Growth Performance of a Newly Isolated and Culturable Thraustochytrid Strain from Sea Squirt Colonies

The world's oceans and seas host >100 known strains of thraustochytrids, a common group of marine eukaryotic unicellular protists, residing in diverse marine habitats, with many others to be isolated and cultivated. The thraustochytrids have become of considerable industrial interest due to health benefits gained from their high percentages of valuable bioactive compounds, revealing the needs for the isolation of new potential strains. Employing a recently developed isolation methodology (use of cell culture medium), we assess initial culture conditions and growth paces of newly isolated thraustochytrid cells (thraustochytrid sp. BSH), originated from the colonial tunicate Botryllus schlosseri, residing on Helgoland Island, Germany. Cells were cultivated under static versus agitated conditions, along with two inoculation sizes (0.5 x 10(6) and 1 x 10(6) cells/dish) and in three vessel types (35 mm Petri dishes and T25 and T75 flasks; containing 3, 5 and 15 mL medium, respectively). Cultures were observed under regular microscopy, confocal microscopy and H&E staining. While cells in all conditions grew fast, results revealed the superiority of agitated cultivation in T75 flasks inoculated with 0.5 x 10(6) cells/dish (6.41 +/- 1.91-fold increase/week). Further, 18S rDNA revealed high similarities (99.5-99.8) of strain BSH to two thraustochytrid strains from Monterey, California (USA), B. schlosseri colonies, elucidating a new understanding of these animals-protists associations.