To test the specificity of EHP in situ hybridization (ISH) and PCR
assays, samples of P. monodon showing clinical signs of cotton
shrimp disease were used, these shrimp were from Madagascar
in 2005 (fixed tissues) and 2006 (frozen tissues). P. vannamei
infected with an amoeba in the gill tissue, determined by histological
examination were also used, these were collected in 2014. For
PCR assays, hepatopancreas tissues or whole shrimp were preserved
in 95% ethanol or air dried and sent to our Lab; for ISH
assays, shrimp were fixed in Davidson’s AFA fixative.