Statement of EFSA: Statement on an emergency measure prohibiting the cultivation of maize MON 810

An ongoing stand-off: French politics versus science:

On 25 March 2014, the European Commission requested the European Food Safety Authority (EFSA) to assess the supporting documentation submitted by France to notify an emergency measure under Article 34 of Regulation (EC) 1829/2003, prohibiting the cultivation of genetically modified maize MON 810 in the EU.

EFSA assessed the documentation supplied by the French Authorities and the scientific publications cited in the French Authorities’ report. For each area of concern outlined in the French Authorities’ report, EFSA assessed whether any of the scientific publications not previously addressed by EFSA and/or its GMO Panel, or any of the arguments put forward by France, would invalidate the previous GMO Panel conclusions on the safety of maize MON 810. Moreover, EFSA considered the relevance of concerns raised by France in the light of the most recent and relevant scientific data published in the scientific literature.

During its evaluation of the French Authorities’ report, EFSA noted that most of the cited scientific publications were addressed previously by EFSA and its GMO Panel in various scientific outputs. These publications were therefore not considered further. For the remaining scientific publications, EFSA has focused its assessment on those aspects that are relevant to maize MON 810.

Neither the scientific publications cited in the French Authorities’ report with relevance to maize MON 810 nor the arguments put forward by France reveal any new information that would invalidate the previous risk assessment conclusions and risk management recommendations made by the EFSA GMO Panel. Therefore, EFSA considers that the previous GMO Panel risk assessment conclusions and risk management recommendations on maize MON 810 remain valid and applicable.

EFSA concludes that, based on the documentation submitted by France, there is no specific scientific evidence, in terms of risk to human and animal health or the environment, that would support the adoption of an emergency measure on the cultivation of maize MON 810 under Article 34 of Regulation (EC) 1829/2003.

Keywords
GMO, maize (Zea mays), MON 810, France, emergency measure, environment, Directive 2002/53/EC

BACKGROUND AS PROVIDED BY THE EUROPEAN COMMISSION AND EFSA

The marketing of maize MON 810 (notification C/F/95/12-02) was authorised under Directive 90/220/EEC in the European Union (EU) for all, other than food, uses by the Commission Decision 98/294/EC of 22 April 1998 (EC, 1998). Consent was granted to the applicant (Monsanto Europe S.A.) by France on 3 August 1998. Food uses of maize derivatives were notified according to Article 5 of the Novel Food Regulation (EC) No 258/97 on 6 February 1998.

On 15 June 2009, the EFSA GMO Panel issued a scientific opinion on the renewal of the authorisation for the continued marketing of: (1) existing food and food ingredients produced from maize MON 810; (2) feed consisting of and/or containing maize MON 810, including the use of seed for cultivation; and (3) food and feed additives, and feed materials produced from maize MON 810. The EFSA GMO Panel concluded that “maize MON 810 is as safe as its conventional counterpart with respect to potential effects on human and animal health”, and that “maize MON 810 is unlikely to have any adverse effect on the environment in the context of its intended uses, especially if appropriate management measures are put in place in order to mitigate possible exposure of non-target (NT) Lepidoptera”. The EFSA GMO Panel recommended that “especially in areas of abundance of nontarget Lepidoptera populations, the adoption of the cultivation of maize MON 810 be accompanied by management measures in order to mitigate the possible exposure of these species to maize MON 810 pollen”. In addition, the EFSA GMO Panel advised that “resistance management strategies continue to be employed and that the evolution of resistance in lepidopteran target pests continues to be monitored, in order to detect potential changes in resistance levels in pest populations” (EFSA, 2009).

On 30 November 2011, the EFSA GMO Panel adopted a statement supplementing the environmental risk assessment conclusions and risk management recommendations on maize Bt11 cultivation (EFSA Panel on Genetically Modified Organisms, 2011c). In its statement, the EFSA GMO Panel concluded that “subject to appropriate management measures, maize Bt11 cultivation is unlikely to raise additional safety concerns for the environment compared to conventional maize” (EFSA Panel on Genetically Modified Organisms, 2011c). The EFSA GMO Panel considered that the environmental risk assessment conclusions and risk management recommendations on non-target Lepidoptera for maize Bt11 apply equally to maize MON 810 due to the similarities between both Bt-maize events (i.e., identity of amino acid sequence of the core of the Cry1Ab protein, similar biological activity against susceptible Lepidoptera, similar Cry1Ab protein expression level in pollen).

