SURVEILLANCE OF AQUACULTURE HATCHERY WATER FOR FISH PATHOGEN, IHNV

INFECTIOUS HEMATOPOIETIC NECROSIS (IHN) VIRUS IS A PATHOGENIC AGENT WHICH INFECTS TROUT AND SALMON WITH A HIGH RATE OF FATALITY. THE IHN VIRUS IS WATERBORNE AND HAS BEEN INDICTED IN EPIZOOTIC OUTBREAKS OF DISEASE IN THE STATES OF WASHINGTON, OREGON AND CALIFORNIA. THE ECONOMIC LOSS TO THEAQUACULTURE INDUSTRY DUE TO THIS ONE VIRUS IS BELIEVED TO BE IN THE MILLIONS OF DOLLARS ANNUALLY. AT THE PRESENT TIME, NO ADEQUATE METHOD EXISTS FOR MONITORING THE IHN VIRUSIN HATCHERY WATER OR FOR TRACING THE SPEED OF THIS DISEASE. THE OBJECTIVE OF THIS RESEARCH PROJECT IS TO DEVELOP SUCH A DETECTION METHOD. THE IHN VIRUS IS KNOW TO BE TRANSMISSIBLE THROUGH WATER WHERE IT SHOULD BE DETECTABLE EVEN AT LOW CONCENTRATIONS, USING THE TECHNOLOGY OF NUCLEIC ACID AMPLIFICATION COUPLED TO DETECTION WITH DNA PROBES. DURING PHASE I, THE FEASIBILITY OF SUCH A METHOD WILL BE EXAMINED BY: 1) IDENTIFYING PROBE SEQUENCES WHICH DEFINE AMPLIFIABLE REGIONSSPECIFIC TO THE VIRUS; AND 2) ESTABLISHING A PROCEDURE TO COLLECT AND CONCENTRATE VIRUSES FROM WATER SAMPLES WHICH PERMITS ISOTHERMAL AMPLIFICATION OF VIRAL SEQUENCES. COUPLING THE AMPLIFICATION AND THE SAMPLE PREPARATION WITH ANON-RADIOACTIVE DETECTION METHOD FOR USE AS AN ON-SITE AQUACULTURE DIAGNOSTIC TEST WILL BE A MAJOR TASK OF PHASE II.