Pycnopodia helianthoides sea star wasting disease exposure experiment- Control DB
Pycnopodia helianthoides (radius 160.5 +/- 30.9 mm) were collected from four sites in Washington State: Langley (LA) (48.038, -122.404); Dabob Bay (DB) (47.813, -122.820); Port Hadlock Marina (PH) (48.030, -122.745); Friday Harbor (FH) (48.545, -123.012). Sites had no reports of SSWD infection at time of collection. Individuals were inoculated with tissue homogenate prepared from the tube feet, dermal tissue, and coelomic fluid of three P. helianthoides with clinical signs of SSWD. The tissue was ground with a mortar and pestle, then placed in a Stomacher with 10 mL seawater, and centrifuged at 1000 x g for 5 min at 4°C. Organisms injected with the heat-killed homogenate comprised the control group. The treated group were injected with viral-sized fraction homogenate that was syringe-filtered through a 0.22 µm polyethylsulfone filter. Libraries were prepared using the Illumina TruSeq RNA Sample Preparation kit according to the manufacturer’s protocol