Erläuterungen zu den Dateien des Monsanto MON810 Monitoring

Transcription

Erläuterungen zu den Dateien des Monsanto MON810 Monitoring
und Implementierungsplan
1. Erste Stellungnahme der Firma Monsanto mit Adressierung der im BVLBescheid gelisteten Punkte, die im Monitoring für MON810 berücksichtigt
werden sollen. Diese Stellungnahme enthält auch den von Monsanto auf EU
Ebene eingereichten Monitoringplan (‚EU-Monitoringplan’) für den Antrag
auf Erneuerung der Zulassung von MON810 Mais nach 1829/2003/EC in
Verbindung mit der RL 2001/18/EC.
2. a-c: Zweite Stellungnahme der Firma Monsanto, wie der EU-Monitoringplan
in Deutschland implementiert werden soll, mit dem Annex I (Angaben zum
Auswahlverfahren von bestehenden Umweltbeobachtungsprogrammen in
Deutschland) und dem Annex II (Liste von
Umweltbeobachtungsprogrammen in Deutschland, die Monsanto auf ihre
Eignung geprüft hat)
3. Dritte Stellungnahme der Firma Monsanto mit Angaben, wie Berichte von
ausgewählten Beobachtungsprogrammen ausgewertet werden.
4. Vierte Stellungnahme der Firma Monsanto, in dem ein zusätzliches
Bodenbeobachtungsprogramm in die Liste des Implementierungsplanes
aufgenommen wird mit der Erläuterung, wie geeignete Rohdaten von
Beobachtungsprogrammen generell analysiert werden sollen.
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Response to the German safeguard measure on
MON 810 maize
May 2007
INTRODUCTION
Maize is an important crop in the European Union (EU) and infestation by the
lepidopteran pest Ostrinia nubilalis (ECB - European corn borer) can result in
considerable crop damage and yield loss. In Germany, an estimated 500 000 ha
of maize experiences problems with ECB, in particular Bavaria and BadenWurtenberg (Brookes, 2007).
The use of conventional insecticides to protect against corn borers is not
practical, since chemical sprays cannot reach larvae that have bored into the
maize stalks. Crops that have been genetically modified to express a specific
Bacillus thuringiensis (Bt) protein toxic to corn borers offer an efficient
alternative to combat these pests.
Since its introduction in the USA in 1996, insect-protected Bt maize has proved
to be a successful management tool to control crop damage and yield losses due
to insect pests. This technology has been widely adopted and its use extended to
other countries. The Bt maize MON 810 has now been grown worldwide on a
cumulative area of approximately 80 million ha1.
In Europe, Bt maize was planted for the first time in 1998 in Spain. Bt maize
has been grown in Germany every year since then and is also grown
commercially in France, Portugal and the Czech Republic. In total, the area
planted to Bt maize in the EU was about 65 000 ha in 2006 (Brookes, 2007).
BACKGROUND
On April 27 2007, the German Federal Office of Consumer Protection and Food
Safety (BVL) temporarily suspended the authorization to distribute MON 810
maize seeds for commercial planting in Germany. The suspension is valid until
Monsanto, as authorisation holder, submits an appropriate monitoring plan for
MON 810 cultivation in Germany to the BVL. The Order invokes the safeguard
provisions of Directive 2001/18/EC, and as scientific justification, cites a
selection of scientific papers on the impacts of GM crops to non-target organisms
and risks for the soil.
MONITORING:
THE EU
CURRENT LEGAL REQUIREMENTS FOR
MON 810
CULTIVATION IN
Following the procedures required by EU Directive 90/220/EEC, MON 810
cultivation in the EU was authorized according to Commission Decision
1
http://www.monsanto.com/monsanto/content/investor/financial/reports/2006/Q42006Acreage.pdf
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MON 810 maize
May 2007
98/294/EC of 22 April 1998 and the subsequent consent issued by France2 in
accordance with this Commission decision.
Council Directive 90/220/EEC of 23 April 1990 on the deliberate release into the
environment of genetically modified organisms required applicants to submit
information on “monitoring, control, waste treatment and emergency response
plans” (Article 5 (2) (a)). This included methods for tracing the GMOs and for
monitoring their effects, specificity (to identify the GMOs, and to distinguish
them from the donor, recipient or, where appropriate, the parental organisms),
sensitivity and reliability of the monitoring techniques; techniques for detecting
transfer of the donated genetic material to other organisms and duration and
frequency of the monitoring.
Commission Decision 98/294/EC of 22 April 1998, authorizing MON 810
cultivation in the EU, establishes that: ”the notifier has defined a management
strategy in order to minimise the development of insect resistance and has offered
to inform the Commission and/or the Competent Authorities of Member States of
the results of monitoring of this aspect”. Following the recommendation of the
Decision 98/294/EC, Monsanto has complied with the requested monitoring
conditions and implemented an insect resistance management (IRM) plan since
the first placing on the market of MON 810 (in Spain in 2003).
Further requirements for mandatory post-market monitoring of newly
authorized GMO products were introduced with Directive 2001/18/EC of 12
March 2001. The European legal framework on GMO requires the notifier to
implement a monitoring plan for environmental effects confirming with Annex
VII of 2001/18/EC. Its objectives are to:
• Confirm through case-specific (CS) monitoring than any assumption
regarding the occurrence and impact of adverse effects of a GMO or its use
in the environmental risk assessment (e.r.a.) are correct.
• Identify through general surveillance (GS) the occurrence of adverse effect
of the GMO or its use on human health and the environment which were
not anticipated in the e.r.a.
To help the notifiers to implement these two aspects of the monitoring plan, the
GMO Panel of the European Food Safety Authority (EFSA) was mandated by the
European Commission to develop a guidance document (EFSA, 2006b) detailing
the requirements for both the CS and GS aspects of monitoring.
For products previously approved according to Directive 90/220/EEC, the
additional monitoring requirements of Directive 2001/18/EC are applicable upon
2
http://www.admi.net/jo/19980805/AGRP9801537A.html
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MON 810 maize
May 2007
renewal of the consent under Directive 2001/18/EC or Regulation (EC) No
1829/2003. However, in accordance with its commitment to product stewardship,
Monsanto proactively implemented a comprehensive monitoring plan in 2005.
This plan, which was subsequently updated and presented to support the
renewal of the notification for cultivation of MON 810 under Regulation (EC) No
1829/2003 in 2007, is in accordance with the monitoring requirements of
Directive 2001/18/EC and the EFSA guidance document.
STATE-OF-THE-ART MONITORING PLAN FOR MON 810
Environmental risk assessment (e.r.a.)
A complete environmental risk assessment was conducted according to Annex II
of Directive 2001/18/EC on the deliberate release of genetically modified
organisms in the environment. Nine potential direct or indirect, immediate or
delayed adverse environmental effects are laid out in Annex II of Directive
2001/18/EC, which could theoretically occur when a GM higher plant (GMHP) is
placed on the market in the European Union. The potential for MON 810 to
adversely impact the environment in the EU was evaluated according to the
recommended step-wise risk assessment process, including: a) identification of
potentially harmful characteristics of the GMHP; b) potential consequence of the
theoretical adverse effect (assuming fully realised); c) likelihood of the
theoretical adverse effect to occur for this product and a characterisation of the
actual hazard potential of the identified GMHP characteristic; d) estimation of
the risk and e) necessary risk mitigation measures, if applicable. MON 810 was
shown not to be different from conventional maize in its agronomic, phenotypic,
compositional, nutritional and safety characteristics, indicating that any
interactions of this maize with the biotic environment have not been changed
compared to conventional maize. No adverse environmental effects were
identified for MON 810 (see Appendix 1 for detailed e.r.a.).
Case specific monitoring
The objective of case-specific monitoring is to confirm that any assumptions
regarding the occurrence and impact of potential adverse effects of the GMO or
its use that have been identified in the e.r.a., are correct.
Since the conclusions of the e.r.a. (see Appendix 1) consistently show that the
placing on the market of MON 810 poses negligible risk to human and animal
health and the environment and since the conclusions of this e.r.a. are derived
from the results of scientific studies rather than assumptions, no case-specific
post-marketing monitoring actions, typically aimed at testing assumptions made
in this assessment, are considered necessary. The commercial cultivation of
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MON 810 maize
May 2007
MON 810 is, however, accompanied by stewardship programmes, including an
insect resistant management measures (provided in Appendix 2) with the
objective of reducing the potential for the target insects to develop resistance to
the Cry1Ab protein.
General Surveillance
The objective of general surveillance is to identify 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.
General surveillance is largely based on routine observation and implies the
collection, scientific evaluation and reporting of reliable evidence, in order to be
able to identify whether unanticipated, direct or indirect, immediate or delayed
adverse effects might have been caused by the placing on the market of a GM
plant in its receiving agricultural or non-agricultural environment. By nature,
the prediction of unanticipated effects does not lend itself to the formulation of
clear scientific hypotheses, and therefore it will need adapted scientific
methodology, as described in Section 11.4.3 of the GS plan that is detailed in
Appendix 3.
General surveillance is focused on the geographical regions within the EU where
the GM crop is grown, and is taking place in representative environments,
reflecting the range and distribution of farming practices and environments
exposed to GM plants and their cultivation.
Where there is scientifically valid evidence of a potential adverse effect (whether
direct or indirect), linked to the genetic modification, then further evaluation of
the consequence of that effect should be science-based and compared with
baseline information. Relevant baseline information will reflect prevalent
agricultural practice and the associated impact of these practices on the
environment. In many cases it may not be possible to establish a causal link
between a potential adverse effect and use of a particular GM crop
For general surveillance of MON 810, the party placing MON 810 on the market
uses several tools. An important tool is an annual farm questionnaire addressed
to a subset of farmers cultivating MON 810. This tool is complemented by
company stewardship programmes and a detailed analysis of the ongoing
scientific literature, and unsolicited reports. from various sources. Additionally,
information from other sources (,official websites and existing observation
networks) is incorporated, where appropriate.
The general surveillance performed in 2005 consisted of two main elements, firstly
a farmer questionnaire (considering the experimental questionnaire develop by the
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MON 810 maize
May 2007
German Federal Biological Research Centre for Agriculture and Forestry (BBA),
maize breeders and statisticians in Germany) that was designed to assess
observations in the areas where MON 810 has been cultivated and, secondly, an
assessment of the research work reported in peer reviewed publications in 2005
and 2006 that relate to MON 810 and its environmental safety.
1) Questionnaire
Farmers are the closest observers of the cultivation of the GM crops and
already collect information on the cultivation and management of their crops
at farm level. Therefore they can provide details on GM plant-based
parameters (referring to species/ecosystem biodiversity, soil functionality,
sustainable agriculture, or plant health) and on background and baseline
environmental data (e.g. soil parameters, climatic conditions, general crop
management data e.g. fertilizer use, crop protection measures, crop rotations
and previous crop history). Additionally farmers may provide empirical
assessments which can be useful within general surveillance to reveal
unanticipated deviations from what is common for the crop and cultivation
area in question, based on their historical knowledge and experience.
A questionnaire addressed to farmers cultivating MON 810 is a monitoring
tool that is specifically focused on the farm level. EFSA explicitly considers
questionnaires a useful method to collect first hand data on the performance
and impact of a GM plant and to compare the GM plant with conventional
plants (EFSA, 2006). The questionnaire approach has also proven its
applicability with other industries, e.g. the pharmaceutical industry.
Farmers have been asked for their observations and assessment in and
around MON 810 cultivated fields in comparison to a baseline, this being
their own historical local knowledge and experience. In 2005 this general
surveillance for MON 810 focused on the geographical regions within the EU
where MON 810 was grown commercially (Czech Republic, France, Portugal,
Spain). It was also performed in areas reflecting the range and distribution of
farming practices and environments of MON 810 cultivation. This allows for
cross-checking of information indicative of an unanticipated effect, and the
possibility to establish correlations either by comparing questionnaires
between regions, or associating answers to observations made by existing
networks, such as meteorological services (weather conditions) or extension
services (pest pressure).
In 2005, a subset of farmers in Spain (65) and almost all farmers in the Czech
Republic (five), France (38) and Portugal (24) were asked to fill in the
questionnaire.
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MON 810 maize
May 2007
It contained five sections:
1. Personal data
2. The farm and farm activities
3. Bt maize specific measures
4. Observations related to MON 810 maize in the field
5. Observations after harvest of MON 810 maize
Section 1 gave the coordinates of the farmer which are treated as confidential
information. Section 2 was designed to obtain information on the area of
arable land and the proportion of maize (Bt/non Bt/other GM maize). Section
3 referred to the specific measures to follow when cultivating Bt maize
(training, label recommendations on seed bags, use of refuge). In section 4
farmers were asked to submit their observations on fertility, use of fertilizers,
pests and diseases, weed and volunteer, use of herbicides, crop rotation and
crop history, use of crop protection products and proximate field
surroundings. Comparisons were always made to conventional maize.
Furthermore, space for the recording of additional observations was provided.
Section 5 related to post-harvest observations (level of control against target
pests, anything unusual, use of MON 810 for livestock). Other remarks, if
any, related to the cultivation of MON 810 could be added.
This questionnaire has been used by Monsanto and its licensee, Pioneer HiBred International, Inc.,which are the two main providers of MON 810 seeds
for cultivation in the EU.
The analysis of the 132 questionnaires collected in 2005 on the cultivation of
MON 810 maize did not indicate any adverse effect. The findings have been
provided to the European Commission and national competent authorities.
This first set of data is entered in a database which will be complemented year
after year with new entries constituted by responses to questionnaires.
The farm questionnaires are being distributed, completed and collated
annually. Reports will be prepared also on an annual basis, and in case of
adverse findings that require immediate risk mitigation, the results would be
reported to regulatory authorities immediately.
Learnings from 2005 experiences allowed Monsanto to develop a new and
improved version of the questionnaire that was used in Germany and elsewhere
in the 2006 season. (see Appendix 4). In particular, there has been a general
change from binary (Yes/No) to somantic answers with three levels (e.g. Less/
As usual/ More) to enable the farmers to specify the observed effects in two
different ways and to strengthen the analyses by bringing a higher statistical
power.
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MON 810 maize
May 2007
2) Peer reviewed publications on the safety of MON 810 and/or Cry1Ab
published in 2005 and 2006
An important source of information on MON 810 safety is the extensive
independent research that is performed by scientists with a wide range of
expertise, such as insect and microbial ecology, animal toxicology, molecular
biology or chemistry. More than 30 publications related to MON 810/Cry1Ab were
published in peer reviewed journals in 2005 and 2006. Those references related to
MON 810 or pure Cry1Ab were obtained by running a search using the search
engine ISI Web of KnowledgeTM (search terms: MON 810 or MON810;
Transgenic maize or corn; Bt maize or corn; Genetically modified maize or corn;
Cry1Ab and other).
These publications and other public research efforts reinforce our knowledge of
MON 810, its safety and commercial performance.
The data available, overall, indicate no detrimental effects of MON 810 and/or
Cry1Ab on human health or the environment.
The General Surveillance performed in 2006 will be submitted to the
Commission and national competent authorities in the coming months.
The GS plan for MON 810 cultivation is in line with both the EFSA guidance
document on monitoring (EFSA, 2006b) and the recommendations from the
Green Biotechnology Industry described by (Tinland et al., 2006).
APPLICABILITY OF THE MON 810 MONITORING PLAN TO THE
MONITORING REQUIREMENTS IN THE BVL ORDER
1. The environmental risk assessment
The Order issued by the BVL requires that the MON 810 monitoring plan takes
into account nine specific parameters (see italic text, below – translation by
Monsanto). We consider that the current monitoring plan for MON 810
cultivation in Germany, established on the basis of the conclusions of the
environmental risk assessment (e.r.a.) conducted according to the requirements
of Directive 2001/18/EC, are in accordance with the BVL Order for the following
reasons:
a) Release of maize grain that is able to germinate in the environment
(losses during harvesting, transportation and processing)
This hazard has been addressed in the environmental risk assessment in
Section 9.1 (Appendix 1). It is concluded that, as for conventional maize,
the likelihood of MON 810 adversely impacting the environment is
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MON 810 maize
May 2007
negligible, as it has shown no ability to be persistent or invasive and
these parameters are unaltered in MON 810 when compared to
conventional maize. In the unlikely event of the establishment of
MON 810 plants in the environment, the introduced trait would confer
only a limited selective advantage (protection from lepidopteran pests) of
short duration, narrow spatial context and with negligible consequences
for the environment. Hence the risk to the environment from MON 810
through increased persistence and invasiveness of this maize is
negligible.
b) Release of the Bt-toxin into the environment (for example via pollen,
silage, plant residues in the soil),
The extent of release of the Bt protein into the environment is one of
the components of the risk assessment (exposure) that was taken into
consideration while assessing the risk for each of the nine points of the
risk assessment under point “c) likelihood of the potential adverse
effect”. While the Bt protein will be present in the environment, its
selective insecticidal property and its low concentration are such that
it is unlikely to cause any adverse effect to the environment (e.g.: see
Appendix 1: 9.5.c.).
The safety of the Bt protein for human health and the environment has
been addressed in the risk assessment in each of the nine sections of
Appendix 1.
More specifically, the Cry1Ab protein expressed in MON 810 is similar
to the one present in microbial formulations that have been used safely
for nearly 45 years in commercial Bt-based insecticide sprays. There is
extensive information on the absence of non-target effects from the
Cry1Ab protein (Mendelsohn et al., 2003; Romeis et al., 2006).
To confirm and expand on the results produced for the microbial
products which contain the same Cry1Ab protein as produced in
MON 810, the potential impact of the Cry1Ab protein on non-target
organisms was assessed on several representative organisms. These
studies have previously been reported for MON 810 (Monsanto
Company, 1995) and are summarized in Table 1 (Appendix1). Studies
were conducted with the trypsin-resistant core of the Cry1Ab protein
because this is the insecticidally active portion of the Cry1Ab protein.
Non-target species that were tested include a) larval and adult
honeybees (Apis mellifera L.), which is a beneficial insect pollinator, b)
green lacewing larvae (Chrysopa carnea), a beneficial predatory insect;
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MON 810 maize
May 2007
c) Hymenoptera (Brachymeria intermedia), a beneficial parasite of the
housefly; d) the ladybird beetle (Hippodamia convergens), a beneficial
predaceous insect, and e) earthworms (Eisenia fetida), a representative
detritivorous species in the soil. In addition, leaf material of MON 810
plants was used in a non-target soil organism study using Collembola
(Folsomia candida). Due to the potential exposure of aquatic
invertebrates to maize pollen containing the Cry1Ab protein, a toxicity
test was also performed on daphnids (Daphnia magna). The U.S. EPA
has since determined that aquatic invertebrate tests are categorized as
supplemental studies since the likelihood of exposure of aquatic
animals including invertebrates to maize pollen is low (US EPA, 2002;
US EPA, 2005). The results of these non-target organism studies
showed that the mortality of non-lepidopteran insect species and three
other representative organisms exposed to the Cry1Ab protein at levels
in excess of potential environmental exposures did not significantly
differ from control mortality.
For the Bt proteins tested in laboratory assays to date, including
Cry1Ab, potentially significant adverse effects have been observed for
only a very few non-target species that are closely related to the target
species (Mendelsohn et al., 2003; Romeis et al., 2006). However, field
studies conducted over the past decade by industry and the academic
community, and reported in the peer-reviewed literature on registered
insect-protected crops that produce a variety of Cry1A proteins,
including Cry1Ab, have demonstrated that these crops have no adverse
effects on biodiversity, tested populations of natural enemies, and
other ecologically important non-target arthropods (U.S. and other
world areas: Daly and Buntin, 2005; Dively, 2005; Dively and Rose,
2003; Head et al., 2001; Head et al., 2005; Lozzia et al., 1998; Naranjo
et al., 2005; Naranjo, 2005a; Naranjo, 2005b; Orr and Landis, 1997;
Pilcher et al., 1997; Pilcher et al., 2005; Torres and Ruberson, 2005;
Whitehouse et al., 2005) (E.U.: (Arpas et al., 2005; Babendreier et al.,
2004; Bakonyi et al., 2006; Bourguet et al., 2002; Eckert et al., 2006;
Freier et al., 2004; Heckmann et al., 2006; Lang et al., 2004; Ludy and
Lang, 2006a; Ludy and Lang, 2006b; Meissle et al., 2005; Romeis et al.,
2004; Romeis et al., 2006; Toth et al., 2004; Vercesi et al., 2006; Vojtech
et al., 2005; Volkmar and Freier, 2003; Wandeler et al., 2002).
Importantly, even sensitive non-target lepidopteran species have been
shown to be exposed to lower levels of Cry1A proteins, when expressed
in Bt crops, compared to herbivores. These reduced levels of exposure
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MON 810 maize
May 2007
are too low to pose a significant risk to populations of the sensitive
non-target species (Hellmich et al., 2001; Pleasants et al., 2001). For
example, the impact of exposure to pollen containing Cry1A proteins
on lepidopteran species has been evaluated in a number of empirical
studies and several risk assessments (Mendelsohn et al., 2003), and
the risk has been shown to be negligible.
In conclusion, based on the well-characterised mode of action of the
Cry proteins, the selectivity of the Cry1Ab protein for certain
lepidopteran pests and the confirmation through studies showing no
adverse effects in diets, it is concluded that the potential for MON 810
to be hazardous to non-target organisms is negligible.
c)
Bt-toxin remaining in the soil of the cultivation areas; impacts on soil
organisms and soil functions,
been addressed in the risk assessment in Sections 9.1-9.9 and the effects
on biogeochemical processes has been specifically addressed in Section
9.8 of the e.r.a. (Appendix 1).
The Cry1Ab protein was shown to degrade rapidly in soil, which
confirms the absence of adverse effects on soil microorganisms. The
degradation rate of the Cry1Ab protein was assessed by measuring the
decrease in insecticidal activity of MON 810 tissue incubated in soil.
The Cry1Ab protein, as a component of the maize tissue, had an
estimated DT50 (time to 50% reduction of bioactivity) and DT90 (time to
90% reduction of bioactivity) of 1.6 and 15 days, respectively (Sims and
Holden, 1996). This measured rate of degradation in soil is comparable
to that reported for the Btk protein in genetically modified cotton
(Palm et al., 1994) and to the degradation rate reported for microbial
Bt products (Pruett et al., 1980; West, 1984; West et al., 1984). This
rapid degradation strongly supports the lack of exposure of Cry1Ab on
non-target organisms involved in the decomposition function, and on
soil-dwelling organisms in general. More recently, Dubelman et al.,
2005 showed that Cry1Ab protein does not accumulate or persist in the
environment after 3 years of continuous use.
Throughout its lifecycle in the field, MON 810 interacts with a
spectrum of non-target organisms that are involved in the
biogeochemical processes of decomposition and nutrient recycling in
the soil. As biogeochemical processes are exceedingly complex, such
processes are best understood at a macroscopic, system level than at
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specific, organism level. The main functional groups of non-target
organisms that are relevant to the assessment of potential adverse
effects on biogeochemical processes include decomposers of plant
material and organic substances, and primary consumers feeding on
organic debris (detritivores).
Primary consumers typically are macro-organisms that feed on the
detritus, i.e. the organic debris resulting from decomposition of plant
material. They further reduce the size of the detritus particles through
partial digestion, and, after defecation of the particles, thereby
enhance further decomposition. Detritivores also provide aeration to
the soil or litter material, thereby improving the oxygen content in the
soil and increasing the respiration of decomposers. Important
examples of detritivores are springtails (Collembola), millipedes and
annelid worms. Populations of soil-borne consumers are affected by
plant genotype, by tillage practices, environmental conditions, previous
history of crops grown and the application of pesticides and fertilisers.
The Cry1Ab protein expressed in MON 810 was demonstrated to have
negligible risk for the environment through direct or indirect
interactions with non-target organisms, including representative
detritivorous organisms that are involved in the decomposition
function in the soil.
Decomposers include bacteria and fungi (saprophytes) that break down
dead and decaying material, such as residual crop plant material, fresh
litter remaining after harvest of the crop, smaller detritus from more
advanced decomposition, and humus. Dead and decaying plant
material contains important nutrients, e.g. carbon which is released as
carbon dioxide as a result of microbial respiration, and nitrogen which
is recycled by a range of soil bacteria. Bacterial and fungal populations
are critical to maintaining soil health and quality. Soil microbial
communities that mediate biogeochemical processes are highly
complex and are often characterized by high microbial diversity (Tiedje
et al., 1999). However, the diversity and abundance of these organisms
and hence their microbial processes are significantly affected by biotic
factors (community characteristics and dynamics), abiotic factors (soil
structure, clay type, moisture capacity, environmental conditions, pH)
and soil use (crop, tillage practices, history of previously grown crops).
Agricultural practices such as fertilization and cultivation techniques
may also have profound effects on soil microbial populations, species
composition, colonization, and associated biochemical processes
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(Alexander, 1961). Consequently, significant variation in microbial
populations is expected in the agricultural environment.
Although the Cry1Ab protein present in decaying MON 810 material is
considered to be a newly expressed protein in maize, it is not a novel
protein in the soil. The cry1Ab gene which was used in this genetically
modified maize was derived from the genome of a common soil
bacterium Bacillus thuringiensis subsp. kurstaki.
The toxic mechanism of Cry1Ab protein has been thoroughly
characterised (see Monsanto Company, 1995) and was found to be
extremely specific to larvae of certain lepidopteran insect pests.
Consequently, the potential for activity of this protein towards
microorganisms is negligible. The rapid degradation of Cry1Ab in soil
strongly supports the lack of exposure of Cry1Ab on non-target
organisms involved in the decomposition function and on soil-dwelling
organisms in general.
Finally, extensive commercial experience with the commercialisation of
various Cry1Ab-expressing insect-protected crops has not revealed any
adverse or undesirable effects on biogeochemical processes or soil
fertility.
It is concluded in the risk assessment that the probability of adverse
effects on biogeochemical processes, caused by the interaction of
MON 810 with target and non-target organisms in the soil, is
negligible, since it is highly unlikely that there is any difference
between MON 810 and conventional maize with respect to its direct
influence on soil nutrient levels and key processes. Furthermore, it is
also highly unlikely that the direct or indirect interaction between this
maize and decomposers or detritivores in the receiving environment
would cause any immediate or delayed, or direct or indirect, adverse
effects on the decomposition and nutrient recycling functions in the
soil.
d) Effects on non-target organisms on the cultivation areas and in affected
eco-systems near the cultivation areas
been addressed in the risk assessment in Sections 9.1-9.9 (Appendix 1)
and the effects on non target organisms has been specifically addressed
in Section 9.5. of the e.r.a. (Appendix 1) and are summarized above
under point b).
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It is concluded in the risk assessment that there is negligible risk of
harmful effects of MON 810 on non-target organisms (vertebrates and
invertebrates), either through direct or indirect interactions with this
maize or through contact with the Cry1Ab protein. Higher trophic
interactions between non-target organisms would also not be
negatively affected. Therefore, any risks of significant indirect effects
on the population levels of non-target organisms in the receiving
environment or their functioning in below- and above-ground
ecosystems in the vicinity of the crop are equally negligible.
e)
Long-term and large-scale effects on bio-diversity
and the effects on non target organisms processes has been specifically
addressed in Section 9.5. of the e.r.a. (Appendix 1) and are summarized
above under point b).
f)
Issue of transgenic organisms remaining in organisms
environmental media (by persistence and accumulation)
and
The potential persistence and accumulation of the Cry1Ab protein in
organisms and environmental media is one of the component of the
risk assessment (exposure) that was taken into consideration under “c)
likelihood of the potential adverse effect” while assessing the risk for
each of the nine points of the environmental risk assessment
(Appendix 1).
While the Bt protein will be present in the environment, its selective
insecticidal property and its low concentration are such that it is
unlikely to cause any adverse effects on the environment as explained
in Section 9.5.c. (Appendix 1) and above summarized under point b).
The Cry1A proteins bind specifically to receptors on the midgut of
lepidopteran insects (Hofmann et al., 1988a; Hofmann et al., 1988b;
Van Rie et al., 1989; Van Rie et al., 1990; Wolfersberger et al., 1986)
and have no deleterious effect on beneficial or other non-target insects,
including predators and parasitoids of lepidopteran insect pests or
honeybees (Apis mellifera) (Cantwell et al., 1972; Flexner et al., 1986;
Krieg and Langenbruch, 1981; Melin and Cozzi, 1990; US EPA, 2000;
Vinson, 1989). Selectivity based on the mode of action is a key factor in
the safety of Cry proteins for non-target organisms such as fish, birds,
mammals and non-target invertebrates.
Page 14 of 33
MON 810 maize
May 2007
g) Development of secondary pests
and the impact of the specific cultivation, management and harvesting
techniques has been specifically addressed in Section 9.9. of the risk
assessment. (Appendix 1). Specifically, based on all the evidence, it is
concluded that “the environmental impact of farming practices to
grow MON 810 in the E.U. is considered no different from any other
maize.”
The potential of an insect pest control technology to induce the
development of secondary pests in a crop ecosystem depends largely on
the potential for interruption of the biological communities
(particularly natural enemy complexes) by the control method. In
general, broader target spectrum of the control measures result in
more severe interruption of biological communities in the crop
ecosystem (Kogan 1998). Development (or induction) of secondary
pests by consistent use of broad-spectrum insecticides has long been
documented in many crop ecosystems such as potato ( e.g., Reed et al.
2001), cotton, and apple orchards (see reviews in Kogan 1998).
However, we do not expect this to be the case with MON 810 maize in
the E.U. or elsewhere, because of its narrow-spectrum insecticidal
properties.
In contrast to broad-spectrum insect pest control technologies such as
chemical insecticides, MON 810 maize expresses the Cry 1Ab protein
that is only active against the selected lepidopteran pests (primarily
European corn borers), and has much less potential to interrupt
biological communities of the maize ecosystem. This conclusion is
supported by results of numerous non-target organism risk assessment
studies (Section 9.5 in Appendix 1), as well as studies on fauna (e.g.,
Candolfi et al. 2004). The potential for induction or development of
secondary pests, if any, would therefore not be an intrinsic
consequence of the direct use of MON 810. Rather, shifts in pest
spectrum if they were to occur would be a consequence of replacing the
other control measures, such as those insecticide sprays against
lepidopteran maize pests that are currently approved for use in maize
production and which may still be used with MON 810 maize.
Importantly for the risk assessment, MON 810 is not different from
conventional maize, except for the introduced lepidopteran-protection
Page 15 of 33
MON 810 maize
May 2007
trait. All agronomic practices currently used to grow maize in the E.U.
remain applicable for growing MON 810 and no new or specific
techniques for the cultivation, management and harvesting of MON
810 are necessary. In other words, traditional crop rotational practices,
planting regimes for maize, techniques for soil preparation (tillage)
and all technical equipment remain applicable. Similarly, no new or
specific crop management techniques are required for MON 810. All
the conventional best management techniques to cultivate maize
remain at the farmer’s discretion. Finally, no changes in harvesting
techniques are required. Traditional harvesting equipment as well as
post-harvest storage techniques and conditions remain applicable.
In summary, based on the characteristics of MON 810, the risk
assessment information submitted (Appendix 1, Sections 9.1-9.9) and
its proposed use in maize production, there is no reason to believe that
secondary pests are likely to develop that will pose new or unique risks
to the environment in the E.U. or elsewhere. This is because the
development of secondary pests is associated with more profound
changes in insect pest management practices than will occur with
MON 810 maize. Growers will have available and may choose to use
other approved insect pest management practices with MON 810 as
part of their maize production system. These approved practices may
affect the nature and spectrum of pests that they will need to manage.
Importantly, the principle goal of
management technique will
continue to be of the removal of harmful insect pests from the field in
order to achieve optimal yield of the crop, and these are not different
between MON 810 and any other maize. Therefore, cultivation of
MON 810 instead of conventional maize does not change any basic
management technique in maize as such, but gives growers more
flexibility to apply the existing tools for management, while creating at
the same time new opportunities to grow maize in a more sustainable
way (e.g. reduced tillage or integrated pest management).
h) Modifications of pesticide applications (type of pesticide, volume,
frequency and point in time)
been addressed in the risk assessment in Sections 9.1-9.9 (Appendix 1).
The impact on agricultural practices is detailed in Section 9.9. of the
e.r.a. (Appendix 1).
Page 16 of 33
MON 810 maize
May 2007
In comparison to conventional maize, the altered management
practices employed for the production of MON 810 maize do not have
adverse effects on the environment. This has been established from the
risk assessment and from practical experiences with the cultivation of
MON 810 in the EU and elsewhere.
It has been demonstrated that the production of MON 810 positively
impacts current agronomic practices in maize and provide benefits to
farmers and the environment in the EU. The benefits of planting
insect-protected maize include: 1) a reliable means to control the target
lepidopteran maize pests; 2) control of target insects while maintaining
beneficial species; 3) reduced use of chemical insecticides (Rice and
Pilcher, 1999); 4) reduced operator exposure to chemical pesticides; 5)
good fit with integrated pest management (IPM) and sustainable
agricultural systems; 6) reduced fumonisin mycotoxin levels in maize
kernels (Masoero et al., 1999; Munkvold et al., 1999); and 7) no
additional labour or machinery requirements, allowing both large and
small growers to maximize hybrid yields. MON 810 can offer the
above-mentioned agronomic and environmental benefits, and therefore
also societal benefits.
i)
Impacts on food chains and webs.
been addressed in the risk assessment in Sections 9.1-9.9 (Appendix 1).
The impact on food chains and webs are more specifically addressed in
Sections 9.5. and 9.8. of the e.r.a. (Appendix 1) and summarized above
under point b) and c), respectively.
It was concluded in the risk assessment that there is negligible risk of
harmful effects of MON 810 on non-target organisms (vertebrates and
invertebrates), either through direct or indirect interactions with this
maize or through contact with the Cry1Ab protein. Higher trophic
interactions between non-target organisms would also not be
negatively affected. Therefore, any risks for significant indirect effects
on the population levels of non-target organisms in the receiving
environment or their functioning in below- and above-ground
ecosystems in the vicinity of the crop are equally negligible.
In conclusion, the environmental risk assessment submitted in the
application for the renewal of the MON 810 cultivation approval in the
EU confirms the conclusions of the previous risk assessment conducted
according to Directive 90/220/EEC. Furthermore, it addresses the nine
Page 17 of 33
MON 810 maize
May 2007
specific parameters given in the BVL Order and establishes that casespecific monitoring of these parameters is not warranted in the case MON
810 cultivation in Germany. This is in line with the overall conclusion of
the e.r.a that concludes that the deliberate release of MON 810 into the
environment consequence is unlikely to cause any risk to the
environment, and therefore that no case specific monitoring should be
implemented.
However, as discussed and detailed in the section “State-of-the-art
monitoring plan for MON 810”, a general surveillance plan was developed
to identify the occurrence of adverse effects of MON 810 which were not
identified or anticipated in the e.r.a. The following section will detail how
the parameters a) to i), cited in the BVL Order, are covered from a
perspective of general surveillance.
2. General Surveillance for MON 810
The objective of Directive 2001/18/EC is to protect “human health and the
environment”. Although it is difficult to define general monitoring parameters,
concrete and informative monitoring characters for general surveillance can be
derived from more specific protection goals and their areas of potential impact
covering the term “human health and the environment” [Wilhelm, 2003 #3965].
Protection goals are:
• Ecological systems and biodiversity
• Soil function
• Sustainable agriculture
• Plant health
• Human and animal health
The monitoring parameters a) to i) cited in the BVL Order are covered by those
protection goals that are the basis of the General Surveillance for MON 810
cultivation (Appendix 3), and more specifically in the “Questionnaire to Farmer”
(Appendix 4). This is one of the three pillars of MON 810 general surveillance
(GS), in addition to Monsanto stewardship activities and analysis of the relevant
literature.
More specifically, the following section describes how the farmer questionnaire
component of General Surveillance addresses points a) to i) of the BVL Order,
but it is important also to note that stewardship and literature analysis also
contribute to this analysis and that all the tools together have so far confirmed
the outcome of the risk assessment.
