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. 2018 May 29;9:1112. doi: 10.3389/fmicb.2018.01112

FIGURE 1.

FIGURE 1

Induction of viable but non-culturable (VBNC) state by Vibrio vulnificus in the artificial sea water on incubation at 4°C. (A) Microscopic pictures of wild-type V. vulnificus CMCP6 and pykA mutant strains after 0, 2, 4, and 11 days incubation at 4°C. Samples were stained with the Live/Dead BacLight Bacterial Viability Kit (Life Technologies). The two color fluorescence assay uses green nucleic acid stain that penetrates both live and dead cells, while the red propidium iodide stain is excluded from live cells. Representative data from three independent experiments are shown. (B) Total cell counts, viable cell counts, and plate counts were determined. Plate counts are given in cfu ml-1. ×, total cell counts of the wild-type V. vulnificus CMCP6 culture; ○, total cell counts of the pykA mutant culture; □, viable cell counts of the wild-type V. vulnificus CMCP6 culture; Δ, viable cell counts of the pykA mutant culture; ■, plate counts of the wild-type V. vulnificus CMCP6 culture; ▲, plate counts of the pykA mutant culture. The results are presented as the mean ± standard deviations of three independent measurements. Statistical significance for the VBNC cells on Day 11 was determined by Student’s t-test (∗P < 0.05, n = 3).