title: Electrical Retrieval of Living Microorganisms from Cryopreserved Marine Sponges Using a Potential-Controlled Electrode.
abstract: The purpose of this study was to develop a novel electrical retrieval method (ER method) for living sponge-associated microorganisms from marine sponges frozen at -80 °C. A -0.3-V vs. Ag/AgCl constant potential applied for 2 h at 9 °C induced the attachment of the sponge-associated microorganisms to an indium tin oxide/glass (ITO) or a gallium-doped zinc oxide/glass (GZO) working electrode. The electrically attached microorganisms from homogenized Spirastrella insignis tissues had intact cell membranes and showed intracellular dehydrogenase activity. Dead microorganisms were not attracted to the electrode when the homogenized tissues were autoclaved for 15 min at 121 °C before use. The electrically attached microorganisms included cultivable microorganisms retrieved after detachment from the electrode by application of a 9-MHz sine-wave potential. Using the ER method, we obtained 32 phyla and 72 classes of bacteria and 3 archaea of Crenarchaeota thermoprotei, Marine Group I, and Thaumarchaeota incertae sedis from marine sponges S. insignis and Callyspongia confoederata. Employment of the ER method for extraction and purification of the living microorganisms holds potential of single-cell cultivation for genome, transcriptome, proteome, and metabolome analyses of bioactive compounds producing sponge-associated microorganisms.
authors.affiliation: Department of Marine Biodiversity Research, Japan Agency for Marine-Earth Science and Technology, 2-15 Natsushima-cho, Yokosuka, Kanagawa, 237-0061, Japan, skoyama@jamstec.go.jp.
authors.forename: Sumihiro
authors.initials: S
authors.lastname: Koyama
authors.forename: Shinro
authors.initials: S
authors.lastname: Nishi
authors.forename: Maki
authors.initials: M
authors.lastname: Tokuda
authors.forename: Moeka
authors.initials: M
authors.lastname: Uemura
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Ishikawa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Seya
authors.forename: Seinen
authors.initials: S
authors.lastname: Chow
authors.forename: Yuji
authors.initials: Y
authors.lastname: Ise
authors.forename: Yuji
authors.initials: Y
authors.lastname: Hatada
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Fujiwara
authors.forename: Taishi
authors.initials: T
authors.lastname: Tsubouchi
mesh_terms: D000818:Animals; D001105:Archaea; D001419:Bacteria; D004566:Electrodes; D011161:Porifera / Q000382:microbiology* / Q000502:physiology*