2007 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to create improved protocols for studies into the possible non-target effects of Bt crop production. Such protocols should improve the rigor of research into non-target effects and encourage collaboration among peer scientists in government, industry, and academic settings. 1b.Approach (from AD-416) We will use new and existing data to improve the protocols used to assess effects of Bt crops on non-target organisms. This will include recommendations on how, when, and what data should be collected, along with novel methods to analyze data. This includes some new field and laboratory research as well as new analyses of existing data. 3.Progress Report This report serves to document research conducted under a Specific Cooperative Agreement between ARS and the University of Maryland to conduct research related to an agreement with the USDA Biotechnology Risk Assessment Grants (BRAG) Program (60-3625-5-0599) entitled "Recommended Protocols for Field Evaluations of Non-Target Organisms in Bt Crops." Additional details can be found in the report for parent project #3625-22000-016-00D, Ecologically-Based Management of Insect Pests of Corn. This report is arranged according to the objectives in the original proposal. 1. Provide protocols for sampling non-target communities in Bacillus thuringiensis (Bt) corn, including appropriate (a) indicator taxa, (b) sample timing, and (c) replication. Progress on this objective can be found in the report for reimbursable agreement 3625-22000-016-18R. 2. Develop techniques to identify and rear abundant but currently indistinguishable larvae of non-target soil-dwelling beetles, and create laboratory methods for extended Bt exposure tests. Unlimited levels of Cry34/35 pollen have no adverse effects on the non-target beetles tested (Elaphropus xanthopus and Strigota ambigua), and both species prefer alternate prey to pollen. Tests are being conducted to quantify Cry34/35 consumption. Other lab feeding trials and field behavior studies also are underway to examine the role of minute ground beetles in corn. Manuscripts should be submitted for publication over the next 12 months. 3. Assess the suitability of different plot sizes for evaluating effects of cropping practices on non-target communities and specific indicator taxa. Progress on this objective can be found in the report for reimbursable agreement 3625-22000-016-18R. 4. Test approaches to combine and analyze existing data on the non-target effects of Bt corn to increase statistical power for detecting community and species-level treatment differences. Objectives 4 and 1 are related and utilize overlapping sets of data. The key difference is that Objective 4 focuses on optimal use of resources in multiple site (i.e., multiple field) studies, and explores pooling data from multiple studies to increase statistical power. Additional data sets (beyond those in Objective. 1)have been collected and analysis is planned for late 2007. Project Monitoring This project is monitored by frequent phone calls and emails. Also, to facilitate progress between the PIs, a meeting of PIs and industry stakeholders was held in Indianapolis during December 2006. Discussion at the meeting allowed the data sets for Objective 4 to be fully assembled and manuscript submission for Objective 1 to be completed.