Colloquium on GM for NGOs & Civil Society Organisation Sheepdrove Trust & Roddick Foundation

1. Colloquium on GM for NGOs & Civil Society Organisation Sheepdrove Trust & Roddick Foundation Sources and Mechanisms of Health Risks from Genetically Modified Crops and Foods Sheepdrove, 26th Feb 2013 Dr Michael Antoniou

2. Earth Open Source www.earthopensource.org

3. GM Food Safety: should we be worried? • Genetically modified (GM) crops are produced through a procedure that is radically different from conventional plant breeding: • New technology: only ~40 years old. • Does not involve natural sexual reproduction methods. • Allows transfer of one or few genes between totally unrelated organisms,employing artificial combinations of genetic material. • Crosses species barriers to reproduction in ways that do not occur naturally • Produces combinations of genes that have not evolved to work together in a coordinated integrated whole • GM transformation process is inefficient: frequently uses antibiotic resistance genes to select for transformants, which can persist in final crop.

4. GM Food Safety: why should we be worried? • Generation of GM plants (and animals) involves the random insertion or splicing of a foreign gene into the host DNA/genome and is not a “clean” process. • GM transformation process as a whole - transgene insertion plus tissue culture - is highly mutagenic: GM to a lesser or greater degree, disrupts host genetic order and function. • Combined effects of GM (mutation effects; novel combinations of gene products): • can disrupt genetic and protein biochemical function • lead to the generation of novel toxic effects, allergies and altered nutritional value. • Therefore, GM crops pose new risks to food safety that need to be evaluated for both acute and especially chronic toxic effects.

5. There are three sources of health risks that can potentially arise from GM foods: • 1. The introduced foreign GM gene (“transgene”): • GM gene product directly (e.g. Bt toxin) • Altered plant biochemistry caused by GM gene product (e.g. enzymes conferring herbicide tolerance) • 2. Higher exposures to herbicides used in conjunction with the cultivation of GM crops (e.g. glyphosate) • 3. Altered plant biochemistry caused by mutagenic effect of the GM transformation process

6. Only four crops and two traits dominate GM based agriculture: • Soy beans • Maize or corn • Canola or oilseed rape • Cotton • All the GM soy and some corn varieties are engineered to be tolerant to glyphosate based herbicide application (mostly Roundup formulations) • Most corn and cotton are predominantly engineered to express versions of the insecticidal Bt toxin protein; some combine both Bt toxin and glyphosate tolerance • Cultivation restricted to a few nations: N & S America

7. Health Risk Evaluation of GM FoodSubstantial Equivalence, Generally Recognised as Safe and “Comparative Assessment” General biochemical analysis only. Assessment ofknowntoxins/allergens only. GM and non-GM parental plant are “substantially equivalent” if they contain similar amounts of biochemical components within limits of natural variation. No feeding trials formally required if substantial equivalence is found. FLAWS: Only looks at gross biochemical composition; only looks at known components. NOTE:BSE cow is “substantially equivalent” to a normal cow!

8. Animal feeding toxicity tests are not mandatory anywhere in the world • Organisation for Economic Co-operation and Development (OECD): • Sets guidelines for industry to conduct animal feeding studies to evaluate toxicity of chemical products (e.g. herbicides, insecticides) • Multiple doses (at least 3 to determine dose response effects) • More than one animal • Only 90 days duration • Has not issued guidelines for GM feed/food toxicity feeding studies • However: • Industry has adopted the chemical toxicity OECD guidelines to GM feed/food animal feeding studies • Applications for marketing within the EU • [Note: not all OECD guidelines are followed by industry; e.g., use of at least 3 doses, use of multiple animals species].

9. Heath Risk Evaluation of GM Foods • Determining the effect of the GM transformation process: • Compositional • Animal toxicity feeding studies • Need to minimise variables • Industry’s approach: • compare GM crop / food with large number of varieties unrelated to • GM variety under study grown at different times and locations • Use data in the literature (“historical norms”) • Outcome: • increases variables or “noise” in the system • masks rather than highlights effect of GM process • Only scientifically valid comparators: GM vs non-GM • parent (“isogenic”) grown at same time and location

10. Industry GM Crop Applications to EU For example: MON863, MON810, NK603 corn GM substantially equivalent to non-GM 90 day feeding studies in rats Multiple non-isogenic non-GM comparators (note: goes against EU guidelines) Statistically significant differences in organ function (e.g. liver, kidney, blood system) between GM and non-GM not significant: Fell within large range of variation caused by using non-isogenic controls EU regulators accepted these arguments; passed all as safe for consumption

11. Controlled Animal Feeding Studies Show Clear Signs of Toxicity linked with GM cropsRevealed by GM vs isogenic non-GM comparison

12. Feeding studies conducted by industry • Rats fed commercialised insecticide-producing MON863 Bt corn: • Grew more slowly • Sex differences • Showed higher levels of certain fats (triglycerides) in their blood • Problems with liver and kidney function (Séraliniet al., 2007). Note: * & ** indicate statistical significance

13. Feeding studies conducted by industry Rats fed commercialised GM Bt corn varieties MON863 and MON810 and Roundup tolerant NK603:signs of toxic effects on liver and kidneys. (de Vendomois et al., 2009). Differences in NK603 fed rats and control animals fed isogenic non-GM maize. Note: * & ** indicate statistical significance

14. Heath Risk Evaluation of GM Crops Industry position and accepted by regulators: Accepts statistically significant differences between GM and non-GM feeding groups But claims not biologically significant!