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MON 810 maize
May 2007
The GS program that will be implemented during the 2007 contains a
questionnaire to farmers that allows to assess points a) - i) of the BVL
Order:
a) Release of grain maize that is able to germinate into the environment
(losses during harvesting, transportation and processing)
The potential presence of volunteers is addressed specifically by a
question under Section 3.2. of the questionnaire (Appendix 4). Other
questions in this Section 3.2. address the general characteristics of
MON 810 with comparison to conventional maize. This section
provides the means to detect any unanticipated “risk of persistence or
invasiveness”, considered negligible following the e.r.a. (Section 9.1,
Appendix 1).
b) Release of the Bt-toxin into the environment (for example via pollen,
silage, plant residues in the soil)
Existing General Surveillance monitoring will assess potential adverse
effect on human health and the environment. Potential adverse effects
associated to the release of the Bt-toxin in the environment are
addressed by the farmer questionnaire, as described in the following
sections.
c) Bt-toxin remaining in the soil of the cultivation areas; impacts on soil
organisms and soil functions
From a general surveillance standpoint, the analyses the crop
performance can be considered as a good indicator of the soil quality
and functionalities. The questions that are addressed in Section 3 of
the questionnaire compare MON 810 growth characteristics with those
of conventional maize. In addition, the farmer is asked, under Section
1.3, to assess the quality of the soil in his fields with a high level of
detail. Any deterioration in soil quality over the years and across
locations can be monitored thanks to the accumulation of spatial and
temporal data. If such deterioration were to occur, an in depth
investigation will be carried on to understand the cause.
d) Effects on non-target organisms on the cultivation areas and in affected
eco-systems near the cultivation areas
Detection of critical changes in non-target organisms in a maize field is
addressed under Question 3.7, which assesses the occurrence of
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MON 810 maize
May 2007
wildlife in fields where MON 810 are grown (mammals, birds, insects)
as compared to conventional maize. Farmers are asked to describe and
specify unusual observations.
e) Long-term and large-scale effects on bio-diversity
A GS database is developed to manage the data from farm
questionnaires. It will also be connectable to data from other sources.
If a potential adverse effect is identified, the party placing the GM
plant on the market can gather additional data to understand whether
this adverse effect is associated with the GM plant. The key for data
pooling from different sources should be their temporal and spatial
coordinates. Therefore, all data sets are identified by their origin – the
date and location of survey. A matching of spatiotemporal coordinates
of datasets coming from different sources allows, for example,
assessing whether an observed negative effect on plant development
reported in a subset of questionnaires for a certain region, can be
related to a higher occurrence of plant diseases in that region observed
by a public plant protection network.
f) Issue of transgenic organisms remaining in organisms
environmental media (by persistence and accumulation)
and
Potential adverse effects associated with the presence of material of
GM origin in organisms or in the environmental media are addressed
by the questionnaire to farmers. If the persistence of MON 810 would
be translated into an adverse effect, it would likely to be detected by
the GS (see other sections in this chapter).
g) Development of secondary pests
Surveying pest development in its field is critical to any farmer since it
as the potential to impact directly upon their revenue. The
questionnaire to farmers requires extensive characterization of
diseases, pest, and weed pressure in MON 810 fields as compared to
conventional maize (Questions in Sections 3.3.-3.6). Should it occur,
the GS will therefore allow to detect the development of secondary
pests. The information that is generated across areas and years will be
stored in a database according to temporal and spatial coordinates.
This will reveal any trends of pest development, and will allow
comparison with data generated by existing networks such as Plant
Protection or meteorology services. The processed information will
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MON 810 maize
May 2007
provide a very powerful tool that will help in optimizing integrated
pest management strategy.
h) Modifications of pesticide applications (type of pesticide, volume,
frequency and point in time)
Experience has demonstrated that the use of Bt crops is accompanied
by a reduction of pesticide applications. The Questions under Section
3.1 of the questionnaire to farmers address changes in agricultural
practices in MON 810 fields compared with conventional maize. The
modification of pesticide regimes (insecticides, pesticides, fungicides) is
specifically addressed. Open questions allow the farmers to explain
any changes in pesticide use. In common with other farmer responses,
accumulation of data identified with their temporal and spatial
coordinates will allow detection of trends and correlation with
particular conditions. Note also that cross checking this information
with the one detailed in the previous point on secondary pest
development (if any) could give insight into the origin of such
development.
i) Impacts on food chains and webs
Several sections of the farmer questionnaire would reveal adverse
effects of MON 810 on food chains and webs as compared to
conventional maize. The most relevant are questions in Section 3.7,
that assesses the occurrence of wildlife in environments where
MON 810 is grown, and questions in Sections 3.3.-3.6. that address the
development of secondary pests (which could be associated to trophic
levels of predation). In addition, adverse effects on livestock consuming
MON 810 maize is assessed by questions in Section 3.8. Answers to the
other questions of the questionnaire, such as those which assess the
agronomic characteristics of MON 810 (questions under Section 3) are
also relevant, since the equivalence of MON 810 to conventional maize
is a determinant in the presence of all organisms that are associated
either directly or indirectly to maize.
Questionnaires to farmers have been established as an essential tool for the
general surveillance of MON 810. It permits the comparison of information
within the same questionnaire, and across regions and years, and therefore
constitutes a powerful tool to detect adverse effects as well as general trends at
early stages. The data are stored in the way that they could be compared to
other network in place.
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MON 810 maize
May 2007
Any recorded observations of adverse findings that are linked to the cultivation
and/or use of MON 810, which come to the attention of the party placing the GM
plant on the market will be reported immediately. It will receive careful analysis
in real time, and, if necessary, remediation action. Annual general surveillance
reports will be sent to the European Commission, which will distribute to all
Competent Authorities in the E.U. General Surveillance reports will be prepared
on an annual basis, except in case of adverse findings that need immediate risk
mitigation, which will be reported immediately.
Since monitoring of GM plants is a new topic and a creative process, the
monitoring plan and especially the questionnaires can be improved based on
experience from year to year. In this respect, the questionnaire that has been
submitted as part of the renewal application for the renewal of MON 810
cultivation authorization in Europe was considerably improved with respect to
its initial format used during the 2005 season. Those improvements have been
made, firstly on the quality of the questions based on the feed back gathered
from the interviewers, the farmers and the analyst, secondly on the design of the
data collection in order to facilitate the data storage, interpretation and
statistical analysis.
3. Comments to the publications submitted as reasoning for the
proposed safeguard measure on MON 810 maize
In Part II (“Begründung”), the BVL Order cites a number of publications as new
evidence pointing to potential effects of MON 810 on a) non-target organisms
and b) the soil environment. These points have been addressed above in the
environmental risk assessment but are re-discussed in the following section in
light of the specific papers brought up in the Order.
a)
Risk for non-target organisms
In this section, the justification discusses adverse effects of MON 810 on
NTOs due to the potential for Cry1Ab to pass up the food chain via the
plant, cites the review by (Lovei and Arpaia, 2005) that assesses the impact
of transgenic plants on natural enemies and asserts potential risk to nontarget butterflies exposed to the Cry1Ab protein in pollen from MON 810.
•
Comments on potential for Cry1Ab to pass up the food chain:
References cited in the justification for the BVL Order indicate that
higher trophic levels (e.g., predators and parasitoids) may be
exposed to Bt Cry proteins produced in transgenic plants via trophic
interactions (Harwood et al., 2005; Obrist et al., 2006; Zwahlen and
Andow, 2005). This type of indirect exposure to a toxin or
Page 22 of 33
MON 810 maize
May 2007
insecticide (i.e., via tri-trophic interactions) is often referred to as
secondary exposure, the degree of which is subject to the influences
of various ecological and behavioral factors such as the fate and/or
form of the toxin in the prey’s digestive system and/or tissues,
feeding behavior of the prey, as well as predators and parasitoids.
While there is the potential for predators and parasitoids to be
exposed to Bt Cry proteins produced in Bt crops (see Appendix 1),
several studies have shown that the magnitude of secondary
exposure is generally much less than that of primary exposure via
direct feeding on Bt protein-producing plants (Dutton et al., 2002;
Head et al., 2001; Raps et al., 2001). This is largely because the
maximum amount of Bt protein that an organism (such as an insect
herbivore) may carry is limited by the total volume of its alimentary
canal, the rate of intake of Bt protein-producing plant tissue, as
well as its susceptibility to Bt proteins (Harwood et al., 2006). Thus,
from the perspective of risk assessment, the secondary exposure
risk of Bt crops to the third or higher trophic levels (predators and
parasitoids), if any, would be much less than to those trophic levels
that directly feed on Bt plants (i.e., herbivores). More importantly,
many laboratory and field studies have clearly demonstrated that
the Cry1Ab protein expressed in MON 810 has little toxicity (or
hazard) towards non-lepidopteran (non-target) organisms including
diverse groups of arthropod predators and parasitoids (see
Appendix 1).
•
Comments on the review by Lovei and Arpaia (2005): Lovei and
Arpaia (2005) evaluated 26 laboratory studies, which tested pure
protein, spiked prey or plant material associated with or derived
from GM insecticidal plants for effects on predatory NTOs. Of the
26 studies reviewed, 9 were based on GM plants or proteins, such
as protease inhibitors and lectins, which were never
commercialized, have no relationship to Cry1Ab in terms of their
mode of action, and are generally known in the scientific literature
to have broad toxic effects against a number of different animal
taxa. Another 7 studies were performed on either Cry1Ac or Cry3A
proteins or plants which contained these proteins. Only 10 of the 26
studies were conducted on some form of Cry1Ab, e.g., derived from
events Bt11, 176 or MON 810 or in one case uncharacterized test
material used in experimental rice. Of these 10 studies, 5 showed
no effects on NTOs and five produced effects. In all cases where an
Page 23 of 33
MON 810 maize
May 2007
effect was observed it was either indirect, i.e., an effect due to
predators feeding on poisoned or weakened target pests, or to
problems associated with the test system, e.g., the lacewing test
system (Romeis et al., 2004).
Lovei and Arpaia (2005) also evaluated 18 laboratory studies, which
tested pure protein, spiked prey or plant material associated with
or derived from GM insecticidal plants for effects on parasitoid
NTOs. Of the 18 studies reviewed, 13 were based on GM plants or
proteins, such as protease inhibitors and lectins, which as stated
above were never commercialized, have no relationship to Cry1Ab
in terms of their mode of action, and are generally known in the
scientific literature to have broad toxic effects against a number of
different animal taxa. Another 3 studies were performed on either
Cry1Ac or Cry3A proteins or plants which contained these proteins.
Of the 2 studies that used Cry1Ab, one was based on maize event
CG176 and the other uncharacterized test material used in
experimental rice. The experiments using the event CG176
material showed a tritrophic effect which was likely due to
parasitoid interactions with poisoned or weakened target pests. No
effects were observed with the rice material. Importantly, these
laboratory test systems have been questioned by experts in this
field (Romeis et al., 2006b).
•
Comments on potential risk to non-target butterflies: The Cry1A
family of toxins are active against lepidopteran insects. MON 810
maize was developed to specifically target insect pests in the order
Lepidoptera. The toxicity of the Cry1Ab protein found in MON 810
has been assessed against at least 10 lepidopteran families
comprised of at least 20 lepidopteran species. Toxicity values for the
Cry1Ab protein against lepidopteran insects range from 0.0033
µg/ml for Monarch butterfly (Danaus plexippus; Danaidae) to 3.6
µg/ml for European corn borer (Hellmich et al., 2001; MacIntosh et
al., 1990). Based on protein toxicity assessments, the Monarch
butterfly is considered to be one of the most sensitive lepidopteran
species to the Cry1Ab protein. Because it is not possible to test the
Cry1Ab protein against all lepidopteran insects, especially those
that are endangered or threatened or otherwise protected, the use
of a sensitive species such as Monarch butterfly provides a suitable
surrogate species for risk assessment of potential effects on other
Page 24 of 33
MON 810 maize
May 2007
lepidopterans (Mendelsohn et al., 2003; Romeis et al., 2006a; Wolt
et al., 2005).
A conservative estimate of the concentration of Cry1Ab in pollen
from MON 810 is <0.09 ug/g (Hellmich et al., 2001). This estimate
would translate to an activity against Monarch butterfly of around
>366 pollen grains/cm. Experimental results show that the actual
dose of pollen from MON 810 required for an effect is in excess of
1,000 pollen grains/cm.
Sears et al. (2001) developed a risk assessment model for Monarch
butterfly that used these toxicity data and incorporated estimates
of exposure based on the temporal and spatial distribution of maize
pollen, the Monarch’s host plant, as well as the presence and
susceptibility of Monarch butterfly larvae. Since the Monarch’s host
plant (Asclepias syriaca) occurs in maize fields at the time of maize
pollen shed, this assessment represents a worst case scenario for
susceptible butterflies. Their assessment concluded that there was
minimal risk to Monarch butterflies due to MON 810 and that the
risk was determined predominantly by exposure. Thus, an
assessment of the potential risk of MON 810 to other lepidopteran
insects, including threatened or endangered species can be provided
based on their potential exposure to pollen from MON 810 even
assuming a worst case toxicity scenario.
b)
Effects on soil organisms
References used in the assessment presented in the BVL Order to justify
the persistence of Cry1Ab from MON 810 are primarily based on model
laboratory studies (Creccio and Stotzky, 2001) or studies where the
concentration of the protein is measured in bagged corn tissues that are
buried in the soil (Zwahlen et al., 2003), rather than by measuring the
concentration of the protein in the soil itself. However, a large scale multiyear study that was designed to monitor the persistence and accumulation
of the Cry1Ab protein in five corn-growing areas of the United States
(Illinois, Iowa, Maryland, Wisconsin and South Dakota) was apparently not
considered in this assessment.
This field study, which has a higher environmental relevance than those
cited in the Order, showed that there was no persistence or accumulation of
the Cry1Ab protein in any of the sites where MON 810 was grown
consecutively for three or more years (Dubelman et al., 2005).
Page 25 of 33
MON 810 maize
May 2007
The MON 810 field study was performed in a wide geographic area and
under real agronomic and environmental conditions. The soil collection sites
were chosen to include many different soil characteristics, e.g., high clay (up
to 29%), pH variation (pH 4.7 to pH 7.8), organic matter content (1.6% to
5.1%) and cation-exchange capacity (8.6 to 31 meq/100g). Soil samples were
collected in plots where Bt-corn was grown for at least three consecutive
years, and control soils were also collected from nearby non-Bt crops. The
presence of the Cry1Ab protein in soil was assessed by a sensitive insect
bioassay (European corn borer) that statistically compared the insect
toxicity of soils collected from Bt and non-Bt fields.
The lack of persistence reported in the MON 810 field study (Dubelman et
al., 2005) demonstrates that laboratory studies (e.g., Stotzky) and surrogate
tissue studies (e.g., Zwahlen) are often not good indicators of environmental
persistence. The laboratory studies overestimate soil persistence because it
is not possible to properly model under laboratory conditions all the realworld environmental conditions that may contribute to dissipation
(sunlight, rain, microbial biomass and activity, temperature changes, etc.).
Further, several researchers have no observed no adverse effects of Bt
proteins on the soil microflora (Koskella and Stotzky, 2002; Saxena and
Stotzky, 2001).
In conclusion, a detailed review of the publications submitted in the BVL Order
as reasoning for a potential safeguard clause in no way alter the established
conclusions on the safety of MON 810 for human health and the environment, as
presented above in the environmental risk assessment. The weight of evidence
from laboratory and confirmatory field studies, as well as over 10 years of
commercial production in the U.S. and other countries, including the EU since
1998, support the conclusion that the cultivation of MON 810 poses negligible
risk to NTOs, and the environmental risk of MON 810 is considered to be
negligible compared to conventional maize.
Page 26 of 33
MON 810 maize
May 2007
REFERENCES
Alexander, M. (1961) Introduction to soil microbiology. John Wiley and Sons.
Arpas, K., Toth, F. and Kiss, J. (2005) Foliage-dwelling Arthropods in Bttransgenic and Isogenic Maize: A comparison through spider web
analysis. Acta Phytopathologica et Entmologica Hungarica, 40, 347-353.
Babendreier, D., Kalberer, N., Romeis, J., Fluri, P. and Bigler, F. (2004) Pollen
consumption in honey bee larvae: a step forward in the risk assessment
of transgenic plants. Apidologie, 35, 293-300.
Bakonyi, G., Szira, F., Kiss, I., Villanyi, I., Seres, A. and Szekacs, A. (2006)
Preference tests with collembolas on isogenic and Bt maize. Eur. J. Soil
Biol., 42, S132 - S135.
Bourguet, D., Chaufaux, J., Micoud, A., Delos, M., Naibo, B., Bombarde, F.,
Marque, G., Eychenne, N. and Pagliari, C. (2002) Ostrinia nubilalis
parasitism and the field abundance of non-target insects in transgenic
Bacillus thuringiensis corn (Zea mays). Environ. Biosafety Res., 1, 49-60.
Brookes, G. (2007) The benefits of adopting genetically modified, insect resistant
(Bt) maize in the European Union (EU): first results from 1998-2006
plantings. www.pgeconomics.co.uk, 1-39.
Candolfi, M.P., Brown, K., Grimm, C., Reber, R. and Schmidli, H. (2004) A
faunistic approach to assess potential side effects of genetically modified
Bt corn on non-target arthropods under field conditions. Biocontrol
Science and Technology, 14, 129-170.
Cantwell, G.E., Lehnert, T. and Fowler, J. (1972) Are biological insecticides
harmful to the honey bee? American bee journal, 294-296.
Creccio, C. and Stotzky, G. (2001) Biodegradation and insecticidal activity of the
toxin from Bacillus thuringiensis subsp.kurstaki bound on complexes of
montmorillonite-human acids-A1 hydroxypolymers. Soil Biology and
Biochemistry, 33, 573-581.
Daly, T. and Buntin, G.D. (2005) Effect of Bacillus thuringiensis transgenic corn
for Lepidopteran control on nontarget arthropods. Environ. Entomol., 34,
1292-1301.
Dively, G.P. (2005) Impact of transgenic VIP3A x Cry1Ab Lepidopteran-resistant
field corn on the nontarget arthropod community. Environ. Entomol., 34,
1267-1291.
Page 27 of 33
MON 810 maize
May 2007
Dively, G.P. and Rose, R. (2003) Effects of Bt transgenic and conventional
insecticide control on the non-target natural enemy community in sweet
corn. Proceedings of the 1st International Symposium on Biological
Control of Arthropods., 265-274.
Dubelman, S., Ayden, B.R., Bader, B.M., Brown, C.R., Jiang, C. and Vlachos, D.
(2005) Cry1Ab Protein does not persist in soil after 3 years of sustained
Bt Corn use. Environm. Entomol., 34, 915-921.
Dutton, A., Klein, H., Romeis, J. and Bigler, F. (2002) Uptake of Bt-toxin by
herbivores on transgenic maize and consequences for the predator
Chrysoperla carnea. Ecol. Entomol., 27, 441-447.
Eckert, J., Schuphan, I., Hothorn, L.A. and Gathmann, A. (2006) Arthropods on
maize ears for detecting impacts of Bt maize on non target organisms.
Environmental Entomology, 35, 554-560.
EFSA. (2006a) New chapter 11.4: general surveillance of unanticipated adverse
effects of the GM plant. In the EFSA Guidance document of the Scientific
Panel on genetically modified organisms for the risk assessment of
genetically modified plants and derived food and feed, 1-11.
EFSA. (2006b) Opinion of the Scientific Panel on genetically modified organisms
on the post-market environmental monitoring (PMEM) of genetically
modified plants. Question No EFSA-Q-2004-061, 319, 1-27.
Flexner, J.L., Lighthart, B. and Croft, B.A. (1986) The effects of microbial
pesticides on non-target, beneficial arthropods. Agriculture, ecosystems
and environment, 16, 203-254.
Freier, B., Schorling, M., Traugott, M., Juen, A. and Volkmar, C. (2004) Results
of a 4-year plant survey and pitfall trapping in Bt maize and
conventional maize fields regarding the occurrence of selected arthropod
taxa. IOBC/wprs Bulletin, 27.
Harwood, J.D., Samson, A. and Obrycki, J.J. (2006) No evidence for the uptake
of Cry1Ab Bt-endotoxins by the generalist predator Scarites
subterraneus (Coleoptera: Carabidae) in laboratory and field
experiments. Biocontrol Science and Technology, 16, 377-388.
Harwood, J.D., Wallin, W.G. and Obrycki, J.J. (2005) Uptake of Bt endotoxins by
nontarget herbivores and higher order arthropod predators: molecular
evidence from a transgenic corn agroecosystem. Molecular Ecology, 14,
2815-2823.
Page 28 of 33
MON 810 maize
May 2007
Head, G., Brown, C.R., Groth, M.E. and Duan, J.J. (2001) Cry1Ab protein levels
in phytophagous insects feeding on transgenic corn: implications for
secondary exposure risk assessment. Entomologia Experimentalis et
Applicata, 99, 37-45.
Head, G., Moar, W., Eubanks, M., Freeman, B., Ruberson, J., Hagerty, A. and
Turnipseed, S. (2005) A multiyear, large-scale comparison of arthropod
populations on commercially managed Bt and non-Bt cotton fields.
Environ. Entomol., 34, 1257-1266.
Heckmann, L.H., Griffiths, B., Caul, S., Thomson, J., Pusztai-Carey, M., Moar,
W.J., Andersen, M.N. and Krogh, P.H. (2006) Consequences for
Protaphorura armata (Collembola: Onychiuridae) following exposure to
genetically modified Bacillus thuringiensis (Bt) maize and non-Bt maize.
Environmental Pollution, 142, 212-216.
Hellmich, R.L., Siegfried, B.D., Sears, M.K., Stanley-Horn, D.E., Daniels, M.J.,
Mattila, H.R., Spencer, T., Bidne, K.G. and Lewis, L.C. (2001) Monarch
larvae sensitivity to Bacillus thuringiensis-purified proteins and pollen.
Proc. Natl. Acad. Sci., 98, 11925-11930.
Hofmann, C., Luthy, P., Hutter, R. and Pliska, V. (1988a) Binding of the delta
endotoxin from Bacillus thuringiensis to brush- border membrane
vesicles of the cabbage butterfly (Pieris brassicae). Eur J Biochem, 173,
85-91.
Hofmann, C., Vanderbruggen, H., Hoefte, H., Van Rie, J., Jansens, S. and Van
Mellaert, H. (1988b) Specificity of Bacillus thuringiensis delta-endotoxins
is correlated with the presence of high-affinity binding sites in the brush
border membrane of target insect midguts. Proc. Natl. Acad. Sci. USA,
85, 7844-7848.
Kogan, M. (1998) Integrated pest management: historical perspective and
contemporary development. Annual Review of Entomology, 43, 243-270.
Koskella, J. and Stotzky, G. (2002) Larvicidal toxins from Bacillus thuringiensis
subspp. kurstaki, morrisoni (strain tenebrionsis), and israelensis have no
microbiocidal or microbiostatic activity against selected bacteria, fungi,
and algae in vitro. Can. J. Microbiol., 48, 262-267.
Krieg, A. and Langenbruch, G.A. (1981) Susceptibility of arthropod species to
Bacillus thuringiensis. In Burges, H.D. (ed.) Microbial control of pests
and plant diseases 1970-1980, pp. 837-896.
Page 29 of 33
MON 810 maize
May 2007
Lang, A., Ludy, C. and Vojtech, E. (2004) Dispersion and deposition of Bt maize
pollen in field margins. Zeitschrift für Pflanzenkrankheiten und
Pflanzenschutz (J. Plant Diseases and Protection), 111, 417-428.
Lovei, G.L. and Arpaia, S. (2005) The impact of transgenic plants on natural
enemies: a critical review of laboratory studies. Entomologia
Experimentalis et Applicata, 114, 1-14.
Lozzia, G., Furlanis, C., Manachini, B. and Rigamonti, L. (1998) Effects of Bt
corn on Rhopalosiphum padi L. (Rhynchota Aphididae) and on its
predator Chrysoperla carnea Stephen (Neuroptera Chrysopidae). Boll.
Zool. Agraria Bachicol., 30, 153-164.
Ludy, C. and Lang, A. (2006a) A 3-year field-scale monitoring of foliage dwelling
spiders (Araneae) in transgenic Bt maize fields and adjacent field
margins. Biological Control, 38.
Ludy, C. and Lang, A. (2006b) Bt maize pollen exposure and impact on the
garden spider, Araneus diadematus. Entomologia Experimentalis et
Applicata, 118, 145-156.
MacIntosh, S.C., Stone, T.B., Sims, S.R., Hunst, P.L., Greenplate, J.T., Marrone,
P.G., Perlak, F.J., Fischhoff, D.A. and Fuchs, R.L. (1990) Specificity and
efficacy of purified Bacillus thuringiensis proteins against agronomically
important insects. J. Invertebr. Pathol., 56, 258-266.
Masoero, F., Moschini, M., Rossi, F., Prandini, A. and Pietri, A. (1999) Nutritive
value, mycotoxin contamination and in vitro rumen fermentation of
normal and genetically modified corn (Cry1A(B)) grown in northern
Italy. Maydica, 44, 205-209.
Meissle, M., Vojtech, E. and Poppy, G.M. (2005) Effects of Bt maize-fed prey on
the generalist predator Poecilus cupreus L. (Coleoptera: Carabidae).
Transgenic Research, 14, 123-132.
Melin, B.E. and Cozzi, E.M. (1990) Safety to nontarget invertebrates of
lepidopteran strains of Bacillus thuringiensis and their Beta exotoxins.
Safety of microbial insecticides, 149-167.
Mendelsohn, M., Kough, J., Vaituzis, Z. and Matthews, K. (2003) Are Bt crops
safe? Nature Biotechnology, 21, 1003-1009.
Monsanto Company. (1995) Submission to the French Commission du Génie
Biomoléculaire. Application to place on the market genetically modified
higher plants: insect-protected maize (MON810). Monsanto report.
Page 30 of 33
MON 810 maize
May 2007
Munkvold, G.P., Hellmich, R.L. and Rice, L.G. (1999) Comparison of fumonisin
concentrations in kernels of transgenic Bt maize hybrids and nontransgenic hybrids. Plant disease, 83, 130-138.
Naranjo, S., Head, G. and Dively, G. (2005) Field studies assessing arthropod
non-target effects in Bt transgenic crops. Environ. Entomol., 34, 11781180.
Naranjo, S.E. (2005a) Long-term assessment of the effects of transgenic Bt
cotton on the abundance of nontarget arthropod natural enemies.
Naranjo, S.E. (2005b) Long-term assessment of the effects of transgenic Bt
cotton on the function of the natural enemy community. Environm.
Entomol., 34, 1211-1223.
Obrist, L.B., Dutton, A., Albajes, R. and Bigler, F. (2006) Exposure of arthropod
predators to Cry1Ab toxin in Bt maize fields. Ecological Entomology, 31,
143-154.
Orr, D.R. and Landis, D.A. (1997) Oviposition of European corn borer
(Lepidoptera: Pyralidae) and impact of natural enemy populations in
transgenic versus isogenic corn. J. Econ. Entomol., 90, 905-909.
Palm, C.J., Donegan, K., Harris, D. and Seidler, R.J. (1994) Quantification in
soil of Bacillus thuringiensis var. kurstaki delta-endotoxin from
transgenic plants. Molecular Ecology, 3, 145-151.
Pilcher, C.D., Obrycki, J.J., Rice, M.E. and Lewis, L.C. (1997) Preimaginal
development, survival and field abundance of insect predators on
transgenic Bacillus thuringiensis Corn. Biological Control, 26, 446-454.
Pilcher, C.D., Rice, M.E. and Obrycki, J.J. (2005) Impact of transgenic Bacillus
thuringiensis corn and crop phenology on five nontarget arthropods.
Pleasants, J.M., Hellmich, R.L., Dively, G.P., Sears, M.K., Stanley-Horn, D.E.,
Mattila, H.R., Foster, J.E., Clark, T.L. and Jones, G.D. (2001) Corn
pollen deposition on milkweeds in and near cornfields. Proc. Natl. Acad.
Sci. USA, 98, 11919-11924.
Pruett, C.J.H., Burges, H.D. and Wyborn, C.H. (1980) Effect of exposure to soil
on potency and spore viability of Bacillus thuringiensis. J. Invertebr.
Pathol., 35, 168-174.
Page 31 of 33
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May 2007
Raps, A., Kehr, J., Gugerli, P., Moar, W.J., Bigler, F. and Hilbeck, A. (2001)
Immunological analysis of phloem sap of Bacillus thuringiensis corn and
of the non-target herbivore Rhopalosiphum padi (Homoptera: Aphididae)
for the presence of Cry1Ab. Molecular Ecology, 10, 525-533.
Reed, G.L., Jensen, A.S., Riebe, J., Head, G. and Duan, J.J. (2001) Transgenic Bt
potato and conventional insecticides for Colorado potato beetle
management: comparative efficacy and non-target impacts. Entomologia
Rice, M.E. and Pilcher, C.D. (1999) Bt corn and insect resistance management:
farmer perceptions and educational opportunities. A poster presented at
the 1999 meeting of the Entomological Society of America.
Romeis, J., Bartsch, D., Bigler, F., Candolfi, M., Gielkens, M., Hartley, S.,
Hellmich, R., Huesing, J., Jepson, P., Layton, R., Quemada, H.,
Raybould, A., Rose, R., Schiemann, J., Sears, M., Shelton, A., Sweet, J.,
Vaituzis, Z. and Wolt, J. (2006a) Moving through the tiered and
methodological framework for non-target arthropod risk assessment of
transgenic insecticidal crops. Proceedings of the 9th International
Symposium on the Biosafety of Genetically Modified Organisms, 62-67.
Romeis, J., Dutton, A. and Bigler, F. (2004) Bacillus thuringiensis toxin
(Cry1Ab) has no direct effect on larvae of the green lacewing Chrysoperla
carnea. Journal of Insect Physiology, 50, 175-183.
Romeis, J., Meissle, M. and Bigler, F. (2006b) Transgenic crops expressing
Bacillus thuringiensis toxins and biological control. Nature
Biotechnology, 24, 63-71.
Saxena, D. and Stotzky, G. (2001) Bacillus thuringiensis (Bt) toxin released from
root exudates and biomass of Bt corn has no apparent effect on
earthworms, nematodes, protozoa, bacteria, and fungi in soil. Soil Biol.
and Biochem., 33, 1225-1230.
Sims, S.R. and Holden, L.R. (1996) Insect bioassay for determining soil
degradation of Bacillus thuringiensis subsp. kurstaki CryIA(b) protein in
corn tissue. Environmental entomology, 25, 659-664.
Tiedje, J.M., Asuming-Brempong, S., Nusslein, K., Marsh, T.L. and Flynn, S.J.
(1999) Opening the black box of soil microbial diversity. Appl. Soil Ecol.,
13, 109-122.
Page 32 of 33
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May 2007
Tinland, B., Janssens, J., Lecoq, E., Legris, G., Matzk, A., Pleysier, A., Wandelt,
C. and Willekens, H. (2006) Implementation of general surveillance in
Europe: the industry perspective. J. Verbr. Lebensm., 1, 42-44.
Torres, J.B. and Ruberson, J.R. (2005) Canopy- and ground-dwelling predatory
arthropods in commercial Bt and non-Bt cotton fields: patterns and
mechanisms. Environ. Entomol., 34, 1242-1256.
Toth, F., Arpas, K., Szekeres, D., Kadar, F., Szentkiralyi, Szenasi, A. and Kiss, J.
(2004) Spider web survey or whole plant visual sampling? Impact
assessment of Bt corn on non-target predatory insects with two
concurrent methods. Environ. Biosafety Res., 3, 225-231.
US EPA. (2000) Bt-plant pesticides biopesticides registration document: section
C. Environmental Assessment. United States Environmental Protection
Agency.
US EPA. (2002) Memorandum. Transmittal of meeting minutes of the FIFRA
Scientific Advisory meeting held August 27-29, 2002. SAP meeting
minutes No. 2002-05. .
US
EPA. (2005) Biopesticides Registration Action Document. Bacillus
thuringiensis Cry34Ab1 and Cry35Ab1 proteins and the genetic material
necessary for their production (plasmid insert PHP 17662) in event DAS59122-7 corn. , 42.
Van Rie, J., Jansens, S., Hofte, H., Degheele, D. and Van Mellaert, H. (1989)
Specificity of Bacillus thuringiensis delta-endotoxins-importance of
specific receptors on thebrush border membrane of the mid-gut of target
insect. Eur. J. Biochem., 186, 239-247.
Receptors on the brush border membrane of the insect midgut as
determinants of the specificity of Bacillus thuringiensis Deltaendotoxins. Applied and environmental microbiology, 1378-1385.
Vercesi, M.L., Krogh, P.H. and Holmstrup, M. (2006) Can Bacillus thuringiensis
(Bt) corn residues and Bt-corn plants affect life-history traits in the
earthworm Aporrectodea caliginosa? Applied Soil Ecology, 32, 180-187.
Vinson, S.B. (1989) Potential impact of microbial insecticides on beneficial
arthropods in the terrestrial environment. Safety of Microbial
Insecticides, 43-64.
Page 33 of 33
MON 810 maize
May 2007
Vojtech, E., Meissle, M. and Poppy, G.M. (2005) Effects of Bt Maize on the
herbivore Spodoptera littoralis (Lepidoptera: Noctuidae) and the
parasitoid
Cotesia
marginiventris
(Hymenoptera:
Braconidae).
Volkmar, C. and Freier, B. (2003) Spinnenzoenosen in Bt-mais un nicht
gentechnisch
veränderten
Maisfeldern.
Zeitschrift
für
Pflanzenkrankheiten und Pflanzenschutz (J. Plant Diseases and
Protection), 110, 572-582.
Wandeler, H., Bahylova, J. and Nentwig, W. (2002) Consumption of two Bt and
six non-Bt corn varieties by the woodlouse Porcellio scaber. Basic and
Applied Ecology, 3, 357-365.
West, A.W. (1984) Fate of the insecticidal, proteinaceous parasporal crystal of
Bacillus thuringiensis in soil. Soil Biol. Biochem, 16, 357-360.
West, A.W., Burges, H.D., White, R.J. and Wyborn, C.H. (1984) Persistence of
Bacillus thuringiensis parasporal crystal insecticidal activity in soil. J.
Invertebr. Pathol., 44, 128-133.
Whitehouse, M., Wilson, L. and Fitt, G. (2005) A comparison of arthropod
communities in transgenic Bt and conventional cotton in Australia.
Wilhelm, R., Beissner, L. and Schiermann, J. (2003) Concept for the realisation
of a GMO monitoring in Germany. Federal Biological Research Centre for
Agriculture and Forestry, Institute for Plant Virology, Microbiology and
Biosafety.
Wolfersberger, M.G., Hofmann, C. and Luthy, P. (1986) Interaction of Bacillus
thuringiensis delta-endotoxin with membrane versicles insolated from
lepidoteran larval midgut. Bacterial protein toxins, 237-238.
Wolt, J., Conlan, C. and Majima, K. (2005) An ecological risk assessment of
Cry1F maize pollen impact to pale grass blue butterfly. Environmental
Biosafety Research, 4, 243-251.
Zwahlen, C. and Andow, D.A. (2005) Field evidence for the exposure of ground
beetles to Cry1Ab from transgenic corn. Environ. Biosafety Res, 4, 113117.
Zwahlen, C., Hilbeck, A., Gugerli, P. and Nentwig, W. (2003) Degradation of the
Cry1Ab protein within transgenic Bacillus thuringiensis corn tissue in
the field. Molecular Ecology, 12, 765-775.
MON 810 maize
May 2007
APPENDIX 1
Environmental Risk Assessment from
Application for renewal of the
authorisation for continued marketing
of existing MON 810 maize products that
were authorized under Directive
90/220/EEC (Decision 98/294/EC) and
subsequently notified in accordance to
Article 20(1)(a) of Regulation (EC)
No 1829/2003 on genetically modified food
and feed
Part I
Technical Dossier
May 2007
Data protection.
This application contains scientific data and other information which are protected in accordance with Art. 31 of
Regulation (EC) No 1829/2003.
 2007 Monsanto Company. All Rights Reserved.
This document is protected under copyright law. This document is for use only by the regulatory authority to which this
has been submitted by Monsanto Company, and only in support of actions requested by Monsanto Company. Any other
use of this material, without prior written consent of Monsanto, is strictly prohibited. By submitting this document,
Monsanto does not grant any party or entity any right to license, or to use the information of intellectual property
described in this document.
Part I – Technical dossier
1
Regulation (EC) No 1829/2003
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TABLE OF CONTENTS
Page
9. Potential changes in the interactions of the GM plant with the
biotic environment resulting from the genetic modification...................... 4
9.1
Persistence and invasiveness .................................................. 4
9.2
Selective advantage or disadvantage ...................................... 7
9.3
Potential for gene transfer..................................................... 10
9.4
Interactions between the GM plant and target organisms .. 13
9.5
Interactions of the GM plant with non-target organisms .... 15
9.6
Effects on human health........................................................ 26
9.7
Effects on animal health........................................................ 28
9.8
Effects on biogeochemical processes...................................... 31
9.9
Impact of the specific cultivation, management and
harvesting techniques............................................................ 34
References........................................................................................................ 38
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LIST OF TABLES
Table 1.
