Document D6pm411Y5V3pEdDKYqbDv1dd

To: Beck, Nancy[Beck.Nancy@epa.gov]; Pruitt, Scott[Pruitt.Scott@epa.gov]; Brown, Byron[brown.byron@epa.gov]; Jackson, RyanOackson.ryan@epa.gov]; Dravis, Samantha[dravis.samantha@e pa.gov] Cc: Gay Ludwick[gay@gdcillc.com]; Roy Bailey[rbailey@gdcille.com] From: Roy Bailey Sent: Sun 7/16/2017 3:36:39 AM Subject: Attention please EPA Improperly Granted the Wolbachia EUP.DOCX 0001.htm Administrator, I hope you are well. I want to bring to your team's immediate attention something that hopefully you and your team will appreciate. I have included an attachment for your review. The timing is now and hopefully after reviewing the attachment you will consider canceling the field trials until the potential human and environmental impact of the mentioned technology can be reviewed and studied. There are some very smart and experienced people who understand this technology backwards and forwards and they believe, once reviewed, it won't pass muster. At a minimum, a pause and some time to study it could avoid a disaster and embarrassment to your agency. I will be more than pleased to put our friends from Intrexon on a call immediately to discuss. After all, they have been in this mosquito business for over 10 years and have extensive knowledge of the subject matter. Thank you and your team for considering this request and for your attention to the concerns outlined in the memo. All my best regards and respect Roy W. Bailey CEO Giuliani Deason Capital Interests, LLC Cell 214.208.1721 Office 214.378.3663 Rbailey@gdeillc.com Rbailev@bailevstrategicadvisors.com Begin forwarded message: From: "Bobo, Jack" <J6obo@mtrexon.com> Date: July 15, 2017 at 6:59:38 PM PDT 17cv1906 Sierra Club v. EPA - 6/22 Production ED 001523 00003112-00001 To: 'Roy Bailey' <rbailey@,gdcille.com> Subject: Memo CONFIDENTIAL TRANSMISSION - To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by Intrexon Corporation to be confidential and proprietary. This communication is expected to be read and/or used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. 17cv1906 Sierra Club v. EPA - 6/22 Production ED 001523 00003112-00002 EPA Must Revoke the Improperly Granted Wolbachia Aedes aegypti Experimental Use Permit EPA's Office of Pesticide Programs improperly granted an Experimental Use Permit (EUP) to MosquitoMate allowing releases of millions of Wolbachia-infcctcd Aedes aegypti mosquitoes in California and Florida [1], Oxitec and other commenters submitted substantive comments on the record identifying significant concerns and critical risks associated with the proposed EUP. Notwithstanding these comments, EPA granted the requested EUP to MosquitoMate. EPA can immediately remedy this improper regulatory approval and all other current Wolbachia EUPs. Pursuant to FIFRA Section 5(e) (7 U.S.C. 136c(e)), the EPA Administrator may revoke an EUP immediately if he finds that its terms or conditions are being violated or that its terms or conditions are inadequate to avoid unreasonable adverse effects on the environment. (See also 40 C.F.R. 172.10(a)) EPA's regulations provide that a permittee may "contest" the notice of revocation by requesting "an opportunity to confer with the Administrator." The Administrator will issue a final decision within 20 days after such conference. (40 C.F.R. 172.10(c)) Neither FIFRA nor EPA's regulations provide for a formal hearing to object to revocation of an EUP. EPA's granting of an experimental use permit to MosquitoMate for an Aedes aegypti artificially infected with Wolbachia pipientis was premature and did not properly assess the potential adverse impacts to the environment that may result from releases of Aedes aegypti artificially infected with a bacterium that is not native to the Aedes aegypti microbiome. EPA did not conduct a necessary rigorous assessment of the potential adverse impacts to human health and the environment that could result from uncontained release of Aedes aegypti containing this non native strain of bacteria. Of particular concern is the fact that horizontal gene transfer could result in Wolbachia effectively introducing over one thousand new genes