15. Feeding studies conducted by academics:commercialised crops • Rats fed GM Bt corn over three generations: areas of necrosis to liver and kidneys and alterations in blood biochemistry (Kilic & Akay, 2008). • Old and young mice fed GM Bt corn MON810: marked disturbance in immune system cells and in biochemical (cytokine) activity (Finamore et al., 2008). • Pigs fed GM Bt corn variety MON810 for 31 days: differences in immune cell type numbers (e.g. CD4+ T cells, B cells, macrophages) and biochemistry (cytokine levels; e.g. IL-12, IFNg, IL-6, IL-4, IL-8) (Walsh et al., 2011). • Ewes and their lambs fed GM Bt corn variety Bt176 over three generations: hyperplasia of ruminal epithelial basal cells in ewes and a disturbed gene functioning of liver and pancreas in lambs (Trabalza-Marinucci et al., 2008).

16. Feeding studies conducted by academics:commercialised crops • Rabbits fed GM soy: enzyme function disturbances in kidney and heart (Tudisco et al., 2006). • Mice fed GM soy: disturbed liver, pancreas and testes function; abnormally formed cell nuclei and nucleoli in liver cells, indicating increased metabolism and potentially altered patterns of gene expression (Malatesta et al., 2002; Malatesta et al., 2003; Vecchio et al., 2004). • Mice fed GM soy over their lifetime (24 months): more acute signs of ageing in the liver (Malatesta et al., 2008); significant changes in the expression of 49 proteins; 39 proteins more expressed in GM-fed mice, 10 proteins decreased. Significant decrease in senescence markers (e.g. regucalcin, HSPs); lower metabolism. Structure of liver cell nuclei suggest marked lowering of gene function: GM Non-GM

17. Feeding studies conducted by academics:non-commercialised crops Mitogenic effect on gut mucosa of rats fed GM potatoes containing snowdrop GNA insecticide protein (Ewen SWB and Pusztai A, Lancet, 354, 1353-1354, 1999): GM Non-GM Rat Colon Rats fed GM Bt rice: significant differences in gut bacterial populations and organ weights (adrenals, testis, uterus) (Schrøder et al., 2007). “GM peas cause surprise allergic reaction”; unexpected post-translational modifications on bean a-amylase inhibitor in peas caused marked immune response and allergic type reactions in mice (Prescott VE et al. J Agri Food Chem., 53: 9023-9030, 2005).

18. Industry and academic led animal feeding studies show: Commercialised GM soy and corn show consistent signs of toxic effects in liver and kidney structure and function as well as some immune system disturbances. Such effects may be markers of the onset of chronic disease, requiring long-term rather than these reported short- and medium-term studies, to assess this more thoroughly. Unfortunately, such long-term feeding trials on GM foods are neither required nor requested by regulators anywhere in the world.

19. What could be causing these signs of toxicity? • 1. The introduced foreign GM gene (“transgene”): • GM gene product directly (e.g. Bt toxin) • Altered plant biochemistry caused by GM gene product (e.g. enzymes conferring herbicide tolerance) • 2. Higher exposures to herbicides used in conjunction with the cultivation of GM crops (e.g. glyphosate formulations) • 3. Altered plant biochemistry caused by mutagenic effect of the GM transformation process

20. Bt toxin • Crystalline protein complex (“Cry protein”) • Occurs naturally in the common soil bacterium Bacillus thuringiensis • Some types of Bt toxins are effective insecticides • Used as agricultural spray • Bt toxin in its native crystalline form is inactive as an insecticide; converted to insecticide active form in the digestive tract of certain insects • Bt toxin activation procedure: highly selective insecticide • Activated Bt toxin: inserts into and causes lesions in the insect’s gut epithelium; death either through a disrupted digestion or systemic bacterial infection

21. Native Bt toxin sprays vs. Bt toxin engineered into GM crops • GM Bt toxins: truncated active form • Bt toxin present throughout GM crop • GM Bt toxin approximately only 45% identical to the native form • GM Bt toxin in crops significantly different from that used as an • agricultural spray • Insect target specificity is compromised (e.g. see Schmidt et al., 2009) • Poses new health risks that need evaluating

22. Bt toxin insecticides in GM Crops:artificial and novel, coming from modified genes of a soil bacteria, a reservoir of more than 100 Bt toxin insecticides Ex. 44% difference in GM Bt176 maize