Summary of Monsanto’s laboratory and greenhouse studies
investigating the potential adverse effects of Cry1Ab protein
and lepidopteran-protected crops containing Cry1Ab protein
on pest and beneficial organisms compared to conventional
varieties...............................................................................................19
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9.
Potential changes in the interactions of the GM plant with the
biotic environment resulting from the genetic modification
As this renewal application under Regulation (EC) No 1829/2003 includes
the cultivation of MON 810 in the E.U., a complete environmental risk
assessment was conducted according to Annex II of Directive 2001/18/EC
on the deliberate release of genetically modified organisms in the
environment. Nine potential adverse environmental effects are laid out in
Annex II of Directive 2001/18/EC, which could occur at least in theory
when a GM higher plant (GMHP) is placed on the market in Europe. The
potential for MON 810 to adversely impact the environment in the E.U. is
evaluated below, according to the recommended step-wise risk
assessment process, including: a) identification of potentially harmful
characteristics of the GMHP; b) potential consequence of the theoretical
adverse effect (assuming fully realised); c) likelihood of the theoretical
adverse effect to occur for this product and a characterisation of the
actual hazard potential of the identified GMHP characteristic;
d) estimation of the risk and e) necessary risk mitigation measures, if
applicable.
In the previous sections of this application, MON 810 was shown not to be
different from conventional maize in its agronomic, phenotypic,
compositional, nutritional and safety characteristics, suggesting that any
interactions of this maize with the biotic environment have not been
changed compared to conventional maize. No adverse environmental
effects are to be expected for MON 810.
9.1 Persistence and invasiveness
Equivalent data requirement according to Annex II to Directive
2001/18/EC: “Likelihood of the GMHP becoming more persistent
than the recipient or parental plants in agricultural habitats or more
invasive in natural habitats.”
a) Characteristics of the GMHP, which may cause an adverse
effect
Based on centuries of experience with conventional,
domesticated maize in Europe, there is no potential for maize to
be invasive of natural habitats or persist in or outside the
agricultural environment without the aid of human intervention.
Maize is a poor competitor of plants, which outside of cultivation
has no meaningful impact on biodiversity or the environment.
Extensive characterization of MON 810 (which, in part, includes
molecular, expression, composition, and phenotypic data)
demonstrated that the only meaningful difference between
MON 810 and conventional maize is the lepidopteran protection
trait conferred by the Cry1Ab protein. This is evaluated in this
section.
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b) Potential consequences of the adverse effect, if it occurs
At full magnitude of the consequences thereof, increased
persistence could, in part, elevate maize to weed status and
could result in an invasive species, spreading in the
environment. However, such changes or impact would be
atypical of the plant Zea mays, and have not been reporterd
through the decades of plant breeding, mutagenesis and other
forms of introducing genetic diversity into maize.
c) Likelihood of the occurrence of the potential adverse effect
Comparative assessment of weediness potential
This renewal application is for the placing on the market of
MON 810 in the E.U. The proposed uses of this maize are the
same as for any other maize, including the cultivation of varieties
in the field. MON 810 may be cultivated or used in any
environment currently suitable for the production or use of
maize. Therefore, the environmental release conditions of
MON 810 would not be different from those for any other maize.
Conventional maize, originally introduced into Europe over 500
years ago, is an annual crop that is not inherently persistent or
invasive. It cannot survive without human assistance and is not
capable of surviving as a weed due to centuries of breeding and
selection. Despite some 6 million hectares of maize being grown
in the E.U.-25 annually (FAOSTAT1), and the resulting harvest
transported many thousands of kilometres by road, railway and
waterways, persistent populations of maize are not found growing
in fencerows, ditches, and roadsides, nor in natural habitats
further removed from agriculture. Surveys of spontaneous plant
populations in set-aside fields in France (Bodet et al., 1994;
Mamarot and Rodriguez, 1994) have documented that selfsustaining populations of maize are not present. This results from
the combination of the absence of seed dormancy, the poor
survivability of seed in soils, the frost sensitivity of maize
seedlings and the soil preparations prior to the planting of
subsequent crops (which includes destruction of any existing
vegetation and soil cultivation) (Hicks and Thomison, 2004;
OECD, 2003; Shaw, 1988).
Seed kernels are the only survival structures of maize; natural
regeneration from vegetative tissue is not known to occur. In
contrast to many weedy plants, maize has a polystichous female
inflorescence (ear) on a stiff central spike (cob) enclosed in husks
(modified leaves). This structure does not pre-dispose the
individual kernels to natural dissemination. Nonetheless, in a
1
http:// faostat.fao.org/site/408/default.aspx
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cultivation scenario, kernels may be disseminated by mechanical
harvesting, by insect or wind damage and by wild animals
foraging on the crop, all of which may cause some mature ears or
kernels to fall to the ground, where they could remain after
harvest. Although these kernels can over-winter under mild
conditions and can germinate the following year, maize cannot
persist as a weed (Hallauer, 1995; OECD, 2003). The appearance
of maize volunteer plants developing from shed kernels or the
appearance of maize in rotational fields following the maize crop
from the previous year are rare under most European conditions
as maize volunteers are killed by frost or easily controlled by
current agronomic practices, including cultivation or the use of
selective herbicides in the next crop.
The above observations are not different for MON 810. As
established in Section D.7 of this renewal application, MON 810
is not substantially different from conventional maize, except for
the introduced lepidopteran-protection trait. Field trial data for
MON 810 have demonstrated that this maize has not been
altered in its phenotypic, agronomic, reproductive, survival and
dissemination characteristics when compared to conventional
maize. This is supported by positive experience from commercial
planting of MON 810 in the E.U., North America and elsewhere.
Given that the genetic modification did not alter the phenotypic
characteristics of this maize compared to conventional
counterparts, it is highly unlikely that MON 810 would be any
more persistent in its receiving environment or more invasive in
non-agricultural environments than conventional maize.
In conclusion, it is highly unlikely that MON 810 would be
weedier compared to conventional maize. Therefore, the
likelihood of this maize to spread beyond the agricultural
environment where it is grown is negligible.
Evaluation of the potential of the GMHP to cause adverse effects, if
the plant would establish
The introduced lepidopteran-protection trait confers a selective
advantage only under specific conditions (i.e. upon attack by the
target insects), which are short in duration. The advantage is of
agronomic interest and presents negligible risk to nonagricultural environments, because of the poor survival
characteristics of maize under most European conditions. This is
discussed in detail in Section D.9.2.
Similarly, in the unlikely case where kernels would germinate
outside of the field, the lepidopteran-protection trait would not
confer any meaningful advantage to the emerging plant, as maize
is incapable of surviving without human assistance under
European conditions. In the field, this trait would only confer a
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competitive advantage over untreated conventional maize under
the specific condition of herbivorous attack on the crop by one of
the susceptible lepidopteran insect pests (i.e. the desired
agronomic lepidopteran-protection trait in MON 810). However,
this selective advantage of one maize over another in the field is
not relevant to natural ecosystems.
d) Estimation of the risk
It is concluded that, like for conventional maize, the likelihood of
MON 810 adversely impacting the environment is negligible, as it
has shown no ability to be persistent or invasive and these
parameters are unaltered in MON 810 when compared to
conventional maize. In the unlikely event of the establishment of
MON 810 plant in the environment, the introduced trait would
confer only a limited selective advantage (protection from
lepidopteran pests) of short duration, narrow spatial context and
have negligible consequences for the environment. Hence the risk
to the environment from MON 810 through increased
persistence and invasiveness of this maize is negligible.
e) Application of risk management strategies
As the risk is negligible, risk management strategies are not
considered necessary.
Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Presence of the
introduced
lepidopteranprotection trait
Increased
persistence or
invasiveness,
resulting in an
invasive species
spreading in the
environment
Negligible
Negligible
(not applicable)
9.2
Selective advantage or disadvantage
2001/18/EC: “Any selective advantage or disadvantage conferred to
the GMHP”
effect
Extensive characterization of MON 810 (which, in part, includes
molecular, expression, composition, and phenotypic data)
demonstrated that the only meaningful difference between
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MON 810 and conventional maize is the lepidopteran protection
trait conferred by the Cry1Ab protein. Therefore, the introduced
trait is a characteristic of the GMHP that may, at least in
theory, cause an adverse environmental effect. This is evaluated
in this section.
If the lepidopteran protection trait was to confer a selective
advantage, then at full magnitude of the consequences thereof,
this could result in a maize plant that could out-compete native
vegetation
and
potentially
invade
non-agricultural
environments.
Presence of the new trait in the grain
Compared to conventional maize, the newly introduced trait in
MON 810 is limited to the expression of Cry1Ab protein
conferring the lepidopteran-protection. It was demonstrated
previously that the introduced genetic sequence in MON 810 did
not lead to altered phenotypic characteristics, such as plant
growth and development, morphology, agronomic performance,
composition, or nutritional value, when compared to
conventional maize. It was concluded that MON 810 is not
substantially different from conventional maize, with the
exception of the intentionally introduced lepidopteran-protection
trait.
As no other new traits were introduced, the assessment of any
competitive (dis)advantages in the following paragraph will be
limited to the lepidopteran-protection trait.
No meaningful competitive (dis)advantage conferred by the
introduced trait
Compared with conventional maize, the presence of the
lepidopteran-protection trait would only confer a selective
advantage where target lepidopteran pest species including
European corn borer (Ostrinia nubilalis) and pink borers
(Sesamia spp.), would be present at sufficiently high numbers to
limit reproductive success, and if no other, more important
factors limiting the survival of maize in the receiving
environment would be present. The expression of this protein
does not confer a selective advantage or disadvantage to this
maize in the natural environment, as in any case, MON 810 is
not a weedy plant and for the reasons described in Section
D.9.3., the likelihood is negligible for MON 810 to volunteer or
survive in natural habitats under most European climatic
conditions (see Section D.9.1).
8
MON 810
Monsanto Company
Strictly taken, the introduced trait in MON 810 plants has a
competitive advantage to maize in agricultural habitats. The
lepidopteran-protection trait provides a selective advantage
to MON 810 over (untreated) conventional maize plants in the
field when there is actual herbivorous pressure from susceptible,
target insect pests.
This introduced “competitive advantage” is only relevant in
agricultural habitats (i.e. in the corn fields) and is limited in time
from late emergence until harvest, when lepidopteran pest
pressure
is
highest,
because
any
eventually
non-harvested MON 810 plants would not likely persist in the
field (see Section D.9.1). Moreover, rare volunteers are readily
controlled by mechanical means, or by one of a number of the
other graminicides currently used. Finally, these “advantages” of
lepidopteran-protected MON 810 over unprotected conventional
maize plants in the agricultural habitat are not ecologically
meaningful when they are viewed in the context of today’s
baseline agronomic practices for the production of maize. One
most commonly adopted management technique in conventional
agriculture, in order to maximise crop yield and harvest quality,
is to control insect pests with insecticides, thereby causing lethal
or otherwise very disruptive effects on target insects in the crop.
Therefore, the likelihood is negligible for the introduced trait
in MON 810 to confer any meaningful competitive advantage or
disadvantage of relevance to the environment.
The introduced lepidopteran-protection trait confers a selective
advantage only under specific conditions (i.e. upon attack by the
target insects), which are short in duration. The advantage is of
purely agronomic interest and presents negligible risk to the
non-agricultural environments, because of the poor survival
characteristics of maize under most European conditions.
In conclusion, the risk of the introduced trait in MON 810 to be
the cause of any adverse effects resulting from a competitive
advantage or disadvantage in natural environments is
negligible.
Within commercial MON 810 fields, the maize plants
theoretically have a selective advantage over unprotected
conventional maize plants under specific conditions in the field
(i.e. under actual herbivory by target insect pests). However,
these conditions are predictable, spatially limited, short in
duration and with negligible consequences to the environment.
These “selective advantages” are limited to the agricultural field
9
MON 810
Monsanto Company
and the growing season of the MON 810 crop, and are considered
of negligible risk to the agronomic and natural environment.
As the risk is negligible, no risk management strategies are
considered.
Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Presence of the
introduced
Competitive
advantage of
MON 810,
resulting in an
invasive species
spreading in the
environment
Negligible
Negligible
(not applicable)
9.3
Potential for gene transfer
2001/18/EC: “Potential for gene transfer to the same or other
sexually compatible plant species under conditions of planting the
GMHP and any selective advantage or disadvantage conferred to
those plant species.”
effect
All maize produced in Europe can inter-pollinate. MON 810, like
all other maize, is not sexually compatible with any indigenous or
introduced wild plant species present in Europe. Therefore, the
potential for genetic transfer and exchange with other plants is
limited to cross-pollination to cultivated maize plants. MON 810
is unchanged in its potential for gene transfer compared to
conventional maize.
Maize pollen of a specific cultivar can be carried over short
distances by the wind and could fertilise other cultivars. In the
event of plants of MON 810 maturing, producing pollen and
fertilising a neighbouring crop, the introduced lepidopteranprotection trait present in MON 810 could be transferred to a
recipient maize crop, and could be expressed in the progeny of
the recipient crop. Potential outcrossing of the introduced trait
is a characteristic of the GMHP that may, theoretically, cause an
adverse environmental effect.
10
MON 810
Monsanto Company
Outcrossing itself is not an adverse event. If outcrossing of an
introduced trait to sexually compatible plants would occur and
conferred an advantage, then at the full magnitude of the
consequences thereof, this could result in a maize plant that
could have more offspring that survives outside of cultivation
and potentially be fitter than conventional maize in unmanaged
environments. However, if the genetic modification were to
confer a disadvantage, this could not result in maize plants that
produce less offspring and are less competitive compared to other
maize. However, as established in Section D.9.2 for the source
crop itself, the risk of the lepidopteran-protection trait to be the
cause of any meaningful competitive advantage or disadvantage
that could impact the receiving environment is negligible.
Therefore, no adverse effect would occur.
Incidence of out-crossing of the introduced trait
The potential for transfer of genetic material between maize crops
is limited by the mobility of maize pollen. Measuring about 0.1 mm
in diameter, maize pollen is the largest of any pollen normally
disseminated by wind from a comparably low level of elevation.
Due to its relatively large mass, the majority of maize pollen
does not move more than a few meters from the crop. Most maize
pollen falls within five meters of the field edge (Pleasants et al.,
2001; Sears et al., 2001). Hansen (1999) found that in adjacent
leaves to a B.t. maize field at 0, 1 and 3 m distance from it,
pollen deposition decreased significantly. The Sears et al. (2001)
study showed that a cumulative 99% of maize pollen was
measured at 50 m and 100% at 100 m from the source crop. In a
study of out-crossing with maize undertaken in France (AGPM,
1999), pollen flow as measured by successful fertilisation of
neighbouring maize declined to 1% at a distance of 10 m from
the source crop. Other research confirmed that, on average, most
maize pollen travels no further than 100 metres although a cutoff distance is not clear (Devos et al., 2005) and that crosspollination is barely detectable at 200 m (<0.1%) (Halsey et al.,
2005) and nonexistant at 300 m (Luna et al., 2001). Nearly all
potential cross-pollination between maize fields occurs within 30
metres of the pollen source (Ma et al., 2004; Messeguer, 2003).
The likelihood of outcrossing of traits between maize plants
further depends on their synchrony of flowering, the volume of
pollen produced, the distance between the crops, size of the fields,
physical barriers, e.g., trees, and their orientation on the
11
MON 810
Monsanto Company
landscape, i.e. down-wind vs. up-wind during pollination (Devos
et al., 2005).
No adverse effects associated with outcrossing of the introduced
trait
As established for the MON 810 source crop itself, outcrossing of
the introduced trait to the neighbouring maize crop would confer
a selective advantage only upon conditions where it is infested by
target insect pests. Any advantage would be predictable,
spatially limited, short in duration, and have no adverse
consequences for the agricultural or natural environment (see
Section D.9.2.). The potential for any resulting F2 grain,
containing the new genetic sequences, to germinate and survive
as volunteer maize plants or to spread to the environment is
negligible, as discussed in Section D.9.1 for the source crop.
In conclusion, there is no potential for gene transfer from
MON 810 to wild plant species in the E.U. There is a significant
likelihood for gene transfer to other maize crops, depending on
wind, flowering synchrony and distance between the crops. In the
event that an introduced gene would outcross to other maize, its
transfer would not confer a selective advantage as discussed in
Section D.9.2. Therefore, it is not considered to constitute an
adverse environmental effect in itself. The environmental risk
posed by this transfer, and hence by MON 810 is negligible.
e) Application of management strategies
• Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence
of the adverse
effect, if it
occurs
• Sexual
compatibility
with other maize,
allowing crosspollination
Transfer of any
selective
advantage to
other maize
plants, which
could become
invasive and
spread in the
environment
• Presence of the
introduced
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Negligible
Negligible
(not applicable)
12
MON 810
Monsanto Company
9.4
Interactions between the GM plant and target organisms
Equivalent data requirement according to Annex II of Directive
2001/18/EC: “Potential immediate and/or delayed environmental
impact resulting from direct and indirect interactions between the
GMHP and target organisms (if applicable).”
effect
The insecticidal action of the Cry1Ab protein to target
lepidopteran pests including the European corn borer (Ostrinia
nubilalis) and pink borers (Sesamia spp.) has been identified as
the characteristic, which may cause an adverse effect. Control of
pest species is not considered adverse to the agricultural
environment. However, the expression of newly introduced
protein in a GMHP could, at least in theory, cause adverse
environmental effects through direct or indirect interactions
between the GMHP and target organisms (where applicable).
The potential for adverse environmental effects resulting from
interactions of this maize with these target organisms is
evaluated in this section.
MON 810 has been developed to provide control against certain
lepidpteran pests of maize. The success of MON 810
commercially is dependent on maintaining sufficient levels of
expression of Cry1Ab throughout the season when lepidopteran
pests are present, and delaying the development of resistance to
the target organisms. Fields of MON 810 in the E.U. are expected
to have low abundance of the target organisms, which will have
an adverse effect on specialist predators and parasitoids.
However, threatened or endangered specialist predators and
parasitoids of the target organisms are unknown in the E.U.
Furthermore, the target pests are known to infest several other
weedy non-agricultural plants thus providing hosts and prey for
predators and parasitoids outside of the agricultrual
environment. Since predators and parasitoid populations would
be adversely impacted by any other lepidopteran pest control
measures, MON 810 poses no increased risk to these organisms.
Consequently, the only identified potential consequence of an
adverse effect, if it occurs, would be the development of
resistance in the target pests to the insecticidal Cry1Ab protein
expressed in MON 810.
13
MON 810
Monsanto Company
The mechanism of action of Bt Cry proteins including Cry1Ab in
susceptible insects is well-known. Bt Cry proteins bind to unique
receptor sites on the insect midgut epithelial membrane causing
development of pores, disruption of osmotic balance, and
ultimately septicemia (Broderick et al., 2006; Gill and Ellar,
2002). Each class of Cry proteins, e.g., Cry1A proteins, has been
shown to be highly specific for select orders of insects. In the
case of the Cry1A proteins it has been established that these
proteins exhibit selective toxicity towards certain lepidopteran
pests, but not against other insect orders at levels found in
MON 810. Within the E.U., the insecticidal action of the Cry1Ab
protein is to target lepidopteran pests including the European
corn borer (Ostrinia nubilalis) and pink stem borers (Sesamia
spp.).
Receptor binding, in particular, is a critical step in the
mechanism of action of Cry proteins because, without it, no toxic
effect can be exerted. Irreversible binding of toxins to midgut
receptors appears to be correlated with insect susceptibility. The
Cry1A proteins bind specifically to receptors on the midgut of
lepidopteran insects (Hofmann et al., 1988a; Hofmann et al.,
1988b; Van Rie et al., 1989; Van Rie et al., 1990; Wolfersberger et
al., 1986).
Resistance evolution in targeted lepidopteran pests is a potential
concern arising from the widespread cultivation of MON 810.
However, in those countries where MON 810 has been planted,
insect resistance management (IRM) plans have been put in
place to minimize the risk of insect resistance evolving to
Cry1Ab. This will continue to be the case wherever MON 810 is
grown. These IRM plans routinely include setting aside refuges
for the production of susceptible target insects, educating
farmers as to the importance of IRM, measuring the
susceptibility of target insects prior to widespread product use,
and putting in place insect resistance monitoring programs. For
example, prior to MON 810 being grown in Spain, baseline
susceptibility studies were conducted for both of the primary
target pest species (Gonzalez-Nunez et al., 2000), and resistance
monitoring has been conducted subsequently. In the U.S.,
Argentina, and other countries, extensive cultivation of
MON 810 has occurred since 1996 in conjunction with approved
IRM plans without any resistance evolving to MON 810
(Tabashnik et al., 2003). Therefore, the risk of resistance evolving
to the Cry1Ab protein in target organisms through the use of
MON 810 is minimal.
14
MON 810
Monsanto Company
Since IRM plans are put in place in those countries where
MON 810 is planted the potential for insect resistance to Cry1Ab
to occur will be negligible.
Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Protection from
lepidopteran
insect pests
conferred by the
expression of the
Cry1Ab protein
in the plant
Development of
resistance to
Cry1Ab. by certain
lepidopteran
species
Negligible
Negligible
IRM Plan in
cultivation
countries
9.5
Interactions of the GM plant with non-target organisms
2001/18/EC: “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 target organisms), including impact on
population levels of competitors, herbivores, symbionts (where
applicable), parasites and pathogens.”
effect
Like any other plant, cultivated maize is known to interact with
a variety of organisms in the environment, including
microorganisms, wildlife and numerous soil dwelling and foliar
dwelling invertebrates. In addition, maize is known to be
susceptible to a range of fungal diseases and nematodes, insect
and mite pests, which the grower traditionally has attempted to
control by the application of plant protection products or by
means of other agricultural practices such as crop rotation.
Because maize is a good source of nutrition, interactions with
vertebrate wildlife are well-known, including with birds and
mammals that reside or forage in the agricultural habitat made
up by the crop and its field edges, hedgerows or ditches. As
MON 810 was shown not to be substantially different from
conventional maize (except for the introduced lepidopteran
protection), its baseline interaction with other organisms in the
15
MON 810
Monsanto Company
environment is considered no different than conventional maize,
except for the additional potential exposure of herbivorous pests
of maize and grazing animals to the Cry1Ab protein that is
newly expressed in the plant. Through trophic transfer and
decomposition processes, additional organisms such as predators
could be exposed to this protein expressed in maize in some
diluted manner. Potential exposure of non-target organisms in
the receiving environment to this protein is a characteristic of
the GMHP that may, theoretically, cause an adverse
environmental effect.
Non-target organisms include all organisms, animals and plants,
which may unintentionally be affected through a specific or nonspecific mechanism, as a result of the newly expressed Cry1Ab
protein. Theoretically, potential toxicity of the newly expressed
protein to non-target organisms could affect their population
levels in the receiving environment, which is evaluated in this
section. As the Cry1Ab is toxic to certain (targeted) lepidopteran
insects, assessment of potential activity of this protein on nontarget organisms is central to this evaluation.
c)
Likelihood of the occurrence of the potential adverse effect
The likelihood for adverse environmental effects resulting from
the exposure of non-target organisms to the newly expressed
Cry1Ab protein in MON 810 is negligible as the safety
assessment for MON 810 indicated negligible environmental
risks.
MON 810 expresses the introduced Cry1Ab protein, protecting
the plants against herbivorous predation by important insect
pests of maize. It has been established that this Cry protein
exhibits selective toxicity towards certain lepidopteran pests, but
not against other insect orders. This specificity is consistent with
findings reported in the published literature, that demonstrate
that proteins of the Cry1A class exhibit lepidopteran selective
toxicity (Aronson and Shai, 2001; Crickmore et al., 1998;
Crickmore et al., 2005; Dulmage, 1981; Klausner, 1984;
MacIntosh et al., 1990; Mendelsohn et al., 2003; Romeis et al.,
2006; Whiteley and Schnepf, 1986).
The insecticidal specificity of Cry proteins can be determined by
any number of steps in the mode of action, previously described
in Section D.8. Receptor binding, in particular, is a critical step
in the mechanism of action of Cry proteins because, without it, no
toxic effect can be exerted. Irreversible binding of toxins to
midgut receptors appears to be correlated with insect
susceptibility. The Cry1A proteins bind specifically to receptors
16
MON 810
Monsanto Company
on the midgut of lepidopteran insects (Hofmann et al., 1988a;
Hofmann et al., 1988b; Van Rie et al., 1989; Van Rie et al., 1990;
Wolfersberger et al., 1986) and have no deleterious effect on
beneficial or other non-target insects, including predators and
parasitoids of lepidopteran insect pests or honeybees (Apis
mellifera) (Cantwell et al., 1972; Flexner et al., 1986; Krieg and
Langenbruch, 1981; Melin and Cozzi, 1990; US EPA, 2000;
Vinson, 1989). Selectivity based on the mode of action is a key
factor in the safety of Cry proteins for non-target organisms such
as fish, birds, mammals and non-target invertebrates. No
receptors for these proteins have been identified in the intestinal
cells of mammals to date (Emmerling et al., 2001; Garner et al.,
1999; Noteborn and Kuiper, 1994; Sacchi et al., 1986; Van
Mellaert et al., 1988).
The full length Cry1Ab protein encoded by the cry1Ab gene that
was used to produce lepidopteran-protected MON 810, and the
insecticidally active core protein produced in the insect gut
following ingestion, is identical to the respective full length and
trypsin-resistant core Cry1Ab proteins contained in microbial
formulations that have been used safely for nearly 40 years in
commercial Bt-sprays. There is extensive information on the
absence of non-target effects from the Cry1Ab protein
(Mendelsohn et al., 2003; Romeis et al., 2006).
To confirm and expand on the results produced for the microbial
products which contain the same Cry1Ab protein as produced in
MON 810, the potential impact of the Cry1Ab protein on nontarget organisms was assessed on several representative
organisms. These studies have previously been reported for
MON 810 (Monsanto Company, 1995) and are summarized in
Table 1. Studies were conducted with the trypsin-resistant core
of the Cry1Ab protein because this is the insecticidally active
portion of the Cry1Ab protein. Non-target species that were
tested include a) larval and adult honeybees (Apis mellifera L.),
which is a beneficial insect pollinator, b) green lacewing larvae
(Chrysopa carnea), a beneficial predatory insect; c) Hymenoptera
(Brachymeria intermedia), a beneficial parasite of the housefly;
d) the ladybird beetle (Hippodamia convergens), a beneficial
predaceous insect, and e) earthworms (Eisenia fetida), a
representative detritivorous species in the soil. In addition, leaf
material of MON 810 plants was used in a non-target soil
organism study using Collembola (Folsomia candida). Due to the
potential exposure of aquatic invertebrates to maize pollen
containing the Cry1Ab protein, a toxicity test was also performed
on daphnids (Daphnia magna). The U.S. EPA has since
determined that aquatic invertebrate tests are categorized as
supplemental studies since the likelihood of exposure of aquatic
17
MON 810
Monsanto Company
animals including invertebrates to maize pollen is low (US EPA,
2002; US EPA, 2005). The results of these non-target organism
studies showed that the mortality of non-lepidopteran insect
species and three other representative organisms exposed to the
Cry1Ab protein at levels in excess of potential environmental
exposures did not significantly differ from control mortality.
18
MON 810
Monsanto Company
Table 1.
Summary of Monsanto’s laboratory and greenhouse studies investigating the potential adverse effects of
Cry1Ab protein and lepidopteran-protected crops containing Cry1Ab protein on pest and beneficial
organisms compared to conventional varieties.
Test Substance
Tryptic B.t.k HD-11
Protein/ U.S.A.
Organism Tested
Green Lacewing
Larvae (Chrysoperla
(formerly Chrysopa)
carnea)
Tryptic B.t.k HD-1
Protein/U.S.A.
Parasitic
Hymenoptera
(Brachyrneria
intermedia)
Study Design
A gene encoding the full length B.t.k. HD-1 protein was
introduced into Escherichia coli, expressed, isolated and
converted to the trypsin-resistant core, and then purified,
characterized and assessed for potential effects on green
lacewing larvae (Chrysoperla carnea). The test system
consisted of a paste material coated onto the surface of
Sitotroga sp. moth eggs, which were subsequently fed to
lacewing larvae. Three paste treatments were tested: i)
trypsinized HD-1 protein at a nominal rate of 20 ppm, ii) heat
attenuated HD-1 protein control at a nominal rate of 20 ppm,
and iii) water only control. One larva was placed in each
treatment cup with 30 larvae in each treatment group. The
test was terminated on Day 7 when pupation in the water and
heat attenuated controls exceeded 20%.
characterized and assessed for potential effects on parasitic
Hymenoptera (Brachyrneria intermedia). The test system
consisted of one pint rolled paper test chambers covered at
both ends with a disposable plastic Petri dish. Each treatment
had two test chambers and each test chamber constituted a
replicate of 25 parasitic Hymenoptera each. Test diets were
prepared by mixing a calculated amount of the test material
with honey/water (50/50) syrup to achieve a nominal
concentration of 20 ppm B.t.k. Three treatments were tested:
i) trypsinized HD-1 protein at a nominal rate of 20 ppm, ii)
heat attenuated HD-1 protein control at a nominal rate of 20
ppm, and iii) water only control. Fresh diet was presented
every 3 days. Parasitic Hymenoptera were observed twice on
the day of test initiation for mortality and signs of toxicity
and once each day thereafter. The study was terminated after
30 days.
Conclusion
Green lacewing larvae exposed to activated B.t.k. HD-l
protein at a concentration of 20 ppm on moth eggs for
seven days did not exhibit treatment related mortality or
signs of toxicity.
Parasitic Hymenoptera exposed to activated B.t.k. HD-1
protein at a concentration of 20 ppm in a honey/water
solution for thirty days did not exhibit treatment related
mortality or signs of toxicity. The LC50 for B.t.k. HD-1
protein in parasitic Hymenoptera is greater than 20 ppm.
The no-observed effect level was 20 ppm.
19
MON 810
Monsanto Company
Reference
(Hoxter and Lynn,
1992a)
(Hoxter and Lynn,
1992c)
Table 1
organisms compared to conventional varieties. - continued
Test Substance
Tryptic B.t.k HD-1
Protein/U.S.A.
Organism Tested
Ladybird Beetle
(Hippodamia
convergens)
Tryptic B.t.k HD-1
Protein/U.S.A.
Honeybee larvae,
(Apis mellifera)
Study Design
introduced into E. coli, expressed, isolated and converted to
the trypsin-resistant core, and then purified, characterized
and assessed for potential effects on the ladybird beetle
(Hippodamia convergens). The test system consisted of one
pint rolled paper test chambers covered at both ends with a
disposable plastic Petri dish. Each treatment had two test
chambers and each test chamber constituted a replicate of 25
parasitic ladybird beetles each. Test diets were prepared by
mixing a calculated amount of the test material with
honey/water (50/50) syrup to achieve a nominal concentration
of 20 ppm B.t.k. Three treatments were tested: i) trypsinized
HD-1 protein at a nominal rate of 20 ppm, ii) heat attenuated
HD-1 protein control at a nominal rate of 20 ppm, and iii)
water only control. Fresh diet was presented every 3 days.
The test was terminated on day 9 when negative control
mortality exceeded 20%
introduced into E. coli, expressed, isolated and converted to
the trypsin-resistant core, and then purified, characterized
and assessed for potential effects on honeybee larvae (Apis
mellifera). The test system consisted of 1-4 day old larvae
within the larval cells of their brood frames. Each treatment
and control group had three replicates of 50 larval bees each.
Four treatments were tested: i) trypsinized HD-1 protein at a
nominal rate of 20 ppm, ii) heat attenuated HD-1 protein
control at a nominal rate of 20 ppm, iii) water only control
and iv) no treatment. Dosing was accomplished by placing 5.0
µL of a 1.0 mg/mL stock solution in distilled water or distilled
water alone into the wells with the larvae. After dosing,
treated frames were returned to the super for completion of
larval development. Adult bees emerging from the capped
cells were counted and placed into adult holding cages on a
daily basis. Percent larval survival (from dosing to adult
emergence) and post-emergent adult survival were compared
among the four treatments.
Conclusion
Ladybird beetles exposed to activated B.t.k. HD-1 protein
at a test concentration of 20 ppm in a honey/water solution
for 9 days did not exhibit treatment related mortality or
signs of toxicity. The LC50 for B.t.k. HD-1 protein in
ladybird beetle (Hippodamia convergens) is greater than
20 ppm. The no-observed effect level was 20 ppm.
Survival from capping through adult emergence was 100
percent in all treatments. Adult post-emergent survival
through trial termination ranged from 84 to 100 percent.
The LC50 for B.t.k. HD-1 protein in larval honey bees
(Apis mellifera) is greater than 20 ppm. The no observed
effect level was 20 ppm.
20
MON 810
Monsanto Company
Reference
(Hoxter and Lynn,
1992b)
(Maggi and Sims,
1994b)
Table 1
Test Substance
Tryptic B.t.k HD-1
Protein/U.S.A.
Organism Tested
Honeybee adult, (Apis
mellifera)
Tryptic B.t.k HD-1
Protein/U.S.A.
Earthworm (Eisenia
fetida)
Study Design
characterized and assessed for potential effects on honeybee
adults (Apis mellifera). The test system consisted of 3.2 mm
wire mesh metal hardware cloth containers with detachable
lids. Test material was introduced into the cage using a glass
shell vial fitted with a cotton wick. Adult honeybees were
obtained by randomly selecting young individuals from brood
frames located within bee hives. Three replicates with at
least 40 bees/container were used. Test diet consisted of a
50:50 (vol:vol) honey:water mixture. Three treatments were
tested: i) trypsinized HD-1 protein at a nominal rate of 20
ppm, ii) heat attenuated HD-1 protein control at a nominal
rate of 20 ppm, iii) and honey:water mixture only. Adult bees
were observed twice on the day of test initiation for mortality
and signs of toxicity and once each day thereafter. The study
was terminated at day 9 when the cumulative mortality in
the negative control group exceeded 20%.
characterized and assessed for potential effects against
earthworms. The earthworms were exposed to a single test
concentration of CryIA(b) protein in an artificial soil
substrate, and observed for mortality and signs of toxicity on
Day 7 and Day 14 of the test. The nominal test concentration
to which the earthworms were exposed was 200 mg CryIA(b)
protein/kg dry soil (mg/kg). A control group was maintained
concurrently. Four replicate test chambers were maintained
in each treatment and control group, with 10 worms in each
test chamber. The cumulative mortality percentage in the
treatment group was used to determine the LC50 value at
test termination. The no-observed-effect-concentration was
determined by visual examination of the mortality, body
weight and clinical observation data.
Conclusion
The difference in cumulative mortality between the
treatment and control groups was not statistically
significant. The LC50 for B.t.k. HD-1 protein in adult
honey bees (Apis mellifera) is greater than 20 ppm. The no
observed effect level was 20 ppm.
Reference
The 14 day LC50 value for earthworms exposed to
CryIA(b) protein in an artificial soil substrate was
determined to be greater than 200 mg CryIA(b) protein/kg
dry soil, the single concentration tested. The no-observedeffect-concentration was 200 mg CryIA(b) protein/kg dry
soil.
(Palmer and
Beavers, 1995)
21
MON 810
Monsanto Company
(Maggi and Sims,
1994a)
Table 1
Test Substance
Maize tissue
containing CryIA(b)1/
U.S.A.
Organism Tested
Collembola (Folsomia
candida)
Maize pollen
containing CryIA(b)/
U.S.A.
Cladoceran (Daphnia
magna)
1
Study Design
Collembola were exposed to maize plant tissue containing
CryIA(b) protein or to control maize plant tissue without the
CryIA(b) protein for 28 days. Three treatment levels of each
lyophilized tissue mixed with Brewer’s yeast, a standard
laboratory food for Collembola, were tested at a rate of 0.5,
5.0, 50% (wt:wt). The amended yeast diets were provided to
the Collembola ad libitum. At the end of the 28 day exposure
period, the number of Collembola surviving each treatment
was counted. Differences between treatments were
determined using standard parametric statistical procedures.
Daphnids were exposed to a single dose of maize pollen (100
mg test pollen/L) containing CryIA(b) protein. Two control
groups were included: a group exposed to a single dose of
maize pollen (100 mg control pollen/L) from conventional
maize and an assay control group of daphnids held in dilution
water only. Three replicate test chambers were maintained
for the test and control groups, with 10 neonate daphnids in
each chamber for a total of 30 neonate daphnids per test
concentration. The nominal concentration of pollen (both
control and test protein) was 100 mg/L based on U.S. EPA’s
guidance for limit tests.
