Isolation of viable but nonculturable Vibrio cholerae O1 from environmental water samples in Kolkata, India, in a culturable state

Authors

  • Mitsutoshi Senoh,

    Corresponding author
    1. Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India
    Current affiliation:
    1. National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan
    • Correspondence

      Mitsutoshi Senoh, National Institute of Infectious Diseases, 4-7-1, Gakuen, Musashimurayama, Tokyo 208-0011, Japan. Tel:+81-42-561-0771; Fax:+81-42-561-7173; E-mail: senoh@nih.go.jp

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  • Jayeeta Ghosh-Banerjee,

    1. National Institute of Cholera and Enteric Diseases, Kolkata, India
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  • Tamaki Mizuno,

    1. Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India
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  • Sumio Shinoda,

    1. Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India
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  • Shin-ichi Miyoshi,

    1. Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan
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  • Takashi Hamabata,

    1. Research Institute, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan
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  • G. Balakrish Nair,

    1. Translational Health Science and Technology Institute, Haryana, India
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  • Yoshifumi Takeda

    1. Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India
    Current affiliation:
    1. Hideyo Noguchi Memorial Foundation, Inawashiro, Fukushima, Japan
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Abstract

Previously, we reported that viable but nonculturable (VBNC) Vibrio cholerae was converted into a culturable state by coculture with several eukaryotic cell lines including HT-29 cells. In this study, we found that a factor converting VBNC V. cholerae into a culturable state (FCVC) existed in cell extracts of eukaryotic cells. FCVC was nondialyzable, proteinase K-sensitive, and stable to heating at <60°C for 5 min. We prepared thiosulfate citrate bile salts sucrose (TCBS) plates with FCVC (F-TCBS plates). After confirming that VBNC V. cholerae O1 and O139 formed typical yellow colonies on F-TCBS plates, we tried to isolate cholera toxin gene-positive VBNC V. cholerae from environmental water samples collected in urban slum areas of Kolkata, India and succeeded in isolating V. cholerae O1 El Tor variant strains harboring a gene for the cholera toxin. The possible importance of VBNC V. cholerae O1 as a source of cholera outbreaks is discussed.

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