W. Müller,

EFSA European Food SCANDAL AuthorityEuropean Food SCANDAL Authority

1www.foeeurope.org www.Global2000.atwww.eco-risk.at

Legal RequirementsA i l t ff t EC R 1829/2003 Assessing long- term effects EC Reg. 1829/2003

in conjunction with178/2002 Article 14

Directive 2001/18/EC Annex II

Assessing delayed effects and indirect effects

Directive 2001/18/EC Annex II

Assessing cumulative long term effects on human health, soil fertiliy, flora

EC Decision Description of uncertainties

effects on human health, soil fertiliy, flora fauna

2002/623p

e.g. assumptions made in the risk assessment, and of the known limits of mitigation measuresmitigation measures

Crop approved (2001/18/EC) Test• Ms8 x Rf3 (2005) • no toxicol. test(2001/18/EC) Test

• Bt 11 cult. (2005)

• 1507 culti. (2005)

• no toxicol. test• 90 days subchronic1507 culti. (2005)

• Mon 863 (2004)

GT 73 (2004)

• 90 days subchronic

• no toxicol test• GT 73 (2004)

• NK 603 (2003)

• no toxicol. test• 90 days subchronic( )

GMO are approved without any chronicGMO are approved without any chronic toxicol. test = long term = 720 days study

Tests on Pesticides vs GMOs

GMOPesticide

Tests on Pesticides vs GMOs

yesAssessing long- term effects chronic tox study (720 days)

no

yesAssessing effects on subsequent generations

tox. study (720 days)

no

noAssessing cumulative toxic effects

generationsno

noDescription of uncertaintiese.g.assumptions made in the risk

no

g passessment, and of the known limits of mitigation measures

EFSA methodsEFSA methodsMethod Comment• Comparative chemical

analyses of protein, amino acid content, ash content

• No scientific basis of how to translate results into human toxicity assessment,

etc.• Sequence Analyses • Almost identical sequences can show

differences in function monkey/human• 28 days study with the

protein

differences in function monkey/human DNA

• Short term toxic studies are useless, d b id d f fand must be avoided from terms of

animal rights• Comparative 90 day study • Subchronic study, not able to

5

Comparative 90 day study with rats ( NK603 and Mon863 but not in GT73)

Subchronic study, not able to extrapolate to chronic effects (cancergogenicity, immuno toxicity)

EFSA vocabulary on observed statistically f d ff b d lsignificant differences between GM and control

phrases sourcephrases source1. Altered level of linolenic acid is

considered as not biologicallyi ifi t

Rape GT 73 (Monsanto) EFSA Journal 2004, 29:1-19significant, greater differences

between GT73 and Westar but without statistical analyses

19

1. no consistent differences, 2. no biological significance,3. artifactual differences of

Maize NK 603(Monsanto) EFSA Journal 2003, 9:1-143. artifactual differences of

corbuscular haemoglobin values(90 days feeding study)

4 No conclusive differences of

,

6

4. No conclusive differences ofchemical constituents

EFSA EFSA

• Up to now ALL observed differences Up to now ALL observed differences between GM and Non-GM variety had beentolerated by EFSA.y

• No argumentation to what extent observed differences are generally tolerated is g yprovided.

• The question remains why these parameters q y pare tested when statistically significant differences are not of biological relevance.

7

Wording of Monsanto and EFSA e g NK603Wording of Monsanto and EFSA e.g. NK603Data interpretation of Judgement by

MonsantoJudgement by EFSA

Monsanto

observed differences found in the subchronic 90 days

absence of biologically relevant

“The applicant concludes that these findings are of no biological significance The subchronic 90 days

toxicity studyrelevant differences

biological significance. The panel accepts this as a reasonable interpretation of the data ”the data.

safety claims of CP4 EPSPS-Protein

the long history of safe

humans have a long history of dietary exposure to the protein. y

consumption of similar proteins

y p pNo adverse effects associated with its intake have been identified.

8

Is it sound science to say GMOs yare safe?

No!

What do we know what we don´t know

10

synthetic gene new for humans

Mon810 maize- YieldGardTM

(M )

synthetic gene new for humans

(Monsanto)Maize DNA P-35S hsp70 intron CryIA(b) T noshsp70 intron CryIA(b)

Virus Bt-Bacteria - Soil-

T-nos

maize a e atruncated

SoBacteria

Synthetic genes are man made genes and do not exist in

11any natural living species on the planet

http://www.geneart.com GC content cryptic splice sites premature –... GC content, cryptic splice sites, premature

poly(A) sites, AT rich killer sequences, RNA secondary structures, structures,

– often makes it necessary to adapt and optimize the gene of interest towards the genetic the gene of interest towards the genetic requirements of the host organism.

–Apparently, an optimized sequence does not ppa e t y, a opt ed seque ce does otoccur in nature and has to be designed on a rational basis followed by in vitro synthesis.

–GENEART offers both steps:

●state-of-the-art gene optimization plusg p p

● fast and reliable de novo gene synthesis.

Partikel Gun

Gene Gun 1 of 1000 sucessful integration

Grafik: BioLinX GmbH

synthetic genes cause unintended recombinationssynthetic genes cause unintended recombinations

CHARACTERISATION OF COMMERCIAL GMO INSERTS: A SOURCE OF USEFUL MATERIAL TO STUDY GENOME FLUIDITY.

T25 maize - LibertylinkTM (Bayer)Tolerance to herbicide glufosinate, Peg-mediated transformation

Construct content : truncated bla gene (bla*), pUC cloning vector (pUC), synthetic pat gene (pat), CaMV 35S promotor and terminator (P35S, T35S).

Sequence expected

DNA rearrangement: presence of a second truncated and rearranged P35S on the 5’ end.

