possibilities may be small, but it is difficult to argue how small.
Secondary plant compounds include many chemicals that are human health hazards at high enough
concentrations. The requested information can be considered to guard against possible changes in
human health hazard, and therefore can be cons idered necessary to ensure conclusions of the
safety assessment were valid. However, it is also possible that requests for information change in a
way that is not entirely justified. In this case, this does not appear to be the problem.

772. My response to this question is identical to my response to question 44 in paragraph 700.
This is not necessary to ensure conclusions of the safety assessment were valid.
773. The notifier submitted the original notification the the NL-CA on 20 April 2000. On 17 July
2000 the NL-CA requested additional information. On 22 November 2000 the notifier submitted
responses to this request. On 23 April 2001 the NL-C A asked for additional information. From the
time of original submission to the first request fo r additional information, 88 days passed. From the
time of submitting responses to the second request fo r additional information, 152 days elapsed,
including the winter holidays. This is a total of 240 days for review. This seems to be too long, even
taking into account delays associated with the winter holidays.
Dr. Squire
774. Similar difficulties are evident in the exchanges to those in 37-39 and elsewhere! Even so, the
requests by the Netherlands are arguably consistent with the type of information indicated in the
Codex Guideline for the conduct of food safety assessment of fo ods derived from recombinant-DNA
plants.
Monsanto/Syngenta Roundup Ready sugar beet (77)
(EC chronology 102)
Question 58: Given the information before the Panel, including the application
(EC-102/At.1-20) and the information provide d by Monsanto/Novartis (EC-102/At.22, 26 and
27-30), was additional information regarding food safety assessment of derived proteins
requested by the Netherlands (EC-102/At.32) necessa ry to ensure that conclusions of the safety
assessment were valid? Did the food safety a ssessments provided by the applicant follow the
Codex Guideline for the Conduct of Food Safety Assessment of Foods Derived from
Recombinant-DNA Plants.
Dr. Nutti
775. Based on the information submitted by application EC-102/At.1-20 and further information
provided by Monsanto/Novartis (EC-102/At.22, 26 and 27-30), my understanding is that the
information requested by the lead CA in EC- 102/At .32, regarding to the derived proteins and the
request for a semi-chronic oral toxicity test on mice or rats with edible parts of sugar beet, was not
necessary to ensure that the conc lusions of the safety assessment were valid. The applicants had
performed 90-day acute toxicity test with rats.
776. Based on the Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of
Foods Derived from Recombinant-DNA Plants (CAC /GL 45-2003), paragraphs 34 to 43, Assessment
of Possible Toxicity, we have at para 37 that the use of appropriate conventional toxicology or other
studies on the new substance may be necessary if, taking into account its function and exposure,
doubts about the safety of the new substance remain. In this case, the appli cant has provided in the
dossier (EC-102/At.1-20) all the information usua lly requested for the food safety assessment. Also,
for the toxicological assessment he presented safety studies on protein digestion in mammalian gastric
and intestinal systems, acute gavage studies in mice, homology to known toxins and allergens, and
exposure to human diet.
777. Based on the Codex Alimentarius Guidelines for the Conduct of Food Safety Assessment of
Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003), paragraphs 53, my understanding
is that additional animal feeding studies may be warranted for GM foods if changes in the

biovailability of the nutrients are expected or if the composition of the GM food is not comparable to
conventional food, that is not the case of Monsanto/Syngenta Roundup Ready sugar beet.
778. It was also confirmed that the GM crop was equivalent in composition and nutrition to the
conventional counterpart. Therefore, in my opinion, there was no need for requesting a semi chronic
toxicity study in mice or rats using the edible pa rts of the sugar beet in order to rule out possible
undesired effects of additional unidentified changes.
ISSUE 2
Background: In cases where a biotech product has obtained approval for Community-wide
marketing, individual EC member States may under certain circumstances provisionally prohibit the
marketing of that product in their own territori es. Such measures are commonly referred to as
"safeguard" measures.

It would be useful for the Panel to understand,

(a) For each biotech product for which a safeguar d measure was taken by specified EC member
States, how the scientific or other docume ntation relied upon by these member States
compares with:

 (i) the International Standard for Phytosanitary Measures (ISPM) No. 11 for Pest Risk
Analysis for Quarantine Pests, including its Supplement (EC-130);

 (ii) FAO/WHO Codex Principles for the Ri sk Analysis of Foods Derived from Modern
Biotechnology (attached);

 (iii) FAO/WHO Codex Guideline for the Conduct of Food Safety Assessment of Foods
Derived from Recombinant-DNA Plants (attached);

 (iv) FAO/WHO Codex Annex on Possible Allergenicity Assessment (attached); and

 (v) Annex III of the Cartagena Protocol on Biosafety to the Convention on Biological
Diversity (EC-1).

(b) For each product for which a safeguard me asure was taken by one of the specified EC
member States, whether the scientific or other documentation relied upon by these member States is
sufficient to support the safeguard measures taken.

The SPS Agreement defines risk assessment as:

"The evaluation of the likelihood of entry, establishment or spread of a pest or
disease within the territory of an importi ng Member according to the sanitary or
phytosanitary measures which might be applied, and of the associated potential
biological and economic consequences; or the evaluation of the potential for adverse
effects on human or animal health aris ing from the presence of additives,
contaminants, toxins or disease-causing or ganisms in food, beverages or feedstuffs."
(Annex A, paragraph 4)

Furthermore, the SPS Agreement provides that:

"In the assessment of risks, Members sha ll take into account available scientific
evidence; relevant processes and production methods; relevant inspection, sampling
and testing methods; prevalence of specific di seases or pests; existence of pest- or
disease-free areas; relevant ecologica l and environmental conditions; and
quarantine or other treatment." (Article 5.2)
and that:

"In assessing the risk to animal or plant life or health and determining the measure to
be applied for achieving the appropriate level of sanitary or phytosanitary protection
from such risk, Members sha ll take into account as relevant economic factors: the
potential damage in terms of loss of production or sales in the event of the entry,
establishment or spread of a pest or disease; the costs of control or eradication in
the territory of the importing Member; and the relative cost-effectiveness of
alternative approaches to limiting risks." (Article 5.3)
Questions
Oilseed rape MS1 x RF1 (notification C/UK/94/M1/1)
Safeguard measure of France
Question 59: Given the information before the Pa nel, including the evaluations undertaken by
the UK (EC-161/At.1), the Scientific Committee on Plants in May 1998 and May 1999
(CDA-35-A and CDA-69, respectively), and the European Commission in its Decision of
February 1996 (CDA-62), as well as th e information submitted by France
 with respect to its
safeguard measure (EC -161/At.3-11; CDA-68, CD A-70, CDA-71), is there any reason to believe
that the scientific evidence available to France in November 1998 and July 2001 was NOT
sufficient to permit it to undertake an appropriate assessment of potential risks to human, plant
and animal health, and the environment from th e growing for seed production of oilseed rape
MS1 x RF1? If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in November 1998 and/or in July 2001, was
there sufficient evidence available to France in August 2003
 to permit it to
undertake a more objective assessment of potential risks to human, plant and
animal health, and the environment from the growing for seed production of
oilseed rape MS1 x RF1? If not, what scientific evidence do you believe was
insufficient?
Dr. Nutti
779. EC161 At. 07–BEC considers that the knowledge which has been acquired makes it possible
to contemplate cultivating genetically modified, herbicide-tolerant beet, subject to code of practice
being applied and biovigilance measures being institu ted. It requires large scale experiments with
GM herbicide tolerant oilseed rape, in order to validate methods of managing crops of these varieties.
Finally, BEC recommends that overall consideration be given to dealing in detail with the question of
introducing new weed control practices which are b ased on the large scale use of total herbicides.
EC161–At. 07 – Summary of BEC, states that it is possible to contemplate cultivating genetically

modified, herbicide-tolerant beet, subject to code of practice being applied and biovigilance measures
being instituted.
EC161–At. 08 – Detailed opinion of BEC
780. Based on the information before the Panel, in cluding the evaluations undertaken by the UK
(EC-161/At.1), the Scientific Committee on Plan ts in May 1998 and May 1999 (CDA-35-A and
CDA-69, respectively), and the European Commi ssion in its Decision of February 1996 (CDA-62)
submitted by application (EC-102/At.1-20) and furt her information provided by Monsanto/Novartis
(EC-102/At.22, 26 and 27-30), my understanding is that there was enough scientific evidence
available to France in November 1998 and July 2001, information sufficient to permit it to undertake
an appropriate assessment of potential risks to human , plant and animal health, from the growing for
seed production of oilseed rape MS1 x RF1. Environm ental issues are not my field, so I cannot judge
this point, but all the information for the safety assessment of potential risks to human and animal
health that was submitted by the applicants and evaluated by UK and SCP were adequate and in
accordance to the Codex Guidelines (FAO/WHO Codex Principles for the Risk Analysis of Foods
Derived from Modern Biotechnology; FAO/WHO Code x Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible
Allergenicity Assessment).
781. It is important to point out that either the l ead CA (UK) and the SCP concluded that there is
no evidence to indicate that the placing on the mark et of the oilseed rape MS1 x RF1, with the
purpose to be used as any other oilseed rape is likely to cause adverse effects on human health and on
the environment.
Dr. Andow
782. France provided reasons for its safeguard measur e, beginning in 1998 with EC-161/At.2trans.
Here France indicates that the spread of herbicide resistance and the concomitant loss of efficacy of
herbicides with small environmental impact are their concerns at the time of implementing the
safeguard measure. Specifically France says (in translation)
• Controlling adventitious flora a nd oilseed rape regrowth in plots in the event of crop
rotation and in peripheral areas, would be mo re complex. Consequently, systematic
recourse to molecules with less favourable ecotoxicological profiles may be a
necessity;
• the use of molecules with a favourable ecot oxicological profile may be lost because
of their proven loss of effectiveness, whereas these molecules are es pecially used for
weeding between crops, at field borders and in various environments such as edges of
railway tracks or roads.
The possible developments described above would be worsened by:
• The possibility of an accumulation of non- selective herbicide tolerance genes in
double-resistant single lines;
• systematic introduction of these genes in all cultivated species.
783. France also adds that "There is also the ques tion of the impact on agriculture of a possible
dependency vis-à-vis only two types of genes and active ingredients."

784. At the center of this is that France believes th at the probability of escape of the HT transgene
as oilseed rape volunteers or into wild species is not accurately assessed, and that the efficacy and
implementation of risk management measures to eliminate HT volunteers and wild species are not
accurately evaluated. On the probability of escape, France is concerned because the scientific
evidence available at the time comes from a few small-scale dispersal experiments. France believes
that when GMHT oilseed rape is commercialized, it will occur at a large spatial scale. Dispersal
processes at large spatial scales may exhibit different characteristics than those at small spatial scales.
This is contrary to the Opinion of the SCP,
114 although the SCP does not provide a scientific argument
against France's concern about spatial scale. France is also concerned that specifying and
implementing the good agricultural practices that would eliminate HT volunteers and wild species
will be considerably more difficult than merely allowing present agricultural practices to continue in
the expectation that present practices will eliminate these undesired plants. This directly contradicts
the SCP's opinion,
115 which seems to imply that normal agricultural practices will be sufficient.
785. In the study of dispersal in ecology, durin g the 1980s and 1990s it became increasingly
apparent that measures of dispersal rates and disp ersal kernels (dispersal probability as a function of
distance) were related to the spatial scale of the measurement. Estimated dispersal rates are smaller
when the spatial scale of measurement is smaller. This is probably a general result of the fact the
most biological dispersal kernels are highly lept okurtic. This scale-dependence has also been
observed for pollen dispersal from oilseed rape. One study from 1995 suggested this 116, and was later
confirmed in a 1999 117 and 2002 publication in Science 118 on large-scale pollen dispersal. Thus, the
scientific evidence in 1998 on di spersal of oilseed rape pollen w as not sufficient to complete an
accurate assessment of dispersal probability. Sufficient data appeared by the end of 2002.
786. The scientific basis for believing that normal "good" agricultural practices would be sufficient
to eliminate HT volunteers and weedy relatives was not clearly articulated prior to France's 1998
decision. It is likely that the scientific re asoning for believing that normal "good" agricultural
practices would be sufficient and easy to implem ent was as follows. Presently (meaning in 1998)

114 CDA-69 SCP Opinion, 18 May 1999 on France's invocation of Article 16 on MS1 x RF1.
115 CDA-69 SCP Opinion, 18 May 1999 on France's invocation of Article 16 on MS1 x RF1. The SCP
states "The available evidence from the scale of release at that time [19 May 1998] suggested that volunteers can
be controlled by agronomic practice (cultivation and the use of an alternative broad spectrum herbicide)
provided that adequate m onitoring procedures are in place to iden tify spillage, dispersal and any subsequent
volunteers." The reference to 19 May 1998 is to CDA-35A SCP Opinion, 19 May 1998 This Opinion is on
MS8 x RF3, not MS1 x RF1. The SCP considers that resistance can be managed by a cohesive code of practice
supported by a strong education program. Specifically they recommend
"2. The Committee was also of the opinion that the potential transfer of the herbicide resistance gene to
wild Brassica relatives is a new issue in Europe in view of the limited scale of release to date. The Committee
has examined the available evidence from monitoring and research programmes to date. After evaluating all the
information available to the Committee, it was concluded that herbicide-tolerant volunteers that may appear
would be canola plants and not wild Brassica relatives. Such herbicide-tolerant volunteers could be controlled
in subsequent crops by conventional agricultural methods. The Committee recommends that the introduction of
herbicide-tolerant crops should be accompanied by:
i) an agreed code of practice for the particular m odified crop involving the active participation of the
notifier to promote best practice by farmers.
ii) a monitoring programme with an agreed design and implementation plan to detect the occurrence
and the establishment of herbicide-tolerant volunteers and weeds under field conditions in the EU."
116 Timmons et al. 1995 Euphytica 85: 417-423.
117 Thompson et al. 1999 in EC-162/At.13.
118 Pollen-Mediated Movement of Herbicide Resistan ce Between Commercial Canola Fields Mary A.
Rieger, Michael Lamond, Christoph er Preston, Stephen B. Powles, and Richard T. Roush Science 28 June
2002; 296: 2386-2388.

oilseed rape volunteers are normally not significant we eds, and in the rare case when they become
weeds, they can be easily controlled using th e proper herbicide. Weedy Brassica species are
important weeds, but these can also be controlle d using appropriate agricultural methods. Accepting
that this argument is true for the purposes of this paragraph, what remains uncertain is whether the
level of weed control that is considered sufficien t for normal agricultural practice is sufficient for
eliminating HT volunteers and wild species to the level desired by France.
787. Alternatively, the argument might not be true. There may be years when certain fields of
GMHT oilseed rape are not harvested at all and the entire crop goes to seed (since this happens
occasionally for non-transgenic oilseed rape). Th e subsequent Brassica weed problem may require
exceptional agricultural practices to bring the fi eld under control. Such possibilities were not
considered by the SCP, and France may be concerned about such scenarios.
788. Finally, France may be convinced (rightly, I think) that agricultural management of HT
volunteers and wild species will not be as simple as the SCP believes. Hence the scientific evidence
in 1998 on the agricultural practices necessary to eliminate HT volunteers and wild species was not
sufficient to complete an accurate assessment of risk management practices. Based on the material in
EC-161, I could not determine when sufficient inform ation was first available. In 2001 the French
Biomolecular Engineering Committee (BEC) called for additional research in this area, but by the
middle of 2003, the BEC believed that it had sufficient information to complete an assessment.119
789. France also could have argued in 1998 that ther e was insufficient data to identify all of the
possible environmental hazards of GMHT oilseed rape. France could have argued that the molecular
characterization was insufficient to identify all transgene products. It is not clear to me if MS1 x RF1
hybrids were ever subject to the molecular an alyses performed for the GM crops evaluated under
Directive 2001/18/EC. France could have argued that the risk of evolution of resistance in weeds not
related to the Brassicaceae had been incompletely assessed and management of these risks required
additional scientific data. These concerns were wi dely appreciated in the early 1990s, but action to
address them in the EU for GMHT crops did not take place until sometime after 1998. It is not clear
to me that adequate information to address this issu e exists even as of this date in 2005. France could
have argued that the data on non-target effects was insufficient. This claim would have been borne
out first by the suggestion in 2000 that GMHT oilseed rape could adversely affect skylarks in the UK
followed by the FSE trials in 2003 that suggested some possible adverse effects. Sufficient data were
available to assess these risks by October, 2003. Finally, recently France appears to argue that
contamination of conventional o ilseed rape varieties is an important concern (related to the
"coexistence" issue). While this risk seems possibl e to assess now, it is not clear that the scientific
information for justifying the risk management practices is available (this is because I am uncertain
about the details of the information available about good agricultural practices noted in
paragraph
788.

119 EC-161/At.3trans.

Dr. Snow
BACKGROUND FOR MY ANSWER

To explain my answer to this question, I include relevant statements from France, the European
Commission, and the SCP opinions, along with a few comments below.

1. France

France stated (EC-161/At.3 translation): "These orders banning the placing on the market of
genetically modified oilseed rape are based on the risk of contamination of crops of conventional
oilseed rape by transgenic oilseed rape. In an opinion given in this regard on 16 February 2001, the
Commission du Génie Biomoléculaire (Biomol ecular Engineering Committ ee) concluded that
experimentation should be pursued in order to supplement existing scientific knowledge and validate
methods for managing the cultivation of genetically modified oilseed rape."

Also, " A further referral to the Bi omolecular Engineering Committee by the French authorities on
26 June 2003 aimed to take stock of existing knowle dge on the gene flow in oilseed rape and of the
identification of risks connected with large-scale cultivation of genetically modified herbicide-tolerant
oilseed rape varieties."

In addition, in EC-161/At. 4 translation (see excerp ts under Question 60), France listed environmental
concerns and specific scientific investigations that were needed to address these concerns.

2. Early European Commission Decision on gluf osinate-tolerant oilseed rape (Feb. 6, 1996)

This favorable decision to place seeds of MS1BnxRF1Bn on the market states that:

"the survival and spread of this herbicide-to lerant swede-rape was evaluated, as well
as the transfer of the herbicide tolerance gene or the other modified genes to
compatible species; whereas it was concluded that the risk of establishment was low
and that any spread or transfer of the herbicide tolerance gene could be controlled by
using existing management strategies."
As I discuss elsewhere, the assumption that " the risk of establishment is low" is not supported by
scientific evidence, including evidence that became available during 1998-2003.

3. Opinions of the Scientific Committee on Plants

These opinions are nearly identical to others deali ng with glufosinate-tolerant oilseed rape. They
consider that gene transfer and persistence in wild Brassica rapa is unlikely, that potential problems
with volunteers can be managed, and that any problem s with the spread of herbicide-resistant weeds
are agronomic problems rather than environmental on es. The SCP requested that a code of practice
and monitoring plans be established to determine wh ether problems emerge with herbicide-tolerant
volunteers and wild relatives.

Opinion of the Scientific Committee on Plants regarding the glufosinate to lerant, hybrid rape derived
from genetically modified parental lines (MS8 x RF3) notified by Plant Genetic Systems (notification
C/B/96/01) (Submitted by the Scientific Committee on Plants, 19 May 1998

Excerpts – "The risk assessment assumes that tr ansfer will occur at a low level. The
relevant question is whether this can be contained by risk management and whether it
is an environmental or agronomic problem."
"Available evidence from the scale of release to date suggests that volunteers can be
controlled by agronomic prac tice (cultivation and the use of an alternative broad
spectrum herbicide) provided that adequat e monitoring procedures are in place to
identify spillage, dispersal and any subsequent volunteers. Normal management
methods for wild Brassicae including cultiv ation, rotation and alternative herbicide
should be maintained."
"Modified rape is no more invasive than unm odified plants and can be controlled by
the combination of cultivation and the use of alternative non-selective herbicides.
Potential transgenic exchange is unlikely to lead to establishment as a result of
reduced viability of any hybrid plants and competition."
Opinion of the Scientific Committee on Plants , adopted on 18 May 1999, on the Invocation by France
of Article 16 ('safeguard' clause) of Council Directive 90/220/EEC with respect to a genetically
modified oilseed rape notification C/UK/94/M1/1 (Plant Genetic Systems N.V.)

Excerpts – "In its risk assessment the Committee assumed that any such transfer will
occur at a low level. It therefore considered the more relevant question, whether this
can be contained by risk management and whether it is an environmental or
agronomic problem."
"The information provided by the French Competent Authority was taken into
consideration by the Committee in formulating its original Opinion in 1998 and is not
considered to change the environmental assessment and advice to the Commission."
Answer to Question 59:

790. Given that,
 (i) the SCP identified concerns about the spread of herbicide resistance to volunteer
weeds, even though they considered this to be more of an agronomic problem than an
"environmental" one;

 (ii) the SCP underestimated the extent of gene flow that is to be expected to other oilseed
rape crops, volunteer weeds, and weedy Brassica rapa; and

 (iii) the SCP recommended that the introduction of herbicide-tolerant crops should be
accompanied by "ii) a monitoring programme with an agreed design and
implementation plan to detect the occurrence and the establishment of herbicide-
tolerant volunteers and weeds under field conditions in the EU;"

791. I conclude that France had valid reasons to fo llow the advice of its Biomolecular Engineering
Committee to carry out more research to "suppl ement existing scientific knowledge and validate
methods for managing the cultivation of genetically modi fied oilseed rape." Scientists in France were
well aware of the fact that transgenes would be dispersed by means of pollen and seeds, making it
difficult to design management plans that would prevent problems in the future. These problems
could be compounded if transgenes for glyphosate tolerance were also approved in oilseed rape and

other crops, as I explain elsewhere. One could argue that future events are not relevant to this isolated
product, but glyphosate-tolerant oilseed rape has also been proposed for commercialization in Europe.
792. SUB-QUESTION TO 59. – If so, what scientific evidence do you believe was insufficient?
793. My conclusion is that France needed more info rmation about the rate at which the transgene
for glufosinate tolerance would spread to volunteer plants (including the seed bank) and related
weeds. France wanted this information in order to determine whether the types of problems that
Canada was experiencing (see above) could be managed in the context of the France's agricultural
system and, if so, what the cost would be to farmers.
794. By 2003, more scientific knowledge was availabl e in the form of peer-reviewed papers and
reports of farmers' experiences in Canada, but ne w information was still coming to light about the
spread and persistence of genes for herbicide tolerance (see Context section above, and References).
Dr. Squire
795. The view expressed by France was that insufficient information was available to provide clear
conclusions on several aspects of cross-pollination and persistence of GMHT oilseed rape. The EC
Scientific Committee on Plants had considered that the risks arising from outcrossing to volunteer,
feral and wild relatives were defi nite, but small because any progeny could be controlled later in the
rotation. France appeared to take a stronger line, in dicating the risks were larger or more uncertain.
The EC SPC and the information submitted by France therefore agreed on the nature of this potential
problem, but differed on its importance or extent . In November 1998 and July 2001, there was
knowledge about the mechanisms invol ved in cross-pollination and persistence, but one of France's
main arguments was based on the up-scaling effect – the uncertainty of what might occur if GMHT
crops were grown widely in the country (Notes, paragraphs 11, 12). There was insufficient knowledge
at that time to be able to predic t accurately, for a country such as France, what the rates of spread and
cross pollination would be (GM to non-GM) if a large part of the rapeseed areas were GM.
796. (a) The documentation is also correct when it states that relevant new experimental
information was being placed before the public (see example references below). The new information
was providing greater knowledge of cross-pollination in oilseed rape, particularly over distances of
several kilometres. This new information would no t have provided qualitativ ely different knowledge
(it was already known that oilseed rape could cr oss-pollinate at distance) but would provide better
quantitative estimates of the pollination frequencies and the subsequent survival of GM hybrids
among volunteer populations. Even by the end of 2003, there was still uncertainty as to what the field-
to-field cross pollination might be if GMHT crops came to be grown in many fields throughout a
landscape in Europe. Data in Europe was confined to knowledge of crossing from a relatively small
source (one or a few fields of donor oilseed rape) to a large potential sink (many fields on recipient
oilseed rape). Work is still in progress, not least as one of the main aims of the SIGMEA project
(Notes, paragraph 14), to estimate spread and cross-pollination on a regional scale in Europe.
120

120 Rieger MA, Lamond M, Preston C, Powles SB, Roush R. 2002. Pollen-mediated movement of
herbicide resistance between commercial canola fields. Science 296, 2386-2388.
Ramsay G, Thompson CE, Squire GR. 2003. Quantifying landscape-scale geneflow in oilseed rape.
Final report on Project RG 0216, 48 pp. Defra: London.

Question 60: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by France evaluate the relevant risks to human, plant or animal health, and the
environment from the growing of oilseed rape MS1 x RF1 for seed production?
(a) How does the scientific evidence and oth er documentation submitted by France
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
797. The reasons given by France were mainly relate d to the code of practice being applied and
biovigilance measures being instituted. These are environmental issues that I cannot answer but,
again, I would like to point out that there was no scientific evidence regarding to food and feed safety
assessment, for animal and human consumption, on which France could be based for asking the
moratorium.
Dr. Andow
798. With reference to the SPS Agreement, France has evaluated relevant risks to plant health, and
expressed some of these risks in terms of their consequent potential effect on human health.
Specifically, France has taken into account available scientific evidence (Article 5.2).
799. It is not clear, however, that France conduc ted the risk assessment according to the
phytosanitary measures which might be applied. France did not present their safeguard measure as a
possible phytosanitary measure, and did not conduct the risk assessme nt according to their safeguard
measure. Thus it is not clear that France conduc ted the risk assessment consistent with Annex A,
paragraph 4.
800. France did take into account relevant economic factors (Article 5.3), however France did not
explicitly compare the relative cost-effectiveness of alternative approaches to limiting risks.
801. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case because
France argues that there is a plant pest risk.
802. For those risks within the scope of ISPM 11 the risk assessment process is consistent with
ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
803. It appears that much of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of France contradict this guidance. My reading of the
materials before the Panel is that France did not explicitly address this guidance. Specifically I note
Section 3, S1 (measures should be designed in proportion to the risk), Section 3.1 (level of acceptable
risk should be expressed), Section 3.4 (principle s), and Section 3.4.6 (on prohibition) were not
explicitly addressed.
804. The science, documentation and reasoning of Fr ance are consistent with Annex III of the
Cartagena Protocol on Biosafety to the Convention on Biological Diversity. Specifically, Annex III,
Section 8(f) states "Where there is uncertainty re garding the level of risk, it may be addressed by
requesting further information on the specific issues of concern or by implem enting appropriate risk
management strategies and/or monitoring the living modified organism in the receiving environment."