The EFSA GMO Panel further supplemented its previous risk management recommendations on maize Bt11 and MON 810 cultivation by reapplying the mathematical model developed by Perry et al. (2010, 2011, 2012), in order to consider additional hypothetical agricultural conditions, and to provide additional information on the factors affecting the insect resistance management (IRM) strategy (EFSA Panel on Genetically Modified Organisms, 2012d).

On 6 December 2012, following a request from the European Commission, the EFSA GMO Panel compiled its previous risk assessment conclusions and risk management recommendations on maize MON 810, and considered their validity in the light of new relevant scientific publications published from 2009 onwards (EFSA Panel on Genetically Modified Organisms, 2012e). Based on the performed literature search, the EFSA GMO Panel concluded that “its previous risk assessment conclusions on maize MON 810 as well as its recommendations on risk management measures and monitoring remain valid and applicable.”

Following requests of the European Commission to assess the annual post-market environmental monitoring reports on maize MON 810 cultivation submitted by the applicant, the EFSA GMO Panel issued scientific opinions on the 2009, 2010, 2011 and 2012 PMEM reports on maize MON 810 (EFSA Panel on Genetically Modified Organisms, 2011a, 2012a, 2013c, 2014). The EFSA GMO Panel noted shortcomings in the methodology for case-specific monitoring and general surveillance, and made recommendations to strengthen the annual PMEM activities on maize MON 810. So far, the Several EU Member States invoked safeguard clause or emergency measures to provisionally restrict or prohibit the marketing of maize MON 810 on their territory.

For all cases for which the EFSA GMO Panel or EFSA has been asked by the European Commission to evaluate whether the invocation was justifiable on the basis of the scientific information submitted in support of a safeguard clause or emergency measures, the EFSA GMO Panel or EFSA concluded that, in terms of risk to human and animal health and the environment, no new scientific evidence had been presented that would invalidate its previous risk assessment conclusions on maize MON 810 (EFSA, 2004, 2005, 2006a, b, 2008a, b, c, d, 2014a; EFSA Panel on Genetically Modified Organisms, 2012b, c, 2013a, b). ...

More @ Statement of EFSA: Statement on an emergency measure prohibiting the cultivation of maize MON 810:
(Full document pdf EFSA Journal 2014;12(8):3809 European Food Safety Authority)
Quotes of Cry protein safety issues raised:
ASSESSMENT 
At the request of the European Commission, EFSA assessed the documentation supplied by France (referred to hereafter as the French Authorities’ report) and the scientific publications cited in the French Authorities’ report. For each area of concern outlined in the French Authorities’ report, EFSA assessed whether any of the scientific publications not previously addressed by EFSA and/or its GMO Panel, or any of the arguments put forward by France, would invalidate the previous GMO Panel conclusions on the safety of maize MON 810. Moreover, EFSA considered the relevance of concerns raised by the French Authorities in the light of the most recent and relevant scientific data published in the scientific literature.
During its evaluation of the French Authorities’ report, EFSA noted that most of the cited scientific publications were addressed previously by EFSA and its GMO Panel in various scientific outputs (e.g., EFSA Panel on Genetically Modified Organisms, 2011b, c, 2012d, 2013c; EFSA, 2014b). These publications are therefore not considered further here, except for HCB (2013), which formed a key part of the basis of the scientific argumentation put forward in the French Authorities’ report.

In the case of the remaining scientific publications (Campagne et al., 2013; Mezzomo et al., 2013; Zhou et al., 2014), EFSA has focused its assessment on those aspects that are relevant to maize MON 810.

The EFSA assessment below is structured into the Sections used in the French Authorities’ report. ...