Maize DNAP35S* pUC18 P35S pat T35S pUC18

bla*

bla*

pUC18 P35S pat T35S

bla*

Sequence observed

Sequence expected(public data)

(Presence of cloning vector + the 5 first bp of bla on the 3’ end )

Mon810 maize- YieldGardTM (Monsanto)Resistance to lepidopteran insects Bombardment

(Collonnier et al. (2003) Eur. Food Res. Tech. (submitted))

g p g

Insertion site: the 5’ and 3’ ends of the insert show homologies with Huck retrotransposons.

Mon810 maize YieldGard (Monsanto)Resistance to lepidopteran insects, BombardmentConstruct content : CaMV 35S promotor (P35S), CryIA(b) toxin synthetic gene (CryIA(b)), nos terminator (T-nos).

DNA d l ti f T i th i t (b t T d t t d i th ) d d l ti f t

Sequence observedSequence expected Maize DNA

P-35S hsp70 intron CryIA(b) T-nos

P-35S hsp70 intron Truncated CryIA(b)Maize DNA

14(Hernandez et al. (2003) Transgenic Res. 12: 179-189; Holck et al. (2002) Eur. Food Res. Tech. 214: 449-453)

DNA rearrangement: deletion of T-nos in the insert (but Tnos detected in the genome) and deletion of a partof CryIA(b).

Insertion site: the 5’ end of the insert shows homology with LTR sequences of the Z. mays alpha Zein genecluster. No homology between LTR sequences and the 3’ end: rearrangement of the integration site.

Gold in the gene-desert (junk DNA) Science (2003) 302:41Non-coding genes/(RNA genes)

15

Non coding genes/(RNA genes)

Eric Neumann, vice president of bioinformatics at Beyond Genomicsvice president of bioinformatics at Beyond Genomics

” W ll h d di f h” We really have a poor understanding of what a gene actually does and where and when it g yshould do it. You can understand the entire genome and [still] understand less than 1genome and [still] understand less than 1 percent about what is going on in a cell." DODGE J (2003) Data glut. The Boston GlobeDODGE J (2003) Data glut. The Boston Globe

http://www.boston.com/

16

Food-DNA pieces of rubisco gene had been detected in lymphocytes blood liver detected in lymphocytes, blood, liver,

spleen, kidney, muscles and milk

17

18

Synthetic DNA IN BLOOD AND TISSUESy• Mazza R, Soave M, Morlacchini M, Piva G, Marocco A

(2005) Assessing the transfer of genetically modified DNA(2005) Assessing the transfer of genetically modified DNA from feed to animal tissues. Transgenic Research 14: 775-784.

• Sharma R, Damgaard D, Alexander TW, Dugan ME, Aalhus JL Stanford K McAllister TA 2006 Detection ofAalhus JL, Stanford K, McAllister TA. 2006 Detection of transgenic and endogenous plant DNA in digesta and tissues of sheep and pigs fed Roundup Ready canola meal.J Agric Food Chem. 2006 Mar 8;54(5):1699-709.

Several receptors for foreign DNA/RNASeveral receptors for foreign DNA/RNA

20

Food-DNA pieces interact directly with the iimmune system

• The protective effects of probiotics are mediated by their own DNA rather than by their by their own DNA rather than by their metabolites or ability to colonize the colon

– Rachmilewitz et al: Gastroenterology 2004 Feb;126(2):520-8Feb;126(2):520 8

21

transgenic pea causes allergenic i reactions

PRSCOTT V 2005 T i E i f B A lPRSCOTT V 2005: Transgenic Expression of Bean r-Amylase Inhibitor in PeasResults in Altered Structure and Immunogenicity: J. Agric. Food Chem. 2005, 53, 9023−9030 9023

With methods used by EFSA this transgenic pea would have been

approved save.ppSo far all GMO applications has been

pp o ed e b EFSA approved save by EFSA.

short list of effects of GMO from studies of the biotech industry von Kurzzeit-Studien the biotech industry von Kurzzeit Studien

ignored by EFSAsourceeffects90 days study

Mais Mon 863 (Monsanto), EFSA Journal 2004, 50:1-25

1. increase counts lymphocytes, differences in kidney cells

2 diff f bl d

sourceeffects90 days study

Mais NK 603 (Monsanto), EFSA Journal 2003, 50:1-25

R GT 73 (M t )

2. differences of blood parameters(artifactual differences“ of

corbuscular haemoglobin values)Raps GT 73 (Monsanto), EFSA Journal 2003, 50:1-25

Mais 1507 (Bayer)

3. 15- 16 % increased liver weights

4. changes of liver, kidney and blood EFSA Journal 2004, 124:1-18

parameters

”While the duty of preventing damage to the environment is based on a known the environment is based on a known

risk, the notion of precaution is based on lack of certainty.”(OECD 2001)lack of certainty. (OECD 2001)

as a consequence of the lack of long-term tests and major uncertainties in the risk assessement of

GMOs the approval of GMOs is not in line with the

precautionary principle as outlined in Directive2001/18 and Regulation 1829/2003

27

Summary Summary

• We eat synthetic genesWe eat synthetic genes• We know almost nothing about genes

d j k und junk genes• Pieces of food-DNA are found in Pieces of food DNA are found in

various organsF d DNA i t t ith th i • Food-DNA interacts with the immune system

• Transgenic crops are badly tested

Why we are forced to eat GM yfood?

EFSA pushes for GMOs instead of scientific evaluation of scientific evaluation

Solution?Solution?

Change the approval system –Change the approval system

don´t allow EFSA to play a don t allow EFSA to play a

decisive role in the approval decisive role in the approval

systemy