Conclusion
The results of this study indicate that, even at very high
treatment levels, Collembola were not affected by chronic
exposure to CryIA(b) protein in plants. Following exposure
to the test materials in their diet for 28 days there was no
mortality and no reduction in the number of progeny.
Reference
The estimated 48-hour EC50 value for Daphnia magna
exposed to CryIA(b) protein in maize pollen was >100 mg
test pollen/L. There were no treatment-related effects
observed at the 100 mg test pollen/L limit concentration.
(Graves and
Swigert, 1997)
The summaries in this table use the terminology of the reports. CryIA(b) and B.t.k. HD-1 are synonymous with Cry1Ab.
22
MON 810
Monsanto Company
(Halliday, 1997)
For the Bt proteins tested in laboratory assays to date, including
Cry1Ab, potentially significant adverse effects have been
observed for only a very few non-target species that are closely
related to the target species (Mendelsohn et al., 2003; Romeis et
al., 2006). However, field studies conducted over the past decade
by industry and the academic community and reported in the
peer-reviewed literature on registered insect-protected crops that
produce a variety of Cry1A proteins, including Cry1Ab, have
demonstrated that these crops have no adverse effects on
biodiversity, tested populations of natural enemies, and other
ecologically important non-target arthropods (U.S. and other
world areas: (Daly and Buntin, 2005; Dively, 2005; Dively and
Rose, 2003; Head et al., 2001; Head et al., 2005; Lozzia et al.,
1998; Naranjo et al., 2005; Naranjo, 2005a; Naranjo, 2005b; Orr
and Landis, 1997; Pilcher et al., 1997; Pilcher et al., 2005; Torres
and Ruberson, 2005; Whitehouse et al., 2005) (E.U.: (Arpas et al.,
2005; Babendreier et al., 2004; Bakonyi et al., 2006; Bourguet et
al., 2002; Eckert et al., 2006; Freier et al., 2004; Heckmann et al.,
2006; Lang et al., 2004; Ludy and Lang, 2006a; Ludy and Lang,
2006b; Meissle et al., 2005; Romeis et al., 2004; Romeis et al.,
2006; Toth et al., 2004; Vercesi et al., 2006; Vojtech et al., 2005;
Volkmar and Freier, 2003; Wandeler et al., 2002).
Importantly, when expressed in Bt crops, even sensitive nontarget lepidopteran species have been shown to be exposed to
lower levels of Cry1A proteins compared to herbivores. These
reduced levels of exposure are too low to pose a significant risk to
populations of the sensitive non-target species (Hellmich et al.,
2001; Pleasants et al., 2001). For example, the impact of exposure
to pollen containing Cry1A proteins on lepidopteran species has
been evaluated in a number of empirical studies and several risk
assessments (Mendelsohn et al., 2003), and the risk has been
shown to be negligible.
MON 810 has been tested in vertebrate species including rats
and chickens, as well as other large animals. These studies
described in Section D.9.7, also demonstrate the lack of potential
adverse effects to nontarget vertebrate species.
In a laboratory study, which received considerable media
attention, Monarch butterfly larvae were reported to be
susceptible to the Cry1Ab protein found in Bt maize pollen (Losey
et al., 1999). Follow-up studies, however, demonstrated that
exposure of monarch and black swallowtail butterfly populations
to maize pollen is very low under field conditions (Dively et al.,
2004; Sears et al., 2001; Wraight et al., 2000). The researchers
concluded that: 1) maize pollen is heavy and does not travel far
from maize fields (>90% is deposited within 5 meters of the field
23
MON 810
Monsanto Company
perimeter) and 2) because maize pollen shed occurs only for a one
to two week period each growing season, exposure to maize
pollen is limited. Based on these facts, Dively et al. (2004) used a
risk assessment procedure and simulation model to estimate the
proportion of second-generation monarch butterflies affected by
Cry1Ab-expressing maize over the entire U.S. corn belt. Their
results showed that only 50% of the Monarch breeding
population was potentially exposed to Cry1Ab-containing maize
pollen. They determined that this level of exposure would result
in the risk of 0.6% additional mortality to monarch butterfly
larvae associated with long-term exposure to Bt maize pollen.
These results support the conclusion that MON 810 Bt pollen is
unlikely to pose any significant risk to the sustainability of
Monarch butterfly populations (Dively et al., 2004).
Recent opinions of EFSA’s Scientific Panel on Genetically
Modified Organisms (GMO Panel) related to the safeguard clause
invoked by Austria, Hungary and Greece on the authorized
genetically modified maize MON 810 (EFSA, 2004; EFSA, 2005;
EFSA, 2006), respectively) according to Article 23 of Directive
2001/18/EC, conclude that, in terms of risk to human health and
the environment, no new information affecting scientific evidence
was presented that would invalidate the risk assessment of
MON 810 established under Directive 90/220/EEC. The opinions
confirm EFSA’s conclusions with respect to the potential impact
of Cry1Ab protein, reflecting that MON 810 is unlikely to have
adverse effects on human and animal health or on the
environment.
The GMO panel has evaluated the relevance of the above
mentioned ‘safeguard clauses’ in the light of peer-reviewed
scientific data, which reflect the safety of the Cry1Ab protein on:
green lacewings (Bourguet et al., 2002; Dutton et al., 2002;
Dutton et al., 2003a; Dutton et al., 2003b; Romeis et al., 2004;
Romeis et al., 2006), non target non lepidopteran predators or
parasitoids (Bourguet et al., 2002; Candolfi et al., 2004; de la
Poza et al., 2005; Eizaguirre et al., 2006; Musser and Sehlton,
2003; Pons and Stary, 2003; Prutz and Dettner, 2004; Romeis et
al., 2004; Romeis et al., 2006; Siegfried et al., 2001), non target
lepidopterans (Dively et al., 2004; Eckert et al., 2006; Gatehouse
et al., 2002; Gathmann et al., 2006; Losey et al., 1999; Pons et al.,
2005; Rauschen et al., 2004; Sears et al., 2001; Yao et al., 2006;
Zangerl et al., 2001) and soil organisms such as Collembola,
earthworms and nematodes (Blackwood and Buyer, 2004; Evans,
2002; Heckmann et al., 2006; Motavalli et al., 2004; Saxena and
Stotzky, 2001; Vercesi et al., 2006).
24
MON 810
Monsanto Company
Mendelsohn et al. (2003) and the U.S. Environmental Protection
Agency (EPA) in its Biopesticides registration action document
(US EPA, 2001) also confirm that Bt maize does not pose
unreasonable adverse effects to non target wildlife or beneficial
invertebrates. This conclusion of safety is further supported by
assessments by leading European scientists (Lang et al., 2005;
Sanvido et al., 2006).
The results of extensive laboratory testing, long term field
evaluation, including extensive commercial use in a variety of
agricultural environments, suggests that no conclusive evidence
has yet been presented that currently released Bt crops are
causing significant direct or indirect adverse effects on NTO
populations. In addition, the available data do not indicate a
chain of effects that might result in long-term effects.
As expected, the Cry1Ab protein was shown to degrade rapidly in
soil, which confirms the absence of adverse effects on soil
microorganisms. The degradation rate of the Cry1Ab protein was
assessed by measuring the decrease in insecticidal activity of
MON 810 tissue material incubated in soil. The Cry1Ab protein,
as a component of the maize tissue, had an estimated DT50 (time
to 50% reduction of bioactivity) and DT90 (time to 90% reduction
of bioactivity) of 1.6 and 15 days, respectively (Sims and Holden,
1996). This measured rate of degradation in soil is comparable to
that reported for the Btk protein in genetically modified cotton
(Palm et al., 1994) and to the degradation rate reported for
microbial Bt products (Pruett et al., 1980; West, 1984; West et al.,
1984). This rapid degradation strongly supports the lack of
exposure of Cry1Ab on non-target organisms involved in the
decomposition function and on soil-dwelling organisms in
general. More recently, (Dubelman et al., 2005) showed that
Cry1Ab protein does not accumulate or persist in the
environment after 3 years of continuous use.
In conclusion, based on the well-characterised mode of action of
the Cry proteins, the selectivity of the Cry1Ab protein for certain
lepidopteran pests and the confirmation through studies showing
no adverse effects in diets it is concluded that the potential for
MON 810 to be hazardous to non-target organisms is negligible.
In conclusion, there is negligible risk for harmful effects of
MON 810
on
non-target
organisms
(vertebrates
and
invertebrates), either through direct or indirect interactions with
this maize or through contact with the newly expressed protein
Cry1Ab. Higher trophic interactions between non-target
organisms would also not be negatively affected. Therefore, any
25
MON 810
Monsanto Company
risks for significant indirect effects on the population levels of
non-target organisms in the receiving environment or their
functioning in below- and above-ground ecosystems in the
vicinity of the crop are equally negligible.
Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Expression of
Cry1Ab protein
Direct toxicity of
the expressed
protein on nontarget organisms
or indirect
population effects
Negligible
Negligible
(not applicable)
9.6
Effects on human health
2001/18/EC: “Possible immediate and/or delayed effects on human
health resulting from potential direct and indirect interactions of the
GMHP and persons working with, coming into contact with or in the
vicinity of the GMHP release(s).
effect
MON 810 was shown to be compositionally equivalent to
conventional maize, with no substantial differences from
conventional maize with respect to safety characteristics and
agronomic and phenotypic characteristics, except for the
introduced lepidopteran-protection trait, imparted by the
expression of Cry1Ab protein. Theoretically, potential toxicity
and/or allergenicity might be associated with newly expressed
proteins in a genetically modified crop. Therefore, the expression
of the Cry1Ab protein is a characteristic of the GMHP that may,
at least in theory, cause an adverse occupational health effect.
This is evaluated in this section.
If the introduced protein in MON 810 would have toxic or
allergenic potential, then this could cause a meaningful change
in the safety aspects of the handling of maize. Any change in the
26
MON 810
Monsanto Company
occupational health aspects of modified maize would need to be
understood compared to conventional maize.
Interaction of workers with maize
Since this renewal application is for cultivation of MON 810 in
the E.U. and use thereof as with any other maize, the persons
coming into contact with this maize will predominantly be
breeders, growers, persons involved in the handling, storage and
processing of maize grain, as well as people feeding the grain to
domestic animals. The likelihood to come into contact with
MON 810 and its grain is not different from the occupational
exposure to conventional maize, or to the parental maize
products.
Baseline occupational hazards associated with the handling of
maize
In general, allergic reactions to proteins can be elicited via
contact with these proteins in the environment or by ingesting
them through the diet. IgE-mediated reactions to environmental
allergens are typically less severe but occur at a higher
frequency than do IgE mediated food allergic reactions that are
estimated to affect 1-2 % of adults (Sicherer and Sampson, 2006).
Allergic reactions to maize proteins are exceedingly rare even
amongst atopic individuals who are thought to be more prone to
IgE
mediated
hypersensitivity
reactions
elicited
by
environmental or food allergens (Bock et al., 1978; Jones et al.,
1995; Pauls and Cross, 1998). Since occupational exposure to
maize is limited, the potential for developing allergic reactions to
maize grain is negligible.
Negligible potential for the Cry1Ab protein to cause occupational
health effects
MON 810 has no meaningful differences compared to
conventional maize except for the newly introduced trait
(protection against lepidopteran insect pests), which is imparted
by expression of the Cry1Ab protein. The safety of the introduced
trait to human health has been investigated extensively in
studies on the expressed Cry1Ab protein, as previously discussed
in Sections D.7.8 and D.7.9. The Cry1Ab protein has a history of
safe use. Its safety to humans has been established based on the
1) lack of acute toxicity as determined in a mouse gavage study,
2) rapid digestion in simulated gastric fluids, 3) lack of homology
with known protein toxins and 4) lack of homology with known
allergens. The Cry1Ab protein is also present at very low levels
in MON 810 (Section D.7.8.1).
27
MON 810
Monsanto Company
The extensive commercial experience with MON 810 did not
provide evidence that the occupational hazards associated with
the cultivation, storage, handling and processing of MON 810
are any different from conventional maize.
In conclusion, the likelihood for any adverse effects occurring in
humans as a result of their contact with this maize is no
different from conventional maize, as MON 810 contains the
Cry1Ab protein, which has negligible potential to cause any toxic
or allergenic effects in humans. Therefore, the risk of changes in
the occupational health aspects of this maize is negligible.
Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for the
marketing of
the GMHP
Expression of the
Cry1Ab protein
Adverse
occupational
health effects to
persons handling
MON 810,
resulting from
potential toxicity
or allergenicity of
the introduced
protein
Negligible
Negligible
(not applicable)
9.7
Effects on animal health
2001/18/EC: “Possible immediate and/or delayed effects on animal
health and consequences for the feed/food chain, resulting from
consumption of the GMO, if it is intended to be used as animal feed.”
effect
MON 810 is not substantially different from conventional maize
except for the introduced lepidopteran-protection trait. Based on
centuries of experience with conventional, domesticated maize in
Europe, there is negligible potential for maize to cause any
adverse health effects in livestock animals. Theoretically,
28
MON 810
Monsanto Company
potential toxicity or nutritional deficiency might result from
newly expressed proteins in the crop. Therefore, the expression
of the Cry1Ab protein is a characteristic of the GMHP that may,
at least in theory, cause an adverse effect. This is evaluated in
this section.
If the newly expressed Cry1Ab protein in MON 810 would have
toxic or allergenic potential or have an adverse effect on the
wholesomeness of this maize, then this could cause a meaningful
change in the nutritional or feed safety aspects of this maize.
This change may potentially result in an altered animal
performance such as a change in growth patterns, feed efficiency,
milk production or diminished health. However, any change in
the wholesomeness or health aspects of livestock fed the
modified maize would need to be understood compared to
conventional maize.
Exposure of domestic livestock
This renewal application is for cultivation of MON 810 in the
E.U. and use thereof as any other maize, including the use of
this maize as animal feed. Annually, more than 40 million
tonnes of maize grain are produced or imported into the E.U. of
which the majority (~80 %) is used for animal feed for livestock
animals destined for human consumption.
Grain from MON 810 is nutritionally equivalent to conventional
control as well as to maize varieties in commerce when fed to
rats (see Section D.7.8.4), broiler chickens (see Section D. 7.8.4;
(Gaines et al., 2001; Mireles et al., 2000; Rossi et al., 2005), swine
(Gaines et al., 2001; Piva et al., 2001; Weber et al., 2000), beef
cattle (Hendrix et al., 2000; Petty et al., 2001; Russell et al.,
2000a; Russell et al., 2001; Russell et al., 2000b) and lactating
dairy cattle (Donkin et al., 2003). Hence this maize is not
expected to be more or less attractive than conventional maize
for use as food or feed, for processing or as a food or feed
ingredient. Therefore, MON 810 is not expected to affect current
usage patterns of maize, but to replace a portion of the grain
from current maize hybrids such that their intake or use will
represent some fraction of the total products derived from maize.
Feed safety and wholesomeness
MON 810 is not substantially different from conventional maize
except for the intentionally introduced trait protection from
certain lepidopteran insect pests. The feed safety of the Cry1Ab
protein that confers this trait has been established previously by
29
MON 810
Monsanto Company
the data provided in MON 810 initial submission. On this basis,
MON 810 was approved for placing on the market in the E.U. in
1998 (Commission Decision, 1998).
As previously discussed, the Cry1Ab protein that is expressed in
MON 810 has a history of safe use (see section D.7.8.1).
Furthermore, mice did not show signs of toxicity in an acute oral
gavage study using doses orders of magnitude higher than
expected consumption levels from feed products containing or
consisting of MON 810. This lack of toxicity was expected based
on the absence of a toxic mechanism in mammals, the history of
exposure and the rapid degradation of each protein in simulated
human gastric fluids (see Section D.7.8.1).
The safety of maize containing this introduced protein was
further confirmed by a feeding study in broiler chickens and a
two-dose feeding study in the rat using MON 810 grain
containing diets (see Section D.7.8.4), demonstrating the absence
of any toxic or pleiotropic effects linked to the genetic
modification.
Feed wholesomeness was demonstrated in feeding studies with
poultry (see Section D. 7.8.4; (Gaines et al., 2001; Mireles et al.,
2000; Rossi et al., 2005), swine (Gaines et al., 2001; Piva et al.,
2001; Weber et al., 2000), beef cattle (Hendrix et al., 2000; Petty
et al., 2001; Russell et al., 2000a; Russell et al., 2001; Russell et
al., 2000b) and lactating dairy cattle (Donkin et al., 2003). No
differences were reported in performance, health, and meat and
milk quality measurements.
In conclusion, the likelihood of potential adverse effects in
animals fed on MON 810 and in humans, consuming those
animals, is negligible. Therefore, the risk of MON 810 for the
feed/food chain is also negligible.
30
MON 810
Monsanto Company
Characteristics
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Expression of
Cry1Ab protein
Adverse health
effects in the
feed/food chain or
wholesomeness
effects, resulting
from dietary
exposure of
livestock animals
to the introduced
Cry1Ab protein
Negligible
Negligible
(not applicable)
9.8
Effects on biogeochemical processes
2001/18/EC: “Possible immediate and/or delayed effects on
biogeochemical processes resulting from potential direct and indirect
interactions of the GMO and target and non-target organisms in the
vicinity of the GMO release(s).”
effect
Maize production in general is known to have indirect impacts on
biogeochemical processes through tillage, fertilizer application,
and establishment of a monoculture in a defined area. As
MON 810 was shown to be compositionally equivalent to nontransgenic maize and was not different in morphology,
development, yield, dissemination, stress susceptibility, plant
health and survival characteristics, there is no evidence that this
maize would be any different from conventional maize regarding
its direct influence on nutrient levels in the soil.
Expression of the newly introduced Cry1Ab protein is a
characteristic of the GMHP that could, at least in theory, cause
an adverse environmental effect on biogeochemical processes.
This is evaluated in this section.
31
MON 810
Monsanto Company
Potential toxicity of the Cry1Ab protein expressed by MON 810
to populations of decomposers, detritivores and soil microbial
processes in the receiving environment may affect the
biogeochemical processes that they are involved in, and change
nutrient recycling in the environment.
Throughout its lifecycle in the field, MON 810 interacts with a
spectrum of non-target organisms that are involved in the
biogeochemical processes of decomposition and nutrient recycling
in the soil. As biogeochemical processes are exceedingly complex,
such processes are best understood at a macroscopic, system level
than at specific, organismal level. The main functional groups of
non-target organisms that are relevant to the assessment of
potential adverse effects on biogeochemical processes include
decomposers of plant material and organic substances and
primary consumers feeding on organic debris (detritivores).
Primary consumers typically are macro-organisms that feed on
the detritus, i.e. the organic debris resulting from decomposition
of plant material. They further reduce the size of the detritus
particles through partial digestion, and, after defecation of the
particles, thereby enhance further decomposition. Detritivores
also provide aeration to the soil or litter material, thereby
improving the oxygen content in the soil and increasing
respiration of decomposers. Important examples of detritivores
are springtails (Collembola), millipedes and annelid worms.
Populations of soil-borne consumers are affected by plant
genotype on tillage practices, environmental conditions, previous
history of crops grown and the application of pesticides and
fertilisers. The Cry1Ab protein expressed in MON 810 was
demonstrated to have negligible risk for adverse environmental
effects through direct or indirect interactions with non-target
organisms, including representative detritivorous organisms that
are involved in the decomposition function in the soil.
Decomposers include bacteria and fungi (saprophytes) that break
down dead and decaying material, such as stubble plant
material, fresh litter remaining after harvest of the crop, smaller
detritus from more advanced decomposition, and humus. Dead
and decaying plant material contains important nutrients, e.g.
carbon which is released as carbon dioxide as a result of
microbial respiration, and nitrogen which is recycled by a range
of soil bacteria. Bacterial and fungal populations are critical to
maintaining soil health and quality. Soil microbial communities
that mediate biogeochemical processes are highly complex and
32
MON 810
Monsanto Company
are often characterized by high microbial diversity (Tiedje et al.,
1999). However, the diversity and abundance of these organisms
and hence their microbial processes are significantly affected by
biotic factors (community characteristics and dynamics), abiotic
factors (soil structure, clay type, moisture capacity,
environmental conditions, pH) and soil use (crop, tillage
practices, history of previously grown crops). Agricultural
practices such as fertilization and cultivation techniques may
also have profound effects on soil microbial populations, species
composition, colonization, and associated biochemical processes
(Alexander, 1961). Consequently, significant variation in
microbial populations is expected in the agricultural
environment.
Although the Cry1Ab protein present in decaying MON 810
material is considered to be a newly expressed protein in maize,
it is not a novel protein in the soil. The cry1Ab gene which was
used in this genetically modified maize was derived from the
genome of a common soil bacterium Bacillus thuringiensis subsp.
kurstaki.
The toxic mechanism of Cry1Ab protein has been thoroughly
characterised (see (Monsanto Company, 1995)) and was found to
be extremely specific to larvae of certain lepidopteran insect
pests. Consequently, the potential for activity of this protein
towards microorganisms is negligible. In addition, as expected,
the Cry1Ab protein was shown to degrade rapidly in soil, which
confirms the absence of adverse effects on soil microorganisms.
The degradation rate of the Cry1Ab protein was assessed by
measuring the decrease in insecticidal activity of MON 810 tissue
incubated in soil (Section D.9.5). This rapid degradation strongly
supports the lack of exposure of Cry1Ab on non-target organisms
involved in the decomposition function and on soil-dwelling
organisms in general.
Finally,
extensive
commercial
experience
with
the
commercialisation of various Cry1Ab-expressing insect-protected
crops has not revealed any adverse or undesirable effects on
biogeochemical processes or soil fertility.
It is highly unlikely that there is any difference between
MON 810 and conventional maize with respect to its direct
influence on soil nutrient levels and key processes. Furthermore,
it is highly unlikely that the direct or indirect interaction
between this maize and decomposers or detritivores in the
receiving environment would cause any immediate or delayed
33
MON 810
Monsanto Company
adverse effects on the decomposition and nutrient recycling
functions in the soil.
In conclusion, the environmental risk of adverse effects on
biogeochemical processes, caused by the interaction of MON 810
with target and non-target organisms in the soil, is negligible.
Characteristic
s of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the
marketing of
the GMHP
Expression of
Cry1Ab protein
Adverse effects on
nutrient cycles in
the soil, resulting
from potential
adverse effects of
the introduced
protein on target
or non-target
organisms
involved in
biogeochemical
processes
Negligible
Negligible
(not applicable)
9.9
Impact of the specific
harvesting techniques
cultivation,
management
and
2001/18/EC: “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.”
effect
No characteristics could be identified which may cause an
adverse environmental effect. Compared to conventional maize,
any new characteristics in MON 810 are limited to the
lepidopteran-protection trait.
If the introduced trait(s) in a GMHP would have altered the
plant’s agronomic or environmental characteristics in such a way
that the crop would require the application of specific cultivation,
management or harvesting techniques – different from those
34
MON 810
Monsanto Company
available in current agricultural practices – then those novel or
specific techniques need to be evaluated for their potential direct
or indirect adverse environmental effects.
If different cultivation, management or harvesting techniques
would become necessary in order to successfully grow this maize,
then such new techniques could affect, at least in theory, the
biotic or abiotic characteristics of the receiving environment
wherein the crop is grown. The need for new management
techniques and their potential adverse environmental effects are
evaluated in this section.
However, for any effect from changing management practices to
be meaningful for a given crop, it would have to be understood in
comparison with the inherent variability of biotic and abiotic
factors in the agronomic environment, e.g. as they are affected by
the type of crop that is planted. Moreover, the adversity of such a
change needs to be considered in the light of the tremendous
environmental impact of today’s agricultural practices in general
(i.e. cultivation of large monocultures in a well-prepared, manmade environment; where crop plants concertedly develop from
seed to flowering stage in a relatively short time span; protected
throughout the season from potentially harmful interactions with
other organisms by multiple applications of various chemicals
(some of which have broad-spectrum or non-target activity); with
the adult plants to be cut, finally, and removed from the field at
harvest).
As MON 810 is equivalent to conventional maize, except for the
introduced lepidopteran-protection trait, all the agronomic
practices currently used to grow maize in the E.U. remain
applicable for growing MON 810 and no new or specific
techniques for cultivation, management and harvesting are
necessary.
•
No specific cultivation techniques are required to
grow MON 810. For instance, traditional crop rotational
practices, planting regimes for maize, techniques for soil
preparation (tillage), maize drilling techniques and all
technical equipment remain applicable.
• Similarly, no new or specific crop management techniques are
required for MON 810. All the conventional management
techniques to cultivate maize remain at the farmer’s
disposition, e.g. application of fertiliser, irrigation techniques,
35
MON 810
Monsanto Company
mechanical operations or the use of approved plant protection
products for disease, insect pest and weed control.
• Finally, no changes in harvesting techniques are required.
Traditional harvesting equipment as well as post-harvest
storage techniques and conditions remain applicable.
To further clarify the absence of any requirements for specific
crop management techniques, we note that the importance and
aim of the basic management technique of removal of harmful
insect pests from the field in order to achieve optimal yield of the
crop, is not different between MON 810 and any other maize. The
lepidopteran-protection trait allows the farmer to effectively
control some of his most important insect pests, while avoiding
potential operator exposure to the insecticides traditionally used
to control these pests and without having to perform the costly
field operations. Therefore, cultivation of MON 810 instead of
conventional maize does not change any basic management
technique in maize as such, but gives growers more flexibility to
apply the existing tools for management, while creating at the
same time new opportunities to grow maize in a more
sustainable way (e.g. reduced tillage or integrated pest
management).
In conclusion, in comparison to any other maize, no typical
characteristics of the genetically modified plant could be
identified, which may cause adverse effects on the environment
through a need to change management practices. Therefore, the
environmental impact of farming practices to grow MON 810 in
the E.U. is considered no different from any other maize.
It is actually expected that the production of MON 810 will
positively impact current agronomic practices in maize and
provide benefits to farmers and the environment in the E.U. The
benefits of planting insect-protected maize include: 1) a reliable
means to control the target lepidopteran maize pests; 2) control
of target insects while maintaining beneficial species; 3) reduced
use of chemical insecticides (Rice and Pilcher, 1999); 4) reduced
applicator exposure to chemical pesticides; 5) good fit with
integrated pest management (IPM) and sustainable agricultural
systems; 6) reduced fumonisin mycotoxin levels in maize kernels
(Masoero et al., 1999; Munkvold et al., 1999); and 7) no additional
labour or machinery requirements, allowing both large and small
growers to maximize hybrid yields. MON 810 can offer the abovementioned agronomic, environmental, and therefore also societal
benefits from both these traits in maize.
36
MON 810
Monsanto Company
In order to secure the valuable agronomic and other benefits of
insect-protected maize on a longer term, a harmonised Insect
Resistance Management (IRM) stewardship programme was
developed, aiming to delay the onset and development of possible
resistance in target insect species (see harmonised Insect
Resistance Management plan in Appendix 1 of the current
dossier). This stewardship plan is part of a larger stewardship
effort in the E.U., which is currently being implemented for
various insect-protected Bt maize varieties.
considered applicable.
Characteristic
of the GMHP
which may
cause an
adverse effect
Potential
consequence of
the adverse
effect, if it
occurs
Likelihood of
occurrence of
the potential
adverse effect
Estimation of the
risk posed by the
characteristic of
the GMHP
Risk
Management
strategy for
the marketing
of the GMHP
Introduced
protection from
targeted
lepidopteran insect
pests
Potential for adverse
effects to biotic or
abiotic factors in the
environment,
resulting from the
application of novel
or specific farming
techniques,
potentially required
to cultivate, manage
or harvest MON 810
Negligible
Negligible
(not applicable)
37
MON 810
Monsanto Company
References
AGPM. (1999) Dispersion du pollen en production de mais consommation. Etude
réalisée dans le cadre de comité de biovigilance.
Alexander, M. (1961) Introduction to soil microbiology. John Wiley and Sons.
Aronson, A.I. and Shai, Y. (2001) Why Bacillus thuringiensis insecticidal toxins
are so effective: unique features of their mode of action. FEMS Microbiol.
Lett., 195, 1-8.
Arpas, K., Toth, F. and Kiss, J. (2005) Foliage-dwelling Arthropods in Bttransgenic and Isogenic Maize: A comparison through spider web analysis.
Acta Phytopathologica et Entmologica Hungarica, 40, 347-353.
Babendreier, D., Kalberer, N., Romeis, J., Fluri, P. and Bigler, F. (2004) Pollen
consumption in honey bee larvae: a step forward in the risk assessment of
transgenic plants. Apidologie, 35, 293-300.
Bakonyi, G., Szira, F., Kiss, I., Villanyi, I., Seres, A. and Szekacs, A. (2006)
Preference tests with collembolas on isogenic and Bt maize. Eur. J. Soil
Biol., 42, S132 - S135.
Blackwood, C.B. and Buyer, J.S. (2004) Soil microbial communities associated
with Bt and Non-Bt Corn in three soils. J. Environ. Qual., 33, 832-836.
Bock, S.A., Lee, W.Y., Remigio, L.K. and May, C.D. (1978) Studies of
hypersensitivity reactions to foods in infants and children. J. Allergy Clin.
Immunol., 62, 327-334.
Bodet, J.M., Straebler, M. and Broucqsault, L.M. (1994) Type de jachère et
couvert. Receuil des communications du colloque "Jachères 94", 19-41.
Bourguet, D., Chaufaux, J., Micoud, A., Delos, M., Naibo, B., Bombarde, F.,
Marque, G., Eychenne, N. and Pagliari, C. (2002) Ostrinia nubilalis
parasitism and the field abundance of non-target insects in transgenic
Bacillus thuringiensis corn (Zea mays). Environ. Biosafety Res., 1, 49-60.
Broderick, N., Raffa, F.K. and Handelsman, J. (2006) Midgut bacteria required
for Bacillus thuringiensis insecticidal activity. proc. Natl. Acad. Sci., 103,
15196-15199.
Candolfi, M.P., Brown, K., Grimm, C., Reber, R. and Schmidli, H. (2004) A
faunistic approach to assess potential side effects of genetically modified Bt
corn on non-target arthropods under field conditions. Biocontrol Science
and Technology, 14, 129-170.
Cantwell, G.E., Lehnert, T. and Fowler, J. (1972) Are biological insecticides
harmful to the honey bee? American bee journal, 294-296.
Commission Decision. (1998) Commission Decision of 22 April 1998 concerning
the placing on the market of genetically modified maize (Zea mays L. line
MON 810), pursuant to Council Directive 90/220/EEC. Official Journal.
38
MON 810
Monsanto Company
Crickmore, N., Zeigler, D.R., Feitelson, J., Schnepf, E., Van Rie, J., Lereclus, D.,
Baum, J. and Daen, D.H. (1998) Revision of the nomenclature for the
Bacillus thuringiensis pesticidal crystal proteins. Microbiology and
Molecular Biology Reviews, 62, 807-813.
Crickmore, N., Zeigler, D.R., Schnepf, E., Van Rie, J., Lereclus, D., Baum, J.,
Bravo, A. and Dean, D.H. (2005) Bacillus thuringiensis toxin
nomenclature.
http://www.lifesci.sussex.ac.uk/Home/Neil_Crickmore/Bt/.
Daly, T. and Buntin, G.D. (2005) Effect of Bacillus thuringiensis transgenic corn
for Lepidopteran control on nontarget arthropods. Environ. Entomol., 34,
1292-1301.
de la Poza, M., Pons, X., Farinos, G.P., Lopez, C., Ortego, F., Eizaguirre, M.,
Castanera, P. and Albajes, R. (2005) Impact of farm-scale Bt maize on
abundance of predatory arthropods in Spain. Crop Protection, 24, 677-684.
Devos, Y., Reheul, D. and De Schrijver, A. (2005) Review: The co-existence
between transgenic and non-transgenic maize in the European Union: a
focus on pollen flow and cross fertilization. Environ. Biosatety Res., 4, 7187.
Dively, G.P. (2005) Impact of transgenic VIP3A x Cry1Ab Lepidopteran-resistant
field corn on the nontarget arthropod community. Environ. Entomol., 34,
1267-1291.
Dively, G.P. and Rose, R. (2003) Effects of Bt transgenic and conventional
insecticide control on the non-target natural enemy community in sweet
corn. Proceedings of the 1st International Symposium on Biological Control
of Arthropods., 265-274.
Dively, G.P., Rose, R., Sears, M.K., Richard, L., Hellmich, R.L., Stanley-Horn,
D.E., Calvin, D.D., Russo, J.M. and Anderson, P.L. (2004) Effects on
monarch butterfly lavae (Lepidoptera: Danaidae) after continuous
exposure to Cry1Ab-expressing corn during anthesis. Environmental
Entomology, 33, 1116-1125.
Donkin, S.S., Velez, J.C., Totten, A.K., Stanisiewski, E.P. and Hartnell, G.F.
(2003) Effects of feeding silage and grain from glyphosate-tolerant or
insect-protected corn hybrids on feed intake, ruminal digestion, and milk
production in dairy cattle. J. Dairy Sci., 86, 1780-1788.
Dubelman, S., Ayden, B.R., Bader, B.M., Brown, C.R., Jiang, C. and Vlachos, D.
(2005) Cry1Ab Protein does not persist in soil after 3 years of sustained Bt
Corn use. Environm. Entomol., 34, 915-921.
Dulmage, H.T. (1981) Insecticidal activity of isolates of Bacillus thuringiensis and
their potential for pest control. Microbial control of pests and plant diseases
1970-1980. Burger, H.D., Vol. 11, pp. 193-222.
Dutton, A., Klein, H., Romeis, J. and Bigler, F. (2002) Uptake of Bt-toxin by
herbivores on transgenic maize and consequences for the predator
Chrysoperla carnea. Ecol. Entomol., 27, 441-447.
39
MON 810
Monsanto Company
Dutton, A., Klein, H., Romeis, J. and Bigler, F. (2003a) Prey-mediated effects of
Bacillus thuringiensis spray on the predator Crysoperla carnea in maize.
Biological Control, 26, 209-215.
Dutton, A., Romeis, J. and Bigler, F. (2003b) Assessing the risks of insect
resistant transgenic plants on entomophagous arthropods: Bt-maize
expressing Cry1Ab as a case study. BioControl, 48, 611-636.
Eckert, J., Schuphan, I., Hothorn, L.A. and Gathmann, A. (2006) Arthropods on
maize ears for detecting impacts of Bt maize on non target organisms.
Environmental Entomology, 35, 554-560.
EFSA. (2004) Opinion of the Scientific Panel on genetically modified organisms
on a request from the Commission related to the Austrian invoke of Article
23 of Directive 2001/18/EC (Question N° EFSA-Q-2004-062). The EFSA
Journal, 78, 1-13.
EFSA. (2005) Opinion of the scientific panel on genetically modified organisms on
a request from the Commission related to the safeguard clause invoked by
Hungary according to Article 23 of Directive 2001/18/EC. The EFSA
Journal, 228, 1-14.
EFSA. (2006) Opinion of the Scientific Panel on genetically modified organisms
on a request from the Commission related to the safeguard clause invoked
by Greece according to Article 23 of Directive 2001/18/EC and to Article 18
of Directive 2002/53/EC. The EFSA Journal, 411, 1-26.
Eizaguirre, M., Albajes, R., Lopez, C., Eras, J., Lumbierres, B. and Pons, X.
(2006) Six years after the commercial introduction of Bt maize in Spain:
field evaluation, impact and future prospects. Transgenic Research, 15.
Emmerling, M., Chandler, D. and Sandeman, M. (2001) Molecular cloning of
three cDMAs encoding aminopeptidases from the midgut of Helicoverpa
punctigera, the Australian native budworm. Insect Biochem. Mol. Biol., 31,
899-907.
Evans, H.F. (2002) Environmental impact of Bt exudates from roots of genetically
modified plants. Defra-report, EPG 1/5/156.
Flexner, J.L., Lighthart, B. and Croft, B.A. (1986) The effects of microbial
pesticides on non-target, beneficial arthropods. Agriculture, ecosystems and
environment, 16, 203-254.
Freier, B., Schorling, M., Traugott, M., Juen, A. and Volkmar, C. (2004) Results
of a 4-year plant survey and pitfall trapping in Bt maize and conventional
maize fields regarding the occurrence of selected arthropod taxa.
IOBC/wprs Bulletin, 27.
Gaines, A.M., Allee, G.L. and Ratliff, B.W. (2001) Nutritional evaluation of Bt
(MON810) and Roundup Ready corn compared with commercial hybrids in
broilers. Poultry Science, 80, 51.