23. Why is Bt toxin a health concern? • Bt toxin: • Proven allergen and potent adjuvant in mammals (rats, sheep) even at low levels of exposure (Vázquezet al., 1999; Vázquez-Padrónet al., 1999 & 2000; Kroghsboet al., 2008; Adel-Patient et al., 2011; Trabalza-Marinucciet al., 2008). • Possesses properties, which with sufficient exposure could lead to allergic reactions caused directly by itself or against other ingested foodstuffs. • Immunogenic properties may account for the disturbing effects on immune system function observed in animal feeding studies (Finamoreet al., 2008; Walsh et al., 2011). • Bt toxin type Cry1Ab: • Present in commercialised GM crops (e.g. MON810 corn) • Binds to human cells in tissue culture • Disturbances in cell energy production and exterior (plasma) membrane systems leading to cell death, albeit at relatively high levels (Mesnageet al., 2012).

24. Human incidences of Bt toxin exposure • GM Cry9C Bt toxin “Starlink” corn, USA: • Intended only for animal feed; accidental entry into human food • Many recorded instances of allergic type reactions following consumption • Bt cotton, India: • Severe skin rashes in Bt cotton field workers; some cases needed hospitalisation (Gupta et al., 2005) • Farm animals feeding on the Bt cotton stubble suffering severe illness and death (Warangal District, Andhra Pradesh, 2006) • Bt toxin in pregnant and non-pregnant women, Canada: • Bt toxin found in the circulation of non-pregnant & pregnant women including blood supply to foetus • Source and integrity of the Bt toxin-unknown • Study shows that Bt toxin can survive digestion and enter the circulation. (Aris and Leblanc, 2011) • Consumption of Bt toxin GM food runs the risk of chronic systemic exposure. • Animal feeding studies suggest this may contribute to adverse health effects especially with respect to liver, kidney and immune system function. • Therefore, further investigation is needed before Bt crops can be claimed to be safe for humans.

25. Conclusions Increasing evidence shows the disruptive effect of the GM:need for in-depth molecular profiling to identify alterations in composition. Clear signs of toxicity: (especially to liver & kidney function) in controlled animal feeding studies even of a short-term nature. Cause of signs of toxicity: do they result from GM transgene function (Bt toxin, herbicide tolerance), herbicide residues, the mutagenic effect of the GM transformation process or combinations of these? Allergenicity: needs to be evaluated with human volunteers; no animal model systems available for this type of clinical investigation. Toxicity needs to be confirmed or refuted in life-long animal feeding studies. Based on available evidence and inadequacy of the tests requested by regulators, at present no GM crop and food can be categorically stated as safe to consume, especially on a long-term, life-long basis.

27. Background to Study • Follow up to industry 90 day feeding trial • Same experimental design: OECD, same strain & numbers of rats analysed • GLP conditions • Longer (2 years) • More parameters measured (e.g. endocrine hormone levels) • Three doses of GMO & Roundup • More extensive test groups; GMO, GMO+R, R • NOTE: toxicity NOT carcinogenicity study

28. Summary of Study Findings • Escalation of signs of liver and kidney toxicity in Monsanto 90-day feeding trial leading liver / kidney failure and premature death especially in males • Unexpected increase in tumor incidence, especially via Roundup in females (mammary tumors) • Unexpected low dose toxicity from Roundup (10,000 times lower than that permitted in drinking water in USA) • Females: died prematurely from mammary tumors (& pituitary dysfunction). • Note: severe adverse effects started at 4 months and peaked during second year of life.

29. Criticisms from GM Crop Advocates Wrong strain of rat: already cancer prone Too few rats analysed Wrong statistical methods of analysis Not in accordance with OECD guidelines But, Seralini study based on Monsanto study only longer and more comprehensive in analysis; either both are wrong or both are right as far as each goes!

30. Rebuttal of Criticisms For detailed, evidence-based rebuttal of criticisms: see gmoseralini.org

31. Conclusions • Re-evaluate assessment of NK603 GM corn: • Conduct carcinogenicity study with larger number of animals • Need to conduct life long (2-year) feeding studies for ALL GMO foods • Need to conduct life long (2 year) toxicity studies with complete pesticide formulations and not just active principle

32. The GM Transformation Process (A) PDS/1000 biolistic device used for microprojectile bombardment.(B) Suspension cells of tall fescue plated on filter paper before microprojectile bombardment.(C) Hygromycin resistant calli obtained after selection.(D), (E) Transgenic plantlets regenerated from the hygromycin resistant calli.(F) Transgenic tall fescue plants growing in the greenhouse.

33. Increased Rates of Mortality in GMO & Roundup Fed Groups

34. Increase Tumor Incidence in GMO & Roundup Fed Groups

35. Most Frequent Anatomical Pathologies

36. Increased Incidence of mammary Tumors in Females

37. Kidney Failure in Females

38. Kidney Leakage & Hormone Imbalance in Females Animals 33% GMO (solid line) vs Controls (dotted line): serum or urine measurements; at 15 month