LUMINESCENCE-BASED DETECTION OF ACTIVITY OF STARVED AND VIABLE BUT NONCULTURABLE BACTERIA

Sarah Duncan, Lesley Anne Glover, Kenneth Stuart Killham, James Ivor Prosser

Research output: Contribution to journalArticle

105 Citations (Scopus)

Abstract

A naturally luminescent bacterium, Vibrio harveyi, and two bacteria, Escherichia coli and Pseudomonas fluorescens, which had been genetically marked with luminescence were starved in liquid medium at 4 and 30-degrees-C for 54 days. Total cell concentrations and concentrations of culturable and viable cells were determined by acridine orange staining, dilution plate counting, and direct viable counting, respectively, and population activity was measured by luminometry. V. harveyi became nonculturable but maintained viability during starvation at 4-degrees-C and maintained both culturability and viability at 30-degrees-C. In contrast, E. coli became viable but nonculturable during starvation at 30-degrees-C but not at 4-degrees-C. Luminescence of nonculturable cells of both strains, and culturable cells of V. harveyi, decreased to background levels during starvation. Luminescence of starved culturable cells of E. coli also fell below background levels but occasionally increased to detectable values. Viable, nonculturable forms of P. fluorescens were not detected at either temperature, and cells starved at 4-degrees-C showed no decrease in luminescence measured during incubation of samples at 25-degrees-C. Following incubation of late-log-phase cells with yeast extract and nalidixic acid, changes in light output directly paralleled changes in cell length, as observed during direct viable counting. Quantification of changes in luminescence following incubation of starved cells with yeast extract enabled measurement of the activity of both culturable and viable but nonculturable cells. Measurement of luminescence was significantly more sensitive, rapid, and convenient in quantifying activity following nutrient amendment than measurement of changes in cell length. Luminescence-based marker systems potentially provide a selective means of detecting the presence and activity of viable but nonculturable cells in the soil and freshwater environments, where indigenous luminescent populations are negligible, and enable assessment of the activity and environmental impact of such populations.

Original languageEnglish
Pages (from-to)1308-1316
Number of pages9
JournalApplied and Environmental Microbiology
Volume60
Issue number4
Publication statusPublished - Apr 1994

Keywords

  • GENETICALLY ENGINEERED MICROORGANISMS
  • VIBRIO-VULNIFICUS CELLS
  • ESCHERICHIA-COLI
  • PSEUDOMONAS-FLUORESCENS
  • MARINE-ENVIRONMENT
  • SOIL
  • BIOLUMINESCENCE
  • SURVIVAL
  • STATE
  • MICROSCOPY

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