INTERNATIONAL SEED TESTING ASSOCIATION ISTA www seedtest org

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org The International Seed Testing Association GMO PT Program AIEC Fall Meeting October 15 -16 2014 New Orleans, Louisana

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org What is the International Seed Testing Association? • An international, non-profit association of persons and seed testing laboratories from the public and the private sector • Under the Governance of the Governments of member countries and distinct economies • An Association standardizing the determination of seed quality to facilitate international trade

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Ordinary Meeting Voting delegates from 72 Member Governments Executive Committee 11 Members from all Continents of the World nominated by Governments Secretariat 11 professional employees 18 Technical Committees and Boards Over 200 experts from all over the world working voluntarily Experts from the public/private sectors, from member/non-member laboratories

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org ISTA Executive Committee 2013 -2016: President Vice President 9 Members-at-Large Joël Léchappé, France Craig Mc. Gill, New Zeeland Mable Simwanza, Zambia Steve Jones, Canada Alexander Malko, Russia Berta Killerman, Germany Masatoshi Sato, Japan Grethe Tarp, Denmark Margus Friedenthal, Estonia Rita Zecchinelli, Italy Cecilia Jones, Uruguay

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org ISTA Secretariat Name: Role: Benjamin Kaufman Secretary General Jonathan Taylor Nadine Ettel Publication Specialist TCOM Coordinator Rasha El-Khadem Head of Accreditation Dept. Branislava Opra Florina Palada System Auditor Patricia Muschick Head of Administration Dept. Cannice Gubser Agnes Hegedüs Sejal Patel Membership Administration Sales- and Office Administrator Olga Stockli

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org ISTA Technical Committees: • • • • • ADVANCED TECHNOLOGIES COMMITTEE BULKING AND SAMPLING COMMITTEE FLOWER SEED COMMITTEE FOREST TREE AND SHRUB SEED COMMITTEE GERMINATION COMMITTEE GMO COMMITTEE MOISTURE COMMITTEE NOMENCLATURE COMMITTEE PROFICIENCY TEST COMMITTEE PURITY COMMITTEE RULES COMMITTEE SEED HEALTH COMMITTEE STATISTICS COMMITTEE SEED STORAGE COMMITTEE TETRAZOLIUM COMMITTEE VARIETY COMMITTEE VIGOUR COMMITTEE EDITORIAL BOARD (SST)

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Why is ISTA relevant to seed stakeholders? The vision of ISTA: Uniformity in Seed Quality Evaluation Worldwide

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International standardization of reporting results International exchange of scientific results International standardization of methodologies International standardization of laboratory performance

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International exchange of scientific results: ISTA Publications and Symposia • Editing and issuing the Scientific Journal: Seed Science and Technology • Organize Seed Symposium as part of the tri-annual ISTA Seed Congress

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International standardization of reporting results International exchange of scientific results International standardization of methodologies International standardization of laboratory performance

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org 201 3 : Ne of Methodologies: International Standardization w t o The International Seed Testing C thfor Cha Rules h a e -> ISTA has been publishing the international rules for pte ISTA pter r 19 a seed testing since 1933. d R u -GM les! ded OT ! esti ng

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Chapter 19: GMO Testing • Objective: – To formalize the ISTA approach for GMO testing within the ISTA Rules – To describe a system that will ensure uniformity in GM seed testing • Why develop a new chapter? – – ISTA has been working to address challenges related to testing GM seed since 2001! GMO testing is an increasing activity for seed trade Laboratories need guidance about ISTA‘s principles and technical aspects of GMO testing. Answer ISTA‘s strategy decision (2001: to establish a GMO chapter) • Key Considerations: - This chapter remains general on all aspects of GMO seed testing This chapter is based on already existing ISTA‘s papers Doesn‘t provide protocols (follows the Performance Based Approach) Is linked to a Handbook for the detailed technical considerations (in development!) Is not creating new ISTA position / statement about GMO analysis

