These techniques include hatching eggs in spore-free water and rearing fry to the "ossification" stage in tanks or raceways.
Subsequently, animals were transferred to new, spore-free jars containing 80 mL of ADaM.
For infection, larvae were exposed to the spore-containing diet for three hours after which they were transferred to the spore-free diet until sampling.
Third, T. formosanum, originally described from Taiwan, was characterized by black ascocarp with apex warts and spores with free straight spine or irregular reticulum, which discriminated it them from other species (Figure 1g, h).
T. formosanum had a few morphological difference (black ascocarp with apex warts and spores with free straight spine) and a special host plant (Cyclobalanopsis glauca) and distribution (Taiwan island) that differed from T. indicum and, thus, was treated as a separate species.
The culture was then sonicated until >95% of the spores were free, as determined by phase-contrast microscopy.
Collectively, the latter four groups are sometimes referred to as pteridophytes, because each reproduces by spores liberated from dehiscent sporangia (free sporing).
Aquatic hyphomycetes are primarily dispersed in river currents as spores, either free-floating or trapped in foam.
After their release, the spores are "free-living" until the following year's cohort of juvenile hosts emerges.
When more than 95% free spores appeared, the spores were collected with cold, sterile, ultrapure water, centrifuged and washed extensively three times, before sedimentation through 20% and 50% Renografin (Bracco Sine, China) [10] and another three washes.
