Induction and resuscitation of viable nonculturable Arcobacter butzleri cells

Abstract

Two strains of Arcobacter butzleri, ATCC 49616 and an environmental isolate, became nonculturable in seawater microcosms at 4 degrees C by 20 days and at room temperature by 14 days. Nonculturable cells were viable for up to 270 days of incubation in microcosms. Resuscitation of A. butzleri cells from microcosms at both temperatures was achieved 9 days after nutrient addition.

FIG. 1.

Survival curves of Arcobacter butzleri ATCC 49616 in filter-sterilized seawater microcosms at room temperature (A) and 4°C (B). Culturable cells (numbers of CFU ml⁻¹) (□) were counted on AM agar plates; viability (numbers of cells ml⁻¹) (♦) was determined by a Live/Dead BacLight viability assay. Each point represents the mean of two repeated and independent experiments, and error bars for culturable cells indicate standard deviations of the distribution of the data.

FIG. 2.

Survival curves of Arcobacter butzleri strain Ap-6 in filter-sterilized seawater microcosms at room temperature (A) and 4°C (B). Culturable cells (numbers of CFU ml⁻¹) (□) were counted on AM agar plates; viability (numbers of cells ml⁻¹) (♦) was determined by a Live/Dead BacLight viability assay. Each point represents the mean of two repeated and independent experiments, and error bars for culturable cells indicate standard deviations of the distribution of the data.

FIG. 3.

FISH micrographs of Arcobacter butzleri ATCC 49616 cells at inoculation time (A) and after 270 days of incubation (B) in a filter-sterilized seawater microcosm at room temperature.