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International standardization of reporting results International exchange of scientific results International standardization of methodologies International standardization of laboratory performance

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International Standardization of Laboratory Performance: The ISTA Accreditation Program: • Open to : Seed Testing Laboratories from the public and the private sector • official language: English • all relevant documents can be downloaded from the ISTA website at www. seedtest. org

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International standardization of reporting results International exchange of scientific results International standardization of methodologies International standardization of laboratory performance

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International Standardization of Reporting Results: Facilitation of International Seed Trade • ISTA Certificates have international recognition • Indicate the sampling and testing has been carried out in accordance with the ISTA Rules • Sampling and testing methods used have been validated, internationally harmonised and voted on by the ISTA membership.

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org International standardization of reporting results International exchange of scientific results International standardization of methodologies International standardization of laboratory performance

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org The ISTA Accreditation Programme ISTA Membership ISTA Rules ISTA Certificate ISTA Accreditation Programme Q-Audit Proficiency Testing § Quality assurance system according to the ISTA Accreditation Standard § ISTA audit every three years § Audit by two ISTA auditors § 120 participating laboratories world wide § The laboratory‘s performance is directly linked to its accreditation status § Three test rounds per year Seed Testing Laboratory

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org The ISTA Proficiency Test Program Basic Tests Seed Health Tests GMO Tests

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org The ISTA Proficiency Test Program Goal: measure and monitor laboratory‘s proficiency • Proficiency Test fee is covered in yearly Membership fee • Participation is mandatory for accredited labs, for the tests/crop group/events for which they have been accredited • Voluntary participation possible for all member laboratories, regardless if accredited or not • Non members are welcome to participate (service fee will be charged)

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org The ISTA Proficiency Test Program: • Several rounds per year • Labs test ‘blind’ samples in accordance with determined methods* • Submission of results • Review and Statistical evaluation • Evaluation of the laboratory‘s proficiency • Detection of systematic errors and performance trends • Results influence accreditation status (only labs for which participation is mandatory) Accreditation maintained? þ

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org The ISTA Proficiency Test Program: Ratings • Ratings are assigned: A, B, C and BMP (Below Minimum Performance) for each test in each round • After six rounds of participation calculation of overall rating • Only mandatory test results are included in overall rating • Performance of a laboratory influences their accreditation: Overall rating of C warning letter Overall rating of BMP suspension Failure to participate in 2 rounds out of 6 suspension

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO Proficiency Testing: Challenges! • ISTA’s performance based approach for GMO testing means the way development and execution of a PT used for Basic Seed Tests and Seed Health Tests must be modified in order to have a Proficiency Testing program for GMO testing

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO TESTING: Performance Based Approach • The ISTA approach has targeted the uniformity in GMO testing results, not by the uniformity in testing methodology, by using a “Performance Based Approach (PBA)”. • The “Performance Based Approach” requires that laboratories demonstrate that the GMO detection, identification or quantification methods they are using on seed samples for reporting results on the ISTA International Seed Lot Certificate, meets acceptable standards set by ISTA. • This standard includes, among others, sampling, reporting and test accuracy. 24

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org What does this mean? • Basic Seed Tests and Seed Health Tests must use ISTA approved protocols… but…. • …the Performance Based Approach gives seed testing laboratories the choice to use different technological approaches, e. g. bioassays, protein based methods and DNA based methods. • The GMOPT must be able to accommodate evaluation and rating of results obtained from many test types!

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO PT Program basics: • 1 species per round, 1 -3 Events • Qualitative and Quantitative evaluations • Spiking levels between 0 -4% • 10 -12 samples of 1800 -3000 seeds each • Two panels per year offered • Matrix is SEED

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO PT 19: Design of Round

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Announcement and Technical Protocol: Announcement is made on the ISTA Website and also sent to member labs via e. mail Technical protocol provides information needed to develop the test plan http: //seedtest. org/en/proficiency-tests-_content---1 --1157. html Reporting sheets are provided for results to be returned

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org ISTA GMO PT Database The PT process is tracked and executed through the use of a specially developed Access database which is used for registration to sample prep, shipping, participant tracking over time, data recording and analysis, etc.