Dr. Snow
805. With reference to the SPS definition of a risk assessment, in EC-161/ At. 4, France stated:
"The main undesirable effects expected from the ma rketing of genetically modified herbicide tolerant
oilseed rapes are therefore agronomic and commercial."
806. To the extent that these concerns are justified in the context of invoking "safeguard" measures
(which I consider to be a legal question), I conclu de that France had valid reasons for deciding that
additional scientific research was needed. My reasons are explained below.
807. Briefly, France was concerned about the spread of herbicide-tolerant volunteers and related
weeds, such as Brassica rapa. This concern was mentioned repeatedly by the EC Scientific
Committee on Plants, although the SCP felt that the ri sk could be managed after commercialization of
the product. The French Biomolecular Engineering Committee felt that this should be studied before
rather than after de-regulation. This is understa ndable if one assumes that the spread of herbicide
tolerance genes is harmful to the environment and to farmers (see above), because after the crop has
been released it could be very difficult to er adicate unwanted transgenes from volunteer, weedy, or
cultivated populations.
808. Based on excerpts from the documentation from France and the USA explanation of risk
assessments, both of which are included below, the requested research can be viewed as part of a
standard risk assessment. The requested research focuses on the "exposure" term reflecting how
widely a given harm will occur (as in risk = exposure x harm).
809. The First Written Submission of the USA, point 103, explains that risk assessments should
"(2) evaluate the likelihood of entry, establishment or spread of these diseases [or pests], as well as
the associated potential biological and economic consequences." Detailed empirical studies about the
spread of herbicide-tolerance genes by pollen and inter-specific hybrids are useful for estimating 1)
how common these genes could become in volunteer and weed populations, 2) whether this problem
can be mitigated, and 3) what type of monitoring should be considered after commercial release has
occurred.
810. In EC-161/At. 4 translation, France stated:
"Oilseed rape is widely grown in France, covering 1.1 million hectares in 2003,
nearly 3 per cent more than in 2002. It r anks after wheat (4.6 million hectares) and
maize (1.8 million hectares). ..."
"The main undesirable effects expected from the marketing of genetically modified
herbicide tolerant oilseed rapes are therefore agronomic and commercial. It would
appear that dispersal, at a rate to be spec ified, of the herbicide tolerance genes into
the environment towards adventitious flora is likely. Furthermore, dispersal into the
environment through introspecific hybridiz ation is inevitable, particularly on the
borders of fields and along lanes and roads. Oilseed rape is a highly competitive
species and has the capacity to generate regrowth for several yea rs in the following
rotation crops ..."
"Accordingly, in the context of a referral from the Ministry of Agriculture on 6 July
2000, the Biomolecular Engineering Committee issued an opinion on 16 February
2001 (attached), in which it concluded that experimentation should be pursued so as
to supplement existing scientific knowle dge and validate methods for managing the

cultivation of genetically modified oilseed rape. It found in particular that additional
knowledge was needed on:
– The effects that would be produced by sources of pollen larger in size than
those which had hitherto generated information on the dispersal of oilseed
rape pollen;
– pollen dispersal over a long distance, and particularly the role of insect
transfer;
– better characterization of particular in terspecific hybrids (amphidiploids), in
the interests of a better understanding of the species' dynamics;
– the importance of the impact of islands of persistent oilseed rape on the
pollination of neighbouring oilseed rape fields, including on those that would
result from the escape of seeds in the transport phases, and the dynamics of
such islands (persistence, extension, ... )."
"A further referral by the Ministry of Ag riculture to the Biomolecular Engineering
Committee, on 26 June 2003, aimed to ta ke stock of acquired knowledge regarding
gene flow in oilseed rape and of the identification of risks relat ed to large scale
cultivation of genetically modified herbicide tolerant oilseed rape varieties."
811. Regarding Question 60 (a), I did not find an I SPM statement about situations that would
trigger "safeguard measures" while additional scientific studies are carried out.
Dr. Squire
812. As indicated earlier, the requirements in the international guidelines are quite stringent. It
could be argued that France's position was compatible with the tone of the SPS Agreement, Annex A,
paragraph 4 (including economic as well as biological consequences) and compatible also with
ISPM-11 Annex 3 on 'Determining the potential for a LMO to be a pest'.
Question 61: Does the scientific evidence and ot her information submitted by France support
the adoption of a temporary prohibition on the growing of oilseed rape MS1 x RF1 for seed
production? In light of any potential risks identified by France, what other risk management
options were available in November 1998 and/or July 2001? What other risk management
options are now available?
Dr. Nutti
813. The reasons given by France were mainly relate d to the code of practice being applied and
biovigilance measures being instituted. These are environmental issues that I cannot answer, but again
I would like to point out that there were no scientific evidence regarding to food and feed safety
assessment, for animal and human consumption, on which France could be based for asking the
moratorium.
Dr. Andow
814. The adoption of a temporary prohibition can be justified on the basis of the scientific evidence
and other information submitted by France.

815. Several other risk management options could also have been justified in November 1998 and
July 2001. Risk management strategies include risk avoidance, risk mitigation, and risk tolerance. In
1998 and 2001 mitigation and toleranc e strategies were probably inappr opriate. Here I consider only
the risk that France expressed in its original decision to impose a temporary prohibition. One risk
avoidance strategy would have been to allow limited planting in a restricted region. In the first year,
this could have been at a scale of a large field trial, and build up from th ere. This would allow
determination of scale effects. Another strategy is outlined in paragraph 78.
816. Today several alternative risk management options are available. In addition to risk
avoidance as in the previous paragraph, risk mitigat ion strategies may also be possible to control HT
volunteers and wild species. It would appear that France is now convinced that if HT volunteers or
wild species occur that they can be detected rapidly enough and eliminated.
Dr. Snow
817. I do not have enough information to answer this question. If the question refers to seed
production on a very small scale, a temporary pr ohibition presumably would be less urgent and
possibly unnecessary. If the plants are grown on a fairly large scale, for example on 1,000 hectares
per year, the issues at hand are similar to those for the previous questions.
818. The basic concern is that herbicide resistance genes would spread in conjunction with wider
use of this herbicide over time, and that other ty pes of herbicide resistance genes might also be
approved in the future (e.g., glyphosate resistan ce), thereby compounding the problem. One could
argue that future events are not relevant to this isolated product, but glyphosate-tolerant oilseed rape
has also been proposed for commercialization in Europe.
Dr. Squire
819. The risk management options were and are similar to those indicated generally for oilseed
rape at Responses 6d and 101. The risk of spread and cross-pollination can be reduced by such
measures but not eliminated.
Topas 19/2 (notification C/UK/95/M5/1)
Question 62: Given that oilseed rape Topas 19/2 was only approved for import of seeds for
processing, what is the relevance of the findings of the UK farm scale evaluation study (EC-38)
in terms of the assessment of potential risks from Topas 19/2?
Dr. Nutti
820. In the study presented in EC-38, the researchers monitored plants and animals in the field and
around ploughed edges of the fields called field ma rgins. This was done before, during and after the
crops were grown and harvested. It was an environmental study, in order to give information for the
environmental assessment. In my opinion, this study has no relationship with the safety assessment of
Topas 19/2, as the permission for this product was asked for "import of seeds for processing", and not
for cultivation in Europe. My conclusion is that this study has no releva nce for the assessment of
potential risks of a product that will be only processed and not cultivated in Europe.

Dr. Andow
821. The findings of the UK FSE study (EC-38) are not relevant to the risk assessment for import
and processing of Topas 19/2. Their only possible re levance would be in the hazard identification
step of risk assessment. However, it is unclear how Topas 19/2 could reach such widespread levels of
actual (accidental) cultivation to cause such a h azard. It would require escape during importation
and/or processing and several years of multiplication at levels similar to the multiplication during
oilseed rape seed production and the growing of th ese large quantities on the landscape. Such large
quantities would necessitate actual cultivation, presu mably without the knowledge that the seed is
derived from Topas 19/2. Even if such a scenario were possible, there would many, many possible
ways to manage this risk.
Dr. Snow
822. Because the seeds of Topas 19/2 were not inte nded for cultivation, the UK farm scale
evaluation study bears little or no relevance to this decision. I assume that any seed spillage that
might occur when the seeds are delivered could be mana ged, and that it is extremely unlikely that the
seeds would be cultivated inadvertently. (It appears that Topas 19/2 was approved for cultivation in
the UK in 1996 (EC-162 At. 002-003), but I assume that this earlier decision is not applicable here.)
823. In EC-162 At. 007, Greece states that "it is certain that the seeds will escape into the
environment and will give viable plants" in Greece. Even if this is the case, I do not think the UK
farm scale evaluations are relevant to the possib ility that feral populations of herbicide-tolerant
oilseed rape would become established. The farm scale evaluations would only be relevant if the
product were cultivated widely.
Dr. Squire
824. Topas 19/2 and the Farm scale evaluations (FSE).
825. The FSE compared the effects of GMHT and conventional weed management. Its results
should have no bearing on the risk associated w ith the importation of seeds for processing. Even if
these seeds were spilled and became part of the wa yside feral population or volunteer population in a
field, they would have no effect unless the management of the field changed away from current
management in a way comparable to the differe nces between GMHT and conventional cropping in
the FSE.
Safeguard measure of France
Question 63: Given the information before the Pa nel, including the evaluations undertaken by
the UK (EC-162/At.1 to 3), and the SCP in February 1998 (CDA-63), May 1999 (CDA-65 and
CDA-73), and the European Commission in its Decision of April 1998 (CDA-61), as well as the
information submitted by France
 with respect to its safeguard measure (EC-162/At/5, EC-
161/At.3-11, CDA-64, CDA-66, CDA-67), is there an y reason to believe that the scientific
evidence available to France in November 1998 and July 2001 was NOT sufficient to permit it to
undertake an appropriate assessment of potential risks to human, plant and animal health, and
the environment from the import and processing of oilseed rape Topas 19/2? If so, what
scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in November 1998 and/or in July 2001, was
there sufficient evidence available to France in August 2003
 to permit it to

undertake a more objective assessment of potential risks to human, plant and
animal health, and the environment from the import and processing of oilseed
rape Topas 19/2? If not, what scientific evidence do you believe was insufficient?
Dr. Nutti
826. Answer 63: In EC-162/At.003, the lead CA (UK), having assessed the potential risks to
human health and environment arising from the ge netic modification, proposes to consent to the
placing on the market of the product for specific uses , such as import of Gm seed for processing of
food, animal feed and industrial uses of non-li ving processed products arising from the product;
growing and multiplication of seed for breeding material and for sale; field cropping for grain
production for feed, food and industrial uses of non-living processed products arising from the
product and had specified the conditions and labelling for this uses.
827. In CDA-63, it can be observed that in February 1998 the SCP made it clear that the safety
evaluation was done for importation of seeds only, for placing on the market for food and feed
purposes. The SCP concludes that there was no significant risk to human and livestock following
ingestion of CM seeds. They considered that the GM product and products th ereof were substantially
equivalent to the traditional counterpart. SCP concluded that there was no evidence to indicate that
the placing on the market of the oilseed rape Topas 19/2, with the purpose to be imported for
processing and placing on the market for food and f eed uses, is likely to cause adverse effects on
human health and on the environment.
828. On CDA-65, it can be observed that on May 1999 the SCP was consulted by the Commission
on the dossier for Topas 19/2 and the prohibitio n from the French CA, with concerns on the
environmental impact of genetic escape through volunteers and hybrids, and the agricultural
consequence of the spread of herbicide tolerance in both cultivated and non-cropped habitats. In its
comments, the SCP emphasizes that the application was for the importation of oilseed rape into
Europe for processing and not for cultivation and production within member states. The SCP also
explains that the potential for the loss of material during transport and the possible establishment of
feral plants in uncultivated habitats was considered in the risk assessments carried out by SCP in
forming its opinion of February 1998. They concluded that the risk of genetic escape from this market
consent was considered by the SCP to be smal l and the current information submitted by French
authorities did not change that assessment.
829. Based on the information presented by the notif iers and by the evaluation of SCP, my
understanding is that there is NO reason to believe that the scientific evidence available to France in
November 1998 and July 2001 was NOT sufficient to permit it to undertake an appropriate
assessment of potential risks to human, plant and animal health, and the environment from the import
and processing of oilseed rape Topas 19/2.
830. Answer 63a: In my opinion, the scientific evidence presented was sufficient in 1998 and
2003.
Dr. Andow
831. This is a notification for importation a nd processing, not for cultivation (despite
EC-162/At.1-3 stating that the UK evaluates it for cultivation).
832. France relies on the same rationale here as us ed for MS1 x RF1 (Questions 59-61), which I
have reviewed previously in paragraphs 782-783 and will not repeat here. The key difference is that

MS1 x RF1 was for cultivation and seed production as well as importation and processing. There are
also minor, formal differences in the Opinion of the SCP that have no consequence for this
discussion.121
833. The argument used by France th at was based on spatial scale does not hold for this case.
With importation and processing, it is highly unlikely that large-scale production of GMHT oilseed
rape is possible. Thus, the data on small-scale di spersal is probably sufficient to complete a risk
assessment even in 1998.
834. Moreover, the hazard of widespread HT resistance in oilseed rape or related wild species does
not hold for this case. This hazard is also scale-dependent, and with the small-scale releases and poor
survival expected in spillage and other accidental releases, it is not feasible that this hazard will occur.
Even if it were possible (see paragraph
821), there would many, many other possible ways to manage
this risk short of a prohibition. Thus, the rationale stated by France does not argue that a risk exists
that requires management.
835. France also could have argued in 1998 that ther e was insufficient data to identify all of the
possible environmental hazards of GMHT oilseed rape. (1) France could have argued that the
molecular characterization was insufficient to identify all transgene products. France could have
concluded that there may be risks stemming from these unidentified transgene products (if they exist)
that would require a safeguard measure. It is not clear to me if Topas 19/2 was ever subject to the
molecular analyses performed for the GM crops evaluated under Directive 2001/18/EC. (2) Recently
France appears to argue that contamination of conventional oilseed rape varieties is an important
hazard (related to the "coexistence" issue). This is a hazard that could occur from the small-scale
releases expected by accidental spilla ge. The risk associated with this hazard is probably possible to
assess now, but I do not have access to the scientific information on the risk management practices
available to me to make a more certain statement.
836. Such possible hazards would have allowed France to argue that there is a risk, and that the
risk management measures proposed by the SC P were not supported sufficiently by science. 122 The
scientific evidence in 1998 on the agricultural practices necessary to eliminate HT volunteers and wild
species resulting from spillage may not have been su fficient to complete an accurate assessment of
risk management practices. Based on the materi al in EC-161 and EC-162, I could not determine
when sufficient information was first available. Th e information needed would be substantially less
than for MS1 x RF1.
837. Given the information before the Panel, Fran ce could have argued in November 1998 that
there was insufficient evidence to complete a risk assessment. However, I believe that had France
collected the necessary data to complete this ri sk assessment promptly after implementing their
temporary prohibition that they could have had sufficient data to make a decision in 2001.
Dr. Snow
838. In summary, I did not find any convincing ar guments for why further scientific research
would be needed to investigate environmental effects of a product that is only intended for import and
processing and not for cultivation.

121 CDA-63, CDA-65, CDA-73.
122 This is discussed in more detail in my response to question 59, paragraph 786.

839. See ANSWER 62 for explanation. Because the seeds of Topas 19/2 were not intended for
cultivation, the need for environmental assessments a ppears to be very low to non-existent. I assume
that any seed spillage that might o ccur when the seeds are delivered could be managed and that it is
extremely unlikely that the seeds would be cultivated inadvertently. The only conceivable risk that I
can imagine is that feral volunteer populations would cross-pollinate with weedy Brassica rapa, giving
this weed the trait of resistance to glufosinate, and the weeds would become more abundant because
farmers would repeatedly spray these populations with glufosinate (killing off other weeds and
favoring the proliferation of the herbicide-resistan t weeds). This scenario seems extremely unlikely
for the case of GM seeds that are imported for processing and not cultivation.
840. I do not understand why France states " These orders banning the placing on the market of
genetically modified oilseed rape are based on the risk of contamination of crops of conventional
oilseed rape by transgenic oilseed rape" with regard to Topas 19/2 (EC-161 At. 003 translation).
841. ANSWER 63 (a): My conclusion is that I did not find any convincing arguments for why
further scientific research would be needed to inv estigate environmental effects of a product that is
only intended for import and processing and not for cultivation. See ANSWERS 62 and 63.
Question 64: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by France evaluate the relevant risks to human, plant or animal health, and the
environment from the import and processing of oilseed rape Topas 19/2?
(a) How does the scientific evidence and oth er documentation submitted by France
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
842. Environmental issues are not my field, so I can not judge this point, but all the information for
the safety assessment of potential risks to human and animal health that was submitted by the
applicants and evaluated by UK and SCP were adequate and in accordance to the Codex Guidelines
(FAO/WHO Codex Principles for the Risk Analysis of Foods Derived from Modern Biotechnology;
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible Allergenicity Assessment).
Dr. Andow
843. With reference to the SPS Agreement, France did no t identify a relevant risk to plant health.
However, it would have been possible to identif y a relevant risk to plant health (paragraph 789).
Specifically, France did not fully take into account av ailable scientific evidence (Article 5.2). As a
consequence, the argument was undercut by a faulty analysis and France missed opportunities to
make a stronger scientific argument.
844. Under the assumption that France had identified a relevant risk to plant health, it is not clear
that France conducted the risk assessment according to the phytosanitary measures which might be
applied. France did not present the safeguard me asure as a possible phytosanitary measure. France
did not conduct the risk assessment according to this proposed safeguard measure. Thus it is not clear
that France conducted the risk assessment consistent with Annex A, paragraph 4.

845. France did take into account relevant economic factors (Article 5.3), however France did not
explicitly compare the relative cost-effectiveness of alternative approaches to limiting risks.
846. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case. While
France did not identify a potential pest risk, it w as possible to identify a potential pest risk under
which ISPM 11 could be considered. Thus, ISPM 11 could be invoked.
847. For those risks within the scope of ISPM 11 the risk assessment process would be consistent
with ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
848. It appears that much of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of France contradict this guidance. My reading of the
materials before the Panel is that France did not explicitly address this guidance. Specifically I note
Section 3, S1 (measures should be designed in proportion to the risk), Section 3.1 (level of acceptable
risk should be expressed), Section 3.4 (principle s), and Section 3.4.6 (on prohibition) were not
explicitly addressed.
849. The science, documentation and reasoning of Fr ance are consistent with Annex III of the
Cartagena Protocol on Biosafety to the Convention on Biological Diversity. Specifically, Annex III,
Section 8(f) states "Where there is uncertainty re garding the level of risk, it may be addressed by
requesting further information on the specific issues of concern or by implem enting appropriate risk
management strategies and/or monitoring the living modified organism in the receiving environment."
Dr. Snow
850. ANSWERS 64 and 64 (a): My conclusion is that I did not find any convincing arguments for
why further scientific research woul d be needed to investigate environmental effects of a product that
is only intended for import and processing and not fo r cultivation, so the question about international
guidelines seems irrelevant. See ANSWER 63.
Question 65: Does the scientific evidence and ot her information submitted by France support
the adoption of a temporary prohibition on th e import and processing of oilseed rape Topas
19/2? In light of any potential risks identified by France, what other risk management options
were available in November 1998 and/or July 2001? What other risk management options are
now available?
Dr. Nutti
851. As far as my knowledge goes, the scientific ev idence submitted by France does not support a
temporary prohibition on the import and processing of oilseed rape Topas 19/2.
Dr. Andow
852. The scientific evidence and other information submitted by France does not support the
adoption of a temporary prohibition on the import and processing of oilseed rape Topas 19/2.
However, scientific evidence and other information before the Panel could be used to support the
adoption of a temporary prohibition on the import and processing of oilseed rape Topas 19/2. Even
on these grounds, a temporary prohibition probably could not be justified past 2001.

853. Several other risk management options could have been justified in November 1998. Risk
management strategies include risk avoidance, risk mitigation, and risk tolerance. In 1998 mitigation
and tolerance strategies were proba bly inappropriate. Another risk avoidance strategy would have
been to request additional molecular characteriz ation data and allow limited importation with
attendant monitoring so that the practical aspects of controlling escapes from spillage could
experienced.
854. These options were available in 2001 and remain t oday. In addition risk mitigation strategies
may also be possible to control HT volunteers and wild species.
Dr. Snow
855. In summary, I did not find any convincing ar guments for why further scientific research
would be needed to investigate environmental effects of a product that is only intended for import and
processing and not for cultivation, so the question a bout risk management options seems irrelevant.
See ANSWER 63.
Safeguard measure of Greece
Question 66: Given the information before the Pa nel, including the evaluations undertaken by
the UK (EC-162/At.1 to 3), and the SCP in February 1998 (CDA-63), May 1999 (CDA-65 and
CDA-73), and the European Commission in its Decision of April 1998 (CDA-61), as well as the
information submitted by Greece
with respect to its safeguard measure (EC-162/At. 4, CDA-72),
is there any reason to believe that the scientifi c evidence available to Greece in September 1998
was NOT sufficient to permit it to undertake an appropriate assessment of potential risks to
human, plant and animal health, and the envi ronment from the import and processing of
oilseed rape Topas 19/2? If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in September 1998, was there sufficient
evidence available to Greece in August 2003 to permit it to undertake a more
objective assessment of potential risks to human, plant and animal health, and
the environment from the import and processing of oilseed rape Topas 19/2 (EC-
162/At.6 to 13)? If not, what scientific evidence do you believe was insufficient?
Dr. Nutti
856. Answer 66: On CDA-73, the SCP answers the points risen by the Greek CA, and does not
change their previous risk assessment. The SCP ex plains that the potential for the loss of material
during transport and the possible establishment of fera l plants in uncultivated habitats was considered
in the risk assessments carried out by SCP in forming its opinion of February 1998. The SCP points
out that in the absence of the use of glufosinate ammonium to apply selective pressure, modified rape
is not more invasive than unmodified rape and o ilseed rape is not grown commercially in Greece.
They concluded that the risk of genetic escape from this market consent was considered by the SCP to
be small and the current information submitted by Greek authorities does not change that assessment.
857. Based on the information presented by notifiers and by the evaluation of SCP, my
understanding is there is NO reason to believe th at the scientific evidence available to Greece on
September 1998 was NOT sufficient to permit it to undertake an appropriate assessment of potential
risks to human, plant and anim al health, and the environment from the import and processing of
oilseed rape Topas 19/2.

858. Answer 66a: In my opinion, the scientific evidence presented was sufficient in 1998 and
2003.
Dr. Andow
859. This should be considered a continuation of my responses to Questions 62-65. In EC-
162/At.7 Greece identifies three risks to human, plant or animal health for which there is insufficient
evidence to undertake an appropriate assessment. These are: 1. harm of wildlife and the
environment;123 2. human health risk from outcrossing to wild Brassica species (e.g., Brassica
cretica) in Greece that are used as food; 3. in creased difficulty in managing farmland because GMHT
oilseed rape overwinters better in Greece, should spread faster and is hard to eliminate once
established as a weed.
860. In paragraph 821, I suggestted that the first of these is not relevant to Topas 19/2, which is a
notification for import and processing. The SCP Opinion states that there is no evidence that the
known gene products of the Topas 19/2 tr ansformation are a human health risk,
124 and Greece does
not address this explicitly. Thus, there is little meri t in the second risk. There is some merit in the
third risk. On this point there is a clear differen ce in the Opinion of the SCP, which considers this
remote, and Greece, which does not. Oilseed rape survives better in Greece than in France, so while
this risk is probably very small in France, it is likel y to be larger in Greece. It is probably fair to
conclude that it is not certain how much higher this may be. However, if this risk were substantially
larger than believed by the SCP, there would many ot her possible ways to manage this risk short of a
prohibition.
861. Greece also could have argued in 1998 that ther e was insufficient data to identify all of the
possible environmental hazards of GMHT oilseed rape. (1) Greece could have argued that the
molecular characterization was insufficient to iden tify all transgene products. Greece could have
concluded that there may be risks stemming from these unidentified transgene products (if they exist)
that would require a safeguard measure. It is not clear to me if Topas 19/2 was ever subject to the
molecular analyses performed for the GM crops evaluated under Directive 2001/18/EC. (2) Recently
Greece appears to argue that contamination of conve ntional oilseed rape varieties is an important
hazard (related to the "coexistence" issue). This is a hazard that could occur from the small-scale
releases expected by accidental spilla ge. The risk associated with this hazard is probably possible to
assess now, but I do not have access to the scientific information on the risk management practices
available to me to make a more certain statement.
862. Risk 3 (paragraph 859) and such possible hazards as in paragraph 861 would allow Greece to
argue that there is a risk, and that the risk management measures proposed by the SCP were not
supported sufficiently by science. 125 The scientific evidence in 1998 on the agricultural practices
necessary to eliminate HT volunteers and wild sp ecies resulting from spillage may not have been
sufficient to complete an accurate assessment of ri sk management practices. In addition, Greece may
have believed that implementing "good" agricultur al practices would be more complicated than
anticipated by the SCP. Based on the material in EC-161 and EC-162, I could not determine when
sufficient information was first available. The information needed would be substantially less than for
MS1 x RF1 (questions 59-61).

123 This complaint foreshadows the results from the UK-FSE trials and models of skylark impact
124 CDA-63, CDA-73.
125 This is discussed in more detail in my response to question 59, paragraph 786.

863. Given some of the information provided by Greece and other the information before the
Panel, Greece could have argued in November 1998 that there was insufficient evidence to complete a
risk assessment. However, I believe that had Greece collected the necessary data to complete this risk
assessment promptly after implementing their te mporary prohibition that they could have had
sufficient data to make a decision in 2001.
Dr. Snow
864. See ANSWERS 63-65. I did not find any convincing arguments for why further scientific
research would be needed to investigate environmen tal effects of a product that is only intended for
import and processing and not for cultivation. The specific arguments provided by Greece seem
flawed. I do not understand why Greece assumes th at feral populations would become established or
why they would cause environmental problems. Even if such populations become established, there
is no scientific reason to expect that gene flow coul d harm the genetic diversity or abundance of wild
relatives of oilseed rape in Greece. The arguments that Greece made for additional research appear to
be based on the assumption that the product would be cultivated rather than imported only for
processing.
Question 67: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Greece evaluate the relevant risk s to human, plant or animal health, and the
environment from the import and processing of oilseed rape Topas 19/2?
(a) How does the scientific evidence and oth er documentation submitted by Greece
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
865. Environmental issues are not my field, so I cannot judge this point, but all the information for
the safety assessment of potential risks to human and animal health that was submitted by the
applicants and evaluated by UK and SCP was adeq uate and in accordance to the Codex Guidelines
(FAO/WHO Codex Principles for the Risk Analysis of Foods Derived from Modern Biotechnology;
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible Allergenicity Assessment).
Dr. Andow
866. With reference to the SPS Agreement, Greece has identified one relevant risk to plant
health.126 In addition, it is also possible to identify ot her relevant risks to plant health. Specifically,
in some places Greece did not fully take into account available scientific evidence, but in others it did
(Article 5.2). In addition Greece missed opportunities to make scientific arguments in its favor.
867. It is not clear that Greece conducted the risk assessment according to the phytosanitary
measures which might be applied. Greece did not present the safeguard measure as a possible
phytosanitary measure, and did not conduct the risk assessment according to this proposed safeguard
measure. Thus it is not clear that Greece conduc ted the risk assessment consistent with Annex A,
paragraph 4.

