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KMID : 0364820160520030313
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Korean Journal of Microbiology
2016 Volume.52 No. 3 p.313 ~ p.318
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Induction and resuscitation of viable but nonculturable Edwardsiella tarda
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Kang Nam-I
Kim Eun-Heui
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Abstract
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Bacteria in the viable but nonculturable (VBNC) state fail to produce colonies on routine bacteriological media, but are still
alive in the state of very low metabolic activity. The aim of the present study was to induce the VBNC state of the Edwardsiella tarda using sea water microcosm under starvation conditions at 10°C and to investigate resuscitation of the VBNC cells in temperatures changed from 10 to 25°C, with and without additives. E. tarda entered into the VBNC state within about 42?84 days of incubation in the microcosm. Throughout this period, the total cell counts as determined using acridine orange direct counting remained near the original inoculum level of ~108 cells/ml. The live cell counts measured with direct viable counting, on the other hands, declined to ~104 cells/ml. When the VBNC cells were incubated with addition of yeast extract, fish muscle extract or serum at 25°C, the ratios of resuscitated samples were 37%, 23%, and 37%, respectively. The characteristics of resuscitated E. tarda were consistent with those of the original E. tarda. When the resuscitated E. tarda were intraperitoneally injected into olive flounders, all fishes died within 5 days, indicating that the VBNC E. tarda might retain its pathogenic potential. Therefore, E. tarda under starvation conditions in the winter enter into the VBNC state and the VBNC E. tarda cells resuscitated at summer and autumn seawater temperature are considered to be pathogen continuously to olive flounder on the southern coast of Korea.
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KEYWORD
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Edwardsiella tarda, fish pathogen, resuscitation, viable but nonculturable (VBNC)
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