Effect of treated-sewage contamination upon bacterial energy charge, adenine nucleotides, and DNA content in a sandy aquifer on Cape Cod

Changes in adenylate energy charge (ECA) and in total adenine nucleotides (A(T) and DNA content (both normalized to the abundance of free-living, groundwater bacteria) in response to carbon loading were determined for a laboratory-grown culture and for a contaminated aquifer. The latter study involved a 3-km-long transect through a contaminant plume resulting from continued on-land discharge of secondary sewage to a shallow, sandy aquifer on Cape Cod, Mass. With the exception of the most contaminated groundwater immediately downgradient from the contaminant source, DNA and adenylate levels correlated strongly with bacterial abundance and decreased exponentially with increasing distance downgradient. ECAS (0.53 to 0.60) and the ratios of ATP to DNA (0.001 to 0.003) were consistently low, suggesting that the unattached bacteria in this groundwater study are metabolically stressed, despite any eutrophication that might have occurred. Elevated ECAS (up to 0.74) were observed in glucose-amended groundwater, confirming that the metabolic state of this microbial community could be altered. In general, per-bacterium DNA and ATP contents were approximately twofold higher in the plume than in surrounding groundwater, although ECA and per-bacterium levels of A(T) differed little in the plume and the surrounding uncontaminated groundwater. However, per-bacterium levels of DNA and A(T) varied six- and threefold, respectively, during a 6-h period of decreasing growth rate for an unidentified pseudomonad isolated from contaminated groundwater and grown in batch culture.(ABSTRACT TRUNCATED AT 250 WORDS)

[1]  D. Karl,et al.  Methodology and measurement of adenylate energy charge ratios in environmental samples , 1978 .

[2]  B. Riemann,et al.  Predictive value of adenylate energy charge for metabolic and growth states of planktonic communities in lakes , 1982 .

[3]  E. Thurman,et al.  Long-term fate of organic micropollutants in sewage-contaminated groundwater. , 1988, Environmental science & technology.

[4]  E. Thurman,et al.  Movement and fate of detergents in groundwater: a field study , 1986 .

[5]  D. LeBlanc.  Sewage plume in a sand and gravel aquifer , 1982 .

[6]  W. S. Silver.  Microbial ecology. , 1967, Science.

[7]  R. Harvey,et al.  Effect of organic contamination upon microbial distributions and heterotrophic uptake in a Cape Cod, Mass., aquifer , 1984, Applied and environmental microbiology.

[8]  R. Harvey,et al.  Growth determinations for unattached bacteria in a contaminated aquifer , 1987, Applied and environmental microbiology.

[9]  J. Romano,et al.  Ecological aspects of the surface microlayer. I. ATP, ADP, AMP contents, and energy charge ratios of microplanktonic communities , 1983 .

[10]  W. Sutcliffe,et al.  Difficulties with ATP measurements in inshore waters1 , 1976 .

[11]  J. Paul,et al.  Activity of an Attached and Free-Living Vibrio sp. as Measured by Thymidine Incorporation, p-Iodonitrotetrazolium Reduction, and ATP/DNA Ratios , 1986, Applied and environmental microbiology.

[12]  M. Coveney,et al.  Isocratic HPLC analysis of adenine nucleotides in environmental samples , 1986 .

[13]  N. Ireland,et al.  ADENYLATE ENERGY CHARGE MEASUREMENTS IN FRESHWATER MICROBIAL STUDIES , 1982 .

[14]  T. Bott,et al.  Adenylate energy charge in streambed sediments , 1985 .

[15]  D. J. Nuttley,et al.  Purification of DNA for bacterial productivity estimates , 1990, Applied and environmental microbiology.

[16]  Richard L. Smith,et al.  Importance of closely spaced vertical sampling in delineating chemical and microbiological gradients in groundwater studies , 1991 .

[17]  J. Wimpenny,et al.  Measurements of the distribution of adenylate concentrations and adenylate energy charge across Pseudomonas aeruginosa biofilms , 1992, Applied and environmental microbiology.

[18]  B. Olson,et al.  Fluorometric determination of the DNA concentration in municipal drinking water , 1985, Applied and environmental microbiology.

[19]  J. Hobbie,et al.  Use of nuclepore filters for counting bacteria by fluorescence microscopy , 1977, Applied and environmental microbiology.

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

[21]  Richard L. Smith,et al.  Denitrification in a Sand and Gravel Aquifer , 1988, Applied and environmental microbiology.

[22]  L. Barber,et al.  Associations of free-living bacteria and dissolved organic compounds in a plume of contaminated groundwater , 1992 .

[23]  J. Paul,et al.  Fluorometric determination of DNA in aquatic microorganisms by use of hoechst 33258. , 1982, Applied and environmental microbiology.

[24]  J. Paul,et al.  Genetic material in the marine environment: Implication for bacterial DNA , 1984 .

[25]  D. E. Atkinson,et al.  Adenylate Energy Charge in Escherichia coli During Growth and Starvation , 1971, Journal of bacteriology.

[26]  J. Fuhrman,et al.  Extraction from Natural Planktonic Microorganisms of DNA Suitable for Molecular Biological Studies , 1988, Applied and environmental microbiology.

[27]  D. White,et al.  Fluorometric Determination of Adenosine Nucleotide Derivatives as Measures of the Microfouling, Detrital, and Sedimentary Microbial Biomass and Physiological Status , 1980, Applied and environmental microbiology.

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