2012 Annual Report 1a.Objectives (from AD-416): 1. Discover new biological control agents in Brazil from Diptera of medical and veterinary importance. 2. Evaluate biological control agents for Diptera and other vector species in Brazil and the United States. 1b.Approach (from AD-416): 1. Isolate, identify and develop new agents for the biological control of important Diptera such as Aedes aegypti, Culex and Anopheles spp. Information on the complex of natural control agents for each mosquito species will be catalogued and include but not restricted to mosquito pathogenic viruses, protozoa (microsporidia), bacteria, fungi and nematodes. New molecular probes will be developed and utilized for virus discovery. Technology developed by ARS will be employed to amplify and categorize newly found pathogens for further evaluation. 2. Conduct laboratory and field tests with new biological control agents to determine efficacy for the control of dipteran species that are vectors of diseases. Strategies to be investigated will be the use of biological control agents as part of an integrated mosquito management program to effectively control important vectors where specificity, recycling and resistance management are key objectives. 3.Progress Report: This research is related to in-house Objective 6: "Discover and characterize environmental predictors of the distribution of mosquitoes in order to assess the risk of invasive species and pathogen transmission. Apply to the development of methods and techniques to accurately assess mosquito population density, to deploy vector surveillance systems, and to detect exotic invasive species." This report covers the third year of a project between ARS researchers in Gainesville, FL and researchers in Brazil, Instituto Butantan. Brazilian mosquitoes are distinct but because of similar climatic conditions there is a high potential for invasion and establishment which could negatively impact U.S. agriculture. This project has evaluated control methods developed by ARS against Brazilian mosquitoes and isolation of new control agents to enhance remediation in the event of an invasion. This project could provide a model system for the protection of the U.S. from invasive mosquitoes in any part of the world. During this evaluation period, collaborative efforts have resulted in isolation of new mosquito pathogenic viruses in mosquitoes from Brazil. ARS has provided molecular probes to determine the identity of the isolated viruses and a manuscript was published.