2. EFSA assessment of Section I.3, “Impact of MON810 maize on non-target invertebrates” as referred to in the French Authorities’ report 
2.1. Cited scientific publications 
From the scientific publications in the French Authorities’ report with relevance to maize MON 810, two publications (Mezzomo et al., 2013; Zhou et al., 2014) were not previously considered by EFSA and/or its GMO Panel. The possible relevance of these publications for the risk assessment of maize MON 810 was scrutinised.

- Mezzomo et al. (2013): Mezzomo et al. (2013) studied the potential haematotoxicity (by conventional haematology) and genotoxicity (by in vivo micronucleus test) of spores of four Bt-strains expressing Cry toxins in Swiss albino mice. These Bt-strains were genetically modified to express Cry1Aa, Cry1Ab, Cry1Ac or Cry2Aa. The spores, re-suspended in distilled water, were given by oral gavage in a single administration of one of three doses (27, 136 or 270 mg of lyophilised spores/kg body weight (bw)). Binary combinations of lyophilised spores were also given at 270 mg/kg bw. The negative control group received distilled water (vehicle). As a positive control for the micronucleus test, mice received cyclophosphamide at 27 mg/kg bw. Group size was three male and three female mice. Blood and bone marrow were sampled after 24 hours in all groups. Blood samples were also taken after 72 hours or 7 days from additional groups receiving the high dose of the individual Bt-strains or 27 mg cyclophosphamide/kg bw. Based on the results, the authors concluded that administrations of Bt-spores provoked selective haematotoxicity, particularly for the erythroid lineage; a significant reduction in bone marrow cell proliferation demonstrated cytotoxic but not genotoxic effects.
- Zhou et al. (2014): Zhou et al. (2014) reported that the Cry1Ab toxin affects the metabolic enzymes acetylcholine esterase, glutathione peroxidase and superoxide dismutase in the predatory spiders Ummeliata insecticeps and Pardosa pseudoannulata when fed Cry1Ab-containing prey (fruit flies). The authors concluded that plant-produced Bt-proteins can affect non-target arthropods at the physiological and biochemical level, and reduce their fitness. 

2.2. Relevance of the cited publications for the risk assessment of maize MON 810 
2.2.1. Haematotoxicity (Mezzomo et al., 2013)
Mezzomo BP, Miranda-Vilela AL, Freire IdS, Barbosa LCP, Portilho FA, Lacava ZGM and Grisolia CK, 2013. Hematotoxicity of Bacillus thuringiensis as spore-crystal strains Cry1Aa, Cry1Ab, Cry1Ac or Cry2Aa in Swiss albino mice. Journal of Hematology & Thromboembolic Diseases, 1, 104.

The test items were not purified Cry toxins, but viable Bt-spores expressing the Cry toxins. In addition, the Bacillus species used is known to be capable of producing enterotoxins (e.g., Gaviria Rivera et al., 2000) and a range of other toxic materials (Butko, 2003). Therefore, in the absence of an appropriate Bt-control strain, it is not possible to attribute any observed findings specifically to the Cry toxins, as was done by Mezzomo et al. (2013) in describing their results.
This acute study showed several key weaknesses which impede interpretation of the results. First, a study which addresses only peripheral blood findings of haematotoxicity has limited value in the absence of gross morphological and/or histopathological investigations. Haematological changes may be secondary to other pathological conditions (for example to haemorrhages) which were not monitored in this study. Second, vehicle control groups for the evaluation of peripheral blood analyses at 72 hours and 7 days after administration were missing. Finally, there is a lack of data indicating normal variation of the measured parameters under the study conditions, particularly considering the small number of animals per sex and per group and the use of a single vehicle control group (at 24 hours after administration). In addition, the micronucleus test was not performed in accordance with the current standards (OECD Guideline TG 474), in particular with respect to the number of animals used (only three animals per sex and group, instead of five), the dosing regime (sampling at only one time point, instead of at least two), as well as the presentation and interpretation of results (only group data but not individual data were considered).
Regarding the endpoints measured 24 hours after administration, the authors reported that four parameters of the erythrogram (mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV) and red cell distribution width (RDW)) showed statistically significant differences compared with the vehicle control group, but no significant differences were identified in the leucogram. Overall, EFSA considers that the values of the erythrogram parameters for the four Bt-strains and for the three dose levels are very similar. The only notable difference remained between the test groups and vehicle control group for the MCV and its coefficient of variation (RDW). These differences were small in magnitude (around 10 %) and were not dose-related. Furthermore, these parameters are derived by calculation from the haematocrit, red blood cells (RBCs) and haemoglobin content of the blood, none of which showed a significant difference. Decreases in the percentage of polychromatic erythrocytes in the bone marrow of treated animals relative to the vehicle control group were also used as an argument to support effects on the erythroid lineage by the authors. However, owing to the limitations of the micronucleus test performed, this conclusion is not justified.
The authors also reported significant differences in platelet counts and related parameters (mean platelet volume (MPV), platelet large cell ratio (P-LCR) and platelet distribution width (PDW)) measured 24 hours after administration. Platelet counts are known to be highly variable, and the corresponding figures for the three dose levels showed no evidence of dose-effect relationship.
Based on these findings, EFSA concludes that the Mezzomo et al. (2013) publication does not support the conclusions on haematotoxicity associated with the Cry toxins.

