**UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460** 

OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION

**SEPT 17 2010** MEMORANDUM

SUBJECT: BPPD Response to ABSTC* Comments on Proposed Terms and Conditions for 2010 Expiring Bt Corn Registrations. Data Required to Register Plant-Incorporated Protectants: Aquatic Invertebrates and Orius Testing Issues

FROM: Zigfridas Vaituzis, Senior Scientist (signed 9-17-10) Microbial Pesticides Branch Biopesticides and Polluti on Prevention Division, 7511P

TO: Mike Mendelsohn, Senior Re gulatory Action Leader Microbial Pesticides Branch Biopesticides and Pollu tion Prevention Division, 7511P

*The ABSTC members are: Ba yer CropScience; Dow AgroSciences LLC; Pioneer Hi-Bred International, a DuPont Company; Monsan to Company; and Syngenta Seeds, Inc

I. AQUATIC INVERTEBRATE ISSUE

Background:

As a result of a recently published study (Rosi-Ma rshall, et al. 2007) showing weight reduction in shredding caddis flies exposed to anti-lepidopteran Cry1A protein corn litter in a laboratory setting, EPA required the registrants of PIP crops to submit additional dietary Daphnia magna studies, or studies on the effects on an aquatic invertebrate representing a leaf shredder functional group in headwater streams. These data were to be submitted by January 31, 2012.

Current Status:

Numerous recent peer-reviewed studies performe d as a result of the Rosi-Marshall et al. publication identified several issues with the sc ientific merit and relevance to environmental risk assessment of the original study (Swan et al. 2009, Jensen et al. 2010, summarized by Beachy et al.2008, Parrott 2008, and Wolt and Peters on 2010). It appears that the adverse effects in the Rosi-Marshall la boratory study were not caused by toxicity of Cry1A, but were due to other differences betw een the plant test substances (Jensen et al. 2010) . Field studies did not indicate any reduced abundance of Trichopter a in streams containing residues of Cry1A maize ( Chambers et al. (2007). In addition, the data in Rosi-Marshall et al. (2007) corroborate a lack of risk to aquatic organisms in realistic environmental conditions, as presented in Chambers etPage 2 of 4 al. (the authors of Chambers et al. are the same research group that produced the Rosi-Marshall et al. 2007 paper). Finally, post-harvest crop residue was identified as the most likely route of exposure (Carstens et al. 2010), however aquatic exposure to biotech crops has been shown to be limited temporally and spatially with low to neglig ible exposure concentrations of Cry proteins in post-harvest crop tissues (Swan et al. 2009, Griffiths et al. 2009, Jensen et al. 2010, Wolt and Peterson 2010, Carstens et al. 2010) Similar e xposure characterization for pending biotech crops during the problem formulation stage, on a case-by-case basis, may likely indicate a lack of need for hazard testing to assess aquatic risks in many instances (ILSI-CERA 2009, Carstens et al. 2010).

EPA Conclusion :

The caddis fly matter is no longer an issue. Nu merous peer-reviewed studies performed since the original publication claiming a possible environmental hazard, (as a result of Cry1A protein causing weight reduction to caddis f lies in a laboratory study), show that the anticipated problem does not exist. The studies show a lack of toxi city and a lack of si gnificant exposure in the environmental setting. Therefore the requirement fo r additional aquatic invertebrate studies to assess hazard to aquatic shredder species is waived fo r the existing Cry protein PIP registrations.

II. ORIUS DATA REQUIREMENT

Background:

The August 2002 FIFRA SAP (EPA-SAP. November 6, 2002) recommended that EPA expand the non-target insect te sting to include genera list predators such as the insidious flower bug Orius insidiosus (sometimes referred to as the minute pirate bug). As a result EPA required the registrants of expiring registrations to submit dietary studies with Orius insidiosus

. These data were to be submitted by January 31, 2012.

Current Status:

For commercialized crops containing insecticidal Cry proteins from Bacillus thuringiensis active against Lepidoptera and Coleoptera species, spectrum data indicate that toxicity is restricted only to insect larvae in these two Orde rs and biologically meaningful activity against Orius (a hemipteran insect) by anti lepidopteran and anti-coleopter an Cry ptoeins is low to nonexistent ( Duan et al. 2006, Raybould et al. 2007) . In addition, field data performed on commercialized events have not indicated adverse impacts on hemipteran pests within agronomic fields. (Sanvido et al.,2007, Marvier, et al., 2007). Consideration of such spectrum and field data would indicate a lack of need for further testing of with Orius or other hemipterans for existing Cry protein containing crops. Furthermore, commer cial laboratories performing the testing encountered difficulties with rearing a nd maintaining acceptable control mortality levels in Orius tests (EPA MRID 474367-01. 2008). Also, at the present time there are no independent laboratories performing the assay. The Springbor n Laboratories, an independent contract research organization and co-autho r on the Duan et al. (2007) paper, is no longer performing this assay. Therefore, ABSTC reque sts that requirements for Orius testing be suspended for currently registered Bt Cry protein crops.Page 3 of 4 EPA Conclusion:

The EPA has made a good faith effort to comply with the 2002 FIFRA SAP recommendation to expand non-target insect testing to include generalist predator speci es such as the minute pirate bug Orius insidiosus. The regulated industry responded at considerable expenditure of resources to comply with the requirement. However, due to lack of contract labor atories performing the testing, difficulty in obtaining and rearing of Orius, and repeated failure to maintain acceptable control mortality levels the testing has not been sati sfactory. Therefore the requirement for additional Orius studies to assess hazard to generalist pred ator species is waived for the existing Cry protein PIP registrations, and th at testing of active ingredient s other than Cry proteins be required only after development and va lidation of testing protocols.

