Easy Removal of Epiphytic Bacteria on Ulva (Ulvophyceae, Chlorophyta) by Vortex with Silica Sands

Macroalgae-associated bacteria play an important role in their algal hosts' biological processes. They are localized on surfaces of the host thalli, as well as between and even within algal cells. To examine the differences in community structures and functions between epi- and endo- bacteria, an effective approach for maximizing epiphyte removal from delicate seaweeds while retaining endophyte fidelity must be developed. In this study, a variety of surface sterilization methods for Ulva prolifera were compared, including mechanical, chemical, and enzymatical treatments. According to the results of scanning electron microscope (SEM) and denaturing gradient gel electrophoresis (DGGE) analysis, almost complete removal of epiphytic bacteria on Ulva was obtained simply by co-vortex of seaweeds with silica sands, causing minimal disturbance to endosymbionts when compared to previous published methods. In addition, the adaptability was also confirmed in additional U. prolifera strains and Ulva species with blade-like or narrow tubular thallus shapes. This easy mechanical method would enable the analysis of community composition and host specificity for Ulva-associated epi- and endo-bacteria separately.