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Induction and Resuscitation of the Viable but Nonculturable State in a Cyanobacteria-Lysing Bacterium Isolated from Cyanobacterial Bloom

  • Environmental Microbiology
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Abstract

The viable but nonculturable (VBNC) state has been found to be a growth strategy used by many aquatic pathogens; however, few studies have focused on VBNC state on other aquatic bacterial groups. The purpose of this study was to explore the VBNC state of cyanobacteria-lysing bacteria and the conditions that regulate their VBNC state transformation. Three cyanobacteria-lysing heterotrophic bacterial strains (F1, F2 and F3) were isolated with liquid infection method from a lake that has experienced a cyanobacterial bloom. According to their morphological, physiological and biochemical characteristics and results of 16SrDNA sequence analysis, F1, F2 and F3 were identified as strains of Staphylococcus sp., Stappia sp. and Microbacterium sp., respectively. After being co-cultured with the axenic cyanobacterium, Microcystis aeruginosa 905, for 7 days, strains F1, F2 and F3 exhibited an inhibition effect on cyanobacterial growth, which was expressed as a reduction in chlorophyll concentration of 96.0%, 94.9% and 84.8%, respectively. Both autoclaved and filtered bacterial cultures still showed lytic effects on cyanobacterial cells while centrifuged pellets were less efficient than other fractions. This indicated that lytic factors were extracelluar and heat-resistant. The environmental conditions that could induce the VBNC state of strain F1 were also studied. Under low temperature (4°C), distilled deionized water (DDW) induced almost 100% of F1 cells to the VBNC state after 6 days while different salinities (1%, 3% and 5% of NaCl solution) and lake water required 18 days. A solution of the cyanobacterial toxin microcystin-LR (MC-LR) crude extract also induced F1 to the VBNC state, and the effect was stronger than DDW. Even the lowest MC-LR concentration (10 μg L−1) could induce 69.7% of F1 cells into VBNC state after 24 h. On the other hand, addition of Microcystis aeruginosa cells caused resuscitation of VBNC state F1 cells within 1 day, expressed as an increase of viable cell number and a decrease of VBNC ratio. Both VBNC state and culturable state F1 cells showed lytic effects on cyanobacteria, with their VBNC ratio varying during co-culturing with cyanobacteria. The findings indicated that VBNC state transformation of cyanobacteria-lysing bacteria could be regulated by cyanobacterial cells or their toxin, and the transformation may play an important role in cyanobacterial termination.

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Acknowledgments

We thank the anonymous reviewers for thoroughly reading the paper, providing thoughtful comments. This work was been supported by the National Natural Science Foundation of China (nos. 31070323 and 30770340), Major Science and Technology Program for Water Pollution Control and Treatment (nos. 2009ZX07423-003 and 2009ZX07101-011-03), Shenzhen Grant Plan for Science and Technology to Z. Hu, International Mobility Fund Grant of Royal Society of New Zealand (IMF10-A61) to W.L.Wihte and Z.Hu, and a China Postdoctoral Science Foundation (no. 20110490924) to H. Chen.

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Authors and Affiliations

  1. Shenzhen Key Laboratory of Marine Bioresource and Eco-environmental Science, College of Life Science, Shenzhen University, Shenzhen, 518060, China

    Huirong Chen, Lixian Fu, Lingxi Luo, Jun Lu & Zhangli Hu

  2. Department of Interdisciplinary Studies, Faculty of Health and Envinronmental Sciences, Auckland University of Technology, Auckland, New Zealand

    Jun Lu & W. Lindsey White

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  1. Huirong Chen
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  2. Lixian Fu
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Correspondence to Zhangli Hu.

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Chen, H., Fu, L., Luo, L. et al. Induction and Resuscitation of the Viable but Nonculturable State in a Cyanobacteria-Lysing Bacterium Isolated from Cyanobacterial Bloom. Microb Ecol 63, 64–73 (2012). https://doi.org/10.1007/s00248-011-9928-2

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  • DOI: https://doi.org/10.1007/s00248-011-9928-2

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