caa a22 4500 445892528 CHVBK 20180317145621.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s10750-010-0488-0 doi (NATIONALLICENCE)springer-10.1007/s10750-010-0488-0 Axenic culture of Brachionus plicatilis using antibiotics [Elektronische Daten] [Koushirou Suga, Yukari Tanaka, Yoshitaka Sakakura, Atsushi Hagiwara] The rotifer Brachionus plicatilis culture is composed of complex microcosms including bacteria, protozoans, algae, and fungi. Previous studies reported methods to establish axenic rotifer cultures, but further refinement of these techniques is needed, for molecular biological research which requires pure culture to isolate nucleic acids from rotifers only. In order to render rotifer culture axenic, we tested five antibiotics: ampicillin (Amp), chloramphenicol (Cp), kanamycin (Km), nalidixic acid (Na), and streptomycin (Sm) at 30-100μg/ml. Except for Cp, which reduces rotifer reproduction, all other antibiotics at the tested concentrations did not affect rotifer reproduction or show any toxic effects. A rotifer disinfection method was finally established by treating the resting eggs with 0.25% (w/v) sodium hypochlorite (NaOCl) for 3min, washing with sterilized sea water, and then exposing the neonates to an Amp, Km, Na, and Sm mixture. Using four nutrient media, we confirmed that this protocol renders the rotifer culture bacterial and fungus free. The axenic rotifer culture generated here is useful not only for genetic analysis of Brachionus plicatilis, but for studying the rotifer life cycle without bacterial influence. Springer Science+Business Media B.V., 2010 Brachionus plicatilis nationallicence Antibiotic nationallicence Bacteria free nationallicence Resting egg disinfection nationallicence Suga Koushirou Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut Tanaka Yukari Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut Sakakura Yoshitaka Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut Hagiwara Atsushi Graduate School of Science and Technology, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut Hydrobiologia Springer Netherlands 662/1(2011-03-01), 113-119 0018-8158 662:1<113 2011 662 10750 https://doi.org/10.1007/s10750-010-0488-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10750-010-0488-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Suga Koushirou Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Tanaka Yukari Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Sakakura Yoshitaka Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 700 1- Hagiwara Atsushi Graduate School of Science and Technology, Nagasaki University, Bunkyo 1-14, 852-8521, Nagasaki, Japan aut NATIONALLICENCE 773 0- Hydrobiologia Springer Netherlands 662/1(2011-03-01), 113-119 0018-8158 662:1<113 2011 662 10750 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer