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Improved methodology for ATP determination in marine environments

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Abstract

A simple technique is described for de-salting and concentrating adenosine triphosphate (ATP) extracted from seawater or marine sediment samples prior to assays with the standard luciferin-luciferase procedure. The technique involves chromatography of H2SO4 extracts on columns of activated carbon. The efficiency of ATP recovery from marine sediments using this pre-treatment was superior to that attained with either boiling “Tris” extraction or with H2SO4 extraction without subsequent purification. All ions which interfere with the luciferase reaction or precipitate ATP upon neutralization of acid extracts are removed with this procedure, thus eliminating the 50- to 100-fold dilutions required with other acid-extraction procedures. In addition, the purified ATP extracts may be concentrated up to 100-fold, thus greatly improving the sensitivity of ATP measurement in samples containing very low biomass. This procedure has been applied to ATP determination in marine sediments in the upwelling area off northwest Africa. The average ATP concentrations in the upper 6 cm of sediment from 1000 and 200 m were 195 and 545 ng per ml wet sediment, respectively.

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Literature Cited

  • Atkinson, D.E.: Regulation of enzyme function. A. Rev. Microbiol. 23, 47–68 (1969)

    Google Scholar 

  • Ausmus, B.S.: The use of the ATP assay in terrestrial decomposition studies. In: Modern methods in the study of microbial ecology, pp 223–234. Ed. by T. Rosswall. Stockholm: Swedish Natural Science Research Council (NRF) 1972

    Google Scholar 

  • Birkett, L. and A.D. McIntyre: Treatment and sorting of samples. In: IBP Handbook No. 16. Methods for the study of marine benthos, pp 156–168. Ed. by N.A. Holme and A.D. McIntyre. Oxford: Blackwell Scientific Publications 1971

    Google Scholar 

  • Carlucci, A.F. and D. Pramer: Factors influencing the plate method for determining abundance of bacteria in sea water. Proc. Soc. exp. Biol. Med. 96, 392–394 (1957)

    Google Scholar 

  • Christian, R.R., K. Bancroft and W.J. Wiebe: Distribution of microbial adenosine triphosphate in salt marsh sediments at Sapelo Island, Georgia. Soil Sci. 119, 89–97 (1975)

    Google Scholar 

  • Ehrlich, H.: Bacteriology of manganese nodules. I. Bacterial action on manganese nodule enrichments. Appl. Microbiol. 11, 15–19 (1963)

    Google Scholar 

  • Ernst, W.: ATP als Indikator für die Biomasse mariner Sedimente. Oecologia (Berl.) 5, 55–60 (1970)

    Google Scholar 

  • Ferguson, R.L. and M.B. Murdoch: On the estimation of microbial ATP in estuarine sediments and water. Bull. ecol. Soc. Am. 54, p. 27 (1973)

    Google Scholar 

  • Griggs, G.B., A.G. Carey and L.D. Kulm: Deep-sea sedimentation and sediment-fauna interaction in Cascadia Channel and on Cascadia Abyssal Plain. Deep-Sea Res. 16, 157–170 (1969)

    Google Scholar 

  • Holm-Hansen, O.: ATP levels in algal cells as influenced by environmental conditions. Pl. Cell Physiol., Tokyo 11, 689–700 (1970)

    Google Scholar 

  • — Determination of total microbial biomass by measurement of adenosine triphosphate. In: Estuarine microbial ecology, pp 73–89. Ed. by L.H. Stevenson and R.R. Colwell. Columbia, S.C.: University of South Carolina Press 1973

    Google Scholar 

  • — and C.R. Booth: The measurement of adenosine triphosphate in the ocean and its ecological significance. Limnol. Oceanogr. 11, 510–519 (1966)

    Google Scholar 

  • — and H.W. Paerl: The applicability of ATP determination for estimation of microbial biomass and metabolic activity. In: Detritus and its ecological role in aquatic ecosystems, pp 149–168. Ed. by U. Melchiorri. Pallanza: Istituto Italiano di Idrobiologia 1972

    Google Scholar 

  • Ireland, D.M. and D.C.B. Mills. Detection and determination of adenosine diphosphate and related substances in plasma. Biochem. J. 99, 283–296 (1966)

    Google Scholar 

  • Jannasch, H., K. Eimhjellen, C. Wirson and A. Farmanfarmaian: Microbial degradation of organic matter in the deep sea. Science, N.Y. 171, 672–675 (1971)

    Google Scholar 

  • Johnson, R.W. and J. Calder: Early diagenesis of fatty acids and hydrocarbon in a salt marsh environment. Geochim. cosmochim. Acta 37, 1943–1955 (1973)

    Google Scholar 

  • Karl, D.M. and P.A. LaRock: Adenosine triphosphate measurements in soil and marine sediments. J. Fish. Res. Bd Can. 32, 599–607 (1975)

    Google Scholar 

  • Karl, D.M. P.A. LaRock, J.W. Morse and W. Sturges: Adenosine triphosphate in the North Atlantic Ocean and its relationship to the oxygen minimum. Deep-Sea Res. (1975). (In press)

  • Lee, C.C., R.F. Harris, J.D.H. Williams, D.E. Armstrong and J.K. Syers: ATP in lake sediments; determination. Proc. Soil Sci. Soc. Am. 35, 82–86 (1971a)

    Google Scholar 

  • ——, J.K. Syers and D.E. Armstrong: ATP in lake sediments: II-origin and significance. Proc. Soil Sci. Soc. Am. 35, 86–91 (1971b)

    Google Scholar 

  • Litchfield, C.: Interactions of amino acids and marine bacteria. In: Estuarine microbial ecology, pp 145–168. Ed. by L.H. Stevenson and R.R. Colwell. Columbia, S.C.: University of South Carolina Press 1973

    Google Scholar 

  • Menzies, R.J. and G.T. Rowe: The LUBS, a large undisturbed bottom sampler. Limnol. Oceanogr. 13, 708–714 (1968)

    Google Scholar 

  • Patterson, J.W., P.L. Brezonik and H.D. Putman: Measurement and significance of ATP in activated sludge. Envir. Sci. Technol. 4, 569–575 (1970)

    Google Scholar 

  • Sugahara, I., M. Sugiyama and A. Kawai: Distribution and activity of nitrogen-cycle bacteria in water-sediment systems with different concentrations of oxygen. In: Effect of the ocean environment on microbial activities, pp 327–340. Ed. by R.R. Colwell and R. Morita. Baltimore: University Park Press 1974

    Google Scholar 

  • Temple, K.: Syngenesis of sulfide ores: an evaluation of biochemical aspects. Econ. Geol. 59, 1473–1491 (1964)

    Google Scholar 

  • Weddle, C.L. and D. Jenkins: The viability and activity of activated sludge. Wat. Res. 5, 621–640 (1971)

    Google Scholar 

  • Wright, R.: Mineralization of organic solutes by heterotrophic bacteria. In: Effect of the ocean environment on microbial activities, pp 546–565. Ed. by R.R. Colwell and R. Morita. Baltimore: University Park Press 1974

    Google Scholar 

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Communicated by J.S. Pearse, Santa Cruz

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Hodson, R.E., Holm-Hansen, O. & Azam, F. Improved methodology for ATP determination in marine environments. Marine Biology 34, 143–149 (1976). https://doi.org/10.1007/BF00390756

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