1. RISK ASSESSMENT RESEARCH FOR THE PD PROJECT Thomas A. Miller 1, David J. Lampe 2, Ravi Durvasula 3 1 University of California, Riverside, CA 2 Duquesne University, Pittsburgh, PA 3 University of New Mexico, Albuquerque, NM Regulatory Communication Workshop 7-9 November 2006 UC Washington DC Center Thomas.miller@ucr.edu

2. My name is Tom Miller 1. Professor of Entomology. Teach Insect Physiology and Insect Toxicology. 2. Entomologist in the Agricultural Experiment Station. Do something about plant pests and disease. Thomas.miller@ucr.edu

3. Hurricanes in 2004 spread the citrus canker pathogen, Xanthomonas axonopodis from containment in the 2 southeastern counties of the state where a $500 million program had pushed it. August 25, 2005 Los Angeles Times “The only defense is containment through destruction.” “We had to destroy the village to save it” Attributed to many different people, including war correspondent Peter Arnett who supposedly attributed the quote to an unidentified Army officer. Used circa 1968, perhaps during the bloody Tet offensive. Thomas.miller@ucr.edu

4. "California is particularly vulnerable to non- indigenous pests and diseases," Daniel Sumner, a UC Davis agricultural economist and director of the Agricultural Issues Center. Thomas.miller@ucr.edu

5. A new pest comes into California every 60 days. Glassy-winged sharpshooter (GWSS) arrived ≈ 1980s? Pierce’s disease first noted in Anaheim in 1884 [“Orange County” disease]. Thomas.miller@ucr.edu

6. Total economic impact of more than $170 million a year, the Texas wine industry has more than 85 wineries and is the nation’s fifth-largest wine producing state." Pierce’s Disease The most serious threat to wine grapes in Texas, California and other growing areas is Pierce’s disease, which occurs in all areas of Texas that do not have severe winters. Pierce’s disease, which can wipe out a vineyard. Currently, there is no preventative or curative treatment for it. Efforts to reduce the risk of Pierce’s disease include controlling the glassy-winged sharpshooter. Site selection and chemical control help prevent the pathogen’s spread. Although a large research effort is underway to discover methods for controlling Pierce’s disease, it will probably be a long time, perhaps 10 years or more by some estimates, before a practical, effective solution is available. Florida grape varieties are resistant to many diseases, principally Pierce's disease (PD), a disease that devastates Vitis vinifera. Soon molecular genetic research will produce further improved varieties and possibly enable traditional wine grape production in our state. Bryon Biddle Three Oaks Winery, Vernon, FL 20 October 2006 Thomas.miller@ucr.edu

7. Pierce’s disease control program annual report to the legislature 2005 Thomas.miller@ucr.edu

8. Grape acreage by County in California 2005 “back in 1999-2000, people … were saying that the grape growing industry was dead in Temecula and there … will be a 100% loss of vines due to PD.” -- Nick Toscano Thomas.miller@ucr.edu

9. Nick Toscano Specialist & Extension Entomologist email: nick.toscano@ucr.edu (951)827-5826 (951)827-3725 nick.toscano@ucr.edu “ … Callaway [vineyards] lost about 100% … close proximity to 120 acres of grapefruit … “800 out of a total of 2500 acres of vines were removed from Temecula in 2001. Since I initiated and directed the program, 300 new acres have been planted. The number of wineries have increased from 13 to 21 (60% increase). -- Nick Toscano 12 October 2006 Thomas.miller@ucr.edu FISCAL YEAR 2003/04 2004/05 2005/06 REVENUE State (Budget Act) 6,408,000 4,408,000 4,341,000 Federal (USDA) 9,885,525 14,903,606 15,300,000 Board Assessment 968,000 305,000 343,400 Total Resources 17,261,525 19,616,606 19,984,400 EXPENDITURES Personal Services 3,053,409 3,456,754 3,658,520 Operating Expenses 2,206,328 3,040,503 2,852,186 County Payments 12,001,788 13,119,349 13,473,694 Total Expenditures 17,261,525 19,616,606 19,984,400

10. 1. Find a symbiont (blue) cycling with the pathogen (orange). 2. Design toxins against Xylella. 3. Disease cycle testing. Endophyte Candidate: Alcaligenes xylosoxidans Tim Yolo and Arinder Arora, UC Riverside Symbiotic control of PD Dave Lampe Duquesne University, Pittsburgh PA 2001-2005 Carol Lauzon CSUEB Hayward CA DsRed Alcaligenes (RAxd) Axd S1 RAxd 2004 (+) 2001 Thomas.miller@ucr.edu 2003: Field testing of RAxd?

