Formulated aquafeed has been a primary driver for the growth of the aquaculture industry worldwide. However, assessment of the biosecurity of formulated aquafeed using unvalidated molecular diagnostics (i.e. polymerase chain reaction (PCR)- basedassay poses serious challenges, impacting the ability of shrimp feed manufacturers in the US and elsewhere to address safely and sustainably of the growing domestic and international aquaculture industry due to rejection of consignments.This proposal aims to develop standardized sample handling processes and PCR-based assays curated for testing aquafeed forwhite spot syndrome virus, the most important viral pathogen impacting shrimp aquaculture worldwide, and an emergent fungal pathogen,Enterocytozoon hepatopenaeiinfecting shrimp.The validated assay will be able to differntiate among aquafeed and feed ingredients that may contain fragment WSSV and EHP genome but poses no threat to shrimp health compared to those that may carry infectious pathogens.This will provide a much needed practical solution for the aquafeed industry facilitating the assurance of biosecurity of manufactured feeds until a crustacean cell line is available for screening infectious pathogens in aquafeed.