Detection of the kinetoplastid Azumiobodo hoyamushi, the causative agent of soft tunic syndrome, in wild ascidians Halocynthia roretzi.

The occurrence of soft tunic syndrome in wild populations of the ascidian Halocynthia roretzi was monitored by diving at 5 to 6 sites in Miyagi Prefecture in Japan in summer 2010 and 2011. These sites were located at varying distances from farming sites at which the disease had previously been detected. All dead ascidians were collected, and their tunics were examined for Azumiobodo hoyamushi, the causative agent of soft tunic syndrome, using 18S rRNA PCR. In both years, <1% of wild ascidians we observed (18 out of 2100 in 2010, and 30 out of 3100 in 2011) were dead. The flagellates were only detected in 8 out of 18 dead ascidians from 3 sites in 2010, and 4 out of 30 from 2 sites in 2011. Healthy ascidians were successfully experimentally infected with the disease by immersing tunic samples from diseased wild ascidians into the rearing water. When apparently healthy ascidians collected from the wild population were reared for 40 d using pathogen-free water, the tunics of some ascidians became softened. The flagellates were detected in these individuals, which were diagnosed with soft tunic syndrome. Our results suggest that soft tunic syndrome affects the wild population of ascidians in Japan.