into these modified Aedes aegypti. The potential genetic modification to Aedes aegypti that may result from use of the wAlbB Wolbachia bacterium is wholly undefined. Several studies have shown that horizontal gene transfer between Wolbachia and their insect hosts may result in gene transfers ranging from nearly the entire Wolbachia genome (>1 megabase) to short insertions (<500 base pairs) into various hosts [2-5], Effectively the Wolbachia IIT approach could introduce over one thousand new genes into Aedes aegypti with unknown consequences, and if they provide a positive selection to the mosquito in the environment this could result in novel strains pervading and spreading through the population. Critical questions about the Wolbachia Aedes aegypti that EPA did not answer prior to granting the EUP include the following: Can the Wolbachia mosquito escape the confined conditions in which it is reared? What is the likelihood that the Wolbachia mosquitoes will survive, reproduce, and disperse once released into the environment? Modelling has shown that conditions of lower competition can favor the spread of Wolbachia-infected females [6-8], In other words, as a mosquito population is reduced, or if a population is already low, the chances of Wolbachia invading the wild population are increased. 17cv1906 Sierra Club v. EPA - 6/22 Production ED 001523 00003114-00001  What are the potential impacts of the Wolbachia mosquitoes in the environment, including on humans? The whole genome of Wolbachia can transfer to a host genome, meaning a host mosquito could be transformed with over one thousand new genes with unpredictable results [2-5], Horizontal gene transfer (HGT) can transfer genes between Wolbachia and its host in Aedes aegypti [12] and several other mosquito species [13, 25], Therefore, Wolbachia can genetically transform its host with functional genes with currently unknown consequences. EPA did not require any investigation or risk assessment of these potential consequences. Moreover, widespread recombination occurs throughout the Wolbachia genome [14], increasing the likelihood of genes changing as the Wolbachia evolves. Moreover, temperature impacts Wolbachia-malaria interaction in mosquitoes suggesting impact of transfection might vary across diverse environments [17], Also, it has been shown that Wolbachia enhances West Nile virus infection in the mosquito Culex tarsalis. This introduces the possibility that the Wolbachia infection could spread to Culex populations in areas where West Nile virus is a concern [18], Similarly, Wolbachia can enhance malaria parasite infection in two genera of mosquitoes [19-21], What are the likely consequences for the surrounding environment, should the Wolbachia mosquitoes survive and establish in the environment? There is evidence that male age and overcrowding during development (i.e., under mass rearing conditions required to produce enough males for IIT to be effective) can reduce the cytoplasmic incompatibility effect in certain insects, rendering the males fertile [26] and able to spread the Wolbachia infection through surviving females. EPA should have required a study to assess whether this effect is likely to be seen in Aedes aegypti. EPA should have required additional analysis to assess interactions of Aedes aegypti, potential pathogens, and Wolbachia, to ensure that Wolbachia does not ultimately select for more dangerous pathogens in Aedes aegypti [22], EPA should have, but did not, evaluate these and other critical risk assessment questions before it granted the Wolbachia EUP extension, which allows the release of 14 million of these inadequately evaluated WoIbach/A-i nfectcd Aedes aegypti into the environment. As is demonstrated in the published literature referenced below, and as Oxitec and other commenters demonstrated on the record, releases of such massive numbers of these artificially infected Aedes aegypti could result in serious unreasonable adverse effects to the environment. 17cv1906 Sierra Club v. EPA - 6/22 Production ED 001523 00003114-00002 References 1) USEPA, Amendments, Extensions, and/or Issuances of Experimental Use Permits, 81 Federal Register 69059 October 5, 2016. 