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Results Evaluation: Qualitative Determination • The rating for the presence/absence ( yes/no - qualitative) results is based on a % of misclassified samples/total samples. • Misclassification may either be a false positive or a false negative result. • Missing results for individual samples are considered as misclassification.

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Results Evaluation: Quantitative Determination • The quantitative rating is based on the quantification results for the positive samples and their reference value (samples with zero spiking level are not used in quantification rating). • The reference value is either the # of GM seeds in %, the mass of the GM seeds in % or the median of the results reported by the participants in the unit ‘%DNA copies’. • Which of the reference values is chosen is determined by the panel of experts appointed for each round, the guiding principle being: • Sub-sampling quantification: %number • Results reported in %number: %number • Results reported in %mass: %mass • Results reported in any other unit, such as number DNA copies: median of the results

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org ISTA GMO Proficiency Test – Ratings The rating system is based on the system established for basic and seed health tests which are accredited by ISTA – A, B, C, BMP A B C BMP No problem has been detected in this test There are small problems, but no specific look or action is suggested to the participant Problems, ISTA indicates there might be things to consider by the laboratory to explain or correct things Below Minimum of Performance, ISTA indicates that the results were poor and the laboratory need to find explanations and to improve/correct

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO PT 19: Results/Evaluation Sample sets were sent to 46 laboratories; 39 participants submitted results; 8 provided qualitative results only PT 19 - Ratings Quantitative Results 40 20 30 15 # of Labs PT 19 - Ratings Qualitative Results 20 10 10 5 0 0 A B C BMP

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO PT: Qualitative and Quantitative Rating Systems STI 130: -2005: https: //www. seedtest. org/upload/cms/user/STI 130 Oct 2005. pdf STI 128 – 2004: https: //www. seedtest. org/upload/cms/user/STI 128 Oct 2004. pdf Pro ced upd ure ated

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Success of the Performance Based Approach for GMO Testing within ISTA: 13 laboratories are accredited for GMO Testing Performance Trends: Qualitative Detection A B C BMP 35 30 25 20 15 10 5 0 PT 11 PT 12 PT 13 PT 14 PT 15 PT 16 PT 10 PT 11 PT 12 PT 13 PT 14 PT 15 PT 16 60 50 40 30 20 10 0 Quantification A B C BMP

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO PT Program: Participant Stats: About half the participants are ISTA labs The majority of participants are from Europe

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Complete process: 1 round of the GMO PT program: The execution of a single round of the GMO PT Program is complex and involves significant organization, coordination, and administration from all parties involved in it’s delivery. Steps include: 1. Program Planning and Seed procurement 2. Purity testing of sample seed material 3. Announcement of PT Round 4. Initiation of a new PT in the PTDB 5. Registration of participants and invoicing 6. Sample preparation and shipment 7. Participant Results 8. Evaluation of Results and Ratings 9. Generation of Final results reports 10. Record keeping Adding to the complexity – there always 3 PT rounds ongoing at any time, all at different stages of completion!

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org How is this work accomplished? Working Lead Members Groups GMO ISTA Christoph Proficiency TCORD Haldemann, Test: Nadine Jean-Louis Administration Ettel Laffont and Sample Cheryl Dollard Preparation (Expert Team); Andrea Jonitz (Sample Prep) GMO Ray Elizabeth Proficiency Shilito Bates (Bayer), Test: Bruno Resource Zaccomer Procurement (Monsanto), Sofia Ben Tahar (Limagrain), Work Description and Projects Purpose: Execution of the GMO PT ; Lead: Coordinate administration of the PT program from within the ISTA secretariat; Expert Team: Develop PT Plans, Evaluate Results and Ratings, address technical questions, assist with administration of program as described in the SOP for administration of the GMO PT Program. Sample prep: Purity check of material and preparation of PT Samples as described in the GMO PT Program Sample Prep SOP Purpose: Resource procurement in support of the GMO PT Program: Secure reference materials (GM and non-GM) from Industry; Coordinate financial support for PT program from Industry/corporate partners; Action items: solicit WG members from other Industry areas: Syngenta, Pioneer, etc Secretariat contact: ISTA TCOM Coordinator (Nadine Ettel)

