Methodology: A previous survey (March 2015, before currently funded work) established that among 66 asteroids collected from Dutch Harbor, AK, none were positive for SSaDV. We again surveyed this population during aquarium specimen collection in September 2015 to determine the presence/load of SSaDV.
Sampling and analytical procedures: Animals were collected by hand or by SCUBA diver at 3 locations near Dutch Harbor, AK in September 2015. Body wall tissues were biopsied on site to determine the presence/load of SSaDV. Biopsy punches were immediately placed in 100% EtOH before transport back to the lab at Cornell University. At the same time, 68 live asteroids (n = 8 Pycnopodia helianthoides, n = 20 Solaster stimpsonii, n = 20 Evasterias troscheli, n = 20 Crossaster papposus) were collected by SCUBA diver at Dead Man’s Bend, placed into shipping containers, and then sent back to the lab at Cornell University. Unfortunately, all except 6 Evasterias troscheli and 3 Solaster stimpsonii died in transit due to Alaska Airlines and Fedex delays. Size was measured by measuring the diameter of the individual (longest arm to longest arm) visually using a ruler.
SSaDV abundance/load will be determined by qPCR following the approach of Hewson et al., 2014 (PNAS).