2) Dunning Hotopp, J.C., et al., Widespread lateral gene transfer from intracellular bacteria to multicellular eukaryotes. Science, 2007. 317: p. 1753-6. 3) Fenn, K., et al. Phylogenetic relationships of the Wolbachia of nematodes and arthropods. PLoS Pathog, 2006. 2: p. e94. 4) Hou, Q., et al, A case of horizontal gene transfer from Wolbachia to Aedes albopictus C6/36 cell line. Mob Genet Elements, 2014. 4: p. e28914. 5) Kondo, N., et al. Genome fragment of Wolbachia endosymbiont transferred to X chromosome ofhost insect. Proc Natl Acad Sci USA, 2002. 99: p. 14280-5. 6) Endersby, N.M. and A.A. Hoffmann, Effect of Wolbachia on insecticide susceptibility in lines of Aedes aegypti. Bulletin Entomol Res, 2013. 103(3): p. 269-77. 7) Hancock, P.A., S.P. Sinkins, and H.C.J. Godfray, Population dynamic models of the spread of Wolbachia. The American Naturalist, 2011. 177(3): p. 323-333. 8) Jansen, V.A.A., M. Turelli, and H.C.J. Godfray, Stochastic spread of Wolbachia. Proceedings ofthe Royal Society B: Biological Sciences, 2008. 275(1652): p. 2769-2776. 9) O'Connor, L., et al.. Open release of male mosquitoes infected with a Wolbachia biopesticide: field performance and infection containment. PLoS Negl Trop Dis, 2012. 6(11): p. el797. 10) Alphey, L., Natural and engineered mosquito immunity. Journal ofBiology, 2009. 8: p. 40. 11) Ye, Y.H., et al., Wolbachia reduces the transmission potential of dengue-infected Aedes aegypti. PLoS Negl Trop Dis, 2015. 9: e0003894. 12) Klasson, L., et al.. Horizontal gene transfer between Wolbachia and the mosquito Aedes aegypti. BMC Genomics, 2009. 10(1): p. 33. 13) Woolfit, M., et al.. An ancient horizontal gene transfer between mosquito and the endosymbiotic bacterium Wolbachiapipientis. Molecular biology and evolution, 2009. 26(2): p. 367-74. 14) Baldo, L., et al.. Widespread recombination throughout Wolbachia genomes. Mol Biol Evol, 2006. 23(2): p. 437-49. 15) Carrington, L.B., A.A. Hoffmann, and A.R. Weeks, Monitoring long-term evolutionary changes following Wolbachia introduction into a novel host: the Wolbachia popcorn infection in 17cv1906 Sierra Club v. EPA - 6/22 Production ED 001523 00003114-00003 Drosophila simulans. Proc Biol Sci, 2010. 277(1690): p. 2059-68. 16) Hughes, G.L., et al., Native microbiome impedes vertical transmission of Wolbachia in Anopheles mosquitoes. Proc Natl Acad Sci USA, 2014. 111: p. 12498-503. 17) Murdock, C.C., et al., Temperature alters Plasmodium blocking by Wolbachia. Sci Rep, 2014. 4: p. 3932. 18) Dodson, B.L., et al., Wolbachia enhances West Nile Virus (WNV) infection in the mosquito Culex tarsalis. PLoSNegl Prop Dis, 2014. 8(7): p. e2965. 19) Hughes, G.L., et al. Wolbachia can enhance Plasmodium infection in mosquitoes: implications for malaria control? PLoS Pathog, 2014. 10: p. el004182. 20) Hughes, G.L., et al. Wolbachia strain wAlbB enhances infection by the rodent malaria parasite Plasmodium berghei in Anopheles gambiae mosquitoes. Appl Environ Microbiol, 2012. 78: p. 1491-5. 21) Zele, F. et al., Wolbachia increases susceptibility to Plasmodium infection in a natural system. Proc Biol Sci, 2014. 281: p. 20132837. 22) Brelsfoard, C.L. and S.L. Dobson, Wolbachia-based strategies to control insect pests and disease vectors. AsPac J Mol Biol Biotechnol, 2009. 17(3): p. 55-63. 23) Werren, J.H., L. Baldo, and M.E. Clark, Wolbachia: master manipulators of invertebrate biology. Nat Rev Microbiol, 2008. 6(10): p. 741-51. 24) Marshall, J.L., Rapid evolution of spermathecal duct length in the Allonemobius socius complex of crickets: species, population and Wolbachia effects. PLoS One, 2007. 2(8): p. e720. 25) Baldo, L., et al.. Insight into the routes of Wolbachia invasion: high levels of horizontal transfer in the spider genus Agelenopsis revealed by Wolbachia strain and mitochondrial DNA diversity. MolEcol, 2008. 17(2): p. 557-69. 26) Yamada, R., et al., Male development time influences the strength of Wolbachia-induced cytoplasmic incompatibility expression in Drosophila melanogaster. Genetics, 2007. 177(2): p. 801-8. 17cv1906 Sierra Club v. EPA 6/22 Production ED 001523 00003114-00004