11. Where is risk assessment done? 1.In the laboratory at BL-1. 2.In the field. Laboratory data poorly predict field results. 1.GWSS die in 3 weeks in the lab. 2.Grapevines do poorly in greenhouses. 3.It may be impossible to simulate citrus next to vineyards.

12. We want to test behavior of RAxd in grapevines (2003). Okay, but you have to burn the grapevines at the end of the season.- EPA Field testing for Risk Assessment Then how will we know if the disease is cured or not? We don’t make the laws. (National Environmental Protection Act 1969; FQPA 1996; PPA 1990; TSCA 1976; FIFRA 1972). The UCR Biosafety Committee approved of working with RAxd at BL-1 level. Why are we burning grapevines? “… appropriate for working with microorganisms that are not known to cause disease in healthy human … municipal water-testing laboratories, in high schools, and in some community colleges teaching introductory microbiology classes …” Thomas.miller@ucr.edu

13. "The Biosafety level is chosen to be commensurate with the potential risk. Rather than ensure absolute containment of every experiment, which would be onerous for researchers, containment safeguards are allowed to be more relaxed when the risk is small. It is recognized that BL1 may not ensure containment all of the time, but that is acceptable since (for example) damage caused by release is considered insignificant and the likelihood of the organism to survive outside the lab is also considered low." -- Howard Judelson Professor of Plant Pathology UC Riverside 23 October 2006 BioSafety ruling incompatible with regulation? Thomas.miller@ucr.edu

14. Needle inoculation of grapevines, Napa, CA summer 2003. Bagged grapevines, Temecula, CA 2004 Thomas.miller@ucr.edu

15. Movement of Alcaligenes in Host Plants Plant type Detected/tested: amount* found Lemon 25/25: 3,591,427 cells/2cm Orange 25/25: 943,305 cells/2cm Crepe Myrtle 8/25: 884,770 cells/2cm Periwinkle 10/25: 304,820 cells/2cm Grapevine 24/25: 18,225 cells/2cm Movement three inches away in two weeks; Lemon preferred * Cells in 2 cm of plant stem Thomas.miller@ucr.edu

16. The GloFish Los Angeles Times, front page, 22 November 2003 Normal Zebra fish Thomas.miller@ucr.edu

17. Can we sell DsRed zebra fish as pets? -- 2003 Let’s see, okay go ahead, we can’t stop you, there is no law against that. — USDA, FDA, EPA, CDC, NIH Regulatory wars Hey, that’s the same transgene as in RAxd. Why don’t you make them burn the fish tanks? We don’t make the laws. Thomas.miller@ucr.edu

18. November 2003 Thomas.miller@ucr.edu

19. http://ucbiotech.org/ Thomas.miller@ucr.edu Regulatory agencies don’t need to worry about the “values” question; others are already doing that.

20. Environmental Impact of Transgenic Alcaligenes Used to Control Pierce’s Disease. Miller, UCR; Durvasula, Yale; Lampe, Duquesne September 2004. Identification of hazards. Fitness alteration Transfer to non-target organisms: Transgene instability Assessment of community ecology impacts. 1. Creating transgenic Axd strains. 1a. Attenuated strains. 1b. Cysteine auxotrophic strains 2. Measuring the rate of horizontal gene transfer 3. Monitoring for HGT in natural environments by RT PCR. 4.Reducing the likelihood of HGT; genetic toxin- antidote. Testing of ecological impact of transgenic Axd. 5.1. Bacterial census of grapevine. 5.2. Changes in bacterial census (grapevine). 5.3. Bacterial census of grape rhizosphere. 5.4. Changes in bacterial census (rhizosphere). 6. Modeling transgenic Axd in the environment. Amount requested: $200,000/yr Amount funded: $100,000/yr Term: 2 years. Study what researchers think is important. Thomas.miller@ucr.edu

21. 4 events in 2004 Pew Foundation 22 Jan 2004 Business Week 3 May 2004 NAS Dec 2004 “ … likelihood of success [of symbiotic control] is limited …” Regulation of transgenic insects is unprepared. NAS Dec 2004 “… science knows too little about how the novel organisms will behave in the real world.” Regulatory and public discourse centers around the “unnaturalness” of genetic transformation. Thomas.miller@ucr.edu