126 Risk 3, paragraph 859.

868. Greece did take into account relevant economic factors (Article 5.3), however Greece did not
explicitly compare the relative cost-effectiveness of alternative approaches to limiting risks.
869. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case, because
Greece did identify a potential risk. In addition, ot her potential risks relevant to ISPM 11 could be
identified.
870. For those risks within the scope of ISPM 11 the risk assessment process is consistent with
ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
871. It appears that much of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of Greece contradict this guidance. My reading of the
materials before the Panel is that Greece did not ex plicitly address this guidance. Specifically I note
Section 3, S1 (measures should be designed in proportion to the risk), Section 3.1 (level of acceptable
risk should be expressed), Section 3.4 (principle s), and Section 3.4.6 (on prohibition) were not
explicitly addressed.
872. The science, documentation and reasoning of Greece are consistent with Annex III of the
Cartagena Protocol on Biosafety to the Convention on Biological Diversity. Specifically, Annex III,
Section 8(f) states "Where there is uncertainty re garding the level of risk, it may be addressed by
requesting further information on the specific issues of concern or by implem enting appropriate risk
management strategies and/or monitoring the living modified organism in the receiving environment."
Dr. Snow
873. See ANSWER 66.
Question 68: Does the scientific evidence and ot her information submitted by Greece support
the adoption of a temporary prohibition on the import and processing of oilseed rape
Topas 19/2? In light of any potential risks iden tified by Greece, what other risk management
options were available in September 1998? What other risk management options are now
available?
Dr. Nutti
874. As far as my knowledge goes, the scientific ev idence submitted by Greece does not support a
temporary prohibition on the import and processing of oilseed rape Topas 19/2.
Dr. Andow
875. Some of the scientific evidence and other information submitted by Greece supports the
adoption of a temporary prohibition on the import and processing of oilseed rape Topas 19/2. In
addition, scientific evidence and other information before the Panel could be used to support the
adoption of a temporary prohibition on the import and processing of oilseed rape Topas 19/2. Even
on these grounds, the temporary prohibition probably could not be justified past 2001.
876. Several other risk management options could have been justified in November 1998. Risk
management strategies include risk avoidance, risk mitigation, and risk tolerance. In 1998 mitigation
and tolerance strategies were proba bly inappropriate. Another risk avoidance strategy would have
been to request additional molecular characteriz ation data and allow limited importation with

attendant monitoring so that the practical aspects of controlling escapes from spillage could
experienced.
877. These options were available in 2001 and remain t oday. In addition risk mitigation strategies
may also be possible to control HT volunteers and wild species.
Dr. Snow
878. See ANSWER 66.
Maize Bt-176 (notification C/F/11-03)
Safeguard measure of Austria
Question 69: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-158/at. 1 to 3); the Scientific Committee for Animal Nutrition in December 1996
(EC-158/At. 4 and 5); the Scientific Committee for Food in December 1996 (US-64) and in
March 1997 (US-58); the Scientific Commi ttee for Pesticides in December 1996
(EC-158/At.6_SCI) and in May 1997 (US-57); the Scientific Committee for Plants (SCP) in
September 2000 (US-66); and the European Co mmission in its Decision of January 1997
(ARG-37), as well as the information submitted by Austria
 with respect to its safeguard
measure (US-52, EC-158/At.11, 12 and 15; EC-144, EC-147), is there any reason to believe that
the scientific evidence available to Austria in Febr uary 1997 was NOT sufficient to permit it to
undertake an appropriate assessment of potential risks to human, plant and animal health, and
the environment from the importation and use of Maize Bt-176? If so, what scientific evidence
do you believe was insufficient?
(a) If the evidence was not sufficient in February 1997, was there sufficient evidence
available to Austria in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from the importation and use of Maize Bt-176 (EC-158/At.30 to
42)? If not, what scientific evidence do you believe was insufficient?
Dr. Nutti
879. Answer 69: In EC158-At.04, the Scientific Committee for Animal Nutrition, concludes that
the probability of the transfer of a functional bl a-gene construct from GM maize to bacteria is
virtually zero and if this virtually impossible ev ent would take place, it would have no clinical
significance. The conclusion also pointed out that th ere is no evidence of a risk of causing B-lactam
antibiotic resistance in animal bacteria with the use of the GM maize. The SCAN opinion was that
there is no evidence indicating that the use in anim al feeding of the GM maize will give rise to any
adverse effect on animal health.
880. In EC158-At. 05, the Scientific Committee for F ood concludes that the GM maize is, except
for the inserted traits, substantially equivalent to maize present on the market. Animal feeding studies
support its substantial equivalence to the parent plant. No nutritional concerns are associated with the
use of this transgenic line; and it is unlikely that the genetic changes introduce any new potential for
allergenicity. No human toxicological concerns arise regarding the inserted traits based upon the
toxicological and degradation data considered. The conclusion also stated that the possibility that the
product would add significantly to the already widesp read occurrence of ampici llin resistant bacteria
in animals and man is remote and the risk of bacterial transformation is extremely low. The
committee recognized the general question of using genes coding for antibiotic resistance for the

development of novel foods and propose to scrutinise future needs and applications of this marker
genes. In March 1997 (US-58), the Scientific Committee for Food stated that the Austrian
information does not provide new scientific evidence regarding the food use of Ciba-Geigy maize that
was not taken into account by the Committee at the time its opinion was delivered. Thus, the Austrian
information does not cause the SCF to consider that the GM maize constitutes a risk to human health.
881. In EC158-At. 06_SCI, the Scientific Committee for Pesticides pointed out the glufosinate
ammonium was not authorized for direct applicatio n into maize plants, and should not, therefore, be
used for post emergence on genetically modified maize without authorization, in accordance with
provisions of Directive 91/414/EEC. As far as the Bt event is concerned, the Scientific Committee for
Pesticides was of the opinion that the possible development of insect resistance to the Bt toxin arising
from the cultivation of maize plants containing Bt -toxin would not be an adverse effect on the
environment.
882. In EC158-At. 07, the Austrian CA informs the applicant that the marketing of Ciba Geigy
Maize in Austria has been pr ohibited by an ordinance, which entered in force on 14 th of February of
1997, and this action has been in accordance with Art. 16 of Directive 90/220/EEC. The lead CA
recognizes that expert from different scientific committees have extensively reviewed and discussed
problems, concluding the lower risk of the product and all the scientific comments and arguments are
valid and well taken, however, from the Austrian poi nt of view, especially new scientific results have
questioned the present scientific possibility of co nclusive evaluation of the mechanism of gene
transfer as well as the development of resistance to Bt toxin.
883. The European Commission, in its Decision of Ja nuary 1997, has given consent to the placing
on the market of the Ciba Geigy Maize and any progeny derived from crosses of this product with any
traditionally bred maize.
884. Based on the information presented by the notifie rs and by the evaluation of four different
scientific committees, my understanding is there is NO reason to believe that the scientific evidence
available to Austria in February 1997 was NOT suffi cient to permit it to undertake an appropriate
assessment of potential risks to human, plant an d animal health, and the environment from the
importation and use of Maize Bt-176.
885. Answer 69a: In my opinion, the scientific evidence presented was sufficient in 1997 and
2003.
Dr. Andow
886. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
887. The initial assessment of Event 176 by France pr ovided in EC-158/At.1- 3 (10 January 1995,
1 March 1995, 3 March 1995) does not contain th e details of the assessment by the French
Biomolecular Engineering Committee (BEC).
888. The Opinion of the Scientific Committee on Pesticides (EC-158/At.6_SCI, 9 December 1996)
on Event 176 did not assess the risk of resistance evolution in the target pest s because it considered
this risk to be an agricultural risk, not an environm ental risk. In either ev ent, the Committee felt that
resistance management should be fully considered. The Committee also did not assess non-target
effects.

889. The Opinion of the SCAN (EC-158/At.4, 13 December 1996) did not address any
environmental risk, including environmentally medi ated indirect effects on animal health. The
Opinion of the SCF (US-64, EC-158/At.5, 13 December 1996) did not address any environmental
risk, including environmentally mediated indirect effects on human health.
890. On 23 January 1997, the EC decided to allow the placing on the market of Event 176
(ARG-37).
891. On 13 February 1997, Austria decided to ban the commercialization of Event 176 in Austria
(US-52). Austria gave three reasons for this action. 1. A concern that the risks of the bla gene
(ampicillin resistance) are greater than that assessed by the SCAN and the SCF (paragraph 889).
2. That the Cry1Ab toxin in Event 176 could have ri sks to non-target organisms, including those in
the soil such as collembola. 3. There is an envi ronmental risk that the target insects will evolve
resistance to the Cry1Ab toxin, and resistance management measures should be required (they were
not required by the EC).
892. In the Further Opinion of the Scientific Committee on Pesticides (US-57, 12 May 1997), "The
Committee concluded that the reasons and information submitted by the Austrian Authorities did not
add new relevant evidence to that already cons idered by the Committee and that none of its
conclusions on the risk to the environment were affected by the Austrian arguments."
893. The reasoning in the Further Opinion of the Scientific Committee on Pesticides (US-57,
12 May 1997) is false and did not take into account the scientific information known at the time. The
conclusions it reached (paragraph 892) do not follow from the evidence available at the time.
894. (1) On page 2, the Committee stated that it "recognized the complexity of comparing the
exposure of a pest to genetically modified plants , where exposure may be prolonged and maintained,
and conventionally applied pesticides, with shor ter and repeated exposure. The potential for
development of resistance could be either accel erated or retarded." (a) In its 9 December 1996
Opinion, the Committee nowhere recognized the comple xity of comparing the exposure of a pest to
genetically modified plants, where exposure may be prolonged and maintain ed, and conventionally
applied pesticides, with shorter and repeated expos ure. It must be granted that this is a new
consideration for the SCP that was introduced by Aust ria. (b) While it is true that theoretically the
difference in exposure could result in faster or slower resistance evolution, the SCP is not discussing a
theoretical case. There was more than adequate data available in 1995 from the notifier's efficacy
trials (and efficacy trials on Event 176 conducted by public sector scientists) and a long history of
efficacy trials of Bt insecticide sprays on maize to demonstrate without question that Event 176 would
exert a selection pressure on corn borers many times stronger (I would estimate ~1000x) than a
typical Bt insecticide spray. The conclusion of the Committee either is not case-specific and therefore
inappropriate, or it is case-specific and therefore false.
895. (2) On page 2, the Committee stated "Based on available information, soil exposure from the
GMO maize plants will be less than the expo sure resulting from a single conventional spray
application including the run-off from the plants. Only trace amounts of toxin can be detected in the
roots of the maize and furthermore, normal agricultu ral practice would involve removal of the greater
part of the plant at harvest. The plant remains ar e often shredded and transformed into silage for use
as animal feed at a later stage." (a) In its 9 December 1996 Opinion, the Committee nowhere made the
assessment that based on available information, soil exposure from the GMO maize plants will be less
than the exposure resulting from a single conventio nal spray application including the run-off from
the plants. Nowhere did the Committee note that th ese issues were addressed satisfactorily in the
dossier submitted by the notifier. It must be granted that these considerations are new to the

Committee and that they were introduced by Austri a. (b) The conclusion that soil exposure from the
Event 176 maize plants will be less than the ex posure from a single Bt spray application was
debatable even in May 1997. The Committee did not appear to take into account that nearly all of the
Cry toxin applied in a Bt spray would be inactivated by sunlight in less than week after spraying. The
Committee did not appear to consider that most fa rmers incorporate maize residue into the soil after
harvest. This would incorporate several tons of biomass per hectare, putting Cry toxin out of the sun,
where it could persist for some time. The Committe e seemed to believe that the maize residue would
be ensiled. Silage requires green plant material, and most silage maize is cut while still green, long
before it would be harvested for grain. Thus it is not clear that very much residue would be removed
for silage. Finally risk assessments should not rely on one study for key conclusions (in this case the
Palm et al. study, cited as study 4 in the Opin ion). The SCP assessment should have acknowledged
that scientific information was scarce at that time, and it would have been better had the SCP
maintained a more agnostic position with respect to the risk assessment. Thus, I conclude that the
Committee did not consider all available scientif ic evidence and did not appropriately weigh
potentially conflicting information when coming to their conclusion.
896. (3) On page 2-3, the Committee states "The Committee stated in its previous report of
9 December 1996 that resistance management strate gies are needed during the years of use of any
pesticide, Bt sprays included. The Committee drew attention once again to the need for effective
resistance management, including monitoring on ag ronomic grounds, to prolong the effectiveness of
Bt toxin both in conventional sprays and in genetically modified maize. It also felt that the
submission of a satisfactory monitoring and r esistance management programme should be a
requirement for the authorization to use genetically modified maize seeds expressing Bt-toxin." (a) In
its 9 December 1996 Opinion, the Committee nowhere made the recommendation that the submission
of a satisfactory monitoring and resistance manage ment programme should be a requirement for the
authorization to use genetically modified mai ze seeds expressing Bt-toxin. The acknowledgement
that it should be a requirement in this 12 May 1997 Opinion is a major change in the conclusions and
recommendations of the Committee. It must be gr anted that this change was due to Austria's
insistence that resistance management measures ar e required for commercial authorization. (b) The
Committee does not address Austria's main point on this issue, which is that resistance risk should be
considered an environmental risk. The Committee also seems to be unaware of the position of the
US-EPA in 1996, which was that resistance risk is an environmental risk. 127 This is a critical issue
because it determined whether resistance risk c ould be considered under Directive 90/220/EEC.
Thus, the Committee did not engage on a critical reason submitted by Austria.
897. (4) It must be granted that the reasons and information submitted by the Austrian Authorities
added new relevant evidence that the Scientific Committee on Pesticides had not considered in their
previous deliberation, and that that a key conclu sion was affected by the Austrian arguments.
Moreover, the key conclusions by the Committee in this Opinion on non-target risks are overstated
and/or false.
898. I conclude that the scientific evidence ava ilable to Austria in February 1997 was NOT
sufficient to permit it to undertake an appropriate assessment of potential risks to plants and the
environment from Event 176. The evidence on risks to non-target organisms, including those in the
soil such as collembola was insufficient. Austria could have mustered sufficient evidence to argue
that there was an environmental risk that the target insects will evolve resistance to the Cry1Ab toxin,
and resistance management measures should be re quired. There was probably insufficient evidence

127 Matten, S.R., et al., The U.S. Environmental Protection Agency's role in pesticide resistance
management, in Molecular Genetics and Evolution of Pesticide Resistance , T.M. Brown, Ed., American
Chemical Society, Washington, D.C., 1996, pp. 243-253.

available to determine what measures should be required. This determination by Austria was in direct
contradiction to the EC decision. In this case, Austria probably did not then and does not now need to
claim that their safeguard measure is a precautionary one.
899. By 2003, the basis for risk assessment of Bt crops had changed because several significant
scientific points had come to light. (1) It became widely appreciated that the molecular basis of
transformation was more complex than originally th ought, and the implications of these findings for
risk assessment were articulated. Annex II of Directive 2001/18/EC is one consequence, but
additional regulatory changes have occurred since then. Indeed, knowledge in molecular biology
continues to accumulate at remarkably fast rates, an d I expect that there will be continued change in
regulation in the future. 128 (2) Non-target risk assessment shifted from assessing indicators of
environmental risk to assessing actual identified poten tial environmental risks. Presently this is done
on an ad hoc basis, as no systematic methodology h as gained widespread acceptance. (3) Gene flow
risk assessment has shifted from being based primarily on an assessment of the probability of gene
flow to being based on an assessment of both the probability of gene flow and the conditional hazard
probability. (4) Resistance risk is considered an environmental risk, and science-based resistance
management measures are required.
900. Thus, evidentiary standards for what constitut es an objective environmental risk assessment
had changed substantially from 1997 to 2003. This is particularly true in Europe and the United
States, and particularly true for the Bt crops.
901. (1) Thus, in 2003, there remains some uncertainty about non-target risks of Event 176, but it
could be argued that a risk assessment could be conducted using scientifically justified worst-case
assumptions. On the other hand, Austria could reas onably maintain that there is still insufficient
information to know which non-target species may be at risk, and therefore it is not possible to
conduct an objective risk assessment. The findings in 2003 by Székács and Darvas (EC-158/At.37) –
that two protected butterfly species in Hungary, Inac his io and Vanessa atalanta, might be exposed to
Bt corn pollen and suffer higher mortality – certainly suggests that not all of the non-target species at
risk to Event 176 have been identified in Europe. (2) As in 1997, Austria could muster sufficient
evidence in 2003 to argue that there is an environmental risk that the target insects will evolve
resistance to the Cry1Ab toxin in Event 176, and resistance management measures should be required.
Moreover, in 2003, Austria should have sufficient evidence to determine what kind of resistance
management measures to require. (3) Unlike in 1997 , in 2003 Austria could argue that the molecular
characterization of Event 176 is insufficient to conduct a risk assessment. (4) In 2004, Austria
suggested that an additional gene flow risk that needs assessment is contamination of conventional
production. It is possible that the scientific evidence to support risk management measures for this
risk is available to conduct an objective risk assessment today.
Question 70: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Austria evaluate the relevant risks to human, plant or animal health, and the
environment from the importation and use of Maize Bt-176?
(a) How does the scientific evidence and oth er documentation submitted by Austria
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?

128 Possibilities are suggested in Nap et al. 2004. Genomics for Biosafety in Plant Biotechnology . IOS
Press, NATO Science Series, Amsterdam.

Dr. Nutti
902. Answer 70: Environmental issues are not my field, so I cannot judge this point, but all the
information for the safety assessment of potential risks to human and animal health that was submitted
by the applicants and evaluated by the Scientif ic Committee for Animal Nutrition, the Scientific
Committee for Food, the Scientific Committee for Pe sticides and the Scientific Committee for Plants
(SCP) were adequate and in accordance to the Codex Guidelines (FAO/WHO Codex Principles for
the Risk Analysis of Foods Derived from Modern Biotechnology; FAO/WHO Codex Guideline for
the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants;
FAO/WHO Codex Annex on Possible Allergenicity Assessment).
Dr. Andow
903. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
904. With reference to the SPS Agreement, Austria ha s evaluated relevant risks to plant health.
Specifically, Austria has taken into account available scientific evidence (Article 5.2).
905. It is not clear, however, that Austria conducted the risk assessment according to the
phytosanitary measures which might be applied. Austria did not present their safeguard measure as a
possible phytosanitary measure, and did not conduct the risk assessme nt according to their safeguard
measure. Thus it is not clear that Austria conducte d the risk assessment consistent with Annex A,
paragraph 4.
906. Austria did take into account relevant economic factors (Article 5.3), however it did not
explicitly compare the relative cost-effectiveness of alternative approaches to limiting risks.
907. The "potential pest" concept of ISPM 11 must fi rst be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case because
Austria argues that there is a plant pest risk.
908. For those risks within the scope of ISPM 11 the risk assessment process is consistent with
ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
909. It appears that some of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of Austria contradict this guidance. My reading of the
materials before the Panel is that Austria did not explicitly address this guidance. Specifically I note
Section 3.4 (principles), and Section 3.4.6 (on prohibition) were not explicitly addressed.
910. In US-52, Austria made it clear that they are applying a different standard for acceptable risk
than reflected in the assessments of the SCAN, SCF, Scientific Committee on Pesticides in the EC
decision (ARG-37) itself. In so doing, Austria fulfilled ISPM 11, Section 3.1 (level of acceptable risk
should be expressed).
911. ISPM 11, Section 3.4.6 (prohibition) states "If no satisfactory measure to reduce risk to an
acceptable level can be found, the final option ma y be to prohibit importation of the relevant
commodities. This should be viewed as a measure of last resort and should be considered in light of
the anticipated efficacy, especially in instan ces where the incentives for illegal import may be
significant." With respect to resistance risk, in 1997 Austria had established the existence of an
environmental risk, and required the development of measures to manage this risk to acceptable

levels. As the EC was not willing to make such a requirement, Austria had no choice but to intervene
with its own safeguard measure. In 1997, how ever, there was probably insufficient scientific
evidence to establish what necessary me asures should be taken. Scientif ically credible suggestions at
that time focused on using refuges (maize that w as not Bt maize) ranging from 10-70% of the maize
grown by any farmer who chose to grow Bt maize. It is unlikely that Austria could have implemented
any of these suggestions in time for the 1997 growin g season, so I conclude that Austria had fulfilled
ISPM 11, Section 3.4.6, at least for the 1997 growing season.
912. The science, documentation and reasoning of Aust ria are consistent with Annex III of the
Cartagena Protocol on Biosafety to the Convention on Biological Diversity. Specifically, Annex III,
Section 8(f) states "Where there is uncertainty re garding the level of risk, it may be addressed by
requesting further information on the specific issues of concern or by implem enting appropriate risk
management strategies and/or monitoring the living modified organism in the receiving environment."
Question 71: Does the scientific evidence and other information submitted by Austria support
the adoption of a temporary prohibition on the importation and use of Maize Bt-176? In light
of any potential risks identified by Austria, what other risk management options were available
in February 1997? What other risk management options are now available?
Dr. Nutti
913. It is important to point out that the main issu es raised by Austria were related to the present
scientific possibility of conclusive evaluation of the mechanism of gene transfer as well as the
development of resistance to Bt toxin. As far as my knowledge goes, concerning food safety and
nutrition, the scientific evidence submitted by Austria does not support a temporary prohibition on the
import and use of Maize Bt-176.
914. In EC-144, Austrian Contribution for the WTO-Dispute-Expert-Meeting of the European
Commission 14 January 2004 in Brussels, I found th at Austria presents also issues related to
allergenicity and toxicological ri sk assessment, but I cannot agree with the points raised, as they are
not in accordance with the FAO/WHO Codex Guide line for the Conduct of Food Safety Assessment
of Foods Derived from Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible
Allergenicity Assessment.
Dr. Andow
915. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
916. The adoption of a temporary prohibition can be justified on the basis of the scientific evidence
and other information submitted by Austria.
917. Other risk management options could not have been justified in February 1997. However,
had Austria worked to develop its own acceptable resistance management measures, perhaps by 1999
other risk management options would have been possible.
918. Today several alternative risk management options are available. Risk management strategies
include risk avoidance, risk mitigation, and ri sk tolerance. Tolerance strategies are probably
inappropriate. One alternate risk avoidance st rategy would be to im plement country-specific
resistance management measures, to limit planting to a restricted region, and to conduct intensive
non-target experiments. This would allow progressive determination of non-target effects.

Safeguard measure of Germany
Question 72: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-158/at. 1 to 3); the Scientific Co mmittee for Animal Nutrition in December 1996
(EC-158/At. 4 and 5); the Scientific Committee for Food in December 1996 (US-64) and in
March 1997 (US-58); the Scientific Committee for Pesticides in December 1996
(EC-158/At.6_SCI) and in May 1997 (US-57); the SCP in September 2000 (US-66); and the
European Commission in its Decision of Janua ry 1997 (ARG-37); as well as the information
submitted by Germany
 with respect to its safeguard measure (US-65, EC-158/At.18-29,
EC-144), is there any reason to believe that the scientific evidence available to Germany in
March 2000 was NOT sufficient to permit it to undertake an appropriate assessment of
potential risks to human, plant and animal heal th, and the environment from the importation
and use of Maize Bt-176? If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in Ma rch 2000, was there sufficient evidence
available to Germany in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from the importation and use of Maize Bt-176? If not, what
scientific evidence do you believe was insufficient?
Dr. Nutti
919. Answer 72: In US-65 (31/03/2000), the information submitted by Germany with respect to its
safeguard measure, my understanding is that the CA permits to sell seeds limited to 12 tons per year.
The reasons stated by Germany were that harmful effects exist for non-target organisms, development
of resistance, Bt toxin in the soil and antibiotic r esistant gene, so all points were environment related
issues.
920. Environmental issues are not my field, so I cannot judge this point, but all the information for
the safety assessment of potential risks to human and animal health that was submitted by the
applicants and evaluated by the Scientific Comm ittee for Animal Nutrition, the Scientific Committee
for Food, the Scientific Committee for Pesticides and the Scientific Committee for Plants (SCP) were
adequate and in accordance to the Codex Guide lines (FAO/WHO Codex Principles for the Risk
Analysis of Foods Derived from Modern Biot echnology; FAO/WHO Codex Guideline for the
Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO
Codex Annex on Possible Allergenicity Assessment).
921. Answer 72a: In my opinion, the scientific evidence presented was sufficient in 2000 and
2003, but I can answer only regarding food safety, a nd Germany points were related to environmental
issues.
Dr. Andow
922. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
923. The initial assessment of Event 176 by France pr ovided in EC-158/At.1- 3 (10 January 1995,
1 March 1995, 3 March 1995) does not contain th e details of the assessment by the French
Biomolecular Engineering Committee (BEC).

924. The Opinion of the Scientific Committee on Pesticides (EC-158/At.6_SCI, 9 December 1996)
on Event 176 did not assess the risk of resistance evolution in the target pest s because it considered
this risk to be an agricultural risk, not an environm ental risk. In either ev ent, the Committee felt that
resistance management should be fully considered. The Committee also did not assess non-target
effects.
925. The Opinion of the SCAN (EC-158/At.4, 13 December 1996) did not address any
environmental risk, including environmentally medi ated indirect effects on animal health. The
Opinion of the SCF (US-64, EC-158/At.5, 13 December 1996) did not address any environmental
risk, including environmentally mediated indirect effects on human health.
926. On 23 January 1997, the EC decided to allow the placing on the market of Event 176
(ARG-37).
927. Luxembourg (US-63, 7 February 1997) and Aust ria (US-52, 13 February 1997) decided to
ban the commercialization of Event 176 for reasons covered in questions 69 and 75.
928. The reasoning in the Further Opinion of the Scientific Committee on Pesticides (US-57,
12 May 1997) is false and did not take into account the scientific information known at the time. This
is fully discussed in my response to question 69, paragraphs 892-897, and will not be repeated here.
929. In my response to question 69 and question 75, I have concluded that the scientific evidence
available to Luxembourg and Austria in February 1997 was NOT sufficient to permit them to
undertake an appropriate assessment of potential risks to plants and the environment from Event 176.
930. The Regulatory Committee under Directive 90/220/EC met three times to consider Draft
Commission Decisions to require that the temporary prohibitions by Luxembourg and Austria be
repealed. The first two meetings (EC-158/At.13, 10 November 1997 and EC-158/At.16, 22 January
1998) resulted in decisions to delay making a deci sion. The final meeting (EC-158/At/17, 29 April
1998) resulted in no decision (neither to repeal nor not to repeal).
931. On 11 November 1997, an Expert Working Group on Bt resistance management was
launched. However, I cannot find any of the proceedings of this Working Group in the materials
before the Panel.
932. On 31 March 2000 (US-65), Germany decided to prohibit the unrestricted commercial use of
Event 176 in Germany. Germany considered the sci entific information for risk assessment to be
inadequate in the following areas: effects on non- target organisms, development of resistance,
countermeasures against development of resistance, effects of Bt toxin in the soil, horizontal or
vertical gene transfer of antibiotic resistance gene, harm to humans from antibiotic resistance gene.
933. On 9 November 2000 (US-66), the SCP provided an Opinion on the German decision. The
SCP concluded that the scientific information pr ovided by the German Competent Authority does not
alter the original risk assessments on Event 176. The Opinion of the SCP is one possible scientific
opinion that can be reached from the information available at the time, but the SCP should have
acknowledged that the new information also allows several other scientifically valid opinions that
were not justifiable in 1996.
934. On page 3-4 of this Opinion, the SCP deliberates on non-target risks, focusing on three
organisms, green lacewing, monarch butterfly and bl ack swallowtail. For the lacewing and monarch,
the SCP considered the experiments difficult to inte rpret and extrapolate to the field. It did not

interpret the swallowtail case. Rather than provid e a point by point discussion of the SCP Opinion, I
will list a series of valid scientific perspectives and interpretations on th e lacewing and monarch
studies that illustrate some of the diversity of valid scientific opinion that is not reflected in the SCP
Opinion. (1) According to the tiered non-target risk assessment protocols (which are consistent with
ISPM 11 and Annex III of the Biosafety Protocol and widely used in the US and Europe), the purpose
of laboratory experiments is to expose the orga nisms to concentrations higher than would be
considered typical for the field. By doing so, one reduces the probability of false negative effects.
Experimental positives then should undergo additional evaluation. (2) Both lacewings and monarchs
were exposed to concentrations of Cry1Ab toxin that would be expected to be higher than typical for
the field. Swallowtails were not exposed to high concentrations. (3) Both lacewings and monarchs
were adversely affected by Cry1Ab toxin in these laboratory experiments. (4) Additional assessment
should have been conducted on lacewi ngs and monarchs to determine the relevance to the field. (5)
Cry1Ab is supposed to be a toxin specific to mo ths and butterflies, but lacewings are not closely
related to moths and butterflies. Hence the toxi city spectrum of Cry1Ab toxin is broader than
previously expected. (6) Several years later, it h as been suggested that Event 176 would have caused
significant risk to monarchs had it become a popularly used variety.129
935. On page 4 of the Opinion, the SCP delib erates on resistance risk and management,
considering two organisms, European corn borer an d Mediterranean corn borer. The SCP advised on
the establishment of non-Bt refuges adjacent to modifi ed crops but pointed out that, in view of the
slow introduction into Europe, crops would be surrounded by natural refuges for some time to come.
Rather than provide a point by point discussion of the SCP Opinion, I will list a series of valid
scientific perspectives and interpretations that illustrate the prevailing scientific opinion on resistance
risk and management in 2000 that is not reflected in the SCP Opinion. (1) The rate of market
penetration after initial introduction of Bt maize in th e US was the fastest of any crop variety or crop
protection technology in the history of US agricultu re. Prior to introduction, many predicted that
market penetration would be slow. They were wr ong. (2) Resistance evolves locally. Thus refuges
must be available wherever Bt-maize is locally u sed. Thus, refuges need to be required from the
beginning. (3) Resistance management is the respons ibility of each farmer who uses Bt maize. Thus,
each farmer should be required to implement refuges. Thus, refuges need to be required from the
beginning.
936. On page 5 of the Opinion, the SCP delib erates on toxin release to soil, focusing on
degradation processes. The SCP suggests that because protein turnover occurs routinely in soils and
degradation of Bt toxin would be e xpected to degrade at rates simila r to other proteins or DNA in the
soil, there is no evidence that Bt-toxins will persis t in soils and have adve rse effects on non-target
organisms. Rather than provide a point by point discussion of the SCP Opinion, I will list a series of
valid scientific perspectives and inte rpretations that illustrate some of the diversity of valid scientific
opinion that is not reflected in the SCP Opinion. (1) The actual rates and degradation processes for
large proteins in soils is poorly understood. (2) Bt toxin loading in maize fields during and after
harvest can be substantial. Thus, scale effects are pos sible. (3) Presently it is known that Bt toxin in
the soil can have an adverse effect on earthworms. Whether this translates into an actual risk is not
yet known.
937. A close reading (paragraphs 934-936) of the SCP Opinion (US-66, 9 November 2000)
suggests that it has not considered all scientific perspectives (and in some cases ignored prevailing
scientific opinion). Thus, the SCP Opinion does not invalidate the scientific opinion of Germany.

