Applied Microbiology and Biotechnology

, Volume 102, Issue 1, pp 367–375 | Cite as

Distinguishing between metabolically active and dormant bacteria on paper

  • Stephanie A. Hice
  • Miguel C. Santoscoy
  • Michelle L. Soupir
  • Rebecca CademartiriEmail author
Applied microbial and cell physiology


Switching between metabolically active and dormant states provides bacteria with protection from environmental stresses and allows rapid growth under favorable conditions. This rapid growth can be detrimental to the environment, e.g., pathogens in recreational lakes, or to industrial processes, e.g., fermentation, making it useful to quickly determine when the ratio of dormant to metabolically active bacteria changes. While a rapid increase in metabolically active bacteria can cause complications, a high number of dormant bacteria can also be problematic, since they can be more virulent and antibiotic-resistant. To determine the metabolic state of Escherichia coli and Salmonella Typhimurium, we developed two paper-based colorimetric assays. The color changes were based on oxidoreductases reducing tetrazolium salts to formazans, and alkaline phosphatases cleaving phosphates from nitrophenyl phosphate salt. Specifically, we added iodophenyl-nitrophenyl-phenyl tetrazolium salt (INT) and methylphenazinium methyl sulfate to metabolically active bacteria on paper and INT and para-nitrophenyl phosphate salt to dormant bacteria on paper. The color changed in less than 60 min and was generally visible at 103 CFU and quantifiable at 106 CFU. The color changes occurred in both bacteria, since oxidoreductases and alkaline phosphatases are common bacterial enzymes. On one hand, this feature makes the assays suitable to a wide range of applications, on the other, it requires specific capture, if only one type of bacterium is of interest. We captured Salmonella or E. coli with immobilized P22 or T4 bacteriophages on the paper, before detecting them at levels of 102 or 104 CFU, respectively. Determining the ratio of the metabolic state of bacteria or a specific bacterium at low cost and in a short time, makes this methodology useful in environmental, industrial and health care settings.


Bacteriophage Escherichia coli Salmonella Metabolic state Detection Paper 



The authors thank Abigail Meyer for her contribution to the colorimetric assays.


This research was funded in part by the Water Environment Research Foundation (WERF) (No. U4R13), the Leopold Center for Sustainable Agriculture (LCSA, No. ESP2014-02), the Midwest Dairy Association (MDA), and by the Center for Health Effects of Environmental Contamination (CHEEC).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2017_8604_MOESM1_ESM.pdf (280 kb)
ESM 1 (PDF 280 kb).


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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.Department of Food Science and Human NutritionIowa State UniversityAmesUSA
  2. 2.Department of Chemical and Biological EngineeringIowa State UniversityAmesUSA
  3. 3.Department of Agricultural and Biosystems EngineeringIowa State UniversityAmesUSA
  4. 4.Department of Material Science and EngineeringIowa State UniversityAmesUSA

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