22. COMMITTEE ON CALIFORNIA AGRICULTURAL RESEARCH PRIORITIES: PIERCE’S DISEASE ___________________________________________________________ JAN E. LEACH, Chair, Kansas State University, Manhattan PEDRO BARBOSA, University of Maryland, College Park MICHAEL J. DAVIS, University of Florida, Homestead DAVID G. HOEL, Medical University of South Carolina, Charleston L. JOE MOFFITT, University of Massachusetts, Amherst ALISON G. POWER, Cornell University, Ithaca TERRY L. ROOT, Stanford University, Stanford JACK SCHULTZ, The Pennsylvania State University, University Park WILLIAM F. SPLINTER, University of Nebraska, Lincoln BRIAN J. STASKAWICZ, University of California, Berkeley MARIE-ANNE VAN SLUYS, University of São Paolo, Brazil T. ULF WESTBLOM, Central Texas Veterans Health Care System, Texas A&M University, Temple Staff KIM WADDELL, Study Director (through March 2004) ROBIN SCHOEN, Senior Program Officer MICHAEL KISIELEWSKI, Research Associate DONNA WILKINSON, Research Intern PETER ROGERS, Research Intern CINDY LOCHHEAD, Project Assistant JULIE COFFIN, Project Assistant Pages 109-110: “Using paratransgenesis to manage PD clearly would be along-term strategy, and one in which the likelihood of success is limited. Although some progress toward transformation of GWSS endosymbionts has been made (Lampe and Miller, 2002), the committee views this as Category 4 research, in addition to its scientific uncertainty, there are ecological and regulatory barriers to success that are at least as significant as any technical barriers.” This is exactly what “they” said about transgenic pink bollworm. NAS Dec 2004 Thomas.miller@ucr.edu

23. Frances Oldham Kelsey, Ph.D., M.D., (b. 24 June 1914) is a naturalized American pharmacologist, most famous as the reviewer for the U.S. Food and Drug Administration (FDA) who refused to authorize thalidomide for market because she had concerns about the drug's safety. Her concerns proved to be justified when it was proven that thalidomide caused birth defects. Kelsey's career intersected with the passage of laws strengthening the FDA's oversight of pharmaceuticals.24 June 1914naturalizedAmericanpharmacologistFood and Drug Administration thalidomide birth defects pharmaceuticals Frances Kathleen Oldham Kelsey receiving the President's Award for Distinguished Federal Civilian Service from President John F. Kennedy, in 1962 John F. Kennedy Thomas.miller@ucr.edu

26. 13. Regulatory Issues with Biotechnology (Thomas A. Miller) 1) Margaret McFall-Ngai, Univ. Wisconsin, Symbiosis as an emerging field [invited, no reply] 2) Angela Douglas Univ.York, UK, Symbiosis as an emerging field [accepted in another session] 3) David Brooks, Oxitec, UK Transgenic insects [invited] 4) FDA and tooth decay, Jeffrey Hillman, Oragenics, FL, USA [accepted, health allowing] 5) Shrimp aquaculture, Ravi Durvasula, Univ. of New Mexico [accepted] 6) Chagas disease, Ravi Durvasula, Univ. of New Mexico. [accepted] 7) India Regulatory process, lead by K. P. Gopinathan, Bangalore, India [invited] 8) China Regulatory process, lead by Yongping Huang [accepted] 9) Thailand and Laos Regulatory process, Yupa Hanboonsong [accepted] 10) USA Regulatory process, Janet L. Andersen, EPA-BPPD [invitation pending] 11) USA Regulatory process, Robyn Rose, APHIS-BRS [invitation pending] 12) British Regulatory process, Angela Douglas [accepted] 13) Canadian Regulatory process, Brian Belliveau, Health Canada [recruiting someone else] 14) Greek Regulatory process, Hanneke Drosopoulos [accepted] 15) Korean Regulatory process, Ministry of Environment [accepted – being organized] 16) French Regulatory process, [asked Rene Feyereisen to suggest someone] 17) Morocco Regulatory process, Malika Bounfour [invitation pending] 18) Slovenia Regulatory process, Prof. Dr. Maja Ravnikar [accepted]