129 NRC 2002. Environmental effects of transgenic plants: The scope and adequacy of regulation.
National Academy Press, Washington, DC. 320pp.

938. I conclude that the scientific evidence available to Germany in March 2000 was NOT
sufficient to permit it to undertake an appropriate assessment of potential risks to plants and the
environment from Event 176. The evidence on risks to non-target organisms, including those in the
soil could be considered insufficient. Germany coul d have mustered sufficient evidence to argue that
there was an environmental risk that the target insects will evolve resistance to the Cry1Ab toxin, and
resistance management measures should be requir ed. There may have been sufficient evidence
available to determine what measures should be required, but there probably was insufficient evidence
to determine how to implement these measures effect ively. This determination by Germany was in
direct contradiction to the EC decision. In this case, Germany probably does not need to claim that
their safeguard measure is a precautionary one.
939. By 2003, the basis for risk assessment of Bt crops had changed because several significant
scientific points had come to light. (1) It became widely appreciated that the molecular basis of
transformation was more complex than originally th ought, and the implications of these findings for
risk assessment were articulated. Annex II of Directive 2001/18/EC is one consequence, but
additional regulatory changes have occurred since then. Indeed, knowledge in molecular biology
continues to accumulate at remarkably fast rates, an d I expect that there will be continued change in
regulation in the future. 130 (2) Non-target risk assessment shifted from assessing indicators of
environmental risk to assessing actual identified poten tial environmental risks. Presently this is done
on an ad hoc basis, as no systematic methodology h as gained widespread acceptance. (3) Gene flow
risk assessment has shifted from being based primarily on an assessment of the probability of gene
flow to being based on an assessment of both the probability of gene flow and the conditional hazard
probability. (4) Resistance risk is considered an environmental risk, and science-based resistance
management measures are required.
940. Thus, evidentiary standards for what constitut es an objective environmental risk assessment
had changed substantially from 1997 to 2003. This is particularly true in Europe and the United
States, and particularly true for the Bt crops.
941. (1) Thus, in 2003, there remains some uncertainty about non-target risks of Event 176, but it
could be argued that a risk assessment could be conducted using scientifically justified worst-case
assumptions. On the other hand, Germany could reas onably maintain that there is still insufficient
information to know which non-target species may be at risk, and therefore it is not possible to
conduct an objective risk assessment. The findings in 2003 by Székács and Darvas (EC-158/At.37) –
that two protected butterfly species in Hungary, Inac his io and Vanessa atalanta, might be exposed to
Bt corn pollen and suffer higher mortality – certainly suggests that not all of the non-target species at
risk to Event 176 have been identified in Europe. (2) As in 1997, Germany could muster sufficient
evidence in 2003 to argue that there is an environmental risk that the target insects will evolve
resistance to the Cry1Ab toxin in Event 176, and resistance management measures should be required.
Moreover, in 2003, Germany should have sufficien t evidence to determine wh at kind of resistance
management measures to require. (3) Unlike in 1997, in 2003 Germany could argue that the
molecular characterization of Event 176 is insufficien t to conduct a risk assessment. (4) It is also
possible for Germany to suggest that an additional gene flow risk that needs assessment is
contamination of conventional production. It is po ssible that the scientific evidence to support risk
management measures for this risk is presently available.

130 Possibilities are suggested in Nap et al. 2004. Genomics for Biosafety in Plant Biotechnology . IOS
Press, NATO Science Series, Amsterdam.

Question 73: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Germany evaluate the relevant risk s to human, plant or animal health, and the
environment from the importation and use of Maize Bt-176?
(a) How does the scientific evidence a nd other documentation submitted by
Germany compare with the relevant intern ational guidelines for risk assessment
and analysis identified above?
Dr. Nutti
942. Answer 73: Environmental issues are not my field, so I cannot judge this point, but all the
information for the safety assessment of potential risks to human and animal health that was submitted
by the applicants and evaluated by the Scientif ic Committee for Animal Nutrition, the Scientific
Committee for Food, the Scientific Committee for Pe sticides and the Scientific Committee for Plants
(SCP) was adequate and in accordance to the C odex Guidelines (FAO/WHO Codex Principles for the
Risk Analysis of Foods Derived from Modern Biotechnology; FAO/WHO Codex Guideline for the
Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO
Codex Annex on Possible Allergenicity Assessment).
Dr. Andow
943. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
944. With reference to the SPS Agreement, Germany h as evaluated relevant risks to plant health.
Specifically, Germany has taken into account available scientific evidence (Article 5.2).
945. It is not clear, however, that Germany cond ucted the risk assessment according to the
phytosanitary measures which might be applied. Germany did not present their safeguard measure as
a possible phytosanitary measure, and did not conduct the risk assessment according to their safeguard
measure. Thus it is not clear that Germany conducted the risk assessment consistent with Annex A,
paragraph 4.
946. Germany did not take into account relevant economic factors (Article 5.3).
947. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case because
Germany argues that there is a plant pest risk.
948. For those risks within the scope of ISPM 11 the risk assessment process is consistent with
ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
949. It appears that some of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of Germany contradict this guidance. My reading of the
materials before the Panel is that Germany did not explicitly address this guidance. Specifically I note
Section 3.4 (principles), and Section 3.4.6 (on prohibition) were not explicitly addressed.
950. In US-65, Germany made it clear in some points that they are applying a different standard
for acceptable risk than reflected in the assessments of the SCAN, SCF, Scientific Committee on

Pesticides in the EC decision (ARG-37) itself. In so doing, Germany fulfilled ISPM 11, Section 3.1
(level of acceptable risk should be expressed).
951. ISPM 11, Section 3.4.6 (prohibition) states "If no satisfactory measure to reduce risk to an
acceptable level can be found, the final option ma y be to prohibit importation of the relevant
commodities. This should be viewed as a measure of last resort and should be considered in light of
the anticipated efficacy, especially in instan ces where the incentives for illegal import may be
significant." With respect to resistance risk, in 1997 Austria had established the existence of an
environmental risk, and required the development of measures to manage this risk to acceptable
levels. Germany concurred in 2000. As the EC was not willing to make such a requirement,
Germany had no choice but to intervene with its own safeguard measure. In 2000, however, there
was probably sufficient scientific evidence to esta blish what necessary measures should be taken.
Scientifically credible suggestions at that time focused on using refuges (maize that was not Bt maize)
ranging from 20-30% of the maize grown by any farmer who chose to grow Bt maize. It is unlikely
that Germany could have implemented any of th ese suggestions in time for the 2000 growing season,
so I conclude that Germany had fulfilled ISPM 11, Section 3.4.6, at least for the 2000 growing season.
952. The science, documentation and reasoning of Germ any are consistent with Annex III of the
Cartagena Protocol on Biosafety to the Convention on Biological Diversity. Specifically, Annex III,
Section 8(f) states "Where there is uncertainty re garding the level of risk, it may be addressed by
requesting further information on the specific issues of concern or by implem enting appropriate risk
management strategies and/or monitoring the living modified organism in the receiving environment."
Question 74: Does the scientific evidence and other information submitted by Germany support
the adoption of a temporary prohibition on the importation and use of Maize Bt-176? In light
of any potential risks identified by Germany, what other risk management options were
available in March 2000? What other risk management options are now available?
Dr. Nutti
953. It is important to point out that the main issues raised by Germany were that harmful effects
exist for non-target organisms, development of resist ance, Bt toxin in the so il and antibiotic resistant
gene, so all points were environmental related issu es. As far as my knowledge goes, regarding food
safety and nutrition issues, the scientific evidence submitted by Germany does not support a
temporary prohibition on the import and use of Maize Bt-176.
Dr. Andow
954. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
955. The adoption of a temporary prohibition (with < 500ha for research purposes) can be justified
on the basis of the scientific evidence and other information submitted by Germany.
956. Other risk management options could not have been justified in March 2000. However, had
Germany worked to develop its own acceptable r esistance management measures, perhaps by 2002
other risk management options would have been possible.
957. Today several alternative risk management options may be available. Risk management
strategies include risk avoidance, risk mitigation, and risk tolerance. Tolerance strategies are
probably inappropriate. One alternate risk avoidance strategy would be to implement country-specific

resistance management measures, to limit planting to a restricted region, and to conduct intensive
non-target experiments. This would allow progressive determination of non-target effects.
Safeguard measure of Luxembourg
Question 75: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-158/at. 1 to 3); the Scientific Co mmittee for Animal Nutrition in December 1996
(EC-158/At.4 and 5); the Scientific Committee for Food in December 1996 (US-64) and in
March 1997 (US-58); the Scientific Committee for Pesticides in December 1996
(EC-158/At.6_SCI) and in May 1997 (US-57); the SCP in September 2000 (US-66); and the
European Commission in its Decision of Janua ry 1997 (ARG-37), as well as the information
submitted by Luxembourg
 with respect to its safeguard measure (US-63, EC-158/At.9, EC-144),
is there any reason to believe that the scientifi c evidence available to Luxembourg in February
1997 was NOT sufficient to permit it to undertake an appropriate assessment of potential risks
to human, plant and animal health, and the environment from the importation and use of Maize
Bt-176? If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in February 1997, was there sufficient evidence
available to Luxembourg in August 2003 to permit it to undertake a more
objective assessment of potential risks to human, plant and animal health, and
the environment from the importation and use of Maize Bt-176? If not, what
scientific evidence do you believe was insufficient?
Dr. Nutti
958. Before judging the information submitted by Luxem bourg, it is important to point out that all
the information for the safety assessment of potential risks to human and animal health that was
submitted by the applicants and evaluated by the Scientific Committee for Animal Nutrition, the
Scientific Committee for Food, the Scientific Committee for Pesticides and the Scientific Committee
for Plants (SCP) were adequate, and the scientific opinion of these committees were that the product
could be planted and consumed in Europe.
959. In US-63, Ministerial Decree of 7 February 1997 prohibiting the use and sale of a corn
product obtained by genetic transformation, Minist er of Health, Luxembourg, considering that the
GM product contains the gene conferring resistance to ampicillin and that this circumstance risks
having implication for human health, that in the present state of the dossier cannot be properly
evaluated and considering that no monitoring prog ram has been implemented with regard to the
development of a resistance of the pyralid to the Bt-endotoxin, prohibits the use and sale of the Maize
Bt-176.
960. Based on the information presented by the notifie rs and by the evaluation of four different
scientific committees, my understanding is there is NO reason to believe that the scientific evidence
available to Luxembourg in February 1997 was NOT sufficient to permit it to undertake an
appropriate assessment of potential risks to human, plant and animal health, and the environment from
the importation and use of Maize Bt-176.
961. Answer 75a: In my opinion, the scientific evidence presented was sufficient in 1997 and
2003.

Dr. Andow
962. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks, specifically the resistance monitoring program.
963. The initial assessment of Event 176 by France pr ovided in EC-158/At.1- 3 (10 January 1995,
1 March 1995, 3 March 1995) does not contain th e details of the assessment by the French
Biomolecular Engineering Committee (BEC).
964. The Opinion of the Scientific Committee on Pesticides (EC-158/At.6_SCI, 9 December 1996)
on Event 176 did not assess the risk of resistance evolution in the target pest s because it considered
this risk to be an agricultural risk, not an environm ental risk. In either ev ent, the Committee felt that
resistance management should be fully considered.
965. The Opinion of the SCAN (EC-158/At.4, 13 December 1996) did not address any
environmental risk, including environmentally medi ated indirect effects on animal health. The
Opinion of the SCF (US-64, EC-158/At.5, 13 December 1996) did not address any environmental
risk, including environmentally mediated indirect effects on human health.
966. On 23 January 1997, the EC decided to allow the placing on the market of Event 176
(ARG-37).
967. On 7 February 1997, Luxembourg decided to ban the commercialization of Event 176 in
Luxembourg (US-63). Luxembourg gave two reasons for this action. 1. A concern that the risks of
the bla gene (ampicillin resistance) have not b een properly evaluated by the SCAN and the SCF
(paragraph 965). 2. That a monitoring program must be implemented to monitor the development of
resistance in the target pest to Cry1Ab toxin in Event 176.
968. The Further Opinion of the Scientific Comm ittee on Pesticides (US-57, 12 May 1997) is a
major shift in the position of the Committee on Event 176.
969. On page 2-3 of US-57, the Committee states "The Committee stated in its previous report of
the 9 December 1996 that resistance management st rategies are needed during the years of use of any
pesticide, Bt sprays included. The Committee drew attention once again to the need for effective
resistance management, including monitoring on ag ronomic grounds, to prolong the effectiveness of
Bt toxin both in conventional sprays and in genetically modified maize. It also felt that the
submission of a satisfactory monitoring and r esistance management programme should be a
requirement for the authorization to use genetically modified maize seeds expressing Bt-toxin." (a) In
its 9 December 1996 Opinion, the Committee nowhere made the recommendation that the submission
of a satisfactory monitoring and resistance manage ment programme should be a requirement for the
authorization to use genetically modified mai ze seeds expressing Bt-toxin. The acknowledgement
that it should be a requirement in this 12 May 1997 Opinion is a major change in the conclusions and
recommendations of the Committee. (b) The Committ ee does not address a significant issue implicit
in Luxembourg's statement, which is that resistance risk should be considered an environmental risk.
The Committee also seems to be unaware of the position of the US-EPA in 1996, which is that

resistance risk is an environmental risk. 131 This is a critical issue because it determined whether
resistance risk could be considered under Directive 90/220/EEC.
970. I conclude that the scientific evidence availa ble to Luxembourg in February 1997 was NOT
sufficient to permit it to undertake an appropriate assessment of potential risks to plants and the
environment from Event 176. Luxembourg could ha ve mustered sufficient evidence to argue that
there was an environmental risk that the target insects will evolve resistance to the Cry1Ab toxin,
resistance management measures should be required a nd resistance monitoring is a critical measure.
There was probably insufficient ev idence available to determine what kind of monitoring should be
required. This determination by L uxembourg was in direct contradiction to the EC decision. In this
case, Luxembourg probably does not need to claim th at their safeguard measure is a precautionary
one.
971. By 2003, the basis for risk assessment of Bt crops had changed because several significant
scientific points had come to light. (1) It became widely appreciated that the molecular basis of
transformation was more complex than originally th ought, and the implications of these findings for
risk assessment were articulated. Annex II of Directive 2001/18/EC is one consequence, but
additional regulatory changes have occurred since then. Indeed, knowledge in molecular biology
continues to accumulate at remarkably fast rates, an d I expect that there will be continued change in
regulation in the future. 132 (2) Non-target risk assessment shifted from assessing indicators of
environmental risk to assessing actual identified poten tial environmental risks. Presently this is done
on an ad hoc basis, as no systematic methodology h as gained widespread acceptance. (3) Gene flow
risk assessment has shifted from being based primarily on an assessment of the probability of gene
flow to being based on an assessment of both the probability of gene flow and the conditional hazard
probability. (4) Resistance risk is considered an environmental risk, and science-based resistance
management measures are required.
972. Thus, evidentiary standards for what constitut es an objective environmental risk assessment
had changed substantially from 1997 to 2003. This is particularly true in Europe and the United
States, and particularly true for the Bt crops.
973. (1) As in 1997, Luxembourg could muster sufficient evidence in 2003 to argue that there is an
environmental risk that the target insects will evolve resistance to the Cry1Ab toxin in Event 176,
resistance management measures should be required, and resistance monitoring should be required.
Moreover, in 2003, Luxembourg probably had sufficient evidence to determine what kind of
resistance management and monitoring measures to require. (2) Unlike in 1997, in 2003 Luxembourg
could argue that the molecular characterization of Event 176 is insufficient to conduct a risk
assessment. (3) Also in 2003, Luxembourg could argue that an additional gene flow risk that needs
assessment is contamination of conv entional production. It is possibl e that the scientific evidence to
support risk management measures for this risk were available to conduct an objective risk assessment
at that time.

131 Matten, S.R., et al., The U.S. Environmental Protection Agency's role in pesticide resistance
management, in Molecular Genetics and Evolution of Pesticide Resistance , T.M. Brown, Ed., American
Chemical Society, Washington, D.C., 1996, pp. 243-253.
132 Possibilities are suggested in Nap et al. 2004. Genomics for Biosafety in Plant Biotechnology . IOS
Press, NATO Science Series, Amsterdam.

Question 76: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Luxembourg evaluate the relevant risks to human, plant or animal health, and the
environment from the importation and use of Maize Bt-176?
(a) How does the scientific evidence a nd other documentation submitted by
Luxembourg compare with the relevan t international guidelines for risk
assessment and analysis identified above?
Dr. Nutti
974. Answer 76: Environmental issues are not my field, so I cannot judge this point, but all the
information for the safety assessment of potential risks to human and animal health that was submitted
by the applicants and evaluated by the Scientif ic Committee for Animal Nutrition, the Scientific
Committee for Food, the Scientific Committee for Pe sticides and the Scientific Committee for Plants
(SCP) were adequate and in accordance to the Codex Guidelines (FAO/WHO Codex Principles for
the Risk Analysis of Foods Derived from Modern Biotechnology; FAO/WHO Codex Guideline for
the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants;
FAO/WHO Codex Annex on Possible Allergenicity Assessment).
Dr. Andow
975. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks, specifically the resistance monitoring program.
976. With reference to the SPS Agreement, Luxem bourg has evaluated relevant risks to plant
health. However, it is not clear that Luxembourg has taken into account available scientific evidence
(Article 5.2). This is not to say that Luxem bourg has not taken into account available scientific
evidence.
977. It is not clear that Luxembourg conducted the risk assessment according to the phytosanitary
measures which might be applied. Luxembourg did not present their safeguard measure as a possible
phytosanitary measure. Luxembourg did not conduct the risk assessment according to their safeguard
measure. Thus it is not clear that Luxembourg c onducted the risk assessment consistent with Annex
A, paragraph 4.
978. It is not clear that Luxembourg took into account relevant economic factors (Article 5.3).
This is not to say that Luxembourg has not taken into account relevant economic factors.
979. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case because
Luxembourg argues that there is a plant pest risk.
980. For those risks within the scope of ISPM 11 the risk assessment process may be consistent
with ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
981. It appears that much of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of Luxembourg contradict this guidance. My reading of
the materials before the Panel is that Luxem bourg did not explicitly address this guidance.
Specifically I note Section 3, S1 (measures should be designed in proportion to the risk), Section 3.1

(level of acceptable risk should be expressed), Section 3.4 (principles), and Section 3.4.6 (on
prohibition) were not explicitly addressed.
982. The reasoning of Luxembourg is consistent with Annex III of the Cartagena Protocol on
Biosafety to the Convention on Biological Diversity.
Question 77: Does the scientific evidence and other information submitted by Luxembourg
support the adoption of a temporary prohibitio n on the importation and use of Maize Bt-176?
In light of any potential risks identified by Luxembourg, what other risk management options
were available in February 1997? What other risk management options are now available?
Dr. Nutti
983. It is important to consider that the main issu es raised by Luxembourg in order to prohibit the
use and sale of the Maize Bt-176 were that the GM product contains the gene conferring resistance to
ampicillin and that this circumstance risks having implication for human heal th, that in the present
state of the dossier cannot be properly evaluated and that no monitoring program has been
implemented with regard to the development of a resistance of the pyralid to the Bt-endotoxin. So,
these are environment related issues.
984. As far as my knowledge goes, regarding food safety and nutrition issues, the scientific
evidence submitted by Luxembourg does not support a temporary prohibition on the import and use of
Maize Bt-176.
Dr. Andow
985. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks, specifically the resistance monitoring program.
986. The adoption of a temporary prohibition can be justified on the basis of the scientific evidence
and other information available to Luxembourg. As there are no materials before the Panel containing
scientific evidence submitted by Luxembourg, it is not possible to make a determination of the
justification for the temporary prohibition based only on submissions from Luxembourg.
987. Other risk management options probably could not have been justified in February 1997.
However, Luxembourg could have proposed its own acceptable resistance monitoring measures,
which perhaps by 1999 would have been possible to implement.
988. Today several alternative risk management options are available for monitoring for resistance.
Although there is still no scientific consensus around the best monitoring method, the cost-efficiency
trade-offs are known, and consider able experience has accumulated so that several can be feasibly
implemented.

Maize MON 810 (notification C/F/95/12-02)
Safeguard measure of Austria
Question 78: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-159/At.1 and 2); and the Scientific Committee on Plants in February 1998
(CDA-82), September 1999 (US-55), and September 2000 (CDA-86), and the European
Commission in its Decision of April 1998 (CDA-81), as well as the information submitted by
Austria
 with respect to its safeguard measure (US-54, EC-159/At.4, EC-144, EC-147, EC-148), is
there any reason to believe that the scientific evidence available to Austria in June 1999 was
NOT sufficient to permit it to undertake an appropriate assessment of potential risks to human,
plant and animal health, and the environment from Maize MON 810? If so, what scientific
evidence do you believe was insufficient?
(a) If the evidence was not sufficient in June 1999, was there sufficient evidence
available to Austria in August 2003
 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from Maize MON 810 (EC-158/At.30-42)? If not, what scientific
evidence do you believe was insufficient?
Dr. Nutti
989. Answer 78: In EC 159-At 3, Austria informs the applicant that unt il the uncertainties which
have recently arisen during the review have been definitively clarified, Austria is justified in assuming
that the cultivation of the product MON810 presents a hazard to human health or the environment.
The main points presented were related to the undesired effects on non-target organisms, development
of resistance in insects, findings regarding the effects of other Bt plants (cotton in Australia and
USA). The document calls particular attention to the study carried out at Cornell University on the Bt
maize N4640Bt produced by Novartis, that sparked fierce reactions within the EU. The European
Commission, consequently, suspended the authorisation process for the herbicide-tolerant and insect-
resistant CryIA(b) Bt maize MON809 produced by Pioneer. Austrian CA also informs that no
conclusions have yet been reached by the Europ ean Commission regarding the Bt maize plants
MON810 produced by Monsanto and Bt-176 produced by Novartis, already approved for cultivation
in the EU.
990. The Scientific Committee on Plants (SCP) issued its opinion on the invocation by Austria of
Article 16 (safeguard clause) of Council Directiv e 90/220/EEc with respect to the placing on the
market of the GM maize MON 810, and the SCP conclusion was that the information submitted by
Austria did not constitute new significant information that was not already considered in its original
risk assessment and maintained the previous ri sk assessment status unchanged. The SCP also
concluded that this information does not invalidat e its original risk assessments for the other Bt
products. I believe the SCP has undertaken a very detailed analysis of the new information and its
remarks about the study carried out by Cornell are very detailed, explaining why that study was not
conclusive. I completely agree with the SCP opinion and conclusions.
991. Based on the information presented by notifiers and by the scientific evaluation undertaken by
France and by SCP, my understanding is there is no reason to believe that the scientific evidence
available to Austria in June 1999 was NOT suff icient to permit it to undertake an appropriate
assessment of potential risks to human, plant an d animal health, and the environment from the
importation and use of Maize MON 810.

992. Answer 78a: In my opinion, the scientific evidence presented was sufficient in 1999 and
2003.
Dr. Andow
993. My response to this question is the same as my response to question 72, except in the
following way. It is apparent from the Commission Decision (CDA-81, 22 April 1998) that resistance
management and monitoring measures have been proposed by the notifier. However, I could not find
in the materials before the Panel a full descripti on of these resistance management and monitoring
measures.
994. The Opinion of the SCP (CDA-82, 10 February 1998) does not provide a full description of
the proposed resistance management and monitoring measures.
995. Austria decided to prohibit the unregulated commercial use of Mon810 (EC-159/At.4, 10
June 1999). In its explanation for this action, Austria does not provide a full description of the
proposed resistance management and monitoring measures.
996. The further Opinion of the SCP (US-55, 24 September 1999) does not provide a full
description of the proposed resistance management and monitoring measures.
997. Rather than repeat the arguments in my response to question 72, I summarize as follows. I
conclude that the scientific evidence available to Austria in June 1999 was NOT sufficient to permit it
to undertake an appropriate assessment of potential risks to plants and the environment from Event
176. The evidence on risks to non-target organisms, including those in the soil could be considered
insufficient. I cannot provide a scientific judgment about the sufficiency of the scientific evidence on
resistance management and monitoring, because a full description of these measures is not available
in the materials before the Panel.
998. (1) In 2003, there remained some uncertainty about non-target risks of Mon810, but it could
be argued that a risk assessment could be conducted using scientifically justified worst-case
assumptions. On the other hand, Austria could reas onably maintain that there is still insufficient
information to know which non-target species may be at risk, and therefore it is not possible to
conduct an objective risk assessment. The findings in 2003 by Székács and Darvas (EC-158/At.37) –
that two protected butterfly species in Hungary, Inac his io and Vanessa atalanta, might be exposed to
Bt corn pollen and suffer higher mortality – certainly suggests that not all of the non-target species at
risk to Mon810 have been identified in Europe. However, the toxicity of Mon810 pollen had been
determined not to be high, so it is also possibl e that a worst case risk assessment would have found
insignificant risk even for these unknown species exposed to pollen. (2) Unlike in 1999, in 2003
Austria could argue that the molecular characterizati on of Event 176 is insufficient to conduct a risk
assessment. (3) In 2004, Austria sugge sted that an additional gene flow risk that needs assessment is
contamination of conventional production. It is po ssible that the scientific evidence to support risk
management measures is available today.

Question 79: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Austria evaluate the relevant risks to human, plant or animal health, and the
environment from Maize MON 810?
(a) How does the scientific evidence and oth er documentation submitted by Austria
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
999. Answer 79: Environmental issues are not my field, so I cannot judge this point, but all the
information for the safety assessment of potential risks to human and animal health that was submitted
by the applicants and evaluated by the lead CA (France) and the Scientific Committee for Plants
(SCP) were adequate and in accordance to the Codex Guidelines (FAO/WHO Codex Principles for
the Risk Analysis of Foods Derived from Modern Biotechnology; FAO/WHO Codex Guideline for
the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants;
FAO/WHO Codex Annex on Possible Allergenicity Assessment).
1000. The issues raised by Austria were environmental ones.
Dr. Andow
1001. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
1002. With reference to the SPS Agreement, Austria ha s evaluated relevant risks to plant health.
Specifically, Austria has taken into account available scientific evidence (Article 5.2).
1003. It is not clear that Austria conducted the risk assessment according to the phytosanitary
measures which might be applied. Austria did not present their safeguard measure as a possible
phytosanitary measure, and did not conduct the risk assessment according to their safeguard measure.
Thus it is not clear that Austria conducted the risk assessment consistent with Annex A, paragraph 4.
1004. It is not clear that Austria took into account relevant economic factors (Article 5.3). This is
not to say that it has not taken into account relevant economic factors.
1005. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case because
Austria argues that there is a plant pest risk.
1006. For those risks within the scope of ISPM 11 the risk assessment process is consistent with
ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
1007. It appears that much of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of Austria contradict this guidance. My reading of the
materials before the Panel is that Austria did not explicitly address this guidance. Specifically I note
Section 3, S1 (measures should be designed in proportion to the risk), Section 3.1 (level of acceptable
risk should be expressed), Section 3.4 (principle s), and Section 3.4.6 (on prohibition) were not
explicitly addressed.