2.2.2. Potential adverse effects on spiders (Zhou et al., 2014) 
Zhou J, Xiao K, Wei B, Wang Z, Tian Y, Tian Y and Song Q, 2014. Bioaccumulation of Cry1Ab protein from an herbivore reduces anti-oxidant enzyme activities in two spider species. PLoS ONE, 9, e84724.

The findings reported by Zhou et al. (2014) suggest that the Cry1Ab toxin can affect enzyme activity in predatory spiders when fed Cry1Ab-containing prey. Even though the spiders have been shown to take up the Cry1Ab toxin when feeding on the Cry1Ab-containing prey, no correlation can be made between the Cry1Ab toxin content measured in the spiders and the observed differences in enzyme activity in the spiders. Some limitations of the study (such as undefined number of organisms tested and replications; lack of details on how the protoxin was quantified in the diet; ill-characterised and -described purity of the Cry1Ab toxin; lack of a positive control) make it difficult to explain the mechanism leading to the observed differences. Therefore, EFSA considers that the dataset reported by Zhou et al. (2014) is preliminary and is not sufficient to demonstrate a new mode of action of the Cry1Ab protein on spiders.
In addition, the biological relevance of the observations made remains difficult to assess, as it is not clear how the observed differences in enzyme activity are related to fitness parameters of the spiders; the authors did not report on life-table parameters of the spiders. Moreover, the reported response was transient.
Finally, exposure has not been sufficiently characterised. The test model used by Zhou et al. (2014) does not reflect realistic exposure conditions; spiders are generalist predators, and the level of exposure to the Cry1Ab toxin will depend on the prey spectrum of individual species. In the experiments performed by Zhou et al. (2014), the variability of food uptake was not controlled, and this may have influenced the observed variability of the Cry1Ab toxin content in the spiders. Therefore, it remains challenging to extrapolate the results of the observations made under controlled conditions to spider activity under field conditions.
At present, EFSA is not aware of identified significant adverse effects of the Cry1Ab toxin on spiders. Laboratory studies have indicated that plant-produced Cry proteins have no direct effects on life-table parameters of spiders after ingestion, whereas field studies confirmed that population densities of spiders are not adversely affected (reviewed by Peterson et al., 2011). Those findings are in line with the outcomes of meta-analyses, in which a broad range of beneficial arthropods including spiders were addressed (Marvier et al., 2007; Wolfenbarger et al., 2008; Naranjo, 2009; Peterson et al., 2011; Albajes et al., 2013; Comas et al., 2014).

2.3. EFSA conclusion 
Neither the results reported by Mezzomo et al. (2013) and Zhou et al. (2014) nor the arguments put forward by France in the French Authorities’ report reveal any new information that would invalidate the previous risk assessment conclusions and risk management recommendations made by the EFSA GMO Panel.