40
MON 810
Monsanto Company
Garner, K., Hiremath, S., Lethoma, K. and Valaitis, A.P. (1999) Cloning and
complete sequence characterization of two gypsy moth aminopeptidase-N
cDNAs, including the receptor for Bacillus thuringiensis Cry1Ac toxin.
Insect Biochem. Mol. Biol., 29, 527-535.
Gatehouse, A.M.R., Ferry, N. and Raemaekers, R.J.M. (2002) The case of the
monarch butterfly: a verdict is returned. TRENDS in Genetics, 18, 249251.
Gathmann, A., Wirooks, L., Hothorn, L.A., D., B. and Schuphan, I. (2006) Impact
of Bt maize pollen (MON 810) on lepidopteran larvae living on
accompanying weeds. Molecular Ecology, 15, 2677-2685.
Gill, M. and Ellar, D. (2002) Transgenic Drosophila reveals a functional in vivo
receptor for the Bacillus thuringiensis toxin Cry1Ac1. Insect Molecular
Biology, 11, 619-625.
Gonzalez-Nunez, M., Ortego, F. and Castanera, P. (2000) Susceptibility of
Spanish populations of the corn borers Sesamia nonagrioides (Lepidoptera:
Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) to a Bacillus
thuringiensis endotoxin. J. Economic Entomology, 93, 459-463.
Graves, W.C. and Swigert, J.P. (1997) Corn pollen containing the Cry1A(b)
protein: a 48-hour static-renewal acute toxicity test with the cladoceran
(Daphnia magna). Monsanto Technical Report, WL-96-322.
Hallauer, A.R. (1995) Potential for outcrossing and weediness of genetically
modified insect protected corn. .
Halliday, W.R. (1997) Chronic exposure of Folsomia candida to corn tissue
expressing Cry1A(b) protein. Monsanto Technical Report, XX-97-064.
Halsey, M.E., Remund, K.M., Davis, C.A., Qualls, M., Eppard, P.J. and Berberich,
S.A. (2005) Isolation of maize from pollen-mediated gene glow by time and
distance. Crop Sci., 45, 2172-2185.
Hansen, L. (1999) Non-target effects of Bt corn pollen on the monarch butterfly
(Lepidoptera Danaidae). Abstracts from the 54th Annual meeting North
Central Branch of the Entomological Society of America.
Head, G., Brown, C.R., Groth, M.E. and Duan, J.J. (2001) Cry1Ab protein levels
in phytophagous insects feeding on transgenic corn: implications for
secondary exposure risk assessment. Entomologia Experimentalis et
Applicata, 99, 37-45.
Head, G., Moar, W., Eubanks, M., Freeman, B., Ruberson, J., Hagerty, A. and
Turnipseed, S. (2005) A multiyear, large-scale comparison of arthropod
populations on commercially managed Bt and non-Bt cotton fields.
Heckmann, L.H., Griffiths, B., Caul, S., Thomson, J., Pusztai-Carey, M., Moar,
W.J., Andersen, M.N. and Krogh, P.H. (2006) Consequences for
Protaphorura armata (Collembola: Onychiuridae) following exposure to
genetically modified Bacillus thuringiensis (Bt) maize and non-Bt maize.
Environmental Pollution, 142, 212-216.
41
MON 810
Monsanto Company
Hellmich, R.L., Siegfried, B.D., Sears, M.K., Stanley-Horn, D.E., Daniels, M.J.,
Mattila, H.R., Spencer, T., Bidne, K.G. and Lewis, L.C. (2001) Monarch
larvae sensitivity to Bacillus thuringiensis-purified proteins and pollen.
Proc. Natl. Acad. Sci., 98, 11925-11930.
Hendrix, K.S., Petty, A.T. and Lofgren, D.L. (2000) Feeding value of whole plant
silage and crop residues from Bt or normal corns. J. Anim. Sci, 78, 273.
Hicks, D.A. and Thomison, P.R. (2004) Corn Management. Corn: Origin, History,
Technology, and Production, Chapter 3.2, 481-522.
Hofmann, C., Luthy, P., Hutter, R. and Pliska, V. (1988a) Binding of the delta
endotoxin from Bacillus thuringiensis to brush- border membrane vesicles
of the cabbage butterfly (Pieris brassicae). Eur J Biochem, 173, 85-91.
Hofmann, C., Vanderbruggen, H., Hoefte, H., Van Rie, J., Jansens, S. and Van
Mellaert, H. (1988b) Specificity of Bacillus thuringiensis delta-endotoxins
is correlated with the presence of high-affinity binding sites in the brush
border membrane of target insect midguts. Proc. Natl. Acad. Sci. USA, 85,
7844-7848.
Hoxter, K.A. and Lynn, S.P. (1992a) Activated Btk HD-1 protein: a dietary
toxicity study with green lacewing larvae. Monsanto Technical Report, WL92-155.
Hoxter, K.A. and Lynn, S.P. (1992b) Activated Btk HD-1 protein: a dietary
toxicity study with ladybird beetles. Monsanto Technical Report, WL-92156.
Hoxter, K.A. and Lynn, S.P. (1992c) Activated Btk HD-1 protein: a dietary
toxicity study with parasitic hymenoptera (Brachymeria intermedia).
Monsanto Technical Report, WL-92-157.
Jones, S.M., Magnolfi, C., Cooke, S.K. and Sampson, H.A. (1995) Immunologic
cross-reactivity among cereal grains and grasses in children with atopic
food hypersensitivity. J. Allergy Clin. Immunol., 96, 341-351.
Klausner, A. (1984) Microbial insect control. Biotechnology, 408-419.
Krieg, A. and Langenbruch, G.A. (1981) Susceptibility of arthropod species to
Bacillus thuringiensis. In Burges, H.D. (ed.) Microbial control of pests and
plant diseases 1970-1980, pp. 837-896.
Lang, A., Arndt, M., Beck, R., Bauchenss, J., Pommer, G. and Arndt, M. (2005)
Monitoring der Umwelwirkungen des Bt-Gens. Forschungsprojekt im
Auftrag des Bayerischen Staatsministeriums für Uwelt, Gesundheit und
Verbraucherschutz.
http://www.LfL.bayern.de/publikationen/daten/schriftenreihe_url_l_28.p
df.
Lang, A., Ludy, C. and Vojtech, E. (2004) Dispersion and deposition of Bt maize
pollen in field margins. Zeitschrift für Pflanzenkrankheiten und
Pflanzenschutz (J. Plant Diseases and Protection), 111, 417-428.
Losey, J.E., Rayor, L.S. and Carter, M.E. (1999) Transgenic pollen harms
monarch larvae. Nature, 399, 214.
42
MON 810
Monsanto Company
Lozzia, G., Furlanis, C., Manachini, B. and Rigamonti, L. (1998) Effects of Bt corn
on Rhopalosiphum padi L. (Rhynchota Aphididae) and on its predator
Chrysoperla carnea Stephen (Neuroptera Chrysopidae). Boll. Zool. Agraria
Bachicol., 30, 153-164.
Ludy, C. and Lang, A. (2006a) A 3-year field-scale monitoring of foliage dwelling
spiders (Araneae) in transgenic Bt maize fields and adjacent field margins.
Biological Control, 38.
Ludy, C. and Lang, A. (2006b) Bt maize pollen exposure and impact on the
garden spider, Araneus diadematus. Entomologia Experimentalis et
Applicata, 118, 145-156.
Luna, V., Figueroa, J.M., Baltazar, B.M., Gomez, R.L., Townsend, R. and
Schoper, J.B. (2001) Maize pollen longevity and distance isolation
requirement for effective pollen control. Crop Sci., 41, 1551-1557.
Ma, B.L., Subedi, K.D. and Reid, L.M. (2004) Crop ecology, management &
quality - extent of cross-fertilization in maize by pollen from neighboring
transgenic hybrids. Crop Sci, 44, 1273-1282.
MacIntosh, S.C., Stone, T.B., Sims, S.R., Hunst, P.L., Greenplate, J.T., Marrone,
P.G., Perlak, F.J., Fischhoff, D.A. and Fuchs, R.L. (1990) Specificity and
efficacy of purified Bacillus thuringiensis proteins against agronomically
important insects. J. Invertebr. Pathol., 56, 258-266.
Maggi, V.L. and Sims, S.R. (1994a) Evaluation of the dietary effects of purified
B.t.k. endotoxin proteins on honey bee adults. Monsanto Technical Report,
IRC-91-ANA-12.
Maggi, V.L. and Sims, S.R. (1994b) Evaluation of the dietary effects of purified
B.t.k. endotoxin proteins on honey bee larvae. Monsanto Technical Report,
IRC-91-ANA-13.
Mamarot, J. and Rodriguez, A. (1994) Etude du salissement des sols par la
jachère en région Midi-Pyrénées. Recueil des communications du colloque
"Jachères", 107-111.
Masoero, F., Moschini, M., Rossi, F., Prandini, A. and Pietri, A. (1999) Nutritive
value, mycotoxin contamination and in vitro rumen fermentation of normal
and genetically modified corn (Cry1A(B)) grown in northern Italy.
Maydica, 44, 205-209.
Meissle, M., Vojtech, E. and Poppy, G.M. (2005) Effects of Bt maize-fed prey on
the generalist predator Poecilus cupreus L. (Coleoptera: Carabidae).
Melin, B.E. and Cozzi, E.M. (1990) Safety to nontarget invertebrates of
lepidopteran strains of Bacillus thuringiensis and their Beta exotoxins.
Safety of microbial insecticides, 149-167.
Mendelsohn, M., Kough, J., Vaituzis, Z. and Matthews, K. (2003) Are Bt crops
safe? Nature Biotechnology, 21, 1003-1009.
Messeguer, J. (2003) Gene flow assessment in transgenic plants. Review of Plant
Biotechnology and Applied Genetics, 73, 201-212.
43
MON 810
Monsanto Company
Mireles, J., A., Kim, S., Thompson, R. and Amundsen, B. (2000) GMO (Bt) corn is
similar in composition and nutrient availability to broilers as non-GMO
corn. Poult. Sci., 79, 65-66.
Monsanto Company. (1995) Submission to the French Commission du Génie
Biomoléculaire. Application to place on the market genetically modified
higher plants: insect-protected maize (MON810). Monsanto report.
Motavalli, P.P., Kremer, R.J., Fang, M. and Means, N.E. (2004) Impact of
genetically modified crops and their management on soil microbially
mediated plant nutrient transformations. Journal of Environmental
Quality, 33, 816-824.
Munkvold, G.P., Hellmich, R.L. and Rice, L.G. (1999) Comparison of fumonisin
concentrations in kernels of transgenic Bt maize hybrids and nontransgenic hybrids. Plant disease, 83, 130-138.
Musser, F.R. and Sehlton, A.M. (2003) Bt sweet corn and selective insecticides:
impacts on pests and predators. J. Econ. Entomol., 96, 71-80.
Naranjo, S., Head, G. and Dively, G. (2005) Field studies assessing arthropod
non-target effects in Bt transgenic crops. Environ. Entomol., 34, 11781180.
Naranjo, S.E. (2005a) Long-term assessment of the effects of transgenic Bt cotton
on the abundance of nontarget arthropod natural enemies. Environ.
Entomol., 34, 1193-1210.
Naranjo, S.E. (2005b) Long-term assessment of the effects of transgenic Bt cotton
on the function of the natural enemy community. Environm. Entomol., 34,
1211-1223.
Noteborn, H.P. and Kuiper, H.A. (1994) Safety assessment strategies for
genetically modified plant products: a case study of Bacillus thuringiensistoxin tomato. Biosafety of foods derived by modern biotechnology, BATS.
OECD. (2003) Consensus Document on the Biology of Zea Mays Subsp. Mays
(Maize). http://www.oecd.org/.
Orr, D.R. and Landis, D.A. (1997) Oviposition of European corn borer
(Lepidoptera: Pyralidae) and impact of natural enemy populations in
transgenic versus isogenic corn. J. Econ. Entomol., 90, 905-909.
Palm, C.J., Donegan, K., Harris, D. and Seidler, R.J. (1994) Quantification in soil
of Bacillus thuringiensis var. kurstaki delta-endotoxin from transgenic
plants. Molecular Ecology, 3, 145-151.
Palmer, S.J. and Beavers, J.B. (1995) Cry1A(b) insecticidal protein: an acute
toxicity study with the earthworm in an artificial soil substrate. Monsanto
Technical Report, WL-95-281.
Pauls, J.D. and Cross, D. (1998) Food-dependent exercise induced anaphylaxis to
corn. J. Allergy Clin. Immunol., 101, 853-854.
44
MON 810
Monsanto Company
Petty, A.T., Hendrix, K.S., Stanisiewski, E.P. and Hartnell, G.F. (2001) Feeding
value of Bt corn grain compared with its parental hybrid when fed in beef
cattle finishing diets. J. Anim. Sci., 102, 79.
Pilcher, C.D., Obrycki, J.J., Rice, M.E. and Lewis, L.C. (1997) Preimaginal
development, survival and field abundance of insect predators on
transgenic Bacillus thuringiensis Corn. Biological Control, 26, 446-454.
Pilcher, C.D., Rice, M.E. and Obrycki, J.J. (2005) Impact of transgenic Bacillus
thuringiensis corn and crop phenology on five nontarget arthropods.
Piva, G., Morlacchini, M., Pietri, A., Piva, A. and Casadei, G. (2001) Performance
of weaned piglets fed insect-protected (MON810) or near isogenic corn. J.
Animal Sci., 79, 106, Abstract 441.
Pleasants, J.M., Hellmich, R.L., Dively, G.P., Sears, M.K., Stanley-Horn, D.E.,
Mattila, H.R., Foster, J.E., Clark, T.L. and Jones, G.D. (2001) Corn pollen
deposition on milkweeds in and near cornfields. Proc. Natl. Acad. Sci.
USA, 98, 11919-11924.
Pons, X., Lumbierres, B., Lopez, C. and Albajes, R. (2005) Abundance of nontarget pests in transgenic Bt-maize: A farm scale study. Eur. J. Entomol.,
102, 73-79.
Pons, X. and Stary, P. (2003) Spring aphid-parasitoid (Hom. Aphididae, Hym.
Braconidae) association and interactions in a Mediterranean arable crop
ecosystem, including Bt maize. J. Pest Sci., 76, 133-138.
Pruett, C.J.H., Burges, H.D. and Wyborn, C.H. (1980) Effect of exposure to soil on
potency and spore viability of Bacillus thuringiensis. J. Invertebr. Pathol.,
35, 168-174.
Prutz, G. and Dettner, K. (2004) Effect of Bt corn leaf suspension on food
consumption by Chilo partellus and life history parameters of its
parasitoid Cotesia flavipes under laboratory conditions. Entomologia
Rauschen, S., Eckert, J., Grathmann, A. and Schuphan, I. (2004) Impact of
growing Bt-maize on cicadas: diversity, abundance and methods.
IOBC/WPRS Bulletins "Ecological risk of GMO's, 27, 137-142.
Rice, M.E. and Pilcher, C.D. (1999) Bt corn and insect resistance management:
farmer perceptions and educational opportunities. A poster presented at the
1999 meeting of the Entomological Society of America.
Romeis, J., Dutton, A. and Bigler, F. (2004) Bacillus thuringiensis toxin (Cry1Ab)
has no direct effect on larvae of the green lacewing Chrysoperla carnea.
Journal of Insect Physiology, 50, 175-183.
Romeis, J., Meissle, M. and Bigler, F. (2006) Transgenic crops expressing Bacillus
thuringiensis toxins and biological control. Nature Biotechnology, 24, 63-71.
Rossi, F., Morlacchini, M., Fusconi, G., Pietri, A., Mazza, R. and Piva, G. (2005)
Effect of Bt corn on broiler growth performance and fate of feed-derived
DNA in the digestive tract. Poultry Science, 84, 1022-1030.
45
MON 810
Monsanto Company
Russell, J.R., Farnham, D., Berryman, R.K., Hersom, M.J., Pugh, A. and Barrett,
K. (2000a) Nutritive value of the crop residues from bt-corn hybrids and
their effects on performance of grazing beef cows. . Iowa State University,
Iowa State, pp. 56-61.
Russell, J.R., Hersom, M.J., Haan, M.M., Kruse, M.L. and Morrical, D.G. (2001)
Effects of grazing crop residues from bt-corn hybrids on pregnant beef
cows. J. Anim. Sci., 79, 74-75.
Russell, J.R., Hersom, M.J., Pugh, A., Barrett, K. and Farnham, D. (2000b)
Effects of grazing crop residues from bt-corn hybrids on the performance of
gestating beef cows. J. Anim. Sci., 78, 79-80.
Sacchi, V.F., Parenti, P., Hanozet, G.M., Giordana, B., Lüthy, P. and
Wolfersberger, M.G. (1986) Bacillus thuringiensis toxin inhibits K+ gradient-dependent amino acid transport across the brush border
membrane of Pieris brassicae midgut cells. FEBS Letters, 204, 213-218.
Sanvido, O., Stark, M., Romeis, J. and Bigler, F. (2006) Ecological impacts of
genetically modified crops. Experiences from ten years of experimental
field research and commercial cultivation. Agroscope Reckenholz-Tänikon
Research Station ART, Reckenholzstrasse 191, CH-8046 Zürich.
Saxena, D. and Stotzky, G. (2001) Bacillus thuringiensis (Bt) toxin released from
root exudates and biomass of Bt corn has no apparent effect on
earthworms, nematodes, protozoa, bacteria, and fungi in soil. Soil Biol.
and Biochem., 33, 1225-1230.
Sears, M.K., Hellmich, R.L., Stanley-Horn, D.E., Oberhauser, K.S., Pleasants,
J.M., Mattila, H.R., Siegfried, B.D. and Dively, G.P. (2001) Impact of Bt
corn pollen on Monarch butterfly populations: a risk assessment. Proc.
Natl. Acad. Sci., 98, 11937-11942.
Shaw, R.H. (1988) Climate requirement. Corn and Corn Improvement, 609-638.
Siegfried, B.D., Zoerb, A.C. and Spencer, T. (2001) Development of European corn
borer larvae on event 176 Bt corn: influence on survival and fitness.
Entomologia Experimentalis et Applicata, 100, 15-20.
Sims, S.R. and Holden, L.R. (1996) Insect bioassay for determining soil
degradation of Bacillus thuringiensis subsp. kurstaki CryIA(b) protein in
corn tissue. Environmental entomology, 25, 659-664.
Tabashnik, B.E., Carrière, Y., Dennehy, T.J., Morin, S., Sisterson, M.S., Roush,
R.T., Shelton, A.M. and Zhao, J.-Z. (2003) Insect resistance to transgenic
Bt crops: lessons from the laboratory and field. J. Econ. Entomol., 96, 10311038.
Tiedje, J.M., Asuming-Brempong, S., Nusslein, K., Marsh, T.L. and Flynn, S.J.
(1999) Opening the black box of soil microbial diversity. Appl. Soil Ecol.,
13, 109-122.
Torres, J.B. and Ruberson, J.R. (2005) Canopy- and ground-dwelling predatory
arthropods in commercial Bt and non-Bt cotton fields: patterns and
mechanisms. Environ. Entomol., 34, 1242-1256.
46
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Monsanto Company
Toth, F., Arpas, K., Szekeres, D., Kadar, F., Szentkiralyi, Szenasi, A. and Kiss, J.
(2004) Spider web survey or whole plant visual sampling? Impact
assessment of Bt corn on non-target predatory insects with two concurrent
methods. Environ. Biosafety Res., 3, 225-231.
US EPA. (2000) Bt-plant pesticides biopesticides registration document: section
C. Environmental Assessment. United States Environmental Protection
Agency.
US EPA. (2001) Biopesticides Registration Action Document: Bacillus
thuringiensis (Bt) Plant-incorporated Protectants. U.S. Environmental
Protection Agency,
http://www.epa.gov/pesticides/biopesticides/pips/bt_brad.htm.
US EPA. (2002) Memorandum. Transmittal of meeting minutes of the FIFRA
Scientific Advisory meeting held August 27-29, 2002. SAP meeting
minutes No. 2002-05. .
US EPA. (2005) Biopesticides Registration Action Document. Bacillus
thuringiensis Cry34Ab1 and Cry35Ab1 proteins and the genetic material
necessary for their production (plasmid insert PHP 17662) in event DAS59122-7 corn. , 42.
Van Mellaert, H., Van Rie, J., Hofmann, C. and Reynaerts, A. (1988) Insectidical
crystal proteins from Bacillus thuringiensis: mode of action and expression
in transgenic plants. Conference on biotechnology, Biological pesticides and
novel plant-pest resistance for insect pest management.
Specificity of Bacillus thuringiensis delta-endotoxins-importance of specific
receptors on thebrush border membrane of the mid-gut of target insect.
Eur. J. Biochem., 186, 239-247.
Receptors on the brush border membrane of the insect midgut as
determinants of the specificity of Bacillus thuringiensis Delta-endotoxins.
Applied and environmental microbiology, 1378-1385.
Vercesi, M.L., Krogh, P.H. and Holmstrup, M. (2006) Can Bacillus thuringiensis
(Bt) corn residues and Bt-corn plants affect life-history traits in the
earthworm Aporrectodea caliginosa? Applied Soil Ecology, 32, 180-187.
Vinson, S.B. (1989) Potential impact of microbial insecticides on beneficial
arthropods in the terrestrial environment. Safety of Microbial Insecticides,
43-64.
Vojtech, E., Meissle, M. and Poppy, G.M. (2005) Effects of Bt Maize on the
herbivore Spodoptera littoralis (Lepidoptera: Noctuidae) and the parasitoid
Cotesia marginiventris (Hymenoptera: Braconidae). Transgenic Research,
14, 133-144.
Volkmar, C. and Freier, B. (2003) Spinnenzoenosen in Bt-mais un nicht
gentechnisch veränderten Maisfeldern. Zeitschrift für Pflanzenkrankheiten
und Pflanzenschutz (J. Plant Diseases and Protection), 110, 572-582.
47
MON 810
Monsanto Company
Wandeler, H., Bahylova, J. and Nentwig, W. (2002) Consumption of two Bt and
six non-Bt corn varieties by the woodlouse Porcellio scaber. Basic and
Applied Ecology, 3, 357-365.
Weber, T.E., Richert, B.T., Kendall, D.C., Bowers, K.A. and Herr, C.T. (2000)
Grower-finisher performance and carcass characteristics of pigs fed
genetically modified "Bt" corn.
http://www.ansc.purdue.edu/swine/swineday/sday00/psd07-2000.html,
1-8.
West, A.W. (1984) Fate of the insecticidal, proteinaceous parasporal crystal of
Bacillus thuringiensis in soil. Soil Biol. Biochem, 16, 357-360.
West, A.W., Burges, H.D., White, R.J. and Wyborn, C.H. (1984) Persistence of
Bacillus thuringiensis parasporal crystal insecticidal activity in soil. J.
Invertebr. Pathol., 44, 128-133.
Whitehouse, M., Wilson, L. and Fitt, G. (2005) A comparison of arthropod
communities in transgenic Bt and conventional cotton in Australia.
Whiteley, H.R. and Schnepf, H.E. (1986) The molecular biology of parasporal
crystal body formation in Bacillus thuringiensis. Annu Rev Microbiol, 40,
549-76.
Wolfersberger, M.G., Hofmann, C. and Luthy, P. (1986) Interaction of Bacillus
thuringiensis delta-endotoxin with membrane versicles insolated from
lepidoteran larval midgut. Bacterial protein toxins, 237-238.
Yao, H., Ye, G., Jiang, C., Fan, L., Datta, K., Hu, C. and Datta, S.K. (2006) Effect
of the pollen of transgenic rice line, TT9-3 with a fused cry1Ab/cry1Ac gene
from Bacillus thuringiensis Berliner on non-target domestic silkworm,
Bombyx mori L. (Lepidoptera: Bombyxidae). Appl. Entomol. Zool., 41, 339348.
Zangerl, A.R., McKenna, D., Wraight, C.L., Carroll, M., Ficarello, P., Warner, R.
and Berenbaum, M.R. (2001) Effects of exposure to event 176 Bacillus
thuringiensis corn pollen on Monarch and black swallowtail caterpillars
under field conditions. Proc. Nat. Acad. Sci, 98, 11908-11912.
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MON 810 maize
May 2007
APPENDIX 2
EU WORKING GROUP ON
INSECT RESISTANCE MANAGEMENT
- Harmonised insect resistance management (IRM) plan
for cultivation of Bt maize in the EU -
January 13, 2003
Participating companies:
Monsanto Europe S.A.
Pioneer Hi-Bred International, Inc.
Syngenta Seeds S.A.S.
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EU WG IRM plan, January 13 2003
Table of contents
1
INTRODUCTION ................................................................................................................. 4
2
SCOPE OF THE PLAN........................................................................................................ 5
3
APPROACH AND RATIONALE OF THE PLAN............................................................ 5
3.1
THE EUROPEAN UNION .................................................................................................... 5
3.1.1
Current legislation addressing monitoring ............................................................. 5
3.1.2
European Commission Scientific Committee on Plants (SCP)................................ 6
3.1.3
Spain........................................................................................................................ 6
3.2
PRACTICAL EXPERIENCE WORLDWIDE ............................................................................ 7
3.2.1
USA.......................................................................................................................... 7
3.2.2
Argentina ................................................................................................................. 7
3.3
CURRENT INDUSTRY EFFORTS IN HARMONISATION OF MONITORING REQUIREMENTS
AND IRM...................................................................................................................................... 7
4
CHARACTERISTICS OF THE IRM PLAN ..................................................................... 8
4.1
4.2
5
EFFECTIVE ....................................................................................................................... 8
BALANCED AND PRACTICAL ............................................................................................ 8
ELEMENTS OF THE IRM PLAN...................................................................................... 9
5.1
REFUGE ............................................................................................................................ 9
5.1.1
Refuge size............................................................................................................... 9
5.1.2
Refuge configuration and placement..................................................................... 10
5.1.3
Refuge management............................................................................................... 10
5.2
RESISTANCE MONITORING ............................................................................................. 10
5.2.1
Objectives and underlying principles .................................................................... 10
5.2.2
Monitoring protocol .............................................................................................. 11
5.3
REMEDIAL PLAN IN CASE OF BT MAIZE FAILURE TO PROTECT AGAINST TARGET PESTS 13
5.3.1
Procedures for unexpected damage ...................................................................... 13
5.3.2
Steps to confirm resistance.................................................................................... 13
5.3.3
Remedial actions if insect resistance is confirmed................................................ 13
6
IMPLEMENTATION (GROWER EDUCATION) ......................................................... 14
7
REFERENCES .................................................................................................................... 15
8
ABBREVIATION/DEFINITION OF TECHNICAL TERMS........................................ 18
9
APPENDICES ..................................................................................................................... 19
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Summary
Maize is an important crop in the European Union (EU) and corn borer infestations can result in
considerable crop damage and yield loss. In Spain for example, the losses in maize production
can be as high as 15% in areas of high corn borer pressure. The use of conventional insecticides is
not practical against corn borers since chemical sprays cannot reach boring pest larvae whereas
crops expressing a Bacillus thuringiensis protein for pest resistance, hereafter referred to as Bt
crops, provide an in-built resistance to combat these pests. Since its introduction in the USA in
1995, Bt maize has proved to be a successful management tool to control crop damage and losses
to insect pests. This technology has been widely adopted and its use extended to other countries.
With the introduction of Bt crops, concerns have been raised about the possible development of
insect resistance that could deprive growers of the benefits of Bt crops and Bt microbial
preparations. Despite the fact that resistance to Bt in the field has not been detected to date, this
concern has been addressed pro-actively in a number of countries by the implementation of insect
resistance management plans to delay the potential development of pest resistance and to enable
the timely detection of changes in pest susceptibility. Following the experiences gained in other
countries and taking into account the latest scientific reports, an industry working group, the EU
Working Group on Insect Resistance Management has developed a harmonised insect
resistance management (IRM) plan specific for the EU.
The objectives of the Working Group are to assist in compliance with existing EU regulatory
requirements with regard to the monitoring of Bt maize and to protect the Bt technology through
good stewardship. The purpose of the IRM plan is to proactively avoid where possible, and in all
cases delay the potential development of pest resistance to the Cry1Ab and Cry1F proteins as
expressed in Bt maize. The Working Group is well aware that the key to success for the IRM plan
is acceptance and adoption by European growers. Therefore it has been designed to be effective,
balanced and practical. The harmonised IRM plan contains guidance on the following key
elements:
•
How to use the Bt technology: a comprehensive grower education programme will aid the
grower in understanding the importance of insect resistance management to preserve the
long-term efficacy of the Bt technology and in employing the required resistance
management tool of implementing a generous 20 % refuge for Bt maize planting areas larger
than 5 hectares
•
Resistance monitoring: baseline susceptibility of European corn borer (Ostrinia nubilalis) and
Mediterranean corn stalk borer (Sesamia nonagrioides) to Cry1Ab and Cry1F endotoxins of
B. thuringiensis in the EU will be measured and monitoring techniques are described to detect
changes relative to baseline susceptibility which could result in inadequate protection against
O. nubilalis and S. nonagrioides in the field
•
Potential development of resistance: confirmation of pest resistance and remedial action plan.
In summary, the harmonised IRM plan for the EU is designed to allow farmers to benefit from
growing Bt maize while pro-actively avoiding where possible, and in all cases delaying the
potential development of target pest resistance in the field.
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1
INTRODUCTION
Bacillus thuringiensis (Bt) is a Gram-positive bacterium capable of producing large crystal
protein inclusions that have insecticidal properties. The efficacy and specificity of Bt strains and
individual toxins produced by Bt isolates are such that a large number of insecticidal products
based on this bacterium and/or its toxins have been developed and sold commercially since the
late 1950’s. Historically, Bt has been considered a safe option for pest control and it has often
been the preferred pest control method in Integrated Pest Management (IPM) programmes.
Using modern biotechnology, the genes coding for specific Bt toxins were isolated in the 1980’s
and introduced into various crop plants to provide insect protection. Such insect-protected crops
now represent an important new management tool to control crop damage and losses due to insect
pests. In addition, the use of insect protected crops will provide important benefits to growers,
society, and the environment (McGaughey and Whalon, 1992; Gasser and Fraley, 1989; Gould,
1988; Nester et al., 2002).
Maize expressing a Bacillus thuringiensis protein for pest resistance, hereafter referred to as Bt
maize, was first registered (deregulated) in the USA in 1995. Currently, it is the second most
widely planted genetically modified (GM) crop, with approximately 5.9 million hectares
commercially grown worldwide in 2001, mainly in the USA, Canada, Argentina, and South
Africa. The introduction of Bt maize in the European Union (EU) is at an early stage. Small areas
of Bt maize, approximately 20 - 25,000 ha or 5 % of the total Spanish maize growing area, have
been planted each year in Spain since 1998. Token amounts have been planted in France in 2000
and in Germany in 2001 (ISAAA, 2001). A recent study (Brookes, 2002) demonstrated clear
economic and environmental benefits for Spain in the Bt maize growing areas.
Although to date there is no literature report of ECB developing resistance to Bt microbial
products in the field, the potential occurrence of resistance to Bt proteins arising from the use of
Bt crops is generally accepted as a possibility. Therefore, in the interests of good product
stewardship and to preserve the utility of the technology, the large-scale commercial introduction
of Bt maize in various world areas has been accompanied by insect resistance management (IRM)
plans. So far, no cases of insect resistance to Bt maize have been reported anywhere.
An EU Working Group on Insect Resistance Management was formed in late 2001 to proactively prepare for large-scale cultivation of Bt maize in the EU. The Working Group represents
an industry collaboration intending to harmonize the IRM plan for cultivation of Bt maize in the
EU and other relevant European markets. This group was set up in light of the positive experience
acquired in the USA with a comparable industry group, the Agricultural Biotechnology
Stewardship Technical Committee (ABSTC) IRM Subcommittee.
The objectives of the EU Working Group are to:
(a) Assist in compliance with existing regulatory requirements (EU Directive 2001/18/EC).
(b) Take the lead in determining the requirements for monitoring of Bt maize, recognizing
that the EU Commission has already considered some aspects of IRM such as developing
a draft protocol for the monitoring of European corn borer resistance to Bt maize (EC
Document XI/157/98).
(c) Protect Bt technology through good stewardship.
The Working Group currently involves the following industry partners: Monsanto Europe S.A.,
Pioneer Hi-Bred International, Inc. and Syngenta Seeds S.A.S.
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2
SCOPE OF THE PLAN
The purpose of this harmonised IRM plan is to pro-actively avoid where possible, and in all cases
delay the potential development of resistance to the Cry1Ab and Cry1F proteins, as expressed in
Bt maize. The harmonised IRM plan also includes timely detection of changes in pest
susceptibility and remedial actions in case of any confirmed development of resistance. The
transformation events currently included in the proposal are presented in Table 1.
Table 1: Proteins and transformation events currently included
in the harmonised IRM plan
Transformation
event
OECD unique
identifier
Protein
Notifier
Bt 176
SYN-EV176-9
Cry1Ab
Syngenta
Bt 11
SYN-BT∅11-1
Cry1Ab
Syngenta
MON 810
MON-∅∅81∅-6
Cry1Ab
Monsanto
1507
DAS-∅15∅7-1
Cry1F
Pioneer;
Mycogen/DAS
The main insects targeted by the plan are the European corn borer and the Mediterranean corn
stalk borer, as shown in Table 2.
Table 2: Insects targeted by the harmonised IRM plan
3
Common name
Abbrev.
Scientific name
Family
European corn borer
ECB
Ostrinia nubilalis (Hubner)
Crambidae
Mediterranean corn stalk borer
MCB
Sesamia nonagrioides (Lefebvre)
Noctuidae
APPROACH AND RATIONALE OF THE PLAN
The success of the harmonised IRM plan is highly dependent on grower acceptance and
implementation of the proposed management practices designed to preserve pest susceptibility to
Bt proteins. In developing the IRM plan, the Working Group considered the latest scientific
findings in order to both protect Bt technology and establish a practical, logistically achievable,
approach that growers can follow.
The harmonised IRM plan is therefore based on published research (cited throughout the
document), current EU legislation, the European Commission’s Scientific Committee on Plants
(SCP) opinion on IRM (SCP, 1999) and practical experience gained during the implementation of
IRM plans in other parts of the world.
3.1
3.1.1
The European Union
Current legislation addressing monitoring
Directive 2001/18/EC, Annex VII, requires that notifiers develop and submit a monitoring plan
together with the notification for placing on the market of a genetically modified (GM) crop. The
objectives of the monitoring plan are to confirm that any assumptions made regarding the
occurrence and impact of potential adverse effects of the GMO or its use in the environmental
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risk assessment (e.r.a.) are correct and to identify 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 European
Commission Scientific Committee on Plants (SCP).
3.1.2
European Commission Scientific Committee on Plants (SCP)
The European Commission’s Scientific Committee on Plants (SCP, formerly known as Scientific
Committee on Pesticides) has given a number of opinions on the cultivation of Bt maize in the
EU. In their opinion of 09 December 1996 on the use of GM maize lines notified by Ciba-Geigy,
the SCP stated that “Possible development of insect resistance to the Bt-toxin cannot be
considered an adverse environmental effect, as existing agricultural means of controlling such
resistant species of insects will still be available” (SCP, 1996).
In addition, the SCP has been consulted on the insect resistance monitoring program. After the
first EU approval for Bt maize cultivation in 1997 (Bt 176), the European Commission’s
Directorate General for Environment (DG ENV) established an ad-hoc expert group consisting of
representatives from EU Member States to develop a protocol to monitor the development of
resistance to Bt proteins in target pests of maize. The ‘Draft protocol for the monitoring of
European corn borer resistance to Bt maize (EC Document XI/157/98)’ was then forwarded to the
SCP for a scientific opinion. The SCP opinion was expressed on 4 March 1999 with the following
conclusions (SCP, 1999):
•
The draft protocol to monitor for resistance in European corn borer to Bt maize should be
linked firmly to field management plans on the ground. The proposed protocol, based on
experience and practical data, should be broadened by the inclusion of more sensitive
laboratory tests to detect low frequency resistance alleles and should include a suitable
discriminating dose. Monitoring should be targeted to more extensive areas of planted Bt
maize where selection pressures may be highest for resistance development and
procedures adjusted according to the GM variety grown (in relation to seasonal decline of
toxin concentration).
•
Effective high-dose structured refuge resistance management plans should be
implemented for Bt maize. Initially very large refuges may exist in practice, as the total
area of Bt maize will be small in Europe. However, plans should also take account of
spatial structuring on a local scale.
