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Characterization and resuscitation of viable but nonculturable Vibrio alginolyticus VIB283

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Abstract

The aim of this study was to investigate the viable but nonculturable (VBNC) state of the bacterium. Vibrio alginolyticus VIB283 was cultured in sterilized seawater microcosm at 4°C. Culturability of the cells in the microcosm was monitored by spread plate count (PC) on 2216E agar, PCs declined to undetectable levels (<0.1 CFU/ml) within 90 days. Total cell counts remained constant throughout the period as determined by acridine orange direct count (AODC). The direct viable counts, on the other hand, declined from 1010 to 109 CFU/ml active cells and remained fairly constant at this level by direct viable count (DVC), which indicated that a large population of cells entered into the VBNC state. The VBNC cells could be resuscitated by temperature upshift with and without the presence of nutrition. The resuscitated time were 16 h and 8 days respectively. The resuscitation was not achieved in chick embryos. The morphology of the VBNC, normal and resuscitated cells was studied with scanning electron microscope and flow cytometry. The cells changed from rod or arc to coccoid and decreased in size when entered into the VBNC state. The resuscitated and the normal cells had almost no morphological differences.

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Abbreviations

AODC:

Acridine orange direct count

DVC:

Direct viable count

VBNC:

Viable but nonculturable

PC:

Plate count

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Acknowledgements

This work was supported by a grant from the National Natural Science Foundation of China 30371108 and National High-Tech. R & D Program 2003AA622070.

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Authors and Affiliations

  1. Department of Marine Biology, College of Marine Life Sciences, Ocean University of China, 5# Yushan Road, Qingdao, 266003, People’s Republic of China

    Meng Du, Jixiang Chen, Xiaohua Zhang, Aijuan Li & Yun Li

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  1. Meng Du
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  2. Jixiang Chen
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  3. Xiaohua Zhang
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  4. Aijuan Li
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  5. Yun Li
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Correspondence to Jixiang Chen.

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Communicated by Jorge Membrillo-Hernández.

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Du, M., Chen, J., Zhang, X. et al. Characterization and resuscitation of viable but nonculturable Vibrio alginolyticus VIB283. Arch Microbiol 188, 283–288 (2007). https://doi.org/10.1007/s00203-007-0246-5

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