1008. The reasoning of Austria is consistent with Annex III of the Cartagena Protocol on Biosafety
to the Convention on Biological Diversity.
Question 80: Does the scientific evidence and other information submitted by Austria support
the adoption of a temporary prohibition on Maize MON 810? In light of any potential risks
identified by Austria, what other risk management options were available in June 1999? What
other risk management options are now available?
Dr. Nutti
1009. Answer 80: The main issues raised by Austria in order to prohibit the use and sale of the
Maize MON 810 were related to the undesired e ffects on non-target organisms, development of
resistance in insects, findings regarding the effects of other Bt plants ( cotton in Australia and USA),
and these are environmental related issues.
1010. As far as my knowledge goes, concerning fo od safety and nutrition issues, the scientific
evidence submitted by Austria does not support a temporary prohibition of the import and use of
Maize MON 810.
Dr. Andow
1011. In my response to this question, I will not addr ess issues related to the antibiotic marker gene
and will focus my remarks on environmental risks.
1012. The adoption of a temporary prohibition can be justified on the basis of the scientific evidence
and other information submitted by Austria.
1013. Other risk management options may have been justified in June 1999. Because the full
resistance management and monitoring measures were not before the Panel, the scientific basis for a
concrete discussion of this is not possible.
1014. Today several alternative risk management options may be available. Risk management
strategies include risk avoidance, risk mitigation, and risk tolerance. Tolerance strategies are
probably inappropriate. Because the full resistan ce management and monitoring measures were not
before the Panel, the scientific basis for a concrete discussion of this is not possible.
Safeguard measure of Italy
Question 81: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-159/At.1 and 2); and the Scientific Committee on Plants in February 1998
(CDA-82), September 1999 (US-55), and September 2000 (CDA-86), and the European
Commission in its Decision of April 1998 (CDA-81), as well as the information submitted by
Italy
 with respect to its safeguard measure (EC-157/At.1 and 2, CDA-78), is there any reason to
believe that the scientific evidence available to Italy in August 1988 was NOT sufficient to
permit it to undertake an appropriate assessment of potential risks to human, plant and animal
health, and the environment from Maize MON 810 ? If so, what scientific evidence do you
believe was insufficient?
(a) If the evidence was not sufficient in August 1988, was there sufficient evidence
available to Italy in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the

environment from Maize MON 810? If not, what scientific evidence do you
believe was insufficient?
Dr. Nutti
1015. Answer 81: At EC 157-At.01, the Italian CA considers illegal the procedure for the placing
on the market of certain transgenic foodstuffs , namely BT11 MAIZE, MON810 MAIZE, and MS1
RF1, MS1 RF2 and GT-73 rapeseed oil, MON 809 MAIZE and T25 MAIZE, considering illegal
because the condition of "substan tial equivalence" to existing e quivalent foodstuffs required by
Article 3, Paragraph 4, of EC Regulation No 258/ 97 is not met. This document also points the
ambiguity of the term "substantial equivalence", that in the opinion of the Italian CA, this concept
could be defined differently, given the lack of cl arity in the Community legislation. The lead CA
informs that considering that the Superior Institute of Health, in its opinion, pointed out that, of the
seven notified products cited above, the four types of maize contain levels of protein deriving from
the genetic modifications ranging from 0.04 to 30 pa rts per million and that; therefore, the foodstuff
has been permanently affected by the modified elements. The lead Ca does not agree with the
technical documentation provided, as far as the co mparison of the GMOs and their conventional
counterpart is concerned. The document informs that the Superior Institute of Health also came to the
conclusion that, in the light of current scientific knowledge, there are no apparent risks to the health of
humans or livestock from the consumption of derivatives of the aforementioned GMOs, but the Italian
CA believes that there are inade quacies in the risk assessment procedures. The document also points
that the Superior Institute of Health declines to express an opinion regarding the risk of possible
"environmental release" of the GMOs in question.
1016. The Scientific Committee on Plants in February 1998 (CDA-82), after examining and
considering the existing information and data provide d in the dossier, considered that there was no
evidence to indicate that the seed s of GM maize MON810, when grown, imported and processed in
the manner indicated, are likely to cause adverse effects on human or animal health and the
environment.
1017. The Scientific Committee on Plants, in Febr uary September 1999 (US-55), concludes that
there is no new evidence that would change its previous assessment of GM maize MON 810.
1018. The Scientific Committee on Food, in Se ptember 2000 (CDA-86), presents its opinion
concerning the submission from the Italian authorities raising concerns from the safety of certain
products approved under the notification procedure of Regulation (EC) 258/97, concluding that the
information provided by the Italian Authorities does not provide detailed scientific grounds for
considering the use of the novel foods in question endangers human health.
1019. The European Commission Decision of 22 April 1998 gives consent for placing on the market
the inbred lines derived from maize line MON 81 0 and any progeny derived from crosses of the
product with any traditionally bred maize.
1020. My opinion is that the Italian CA did not provi de any scientific information that could change
the evaluations done by the scientific committees (Food and Plant). It is important to point out that the
information submitted by the applicant, covered all the items needed for the comparison of the GM
maize and its conventional counterpart, not only with data on nutrients but also with animal feeding
studies. The variation in levels of protein was in the range of natural variation, so the point raised by
Italy referring to protein was not correct.

1021. I do not agree with opinion of the Italian CA regarding the ambiguity of the term "substantial
equivalence", that, this concept could be define d differently, given the lack of clarity in the
Community legislation. The way to proceed fo r the comparison between the GM and non GM has
been presented not only at the Community legislati on but also at the OECD consensus documents,
FAO/WHO Expert Consultations 1996, 200, 2001 and Codex Alimentarius Guidelines.
1022. My understanding is there is NO reason to belie ve that the scientific evidence available to
Italy in August 1988 was NOT sufficient to perm it it to undertake an appropriate assessment of
potential risks to human, plant and animal health, and the environment from e of Maize MON 810.
1023. Answer 81a: In my opinion, the scientific evidence presented was sufficient in 1998 and
2003.
Question 82: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Italy evaluate the relevant risk s to human, plant or animal health, and the
environment from Maize MON 810?
(a) How does the scientific evidence and other documentation submitted by Italy
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
1024. Answer 82: All the information for the safet y assessment of potential risks to human and
animal health that was submitted by the applicants and evaluated by the lead CA (France) and the
Scientific Committee for Plants (SCP) and Scientific Committee for Foods were adequate and in
accordance to the Codex Guidelines (FAO/WHO Codex Principles for the Risk Analysis of Foods
Derived from Modern Biotechnology; FAO/WHO Code x Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible
Allergenicity Assessment).
1025. Answer 82a: The information and arguments pr ovided by Italy were not in accordance with
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants, paragraphs 44 , 45, 48.
Question 83: Does the scientific evidence and ot her information submitted by Italy support
the adoption of a temporary prohibition on Maize MON 810? In light of any potential risks
identified by Austria, what other risk management options were available in August 1988?
What other risk management options are now available?
Dr. Nutti
1026. Answer 83: The scientific evidence submitted by Italy do not support a temporary prohibition
of Maize MON 810.

Maize T25 (notification C/F/95/12-07)
Safeguard measure of Austria
Question 84: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-160/At.1 and 2); the SCP in September 2000 (CDA-75) and September 2001
(CDA-86 and CDA-77), and the European Commission in its Decision of April 1998 (CDA-74),
as well as the information submitted by Austria with respect to its safeguard measure (EC-
160/At.3 and 5, CDA-76, EC-144, EC-153), is there any reason to believe that the scientific
evidence available to Austria in April 2000 was NOT sufficient to permit it to undertake an
appropriate assessment of potential risks to human, plant and animal health, and the
environment from Maize T25? If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in Ap ril 2000, was there sufficient evidence
available to Austria in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from Maize T25 (EC-158/At.30-42)? If not, what scientific
evidence do you believe was insufficient?
Dr. Nutti
1027. Answer 84: In EC160 At.03, the Austrian CA pr ohibits the Maize T25, informing that this
event has not been examined under realistic cond itions of the use of this herbicide and of
correspondent agricultural practice. Neither the notification seeking approval of the placing on the
market T25 nor the decision of the European Co mmission is foreseeing a m onitoring programme. It
is emphasized that special measures for monitoring the possible spread of pollen to the fields in the
surrounding cultivated with conventional maize is missi ng, and the lack of this monitoring program is
crucial. The document also call attention to regiona l ecological aspects that are not differentiated and
the need for further investigations of eventual long term, also secondary, ecological effects.
1028. The SCP, in September 2000 (CDA-75), has considered that there was no evidence to indicate
that the use of the GM maize T25 as any other maiz e, is likely to cause adverse effects on human or
animal health and the environment.
1029. The SCP, in September 2001 (CDA-77), gave its opinion on the invocation by Austria of
Article 16 of Council Directive 99/22/EEC, regarding the GM maize T25, concluding that the
information submitted by Austria did not provide ne w scientific information to change the original
risk assessment carried out on the GM maize T25.
1030. The European Commission Decision of 22 April 1998 gives consent for placing on the market
the inbred lines derived from maize line T25 an d any progeny derived from crosses of the product
with any traditionally bred maize.
1031. Based on the information presented by the notifiers and by the scientific evaluation
undertaken by France and by the SCP, my understanding is there is NO reason to believe that the
scientific evidence available to Austria in June 19 99 was NOT sufficient to permit it to undertake an
appropriate assessment of potential risks to human, plant and animal health, and the environment from
the GM maize T25.
1032. Answer 84a: In my opinion, the scientific evidence presented was sufficient in 2000 and
2003.

Dr. Andow
1033. The reasons for Austria's safeguard measure are provided by Austria (CDA-76, EC-160/At.3,
20 April 2000). While the exact statement (unofficial translation) is reproduced below, there are 6
reasons stated. (1) The environmental risks of T25 have not been sufficiently evaluated under
realistic conditions. (2) There is no post-comme rcialization monitoring program. (3) Although harm
from pollen transfer to conventional maize production fi elds is likely absent, the potential for this risk
should be monitored. (4) There is no provision fo r the protection of ecologically sensitive regions.
(5) There is a need for regionally differentiated "good farming practice" guidelines to minimize the
danger of resistance. (6) There is a need to assess long-term and secondary ecological effects.
The maize line T25 had not been examined under realistic conditions of the use of
this herbicide and of correspondent agricu ltural practice. Neither the notification
seeking approval of the placing on the market of T25 nor the decision of the
European Commission are foreseeing a monitoring programme.
Furthermore special measures monitoring the possible – mostly regarded as safe –
spread of pollen to fields in the surroundi ngs cultivated with conventional maize are
missing.
The lack of a monitoring programme regarding long-term effects of genetically
modified plants or herbicides can be criticized especially because of the fact that the
approval conditions are not foreseeing a protection of sensitive areas (Hoppichler J.,
Expert Innenbefragung zur Bewertung und Evaluation, "GVO-freier ökologisch
sensibler Gebiete", Study on behalf of the Austrian Federal Chancellery, Vienna,
1999).
Furthermore regional ecological aspects are not differentiated: the use of herbicide
resistant plants in areas of unavoidable appli cations of herbicides seems to be useful,
if the good agricultural practice minimizes the danger of a resistance development.
Under other ecological – respectively agricu ltural – conditions the use of herbicide
resistant plants such as maize should only ta ke place after further investigations of
eventual long-term – also secondary – ecological effects.
1034. Regarding these reasons, (1) is possible grounds for the safeguard measure. (2) is a risk
management measure and must be justified by refe rence to a risk. Austria does not reference any
particular risk, so this cannot be grounds for the safeguard measure. (3) is a possible grounds for the
safeguard measure. (4) is a risk management measur e and must be justified by reference to a risk.
Austria does not reference any particular risk, so this cannot be grounds for the safeguard measure.
(5) Austria provides no argument that region-specifi c guidelines are necessary even when the risk of
resistance is regionally differentiated. Regarding th e risk of resistance, Austria has not differentiated
between the risk associated with volunteers and the risk associated with the evolution of resistance in
weeds. There is no scientific ground for resistance risk associated with maize volunteers in Europe.
Thus the only possible ground for the safeguard measure is the risk of resistance evolution in weeds.
(6) presents a class of risks that are possible gr ounds for the safeguard measure. Thus, I will
concentrate on the following four reasons – (1) The environmental risks of T25 have not been
sufficiently evaluated under realistic conditions ; (3) Although harm from pollen transfer to
conventional maize production fields is likely absent, the potential for this risk should be monitored;
the specified part of (5), viz., resistance risk in weeds, and (6) There is a need to assess long-term and
secondary ecological effects – as possible grounds for the safeguard measure.

1035. Regarding (6), long-term effects are certainly possible, but it is difficult to assess long-term
effects in pre-commercial risk assessments. In othe r words, risks that do manifest on long-term time
scales are difficult to predict, and unknown long-term effects must be managed after the fact. Thus,
the lack of long-term assessments of unknown effect s cannot be considered a reason for the safeguard
measure. As no long-term effects have been iden tified beyond resistance risk, this part of (6) cannot
be used to justify the safeguard measure. The remaining part of (6), secondary ecological effects, is a
specific case of reason (1) and will be treated together with (1).
1036. Regarding (3), Austria has acknowledged that th e possible harm is likely absent, but does not
specify concretely the possible harm. It is obvious that Austria cannot be thinking about
contamination of conventional produc tion (related to the "coexistence" issue) at this time. If it had
considered this, it would not acknowledge that the possible harm is likely absent. As the amended
SCP Opinion (CDA-77, 20 July 2001) also does not specific a possible harm, it is difficult to see how
this reason can justify the safeguard measure.
1037. Regarding (1), the amended SCP Opinion (CDA-77, 20 July 2001) does not provide an
assessment of environmental risks beyond gene flow risks. These would include non-target and other
biodiversity risks. However, Austria does not pr ovide any scientific eviden ce that such risks may
exist for T25 maize. A risk assessment cannot be co nsidered insufficient if all concrete possible risks
are addressed. In this respect (1) cannot be used to justify the safeguard measure.
1038. This leaves only resistance ri sk in weeds as the only possible grounds for the safeguard
measure. The amended SCP Opinion (CDA-77, 20 July 2001) does not provide an assessment of this
environmental risk. Even though Austria did not provided any scientific evidence that this risk exists
for T25 maize, resistance risks are widely recognized, and the consistent use of glufosinate with T25
maize would result in a resistance risk. Thus, there was insufficient scientific evidence available to
the SCP and Austria to assess weed resistance risk and appropriate risk management measures.
1039. Managing resistance risk in maize should be easi er than managing resistance risk in oilseed
rape, so I would expect that adequate information for a risk assessment could have been made
available by 2003, had the parties made concerted efforts to bring it together.
1040. However, as I have stated repeatedly, the evidentiary standards for what constitutes an
objective environmental risk assessment had changed substantially from 1997 to 2003. While this has
been particularly true for the Bt crops, there has also be a shift for GMHT crops. (1) It became widely
appreciated that the molecular basis of transforma tion was more complex than originally thought, and
the implications of these findings for risk asse ssment were articulated. Annex II of Directive
2001/18/EC is one consequence, but additional regulatory changes have occurred since then. Indeed,
knowledge in molecular biology contin ues to accumulate at remarkably fast rates, and I expect that
there will be continued change in regulation in the future.
133 (2) The UK-FSE trials have indicated
that there are possible risks to biodiversity associat ed with herbicide use on GMHT crops. However,
for GMHT maize, no adverse effects were found. (3) Contamination of conventional production
(related to the "coexisitence" issue) is a new issue that could be new grounds for the safeguard
measure. It is possible, however, that the scien tific evidence to support risk management measures
for this risk to conventional maize were available in 2003. Thus, in 2003, Austria could also argue
that T25 was inadequately characterized.

133 Possibilities are suggested in Nap et al. 2004. Genomics for Biosafety in Plant Biotechnology . IOS
Press, NATO Science Series, Amsterdam.

Question 85: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Austria evaluate the relevant risks to human, plant or animal health, and the
environment from Maize T25?
(a) How does the scientific evidence and oth er documentation submitted by Austria
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
1041. Answer 85: Environmental issues are not my field, so I cannot judge this point, but all the
information for the safety assessment of potential risks to human and animal health that was submitted
by the applicants and evaluated by the lead CA (France) and the Scientific Committee for Plants
(SCP) were adequate and in accordance to the Codex Guidelines (FAO/WHO Codex Principles for
the Risk Analysis of Foods Derived from Modern Biotechnology; FAO/WHO Codex Guideline for
the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants;
FAO/WHO Codex Annex on Possible Allergenicity Assessment).
1042. The issues raised by Austria are environment related.
Dr. Andow
1043. With reference to the SPS Agreement, Austria h as evaluated a risk relevant to plant health.
However, it is not clear that Austria has taken into account available scientific evidence (Article 5.2).
1044. It is not clear that Austria conducted the risk assessment according to the phytosanitary
measures which might be applied. Austria did not present their safeguard measure as a possible
phytosanitary measure, and did not conduct the risk assessment according to their safeguard measure.
Thus it is not clear that Austria conducted the risk assessment consistent with Annex A, paragraph 4.
1045. Austria did not take into account relevant economic factors (Article 5.3).
1046. The "potential pest" concept of ISPM 11 must first be considered. This is a special case of
the general argument in my response to question 6, and that argument holds for this case because
Austria argues that there is a plant pest risk.
1047. For those risks within the scope of ISPM 11 the risk assessment process is consistent with
ISPM 11. However, the economic assessments called for in ISPM 11 were not conducted.
1048. It appears that much of the ISPM 11 guidance on risk management has not been followed.
However, this is not to imply that the actions of Austria contradict this guidance. My reading of the
materials before the Panel is that Austria did not explicitly address this guidance. Specifically I note
Section 3, S1 (measures should be designed in proportion to the risk), Section 3.1 (level of acceptable
risk should be expressed), Section 3.4 (principle s), and Section 3.4.6 (on prohibition) were not
explicitly addressed.
1049. There is prima facie evidence that Austria did not follow Section 3, S1 (measures in
proportion to risk). Resistance risk a long-term ri sk. Austria probably could have phased in a series
of measures that would have allowed planting, but guar ded against resistance risk. It is not clear that
a prohibition is in proportion to the risk.

1050. The reasoning of Austria is probably consistent with Annex III of the Cartagena Protocol on
Biosafety to the Convention on Biological Divers ity. However, Austria provided little in science
documentation to conclude that this was consistent with Annex III.
Question 86: Does the scientific evidence and other information submitted by Austria support
the adoption of a temporary prohibition on Maize T25? In light of any potential risks identified
by Austria, what other risk management opti ons were available in April 2000? What other risk
management options are now available?
Dr. Nutti
1051. Answer 86: As far as my knowledge goes, re garding food safety and nutrition issues, the
scientific evidence submitted by Austria do not support a temporary prohibition on GM maize T25.
Dr. Andow
1052. The adoption of a temporary prohibition was probably not justified on the basis of the
scientific evidence and other information submitted by Austria. Austria needed to clarify the rationale
and provide substantial scientific evidence. A temporary prohibition probably could have been
justified at the time because the appropriate scientific information did exist.
1053. Several other risk management options could al so have been justified in April 2000. Risk
management strategies include risk avoidance, risk mitigation, and risk tolerance. In 2000 mitigation
and tolerance strategies were probabl y inappropriate. Here I consider only the resistance risk. One
risk avoidance strategy would have been to allow limited planting in a restricted region. This would
allow observing how T25 would be used and enable assessment of the selective pressure on weeds.
Another approach would have been to limit use on any particular field to once every 4-5 years. This
would reduce selection pressure for a long time, al lowing alternative management measures to be
developed.
1054. Today the same measures are available.
Safeguard measure of Italy
Question 87: Given the information before the Pa nel, including the evaluations undertaken by
France (EC-160/At.1 and 2); the SCP in September 2000 (CDA-75) and September 2001 (CDA-
86 and CDA-77), and the European Commission in its Decision of April 1998 (CDA-74), as well
as the information submitted by Italy
 with respect to its safeguard measure (CDA-78, EC-
157/At.1 and 2), is there any reason to believe that the scientific evidence available to Italy in
August 1988 was NOT sufficient to permit it to undertake an appropriate assessment of
potential risks to human, plant and animal health, and the environment from Maize T25? If so,
what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in August 1988, was there sufficient evidence
available to Italy in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from Maize T25? If not, what scientific evidence do you believe was
insufficient?

Dr. Nutti
1055. Answer 87: In EC 157-At.01 and CDA 78, the Italian CA considers illegal the procedure for
the placing on the market of certain transgenic foodstuffs, namely BT11 MAIZE, MON810 MAIZE,
and MS1 RF1, MS1 RF2 and GT-73 rapeseed oil, MON809 MAIZE and T25 MAIZE, considering
illegal because the condition of "substantial equivalence" to existing equivalent foodstuffs required by
Article 3, Paragraph 4, of EC Regulation No 258/ 97 is not met. This document also points the
ambiguity of the term "substantial equivalence", that in the opinion of the Italian CA, this concept
could be defined differently, given the lack of cl arity in the Community legislation. The lead CA
informs that considering that the Superior Institute of Health, in its opinion, pointed out that, of the
seven notified products cited above, the four types of maize contain levels of protein deriving from
the genetic modifications ranging from 0.04 to 30 pa rts per million and that; therefore, the foodstuff
has been permanently affected by the modified elements. The lead Ca does not agree with the
technical documentation provided, as far as the co mparison of the GMOs and their conventional
counterpart is concerned. The document informs that the Superior Institute of Health also came to the
conclusion that, in the light of current scientific knowledge, there are no apparent risks to the health of
humans or livestock from the consumption of derivatives of the aforementioned GMOs, but the Italian
CA believes that there are inadequacies in the risk assessment procedures. The document also points
out that the Superior Institute of Health declines to express an opinion regarding the risk of possible
"environmental release" of the GMOs in question.
1056. The SCP, in September 2000 (CDA-75), has considered that there was no evidence to indicate
that the use of the GM maize T25 as any other maiz e, is likely to cause adverse effects on human or
animal health and the environment.
1057. The Scientific Committee on Food, in Se ptember 2000 (CDA-86), presents its opinion
concerning the submission from the Italian authorities raising concerns from the safety of certain
products approved under the notification procedure of Regulation (EC) 258/97, concluding that the
information provided by the Italian Authorities does not provide detailed scientific grounds for
considering the use of the novel foods in question endangers human health.
1058. My opinion is that the Italian CA did not provi de any scientific information that could change
the evaluations done by the scientific committees (Food a nd Plant). It is important to point out that
the information submitted by the applicant covered all the items needed for the comparison of the GM
maize and its conventional counterpart, not only with data on nutrient but also with animal feeding
studies. The variation in levels of protein was in the range of natural variation, so the point raised by
Italy referring to protein was not correct.
1059. I do not agree with opinion of the Italian CA regarding the ambiguity of the term "substantial
equivalence", that this concept could be defined differently, given the lack of clarity in the
Community legislation. The way to proceed fo r the comparison between the GM and non GM has
been presented not only at the Community legislati on but also at the OECD consensus documents,
FAO/WHO Expert Consultations 1996, 200, 2001 and the Codex Alimentarius Guidelines.
1060. My understanding is there is NO reason to belie ve that the scientific evidence available to
Italy in August 1988 was NOT sufficient to perm it it to undertake an appropriate assessment of
potential risks to human, plant and animal health, and the environment from GM maize T25.
1061. Answer 87a: In my opinion, the scientific evidence presented was sufficient in 1998 and
2003.

Question 88: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Italy evaluate the relevant risk s to human, plant or animal health, and the
environment from Maize T25?
(a) How does the scientific evidence and other documentation submitted by Italy
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
1062. Answer 88: All the information for the safe ty assessment of potential risks to human and
animal health that was submitted by the applicants and evaluated by the lead CA (France) and the
Scientific Committee for Plants (SCP) and Scientific Committee for Foods were adequate and in
accordance to the Codex Guidelines (FAO/WHO Codex Principles for the Risk Analysis of Foods
Derived from Modern Biotechnology; FAO/WHO Code x Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible
Allergenicity Assessmen..)
1063. Answer 88a: The information and arguments pr ovided by Italy were not in accordance with
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants, paragraphs 44 , 45, 48.
Question 89: Does the scientific evidence and other information submitted by Italy support the
adoption of a temporary prohibition on Maize T25? In light of any potential risks identified by
Italy, what other risk management options were available in August 1988? What other risk
management options are now available?
Dr. Nutti
1064. Answer 89: The scientific evidence subm itted by Italy does not support a temporary
prohibition on GM Maize T25.
Maize MON 809 (notification C/F/95/12-01/B)
Safeguard measure of Italy
Question 90: Given the information before the Pa nel, including the evaluations undertaken by
the Scientific Committee on Plants in M ay 1998 and September 2000 (CDA-85 and CDA-86), as
well as the information submitted by Italy with respect to its safeguard measure (CDA-78, EC-
157/At.1 and 2), is there any reason to believe that the scientific evidence available to Italy in
August 1988 was NOT sufficient to permit it to undertake an appropriate assessment of
potential risks to human, plant and animal health, and the environment from Maize MON 809?
If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in August 1988, was there sufficient evidence
available to Italy in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from Maize MON 809? If not, what scientific evidence do you
believe was insufficient?

Dr. Nutti
1065. Answer 90: The SCP, in May 1998 (CDA-85), h as considered that there was no evidence to
indicate that the seeds of the GM MON809 and proge ny derived thereof, when grown, imported and
processed in the manner indicated, are likely to cause adverse effects on human and the environment.
1066. In EC 157-At.01 and CDA 78, the Italian CA c onsiders illegal the procedure for the placing
on the market of certain transgenic foodstuffs , namely BT11 MAIZE, MON810 MAIZE, and MS1
RF1, MS1 RF2 and GT-73 rapeseed oil, MON 809 MAIZE and T25 MAIZE, considering illegal
because the condition of "substan tial equivalence" to existing e quivalent foodstuffs required by
Article 3, Paragraph 4, of EC Regulation No 258/ 97 is not met. This document also points the
ambiguity of the term "substantial equivalence", that in the opinion of the Italian CA, this concept
could be defined differently, given the lack of clar ity in the Community legislation. The lead CA
informs that considering that the Superior Institute of Health, in its opinion, pointed out that, of the
seven notified products cited above, the four types of maize contain levels of protein deriving from
the genetic modifications ranging from 0.04 to 30 pa rts per million and that; therefore, the foodstuff
has been permanently affected by the modified elements. The lead Ca does not agree with the
technical documentation provided, as far as the co mparison of the GMOs and their conventional
counterpart is concerned. The document informs that the Superior Institute of Health also came to the
conclusion that, in the light of current scientific knowledge, there are no apparent risks to the health of
humans or livestock from the consumption of derivatives of the aforementioned GMOs, but the Italian
CA believes that there are inadequacies in the risk assessment procedures. The document also points
that the Superior Institute of Health declines to express an opinion regarding the risk of possible
"environmental release" of the GMOs in question.
1067. The Scientific Committee on Food, in Se ptember 2000 (CDA-86), presents its opinion
concerning the submission from the Italian authorities raising concerns from the safety of certain
products approved under the notification procedure of Regulation (EC) 258/97, concluding that the
information provided by the Italian Authorities does not provide detailed scientific grounds for
considering the use of the novel foods in question endangers human health.
1068. My opinion is that the Italian CA did not provi de any scientific information that could change
the evaluations done by the scientific committees (Food and Plant). It is important to point out that the
information submitted by the applicant, covered all the items needed for the comparison of the GM
maize and its conventional counterpart, not only with data on nutrient but also with animal feeding
studies. The variation in levels of protein was in the range of natural variation, so the point raised by
Italy referring to protein was not correct.
1069. I do not agree with the opinion of the Italian CA regarding the ambiguity of the term
"substantial equivalence", that this concept could be defined differently, given the lack of clarity in
the Community legislation. The way to proceed fo r the comparison between the GM and non GM has
been presented not only at the Community legislati on but also at the OECD consensus documents,
FAO/WHO Expert Consultations 1996, 200, 2001 and the Codex Alimentarius Guidelines.
1070. My understanding is there is NO reason to belie ve that the scientific evidence available to
Italy in August 1988 was NOT sufficient to perm it it to undertake an appropriate assessment of
potential risks to human, plant and animal health, and the environment from GM maize MON 809.
1071. Answer 90a: In my opinion, the scientific evidence presented was sufficient in 1998 and
2003.

Question 91: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Italy evaluate the relevant risk s to human, plant or animal health, and the
environment from Maize MON 809?
(a) How does the scientific evidence and other documentation submitted by Italy
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
1072. Answer 91: All the information for the safety assessment of potential risks to human and
animal health that was submitted by the applicants and evaluated by the lead CA (France) and the
Scientific Committee for Plants (SCP) and Scientific Committee for Foods were adequate and in
accordance to the Codex Guidelines (FAO/WHO Codex Principles for the Risk Analysis of Foods
Derived from Modern Biotechnology; FAO/WHO Code x Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible
Allergenicity Assessment).
1073. Answer 91a: The information and arguments pr ovided by Italy were not in accordance with
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants, paragraphs 44, 45, 48.
Question 92: Does the scientific evidence and other information submitted by Italy support the
adoption of a temporary prohibition on Maize MON 809? In light of any potential risks
identified by Italy, what other risk management options were available in August 1988? What
other risk management options are now available?
Dr. Nutti
1074. Answer 92: The scientific evidence submitted by Italy does not support a temporary
prohibition of GM Maize MON 809.
Maize Bt-11 (reference C/GB/96/M4/1)
Safeguard measure of Italy
Question 93: Given the information before the Pa nel, including the evaluations undertaken by
Great Britain (EC-163(At.1 to 3), the Scientific Committee on Plants in February 1998 (CDA-
83), November 2000 (CDA-84), and April 2002 (CDA-35-J), and the European Commission in
its Decision of April 1998 (CDA-80), as we ll as the information submitted by Italy with respect
to its safeguard measure (CDA-78, EC-157/At. 1 a nd 2), is there any reason to believe that the
scientific evidence available to Italy in August 1988 was NOT sufficient to permit it to undertake
an appropriate assessment of potential risks to human, plant and animal health, and the
environment from Maize Bt-11? If so, what scientific evidence do you believe was insufficient?
(a) If the evidence was not sufficient in August 1988, was there sufficient evidence
available to Italy in August 2003 to permit it to undertake a more objective
assessment of potential risks to huma n, plant and animal health, and the
environment from Maize Bt-11? If not, what scientific evidence do you believe
was insufficient?

Dr. Nutti
1075. Answer 93: The SCP, in February 1998 (CDA-83), has considered that the import of
genetically modified seed (notification C/GB/96/M4 /1) carrying the Bt11 event can be considered as
safe as utilising seed from non-genetically modified plants.
1076. In EC 157-At.01 and CDA 78, the Italian CA c onsiders illegal the procedure for the placing
on the market of certain transgenic foodstuffs , namely BT11 MAIZE, MON810 MAIZE, and MS1
RF1, MS1 RF2 and GT-73 rapeseed oil, MON 809 MAIZE and T25 MAIZE, considering illegal
because the condition of "substan tial equivalence" to existing e quivalent foodstuffs required by
Article 3, Paragraph 4, of EC Regulation No 258/ 97 is not met. This document also points the
ambiguity of the term "substantial equivalence", that in the opinion of the Italian CA, this concept
could be defined differently, given the lack of cl arity in the Community legislation. The lead CA
informs that considering that the Superior Institute of Health, in its opinion, pointed out that, of the
seven notified products cited above, the four types of maize contain levels of protein deriving from
the genetic modifications ranging from 0.04 to 30 pa rts per million and that; therefore, the foodstuff
has been permanently affected by the modified elements. The lead Ca does not agree with the
technical documentation provided, as far as the co mparison of the GMOs and their conventional
counterpart is concerned. The document informs that the Superior Institute of Health also came to the
conclusion that, in the light of current scientific knowledge, there are no apparent risks to the health of
humans or livestock from the consumption of derivatives of the aforementioned GMOs, but the Italian
CA believes that there are inade quacies in the risk assessment procedures. The document also points
that the Superior Institute of Health declines to express an opinion regarding the risk of possible
"environmental release" of the GMOs in question.
1077. The Scientific Committee on Food, in Se ptember 2000 (CDA-86), presents its opinion
concerning the submission from the Italian authorities raising concerns from the safety of certain
products approved under the notification procedure of Regulation (EC) 258/97, concluding that the
information provided by the Italian Authorities does not provide detailed scientific grounds for
considering the use of the novel foods in question endangers human health.
1078. The SCP, in November 2000 (CDA-84), considered that there was no evidence to indicate
that placing on the market for cultivation purposes of maize line Bt-11 and varieties derived from this
line by conventional crosses between Bt11 line and maize lines other than genetically modified ones,
was likely to cause any adverse effects on human health and the environment.
1079. The Scientific Committee on Food, in Apr il 2002 (CDA-35-J), presents its opinion on a
request to place on the market GM maize Bt-11, an d concludes that Bt-11 sweet maize is as safe for
human food use as its conventional counterparts. In this document, at item 3.4.2 – Compositional
Analysis, the Committee presents a detailed evaluati on of the data presented by the applicants and
concludes that the one case in which the content of crude protein of the transformed kernel was
significant lower than the isogenic control sample, no statistical differences have been observed and
concludes Bt-11 sweet maize kernels are substantia lly equivalent to a non-transformed lines except
for the new traits or proteins. In item 3.10 – Nutr itional Evaluation, the Committee concludes that the
Gm Bt-11, with respect to nutritional evaluation, is substantially equivale nt to non-modified maize
hybrids. In item 3.12.2 – Toxicological data on th e Cry1A(b) protein, the Committee refers to the
gastric and intestinal fluid tests, oral toxicologi cal tests and resistance to trypsin. Animal feeding
studies were also evaluated and no adverse effects were seen in these studies. I completely agree with
the Scientific Committee on Food, and my understandi ng is that the applicant has conducted all the
tests that were necessary for the safety assessment of the Bt-11.

1080. My opinion is that the Italian CA did not provi de any scientific information that could change
the evaluations done by the scientific committees (Food and Plant). It is important to point out that the
information submitted by the applicant covered all the items needed for the comparison of the GM
maize and its conventional counterpart, not only with data on nutrient but also with animal feeding
studies. The variation in levels of protein was not st atically significant, so the point raised by Italy
referring to protein was not correct.
1081. I do not agree with the opinion of the Italian CA regarding the ambiguity of the term
"substantial equivalence", that this concept could be defined differently, given the lack of clarity in
the Community legislation. The way to proceed fo r the comparison between the GM and non GM has
been presented not only at the Community legislati on but also at the OECD consensus documents,
FAO/WHO Expert Consultations 1996, 200, 2001 and the Codex Alimentarius Guidelines.
1082. My understanding is there is NO reason to belie ve that the scientific evidence available to
Italy in August 1988 was NOT sufficient to perm it it to undertake an appropriate assessment of
potential risks to human, plant and animal health, and the environment from GM maize Bt 11.
1083. Answer 93a: In my opinion, the scientific evidence presented was sufficient in 1998 and
2003.
Question 94: With reference to the definition of a risk assessment in the SPS Agreement (see
Background above), to what extent does the scientific evidence and other documentation
submitted by Italy evaluate the relevant risk s to human, plant or animal health, and the
environment from Maize Bt-11?
(a) How does the scientific evidence and other documentation submitted by Italy
compare with the relevant internationa l guidelines for risk assessment and
analysis identified above?
Dr. Nutti
1084. Answer 94: All the information for the safety assessment of potential risks to human and
animal health that was submitted by the applicants and evaluated by the lead CA (Great Britain) and
the Scientific Committee for Plants (SCP) and Scientific Committee for Foods were adequate and in
accordance to the Codex Guidelines (FAO/WHO Codex Principles for the Risk Analysis of Foods
Derived from Modern Biotechnology; FAO/WHO Code x Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants; FAO/WHO Codex Annex on Possible
Allergenicity Assessmet).)
1085. Answer 94a: The information and arguments pr ovided by Italy were not in accordance with
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants, paragraphs 44, 45, 48.
Question 95: Does the scientific evidence and other information submitted by Italy support the
adoption of a temporary prohibition on Maize Bt- 11? In light of any potential risks identified
by Italy, what other risk management options were available in August 1988? What other risk
management options are now available?

Dr. Nutti
1086. Answer 95: The scientific evidence submitted by Italy does not support a temporary
prohibition on GM Maize Bt-11.
ISSUE 3
Background: The following questions have the objective of assisting the Panel to determine
whether, for the specific biotech products at issue in this dispute, there are significant differences in
the risks arising to human, plant or animal health, or to the environment, from the consumption and
use of those products and:
(a) the consumption and use of those biotech products approved by th e European Communities
prior to October 1998, taking into account the scientific evidence available at the time of such
approvals;

(b) the consumption and use of comparable novel non-biotech products (s uch as plant products
produced by selective breeding, cross-breeding and mutagenesis) for consumption as food or
for planting or processing use; and

(c) the consumption and use of foods produced with biotech processing aids, including yeasts,
bacteria or enzymes that have been modified using recombinant DNA technology.

The experts are not being asked to judge the absolute significance of any differences, but to identify
whether differences exist and if so, to what extent, and to assist the Panel to understand the nature
and significance of any differences.

Furthermore, where there are no significant differen ces in the potential risks, the objective of the
questions is to assist the Panel to determine whether there are scientific or technical reasons for
managing any potential risks from the biotech products at issue in this dispute in a different manner
than other products (eg., monitoring requirements, agricultural management practices, etc.).

With regard to these questions, the experts may wish to comment on the replies by the Parties to the
questions by the Panel, in particular the rep lies to questions 23–26 and 29–32 (replies submitted
16 June 2004).

Questions:
Products of biotechnology approved by the European Communities prior to October 1998
Question 96: Has there been a significant chan ge in the understanding of physical, chemical
and biological characteristics of biotech products, gene interactions, gene expression, gene
silencing, molecular characterization, and product specific detection since 1998?
Dr. Nutti
1087. Answer 96: I will not judge any environmental issu e, since this is not my area of expertise.
However, in the areas of food safety, human and an imal nutrition, toxicology and allergenicity, the
same approach has been used since 1996 to assess the safety of GM products (FAO/WHO Expert
Consultation, 1996). The evolution of the safety assessm ent process in the last ten years is very much
related to the research (which generated a vast bibliography on the subject), discussion and public

perception/awareness developed during the peri od, when OECD produced several consensus
documents and the Codex Alimentarius approved the guidelines for the safety assessment of GM
plants and microorganisms. Also, a lot of resear ch has been conducted in different countries
(Australia, New Zealand, Japan, USA, European Union etc), in order to improve the whole process.
Nowadays, although we have in use (in all steps of the safety assessment) techniques which are more
refined, the approach remains essentially the same , fact which is clearly presented in the Codex
Alimentarius guidelines.
Dr. Squire
1088. From this reviewer's perspective, most of the underlying molecular and biochemical
knowledge of the subject has not changed substantia lly during this period. There is still uncertainty
over unintended side-effects occurring in biotech products, and the continued stability of phenotype in
biotech plants or continued expression of their gene s outwith the crop itself (i.e. in volunteers and
feral plants).
Question 97: On the basis of the information be fore the Panel, is there new scientific evidence
since 1998 that would suggest that the potential risks to human, plant or animal health, or to the
environment, from any of the specific biotech products subject to this dispute (including
products subject to the member State safegu ard measures), are different in nature or
magnitude as compared to the scientific understandi ng of the risks associated with such biotech
products prior to 1998, taking into account:
• the intended use of each product (direct human or animal consumption, further
processing for consumption, planting or any other specified use);
• any potential risks that may arise from the combination or successive use of
biotech products.
Does the information before the Panel support th e view that the potential risks from the
products in this dispute should be assessed differently than the risks from biotech products
approved prior to 1998?
Dr. Nutti
1089. Answer 97: My opinion is that, based on the existing literature and knowledge on the subject,
and from a scientific perspective, there is no new evidence that would suggest that the potential risks
to human, animal or plant health, from any of the sp ecific biotech products subject to this dispute, are
different in nature or magnitude as compared to those products of biotechnology approved by the
European Communities prior to October 1998.
1090. I will not judge any environmental issue, since this is not my area of expertise. However, in
the areas of food safety, human and animal nutrition, toxicology and allergenicity, the same approach
has been used since 1996 to assess the safety of GM products (FAO/WHO Expert Consultation,
1996). The evolution of the safety assessment process in the last ten years is very much related to the
research (which generated a vast bibliogr aphy on the subject), discussion and public
perception/awareness developed during the peri od, when OECD produced several consensus
documents and the Codex Alimentarius approved the guidelines for the safety assessment of GM
plants and microorganisms. Also, a lot of resear ch has been conducted in different countries
(Australia, New Zealand, Japan, USA, European Uni on etc), in order to improve the whole process.
Nowadays, although we have in use (in all steps of the safety assessment) techniques which are more

refined, the approach remains essentially the same , fact which is clearly presented in the Codex
Alimentarius guidelines.
1091. According to the WHO document "20 Questions on Genetically Modified Foods", available at
http://www.who.int/foodsafety/publications/biotech/en/20questions_en.pdf, the GM foods currently
available on the international market have passed risk assessments and are not likely to present risks
for human health. In addition, no effects on human health have been shown as a result of the
consumption of such foods by general population in the countries where they have been approved.
Continuous use of risk assessments based on the Codex principles and, where appropriate, including
post market monitoring, should form the basis for evaluating the safety of GM foods.
Dr. Andow
Changes since 1998:
1092. The main changes that have occurred since 1998 are related to risk assessment methodologies
and the evidence needed in a risk assessment. There are some di fferences between the transgenic
crops evaluated and approved before 1998 by the EC and those evaluated later, but these differences
are not as great as the changes in risk assessment methodologies and evidence needed. These changes
came about from increased scientific knowledge about transgenic crops. This has affected risk
assessment for transgenic crops intended for planting in the environment. Scientific investigation of
risks of combination biotech products has lagged behind.
1093. As I indicated in previous answers, the basi s for risk assessment of transgenic crops had
changed because several significant scientific points had come to light. (1) It became widely
appreciated that the molecular basis of transforma tion was more complex than originally thought, and
the implications of these findings for risk assessment we re articulated. (2) Non-target risk assessment
shifted from assessing indicators of environmental risk to assessing actual identified potential
environmental risks. Presently this is done on an ad hoc basis, as no systematic methodology has
gained widespread acceptance. (3 ) Gene flow risk assessment has shifted from being based primarily
on an assessment of the probability of gene flow to being based on an assessment of both the
probability of gene flow a nd the conditional hazard probability. (4 ) Resistance risk is considered an
environmental risk, and science-based resistance management measures are required.
1094. Thus, risk assessment methodologies and eviden tiary standards for what constitutes an
objective environmental risk assessment had cha nged substantially from 1998 to 2005. This is
particularly true in Europe and the United States, and particularly true for the Bt crops.
1095. Below, I will sketch some of the changes in non-target risk assessment and resistance risk
assessment and management.
Non-target risk assessment:
1096. Through 1997, most studies on non-target and biodi versity risks of transgenic plants showed
no effect of the transgenic plant (Fitt et al. 1994; Sims 1995; Dogan et al. 1996; Orr & Landis 1997;
Pilcher et al. 1997; Yu et al. 1997; EPA 2001; Monsanto Company 2002a, b). The methodological
approach used in nearly all of the studies wa s an indicator species approach similar to the
ecotoxicological assessments of pesticides where indi cator species are used to extrapolate to risks in
the actual environment. Only one laboratory study showed a lower survival of the springtail Folsomia
candida (Willem) when fed with high concentrations of Bt corn leaf protein (EPA 2001), but the
significance of this result is not clear. These indicat or species are not usually closely associated with

the transgenic plant tested or the area where the plants are grow n. Based on these studies, many
scientists believed that non-targets were not significantly at risk.
1097. In 1998, studies by Hilbeck et al (1998a,b) invigorated consideration of non-target risks by
reporting an unexpected adverse effect of Bt corn on the predatory green lacewing Chrysoperla carnea
Stephens. They fed C. carnea larvae with Cry1Ab Bt corn-fed prey or a diet containing purified
Cry1Ab toxin, and found higher immature mortality compared to controls. These results were
surprising because the Cry1Ab toxin is believed to be Lepidopteran-specific, while C. carnea belongs
to the Neuroptera, an order that is more closely related to the Coleoptera than to the mecopteroid
orders including the Lepidoptera. These results have been confirmed by additional studies (Hilbeck et
al. 1999; Dutton et al. 2002; Dutton et al. 2003), although the mechanism is still uncertain. These
studies suggested that Cry toxins may be less specific than previously believed.
1098. In the years following 1998, publications on no n-target risks began to shift from indicator
species to the actual (or tangible) potential risks on species that naturally occur in areas where
transgenic plants are meant to be cultivated. In early 1999, Losey et al. (1999) suggested that
monarch larvae (Danaus plexippus L.) suffered highe r mortality when feeding on their primary host
plant, the common milkweed Asclepias syriaca L., dus ted with transgenic Cry1Ab Bt pollen. This
initial observation was later confirmed by Jesse & Obrycki (2000) and coupled with the realization
that ~50% of the monarch breeding habitat is lo cated in the Corn Belt (Wassenaar & Hobson 1998),
this triggered concerns that large-scale cultivation of Bt corn would harm the monarch population.
Monarch butterflies attract wide interest in the US for multiple reasons, such as their beauty, iconic
significance to the public, and spectacular migration over several thousand miles. Stimulated by these
results, a group of researchers conducted a series of studies to estimate the actual risk of Bt corn to
monarch butterflies (Hellmich et al. 2001; Oberhauser et al. 2001; Pleasants et al. 2001; Sears et al.
2001; Stanley-Horn et al. 2001; Zangerl et al. 2001). Sears et al. (2001) concluded that the risk to
monarch populations was insignificant, and in an excellent review, Oberhauser & Rivers (2003)
summarized the events and finding s associated with these studies. Recent studies (Anderson et al.
2004; Jesse & Obrycki 2004; Dively et al. 2004), however, have revealed a higher toxicity of Bt
pollen and anthers than found in previous studies. Although Dively et al. (2004) suggested that the
risk to monarchs remains insignificant, a close anal ysis of the issues may allow other interpretations
of risk to monarchs.
1099. This shift to considering tangible risks may ha ve helped identify potential adverse effects to
the Federally endangered Karner blue butterfly (Lycaeides melissa samuelis Nabokov, Lepidoptera,
Lyceanidae). Instead of focusing only on commercial corn fiel ds, dispersal of pollen and the
production of corn in wildlife refuges could expose this endangered species to Bt pollen. The 2000
Scientific Advisory Panel (SAP) of the US E nvironmental Protection Ag ency (EPA) acknowledged
the possibility that this species ma y come in contact with Bt pollen (see also Andow et al. 1995), and
the EPA (2001) required additional assessment of the risks to Karner Blue butterfly.
1100. During 1999, research oriented toward assessing the tangible potential risks associated with
soils was published. Saxena et al. (1999) found th at Cry1Ab is released into the soil via corn root
exudates, where it can persist for at least 350 days (Saxena et al. 2002). These results suggested that
Bt corn could possibly affect rhizosphere and so il communities. Later, Zwahlen et al. (2003a)
reported that the Cry1Ab toxin in Bt corn litter pers isted for at least 8 months. Together these studies
showed that long-term exposure of soil organisms to Bt toxins was possible and that the risks of Bt
crops on soil ecosystem functioning should be assessed. Zwahlen et al. (2003b) also showed that
mortality and weight development of adult and juvenile earthworms, Lumbricus terrestris L., were not
significantly different when fed Bt or non-Bt corn residues, with the exception that after 200 days,
adult Bt corn-fed earthworms had a significant weight loss compared to the non-Bt corn-fed ones.

1101. Non-target risks associated with herbicide-to lerant crops were hardly studied until 2000,
when Watkinson et al. (2000) suggested that these cr ops might adversely effect skylark populations in
the UK. A large-scale field evaluation of herbicide-tolerant crops in the UK was established to
investigate possible actual effects on non-target speci es, and results were published in 2003. For the
most part, ecological effects propagated from whatev er changes in the weed community that resulted
from the change in herbicide use. The non-target e ffects of herbicide tolerant crops have not been
studied intensively elsewhere.
1102. In 2004, Andow & Hilbeck (2004) presented the outline of a new risk assessment model for
non-target effects of transgenic crops. Implicitly they are proposing to systematize the actual/
tangible risk assessment process th at has been building since 1999. An important innovation is to
select locally occurring non-target species that are mo st likely to be exposed to a transgenic crop and
likely to make significant contributions to the ecological functioning of the local ecosystem.
1103. Non-target and biodiversity risk assessments of tr ansgenic plants continue to be improved.
While indicator species continue to be used in many risk assessments, there is a trend towards
assessing actual/ tangible risks involving species that naturally occur in areas where transgenic crops
will be planted. In the future it will continue to be important to assess not only the effects of the
transgenic plant itself but also the effects associated with chang es in agricultural practices. In
addition, although they may be difficult to develop and verify, effective methods for biodiversity
assessment have not yet been developed.
Resistance risk assessment and management:
1104. The evolution of pest resistance to pest control measures has been known for nearly 100
years, but it became a significant problem after World War II, when modern, intensive agricultural
technologies proliferated, resulting in strong uniform selection over large areas. About 536 species of
arthropods, 60 genera of plant pathogenic fungi, and 174 weed species have evolved resistance to
pesticides (Eckert 1988; WeedScience.org 2003; Whalon et al. 2004), and resistance to Bt toxins has
been documented in >17 insect species (Tabashnik 1994, Huang et al. 1999). Interestingly,
virologists remain unconvinced that resistance will evolve to transgenic virus resistant crops (Tepfer
2002), however, despite some disagreement (reviewed in Tabashnik 1994), entomologists and weed
scientists agree that resistance evolution is a real risk for which some management is desirable
(NRC 1986).
1105. At the beginning of the 1990s, it had proven di fficult to implement effective resistance
management for most pesticides. Indeed, there was pessimism that a high-dose/ refuge resistance
management system could ever be implemented for insecticide resistance management because a high
dose could not be reliably maintained (Roush 1989; Tabashnik 1989). With the advent of transgenic
Bt crops, hopes were renewed (Gould 1994; Roush 1994) , but it was not clear that a high-dose/ refuge
strategy would delay resistance enough with a reasonably sized refuge (Comins 1977).
1106. In a series of simple simulations based on Comins (1977) early work, Alstad & Andow
(1995) showed that the high-dose/ refuge strategy c ould delay resistance in European corn borer to Bt
corn for more than 30 years with a 50% non-Bt co rn refuge. Subsequent research suggested that
smaller refuges would also substantially dela y resistance, proving that effective resistance
management was possible theoretically (Gould 1998; Shelton et al. 2000).
1107. The focus shifted to practicalities. Could an effective resistance management strategy be
implemented? In the US, this question was answer ed through a series of decisions made by the EPA.
In early 1995, the EPA registered Bt potato, and although resistance risk was recognized, no

resistance management was required. By the end of that year, EPA issued conditional registrations
and required the development of resistance manageme nt for all subsequent Bt crops (Matten et al.
1996). Conditional registrations were used to mo tivate the development and implementation of a
scientifically justified resistance management strategy.
1108. The key issues in 1995 were how large a refuge was needed, did the refuge need to be
spatially structured relative to the Bt fields, and could the refuge be managed to limit pest losses?
Aspects of some of these questions remain unresolved today. In Australia, Bt cotton did not provide a
high-dose against the key pest, cotton bollworm Helicoverpa armigera (Hübner), and growers and
researchers agreed to require 70% refuges to make the likelihood of resistance remote (Fitt 1997). In
the US, Bt cotton provided a high-dose against the key pest, cotton budworm Heliothis virescens (F.),
but not against another important pest, Helicoverp a zea (Boddie), however, refuge requirements were
set at 4% unsprayed or 20% sprayed refuge outside of the Bt field with minimal requirements of
spatial structure. In 2001, spatial structure requi rements were added for Bt cotton along with other
modifications. These initial requirements and changes represent a compromise among various
interests, although science played a significant role.
1109. Resistance management requirements for Bt corn developed with strong scientific input.
Early in 1997, the USDA regional research committee NC-205 reviewed model results and
information on the ecology of European corn borer and suggested to registra nts and the EPA that a
20-25% refuge was needed near all Bt corn fiel ds (Anon. 1998). Research results supporting this
recommendation were published in the ensuing y ears (Onstad & Gould 1998; Hunt et al. 2001;
Bourguet et al. 2003). One of the key results was a bioeconomic model suggesting that a 20% refuge
would be nearly optimal for growers who consider the trade-off between the immediate costs of the
refuge and delayed costs of resistance failures (H urley et al. 2001). Canada required a 20% refuge
within 0.5 miles (~800 meters) of Bt corn in 1998, and during 1999 a consensus was reached in the
US and the EPA required a 20% refuge within 0.5 miles of Bt corn for the 2000 growing season and
thereafter.
1110. Several scientific issues remain unresolved. Understanding the mechanisms of resistance is
necessary to tailor resistance management to the par ticular system, but these are just beginning to be
revealed for Bt crops (Gahan et al. 2001). The details of adult movement may play a key role in the
evolution of resistance (Caprio 2001; Ives & Andow 2002). Limited dispersal of adults from natal
fields (Comins 1977), pre- versus post-mating adult movement, and male versus female movement
(Ives & Andow 2002) may have significant effects on m odels of resistance evolution, and estimating
these movement rates in the field is challenging. Farming practices, such as crop rotation (Peck &
Ellner 1997), management of the refuge (Onsta d et al. 2002; Ives & Andow 2002), and rational
approaches to pest management, may affect the rates of resistance evolution. Significantly, a
consensus for managing low-dose events has yet to emerge, and scientific analysis of this problem is
incomplete. For example, Australia implemented 70% refuges for one low-dose event, while the US
has used 20% refuges for both high- and low-dose events.
1111. Monitoring for the occurrence and frequency of resistance and methods to improve
compliance to resistance management requirements among growers are areas of current research. The
key monitoring problem is how to estimate resistance when it is rare and recessive. One promising
approach is the F
2 screen (Andow & Alstad 1998; Andow & Ives 2002; Stodola & Andow 2004). It is
a genic screen and works by inbreeding isofemale lines so that recessive phenotypes are expressed in
the F 2 generation, when they can be screened. If mated females are collected from natural
populations, each carries four haplotypes (two of her own and two of her mate's) and only 250 female
lines need to be screened instead of 10 6 field-collected individuals. The F 2 screen has been used for
several species (Bentur et al. 2000, Bourguet et al. 2003; Génissel et al. 2003). Other genetic and

phenotypic methods have been used on some cotton pests (Gould et al. 1997; Tabashnik et al. 2000),
but usually phenotypic screens will have lower sensi tivity and higher cost than genic screens (Andow
& Ives 2002). Improving compliance will require a combination of bioeconomic modeling, surveys
of grower behavior and motivations, and development of effective educational materials.
1112. Our understanding of resistance risk and management continues to evolve. Presently, none of
the Bt crops now used has suffered a resistance failure despite widespread use. While this may be due
to good fortune, in some cases, such as Bt cotton in Australia, resistance management must have been
crucial to avoiding failure, and in other cases, such as Bt cotton in Arizona, US, other factors must
also be important (Tabashnik et al. 2003). Interes tingly, relatively little research has focused on weed
resistance to herbicides used with the herbicide tolerant crops. This problem has been treated as a
theoretical herbicide resistance problem (Gressell et al. 1996), but with recent reports of weed
resistance (WeedScience.org 2003), this may change.
134