•
Growers should be required to survey their growing Bt maize to detect unusual patterns
of pest damage and ineffective control. Companies should thoroughly investigate these
incidents by insect sampling and laboratory testing. Investigations should include the
collection of plant material and the laboratory determination of expression levels in
tissue, and the assessment of pest densities in the Bt crop and surrounding vegetation at
the time of the problem.
3.1.3
Spain
GM maize expressing Cry1Ab protein (Bt 176) has been commercially grown in Spain since
1998 on approximately 20 – 25,000 ha annually. A detailed IRM plan was submitted to the hybrid
registration authorities asking growers to implement the proposed IRM measures.
Commercialisation of Bt 176 was accompanied by a monitoring research project to ensure early
detection of European corn borer resistance through regular monitoring on Bt maize fields.
Baseline susceptibility to the Cry1Ab toxin was also determined for Spanish populations of the
ECB and MCB collected on non-Bt maize (Gonzalez-Nuñez et al., 2000; De la Pozza et al.,
2001).
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3.2
3.2.1
Practical experience worldwide
USA
With the registration of the first Bt maize in the USA in 1995 (Bt 176), it was foreseen that target
pests increasingly exposed to Bt maize might develop resistance to Cry proteins. Measures such
as the implementation of refuge zones constituted by non-Bt maize were envisioned to preserve
the utility of the Bt technology. While prior to commercialisation an optimal refuge size and
structure could not be determined by the registrants in the USA, it was thought that the market
penetration of these crops would be sufficiently slow that considerable non-Bt maize remained to
act as natural refuges as further research was conducted. Various voluntary refuge requirements
were recommended. Efforts by both academia and industry were undertaken to develop a
harmonised, science-based and practical approach to IRM. In 1997, a report was published on ‘Bt
corn and European corn borer’ (Ostlie et al., 1997)) and a resistance management plan for
European corn borer were proposed (record where Bt maize is planted, implement 20-30 %
refuge of non-Bt maize, monitor for product failure). In 1998, the United States Environmental
Protection Agency (US EPA) began to institute refuge requirements for Bt maize.
In 1999, the industry ABSTC IRM Subcommittee, working with the National Corn Growers
Association (NCGA) and in consultation with academia and the US EPA, proposed a harmonised
IRM plan for Bt maize cultivation. With some modifications to this plan, US EPA put in place a
consistent set of required refuge strategies for all Bt maize products. Beginning with the growing
season in 2000, US EPA required a 20% non-Bt maize refuge to be planted within a half a mile of
the Bt maize field (US EPA, 2001).
3.2.2
Argentina
Approval of the first Bt maize line in Argentina occurred in 1997 (Bt 176), followed by
MON 810 in 1998. These products were introduced with a variety of voluntary IRM practices. In
1999, building upon the experiences in the USA, consultations began in Argentina among the Bt
maize registrants on the possibility of developing a joint industry IRM plan. These consultations
were held under the aegis of the Argentine Seed Association (ASA) and included third party
entomologists from academia and the Instituto Nacional de Tecnología Agropecuaria (INTA).
This group proposed a harmonised plan with a 10% refuge requirement. The basis for this plan
included knowledge of pest biology and grower behaviour. In particular, it was noted that the
presence of abundant alternative hosts for the target pests justified refuge sizes smaller than in the
USA for target pests that were otherwise similar in their biology. The IRM plan also included the
development of baseline susceptibility measurements for the target pests, the creation of
standardised educational literature for growers and the use of regular surveys to assess grower
compliance with the requirements. The joint industry IRM plan was accepted by the regulatory
agency Comisión Nacional Asesora de Biotecnología Agropecuaria (CONABIA) and
implemented.
3.3
Current industry efforts in harmonisation of monitoring requirements and IRM
The Technical Advisory Group (TAG) of EuropaBio’s Plant Biotechnology Unit has developed a
series of documents including monitoring of GM crops, to assist with harmonization regarding
the technical information submitted to the EU regulatory authorities via applications for the
commercial approval of GM crop products. Document 3.1 addresses the general approach to
monitoring, and Document 3.2. discusses more specifically monitoring of insect-protected Bt
crops (EuropaBio, 2001). In the case of existing Bt maize products, no adverse effects requiring
case-specific monitoring have been identified in the environmental risk assessment (this
conclusion is in agreement with the above mentioned SCP opinion (SCP, 1996)). However, to
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ensure good product stewardship, an Insect Resistance Management Plan for Bt maize is
considered as critical by the industry. Therefore, since 2001, the work initiated by the TAG group
has been specifically followed up by establishing the EU Working Group on Insect Resistance
Management involving relevant industry partners.
4
CHARACTERISTICS OF THE IRM PLAN
For an IRM plan to be successful it must not only delay any development of pest resistance to Bt
maize but also it must be effective, balanced and practical for the users such as growers of Bt
maize.
4.1
Effective
Based on current knowledge of pest biology and insect resistance, combined with information
from simulation models incorporating highly generous safeguard margins, a science based IRM
plan has been developed.
Recognising that available data may not be representative of all pest populations and that a degree
of uncertainty exists, the present IRM plan incorporates generous safeguard margins to ensure
that the IRM plan is precautionary. In particular, the added safeguard margins are manifested by a
larger refuge than would be necessary in the EU on strictly technical grounds. A comparable
refuge strategy has been used in the USA where Bt maize has been grown widely on a
commercial scale since 1996. Despite extensive monitoring efforts over the past 6 years, there has
been no report of development of ECB resistance to Bt maize in the USA (Siegfried and Spencer,
2002).
The effectiveness of the IRM plan will be reviewed regularly, taking into account the results of
resistance monitoring to incorporate any new scientific developments relevant to the IRM plan.
4.2
Balanced and practical
It is important that all stakeholders of Bt maize technology adopt and implement the elements of
the IRM plan. Seed companies have experience in cooperating with regulatory agencies,
providing grower education, implementing product stewardship and working with experts on
resistance management initiatives. However, farming practices are also critical to the success of
the IRM plan. This highlights the importance of the decision-making of individual growers in the
implementation of the IRM plan, in particular the refuge strategy. These important factors have
been taken into consideration whilst developing the IRM plan, in particular the recommendations
for implementation of a refuge, which have been carefully designed to be pragmatic, clear and
consistent across relevant regions as well as provide a degree of flexibility where necessary
according to variable cropping systems.
The refuge requirement is part of the IRM plan and will delay the potential development of
resistance by target pests to Bt maize. This is a precautionary measure to reduce the selective
pressure on local populations of target pests. Details on refuge size, location, configuration and a
tested process for investigating unexpected damage are provided in the IRM plan. The practices
described in this plan balance a grower’s opportunity to benefit from Bt maize in the short term
with the longer-term objective of preserving the efficacy of Bt maize. All companies subscribing
to the present Working Group are committed to provide farmers with the necessary guidance,
technical support and advice on best practices for growing Bt maize.
- Page 8/28 -
5
ELEMENTS OF THE IRM PLAN
The IRM plan is comprised of four elements:
•
Maintaining an adequate level of non-Bt maize refuge in the vicinity of Bt maize to support
a sufficient local population of susceptible target pests.
•
Monitoring for any potential development of resistance.
•
Remedial action plan in case of any confirmed development of resistance.
•
Programme of grower education for greater awareness of Bt maize cultivation and proper
stewardship
The first three elements are elaborated below. Details about grower education can be found in
Section 6.
5.1
Refuge
Currently it is widely accepted that resistance to Bt crops is rare and genetically recessive. This
has led to the development of IRM plans using an effective dose plus refuge strategy based on the
following assumptions:
•
Bt plants express high levels of Bt toxin
•
Resistance alleles typically are partially or fully recessive and rare so there will be
few homozygous survivors
•
Refuges are set up so that resistant homozygotes will mate randomly with susceptible
homozygotes that do not survive the Bt crop.
In summary, the purpose of the refuge is to maintain high numbers of susceptible homozygotes
that will breed with the few surviving heterozygotes as well as with the rare resistant
homozygotes, thereby delaying the evolution of resistance.
The effectiveness of a refuge is dependent on biological, genetic, behavioural and social or
cultural factors. Therefore, the refuge strategy described below takes into account EU target pests,
agronomic conditions and cultural practices. Moreover, it draws from experience gained through
several years of implementing refuge strategies in countries where Bt maize is routinely
cultivated. The result is a refuge strategy that incorporates generous safeguard margins and will
avoid where possible, and in all cases delay resistance of target pests to Bt maize without
compromising grower accessibility to Bt maize or grower ability to implement refuge
requirements.
5.1.1
Refuge size
An appropriate level of refuge should be determined based on a comparative analysis of refuge
strategies and maize-growing conditions in countries where Bt maize is regularly cultivated. The
minimum proportion of non-Bt refuge implemented in the USA and Argentina is 20% and 10%,
respectively. Such refuge sizes are considered to contain generous safeguard margins under the
respective growing conditions, as described in Sections 3.2.1 and 3.2.2, respectively. A
comparative analysis between agricultural landscapes in the USA and the EU highlights the
fragmented and diverse cropping conditions in the EU. This explains why the current refuge
requirements in the USA of 20% is considered highly generous for the EU (Appendix 1), thereby
providing justification for a potentially lower level of refuge for the EU.
For the purpose of the present harmonised IRM plan for the EU, a grower is defined as the
individual responsible for seed purchasing and planting decisions on one farm. Growers planting
- Page 9/28 -
more than 5 hectares (ha) of Bt maize would be required to plant a non-Bt maize refuge whereas
growers planting less than 5 ha of Bt maize would not. This 5 ha threshold relates to the total
amount of Bt maize, within or among fields, planted by one grower and is independent of the size
of the individual fields or the total land area managed by this grower. As a consequence, the
requirement for refuge can only be applicable to farm sizes of more than 5 ha. The logistical
barriers to implementing an effective refuge on small fields or farms and the reasons why the 5 ha
threshold will not pose a resistance risk are outlined in Appendix 2.
5.1.2
Refuge configuration and placement
Refuge maize can be located near, adjacent to or within Bt maize fields. Refuges within a Bt
maize field can be planted as a block, perimeter border or strips (Appendix 3). Growers should
also ensure that the refuge maize and the Bt maize share similar growth and development
characteristics.
Growers should plant the refuge within 750 meters of their Bt maize field(s) although lesser
distances are preferred. The objective of this distance requirement is to maintain a high
probability of pest immigration into Bt maize, and consequently, a high probability that any rare
individuals surviving on Bt maize will mate with susceptible individuals from the refuge. The
scientific basis for this distance requirement is outlined in the work of Showers et al. 2001 and
Hunt et al. 2001, and this distance is consistent with refuge strategies practiced in other countries.
Guidelines for planting a refuge will be clearly communicated in the product use guide that
accompanies Bt maize (see example in Appendix 3). Additional educational materials are
addressed in Section 6 (see also NCGA point of purchase pamphlet in the reference list).
5.1.3
Refuge management
Refuge zones should be managed in the same way as the Bt crop areas, where possible. Growers
are encouraged to monitor their maize crop and control pest populations in non-Bt refuge maize
only when the level of pest damage reaches economic importance. Where necessary, insecticides
should be used according to their label recommendations. Microbial Bt sprays are the only class
of insecticide that must not be used in refuge maize.
5.2
Resistance monitoring
5.2.1
Objectives and underlying principles
The objectives of the resistance monitoring programme are to:
•
Measure the baseline susceptibility of European corn borer (Ostrinia nubilalis, Hubner)
and Mediterranean corn stalk borer (Sesamia nonagrioides, Lefebvre) to Cry1Ab (var.
kurstaki) and Cry1F (var. aizawai) endotoxins of B. thuringiensis.
•
Detect changes relative to baseline susceptibility that could result in inadequate
protection against O. nubilalis and S. nonagrioides in the field.
The establishment of baseline susceptibility measurements for Bt maize will be an extensive and
discrete effort. Subsequent monitoring for insect resistance will be an ongoing function that
should be flexible and account for the factors that may result in the development of pest
resistance to Bt maize.
In order to be effective, resistance monitoring should focus on areas of high selection for
resistance, including those with the highest intensity of Bt maize. A regional or country-bycountry component may be necessary; the former reflects biological reality and efficiency, and
the latter the special needs of specific countries cultivating Bt maize.
- Page 10/28 -
It should be recognized that technical, biological and practical limitations exist. However, once
baseline susceptibility is established, a monitoring programme designed as an integral part of the
IRM plan, should provide the flexibility to increase monitoring in a certain area if the need arises.
The extent and intensity of a resistance monitoring programme should therefore be balanced
against other components of the IRM plan, such as specific efforts to survey and promote the
implementation of IRM measures (e.g. refuge), or the monitoring of product adoption. For
example, insect resistance monitoring could become more important if adoption of Bt maize is
relatively high or if specific technical or practical challenges predict difficulties in reaching
favourable levels of refuge implementation. The laboratory-based resistance monitoring efforts
will be supported by a programme to follow product performance and unexpected damage.
5.2.2
Monitoring protocol
5.2.2a Baseline susceptibility
A number of studies have already been performed to measure baseline susceptibility (Marćon et
al., 1999 and 2000; Gonzalez-Nuñez et al., 2000 and Wu et al., 2002). These have been
conducted using different sources of protein and slightly varying methods. To reduce the level of
variability and allow true data comparisons, it is important to perform the baseline susceptibility
measurements for the EU in a coordinated manner using common sampling methods and sources
of protein. A thorough report should accurately describe and contrast the baseline susceptibility of
each individual population sampled. Any subsequent effort should follow the established methods
and normalise the results obtained.
There are two general objectives of a baseline susceptibility study:
•
To measure the susceptibility of target pest populations that may be exposed to Cry1Ab
and Cry1F proteins in Bt maize.
•
To measure the variability in susceptibility to Cry1Ab and Cry1F proteins that exists
between ECB and MCB populations.
The former objective is used to determine the sensitivity of individual pest populations across
geographic regions. The latter can be used to gauge the importance of potential shifts in
susceptibility that may be detected in subsequent resistance monitoring efforts. A successful
study of baseline susceptibility should carefully weigh the precision of estimates for individual
populations against the scope or number of populations included in the baseline study.
The design of the baseline susceptibility study is a function of the factors that may increase the
rate at which target pest species adapt to Bt maize. These factors are defined in insect biology and
ecology, and will be linked to the distribution and intensity of Bt maize production throughout the
EU. A summary of the important factors to be considered prior to the baseline susceptibility study
is given in Appendix 4. Consideration of these factors suggests possibly four focussed regions
based on where Bt maize cultivation could be greatest and where the intensity of target pest
infestation could be relatively high.
The baseline study should determine the susceptibility of populations of target pests from three
locations within each focus regions (e.g. a total of 12 populations for each target pest from the
four focus regions). Additions or substitutions to these locations can be a function of target pest
occurrence and feasibility of collection.
Participating companies will coordinate the insect collections. Collection efforts should focus on
the egg stage for efficiency; however, any stage of insect may be collected to reach the required
number of populations for each target pest.
- Page 11/28 -
Assays used to determine baseline susceptibility should be performed on F1 progeny whenever
possible, and 200 to 300 insects should be collected from each sample location (population).
Sample locations should be omitted if insect collections produce less than 100 individuals.
As recommended by the SCP (SCP, 1999), diet bioassays using F1 insects should be used to
develop a 7-day LC50 and EC50, and subsequently an LC99 and EC99 or a discriminating dose for
use in future resistance monitoring. The methodology used in these assays should follow the
methods described in the published work of Marćon et al. (1999, 2000) and Gonzalez-Nuñez et
al. (2000). Specifically, the study should use seven to nine concentrations of each Cry protein
(supplied by the Working Group) in a diet-overlay format. Estimates for each concentration
should be based on no less than 60 total individuals using an appropriate experimental design
with replication. When different events are expressing the same insecticidal activity, the baseline
susceptibility measurements can be performed and applied to the different events using the same
protein.
5.2.2b Resistance monitoring
Monitoring should be focused in areas where adaptation of target pests to Bt maize is more likely
due to relatively high Bt maize cultivation in line with high target pest infestation.
The monitoring outlined below is designed to detect resistance when the frequency of the
resistant allele reaches about 1-5%. For the proposed collection sizes, the upper 95% confidence
limit for a dominant or partially dominant allele is 0.8%, while the corresponding 95% confidence
limit for a recessive allele is 9%. However, these methods may not detect very small shifts in
resistance allele frequency (ABSTC, 2002). Nonetheless, this level of sensitivity should allow for
early detection of potential pest resistance before field failures occur and therefore would enable
additional management measures to be effectively implemented in a timely manner.
Data generated during the baseline susceptibility study should be used to develop a technically
accurate and more cost effective assay capable of detecting statistically and biologically
significant changes in susceptibility within a population. A discriminating dose assay fits this
goal, and the practicality of developing this assay should be evaluated after the baseline studies
are complete (Hawthorn et al., 2001). Marćon et al. 2000 provides useful background on the
discriminating dose assays.
As discussed earlier, the extent and intensity of resistance monitoring will be a function of the
extent of Bt maize adoption and the intensity of target pest infestation. Geographically referenced
measures of adoption will be performed as necessary and these measures will be inclusive of all
Bt maize containing Cry1Ab or Cry1F and independent of company or brand name. Measures of
adoption will be made at a resolution much smaller than the likely extent of the focus regions.
This should increase the efficiency of the resistance monitoring programme. The analysis of
adoption will be carried out by a third party at the expense of the Working Group. Results will be
communicated to the Working Group and used to determine the necessary sampling efforts.
Insect collections will be coordinated by participating companies. Assays used to determine
baseline susceptibility should be performed on F1 progeny whenever possible, and sample
locations should be omitted if insect collections produce less than 100 individuals.
Tests for significant deviations from baseline LC50 and EC50 should be used for resistance
monitoring until a discriminating dose assay is developed. The laboratory procedures for these
LC50 and EC50 assays should be the same as those used in the measurement of baseline
susceptibility.
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5.3
5.3.1
Remedial plan in case of Bt maize failure to protect against target pests
Procedures for unexpected damage
The following procedures are proposed in case of unexpected damage:
a. The seed company will require distributors to instruct purchasers of Bt maize seed to
report unexpected levels of damage caused by target pests.
b. The seed company will provide distributors specified information, including details
of the report, grower contact details.
c. If the company is a licensee for the Bt trait, it will transmit this information to the
notifier.
d. Companies will investigate the cause of these reports using available methods to
confirm that the damaged plants express Cry protein, the damage resulted from a
target pest and the damage is unexpected.
e. Insects will be collected for the purpose of further evaluation and confirmation
subject to subsequent investigations to confirm that damage is unexpected.
5.3.2
Steps to confirm resistance
a. If damage is unexpected, the collected insects will be tested in a laboratory following
specific guidelines used to confirm resistance.
b. Both of the following conditions must be met to confirm resistance: the collected
insects or their progeny must have an LC50 that exceeds the upper 95% confidence
interval of the historical (susceptible) mean LC50 for the appropriate Bt protein and
the collected insects or their progeny must achieve > 30% survival and >25% leaf
area damage in a 5-day bioassay under laboratory conditions using the appropriate
protein-positive leaf tissue.
5.3.3
Remedial actions if insect resistance is confirmed
a. A remedial action plan will be developed, involving the relevant notifier and others
concerned with the cultivation of affected Bt maize in collaboration with the
pertinent Member State authority.
b. Components of an appropriate remedial action plan may include:
-
Informing customers and extension agent in the affected areas of confirmed
resistance.
-
Increasing monitoring in affected areas.
-
Implementing alternative means to reduce or control target pest populations
in affected areas.
-
Modifying and amending the IRM strategy accordingly.
-
If the above measures are not efficient, then cessation of sales in the affected
and bordering areas may be necessary until an effective local management
plan approved by the pertinent Member State has been put in place.
If interrupted, sale of Bt maize in the affected area will restart when an effective
management plan has been implemented.
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6
IMPLEMENTATION (GROWER EDUCATION)
An extensive grower education programme is essential for the successful implementation of the
IRM plan. Growers should have a clear understanding of the importance of IRM to preserve the
long-term efficacy of the Bt technology and realise that their participation in this IRM
stewardship programme is vital to prolonging the success and benefits of Bt maize. Each of the
seed companies participating in this IRM plan is committed to continuing with their ongoing
comprehensive education programmes.
A technical user guide will provide each purchaser of Bt maize with latest information on the
recommendations for the IRM plan, Bt technology, the approval status of various Bt maize
hybrids in the relevant country and contact details of the responsible seed provider (technology
provider, licensee). The user guide will request growers to implement the required IRM measures
such as recording where Bt maize is planted, planting a non-Bt maize refuge and monitoring
product performance.
In addition, the IRM plan will be communicated using a combination of the following means:
•
Slide and video presentations to growers and distributors, co-ops, seed dealers and
distributors.
•
Information via company and relevant country specific association as well as agricultural
extension services web sites.
•
Newsletters.
•
Country specific hotlines.
•
Relevant competent authorities.
A common outline of the IRM guidance is provided in Appendix 3 and will be adapted to
the conditions of the local market.
- Page 14/28 -
7
REFERENCES
ABSTC (2002). Industry Advisory Panel to Agricultural Biotechnology Stewardship Technical
Committee (ABSTC) on Monitoring for Insect Resistance to Bt corn. Report submitted to
EPA.
Brookes G. (2002). The farm level impact of using Bt maize in Spain.
http://www.europabio.org/pages/ ne_gbgmcrops.asp
Caprio M.A., Luttrell R.G., MacIntosh S., Rice M.E., Siegfried B., Witkowski J.F., Van Duyn J.,
Moellenbeck D., Sachs E., and Stein J. An evaluation of insect resistance management in Bt
field corn: A science-based framework for risk assessment and risk management. ILSI Press:
Washington, DC. 85pp.
Cordero A., Malvar R.A., Butron A., Revilla P., Velasco P., and Ordas A. (1998). Population
dynamics and life cycle of corn borers in South Atlantic European coast. Maydica 43(1):
5 - 12.
De la Poza M., Farinós G.P., Hernández-Crespo P., Ortego F. and Castaňera P. (2001).
Monitoring of corn borer resistance to Bt-maize in Spain: forecast of resistance. XXI IWGO
Conference, Legnaro, Italy, Oct. 27 – Nov. 3, 2001 (abstract). Published in IWGO Newsletter
2002 / XXIII 1, p.9.
EuropaBio, TAG Working Group (2001). Safety assessment of GM crops. Document 3.1:
monitoring. http://www.europabio.org/pages/eu_workgroups_detail.asp?wo_id=14.
EuropaBio, TAG Working Group (2001). Safety assessment of GM crops. Document 3.2:
monitoring of insect-resistant Bt crops.
http://www.europabio.org/pages/eu_workgroups_detail.asp?wo_id=14.
European Commission (1998). Draft protocol for the monitoring of European corn borer
resistance to Bt maize. Document XI/157/98.
European Commission (1990). Directive 90/220/EC on the deliberate release into the
environment of genetically modified organisms. Official Journal of the European
Communities L117/15.
European Commission (2001). Directive 2001/18/EC on the deliberate release into the
environment of genetically modified organisms and repealing Council Directive 90/220/EC.
Official Journal of the European Communities L106: 1 – 38.
Gasser C.S. and Fraley R.T. (1989). Genetically engineering plants for crop improvement.
Science 244(4910): 1293-9.
Gonzalez-Nuñez M.G., Ortega F. and Castañera P. (2000). Susceptibility of Spanish populations
of the corn borers Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis
(Lepidoptera: Crambidae) to a Bacillus thuringiensis endotoxin. J. Econ. Entomol. 93(2):
459 - 463.
Gould F. (1998). Sustainability of transgenic insecticidal cultivars: Integrating pest genetics and
ecology. Annu. Rev. Entomol. 43:701-726.
Hawthorn D., Siegfried B., Shelton T. and Hellmich R. (2001). Monitoring for resistance alleles:
A report from an advisory panel on insect resistance monitoring methods for Bt corn. Report
submitted to EPA.
- Page 15/28 -
Hellmich, R.L., Pingel R.L. and Hansen W.R. (1998). Influencing European corn borer
(Lepidoptera: Crambidae) aggregation sites in small grain crops. Env. Entomol. 27(2): 253 259.
Hodgeson B.E. (1928). The host plants of the European corn borer in New England. US Dept.
Agr. Bul. 77.
Hunt T.E., Higley L.G., Witkowski J.F., Young L.J. and Hellmich R.L. (2001). Dispersal of adult
European corn borer (Lepidoptera: Crambidae) within and proximal to irrigated and nonirrigated corn. J. Econ. Entomol. 94(6): 1369 - 1377.
ISAAA (2001). Preview: global review of commercialised transgenic crops: 2001. ISAAA Briefs
no. 24 – 2001.
Losey, J.E., Calvin D. C., Carter M.E. and Mason C.E. (2001). Evaluation of non-corn host plants
as a refuge in a resistance management programme for European corn borer (Lepidoptera:
Crambidae) on Bt-corn. Environ. Entomol. 30(4): 728 - 735.
Marćon P.C.R.G., Young L.J., Steffey K.L. and Siegfried B.D. (1999). Baseline susceptibility of
European corn borer (Lepidoptera, Crambidae) to Bacillus thuringiensis toxins. J. Econ.
Entomol. 92(2): 279 - 285.
Marćon P.C.R.G., Siegfried B.D., Spencer T. and Hutchinson W.D. (2000). Development of
diagnostic concentrations for monitoring Bacillus thuringiensis resistance in European Corn
Borer (Lepidoptera, Crambidae). J. Econ. Entomol. 93(3): 925 – 930.
McGaughey, W.H. and M.E. Whalon (1992). Managing insect resistance to Bacillus thuringiensis
toxins. Science 258:1451-1455.
Nester E.W, Thomashow L.S., Metz M. and Gordon M. (2002). 100 years of Bacillus
thuringiensis: a critical scientific assessment. Washington DC: American Academy of
Microbiology. http://www.asmusa.org.
NCGA point of purchase pamphlet.
http://www.ncga.com/biotechnology/insectMgmtPlan/importance_bt.htm.
Ostlie K.R., Hutchinson W.D., and Hellmich R.L. (1997). Bt corn and European corn borer. NRC
publication 602. University of Minnesota, St. Paul.
http://www.extension.umn.edu/distribution/cropsystems/DC7055.html
SCP (1996). Opinion of the Scientific Committee for Pesticides on the use of genetically
modified maize lines notified by Ciba-Geigy (opinion expressed on 09 December 1996).
SCP (1999). Opinion of the Scientific Committee on Plants on Bt Resistance monitoring (opinion
expressed on 04 March 1999). Document SCP/GMO/094-Rev. 5.
Showers W.B., Hellmich R.L., Ellison D.R.M and Hendrix W.H. (2001). Aggregation and
dispersal behaviour of marked and released European corn borer (Lepidoptera: Crambidae)
adults. Env. Entomol. 30(4): 700 - 710.
Siegfried B. and Spencer T. (2002). Monitoring Bt susceptibility of European Corn Borer to
Cry1Ab; 2001 Data summary, Agricultural Biotechnology Stewardship Technical Committee
(ABSTC). Unpublished submission to EPA, April 30, 2002.
US EPA (2001) Biopesticides registration action document for Bacillus thuringiensis plantincorporated protectants (October 15, 2001)
http://www.epa.gov/pesticides/biopesticides/reds/brad_bt_pip2.htm.
- Page 16/28 -
Wu K., Guo Y., LV N., Greenplate J.T. and Deaton R. (2002). Resistance monitoring in
Helicoverpa armigera (Lepidoptera: Noctuidae) to Bacillus thuringiensis insecticidal protein
in China. J. Econ. Entomol. 95(4): 826 – 831.
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8
ABBREVIATION/DEFINITION OF TECHNICAL TERMS
Bt
Bacillus thuringiensis
Bt maize
Maize plants expressing Bt Cry proteins
Cry protein
Crystal protein derived from Bt
EC50
Effective concentration: the concentration, which affects 50% of a test
population after a specified exposure time
EC99
Effective concentration: the concentration, which affects 99% of a test
population after a specified exposure time
ECB
European corn borer
Endotoxin
Toxic molecule associated with the outer membrane and cell wall of
bacteria
GM
Genetic modification
Grower
Individual responsible for seed purchasing and planting
IPM
Integrated pest management
IRM
Insect resistance management
LC50
The median lethal concentration (i.e. the concentration/dose of substance
that is estimated to be lethal to 50% of the test organisms)
LC99
Lethal concentration (i.e. the concentration/dose of substance that is
estimated to be lethal to 99 % of the test organisms)
MCB
Mediterranean corn stalk borer
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9
APPENDICES
APPENDIX 1: Comparative analysis of EU and USA agricultural landscapes
Summary
A comparison of the landscape in the USA and the EU indicates that the adoption of a refuge
strategy based on the USA model of 20% non-Bt maize refuge in Europe would result in an
approach that incorporated highly generous safeguard margins for the harmonised IRM plan.
Paradigm
To base an IRM plan on the worst-case conditions with the highest probability to favour pest
resistance to Bt crops will incorporate generous safeguard margins. These conditions correspond
to where maize cultivation and Bt maize adoption are greatest and insect pressure is highest. In
the USA, this occurs in Nebraska, Iowa, Minnesota, Illinois and Indiana, commonly referred to as
the US Corn Belt. These five states routinely account for approximately 65% of the USA maize
crop area and represent the conditions with the highest potential for the development of insect
resistance. Conditions with highest probability of Bt maize adoption, and thus of potential
resistance development, in the EU are limited to France, Spain, Germany and Italy, where
approximately 85% of the EU maize is cultivated.
Comparisons of EU and USA agricultural landscapes
A comparison of these intensive maize-growing areas in the EU and the USA indicates that there
are important differences between them that make the risk of resistance development significantly
less for the EU than for the US Corn Belt. Examination of three key variables - land committed to
agriculture, farm size and crop diversity – clearly demonstrates that the EU agricultural landscape
is much more fragmented than that of the USA, thereby favouring greater durability of Bt maize
in the EU.
-
The US Corn Belt has an overwhelming 57 – 94% of its land committed to
agriculture, whereas the four key maize-growing EU countries have between 49
and 60% of their land committed to agriculture (Table 1).
-
France, Germany, Italy and Spain have an average of 10 to 20 times more farms
per unit of farmland compared to the US Corn Belt (Table 1). A greater number
of farms will result in increased crop diversity in an area.
-
Maize is the major crop of the US Corn Belt and constitutes 18 to 40% of the
total agricultural land and only slightly less of the total land area. However,
maize constitutes only 2 to 11% of the total agricultural land of EU Member
States. Moreover, non-maize cereal crops cover approximately 42%, 45% and
31% of the arable land in Germany Spain and France, respectively, and other
important crops such as sunflower, potato and sugar beet also serve as alternate
hosts for O. nubilalis (Hodgeson, 1928) (Table 2).
Conclusion
These data highlight the major differences between the EU and the USA maize-growing regions.
These contrasting differences in farming practices, resulting in a much lower potential of risk
resistance development, would suggest that an appropriate proportion of non-Bt maize refuge for
the EU could be less than the 20% currently used in the US Corn Belt.
- Page 19/28 -
Table 1: Comparison of farm numbers and commitment
of agricultural land for leading maize producing areas of the EU and the USA.
Number of farms
(x 1,000)
Total land area
(1,000 ha)
% of land in
agriculture
France
735
54909
55
Italy
2482
30132
56
Spain
1278
50488
60
Germany
567
35697
49
Nebraska
55
19911
94
Iowa
96
14472
92
Minnesota
80
20621
57
Illinois
79
14399
78
Indiana
65
9290
68
Europe:
USA:
Sources : Eurostat-Newcronos (1996) and USDA, NASS (2000)
Table 2: Summary of crop diversity for major maize-producing areas of the EU and the USA.
Agricultural Crop Land (1,000 ha)
Total
Ag.
Land
Area
Corn
(all)
Beans,
Peas,
Soy
Oats
Wheat
(all) Hay (all) Barley
Oilseed
Rape, Sunflow Sugar
Canola
er
Beet
Rye
France
30,060
3,307
2,786
170
5,115
.
1,534
41
1,369
799
Spain
30,126
545
83
409
2,423
.
3,107
122
48
850
Germany
17,344
1,698
706
309
2,601
.
2,210
748
1,198
33
Italy
16,743
1,314
349
142
2,388
.
350
5
51
209
Nebraska
18,785
3,439
1,902
53
708
1,254
Iowa
13,360
4,856
4,322
97
7
688
Minnesota
11,660
2,873
2,832
162
818
951
Illinois
Indiana
11,215
6,275
4,471
2,306
4,228
2,306
22
16
372
223
343
271
4
.
18
.
109
.
.
.
12
16
8
461
.
515
.
38
.
.
105
.
.
* Percent of total agricultural land area accounted for using these major and minor crops.
Sources: Eurostat 1995, 1999, USDA NASS 1999, 2000
- Page 20/28 -
32
196
.
.
Fresh
Veg.
Grape
Citrus
Olives
Fruit
Trees
171
321
873
3
13
172
136
388
1,166
288
2,350
979
309
90
101
86
.
32
.
.
Potato
.
.
908
.
182
1,154
55
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
2
.
APPENDIX 2: Using an area threshold to achieve an effective and practical refuge
Summary
The proposed refuge strategy for cultivation of Bt maize could consist of implementing a
generous 20% refuge on farms where a Bt maize area greater than 5 hectares (ha) is cultivated.
This strategy integrates the safeguards of a 20% refuge strategy in the European agricultural
landscape (see Appendix 1) and the practical convenience to smaller scale maize farmers. More
importantly, it also allows both large and small farmers to benefit from the cultivation of Bt
maize without adversely affecting the risk of developing pest resistance.
Paradigm
An acceptable refuge strategy should balance effectiveness and practicality (Caprio et al., 1999).
Implementing an impractical refuge has at least two consequences. The first is grower reluctance
or inability to plant a refuge that has unsatisfactory logistic or economic consequences to their
production system. A second consequence is segmenting growers so that only some farm sizes
can accommodate a refuge and the rest are unable to realize the economic and environmental
benefits of Bt maize.
Three main considerations have been taken into account:
a. The negligible risk of developing resistance posed by the actions of small farm
holders because of the very small total maize area that they represent.
b. The maize planting area below which implementing a generous 20% refuge becomes
an economic and logistic hurdle for growers.
c. To ensure that efforts in terms of education are used efficiently.
5 ha area threshold
The 5 ha area threshold was determined based on:
a. The agricultural landscape. Based on the four major maize-growing countries in the
EU (France, Germany, Italy, Spain), it can be inferred that:
-
Only a small proportion of the total maize area in the EU is cultivated on small
farms (less than 5 ha): in France, maize grown on small farms amounts to 8%, in
Germany 1%, in Italy 19% and in Spain 8% (Tables 1 and 2).
-
However, the “less than 5 ha” farms represent a significant proportion of the
maize farmers in the EU: in France 35%, in Germany 5%, in Italy 75% and in
Spain 47% (Tables 1 and 2).
This agricultural structure implies that the farming area, even specifically considering
maize, is very fragmented (for the general farming area fragmentation see Appendix 1).
b. The agricultural practices. It would not be economically sustainable for most of the
growers cultivating less than 5 ha of Bt maize to adopt a 20% refuge strategy
because:
-
Most maize seed is distributed in the EU using bags containing 50,000 to 80,000
kernels per bag, and seeding rates usually range from 70,000 to 80,000 kernels
per hectare
-
Maize producers usually spread the risk of an unpredictable growing season by
planting more than one seed product (hybrid name).
- Page 21/28 -
Planting a 20% refuge would in most cases force growers to buy excess seeds and
would be detrimental to their business.
c. The negligible risk of resistance. The proportion of less than 5 ha Bt maize area
represent a minority of maize land area with an equivalently small amount of
potential selective pressure because:
-
The fragmentation increases the probability that small maize fields will be
bordered by other crops, weedy/grass barriers or fallow land
-
The probability that target pests will routinely immigrate from multiple overwintering sites (previous maize) to maize planted during the following season
will be increased (Cordero et al., 1998; Hellmich et al., 1998; Losey et al., 2001).
Also, it can be expected that farms cultivating less than 5 ha of Bt maize will benefit from the
presence of non Bt maize on neighbouring farms which either did not adopt the Bt technology or
where refuges are implemented at the generous 20% level (the majority of the maize planting
area). Therefore, it is reasonable to consider that both the proportion and the fragmentation of the
less than 5 ha Bt maize area within the overall maize cultivated area in the EU makes it highly
unlikely for them to contribute to the development of pest resistance.
