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Axenic culture of Brachionus plicatilis using antibiotics

  • ROTIFERA XII
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Abstract

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 30–100 μg/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 3 min, 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.

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Acknowledgments

A part of this research was supported by a Grant in Aid for Scientific Research (B), 2009–2011, No. 21380125, from the Ministry of Education, Culture, Sports, Science and Technology of Japan, to A. H. We would like to thank Dr. Helen S. Marcial for her review and comments.

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Authors and Affiliations

  1. Faculty of Fisheries, Nagasaki University, Bunkyo 1-14, Nagasaki, 852-8521, Japan

    Koushirou Suga, Yukari Tanaka & Yoshitaka Sakakura

  2. Graduate School of Science and Technology, Nagasaki University, Bunkyo 1-14, Nagasaki, 852-8521, Japan

    Atsushi Hagiwara

Authors
  1. Koushirou Suga
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  2. Yukari Tanaka
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  3. Yoshitaka Sakakura
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  4. Atsushi Hagiwara
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Correspondence to Koushirou Suga.

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Guest editors: N. Walz, R. Adrian, J.J. Gilbert, M.T. Monaghan, G. Weithoff & H. Zimmermann-Timm / Rotifera XII: New aspects in rotifer evolution, genetics, reproduction, ecology and biogeography

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Suga, K., Tanaka, Y., Sakakura, Y. et al. Axenic culture of Brachionus plicatilis using antibiotics. Hydrobiologia 662, 113–119 (2011). https://doi.org/10.1007/s10750-010-0488-0

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