Measurement and interpretation of microbial adenosine tri-phosphate (ATP) in aquatic environments.

There is a widespread need for cultivation-free methods to quantify viability of natural microbial communities in aquatic environments. Adenosine tri-phosphate (ATP) is the energy currency of all living cells, and therefore a useful indicator of viability. A luminescence-based ATP kit/protocol was optimised in order to detect ATP concentrations as low as 0.0001 nM with a standard deviation of <5%. Using this method, more than 100 water samples from a variety of aquatic environments (drinking water, groundwater, bottled water, river water, lake water and wastewater effluent) were analysed for extracellular ATP and microbial ATP in comparison with flow-cytometric (FCM) parameters. Microbial ATP concentrations ranged between 3% and 97% of total ATP concentrations, and correlated well (R(2)=0.8) with the concentrations of intact microbial cells (after staining with propidium iodide). From this correlation, we calculated an average ATP-per-cell value of 1.75x10(-10)nmol/cell. An even better correlation (R(2)=0.88) was observed between intact biovolume (derived from FCM scatter data) and microbial ATP concentrations, and an average ATP-per-biovolume value of 2.95x10(-9)nmol/microm(3) was calculated. These results support the use of ATP analysis for both routine monitoring and research purposes, and contribute towards a better interpretation of ATP data.

[1]  Daniel Hoefel,et al.  Enumeration of water-borne bacteria using viability assays and flow cytometry: a comparison to culture-based techniques. , 2003, Journal of microbiological methods.

[2]  T. Egli,et al.  Correlations between total cell concentration, total adenosine tri-phosphate concentration and heterotrophic plate counts during microbial monitoring of drinking water , 2008 .

[3]  C. A. Davidson,et al.  Evaluation of two methods for monitoring surface cleanliness-ATP bioluminescence and traditional hygiene swabbing. , 1999, Luminescence : the journal of biological and chemical luminescence.

[4]  L. Stevenson,et al.  The contribution of bacteria to the total adenosine triphosphate extracted from the microbiota in the water of a salt-marsh creek☆ , 1981 .

[5]  John T. Wilson,et al.  Determination of Microbial Cell Numbers in Subsurface Samples , 1985 .

[6]  D. Coleman,et al.  Limitations of atp estimates of microbial biomass , 1984 .

[7]  T. Egli,et al.  Influence of size, shape, and flexibility on bacterial passage through micropore membrane filters. , 2008, Environmental science & technology.

[8]  R. Gourse,et al.  Relationship between Growth Rate and ATP Concentration in Escherichia coli , 2004, Journal of Biological Chemistry.

[9]  N. Yamaguchi,et al.  Improved Direct Viable Count Procedure for Quantitative Estimation of Bacterial Viability in Freshwater Environments , 2000, Applied and Environmental Microbiology.

[10]  Yingying Wang,et al.  The impact of industrial-scale cartridge filtration on the native microbial communities from groundwater. , 2008, Water research.

[11]  P. Martikainen,et al.  Biofilm formation in drinking water affected by low concentrations of phosphorus. , 2002, Canadian journal of microbiology.

[12]  Kasthuri Venkateswaran,et al.  ATP as a biomarker of viable microorganisms in clean-room facilities. , 2003, Journal of microbiological methods.

[13]  K. Imai,et al.  Enzymatic treatment to eliminate the extracellular ATP for improving the detectability of bacterial intracellular ATP. , 1997, Analytical biochemistry.

[14]  A. Magic-Knezev,et al.  Optimisation and significance of ATP analysis for measuring active biomass in granular activated carbon filters used in water treatment. , 2004, Water research.

[15]  T. H. Christensen,et al.  Distribution and Composition of Microbial Populations in a Landfill Leachate Contaminated Aquifer (Grindsted, Denmark) , 1999, Microbial Ecology.

[16]  R. Pridmore,et al.  ATP as a biomass indicator in eight North Island Lakes, New Zealand , 1984 .

[17]  K. Pedersen,et al.  Use of an ATP assay to determine viable microbial biomass in Fennoscandian Shield groundwater from depths of 3-1000 m. , 2007, Journal of microbiological methods.