Since the risk of developing resistance is negligible, educational and monitoring efforts could be
better targeted to areas of Bt maize where there might be a potential for development of pest
resistance. Those efforts will much more efficiently be used to educate and survey the larger
growers that contribute to the great majority of maize production in the EU.
Conclusion
Effective and practical IRM can be achieved by adopting a generous 20% refuge on farms
planting Bt maize areas greater than 5 ha without compromising the potential for development of
pest resistance. This represents the largest and most relevant proportion of farmers for education
on insect resistance management, the largest fraction of the cultivated maize land area, and the
farms that are most likely to impact the potential development of pest resistance to Bt maize. In
addition, the 5 ha area threshold for farms provides a practical means of accommodating existing
agricultural and business practices without compromising the effectiveness of the refuge strategy.
- Page 22/28 -
Table 1: Distribution of maize cultivation over size classes of farm holdings for the four
largest maize-producing EU Member States.
Distribution of Maize Cultivation Over Farm Sizes
Farm size
Number of farms
Number of farms %
Area ha
Area %
< 5 ha
60,793
43%
132,700
8%
5 - 10 ha
29,224
20%
205,771
12%
France, cultivation for grain
10 - 20 ha
20 - 30 ha
27,099
11,363
19%
8%
377,469
274,463
22%
16%
30 - 50 ha
8,957
6%
338,852
19%
> 50 ha
5,265
4%
424,611
24%
Total ha
142,701
100%
1,753,866
100%
Number of farms
Number of farms %
Area ha
Area %
42,735
31%
105,078
8%
35,927
26%
257,719
19%
France, cultivation for silage
42,241
11,612
31%
8%
583,269
272,539
42%
20%
3,902
3%
138,109
10%
466
0%
28,214
2%
136,883
100%
1,384,929
100%
Farm size
Number of farms
Number of farms %
Area ha
Area %
< 2 ha
313
1%
144
0%
2 - 5 ha
2,700
7%
3,956
1%
5 - 10 ha
4,739
11%
11,571
3%
Germany, cultivation for grain
10 - 20 ha
20 - 30 ha
7,846
5,598
19%
13%
30,899
31,859
8%
9%
30 - 50 ha
8,851
21%
75,447
20%
50 - 100 ha
8,670
21%
121,141
33%
> 100 ha
2,974
7%
95,719
26%
Total ha
41,691
100%
370,735
100%
Number of farms
Number of farms %
Area ha
Area %
126
0%
74
0%
3,865
3%
4,007
0%
10,580
8%
15,003
1%
Germany, cultivation for silage
26,606
23,442
19%
17%
71,784
105,365
6%
9%
33,088
24%
239,159
20%
29,261
21%
334,796
28%
10,894
8%
432,655
36%
137,862
100%
1,202,844
100%
30 - 50 ha
10,150
3%
102,600
11%
50 - 100 ha
5,772
2%
127,220
11%
> 100 ha
2,837
1%
108,627
12%
Total ha
305,053
100%
900,328
100%
Farm size
Number of farms
Number of farms %
Area ha
Area %
< 2 ha
122,557
40%
84,489
9%
2 - 5 ha
80,549
26%
119,920
13%
5 - 10 ha
42,460
14%
127,947
14%
Italy, cultivation for grain
10 - 20 ha
20 - 30 ha
27,673
13,055
9%
4%
151,386
103,059
17%
11%
Number of farms
Number of farms %
Area ha
Area %
12,350
46%
9,500
3%
3,100
12%
15,300
5%
4,350
16%
40,700
12%
Italy, cultivation for silage
2,989
1,219
11%
5%
59,800
44,700
18%
14%
1,028
4%
41,100
13%
1,064
4%
56,200
17%
Spain, cutlivation for grain and silage
< 2 ha
2 - 5 ha
5 - 10 ha
10 - 20 ha
20 - 30 ha
30 - 50 ha
50 - 100 ha
Number of farms
30,915
35,912
24,242
23,158
8,657
7,393
6,910
Number of farms %
22%
25%
17%
16%
6%
5%
5%
Area ha
10,349
28,446
36,914
75,787
58,604
59,603
82,516
Area %
2%
6%
7%
15%
12%
12%
16%
Sources: France- Monsanto on ISTAT Source, Germany- Statistisches Bundesamt 1999, Italy- , Spain.
Italy – Monsanto on ISTAT Source
503
2%
60,300
18%
> 100 ha
5,586
4%
155,670
31%
26,602
100%
327,600
100%
Total ha
142,772
100%
507,889
100%
Table 2: Contribution of small farms to the extent of maize
grown in the EU (extracted from Table 1)
% of small farms* planting maize
(average)
% of maize planted on small farms*
(average)
France
35
8
Germany
5
1
Italy
75
19
Spain
47
8
* Farms of less than 5 ha
- Page 23/28 -
APPENDIX 3: Proposal for grower information material
INSECT RESISTANCE
MANAGEMENT
INFORMATION SHEET
Bt Maize
Bt maize has proven to be an important technology to help maize growers control
damaging insect pests and produce higher yields and better quality grain.
Insect Resistance Management (IRM)
To preserve the many benefits of Bt maize technology, the implementation of an
IRM plan is essential. An effective BT maize IRM plan includes the planting of a
non-Bt refuge (a block of non Bt maize) planted close to your Bt maize crop.
All Bt maize designed to control European corn borer and Mediterranean corn
borer require implementation of an IRM program according to the refuge size,
distance guidelines and insecticide usage described in this information sheet.
Refuge size requirements
If you plant greater than 5 hectares of Bt maize, total within or among fields you
must plant a refuge.
For each sowing, plant at least 2 hectares of non Bt maize for every 8 hectares of
Bt maize (minimum 20% non Bt refuge, maximum 80% Bt maize)
Refuge Distance Requirement
A non Bt maize refuge must be planted within 750 metres of each Bt maize field
- Page 24/28 -
Refuge Planting Options
As illustrated below, the appropriate size non Bt maize
refuge may be planted a number of ways:
Block Refuge
(adjacent)
A block of non Bt
maize adjacent to
the Bt maize field
Split Planter refuge
Strips of non Bt maize
at least 4 rows wide
within the Bt maize
field
Bt maize field
Block Refuge
(Within)
A block of non Bt
maize within the
Bt maize
Pivot Corners
Refuge
Non Bt maize in
pivot corners
within the Bt
maize field
Non Bt maize field
Wheat
- Page 25/28 -
Perimeter Refuge
Non Bt maize
surrounding Bt
maize field
Separate Field
Refuge
A separate field of
non Bt maize within
750 metres of the
Bt corn field
Insecticide Use in Bt and Non Bt refuge
Your Bt maize and non Bt maize refuge may be treated with conventional
insecticides ONLY if the target pest populations reach economic thresholds.
Microbial Bt insecticides must not be used within the refuge.
Refuge Management
In order to maximise the effectiveness of the refuge, you should manage your non
Bt maize and the Bt maize in a similar manner. This can be accomplished by planting
your non Bt maize as close to and at the same time as you Bt maize. In addition,
select non Bt hybrids and Bt hybrids that have similar growth and development
characteristics.
- Page 26/28 -
APPENDIX 4: Determining sample number and distribution for baseline susceptibility studies
and subsequent monitoring
The sample number and distribution of samples included in the baseline susceptibility study are
dependent on the factors that could increase the probability of insect adaptation to Bt maize. The
predictive factors used in this proposal are geographic and biological. Geographic variables
include: the total land area routinely planted to maize, the concentration of maize cultivation
within and among regions, and geological barriers to panmixia. Biological variables include: the
annual intensity of target pest infestations, the seasonal cycle of pests within a region, and our
best information on the biology and ecology of target pests. Integrating these factors does not
define the probability, but provides an indication of the relative likelihood of adaptation in
different regions, and this is a useful guide for the administration of resistance monitoring
resources.
Table 1 describes the distribution and concentration of maize production among the Member
States. In general, the probability of insect adaptation to Bt maize should be proportional to the
land area planted to Bt maize, the concentration of Bt maize in a particular region, and the
abundance of the target pest.
Table 1: Distribution and concentration of maize production among the Member States
Total land
area
(in 1,000ha)
54909
Arable land
area
(in 1,000ha)
18480
Maize land
area
(in 1,000ha)
3260
Germany
35697
11801
1517
12.9
DMK, 2001
Italy
30132
8192
1334
16.3
Spain
50488
12884
559
4.3
Austria
8386
1396
248
17.8
Consorzio Italiano per il
Telerilevamento
in
Agricoltura, 1999
Ministry of Agriculture,
1999
Agrarmarkt Austria, 1999
Netherlands
4153
977
246
25.2
Belgium
3052
852
221
26.0
Portugal
9191
2096
138
6.6
Greece
13196
1981
130
6.5
UK
24410
6625
101
1.5
National Statistical Institute
(CBS), 2001
(NIS), 2001
AMIS Seed, Kleffmann
(National Statistics & Seed
Industry Estimates), 2001
AMIS Seed, Kleffmann
(National Statistics & Seed
Industry Estimates), 2001
Eurostat, 2001
Denmark
4309
2364
79
3.3
Eurostat, 2001
Ireland
7029
1049
18
1.7
Teagasc, 2001
Luxembourg
257
60
11
18.3
Finland
33815
2143
-
-
(STATEC), 2000
Eurostat, 2001
Sweden
44996
2745
-
-
Eurostat, 2001
EU
Member
State
France
Total land area and arable land area sources: EuroStat 2001
- Page 27/28 -
Maize
Sources
concentration
for maize
(% of arable land)
land area
17.6
SCEES, 2001
Spain, France, Germany and Italy contain about 85% of all maize land area in the EU and in these
countries maize is grown on 4 to 18% of the total arable land. The remaining countries have
significantly smaller total areas routinely planted to maize and the concentration of maize on
arable land varies greatly among these countries.
The distribution of maize in dominant maize-producing countries is not evenly distributed, but is
aggregated in regions within countries. This aggregation suggests that resistance monitoring
efforts initially should focus on (but not necessarily be limited to) possibly four regions within the
EU where Bt Maize is likely to be cultivated. These regions will be defined as necessary to
maximize the efficiency of the monitoring programme.
- Page 28/28 -
MON 810 maize
May 2007
APPENDIX 3
Environmental monitoring plan from
Application for renewal of the
authorisation for continued marketing
of existing MON 810 maize products that
were authorized under Directive
90/220/EEC (Decision 98/294/EC) and
subsequently notified in accordance to
Article 20(1)(a) of Regulation (EC)
No 1829/2003 on genetically modified food
and feed
Part I
Technical Dossier
May 2007
Data protection.
This application contains scientific data and other information which are protected in accordance with Art. 31 of
Regulation (EC) No 1829/2003.
 2007 Monsanto Company. All Rights Reserved.
This document is protected under copyright law. This document is for use only by the regulatory authority to which this
has been submitted by Monsanto Company, and only in support of actions requested by Monsanto Company. Any other
use of this material, without prior written consent of Monsanto, is strictly prohibited. By submitting this document,
Monsanto does not grant any party or entity any right to license, or to use the information of intellectual property
described in this document.
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TABLE OF CONTENTS
Page
11. Environmental monitoring plan.................................................................. 4
11.1 General ..................................................................................... 4
11.2 Interplay between environmental risk assessment and
monitoring ................................................................................ 5
11.3 Case-specific GM plant monitoring ......................................... 6
11.4 General surveillance of the impact of the GM plant .............. 6
11.5. Reporting the results of monitoring........................................ 23
References........................................................................................................ 24
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LIST OF FIGURES
Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
General surveillance responsibilities.................................................10
Structure of the monitoring data .......................................................12
Structure of the data in the GS database ..........................................13
Data matching from different sources ...............................................14
Statistical model for analysing monitoring characters .....................22
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11. Environmental monitoring plan
11.1 General
As the scope of this application under Regulation (EC) No 1829/2003
includes the use of MON 810 for the cultivation of varieties in the
European Union (E.U.), a monitoring plan conforming with
Annex VII of Directive 2001/18/EC is included, as required by
Articles 5(5) and 17(5) of the said Regulation.
According to Annex VII of Directive 2001/18/EC, the objective of a
monitoring plan is:
- To confirm that any assumption regarding the occurrence and
impact of potential adverse effects of the GMO or its use in the
environmental risk assessment (e.r.a.) are correct,
and
- To identify 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.
This plan describes the strategy and methodology to monitor the
placing on the market of MON 810.
- It is based on the characteristics of MON 810, the scale of its use
and the range of relevant environmental conditions of the areas
where MON 810 is released
-
It facilitates the observation, in a systematic manner, of the
release of MON 810 in the receiving environment and the
interpretation of these observations with respect to safety on
human and animal health and on the environment
-
It identifies the type of effects and variables to be monitored and
the time-period for measurements
-
It describes the tools for data sampling, analysing and reporting,
whereby it focuses on farm questionnaires as the main
surveillance tool
-
It identifies who will carry out the various tasks outlined in the
monitoring plan and who is responsible for ensuring that the
monitoring plan is set into place and carried out appropriately. It
also ensures that there is a route by which the party placing the
genetically modified (GM) plant on the market, the European
Commission and the Member States will be informed on any
observed adverse effects on human health and the environment
-
It enables the party placing the GM plant on the market to
inform the Member State and to take the measures necessary to
protect human and animal health and the environment, where
appropriate.
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According to the guidance notes supplementing Annex VII to
Directive 2001/18/EC, the monitoring strategy aims to identify the
potential adverse effects that may arise from placing MON 810 on
the market, the degree to which these effects need to be monitored,
and gives an approach and a time scale over which to monitor.
11.2 Interplay between environmental risk assessment and
monitoring
An e.r.a. of MON 810 (see Part I, Section D.9 of this application) was
undertaken in the context of the scope of the application, that is, for
consent for use of MON 810 in the E.U. as any other maize,
including the cultivation of MON 810 varieties.
Analysis of the characteristics of MON 810 and comparison to the
experience with cultivation of conventional maize within the E.U.
has shown that the risk for potential adverse effects on human and
animal health and the receiving environment, resulting from the use
of MON 810 in the E.U., including the cultivation of MON 810
varieties and use thereof as any other maize, is consistently
negligible relative to:
- Persistence and invasiveness
- Selective advantage or disadvantage
- Potential for gene transfer
- Interactions between the GM plant and target organisms
- Interactions of the GM plant with non-target organisms
- Effects on human health
- Effects on animal health
- Effects on biogeochemical processes
- Impacts of the specific cultivation, management and
harvesting techniques
Therefore, the overall environmental risk posed by this GM plant is
negligible, and no specific strategies for risk management are
required.
Since the conclusions of this e.r.a. are derived from the results of
scientific studies rather than major assumptions, it is proposed that
no case-specific post-marketing monitoring (CSM) actions, typically
aimed at testing assumptions made in this assessment, would be
warranted or required.
Instead, the monitoring concentrates on general surveillance (GS) to
allow the identification of adverse effects of MON 810 or its use on
human health or the environment, which were not anticipated in the
e.r.a.
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11.3 Case-specific GM plant monitoring
The objective of case-specific monitoring is to confirm that any
assumptions regarding the occurrence and impact of potential
adverse effects of the GMO or its use that have been identified in the
e.r.a., are correct.
Since the conclusions of the e.r.a. (included in Part I, Section D.9 of
this application) consistently show that the placing on the market of
MON 810 poses negligible risk to human and animal health and the
environment and since the conclusions of this e.r.a. are derived from
the results of scientific studies rather than major assumptions, it is
proposed that no case-specific post-marketing monitoring actions,
typically aimed at testing assumptions made in this assessment,
would be warranted or required.
MON 810 is, however, commercialised alongside company
stewardship programmes such as the IRM plan presented in
Appendix 1 (see also Section D.9.9.), and the plan for general
surveillance, described in Section D.11.4.
11.4 General surveillance of the impact of the GM plant
11.4.1 Approach
The objective of general surveillance is to identify 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.
General surveillance is largely based on routine observation
and implies the collection, scientific evaluation and reporting
of reliable scientific evidence, in order to be able to identify
whether unanticipated, direct or indirect, immediate or
delayed adverse effects might have been caused by the placing
on the market of a GM plant in its receiving agricultural or
non-agricultural environment. By nature, the prediction of
unanticipated effects does not lend itself to the formulation of
clear scientific hypotheses, and therefore it will need adapted
scientific methodology, as described in Section 4.3.
The GS plan detailed below is in line with the Biotech
Industry approach, described by (Tinland et al., 2006).
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11.4.2 Strategy
The objective of Directive 2001/18/EC is to protect “human
health and the environment”. Although it is difficult to define
monitoring parameters, concrete and informative monitoring
characters for general surveillance can be derived from more
specific protection goals and their areas of potential impact
covering the term “human health and the environment”
(Wilhelm et al., 2003).
Protection goals are:
- Ecological systems and biodiversity
- Soil function
- Sustainable agriculture
- Plant health
- Human and animal health
Areas of potential impact on these protection goals to be
monitored might be, respectively:
- Invasiveness
- Soil fertility, soil biology, mineralization, loss of soil
(erosion, compression)
- Use of fertilisers, use of pesticides, persistence,
cultivation methods
- Plant diseases, pests, weeds
- Toxicity, pathogenicity, allergenic potential, nutrition
quality
From these areas of potential impact, a range of monitoring
characters have been derived for general surveillance of
MON 810 (for details, see Section 11.4.3.2).
Possible effects in these areas might also be caused by other
influencing factors, especially:
- Climatic conditions
- Agricultural practice (choice of crop, planting time,
cultivation methods, use of herbicides, insecticides,
fungicides, fertilizer, rotational practices, etc.)
- Incidence of plant pests, plant diseases and weeds
- Landscaping and restructuring
- Other human activity leading to e.g. pollution
Therefore, a range of influencing factors to be monitored
additionally has been identified (see Section 11.4.3.2).
General surveillance focuses on the geographical regions
within the E.U. where MON 810 is grown, and takes place in
representative environments, reflecting the range and
distribution of farming practices and environments exposed to
MON 810 plants and their cultivation.
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To collect relevant data on monitoring characters and
influencing factors, a cyclic monitoring process using different
sources of data is established, involving:
- Individuals or organizations normally involved in
agriculture, or whose activities are connected to
agriculture, the environment and human and livestock
health
- Company stewardship programmes
- Selected, existing networks
- Data from other sources: internet, scientific
publications, …
Where there is scientifically valid evidence of a potential
adverse effect (whether direct or indirect) linked to the
genetic modification, further evaluation of the consequence of
that effect should be science-based and compared with
baseline information. Relevant baseline information reflects
prevalent agricultural practice and the associated impact of
these practices on the environment.
In many cases it may not be possible to establish a causal link
between a potential adverse effect and use of a particular GM
plant.
11.4.2.1 Definition of baselines and effects
The identification of potential adverse effects is based
on historical knowledge and experience of users of
MON 810 in relation to their use of conventional
maize, prior to the introduction of MON 810 and the
simultaneous cultivation of conventional maize on
the same or neighbouring farms.
An effect is defined as an alteration that results in
values that are outside the baseline variation given
the continuous dynamics of European agriculture,
agricultural practices, rural environment and
associated biota.
A major challenge of general surveillance is
determining whether:
- an unusual effect has been observed
- the effect is adverse and
- the adverse effect is associated with the GM
plant or its cultivation.
11.4.2.2 Time-period
The time-period for general surveillance should be in
line with the period of consent, i.e. maximum 10
years.
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11.4.2.3 Responsibilities
The party placing the GM plant on the market is
responsible for informing the European Commission
and the Member States of any adverse effects
observed during general surveillance.
The party placing the GM plant on the market will
primarily consider general surveillance in the areas
where that GM plant is grown and monitor for any
adverse effects of its cultivation at farm level (Figure
1). When appropriate, the data collected from
different regions can be analysed for regional
patterns or trends.
However, surveillance for adverse impacts of GM
plants at complex regional and/or national levels is
beyond the scope of farm monitoring or the direct
capability of the party placing the GM plant on the
market. Therefore, the general surveillance at this
level is considered to be a national/European
responsibility, as illustrated in Figure 1.
Increasing complexity and interaction of GM plants
with other land management systems may be studied
in other ways. Utilising existing surveillance systems
established
by
land-use
and
environmental
organisations was identified by EFSA as a potential
approach to complement the general surveillance.
This approach may allow for a comprehensive view of
GM
cultivation
in
a
broader
agricultural
environment.
To conclude, the party placing the GM plant on the
market makes use of farm questionnaires for GMOfocused GS at farm level. The national Competent
Authorities may integrate the results from regional
agricultural and environmental surveys reported to
them in their evaluation of potential adverse effects
to identified protection goals in the broad
environment. GMO-effects are assessed as one of the
many potential influencing factors.
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Potential adverse
impact at National/
European level
Countries/ EU
Public
sector
Potential adverse
impact at
regional level
Landscape
Farming system
Potential adverse
impact at
farm level
Potential adverse crop
and management impact
Figure 1.
Field
Party placing
the GM crop
on the market
General surveillance responsibilities
11.4.3. Monitoring methodology
11.4.3.1 Framework - Data structure and management
11.4.3.1.1 Definitions
For the purpose of general surveillance, the following
definitions are used (Figure 2).
o Test factor = the cultivation of the GM plant
versus baseline agricultural practices for cultivation
of the conventional counterpart plant
The objective of general surveillance is to investigate
whether the cultivation of GM plants has an adverse
impact on operational health and the environment.
The identification of a potential adverse effect will be
based on the baseline, as defined in Section 11.4.2.1.
o Monitoring characters:
Given the definition of the test factor, the monitoring
activities focus on parameters which might be
influenced by cultivation of the GM plant and might
show a measurable effect when compared to the
impact of baseline practices on that parameter. These
parameters are good candidate monitoring characters
if they can be associated with desirable protection
goals (see Section 11.4.2) and if they can be defined in
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such a way that they could indicate potential adverse
effects or deviations from accepted and baseline
ranges.
o Monitoring object = Field grown with the GM
plant and its immediate surroundings
The party placing the GM plant on the market will
primarily consider general surveillance in the places
where the GM plant is grown and monitor for any
adverse effects of its cultivation at farm level. Time
and space coordinates (e.g.: country, postal code,
year, …) define the monitoring object. The objects to
be monitored have to be the cultivation areas (fields),
since measurable effects first are to be expected here
and because fields present the smallest unit where
monitoring characters can be observed in relation to
the test factor and where the influencing factors (as
defined below), especially cultivation practices, can
be assumed to be equal.
o Influencing factors:
As agro-ecosystems are influenced by numerous
factors, the selected monitoring characters may be a
function of many factors besides the test factor.
These other factors are called influencing factors or
covariates and can be subdivided in three categories:
o Adjustable factors: e.g. planting time,
agronomic practices, amount of pesticides
and fertilizer used, …
o Random factors: e.g. disease, pest and weed
pressure, weather, …
o Fixed factors:
rotation, …
e.g.
soil
type,
previous
Ideally, all influencing factors that may have an
impact on the character of observation must be
collected, as this information may reveal the
independent or confounding cause of observed
variation in a monitoring character.
Each observed parameter has to be assigned to one of
the possible types of parameters: i.e. as intrinsic
monitoring character (e.g. development, disease,
weed presence) or influencing factor (e.g. agricultural
practices, soil parameters,…). The intrinsic
monitoring characters are those of interest as they
may elicit potentially adverse effects in terms of
protection goals, whereas the values of the
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influencing factors are recorded to analyse the
monitoring characters for potential variation, caused
by other factors than the test factor.
The cultivation of the GM plant as opposed to the
conventional counterpart plant may only be one
possible reason or contributor for a potential adverse
effect.
Therefore,
all
background
data
on
environment and soil characteristics of the site and
the established cultivation practices have to be
identified and evaluated (e.g. whether observed
variations are due to weather conditions, seeding
date etc. rather than to the GM plant). Statistical
analysis has the task to identify the reasons for
effects within the monitoring characters and to
separate GM plant effects from influences due to
other environmental and cultivation factors and from
random variation.
geographic coordinates
Test Factor
Monitoring object
= Field and immediate Surroundings
Monitoring characters
Adjustable Factors
= GMO vs. baseline
= Parameters
Random Factors
Fixed Factors
Influencing Factors
Figure 2.
Structure of the monitoring data
In conclusion, general surveillance of a GM plant
aims to record and analyze key characteristics of GM
production fields and their immediate environment
that may reveal unanticipated adverse effects of the
cultivation of the GMO on defined environmental
protection goals through a statistical analysis of
carefully selected monitoring characters, and other
influencing factors that may confound the observed
12
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variation of the monitoring factors but which are not
related to the GM plant.
11.4.3.1.2 General surveillance database
To structure and store the general surveillance data,
a GS database is used. Within this database it is
possible to categorise the data as influencing factor,
monitoring character, etc., to pool and match
surveillance data (e.g. for a connection in time and
space) and to characterise them for statistical
analysis (Figure 3).
Codes define the
monitoring object
Code/ ID
Time
Levels/ Measures of the Influencing
Factors
Location
Adjustable factors
Fixed factors
Tillage
Insecticides
Soil
type
Soil
quality
Random
factors
Disease
pressure
Monitoring
character
Occurence of plant
disease
2007-02-MONES-AB-01-01
2007
Spain,
Albacete
yes
yes
F3
xy21
high
more
2007-02-MONFR-VF-04-01
2007
France,
Galiax
yes
no
F4
xy22
low
as usual
2008-02-MONPT-AB-02-01
2008
Portugal,
Odemira
no
yes
F7
xy11
as usual
as usual
Definition of time
and space
coordinates
Figure 3.
Measures of the
Monitoring Object
Structure of the data in the GS database
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Party placing the GM crop on the market
questionnaires
statistics
report
GS database
e.g. plant development
Competent
Authorities
report
e.g. rain [mm]
database
Existing Networks
report
e.g. plant diseases
database
Public Sector
key connection
(time and location)
Figure 4.
Data matching from different sources
A GS database is developed to manage the data from
farm questionnaires. It will also be connectable to
data from other sources (Figure 4). If a potential
adverse effect is identified, the party placing the GM
plant on the market can gather additional data to
understand whether this adverse effect is associated
with the GM plant. Therefore, the party placing the
GM plant on the market can use other sources of
information,
discussed
in
more
detail
in
Section 11.4.3.2.2. The key for data pooling from
different sources should be their temporal and spatial
coordinates. Therefore, all data sets have to be
identified by their origin – the date and location of
survey. A matching of spatiotemporal coordinates of
datasets coming from different sources allows, for
example, assessing whether an observed negative
effect on plant development reported in a subset of
questionnaires for a certain region, can be related to
a higher occurrence of plant diseases in that region
observed by a public plant protection network.
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11.4.3.2 Identification of monitoring parameters and
description of tools
For general surveillance of MON 810, the party
placing MON 810 on the market uses several tools.
The central tool is an annual farm questionnaire
addressed to a subset of farmers cultivating
MON 810. Additionally, information from other
sources (company stewardship programmes, scientific
literature, official websites and existing observation
networks) will be incorporated, where appropriate.
11.4.3.2.1 Farm questionnaires
Farmers are the closest observers of the cultivation of
the GM plants and they already collect information
on the cultivation and management of their crops at
farm level. Therefore, they can give details on GM
plant-based
parameters
(referring
to
species/ecosystem biodiversity, soil functionality,
sustainable agriculture, or plant health) and on
background and baseline environmental data (e.g.
soil parameters, climatic conditions, general crop
management data such as fertilisers, crop protection,
crop rotations and previous crop history).
Additionally,
farmers
may
give
empirical
assessments which can be useful within general
surveillance to reveal unanticipated deviations from
baseline variation for the crop and cultivation area in
question, based on their historical knowledge and
experience and parallel non-GM cultivation
(Schiemann et al., 2006).
A questionnaire addressed to the GMO cultivating
farmers is a monitoring tool that is specifically
focused at farm level. EFSA explicitly considers
questionnaires a useful method to collect first hand
data on the performance and impact of a GM plant
and to compare the GM plant with conventional
plants (EFSA, 2006a).
Since the party placing MON 810 on the market uses
farm questionnaires as the key tool for monitoring of
MON 810, a questionnaire has been designed to ask
farmers for their observations and assessment in and
around the GM cultivated field in comparison to a
baseline, being their own historical knowledge and
experience. The unit of observation is the field or
group of fields a certain farmer is cultivating
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MON 810 on. The questionnaire focuses on four
sections:
1. Maize grown area
This section is to obtain information on the location
and the size of the MON 810 cultivation area to
identify the fields and its geographic coordinates.
Furthermore, the farmers are asked to specify the
soil characteristics and the season’s general pest
pressure to obtain background and baseline data
(independent from GM or non-GM cultivation).
2. Typical agronomic practices to grow maize on the
farm
This section refers to the cultivation practices on
the farm to identify the levels of adjustable factors
being applied. This paragraph is to refer to the non
GM area. The goal is to find out what the normal
practices in conventional cultivation are to see
later whether they differ from those in GM areas.
3. Observations on MON 810 (YieldGard®)
This section poses farmers questions regarding
their agricultural practices in MON 810 fields, on
the characteristics of MON 810 in the field, on the
disease, pest and weed pressure and the occurrence
of wildlife in MON 810. Additionally, they are
asked about the performance of the animals fed
MON 810. Farmers can specify their observations
and assessments compared to conventional maize
by giving categories like “as usual”, “more” or “less”
and give specifications for answers differing from
“as usual”. Furthermore additional remarks or
observations may be taken down.
4. Implementation of GM plant specific measures
This section is
stewardship data.
to
obtain
compliance
and
In conclusion, the questionnaire is designed to collect
background and baseline data as well as GM-based
monitoring
characters.
An
example
of
a
questionnaire for MON 810 is presented in
Appendix 2.
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11.4.3.2.2 Existing observation networks
Many of the existing monitoring systems and
networks collecting environmental data are unlikely
to always provide relevant data for monitoring the
impact of GM plants. The design of the existing
monitoring programs, the targets (e.g. birds, plant
protection, etc.), the time, frequency and scale of data
collection, sampling, analysis and reporting methods
may not suit the monitoring of GM plants because
they have been designed for other purposes.
Moreover, the existing monitoring systems will differ
from country to country and it may not be feasible to
modify existing monitoring systems in order to make
them suitable for general surveillance focused on GM
plants.
Nevertheless, the use of existing networks to provide
surveillance information is seen as a key aspect for
ensuring that sufficient observers are available to
identify and report possible unanticipated adverse
effects, as well as ensuring methodological
consistency and optimising the expenditure of
resources. This would include existing observation
programs in the fields of agriculture, the nonagricultural environment, occupational health and
livestock welfare. As stated in the EFSA opinion on
post-market environmental monitoring (EFSA,
2006b), the use of national environmental monitoring
programmes is outside of the management and
control by an individual applicant and thus it cannot
be the task of an applicant alone to use, modify or
improve existing surveillance systems. The party
placing the GM plant on the market therefore
proposes to broaden the reporting obligations of the
operators of these programmes to the Competent
Authorities for GMO cultivation and risk
management, who can then consider any
scientifically founded information from these regional
agricultural and environmental surveys collected in
the countries where MON 810 is grown together with
the GS report of the party placing MON 810 on the
market. Taken together, the information reported to
them will allow the Competent Authority to evaluate
potential adverse effects to identified protection goals
in the broad environment. GMO-effects are assessed
as one of the many potential influencing factors.
Some examples of national environmental monitoring
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programmes that could be connected to the potential
needs of general surveillance come from Switzerland
(Bühler, 2006) and France (Delos et al., 2006).
It has to be noted that established routine
surveillance networks (providing monitoring of
agriculture, variety registration monitoring, plant
protection, plant health and soil surveys and
ecological/environmental monitoring) might provide
useful data on background or baseline characters on
a landscape or national level (e.g. climatic conditions,
cultivation practices) or on monitoring characters
(plant diseases or pests, invasiveness, weeds). The
party placing the GM plant on the market may
therefore consider to use information from this type
of networks on an ad hoc basis (e.g. if a potential
adverse effect is reported in a subset of
questionnaires in a certain region) to assess whether
this effect is associated with the GM plant or with
another influencing factor. Networks for the
agricultural and the non-agricultural environment as
well as for human and livestock health might be
consulted, when suitable. The main criteria for
selecting the appropriate networks have to be: data
quality, quantity, compatibility and accessibility (to
the party placing the GM plant on the market). In
some European countries (e.g. Germany, France),
there are scientific projects in place analysing the
existing networks for their suitability for GMO
monitoring (Mönkemeyer et al., 2006).
In addition, in the E.U., networks of international
traders and grain processors, will be asked to provide
annual feedback to the authorisation holder, through
EuropaBio1, on potential adverse effects associated
with the import and handling of MON 810.
11.4.3.2.3 Company stewardship programs
A continuous supply and distribution network
extends from the technology provider, via
intermediate distribution, to the end-user. Through
their sales and technical organisations, key
participants, especially those companies involved in
farm sales, would be regular visitors to fields where
GM plants would be cultivated. Experience has
shown that this network ensures a continuous and
efficient communication link from the grower to the
1
http://www.europabio.org/InfoOperators/General%20Surveillance.htm
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technology provider, especially in relation to
complaints about product performance, and thus
would provide a key surveillance network for possible
adverse effects.
The stewardship commitment of the authorisation
holder is detailed in the Technoloqy Use Guide
(2007)2.
11.4.3.2.4 Other sources
In addition to the above-mentioned general
surveillance actions directed to MON 810 growers,
international traders, grain processors, users of
maize grain, and other stakeholders, the party
placing MON 810 on the market will actively monitor
existing information sources such as official websites,
scientific publications and expert reports on GMOs in
order to identify, collate and follow-up on potentially
adverse observations made for this maize or any
other relevant information, in particular with respect
to occupational health, animal feed safety or putative
ecological effects of the release of this maize.
11.4.3.3 Areas and size of sampling
11.4.3.3.1 Areas to be monitored
Following the approval of MON 810 for cultivation in
the E.U., a portion of the maize cultivated area has
been replaced by MON 810. Estimates of the total
annual maize production and typical grain
production tonnages per Member State are given in
Part I, Section D.7.7 of this renewal application.
Significant areas of maize production in Europe
include the Danube basin from southwest Germany
to the Black Sea and southern France through the Po
Valley of northern Italy. The glyphosate tolerance
trait has utility in a wide range of agricultural
environments. Therefore, the introduction of
MON 810 is not confined to specific geographical
zones. The introduced agronomic trait does not alter
patterns or volumes of maize production and
consumption.
Monitoring activities are mainly focused on areas
where MON 810 has a high market penetration.
2
http://www.monsanto.com/monsanto/us_ag/layout/stewardship/tug/default.asp
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11.4.3.3.2 Number of farm questionnaires sampled
Monitoring characters are surveyed to assess
whether GM cultivation results in any adverse effects
compared to conventional maize. Therefore, mainly
ordinally distributed characters with three possible
values will be recorded within the questionnaires.
The ordinal monitoring characters (“as usual”/
“more”/ “less”) have one level for the adverse
situation (mostly: “less”), one level for positive effects
(mostly “more”) and one level when no changes
occurred (“as usual”). For the purpose of monitoring,
only the adverse effects are of interest, so that the
data will be transformed into binary distributed
characters (“adverse effect”/ “no or positive effect”).
The statistical procedure to evaluate these binary
response frequencies is a statistical test with the null
hypothesis (H0) being that the response frequencies
for “adverse effects” do not exceed the threshold limit
of 5% (no statistically significant adverse effects of
GM cultivation) and the alternative hypothesis (H1)
being that there are more than 5% of “adverse effect”
answers.
The experimental design for the tests with the given
hypotheses will be done for determined values for the
errors of the first and second kind. In each statistical
test, decisions for or against the null hypothesis are
taken with α and β error probabilities, where α fixes
the probability of rejecting H0 although it is true and
β gives the probability of accepting H0 although it is
wrong. In monitoring it is important to detect
adverse deviations from the usual situation, i.e. to
accept H0 with high probability (small α probability).
On the other hand, a wrong rejection of H0 leads to
far-reaching consequences and therefore, should also
be allowed with small probability β, i.e. the power of
the test should be as high as possible. Therefore, the
sample size was planned with α = 0.01 and β = 0.01.
The tolerance limit for the response frequency
following the results from the German pilot study
(Schmidt et al., 2004; Wilhelm et al., 2004) was set to
0,925, which results into a necessary sample size of
2.008 questionnaires. Under consideration of a drop
out quota of about 10 – 20% due to bad data quality
or missing questionnaires Monsanto plans to survey
2.500 questionnaires spread over the monitoring
period of 10 years.
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11.4.3.4 Frequency of sampling
Farm questionnaires are distributed, completed and
collated annually.
