2011 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to develop new fluorescent-based viral detection reagents for the study of Newcastle disease virus. 1b.Approach (from AD-416) Fluorescent molecular beacon-based probes targeted to the viral RNA will be used to analyse Newcastle Disease Virus replication. This approach allows for the analysis of the basic replication and assembly processes in four dimensions; involving space (x,y,z) and time. Initial studies will focus on the use of vaccine strain of NDV investigated in chicken embryo fibroblasts, while future studies will progress to mesogenic and velogenic strains for comparison. 3.Progress Report This research is related to inhouse objective 3: Biological and molecular characterization of emergent or previously undercharacterized Newcastle disease (NDV) isolates as the basis for extending the capabilities for epidemiology and the understanding of NDV ecology and evolution. Additional fluorescent molecular beacon-based probes targeted to the viral ribonucleic acid (RNA) to analyze NDV replication were tested. Initial evaluation using a vaccine strain of NDV in chicken embryo fibroblasts was completed; however it was found that some improvement was needed as the performance of the system has some limitations. As limited resources were available to continue development of new probes for Newcastle disease no additional work was performed on this system. Progress was monitored via regular email and telephone conversations with the cooperator.