[18]  Dick van der Kooij,et al.  Biofilm formation on surfaces of glass and Teflon exposed to treated water , 1995 .

[19]  D. Karl,et al.  Cellular nucleotide measurements and applications in microbial ecology. , 1980, Microbiological reviews.

[20]  H. Albrechtsen,et al.  Bulk water phase and biofilm growth in drinking water at low nutrient conditions. , 2002, Water research.

[21]  W D McElroy,et al.  The Energy Source for Bioluminescence in an Isolated System. , 1947, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Lundin,et al.  Detection of bacteriuria by luciferase assay of adenosine triphosphate , 1975, Journal of clinical microbiology.

[23]  Frederik Hammes,et al.  Rapid and direct estimation of active biomass on granular activated carbon through adenosine tri-phosphate (ATP) determination. , 2007, Water research.

[24]  Jordi Catalan,et al.  Suitability of Flow Cytometry for Estimating Bacterial Biovolume in Natural Plankton Samples: Comparison with Microscopy Data , 2007, Applied and Environmental Microbiology.

[25]  K. Y. Bφrshiem.  Cell volume to carbon conversion factors for a bacterivorous Monas sp. enriched from seawatr. , 1987 .

[26]  F. Azam,et al.  Dissolved ATP in the sea and its utilisation by marine bacteria , 1977, Nature.

[27]  B. K. Jensen.  ATP-related specific heterotrophic activity in petroleum contaminated and uncontaminated groundwaters , 1989 .

[28]  S. Günther,et al.  Limits of propidium iodide as a cell viability indicator for environmental bacteria , 2007, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[29]  V. Créach,et al.  Direct estimate of active bacteria: CTC use and limitations. , 2003, Journal of microbiological methods.

[30]  O. Holm‐Hansen.  DETERMINATION OF MICROBIAL BIOMASS IN OCEAN PROFILES1 , 1969 .

[31]  P E Stanley,et al.  A review of bioluminescent ATP techniques in rapid microbiology. , 1989, Journal of bioluminescence and chemiluminescence.

[32]  A. E. Greenberg,et al.  Standard methods for the examination of water and wastewater : supplement to the sixteenth edition , 1988 .

[33]  K. Pedersen,et al.  Numbers, biomass and cultivable diversity of microbial populations relate to depth and borehole-specific conditions in groundwater from depths of 4–450 m in Olkiluoto, Finland , 2008, The ISME Journal.

[34]  T. Egli,et al.  Isolation and characterization of low nucleic acid (LNA)-content bacteria , 2009, The ISME Journal.

[35]  G. Nebe-von Caron,et al.  Current and future applications of flow cytometry in aquatic microbiology. , 2000, FEMS microbiology reviews.

[36]  O. Köster,et al.  Flow-cytometric total bacterial cell counts as a descriptive microbiological parameter for drinking water treatment processes. , 2008, Water research.

[37]  B. Welt,et al.  An ATP-based method for monitoring the microbiological drinking water quality in a distribution network , 2003 .

[38]  T. Egli,et al.  Rapid, cultivation-independent assessment of microbial viability in drinking water. , 2008, Water research.

[39]  Frederik Hammes,et al.  Escherichia coli O157 can grow in natural freshwater at low carbon concentrations. , 2008, Environmental microbiology.

[40]  Yingying Wang,et al.  Quantification of the filterability of freshwater bacteria through 0.45, 0.22, and 0.1 microm pore size filters and shape-dependent enrichment of filterable bacterial communities. , 2007, Environmental science & technology.

[41]  B. Riemann.  The occurrence and ecological importance of dissolved ATP in fresh water , 1979 .

[42]  J. Gasol,et al.  Seasonal Variations in the Contributions of Different Bacterial Groups to the Uptake of Low-Molecular-Weight Compounds in Northwestern Mediterranean Coastal Waters , 2007, Applied and Environmental Microbiology.

[43]  Thomas Egli,et al.  Flow-cytometric study of vital cellular functions in Escherichia coli during solar disinfection (SODIS). , 2006, Microbiology.