Due to little cultivation in the beginning of the
authorisation period the survey has started with 132
planned 2500 questionnaires in 2005. This number
as reached the 250 in 2006, which is the forthcoming
annual target number.
11.4.4 Analysis
Using statistical procedures, the monitored characters can be
analysed for differences or significant adverse deviations from
the usual situation and the causes for these deviations can be
detected in the whole complex system. With this approach,
the test factor “GMO vs baseline” and any effects (differences)
in the monitoring characters can be analysed for their relation
to this cultivation.
21
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Code/ ID
Time
Location
Adjustable factors
Fixed factors
Tillage
Insecticides
Soil
type
Soil
quality
Random
factors
Disease
pressure
Monitoring
character
Occurence of plant
disease
2007-02-MONES-AB-01-01
2007
Spain,
Albacete
yes
yes
F3
xy21
high
more
2007-02-MONFR-VF-04-01
2007
France,
Galiax
yes
no
F4
xy22
low
as usual
2008-02-MONPT-AB-02-01
2008
Portugal,
Odemira
no
yes
F7
xy11
as usual
as usual
thresholds?
descriptive analysis
biological variability
significant differences?
Causal connection?
Figure 5.
Statistical model for analysing monitoring characters
The whole analysis procedure takes place in three steps
(Figure 5):
1. Analysis of the monitoring characters for any
significant deviations from normal biological variation
reported by the farmer, possibly referring to an adverse
impact on the protection goal
2. Analysis of influencing (adjustable, random or fixed)
factors for any significant deviations from conventional
crop cultivation
3. Analysis of the relationships between monitoring
characters and influencing factors for any possible
causes of effects by the influencing factors
The statistical test procedures to evaluate the data for a
possible adverse effect work on the basis of baseline value
centred tolerance intervals for the monitoring characters and
with probabilities for erroneous decisions. If they confirm any
unusual effect with high significance probability, the cause for
this effect has to be identified within the test and the
influencing factors. Generally, the possible significant
influence of any of the factors being considered in data
collection can be investigated by several statistical methods
(factor analysis, regression analysis, analysis of variance).
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This is done to exclude any bias in the data and to ascertain
any influences and correlations for confirming real test factor
effects (Schmidt et al., 2006).
It is known that the environmental conditions have an
influence on the monitoring characters. Monitoring will
therefore be done over a significant period of time and at a
representative number of sites, and the factors “time” and
“space” will be analysed especially for their influence on the
monitoring characters. Splitting the data in time series or
spatially looking for shifts could also help in assessing
whether or not the potential adverse effect is associated to the
GM plant.
11.5. Reporting the results of monitoring
Any recorded observations of adverse findings that are linked to the
cultivation and/or use of this maize, which come to the attention of
the party placing the GM plant on the market, will receive careful
analysis in real time and re-mediating action, where applicable.
Adverse reports will be discussed in the mandatory general
surveillance report. The general surveillance reports will be sent to
the European Commission, which will distribute to all Competent
Authorities in the E.U. General Surveillance reports will be
prepared on an annual basis, except in case of adverse findings that
need immediate risk mitigation, which will be reported as soon as
possible.
Since monitoring of GM plants is a new topic and a creative process,
the monitoring plan and especially the questionnaires can be
improved based on experience from year to year.
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References
Bühler, C. (2006) Biodiversity monitoring in Switzerland: what can we learn for
general surveillance of GM crops? J. Verbr. Lebensm., 1, 37-41.
Delos, M., Hervieu, F., Folcher, L., Micoud, A. and Eychenne, N. (2006) Biological
surveillance programme for the monitoring of crop pests and indicators in
France. J. Verbr. Lebensm., 1, 30-36.
EFSA. (2006a) Guidance document of the scientific panel on genetically modified
organisms for the risk assessment of genetically modified plants and
derived food and feed. The EFSA Journal, 99, 1-100.
EFSA. (2006b) Opinion of the Scientific Panel on genetically modified organisms
on the post-market environmental monitoring (PMEM) of genetically
modified plants. Question No EFSA-Q-2004-061, 319, 1-27.
Schiemann, J., Wilhelm, R., Beissner, L., Schmidtke, J. and Schmidt, K. (2006)
Data acquisition by farm questionnaires and linkage to other sources of
data. J. Verbr. Lebensm., 1, 26-29.
Schmidt, K., Schmidtke, J., Wilhelm, R., Beissner, L. and Schiermann, J. (2004)
Biometrische Auswertung des Fragebogens zum Monitoring des Anbaus
gentechnisch veränderter Maissorten - Statistische Beurteilung von
Fragestellungen
des
GVO-Monitoring.
Nachrichtenbl.
Deut.
Pflanzenschutzd., 56, 206-212.
Tinland, B., Janssens, J., Lecoq, E., Legris, G., Matzk, A., Pleysier, A., Wandelt,
C. and Willekens, H. (2006) Implementation of general surveillance in
Europe: the industry perspective. J. Verbr. Lebensm., 1, 42-44.
Wilhelm, R., Beissner, L. and Schiermann, J. (2003) Concept for the realisation of
a GMO monitoring in Germany. Federal Biological Research Centre for
Agriculture and Forestry, Institute for Plant Virology, Microbiology and
Biosafety.
Wilhelm, R., Beissner, L., Schmidt, K., Schmidtke, J. and Schiemann, J. (2004)
Monitoring des Anbaus gentechnisch veränderter Pflanzen - Fragebögen
zur
Datenerhebung
bei
Landwirten.
Nachrichtenbl.
Deut.
Pflanzenschutzd., 56, 184-188.
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MON 810 maize
May 2007
APPENDIX 4
Year
Event
01Partner
Interviewer Farmer Area
Country
General Surveillance of YIELDGARD CornBorer (MON 810)
1. Maize grown area
1.1. Location:
Country ..................
County ...................
1.2. Size of maize area and varieties grown:
Total area of all maize cultivated on farm (ha)…..........
Total area of MON 810 cultivated on farm (ha)….........
List up to five MON 810 varieties planted this season:
1.
2.
3.
4.
5.
List up to five conventional varieties planted this season:
1.
2.
3.
4.
5.
1.3. Soil characteristics of the maize grown area:
Mark the predominant soil type of the maize grown area (soil texture):
O
O
O
O
O
O
Very fine (clay)
Fine (clay, sandy clay, silty clay)
Medium (sandy clay loam, clay loam, sandy silt
Medium-fine (silty clay loam, silt loam)loam)
Coarse (sand, loamy sand, sandy loam)
No predominant soil type (too variable across the maize grown area on
the farm)
O I do not know
Characterise soil quality of the maize grown area (fertility):
O Below average - poor
O Average - normal
O Above average -good
Year
Event
01Partner
Country
Organic carbon content (%)..........
1.4. Local pest and disease pressure in maize:
Characterise this season’s general pest pressure on the maize cultivated area:
Diseases (fungal, viral)………………. O Low
O As usual
O High
Pests (insects, mites, nematodes)…. O Low
O As usual
O High
Weeds…………………………………. O Low
O As usual
O High
2. Typical agronomic practices to grow maize on your farm
2.1. Irrigation of maize grown area:
O Yes
O No
2.2. Major rotation of the maize grown area:
previous year:
two years ago:
2.3. Soil tillage practices:
O No
O Yes (mark the time of tillage:
2.4. Maize planting technique :
O winter
O spring)
O Conventional planting
O Mulch
O Direct sowing
2.5. Mark all typical weed and pest control practices in maize at your
farm:
O Herbicide(s)
O Insecticide(s)
O Fungicide(s)
O Mechanical weed control O Use of biocontrol treatments (e.g. Trichogramma)
O Other, please specify:…..
2.6. Application of fertiliser to maize grown area:
O Yes
O No
2.7. Typical time of maize sowing range (DD:MM – DD:MM):
/
-
/
Year
Event
01Partner
Country
2.8. Typical time of maize harvest range (DD:MM – DD:MM):
Grain maize:
/
Forage maize:
/
-
/
/
3. Observations of YieldGard® CornBorer (MON 810)
3.1. Agricultural practices in MON 810 (compared to conventional maize)
Did you change your agricultural practices in MON 810 compared to conventional
maize? If any of the answers is different from «As usual», please specify the change.
Did you plant MON 810 earlier or later than conventional maize?
O As usual
O Earlier
O Later
Because:
Did you change your soil tillage or maize planting techniques to plant MON 810?
O As usual
O Changed, because:
Full commercial name of insecticides you applied in MON 810 field
1.
2.
3.
4.
Full commercial name of herbicides you applied in MON 810 field
1.
2.
3.
4.
Full commercial name of fungicides you applied in MON 810 field
1.
2.
3.
4.
Year
Event
01Partner
Country
In 2006, how were the weed and pest control practices in MON 810 when compared
to conventional maize?
Insecticide: O Similar
O Different, because:
Herbicide:
O Similar
Fungicide:
O Similar
In 2006, how were the fertiliser application practices in MON 810 when compared to
conventional maize?
O Similar ..........O Changed, because:
In 2006, how were the irrigation practices in MON 810 when compared to
conventional maize?
O As usual ..........O Changed, because:
Did you harvest MON 810 earlier or later than conventional maize?
O As usual .....…O Earlier.…..O Later
Because:
3.2. Characteristics of MON 810 in the field (compared to conventional maize)
Germination vigour…..… O As usual
O More vigourous
O Less vigourous
Time to emergence…….. O As usual
O Accelerated
O Delayed
Time to male flowering… O As usual
O Accelerated
O Delayed
Plant growth and
development……………. O As usual
O Accelerated
O Delayed
Incidence of stalk/root
lodging…….…………….. O As usual
O More often
O Less often
Time to maturity………… O As usual
O Accelerated
O Delayed
Yield……………………… O As usual
O Higher yield
O Lower yield
Occurrence of MON810
Volunteers from previous O As usual
year planting (if relevant)
O More often
O Less often
Year
Event
01Partner
Country
If any of the answers above is different from « As usual », please specify:
Please detail any additional unusual observations regarding the MON 810
maize during its growth:
3.3. Characterise the disease pressure in MON 810 fields (compared to
conventional maize)
Overall assessment of disease susceptibility of MON 810 compared to conventional
maize (fungal, viral diseases):
O Normal
O More disease
O Less disease
If the above answer is different from «Normal», please specify the difference in
disease susceptibility in the list and the commentary section below:
1. Fusariosis (Fusarium spp)………………………………….
2. xxx ……………………………
3. xxx ..
4. xxx …………………………………….
5. Anthracnose (Colletotrichum graminicola)………………..
6. Other:
……..
O
O
O
O
O
O
More
More
More
More
More
More
O
O
O
O
O
O
Less
Less
Less
Less
Less
Less
Specific to each country
Additional comments :
3.4. Characterise the INSECT pest control in MON 810 fields (compared to
conventional maize)
On the two insects controlled by MON 810, overall efficacy of the GM varieties on:
1. European corn borer (Ostrinia nubilalis):
O Very good
O Good
O Weak
O Don´t know
O Weak
O Don´t know
2. Pink borer (Sesamia spp):
O Very good
O Good
Year
Event
01Partner
Country
Additional comments:
3.5. Characterise the OTHER pest pressure in MON 810 fields (compared to
conventional maize)
Except the two insects mentioned above, overall assessment of pest susceptibility of
MON 810 compared to conventional maize (insect, mite, nematode pests):
O As usual
O More pests
O Less pests
If the above answer is different from «As usual», please specify the difference in
pest susceptibility in the list and the commentary section below:
1.
2.
3.
4.
5.
6. Other:
O
O
O
O
O
……... O
More
More
More
More
More
More
O
O
O
O
O
O
Less
Less
Less
Less
Less
Less
Additional comments:
3.6. Characterise the weed pressure in MON 810 fields (compared to
conventional maize)
Overall assessment of the weed pressure in MON 810 compared to conventional
maize:
O As usual
O More weeds
O Less weeds
List the three most abundant weeds in your MON 810 maize field:
1.
2.
3.
Year
Event
01Partner
Country
Were there any unusual observations regarding the occurrence of weeds in MON
810 maize?
3.7. Occurrence of wildlife in MON 810 fields (compared to conventional
maize)
General impression of the occurrence of wildlife (mammals, birds, and insects) in
MON 810 compared to conventional maize fields.
O As usual
O More
O Less
O Do not know
If the answer above is «More» or «Less», please specify your observation:
3.8. Feed use of MON 810 (if previous year experience with MON 810)
Did you use the MON 810 harvest for animal feed on your farm?
O Yes
O No
If “Yes”, please give your general impression of the performance of the animals fed
MON 810 compared to animals fed conventional maize.
O As usual
O Different
O Do not know
If the answer above is «Different», please specify your observation:
3.9. Any additional remarks or observations
Year
Event
01Partner
Country
4. Implementation of Bt maize specific measures
4.1. Have you been informed on good agricultural practices for MON 810?
O Yes
O No
Only if you answered “Yes”, would you evaluate these technical sessions as:
O Very useful
O Useful
O Not useful
4.2. Seed
Was the seed bag labelled with accompanying specific documentation indicating
that the product is genetically modified maize MON 810?
O Yes
O No
Did you comply with the label recommendations on seed bags?
O Yes
O No, because
4.3. Prevention of insect resistance
Did you plant a refuge in accordance to the technical guidelines?
O Yes
O No, because the surface of Bt maize planted on the farm is < 5 ha
O No, because
ANNEX 1
Use of existing networks for monitoring MON 810 in
Germany
Kerstin Schmidt, Wenke Mönkemeyer, BioMath GmbH, Schnickmannstrasse 4,
18055 Rostock, Germany
As a basis to identify suitable existing networks for a “German implementation” of the
General Surveillance of MON 810, the German Federal Office of Consumer
protection made available to Monsanto Europe SA a list of 35 potentially usable
networks operating in the field of the environmental monitoring.
Identification of the networks the most relevant to the implementation of MON 810
general surveillance
Federal Water
Monitoring
ÖFS
Nature protection
and species
Biodiversity
Human health
Employment Protection
Water Quality
Sustainability Indicator for
biodiversity
Wildlife
Bees
Songbirds
FFH
Raptors and Owls
Butterflies
Dragonflies
Breeding birds
Environmental Specimen Bank
Food Monitoring
Feed Monitoring/ VDLUFA
Wildlife Feeding behaviour
Bee Poisoning
Animal health
Long term Soil Monitoring
Soil function
Soil Analyses/ VDLUFA
Environmental Specimen Bank
Plant health
PP Service/ISIP
WG Resistances
Sustainable
Agriculture
PPP Use
Environment Indicator
System
Regions
Federal States
Germany
Variety Testing
Operators
networks
Regional Water
Monitoring
Farm Questionnaire, Stewardship Program, Publications
Protection goal
Firstly, the networks were classified in accordance to the protection goals they are
addressing and to their geographical coverage (Figure 1).
Location Registers
Variety Testing
Europe
Geographical extension
Figure 1: Networks (BVL – List) classification by protection goals and geographical coverage.
The core European General Surveillance program (farm questionnaires, stewardship activities,
publication analysis and report from operator networks) are indicated on the right end side.
Secondly, the networks were analysed for their relevance and usability for GMO
monitoring. To that end, a questionnaire was developed and used to interview the
operators of each network about their respective structure and function.
Criteria were established to allow the examination of each network with the view to
using them for the general surveillance of MON 810. These criteria are listed in Table
1.
Table 1 Criteria for the evaluation of existing networks
Criteria
EU or German wide
Explanation
Network covering preferentially a broad base of
European countries, or at least Germany
(methodology applied on a broad basis, consistency
across results)
Cultivation areas of MON 810
Methodology
General
Relevant parameters
Quality
Frequency
Availability
Network that collects data in areas were MON 810 is
cultivated.
Network that possesses valid and transparent
sampling, analysis and reporting methods.
Network that performs “General Surveillance” relevant
to the established protection goals.
Network that collects relevant parameters, with added
value to the farm questionnaire.
High quality data collection (with respect to reliability,
objectivity, topicality , validity, coherence of statistics)
Network is functioning and data are surveyed at least
yearly; data survey is ensured for future years.
Data and/or reports are made available publicly on a
regular basis (at least yearly).
Each criterion was assessed as either being fulfilled (1) or not (0). From the criteria
values an indicator was calculated for the selection of the networks:
 yes, ∑ criteria = 8

indicator = 
no, otherwise

A network is considered as appropriate for the general surveillance of MON 810 if all
criteria are fulfilled.
Based on the indicator values (see ANNEX 2), the following networks were selected
as being suitable to provide information on a relevant monitoring character:
•
•
•
•
Monitoring game species in Germany (WILD)
DDA – Common bird monitoring
Monitoring butterfly population dynamics
Monitoring of bees in Germany
These networks cover especially the protection goal “biodiversity” outside the
agricultural areas.
In addition to these four networks, the indicator values allowed identification of an
additional set of networks considered as suitable to provide information on relevant
influencing factors:
•
•
•
The Environmental core set of indicators;
The Plant Protection Service / ISIP;
Location Registers
Biodiversity
Butterflies
Bees
Breeding birds
Human health
Animal health
Soil function
Plant health
Sustainable
Agriculture
PP Service/ISIP
Environment Indicator
System
Influencing factors
Operators
networks
Monitoring characters
Wildlife
Farm Questionnaire, Stewardship Program, Publications
Protection goal
These networks were identified as being useful to cross check with other data
generated by general surveillance, and more specifically by farm questionnaires
(Figure 2).
Location Registers
Regions
Federal States
Germany
Europe The selected
Figure 2: Networks (Indicator) by protection
goals and geographical
extension.
network are grouped either as monitoring for “monitoring characters” or for “influencing
Geographical extension
factors”
Proposed methodology
After analysing the data and reports of the networks, it was concluded that the most
efficient way of collecting the information was to directly collect the reports published
by the different networks. These reports include a broad level of expert knowledge on
parameters, calculation and interpretation as well as the knowledge of baselines and
thresholds. We therefore propose that Monsanto Europe SA analyse the reports
published annually by the identified networks in view of determining whether MON
810 could potentially have an adverse on the environment.
http://www.jagdnetz.de/Aktuelles/Naturschutz/Informationssystem/
http://www.tagfalter-monitoring.de/
http://www.gesundebienen.de/89/Krankheiten/Bienenmonito
ring/Deutsches_Bienenmonitoring.htm
http://www.dda-birdrace.de/
1
ANNEX 2
SZ Nr. Schutzziel
6 Biodiversität
BP
No.
Beobachtungsprogramm
1 Wildtierinformationssystem der
Länder Deutschlands
6 Biodiversität
2 Brutvogelmonitoring - Monitoring
häufiger Brutvogelarten
6 Biodiversität
3 Brutvogelmonitoring - Monitoring
geschützter und gefährdeter
Brutvogelarten
Beobachtungsgegenstände(soweit bekannt)
Beobachtungsraum
Betreiber / Ansprechpartner
Informationen
Kommentare BVL
Populationsdaten zu den bejagbaren Arten Feldhase,
Rotfuchs, Dachs, Rebhuhn, Aaskrähe
Probenahme jährlich
ausgewählte Referenzgebiete,
bundesweit
Jagdschutz-Verband e.V.
Armin Winter
Johannes-Henry-Str. 26
D-53113 Bonn
Tel 0228 9490631
http://www.jagdnetz.de/communit bundesweit, seit 2000
y/dokumente/download/FlyerWIL
D.pdf
1
Populationsentwicklung weit verbreiteter Brutvogelarten
Brutvögeln, 100 Arten, Wasservögel, Greifvögel,
Singvögel und andere,
Aufnahme je Probenahmeort alle 5 Jahre
1000 Stichprobenflächen à 1km
Größe für bundesweite
Auswertung, unterschiedlich viele
Stichproben für Fragestellungen
auf Länderebene (Σ 2637),
bundesweit
Dachverband Deutscher
Avifaunisten
Johannes Schwarz
Zerbster Str. 7
39264 Steckby
email schwarz@dda-web.de
bundesweit, seit 1989 Daten
fließen ins Pan-European
Common Bird Monitoring ein,
Europa weites Programm mit 36
teilnehmenden Ländern
Bestandsentwicklunh geschützter und gefährdeter
Brutvogelarten
Einzelerhebungen, lokale und
regionale Monitoringprogramme
Dachverband Deutscher
Avifaunisten
Geschäftsstelle
Zerbster Str. 7
39264 Steckby
Tel.: 039244940918
email info@dda-web.de
http://www.tagfaltermonitoring.de/
EU- oder Anbaugebie
Deutschlan
te von
dweit
MON810
Methodik
allgemein
relevante
Parameter
Qualität
Frequenz
Verfügbarke
it
Indikator
Erläuterungen
1
1
1
1
1
1
1
ja
8
1
1
1
1
1
1
1
1
ja
8
Vogelschutzwarten
Erhebungen durch Ehrenamtliche
0
1
1
0
0
1
1
1
nein
5
im Aufbau befindlich, ähnliche
Programme in der Schweiz,
Niederlanden, UK
1
1
1
1
1
1
1
1
ja
8
Nur geschützte Arten,
allgemeiner abgedeckt im
Brutvogelmonitoring (6.2)
6 Biodiversität
4 Tagfalter-Monitoring
Populationsentwicklung von Tagschmetterlinge
durch ErfasserInnen festgelegte
Transekte, bundesweit
UfZ
Department of Community
Ecology
Josef Settele
Theodor-Lieser-Str. 4
06120 Halle
Tel. 0345 5585320
email josef.settele@ufz.de
6 Biodiversität
5 Deutsches Bienenmonitoring
Entwicklung von BienenvölkernKrankheiten,
Rückstandsdaten zu PSM
Ausgewählte Referenzgebiete,
bundesweit
Projektrat des Bienenmonitorings http://www.gesundebienen.de/89/ bundesweit, seit 2005
Krankheiten/Bienenmonitoring/D
Peter Rosenkranz
Landesanstalt für Bienenkunde eutsches_Bienenmonitoring.htm
der Universität Hohenheim (730)
70593 Stuttgart
email bienero@unihohenheim.de
1
1
1
1
1
1
1
1
ja
8
6 Biodiversität
6 Ökologische Flächenstichprobe
Ziel ist eine periodische und bundesweit repräsentative
Datenerfassung zur Struktur von Landschaften und
Biotopen sowie zu deren Artenausstattung als
Grundlage für ein Monitoring Basisvariante:
Biotoptypenkartierung, Blütenpflanzen und Brutvögel
1000 Stichprobenflächen à 1 km²
für bundesweite Auswertungen
Zusätzliche für Auswertungen auf
Landesebene stratifiziert nach
ökologischen Klassen
bundesweite Umsetzung im
Konzept vorgesehen
Bundesländer
0
0
1
1
1
1
1
1
nein
6
Nur ein Bundesland in D, wo
MON810 nicht angebaut wird
1
0
1
1
1
0
0
0
nein
4
Populationsentwicklung von 51
Vogelarten in 6 verschiedenen
Lebensräumen. Allgemeiner
abgedeckt durch
entstanden aus der
Zusammenlegung des Hecken- ,
Höhlenbrüterprogramms und
Singvogelmonitorings Das
Integrierte Singvogelmonitoring
ist im Aufbau z.T. Erhebungen
durch Ehrenamtliche
Monitoring Greifvögel und Eulen http://www.greifvogelmonitoring.d Europa weites Programm in 18
Europas
e/daten.html
EU Staaten, Daten z. T. seit 1957
Schülershof 12
Erhebungen durch
D-06108 Halle / Saale
Ehrenamtliche, in 18 weiteren EUE-Mail: uk.mammen@t-online.de
Mitgliedstaaten
1
1
1
0
0
1
1
1
nein
6
Allgemeiner abgedeckt durch
1
1
1
1
0
1
1
1
nein
7
Daten fließen in Wildmonitoring
(6.1) und in Brutvogelmontoring
(6.2) ein
Bundesländer
0
0
0
1
1
0
0
0
nein
2
Informationen unterschiedlich
von Bundesland zu Bundesland
In NRW: Nutzungstypen, Biotoptypen,
Biotopstrukturen, Brutvögel, Farn- und Blütenpflanzen
in Bezug zu ausgewählten Biotoptypen
6 Biodiversität
7 Nachhaltigkeitsindikator für die
Artenvielfalt
6 Biodiversität
8 IntegriertesSingvogelmonitoring
bestehend aus:
1.Singvogelmonitoring
2.Heckenprogramm
3.Höhlenbrüterprogramm
6 Biodiversität
6 Biodiversität
9 Greifvögel & Eulen Europas
10 Naturschutz- und
http://www3.lanuv.nrw.de
Konzept erstellt, Umsetzung
bisher nur in NRW; Start in SH
In NRW:
Landesamt für Natur, Umwelt
und Verbraucherschutz, NRW
(Landschaftsmonitoring)
In NRW: 120 Stichprobenflächen
à 1 km² stratifiziert nach
Landschaftsräumen
Ziel des Indikators ist es, auf anschauliche Weise den
Zustand von Natur und Landschaft abzubilden
bundesweit
Bundesregierung
1.Beringungsprogramm, langfristige Bestandskontrolle
einheimischer Vogelarten 2.Netzfang und
Revierkartierung zur Erfassung
populationsdynamischer und reproduktionsbiologischer
Aspekte von Heckenvögeln 3. langfristige
Bestandsbeobachtungen von in Höhlen brütenden
Kleinvogelarten verschiedene Singvogelarten
Erfassung von Bestand und /oder Reproduktion einer
oder mehrerer Greifvogel- und/oder Eulenarten
1.Fangplätze in den
Bundesländern 2.Hecken 3.
Nistkästen
Bundesländer, Vogelwarten
Auf durch die ErfasserInnen
bestimmten Kontrollflächen von
mindestens 15 km²
floristische und faunistische Daten
http://www.bundesregierung.de/n
n_1270/Webs/Breg/DE/Politikthe
men/Umwelt/NachhaltigeEntwickl
ung/nachhaltige-entwicklung.html
http://www.bfn.de/0315_nachhalti
gkeit.html
http://www.vogelkundeuntermain.de/hecke.htm
http://www.mu.sachsenanhalt.de/start/wir_ueber_uns/pu
blikationen/files/vogelmonitoringl
sa2003.pdf
Artenmonitoringprogramme der
Länder
6 Biodiversität
11 FFH-Monitoring
Überwachung des Erhaltungszustandes (Monitoring)
der Lebensraumtypen (Anhang I) und Arten (Anhänge
II, IV und V) von europäischem Interesse
FFH-Gebiete, bundesweit
Bundesländer, Koordination BfN http://www.bfn.de/0315_ffh_richtli Stand der Umsetzung in den
nie.html
Bundesländern unterschiedlich,
alle EU-Länder sind verpflichtet
FFH Gebiete zu beobachten
1
1
0
0
0
0
0
0
nein
2
Nicht etabliert.
1
0
1
1
1
1
1
0
nein
6
Informationen unterschiedlich
von Bundesland zu Bundesland
1
1
1
0
1
1
1
0
nein
6
Daten Eigentum des Landwirts,
wird durch Fragebögen
abgedeckt
1
0
0
0
0
1
1
0
nein
3
Proben werden nur genommen,
nicht analysiert
0
0
1
1
0
1
1
0
nein
4
Gewässer durch MON810 Anbau
nicht betroffen insofern nicht
relevant
3 Bodenfunktion
1 Bodendauerbeobachtung
Veränderung und Entwicklung von Böden,
Schadstoffakkumulation
ca. 800 Stichprobenflächen,
Acker, Forst/Wald, Grünland,
Siedlung, Sondernutzung
Bundesländer, UBA
http://www.umweltbundesamt.de/
boden-undaltlasten/boden/bodenschutz/dau
erbeobachtung.htm
3 Bodenfunktion
2 Bodenbeobachtung VDLUFA
Bodenanalysen
Ackerflächen
VDLUFA
c/o LUFA Speyer
Obere Langgasse 40
67346 Speyer
Tel. 06232 136-0
email info@vdlufa.de
http://www.vdlufa.de
3 Bodenfunktion
3 Umweltprobenbank
Probenahme und Archivierung von Umweltproben,
verschiedene Indikatorarten (Pflanzen, Tiere),
Monitoring von Schadstoffen
UBA
http://anubis.uba.de/wwwupb/serv
let/upb
7 Gewässer
1 regionale und örtliche Messnetze
Überwachungsprogramme der Bundesländer
Bundesländer
der Gewässerüberwachung
Light grey area: selected network for relevant monitoring characters
Dark grey area: selected network for relevant influencing factors
Fachgruppen; Lokale
Ansprechpartner;
Auftragsanalytik; keine
allgemeinen Routinestatistiken
Unterschiede hinsichtlich der
gemessenen Parameter und
Messpunktdicht in den Ländern
2
ANNEX 2
7 Gewässer
2 Landesmessnetze
Überwachungsprogramme der Bundesländer
Bundesländer
7 Gewässer
3 Bundesweite
LAWA Messstellen an großen Flüssen
BUND/Länder Arbeitsgemeinschaft Wasser (LAWA),
Bundesländer
Gewässergütekartierung
7 Gewässer
4 PERLODES
Libellen, andere Insekten, Mollusken, andere
Invertebraten, insgesamt 946 Arten
1 Nachhaltige Landwirtschaft
1 NEPTUN
Indizierter Umfang des PSM-Einsatz;
2 Sortenprüfung
Daten aus der Sortenprüfung
3 Nachhaltigkeitsindikator
21 Schlüssel-Indikatoren
7000 Probenahmestellen,
Unterschiede hinsichtlich der
gemessenen Parameter und
Messpunktdicht in den Ländern
http://www.lawa.de
Universität Duisburg-Essen
http://www.uni-duisburgDaniel Hering
essen.de/hydrobiologie/
Universitätsstr. 2
45141 Essen
Tel 021 1833084
email daniel.hering@uni- due.de
seit 2005
0
0
1
1
0
1
1
0
nein
4
relevant
1
0
1
1
0
1
1
1
nein
6
relevant
1
0
0
0
0
0
0
0
nein
1
nicht betroffen
BBA
http://www.bba.bund.de/nn_9210 Nicht kontinuierlich; Erhebungen
32/DE/Home/koordinieren/neptun beschränkt auf bestimmte
/neptun__node.html__nnn=true Kulturen, Zukunft aufgrund
finanzieller Ausstattung unklar
1
0
1
0
1
1
0
0
nein
4
Wissenschaftliches Projekt,
basierend auf
Pflanzenschutzmitteleinsatz
bundesweit
BSA / Antragsteller
http://www.bundessortenamt.de
Fruchtart-Prüfplanabhängige
Aussagen zur Sortenqualität;
Bewertung der gentechnischen
Veränderung in verschiedenen
genetischen Hintergründen
1
1
1
0
1
1
1
1
nein
7
Bewertung von Sorten, nicht von
Events
bundesweit
Bundesregierung
http://www.bundesregierung.de/n
n_1270/Webs/Breg/DE/Politikthe
men/Umwelt/NachhaltigeEntwickl
ung/nachhaltige-entwicklung.html
nur ein Teil der Indikatoren (z.B,
Artenvielfalt, Ernährung) wird für
eine Allgemeine Beobachtung zu
nutzen sein
1
0
0
1
1
0
0
0
nein
3
Nicht etabliert.
http://www.bfn.de/0315_nachhalti
gkeit.html
4 Umwelt - Kernindikatorensystem
Instrument zur Erkennung positiver oder negativer
Entwicklungen in Handlungsfeldern des
Umweltschutzes
bundesweit
UBA
http://www.envevtl. einige der Kernindikatoren
it.de/umweltdaten/public/theme.d für eine GVO-Beobachtung
o?nodeIdent=2702
nutzbar, z.B. Indikatoren zur
Artenvielfalt oder Landwirtschaft
1
1
1
1
1
1
1
1
ja
8
Liste verschiedener Indikatoren.
Insbesondere Daten zu PSM
nützlich für cross check mit
Fragebögen
2 Pflanzenschutz
1 Sortenprüfung
Daten aus der Sortenprüfung
bundesweit
BSA / Antragsteller
http://www.bundessortenamt.de
Fruchtart-Prüfplanabhängige
Aussagen zur Sortenqualität;
Bewertung der gentechnischen
Veränderung in verschiedenen
genetischen Hintergründen
1
1
1
0
1
1
1
1
nein
7
Bewertung von Sorten, nicht von
Events
2 Pflanzenschutz
2 Pflanzenschutzdienste / ISIP
Pflanzen-Schädlinge, Krankheiten;
Bundesländer
Pflanzenschutzdienste der
Länder
http://www.isip.de
ISIP: Beschränkt auf best.
Feldfrüchte und Schädlinge; z.Z.
auf Beratung der Landwirte
fokussiertPSD:
Regional/länderspezifisch
organisiert; Beraternetzwerke
1
1
1
1
1
1
1
1
ja
8
2 Pflanzenschutz
3 AG Resistenzen
Schädlingsresistenzen: Fungizide, Herbizide,
Insektizide, Rodentizide
BBA; DPG
http://www.bba.bund.de/cln_045/ Informelle Experten-Netzwerke
nn_921062/DE/Home/pflanzen__ unter Beteiligung verschiedener
schuetzen/pfsmittel/resistenz__p Institutionen.(Im Aufbau)
sm/resistenz__psm__node.html_
_nnn=true;http://p11631.typo3ser
ver.info/herbizidresistenz_u.html
1
0
0
0
1
0
0
0
nein
2
4 Tiergesundheit
1 Überwachung der Futtermittel
1
1
1
0
1
1
1
0
nein
6
Daten Eigentum des Landwirts,
wird durch Fragebögen
abgedeckt
4 Tiergesundheit
2 Jagdwissenschaftliche Institute,
1
0
0
0
0
0
0
0
nein
1
Daten fließen in Wildmonitoring
(6.1) ein
Institut für Wildtierforschung
Privatwirtschaftl. QM; Labore der http://www.vdlufa.de
VDLUFA
Fachgruppen; Lokale
Ansprechpartner;
Auftragsanalytik; keine
allgemeinen Routinestatistiken
Toxikologische Wirkungen durch verändertes
Fraßverhalten bei Wildtieren (Dachs, Hase, Reh)
4 Tiergesundheit
3 Bienenvergiftungen
Analyse von Bienenvergiftungen (PSM)
5 Menschliche Gesundheit
1 Umweltprobenbank
Probenahme und Archivierung von Humanproben
2 ggf. Landesämter für
wenige Probenahmepunkte
BBAUntersuchungsstelle für
Bienenvergiftungen
Messeweg 11 / 12
38104 Braunschweig
http://www.bba.bund.de/cln_044/
nn_1003268/DE/Home/pflanzen_
_schuetzen/bienen/bienen__nod
e.html__nnn=true
1
1
1
1
1
1
1
1
ja
8
Funktionierendes Alarmsystem,
Berichte fließen in 6.5 ein
UBA
http://anubis.uba.de/wwwupb/serv
let/upb
1
0
0
0
0
1
1
0
nein
3
Proben werden nur genommen,
nicht analysiert, ferner abgedeckt
durch GVO Import
1
0
0
1
0
0
0
0
nein
2
Abgedeckt durch GVO Import
1
0
1
1
0
0
0
0
nein
3
in Entwicklung
1
0
1
1
1
1
1
1
nein
7
Abgedeckt durch GVO Import
Arbeitsschutz/ Landwirtschaftliche
Berufsgenossen-schaften/
Arbeitsmedizinische Dienstes
3 Human-Biomonitoring
4 Lebensmittel-Monitoring
GSF-Forschungszentrum für
Umwelt und Gesundheit
http://www.gsf.de/infostellehumanbiomonitoring/index.php
Anteil von GVO in Lebensmittel sowie Verstöße gegen bundesweit
geltendes Recht vorliegen
BVL
www.bvl.bund.de
www.bvl.bund.de
ohne direkte Verbundung zu
Schutzgütern
Standortregister
Lage von GVO Flächen
bundesweit
BVL
Schutzgütern
Bodennutzungshaupterhebung
Flächenerhebungen nach Art der tatsächlichen
Nutzung
bundesweit
Bundesländer, Statistisches
Bundesamt
Schutzgütern
EU-MON
europaweit (EU)
UFZ
Department of Conservation
Biology
Klaus Henle
Permoser Str. 15
04318 Leipzig
Light grey area: selected network for relevant monitoring characters
Dark grey area: selected network for relevant influencing factors
http://eumon.ckff.si/index1.php
Im Aufbau
kein Beobachtungsprogramm per
se, aber eine Erhebung welche
Beobachtungsprogramme in
Europa durchgeführt werden, als
zusätzliche Quelle für
Informationen evtl. geeigneter
Beobachtungsprogramme

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