The mission of U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services (WS) is to provide federal leadership and expertise in managing problems caused by wildlife. Approximately every 5 years, WS conducts a research needs assessment (RNA) to help to align research priorities at the National Wildlife Research Center (NWRC, the research arm of the WS program) with WS program and customer needs. In 2011, the WS Deputy Administrator solicited input from employees throughout the WS program and representatives from other federal agencies, all 50 state wildlife agencies, various livestock and agricultural commodity groups, and non-governmental organizations. Eighty-six federal employees from 36 states and the District of Columbia and 31 non-federal employees from 20 states responded to the RNA survey. Aviation safety, zoonotic diseases, livestock predation, and to a lesser degree protecting threatened and endangered (T&E) species and reducing crop depredations, were projected to be major areas of concern during the next 5 years. Invasive species, specifically feral swine, were one of the most frequently identified areas where research is needed. Development of nonlethal control methods and economic assessments were given a high priority. Many respondents wanted economic justification for their organizations or programs. Protection of aquaculture, property, and human safety, and development of vaccines and repellents were more localized concerns. Predation on livestock (especially cattle and sheep) and big game, waterfowl, and upland birds was a much bigger concern in the Western Region (WR) than the Eastern Region (ER). A higher percentage of WR respondents also anticipated being more involved in conflicts involving birds. Cormorants, beavers, deer, and especially vultures were of higher concern in the ER. State agency and private stakeholders most frequently identified either wildlife transmission of diseases or livestock depredation as their highest area of concern. State agency and private stakeholders most often identified development of more effective management techniques as their highest research priority. All respondents expressed a need for better economic information about the extent and nature of various human-wildlife conflicts. The results of this RNA, along with guidance from Congress and the WS Deputy Administrator and stakeholder input, will help establish WS research priorities.