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Dr. Snow
1113. I conclude that yes, new scientific evidence has become available since 1998 with regard to
gene flow from herbicide-tolerant crops and taking into account the environmental concerns and
management goals of EC Member States. New scientific studies published during 1998-2003 showed
that the dispersal of transgenes that confer resi stance to glufosinate and glyphosate will occur much
more widely and more quickly than was previously expected, as I discuss in Part I. Also, new studies
showed that the widespread use of glufosinate-tolerant and glyphosate-tolerant oilseed rape could lead
to populations of volunteers and weeds that are mo re difficult to manage than their nontransgenic

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predecessors. Adopting these herbicide-tolerant cr ops could lead to greater dependence on these and
other herbicides (see Part I and Friesen et al. 2003).
1114. These problems would not arise without the use of transgenic crops b ecause the genes that
confer resistance to glufosinate and glyphosate have not been found to occur naturally in the crop's
gene pool. Although another variety of oilseed rape with resistance to the herbicide imidazolinone is
widely grown in Canada and Australia, the relativel y minor problems associated with its use could be
compounded with the addition of transgenes that provide more resistance to more herbicide modes of
action. The Canadian regulatory system treats al l herbicide-tolerant crops in a similar manner,
regardless of whether they are transgenic or not, and it appears that little or no effort has been made to
avoid gene flow among herbicide- tolerant crops, volunteers, and w eeds. The EC approach differs
dramatically in that EC considers possible long-term effects that might arise from the combination or
successive use of herbicide-tolerant crops.
1115. Application rates of glufosinate and glyphosate are expected to increase greatly if these
herbicide-tolerant crops are adopted by farmers. In Europe, scientists are still investigating the
question of whether these types of changes in ag ricultural practices could have unwanted effects on
farmland biodiversity, relative to ongoing effects of c onventional, nontransgenic crops. This research
started during 1998-2003 and findings have recently begun to appear in scientific publications (e.g.,
Watkinson 2000, Firbank 2003, Squire et al. 2003). Also, in the context of Europe, gene flow
between transgenic oilseed rape crops and non-transg enic crops is perceived as an economic problem
due to newly introduced labelling requirements. Research is under way to develop effective strategies
for "coexistence" of transgenic and conventional crops.
1116. As discussed above, scientific experts in th e EC have concluded that potential problems
related to gene flow should be monitored after commercialisation, and that farmers should be
educated about appropriate stewardship and manageme nt of herbicide-tolerant crops. The scientific
committees of certain Member States (e.g., France) have argued that more research is needed to
determine whether problems related to gene flow can be mitigated or avoided after these crops are
grown on a large scale. In Canada and the US, how ever, no measures have been taken to avoid these
problems. Glyphosate-tolerant oilseed rape is gr own very widely in western Canada, where this
herbicide is used in low-till and no-till weed management.
1117. Here, I have focused on herbicide-tolerant crops . Other scientific studies have raised valid
questions about the persistence of Bt toxins in the environment and their potential to harm nontarget
insects (e.g., Losey 1999, Stotzky 2001, Hilbeck 2001). To my knowledge, these questions largely
have been resolved by subsequent scientific findings with regard to th e crops that are the focus of this
dispute.
1118. Furthermore, I do not know of any studies or scientific evidence showing that the process of
creating transgenic plants is inherently more risky than other methods used in conventional breeding,
including mutagensis and embryo rescue following wide crosses.
1119. In summary, the types of new scientific informa tion that I discuss in Part I of my answers can
be used to support the view that potential risks from the products in this dispute could be assessed
differently than the risks from biotech products approved prior to 1998.
Dr. Squire
1120. The argument presented in Notes 11-12 is that knowledge of the ecological impacts of biotech
products and their spread and concentration in the agricultural environment has changed, particularly

in that emergent properties at the scales of the field and landscape are now much better appreciated
even if they are still far from fully underst ood. Examples of where knowledge has increased
substantively are –
• Spread and persistence of some crops plants, notably oilseed rape.
• The important role of insects as carriers of pollen over the landscape.
• The importance of the weed flora to in-field biodiversity and food webs.
Question 98: From a scientific perspective, is there a significant difference in risks to human,
animal or plant health or the environment ar ising from the use of a bacterial antibiotic
resistance marker gene, or part thereof, in any biotech product at issue in this dispute (eg,
Monsanto Bt cotton (531), Monsanto Ro undup Ready cotton (RRC1445), Amylogene starch
potato) compared to those products of biotec hnology approved by the European Communities
prior to October 1998?
Dr. Nutti
1121. Answer 98: As I already answered in question 1, it will be necessary a combination of several
factors for a DNA transfer from plants to microbial or mammal cells to occur, which is the main
argument raised in this dispute against the products using antibiotic resistance marker genes.
1122. It is important to point out that according to FAO/WHO (2000), DNA transfer from plants to
microbial or mammal cells, under normal circumstances of dietary exposure, would require all the
following events to occur:
• the relevant gene(s) would have to be released, probably as a linear fragment;
• the gene(s) would have to survive nucleases in the plant and in the gastrointestinal
tract;
• the gene(s) would have to compete for uptake with dietary DNA;
• the recipient bacteria or mammalian cells would have to be competent for
transformation and the gene(s) would have to survive their restriction enzymes and
• the gene(s) would have to be inserted in the host DNA by rare repair or recombinant
events.
1123. There have been numerous experiments aimed at evaluating the possibility of transfer of plant
DNA to microbes and mammalian cells. To date, there are no reports that marker genes in plant
DNA transfer to these cells. Even so, the use of alternative transformation methods, which do
not use ARMG, is encouraged. If alternative marker genes are used, they also must be evaluated
regarding their safety.
1124. The transfer of marker genes which confer resistance to kanamycin, ampicillin and
streptomycin to bacteria in the human gut is unlikely to present a significant health impact since
bacteria resistant to these antibioti cs are already spread all over or are naturally found in the human
gastrointestinal tract (S malla et al., 1993; Calva et al., 1996; Shaw et al., 1993; Smalla et al., 1997).

Besides, kanamycin/neomycin and streptomycin ar e rarely used for humans due to their collateral
effects (WHO 1993).
1125. My opinion is that, based on the existing lite rature and knowledge on the subject, and from a
scientific perspective, there is no new evidence that would suggest that the potential risks to human,
animal or plant health arising from the use of a b acterial antibiotic resistance marker gene, or part
thereof, from any biotech product at issue in this dispute (eg, Monsanto Bt cotton – 531, Monsanto
Roundup Ready cotton – RRC1445, Amylogene starch potat o) are different in nature or magnitude as
compared to those products of biotechnology a pproved by the European Communities prior to
October 1998.
1126. I will not comment on any environmental issue, since this is not my area of expertise.
However, in the areas of food safety, human and an imal nutrition, toxicology and allergenicity, the
same approach has been used since 1996 to assess the safety of GM products (FAO/WHO Expert
Consultation, 1996). The evolution of the safety assessm ent process in the last ten years is very much
related to the research (which generated a vast bibliography on the subject), discussion and public
perception/awareness developed during the peri od, when OECD produced several consensus
documents and the Codex Alimentarius approved the guidelines for the safety assessment of GM
plants and microorganisms. Also, a lot of resear ch has been conducted in different countries
(Australia, New Zealand, Japan, USA, European Union etc), in order to improve the whole process.
Nowadays, although we have in use (in all steps of the safety assessment) techniques which are more
refined, the approach remains essentially the same.
135
Question 99: For those biotech products at issue in this dispute for which no significantly
different nature or level of risk has been id entified, does the information before the Panel
provide a scientific or technical rationale for monitoring the occurrence of potential adverse
effects, or of unintentional effects, arising from the consumption or use of these products
compared to those products of biotechnology ap proved by the European Communities prior to
October 1998?
Dr. Nutti
1127. Answer 99: In my opinion, the biotech products at issue in this dispute should be treated in
the same way as the biotech products approved by the European Communities prior to 1998.

135 References:
FAO/WHO. Safety aspects of genetically modified foods of plant origin. Report of a Joint FAO/WHO
Expert Consultation on Food Derived from Biotechnology, Geneva, Switzerland, 29 May-2 June 2000. Rome:
Food and Agriculture Organization of the United Nations. 2000. www.fao.org/es/esn/gm/biotec-e.htm.
CALVA, J.J.; SIFUENTES-OSBORNIO, J.; CERON, C. Antimicrobial resistance in fecal flora:
longitudinal community-based surveillance of children from urban Mexico. Antimicrob. Agents Chemother.,
vol. 40, pp. 1699-1701, 1996.
SHAW, K.J., RATHER, P.N., HARE, R.S. AND MILLER, G.H. Molecular genetics of
aminoglycoside resistances genes and familial relationships of the aminoglycosidemodifying enzymes.
Microbiol. Rev., vol. 57, pp. 138-163, 1993.
SMALLA, K.; VAN OVERBEEK, L.S.; PUKALL, R.; VAN ELSAS, J.D. Prevalence of nptII and Tn5
in kanamycin resistant bacteria from different environments. FEMS Microbiol. Ecol., vol. 13, pp. 47-58, 1993.
SMALLA, K.; WELLINGTON, E.; VAN ELSAS, J.D. Natural background of bacterial antibiotic
resistance genes in the environment. Nordic Seminar on Antibiotic Resistance Mark er Genes and Transgenic
Plants, p 8-16. June 12-13, 1997, Oslo, Norway. The Norwegian Biotechnology Advisory Board.
WHO (1993). Health aspects of marker genes in genetically modified plants, Report of a WHO
Workshop. World Health Organisation, Geneva.

1128. The products approved prior to 1998 are the so called "first generation" products. Most of
them had the new trait introduced in order to become insect resistant or herbicide tolerant, and they
were assessed as equivalent to their conventional counterparts. The products approved prior to
October 1998 have been consumed in different regions of the wo rld (USA, Canada, Argentina,
Australia, New Zealand) and, no adverse effects re lated to their consumption have been reported in
any of these countries.
1129. According to the WHO document "20 Questions on Genetically Modified Foods", available at
http://www.who.int/foodsafety/publications/biotech/en/20questions_en.pdf, the GM foods currently
available on the international market have passed risk assessments and are not likely to present risks
for human health. In addition, no effects on hu man health have been shown as a result of the
consumption of such foods by general population in the countries where they have been approved.
Continuous use of risk assessments based on the Codex principles and, where appropriate, including
post market monitoring, should form the basis for evaluating the safety of GM foods.
Dr. Andow
1130. There are some biotech products at issue in th is dispute for which one committee or another
has determined that were no significant risks. I have taken issue with some of these determinations in
my previous responses. For all of the products at issue in this dispute, there are some member
countries that disagree with the assessment of the relevant SCP (for example) that there are no
significant identified risks. This disagreement is sometimes related to different standards of
acceptable risk being applied by the disagreeing parties.
1131. This disagreement provides the only rationale for monitoring potential adverse effects. If all
parties agreed that there were no significant iden tified risks, there would no need to consider
monitoring for potential adverse effects.
1132. Unanticipated effects could arise from a transg ene by any of the following mechanisms: new
ORFs, insertional mutagenesis, post-transcriptional or post-translational processing, other pleiotropy
or epistasis, and gene-by-environment interaction. New ORFs and insertional mutagenesis can be
addressed through molecular characterization of all transgene loci. Post-transcriptional or post-
translational processing can be addressed to the most part by molecular and biochemical analysis.
Other pleiotropy or epistasis and gene-by-envi ronment interaction cannot be addressed without
extensive planting in the field.
1133. All of these possible sources unanticipated effects are equally likely to occur for plant biotech
products before 1998 and plant biotech products after 1998. It is possible that more recent events
(after 2001) are less likely to have such unanticip ated effects than those prior to 1998. The main
difference between them is that new ORFs, inser tional mutagenesis and post-transcriptional and post-
translational processing was not looked for very much before 1998, while after 2001, the methods and
standards have become increasingly targeted to assess these possibilities. This provides a rationale for
monitoring.
Dr. Snow
1134. This question refers to products for which no si gnificant risks have been identified, whereas
many of the questions above refer to products fo r which one or more Member States has identified
potential risks.

1135. With regard to environmental effects, if this question pertains only to products that have no
identified environmental risks, then there is no sci entific rationale for obligatory monitoring. One
could argue that possible risks may have been overlooked prior to marketing, but it is not logical to
require monitoring for every transgenic crop, regardl ess of its phenotypic traits. Rather, this might be
carried out to reassure the public that long-term studies are being conducted, or to ensure that
labelling thresholds for non-GM products are not exceeded.
1136. If gene flow from transgenic crops is viewed as a form of genetic "contamination", then
determining the extent to which it occurs after deregulation could be seen as a socio-economic reason
for monitoring. In this context, scientific knowledge that was gained after 1998 is relevant because
many studies showed that gene flow occurs more widely and more quickly than was previously
anticipated (see above).
1137. In reality, the scientific basis for which envi ronmental consequences to monitor and how to
do this in a scientifically rigorous manner are not well developed (e.g., NRC 2002). There are certain
types of environmental effects for which monitori ng can be justified on scientific or technical
grounds. Specific and plausible environmental harm s should be identified before allocating time and
money to these efforts.
Question 100: For those biotech products at is sue in this dispute for which an approval has
been sought for environmental release (notif ications submitted under Directives 90/220 or
2001/18), and for which no significantly different nature or level of risk has been identified, does
the information before the Panel provide a scientifi c or technical rationale for requiring specific
agricultural management practices that differ from those for products of biotechnology
approved by the European Communities prior to October 1998?
Dr. Andow
1138. A parallel argument to my response to question 99 holds for this question.
Dr. Snow
1139. See Answer 99 because the same principles apply.
Dr. Squire
1140. Following the points made in answer to Q.97, the innate biological qualities and behaviour of
a product may not have changed, but it is quite possi ble that the context in which that product would
operate has changed, in two ways. (A) The physic al or agronomic environment is different. The
agronomy of these crops changes rapidly: examples of factors that have changed markedly in Europe
in recent times are the proportion of autumn-sown crops, the area covered by oilseed rape, and the
type and effectiveness of pesticides used. An example, given already, of the change in pesticides is the
rise in use of glyphosate as an arable herbicide in some countries in the past few years. (B) The
understanding of the GM product's role in the ecosyst em has changed. It was also reported previously
that some major effects of intensification on arable ecosystems have become well established
scientifically only in the last few years. The way ar able plants and their food webs are considered is
now different from the way they were generally considered in the early 1990s. The question is
sometimes referenced back to before the early 1990s: why was the large scale change to autumn-sown
cropping in some parts of Europe not scrutinised w ith the same rigour as the potential change to GM
cropping? The answer is that it was thought not to be important, while it is now known to have been
very important for farmland food webs.

Question 101: Does the information before the Pa nel support the argument that any potential
risks from any of the biotech products at issue in this dispute should be mitigated in a manner
different than the products of biotechnology approved by the European Communities prior to
October 1998? If so, what means of risk mitigation might be envisaged?
Dr. Andow
1141. None of the products approved by the EC prior to October 1998 have been mitigated in any
manner by the EC because no mitigable risks have been reported and verified. Should any of the
biotech products cause a verifiable risk, this would be sufficient argument for mitigating them
differently.
1142. However, as indicated in my response to question 97, there are new risk methodologies and
assessment standards being applied to biotech products today than prior to 1998. Thus, it is possible
that risks will be identified for new products that we re not even considered in the older products.
Under such conditions, differences could be justified.
Dr. Snow
1143. Several of the products that we re approved before 1998 are the same as those in this dispute
or are very similar. These include the following, as listed in the US June 16, 2004 documents:
Oilseed rape (= Swede rape)
Glufosinate-tolerant, male sterile (MS1,RF1); C/UK/94/M1/1
Glufosinate-tolerant (Topas 19/2); C/UK/95/M5/1
Maize
Bt-176, glufosinate-tolerant; Ceiba-Geigy; C/F/94/11-03
Bt cry1A; MON 810; Monsanto; C/F/95/12-02
Bt-11; glufosinate-tolerant with Bt cry1Ab; C/UK/96/M4/1
Glufosinate-tolerant (T25); AgrEvo; C/F/95/12-07
Chicory
Glufosinate-tolerant, male sterile; C/NL/94/25
1144. The answer to this question hinges on whic h potential environmental risks might require
mitigation, and whether any new scientific knowledge has become available to justify changes to any
mitigation plans that were required prior to October 1998.
1145. In summary, I am not sure which mitigation plans may have been required at that time, but
new scientific knowledge that was gained after 1998 cer tainly could be relevant to risks related to
gene flow (see above) and the evolution of resistance in target pests of Bt crops (e.g., NRC 2000).
1146. In the case of oilseed rape that was approved for import and processing only, I cannot identify
new scientific reasons for different mitigation plan s (e.g., using covered vehicles when transporting
large quantities of seeds).
Dr. Squire
1147. The answers follow from the responses above to questions 97 and 100. While products might
have very similar qualities, mitigation might differ as a result of new information informing risk or a
new perception of the importance of a particul ar risk. Among ecological topics, the perceived
mitigation to reduce the 'severity' of a risk to biodiversity and ecosystem functioning has changed

because of a greater appreciation that arable syst ems have been affected by intense agriculture.
Mitigation is unlikely to remain constant. For example, some current mitigation measures which leave
margins round the cropped area of land are insufficient when it is appreciated th at biodiversity in the
cropped areas is necessary to main ecological function there.
1148. That risk has not remained the same over tim e is an inevitable consequence of scientific
information on arable systems being collected at increasingly larger scales since the mid-1990s. Early
research using GMOs in small parcels of land coul d not possibly have generated the knowledge that
pollen from large fields of oilseed rape is move d widely in the landscape. Moreover, the present
guidelines about pollen barriers, e.g. a strip of flowering crop between the GM field and non-GM
recipient field, may have to change if the proportion of GM fields in the landscape increased to, say,
50%.
Comparable novel non-biotech products, (such as plant products produced by selective
breeding, cross-breeding and induced mutagenesis)
Question 102: Does the information before the Panel support the view that the biotech products
at issue in this dispute (including products subject to the member State safeguard measures)
give rise to the same types of potential risks to human, plant or animal health or to the
environment as novel non-biotech products, such as plant products produced by selective
breeding, cross-breeding and induced mutagenesis? If so, for any biotech product at issue in
this dispute are there significant differences, from a scientific perspective, in the nature or
magnitude of any potential risks from these products compared to comparable novel non-
biotech products taking into account:
• the specific genetic modification introduced and the resulting product;
• the intended use of each product (direct human or animal consumption, further
processing for consumption, planting or other use);
• any potential risks that may arise from the combination or successive use of
biotech products or comparable novel non-biotech products.
Please explain with reference to specific products at issue in this dispute.
Dr. Nutti
1149. Answer 102: It is important to point out that usually the novel non-biotech products, such as
plant products produced by selective breeding, cross-breeding and induced mutagenesis have not been
subjected to any extensive safety assessment as the plants derived from the Recombinant-DNA
technology have been. I am of the opinion that both novel non-biotech products and biotech products
should be assessed in a comprehensive, scientific, step by step , case by case bases, so the same safety
assessment principles should be applied in all cases.
Dr. Andow
1150. Comparable non-biotech products from plant bree ding: Regarding the differences between
conventional breeding and so-called "molecular breed ing," none of the parties fully represent the
comparison accurately. Breeding involves two im portant processes: (1) finding and introducing
usable genetic variation into the breeding population and (2) improving and selecting desired varieties
from the breeding population. The methods used will depend on the crop, with major differences for

clonal species, such as potato and banana versus sexual species, such as oilseed rape, maize and
soybean. Among the sexual species, there are ma jor differences in breeding methods between
outcrossing species, such as maize and oilseed ra pe, and inbreeding species, such as soybean, and
wheat.
1151. Transgenesis is a process to introduce genetic variation into the breeding population.
Transgenesis is not a process for finding genetic variation or for selecting and improving varieties.
Thus, there is no such thing as molecular breeding. Transgenesis is merely a part of the breeding
process.
1152. It would seem important then to determine a comparable conventional counterpart to
transgenesis for introducing genetic variation into the breeding population. A comparable
conventional process would be one typically used in breeding programs rather than one rarely used. If
possible a comparable conventional process would introduce genetic variation during a similar or
analogous step in the breeding process. The breed ing process can eliminate considerable genetic
variation, so variation introduced very early in the breeding process, will be screened and selected
multiple times by multiple breeding programs prior to use, while va riation introduced much later in
the process will receive fewer screenings and possibly no additional selection.
1153. Several other methods exist to introduce genetic variation into the breeding population.
These include wide crossing, crossing or using una dapted material, mutagenesis, and recombining
diverse adapted material. Wide crossing involves cr osses with different species. Unadapted material
are plants of the same species (or subspecies) th at have not previously been used in breeding
programs. Adapted materials are those that have already been used to produce modern conventional
varieties. There is a range of "adaptedness" ev en within these materials, with some breeding
programs relying primarily on the most recently us ed popular varieties and other programs reaching
further back in time for older varieties as the source of genetic variation.
1154. Crossing methods are difficult to use routin ely for clonal and inbreeding species. For
example, in potato, genetic varia tion is conventionally introduced by growing plants to flowering and
crossing material, or via deliberate or spontaneous mutation during cell culture. The crossing
methods take a longer time to produce usable variet ies. For example some characters have not been
incorporated in usable varieties despite over 30 y ears of work. Cell culture is essential for producing
new potato seed stock, so it is a routine way to introduce genetic variation and transgenesis introduces
genetic variation in cell culture. Thus for transgenic potatoes, the most relevant conventional
comparison is deliberate or spontaneous mutation in cell culture.
1155. For inbreeding species, such as soybean and dry bean, variation can be introduced by hand
pollinations (usually involving ad apted material), or by using new collections because inbreeding
lines will breed true to type. In some cases it is possible to create situations where the inbreeding
plant will outcross at higher rates. In this way, it is possible to introduce variation using crossing
methods. However, because the crossing methods typically are difficult, most programs do not use
them very much. All programs rely on evaluating co llections (sometimes an old collection in a new
environment) for new, usable genetic material. For these species, transgenesis is a new and powerful
method for introducing genetic variation directly into the adapted modern varieties, thereby avoiding
the expense of crossing and the arbitrariness of collections. Thus, it is not clear that there is a
comparable conventional method.
1156. For outcrossing species, such as maize and oil seed rape, there are many means to introduce
genetic variation. For modern maize breeding, however, wide crosses and crossing with unadapted
materials are not used in the vast majority of br eeding programs in the world. Many public sector

programs devote some resources to these, but they are viewed as long term breeding efforts that may
produce improved populations over decadal periods. These improved populations may or may not be
picked up by other breeding programs for introducti on into their unimproved breeding populations.
By far the most common method for introducing new genetic variation into a maize population for
breeding is recombination of adapted material. However, dwarfing even this method is
"backcrossing," which is a method for introducing a specific genetic trait into an adapted inbred line.
Both conventional and transgenic traits are introdu ced into adapted inbred lines via backcrossing.
However, for maize, recombination of adapted mate rial is probably the most relevant conventional
comparison to transgenesis.
Types of risks:
1157. As found by all scientific panels addressing this issue, there are no differences in the types or
kinds of risks posed by biotech crops compared wi th their non-biotech counterparts. The kinds of
risks include toxicity to humans and animals, allerg enicity, nutrition, potential for producing disease,
gene flow risks, non-target and biodiversity risks, and resistance risks.
Nature or magnitude of risks:
1158. Within these kinds of risk, there are new risks of biotech plants.
1159. The EC is correct in pointing out that both biolistic and Agrobacterium-mediated
transformation typically results in multiple transgene loci, typically with complex structure. 136 Unlike
bacterial transformation, where what is intended to be inserted typically is inserted just as intended,
this is not typical using the present plant transformation methods. One of the main concerns arising
from this are new open reading frames (ORFs). An ORF is a DNA sequence that could theoretically
produce a protein. A new ORF could theoretically produce a new protein, which could cause or
influence a new risk.
1160. The EC and the US is correct to identify in sertional mutagenesis as another outcome of
transgenesis that is new to breeding, and could cause or influence a new risk.
1161. Canada is incorrect to claim that transgenesis allows more precise control than selective
breeding. First both transgenic and conventional varieties undergo some level of selective breeding,
so the comparison is inappropriate. More importa ntly, transgenesis allows control over the intended
result (and in this sense, there is no dispute that transgenesis allows more precise control), but the
actual result is often different.
1162. The US, Canada and Argentina are correct to say that translocations and other genomic
disruptions can occur in conventional breeding. However, these genomic disruptions are normally
rare in conventional breeding. Moreover, their frequency of occurrence is much higher when genetic
variation is introduced using wide crossing than th e more typically used recombination of adapted
materials.
1163. Thus, the magnitude of ORFs and insertional mutagenesis introduced by transgenesis is
higher than that introduced by recombination of ad apted materials. It also may be higher than
deliberate or spontaneous mutation in cell culture, although this is less certain.

136 Please refer to additional detail in my response to question 9.

1164. There is considerable research being conducted to understand and control the transgene
insertion process, and it is likely that over the ne xt decade technical improvements will alter these
concerns.
1165. Some of the ORFs and extraneous transgene loci can be eliminated by independent
assortment for outbreeding plants, such as maize. This is less possible for inbreeding plants and
clonal plants.
1166. Nearly all risks associated with novel toxins (e.g., all Bt crops) introduced into crop plants are
new risks. While there are comparable risk asse ssment models for assessing these risks (toxins in
plants), and the risks of the particular toxins may have been investigated outside the plant, the fact that
the plant is used as the delivery vector for the t oxin and the precise expression patterns in the plant
mean that new species are exposed in new ways. These are new risks.
1167. Other risks of a new nature have been iden tified in some of my responses to the other
questions.
1168. The intended use of the product does not affect the nature of the risk, but it does affect the
rusk quantitatively. This is an effect of scale. For example, risks associated with releases for
processing are smaller than the same risks for the sa me product for planting and other use. It is not
clear how much smaller, but in some cases (biodive rsity risks of GMHT crops), it could be nil for
processing uses while it at the same time is was substantial for cultivation uses.
1169. Risks may change quantitatively from the comb ination of transgenes, because it will not be
possible to maintain seed supplies of all possible combinations of traits. Suppose a crop, such as
cotton, was available mainly with both a GMHT gene and at Bt gene. Suppose the insect pest is a
sporadic one on cotton, but a major one on maize. Farmers would have less incentive to use the Bt
gene, but may have a strong desire to use the GMHT gene. This would lead to an over use of the Bt
gene in cotton, increasing resistance risk both in cotton and maize.
Dr. Snow
1170. In my opinion, and only in reference to environmental risks, the biotech products in this
dispute could be considered as different from non-biotech products in the some cases, such as:
• Pesticide-producing crop plants –
– Non-biotech maize does not produce proteins that are toxic to insects,
including pollen that can kill lepidop terans (butterfly and moth lavae),
whereas Bt maize does (especially Bt-176, see NRC 2002). Here, one
concern is that drifting maize polle n could land on the food plants of
susceptible, non-target insects and kill them . Although this risk has largely
been discounted as the result of several i nvestigations in the USA, it is still a
question that scientific advisory panels must consider for all Bt crops. Other
non-target effects of Bt maize also require evaluation, including possible
unwanted effects on soil invertebrates, pollinators, ladybird beetles, and other
beneficial insects.

• Herbicide-tolerant crop plants with feral ( volunteer) populations or wild relatives that
can hybridise with the crop:
– See answers to Question 6 and others. Glyphosate-resistance has not been
introduced into crops using non-trangenic methods, so this particular trait is

linked to novel transgenic products . Because glyphosate is used so
extensively, weedy Brassica rapa populations that acquire the transgene for
glyphosate resistance might become more common in and around farmers'
fields. In the future, this process could occur many types of crops/weed
systems where weedy relatives of crops (e.g., weedy rice, chicory, carrot,
sunflower, sorghum, sugar beet, turf grass, etc.) are controlled using
glyphosate. Also, the herbicide glyp hosate provides certain environmental
benefits (such as facilitating no-till ag riculture and replacing more toxic
herbicides) that could be lost if there is extensive gene flow to volunteers and
wild relatives, and if over-use of the herbicide selects for other glyphosate-
resistant weeds. These problems are possible with all types of herbicide-
tolerant crops, unique features of glyphosate make it a special case.