References

Beachy, RN, NV Federoff, RB Goldberg, and A McHughen. 2008. The burden of proof: a response to Rosi-Marshall et al. Proc Natl Acad Sci USA 105: E9.

Carstens, K.L., Anderson, J.A., Bachman, P., DeShrijver, A., Dively, G. Federici, B. Hamer, M., Gielkens, M., Jensen, P., Lamp, W., Raushen, S., Ridley, G., Romeis, J., Waggoner, A. 2010. Genetically Modified Crops and Aquatic Ecosystems: Considerations for Environmental Risk Assessment and Non-target Organism Testing. In preparation.

Chambers, C., Whiles, M.R. Griffiths, N.A., Evans-White, M.A., Rosi-Marshall, E.J., Tank, J.L., and Royer, T.V. 2007. Assessing the impacts of transgenic Bt corn detritus on macroinvertebrate communities in agricultural streams. North American Benthological Society 55 th Annual Meeting. June 7, 2007. Columbia, SC. http://nabs.confex.com/nabs/2007/techprogram/P1698.HTM

Duan, J.J., Paradise, M.S., Lundgren, J.G., Bookout, J.T., Jiang, C., and Wiedenmann, R.N. 2006. Assessing nontarget impacts of Bt corn resistant to corn rootworms: tier-1 testing with larvae of Poecilus chalcites (Coleoptera: Carabidae). Environ. Entomol . 35:135-142.

Duan, J.J., Huesing, J. Teixeira, D. 2007. Development of tier-I toxicity assays for Orius insidiosus (Heteroptera: Anthocoridae) for assessing the ri sk of plant-incorporated protectants to nontarget heteropterans. Environ. Entomol. 36(4):982-8.

EPA-SAP. November 6, 2002. Corn rootworm plant- incorporated protecta nt insect resistance management and non-target insect issues. Transmittal of meeting minutes of the FIFRA Scientific Advisory Panel Meeting held A ugust 27-29, 2002 at the Marriott Crystal City Hotel, Arlington, VA.

EPA. MRID 474367-01. 2008. Evaluation of Potentia l Dietary Effects of Cry34/35Ab1 Protein on Insidious Flower Bugs, Orius insidiosus (Hemiptera: Anthocoridae), Study No. 13855.6105, Springborn Smithers Laboratories, 7 90 Main Street, Wareham, MA 02571- 1037.Page 4 of 4 ILSI-CERA (International Life Sciences Institute Research Foundation, Center for Environmental Risk Assessment). Problem Formulation of Biotech Crops and Aquatic Ecosystems. 1156
15 th Street, NW, Washington, DC 20005. October 13-15, 2009.

Jensen, P.D., G.P. Dively, C.M. Swan, and W.O. Lamp. 2010. Exposure and non-target effects of transgenic Bt corn debris in streams. Environ. Entomol. 39(2):707-714.

Marvier, M., C. McCreedy, J. Regetz, and P Ka reiva. 2007. A Meta-Analysis of Effects of Bt Cotton and Maize on Nontarget Invertebra tes. Scince, Vol 316 8 June 2007, 1475-1477.

Parrott, W. 2008. Study of Bt impact on caddisflies overstates its conclusions: Response to RosiMarshall et al. Proc. Natl. Acad. Sci. USA 105(7): E10.

Raybould, A., Stacey, D., Vlachos, D., Graser, G., Li , X., and Joseph, R. 2007. Non-target organism risk assessment of MIR604 maize expressing mCry3A for control of corn rootworm. J. Appl. Entomol. 131(6): 391-399.

Rosi-Marshall, E.J., Tank, J.L., Royer, T.V., Whiles, M.R., Evans-White, M., Chambers, C., Griffiths, N.A., Pokelsek, J., and Stephen, M.L. 2007. Toxins in transgenic crop byproducts may affect headwater stream ecosystems. Proc. Natl. Acad. Sci. USA 104:16204–16208.

Sanvido,O., Romeis, J., Bigler, F. 2007. Ecological Impacts of Genetically Modified Crops: Ten Years of Field Research a nd Commercial Cultivation. A dv Biochem Engin/Biotechnol 107: 235–278

Swan, C.M., P.D. Jensen, G.P. Dively, and W.O. Lamp. 2009. Processing of transgenic crop residues in stream ecosystems. J. App. Ecol. 46:1304-1313.

Wolt, J.D. and Peterson, R.K.D. 2010. Prosp ective formulation of environmental risk assessments: Probabilistic screening for Cr y1A(b) maize risk to aquatic insects. Ecotoxicol. Environ. Saf. Epub.