1. Animal Wholesomeness studies Wilna Jansen van Rijssen PhD GMASSURE 23 - 25 November 2015

2. Purpose of studies Animal studies to evaluate input trait from GM crops Animal studies to evaluate output traits from GM crops

3. To confirm the nutritional value of the GM crop

4. USE OF ANIMAL FEEDING STUDIES: GROWTH PERFORMANCE Detection of potentially unintended effects Broiler studies are standard component of assessment – safety and nutrition Rapid growth period when sensitive to SMALL nutritional or anti-nutritional changes

5. INPUT TRAITS Crop protection against diseases, insect damage, tolerance to herbicides OUTPUT TRAITS Nutritionally enhanced Reduced levels of toxicants

7. POULTRY MEAT PRODUCTION

8. FLOW CHART: GM diet preparations SEED Planting/ growing/h arvest/ storage Analysis Process Analysis Formulate diets Manufac turing diets Animal/ product assessmen t Data analysis Report /sampling retention LIMITING VARIABILITY - STATISTICAL ANALYSIS - NUMBER OF ANIMALS

9. POULTRY: Diet Starter diet Grower diet Finisher diet

10. Production, Handling, Storage, Processing crop Sampling, analysis Statistical analysis and interpretation of results

11. QUALITY CONTROL: Production, Handling, Storage, Processing crop  SOPs / records  Planting (planting design, maintenance of plots)  Field plots  Growing season  Agronomic performance  Harvesting  Transport equipment  Grain storage locations  Unloading  Storage (e.g. storage moisture)  Sample  Crops processing

12. QUALITY CONTROL: Sampling (internationally agreed methods) Sample lot Grain sampling (probe) Hay sampling Green chopped forage sampling Pasture sampling Mixed diet sampling Handling of samples

13. QUALITY CONTROL: Analysis Analysis of : ▫Introduced trait ▫Pesticides, mycotoxins ▫Nutrients, anti-nutrients ▫Toxicants Analytical methods: ▫Chemical, microbiological

14. NUTRIENT ANALYSIS Crops/grain/ Co-products Livestock typeAnalyte Grain: maize, wheat, barleyNon-ruminantsxxxxxxxxxxxxxx Oilseed meals, soybean, linseed, cottonseed, canola meal, full-fat oilseeds Non-ruminantsxxxxxxxxxxxxxx Grain: maize, wheat, barleyRuminantsxxxxxxxxxxxx etc

15. Anti-nutrients in crops CROP OF PRODUCTANTiNUTRIENT Soy bean / soy bean mealTrypsin inhibitors / lectins Canola / canola meal Rapeseed / rapeseed meal Glucosinolates Cotton, cottonseed, cottonseed mealGossypol, cyclopropenoid fatty acids

16. MYCOTOXINS Aflatoxin B1Fusarenon X Aflatoxin B2Deoxynivalenol (DON) Aflatoxin G115- acetyl - DON Aflatoxin G23-acetyl-DON Ochratoxin ANivalenol CitrininZearalenone T-2 toxinFumonisin B1 HT2-toxinFumonisin B2 DiacetoxiscirpenolFumonisin B3 Neosolaniol

17. ACCREDITED LABORATORIES GOOD LABORATORY PRACTICES (GLP) ▫OECD ▫SANAS METHODS AOAC official methods e.g.

18. STATEMENTS: signed Study leader Members of team Forms – transfer (chain of custody), storage, field characteristics, plot plan, field history fertilizers, herbicides

19. STATISTICAL ANALYSIS Hypothesis – basic objectives, what to accomplish Treatments Experimental units (pens?) Measurements Experimental design Can the experimental design be analysed properly?

20. ESTIMATED NUMBER OF REPLICATIONS (BLOCKS) NEEDED TO DETECT A TREATMENT DIFFERENCE AT P<0.05 Coefficient of variation (%) EXPECTED DIFFERENCE (%) 510152025 2 432-- 37332- 4124332 5176433 6247433.. 30566142633724 Randomized complete block design with 2 treatments, two-tailed test of significance at P<0.05, 80% power

21. PERFORMANCE, CARCASS YIELD, MEAT QUALITY ItemGM soybeanControlReferences (6) PERFORMANCE Live weight D0 Live weight D42 Feed intake (kg/bird) & kg/pen) Feed conversion factor (kg/kg) CARCASS YIELD (relative) Live weight/fat/thigh/ drum/wing etc QUALITY Moisture/protein/fat (breast/thigh

22. INTERPRETATION Non-statistical knowledge should be incorporated Design and analysis should be kept simple Difference between statistical and practical significance is important

23. Interpretation of experimental results ▫Researchers with background and training to interpret results ▫Historical data – same lab and other labs

24. Increased nutrient concentrations Decreased toxicant concentrations

25. Case study Comparison of performance and carcass parameter when fed diets containing soybean meal produced from GM-soy bean (HIGH OLEIC ACID) control (nearest isoline) or conventional reference soybeans (6 lines)

26. Poultry meat production Nutrient requirements Understanding of nutrient digestion, metabolism, intermediary metabolism, nutritional physiology, pathology and nutrient flow and retention in the bird Management of broiler performance ▫Climate conditions, water quality

27. Poultry meat production Diet formulation and processing Particle size, enzyme supplements, change in starch matrix (e.g. effects on gelatinization of the starch), anti-nutritional factors, including growth promoters etc Digestibility studies (increased amounts of cellulose or hemicellulose, increase amount of amino acids Bioavailability studies (digestion, absorption, metabolism Sensory (organoleptic) evaluation

28. PERFORMANCE /CARCASS PARAMETERS GM soybean meal (increased level of oleic acid) compared with control and references Different levels of GM soybean ? Parameters Weight gain – feed intake – feed conversion Organs and muscle weights as % of carcass weight Fatty acid composition of animal fat Liver enzymes (GOT etc) liver damage etc

29. CONCLUSION The results of this study support the conclusion that there are no differences between the GM meal and the control and the conventional references. Therefore the GM meal is equivalent to conventional in the marketplace. This study confirms the results from extensive compositional analysis which showed no nutrient and antinutrient differences between the GM and control. The data support the conclusion that GM crop is as wholesome and nutritious as the conventional crop on its ability to support rapid growth and equivalent carcass yield

30. International harmonisation of study protocols for livestock and poultry performance ILSI Best Practices ….animal studies……GM crops…output/input traits, 2007 VICH (2000) Good clinical practices, VICH GL9 (GCP) Available http://vich.eudra.org/pdf/2000/G109_st7.pdf http://vich.eudra.org/pdf/2000/G109_st7.pdf Nutrient requirement of domestic animals - cattle, chickens, etc, National Academy of Science, USA Taylor et al (2007) Broiler performance and carcass parameters – diets containing GM soymeal, Poultry Science, 86:26-14