Question 103: Does the information before the Pane l support the view that any of the biotech
products at issue in this dispute poses a substa ntially greater risk as regards the direct or
indirect consequences of unintentional "contamination" of other plant varieties than a
comparable novel non-biotech products, such as one of the 2300 different crop varieties that
have been developed using induced mutagenesis?
137 If so, what means of risk mitigation might
be envisaged?
Dr. Nutti
1171. Answer 103: My understanding is that plants that were developed us ing induced mutagenesis
should be subjected to the same risk assessment used for plants produced from the Recombinant-DNA
technology. In my opinion, crop varieties that have been developed using induced mutagenesis can
pose the same risk to human or animal health and, up to now, they have not been properly assessed. I
believe that guidelines should be developed for th e safety assessment of these plants, and that the
FAO/WHO Codex Guideline for the Conduct of F ood Safety Assessment of Foods Derived from
Recombinant-DNA Plants could be used as baseline.
Dr. Andow
Risk and "contamination":
1172. This will depend on the scale of release and the nature of the adverse effect.
1173. Risk is a combination of exposure and an adverse effect. Exposure will be determined by the
properties of the crop plant and the quantity of each planted (scale of release). There is little evidence
to suggest that ceteris parabis, gene flow will be greater from a transgenic variety than a conventional
one.138
1174. The definition of an adverse effect entails speci fication of who or what will be affected and
how is this effect considered adverse. Thus an organic farmer may find contamination adverse
because it removes his or her product from the organic food stream. This could be considered adverse
because of economic loss and lose of quality of life or livelihood.

137 FAO/IAEA (Food and Agriculture Organization of the United Nations/International Atomic Energy
Agency). 2001. FAO/IAEA Mutant Varieties Database. Available online at http://www-infocris.iaea.org/MVD/.
138 Some transgenic plants may have lower levels of gene flow, such as the male-sterile oilseed rape
varieties.

1175. There has been little discussion and less agreement over the nature of the adverse effects of
contamination. For example, there can be ec onomic and livelihood harms associated with the
perception of adverse effects. It is not clear to me that there is agreement that these kinds of
perceived risks should be considered.
1176. For the example in paragraph 1174, the risk associated with biotech crops is substantially
greater than the risk asso ciated with any of the conventionally pr oduced varieties. However, this is
largely due to the definition of the harm.
1177. Although there is little data to support this hypoth esis, I would suggest that the probability of
cross-contamination rises slowly with spatial scale for very small scale production, but once it reaches
a large-size threshold, the probability rises much fast er. In general, "contamination" will be a spread
process for which there is no advantage of the transg ene in the invaded/ contaminated habitat, but
there is an advantage in the habitat of origin. In these cases, spread scales with the length of the
boundary between the two habitats (not the area of either) and linearly with the square of time. 139
However, at very small spatial scales, it is likely th at managers will be able to take greater care in
limiting accidents, resulting in lower contamination rates.
Dr. Snow
1178. The question of whether any of the biotech pr oducts in this dispute poses a substantially
greater risk due to "contamination" than non-biot ech products depends on what types of risks each
party is trying to avoid (see Part I). If one considers that standard th resholds have been established to
allow the "coexistence" of biotech and non-biotech crops in Europe, then by definition contamination
is an issue for all biotech products. Very low le vels of contamination can be detected using DNA
markers, with the result that contamination, also known as "adventitious presence," has been detected
routinely in non-biotech products. Labeling a nd traceability have not been required for novel non-
biotech products such as those that were develope d using mutagenesis, nor would any high-resolution
identifying markers be available for these types of crops.
1179. Another way to answer this question is to focu s on the characteristics of biotech crops – their
phenotypes – rather than the mere pr esence of transgenes. This is more appropriate if the goal is to
avoid direct or indirect harms to human, plant or animal health, or the environment. The present
dispute involves only a few crop species with a limited number of transgenic traits, and the question is
whether the dispersal of these traits to other plants, by means of pollen and seed movements, could
pose substantially greater risks as compared to non-GM traits. See Part I and answers to Question 6
and others above for examples of risks that are gr eater in certain GM crops as compared to their non-
GM predecessors.
Dr. Squire
1180. A certain level of impurity – through one cr op type growing within or giving pollen to
another type – seems to have been generally accepted for many years. Such impurity varies greatly
with crop type: it is common in oilseed rape, and l ess common in beet and maize, for example. Where
the nature of the yield is different, e.g. for high erucic oilseed rape (HEAR), impurities are kept to
acceptably low values by using registered growers who are familiar with the crop and with the
requirements for separation, and many of whom farm in a restricted part of the country. There are
clear criteria for the acceptable presence of HEAR in non-HEAR oil. Outside specialist varieties such

139 This is much slower than for species invasions, which scale on the square root of the area occupied
and linearly with time.

as HEAR, impurities have largely gone unobserved, and in many instances where non-GM oilseed
rape volunteers emerge in a non-GM oilseed rape crop, the percentage impurity is unknown and is
ignored. The potential of a crop variety to convey an impurity is not the same for all varieties but
differs depending on factors such as the proportion of seeds that enter dormancy, the persistence of
seeds in the soil and the relative pollen 'strength' of potential donor and recipient fields (for example
caused by different proportions of male sterile pl ants). These factors may or may not differ between
GM and non-GM varieties but should not differ be cause one is GM and the other not. They would
differ in any case. Given present knowledge of the life cycle and reproductive behaviour of the crops,
there is no reason to suppose that biotech crops confer different degrees of impurity compared with
crops produced from, say, induced mutagenesis.
Question 104: From a scientific perspective, is there a significant difference in risks to human,
animal or plant health or the environment ar ising from the use of a bacterial antibiotic
resistance marker gene, or part thereof, in any biotech product at issue in this dispute (eg,
Monsanto Bt cotton (531), Monsanto Ro undup Ready cotton (RRC1445), Amylogene starch
potato) compared to novel non-biotech products, such as comparable plant products produced
by selective breeding, cross-breeding and induced mutagenesis?
Dr. Nutti
1181. Answer 104: As I pointed out before, the use of antibiotic resistant marker genes has been
recognized as a safe tool and its employment should be evaluated on a case by case basis. The same
rule should be applied to novel non-biotech produc ts, such as comparable plant products produced by
selective breeding, cross-breeding and induced mu tagenesis. I mean, these products should be
evaluated on a case by case basis, so the same safety assessment principles should be applied in all
cases.
Question 105: For those biotech products at issue in this dispute for which no significantly
different nature or level of risk has been identified, is there a scientific or technical rationale for
monitoring the occurrence of pote ntial adverse effects, or of unintentional effects, arising from
the consumption or use of these products compared to novel non-biotech products, such as plant
products produced by selective breeding, cross-b reeding and induced mutagenesis? If so, would
such monitoring relate to the specific genes or tr aits introduced into a biotech product, and how
would this compare with the monitoring of induced changes in novel non-biotech products?
Dr. Nutti
1182. Answer 105: I am of the opinion that the monitoring of occurrence of potential adverse
effects or of unintended effects should be carried out based on scientific parameters in both cases, that
is, for those biotech products at issue in this dispute, for which no significantly different nature or
level of risk has been identified, and for novel non- biotech products, such as plant products produced
by selective breeding, cross-breeding and induced mutagenesis. In both cases, I am considering that
the products were assessed as safe.
1183. It is important to point out that usually the novel non-biotech products, such as plant products
produced by selective breeding, cross-breeding and i nduced mutagenesis have not been subjected to
any extensive safety assessment as the plants derived from the Recombinant-DNA technology have
been.
1184. I also think that the methodologies which will be used for this monitoring need to be
evaluated very carefully since it will be very difficult to monitor any effect that we are not aware of.

Therefore, I think this is one good point to be in troduced in the agenda of the Codex Alimentarius
Task Force on Foods Derived from Biotechnology, which will start working in September 2005.
Dr. Andow
Rationale for monitoring:
1185. There are some biotech products at issue in th is dispute for which one committee or another
has determined that were no significant risks. I have taken issue with some of these determinations in
my previous responses. For all of the products at issue in this dispute, there are some member
countries that disagree with the assessment of the relevant SCP (for example) that there are no
significant identified risks. This disagreement is sometimes related to different standards of
acceptable risk being applied by the disagreeing parties.
1186. This disagreement provides the only rationale for monitoring potential adverse effects. If all
parties agreed that there were no significant iden tified risks, there would no need to consider
monitoring for potential adverse effects.
1187. Unanticipated effects could arise from a transg ene by any of the following mechanisms: new
ORFs, insertional mutagenesis, post-transcriptional or post-translational processing, other pleiotropy
or epistasis, and gene-by-environment interaction. New ORFs and insertional mutagenesis can be
addressed through molecular characterization of all transgene loci. Post-transcriptional or post-
translational processing can be addressed to the most part by molecular and biochemical analysis.
Other pleiotropy or epistasis and gene-by-envi ronment interaction cannot be addressed without
extensive planting in the field.
1188. Under present technologies, transgenes can generate new ORFs and insertional mutagenesis.
Post-transcriptional and post-translational processing may relate to the structure of the transgene and
the nature of the gene product. Products novel to the plant probably require closer assessment. A
gene product native to the recipient plant may be less subject to processing, except if it is expressed in
new tissues. Novel products probably require cl oser assessment than native products for other
pleiotropy or epistasis and gene-by-environment interaction.
1189. Monitoring can be used to look for unanticipated effects.
1190. A less thorough molecular and biochemical ch aracterization of the transgene locus and
transgene products provides increased potential for unanticipated effects.
1191. Monitoring may be partially substitutable for molecular and biochemical characterization.
1192. Monitoring may also substitute for identifying all possible effects, by covering various
categories of unanticipated effects. If a thor ough molecular analysis is conducted, it would seem
important to ensure that monitoring for the remaining unanticipated effects is not too expensive.
Comparison with non-biotech varieties:
1193. For monitoring related to a specific potential adver se effect, this would have to relate to the
specific genes/ traits in the transgenic crop. For monitoring for unanticipated effects, a more general
approach is needed.

1194. First, I should make clear that some conventiona l non-transgenic plants have risks, some of
which may justify monitoring (and management).
1195. For potential adverse effects, the reasoning I have used here implies that the potential adverse
effect is not a potential adverse effect from a conventional non-transg enic plant. Otherwise there
would be no disagreement as I have posited in paragraph 1185.
1196. For unanticipated effects, there is a quantitative difference in some of the concerns related to
the process of transgenesis (paragraph 1162), and differences related to the novelty of the transgene
product (paragraph 1188). Native biotech products may or ma y not be different from conventional
breeding for unanticipated effects stemming from other pleiotropy or epistasis and gene-by-
environment interaction.
Dr. Snow
1197. My conclusion is that it does not seem logical to require monitoring if no risk has been
identified (see Question 99). Also, both transgenic methods and other methods used crop breeding
can result in unintended effects, and in my opini on there is no basis to assume that these unintended
effects would be substantially greater in transgenic plants.
Question 106: For those biotech products at is sue in this dispute for which an approval has
been sought for environmental release (notif ications submitted under Directives 90/220 or
2001/18), and for which no significantly different nature or level of risk has been identified, does
the information before the Panel provide any sci entific or technical rationale for requiring
specific agricultural management practices that differ from those for novel non-biotech
products, such as plant products produced by selective breeding, cro ss-breeding and induced
mutagenesis?
Dr. Andow
Rationale for management:
1198. There are some biotech products at issue in th is dispute for which one committee or another
has determined that were no significant risks. I have taken issue with some of these determinations in
my previous responses. For all of the products at issue in this dispute, there are some member
countries that disagree with the assessment of the relevant SCP (for example) that there are no
significant identified risks. This disagreement is sometimes related to different standards of
acceptable risk being applied by the disagreeing parties.
1199. This disagreement provides the only rationale for risk management. If all parties agreed that
there were no significant identified risks, there wo uld no need to consider monitoring for potential
adverse effects. If only some of the parties recognized the risk, then some kind of conditional risk
management could be justified. For example, one condition for the management measures could be
the country of use.
Dr. Squire
1200. Provided no significant level or type of risk has been detected, then no particular change in
practice above the highest recommended existing prac tice to ensure a high purity, should be needed.
As in Q.103, however, if thresholds are imposed fo r whatever reason, e.g. 0.9%, specifically for GM
varieties in non-GM varieties, then there will need to be different agricultural practices for those GM

varieties that leave volunteers or spread genes by pollen to neighbouring, sexually compatible crops.
These practices are well appreciated – reducing volunteers to a minimum by appropriate soil
cultivation after harvest, preventing volunteers from flowering in subsequent crops, leaving several
years before growing the same type of crops again so that volunteer seed decays by natural means,
leaving a separation strip between crops which is not harvested and beyond which geneflow is
reduced to very low values, and not planting crops having reduced self-pollen in the vicinity.
1201. Under a system of coexistence in which a threshold (GM in non-GM) was imposed, the
agricultural practice might well have to change in a crop such as oilseed rape to ensure that threshold
would be met. Longer intervals than normal between oilseed rape crops, some regional segregation of
GM and non-GM crop types and the dropping of vari etal associations (80% male sterile, 20% own
pollen) from general use would probably be necessary. Given present knowledge, these changes
would be in consequence of an imposed threshol d of GM in non-GM product, not of any inherent
food-risk in the GM product itself.
Foods produced with biotech processing aids 140, including yeasts, bacteria or enzymes that have
been modified using recombinant DNA technology
Question 107: Does the information before the Panel support the view that the biotech products
at issue in this dispute (including products subject to the member State safeguard measures)
give rise to the same types of potential risks to human, plant or animal health or to the
environment as foods produced using biotech processing aids, including yeasts, bacteria or
enzymes that have been modified using recombin ant DNA technology? If so, for any biotech
product at issue in this dispute are there signif icant differences, from a scientific perspective, in
the nature or magnitude of any potential risks from the products at issue in this dispute
compared to foods produced with biotech proces sing aids, taking into account the intended use
of each product (direct human or animal cons umption, further processing for consumption,
release into the environment, any other use).
Dr. Nutti
1202. Answer 107: According to the information before the Panel, and the existing bibliography in
this field, my view is that the safety of the biotech products at issue in this dispute (including products
subject to the member State safeguard measur es) should be assessed according to the FAO/WHO
Codex Guideline for the Conduct of Food Safety Assessment of Foods Derived from Recombinant-
DNA Plants, in order to evaluate the potential risks to human, plant or animal health.
1203. For foods produced using biotech processing aids , including yeasts, bacteria or enzymes that
have been modified using recombinant DNA technology, it is important to point out that the
microorganisms which produces the processing aid was modified and NOT the processing aid itself.
Therefore, I am of the opinion that the safety of the new microorganisms and of the product thereof
should be assessed according to the FAO/WHO Codex Guideline for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Microorganisms.

140 "Processing aid" is defined by the Codex Alimentarius as "any substance or material, not including
apparatus or utensils, and not consumed as a food ingredient by itself, intentionally used in the processing of
raw materials, foods or its ingredients, to fulfil a cer tain technological purpose during treatment or processing
and which may result in the non-intentional but unavoidable presence of residues or derivatives in the final
product".

1204. It is important to recognize that in both cas es the safety assessment should be carried out in
order to evaluate the existence of any potential risks. If the intended use of the two different
categories of products (foods and processing aids) is taken into account, the first point to be
considered is that processing aids are used in ve ry small amounts in food products, so the direct
human or animal consumption of processing aids obtained through the recombinant DNA-technology
will be much lower than foods derived from Recombinant-DNA plants. As far as further processing
for consumption is concerned, most of the foods that are consumed nowadays is subjected to
processing, so this point cannot be considered as a difference among the two product categories.
Dr. Andow
Kinds of risk:
1205. In general, foods produced with biotech processing aids are not expected to be able to self-
reproduce in the environment. Thus the kinds of risk associated with these foods do no include any of
the environmental risks associated with transgenic plants.
1206. Foods produced with biotech processing aids can ha ve some non-target effects if the food is
available in the environment. These effects are more similar to the effects of chemicals rather than
the effects of living biological organisms.
Nature and magnitude of risk:
1207. I will not address human or animal health hazards. The following comment, however, has
some implications for human and animal health hazards.
1208. As I have argued in question 9 and 102, in tran sgenic plants, unanticipated effects could arise
from a transgene by any of the following mechan isms: new ORFs, insertional mutagenesis, post-
transcriptional or post-translational processing, other pleiotropy or epistasis, and gene-by-environment
interaction. For each of the potential sources of una nticipated effects, the likelihood of it occurring in
a transgenic bacterium or yeast (that is typically u sed as a food processing aids) is much lower than in
a transgenic plant. For transgenic bacteria and fungi (especially yeast), transgene insertion is more
predictable and the probability of new ORFs is much lower. Transgenic bacteria and yeast are much
easier to screen and eliminate insertional mutations. Moreover, there is considerably less post-
transcriptional and post-translational processing of gene products in bacteria and yeast than in plants.
Finally the genome size is smaller in bacteria and y east and they are used in a much more restricted
set of environments than plants, so risks associated with other pleiotropy and epistasis and gene by
environment interaction are less.
Question 108: Is there a significant difference in risks to human, animal or plant health or the
environment arising from the use of a bacteri al antibiotic resistance marker gene, or part
thereof, in any biotech product at issue in this dispute (eg, Monsanto Bt cotton (531), Monsanto
Roundup Ready cotton (RRC1445), Amylogene starch potato) compared to foods produced
with biotech processing aids, including yeasts, bacteria or enzymes that have been modified
using recombinant DNA technology?
Dr. Nutti
1209. Answer 108: As I pointed out before, the use of antibiotic resistance marker genes has been
recognized as a tool that can be safely used, and that its use should be evaluated on a case by case
basis. The same applies to processing aids, that is, the processing aids derived from Recombinant-

DNA microorganisms should be evaluated on a case by case basis. Therefore, the same safety
assessment principle should be applied in both cases.
Question 109: For those biotech products at issue in this dispute for which no significantly
different nature or level of risk has been id entified, does the information before the Panel
provide a scientific or technical rationale for monitoring the occurrence of potential adverse
effects, or of unintentional effects, arising from the consumption or use of these products
compared to foods produced with biotech processing aids? Does the information before the
Panel provide a scientific or technical rationale for mitigating any potential risks arising from
the biotech products at issue in this dispute in a manner differently than products produced
with biotech processing aids?
Dr. Nutti
1210. Answer 109: For those biotech products at issue in this dispute for which no significantly
different nature or level of risk has been iden tified and for foods produced with biotech processing
aids, where the processing aid and the Recombinant-DNA microorganism have been assessed as safe,
I am of the opinion that the monitoring of occu rrence of potential adverse effects or of unintended
effects, in both cases, should be conducted based on scientific parameters.
1211. I also think that the methodologies which will be used for this monitoring need to be
evaluated very carefully since it will be very difficult to monitor any effect that we are not aware of.
Therefore, I think this is one good point to be in troduced in the agenda of the Codex Alimentarius
Task Force on Foods Derived from Biotechnology, which will start working in September 2005.
Dr. Andow
Monitoring and Mitigation:
1212. There are some biotech products at issue in th is dispute for which one committee or another
has determined that were no significant risks. I have taken issue with some of these determinations in
my previous responses. For all of the products at issue in this dispute, there are some member
countries that disagree with the assessment of the relevant SCP (for example) that there are no
significant identified risks. This disagreement is sometimes related to different standards of
acceptable risk being applied by the disagreeing parties.
1213. This disagreement provides the only rationale fo r monitoring or mitigating potential adverse
effects. If all parties agreed that there were no significant identified risks, there would no need to
consider monitoring for potential adverse effects.
1214. Unanticipated effects could arise from a transg ene by any of the following mechanisms: new
ORFs, insertional mutagenesis, post-transcriptional or post-translational processing, other pleiotropy
or epistasis, and gene-by-environment interaction. New ORFs and insertional mutagenesis can be
addressed through molecular characterization of all transgene loci. Post-transcriptional or post-
translational processing can be addressed to the most part by molecular and biochemical analysis.
Other pleiotropy or epistasis and gene-by-envi ronment interaction cannot be addressed without
extensive planting in the field.
1215. All of these possible sources unanticipated effect s are more likely to occur for plant biotech
products than for foods produced with biotech processing aids. This provides a rationale for
monitoring, but no rationale for mitigation, as there are no concrete effects that can be mitigated.

Question 110: On the basis of the information be fore the Panel, is there any scientific evidence
to support the hypothesis that animal feed made from biotechnology plants alters the
composition of the food derived from animals consuming the feed?
(a) If so, what is the likelihood that this event could lead to adverse effects on
human or animal health? (see, inter alia, exhibit US-144)
(b) What risk management options are available to mitigate any resulting risks and
what is their efficacy?
Dr. Nutti
1216. Answer 110: I am not aware of any scientific evidence that supports the hypothesis that
animal feed made from biotechnology plants alters the composition of the food derived from animals
consuming the feed. One of the studies provided at US-144 showed no difference in milk produced
from lactating cows fed rations with genetically modified whole cottonseed to milk produced from
cows fed non genetically modified but genetically similar cottonseed. The study showed that no milk
samples were positive for tr ansgenic or plant DNA fragments at the limits of detection for the assays
following detailed data evaluation criteria. The DMI, milk yield, milk composition, body weight and
body composition score did not differ among treatment s. The study showed that cottonseed from
genetically modified varieties used in these studi es yielded similar performance in lactating dairy
cows when compared to non transgenic control and reference cottonseed.
1217. The second study used dehulled soybean meal prepared from genetically modified, herbicide
tolerant soybeans and near isogenic conventional soybeans in an experiment with growing-finishing
pigs. The study concluded that rate and efficien cy in weight gain, scanned backfat ,longissimus area
and calculated carcass lean percentage were not diffe rent for pigs fed diets containing conventional or
Gm soybean meal. Composition of the meat and sens ory scores were not influenced by diet. The
results indicated that Roundup Ready soybean meal is essentially equivalent in composition and
nutritional value to conventional soybean meal for growing-finishing pigs.
1218. These are not the only studies that had been ca rried out for the comparison of composition of
the food derived from animals consuming GM grains, pl ants or meals. Dr. Marjorie Faust, from Iowa
University has conducted this type of studies since 1997, with different crops and different animals,
and up to now, the results showed no difference in the composition of the products or in the animal
performance.141

141 References:
Faust, M., and L. Miller.1997 Study finds no Bt in milk. ISU International Crop Management
Nwesletter ICC-478. Fall Special Livestock Edition. pp 6-7. Iowa State University Extension, Ames.
Faust, M., and S.M. Spangler. 2000. Nutritive valu e of silages from MON810Bt and non- Bt near
isogenic corn hybrids. J. Dairy. Sci.83:1184.
Hammond et al.1996 The feeding value of soybeans fed to rats, chickens, catfish and dairy cattle is not
altered by genetic incorporation of glyfosate tolerance. J.Nutr.126:717-727.
Padgett at all 1995. Development, identification and characterization of a glyphosate –tolerant soybean
line. Crop Sci.35:1451-1461.
ILSI ( 2004) "Nutritional and Safety Assessments of Foods and Feeds Nutritionally Improved through
Biotechnology". Comprehensive Reviews in Food Science and Food Safety at
http://www.ift.org/pdfs/crfsfs/crfsfsv3n2p0035-0104ms20040106.pdf.

Question 111: Please provide an assessment of th e US statements regarding the evaluation of
the safety of biotech products in paragra phs 128-133 of the US Supplementary Rebuttal
Submission, and in particular of the statement in the last sentence of paragraph 133 that:
"...where all of the data consistently provide no indication of adverse effects, and there is no
specific indication that the data submitted are in adequate, there is generally no reason to expect
that any remaining risk as gone undetected, and that further studies are warranted". What
relevance does the foreseen end product use(s) have in the context of the evaluation of the safety
of that product?
Dr. Nutti
1219. Answer 111: As I had pointed out in several answers, and based on the Codex Alimentarius
Guidelines for the Conduct of Food Safety Asse ssment of Foods Derived from Recombinant-DNA
Plants (CAC/GL 45-2003), paragraphs 34 to 43, A ssessment of Possible Toxicity, we have at
paragraph 37 that the use of a ppropriate conventional toxicol ogy or other studies on the new
substance may be necessary if, taking into account its function and exposure, doubts about the safety
of the new substance remain. My understanding is that if the toxicological assessment has
presented safety studies on protein digestion in mammalian gastric and intestinal systems, acute
gavage studies in mice, homology to known toxins and allergens, and exposure to human diet
that has no indication of adverse effects, and the comparison of the GM and its conventional
counterpart has been properly conducted and showed equivalence, there is generally no reason
to expect any remaining risk.
1220. The relevance of the end product use(s) is consid ered in the safety assessment, if we consider
that we have the comparison not only between the GM grain or plant and its conventional counterpart,
but we also have to compare the composition of prod ucts thereof, for instance in the case of soybeans
the comparison has considered different processed pr oducts as: soy meal, defatted soy meal, toasted
flour, non toasted flour, refined oil, isolated protein and extruded soy protein.
Dr. Snow
1221. In general, the scientific arguments in para graphs 128-133 are valid, but the US does not
acknowledge possible inherent shortc omings of small-scale, short-duration field tests. The US
correctly states that: "Such tests are designed prim arily to examine whether the bioengineered plant
exhibits the intended characteristics." However, these tests rarely address environmental risks such as
the potential consequences of interbreeding with wild or weedy relatives, as the US claims. In
paragraph 131, the US states that "effects on ot her organisms in the ecosystem" are evaluated,
including "indirect impacts, such as those on fa rming production methods". However, the methods
for making these evaluations are not explained or cited.
1222. Here, I think it is important to point out that these same questions are central to many of the
concerns that several EC Member States have raised . These are the types of questions that sometimes
suggest the need for further research, which takes time, resources, and multidisciplinary
collaborations. The last sentence of paragraph 133 does not acknowledge that important scientific
information about environmental consequences could be lacking in specific cases, in which case
further scientific studies would be warranted.
1223. The foreseen end product uses of the product are relevant to safety evaluations – this is a
standard assumption in all risk assessments, so I am not sure what the point of this question is.

Question 112: Please provide an assessment of the statements regarding chronic toxicity testing
in paragraph 43 of Canada's Third Submission and paragraphs 134-138 of the US
Supplementary Rebuttal Submission. What relevan ce does the foreseen end product use(s)
have in the context of the evaluation of the toxicity of that product?
Dr. Nutti
1224. Answer 112: As I had pointed out in several answers, and based on the Codex Alimentarius
Guidelines for the Conduct of Food Safety Asse ssment of Foods Derived from Recombinant-DNA
Plants (CAC/GL 45-2003), paragraphs 34 to 43, A ssessment of Possible Toxicity, we have at
paragraph 37 that the use of a ppropriate conventional toxicol ogy or other studies on the new
substance may be necessary if, taking into account its function and exposure, doubts about the safety
of the new substance remain. My understanding is that if the toxicological assessment has conducted
acute gavage studies in mice, homology to known toxins and allergens with no indication of adverse
effects, chronic toxicity tests shall not be required.
Question 113: Please provide an assessment of th e statements regarding the purpose and use of
whole food studies in paragraphs 142-144 of th e US Supplemental Rebuttal Submission. What
relevance does the foreseen end product use(s) have in the context of using whole food studies in
the evaluation of the safety of that product?
Dr. Nutti
1225. Answer 113: Based on the Codex Alimentarius Guidelines for the Conduct of Food Safety
Assessment of Foods Derived from Recombinant-DNA Plants (CAC/GL 45-2003), paragraphs 53, my
understanding is that additional animal feeding studi es may be warranted for GM foods if changes in
the biovailability of the nutrients are expected or if the composition of the GM food is not comparable
to conventional food.. My understanding is that if the comparison of the composition of the GM
product and its conventional counterpart has shown to be substantially equivalent, the 90 –day oral
study and at least one more feeding study ( usually 48 day broiler chicken ) has provided information
on the consumption of the whole food, the request for multiple whole food studies in different species
does not have scientific support.
Question 114: Please provide an assessment of the statements regarding small random DNA
insertions and molecular characterization in paragraph 152 of the US Supplemental Rebuttal
Submission.
No Answers.