2. Materials and methods
2.1. Shrimp
E. hepatopenaei (EHP)-infected Penaeus stylirostris were
collected from Brunei in 2006. EHP-infected P. vannamei were
collected in shrimp ponds from Vietnam in 2014 and 2015. A population
of P. vannamei was sent from Thailand to our laboratory at
University of Arizona in 2014 for a quarantine purpose, these
shrimp were later diagnosed with EHP infection by histology, their
hepatopancreas, feces and tank water were sampled for PCR
analyses.
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.
2. Materials and methods2.1. ShrimpE. hepatopenaei (EHP)-infected Penaeus stylirostris werecollected from Brunei in 2006. EHP-infected P. vannamei werecollected in shrimp ponds from Vietnam in 2014 and 2015. A populationof P. vannamei was sent from Thailand to our laboratory atUniversity of Arizona in 2014 for a quarantine purpose, theseshrimp were later diagnosed with EHP infection by histology, theirhepatopancreas, feces and tank water were sampled for PCRanalyses.To test the specificity of EHP in situ hybridization (ISH) and PCRassays, samples of P. monodon showing clinical signs of cottonshrimp disease were used, these shrimp were from Madagascarin 2005 (fixed tissues) and 2006 (frozen tissues). P. vannameiinfected with an amoeba in the gill tissue, determined by histologicalexamination were also used, these were collected in 2014. ForPCR assays, hepatopancreas tissues or whole shrimp were preservedin 95% ethanol or air dried and sent to our Lab; for ISHassays, shrimp were fixed in Davidson’s AFA fixative.
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