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org GMO PT Program: Procedures established Two SOPS developed describing all steps required to execute a single PT round: “Administration of GMO PT program” “Sample preparation for GMO PT program” Significant challenges: Sourcing GM and Non-GM Seed; Funding to cover associated costs Finding labs willing to prepare the samples

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org One Round of the GMO PT. . Mostly! K. Studer

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Sample Preparation • A sample preparation laboratory prepares the blind samples, packages and ships lots to participants. • Requires Seed Lot Purity Checks: GM and Non-GM Seed • Preparation and blind coding of the spiked samples (10 samples x 50 participants – 500) • Arrangement of phytos • Rounds 1 -19: LTZ (Germany), Round 20: Eurofins

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Expert Team • Group of experts who are involved in statistical analysis and interpretation of the results for each round • They address the activities which cannot involve other committee or WG members in order to ensure secrecy of the program, and to protect confidentiality of participants and results. • Includes: finalizing details and logistics to addressing questions and issues that arise and are unique in each PT.

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Expert Team: A snapshot… • • • Development of PT test plans (computing percentages/spiking levels/seed requirements/statistical verification). Provision of support for Administrative functions carried out in the Secretariat as required (preparation/review of documents/files, and database use, generation of reports, etc). Preparation of the PT schedule Coordination of activities and problem solving issues related to seed procurement, including addressing Industry questions regarding seed used for the PT program. Verification of seed check procedure; analyse data for approval of lots; revision of test plans when necessary. Provided input for development of new GMODB; beta tested versions during development process. Review of raw results, ratings computation and determination of final ratings. Provision of advice and answers to the Secretariat, Sample prep lab and members with questions. Review of data for trends that signal potential issues which could require modification of the Program. ** “Corporate Memory” ** Addressing issues related to the PT: These are often complex and require technical analysis, research, etc.

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org 20 rounds and counting… PT Crop # of Events # of participants PT 01 PT 02 PT 03 PT 04 PT 05 PT 06 PT 07 PT 08 PT 09 PT 10 PT 11 PT 12 PT 13 PT 14 PT 15 PT 16 PT 17 PT 18 PT 19 PT 20 Maize Soybean Canola Maize Soybean Maize* Cotton** Canola Maize Canola Soybean Flax**** Maize***** Soybean Maize+ Soybean+ Cotton++ 2 1 2 1 2 2+1 2 3 2 1 1 1 2 2 46 50 42 52 58 50 67 53 64 30 49 60 31 40 35 53 45 48 46 in progress # of samples in a set 30 10 12 12 12 10 12 14 12+2 12 12 12 8 9 8 8 8 in progress # seeds per sample 3000 1500 3000 3300 2000 3000 2000/50 2500 3000 2000 3000 1500 1800 2000 2500 1800 in progress # spiking levels (incl. 0) 4 3 7 4 5 4 6 5 11 9 6 8 7 6 4 4 5 4 4 in progress

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Ordinary Meeting Voting delegates from 72 Member Governments Executive Committee 11 Members from all Continents of the World nominated by Governments Secretariat 11 professional employees 18 Technical Committees and Boards Over 200 experts from all over the world working voluntarily in the Technical Committees Experts from the public/private sectors, from member/non-member laboratories

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Conclusion • ISTA GMO PT program is a unique tool for achieving uniform and reliable seed quality determination for GMO testing • It is possible to evaluate and compare test performance internationally in the dynamic field of GM testing internationally while still achieving uniformity of results reporting that is ISTA’s aim.

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Thank you for your attention www. seedtest. org

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) www. seedtest. org Questions ?