Critical problems with extraction of ATP for bioluminescence assay of plankton biomass |
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Authors: | William B Perry James T Boswell Jack A Stanford |
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Institution: | (1) Department of Biological Sciences, North Texas State University, 76203, 817/788-2011 Denton, Texas;(2) Texas Instruments, Inc., Ecological Services Group, P.O. Box 5621, 75222 Dallas, Texas |
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Abstract: | Recovery of ATP by boiling tris extraction was 90–95 percent greater in 1 liter grab samples than in concentrated net samples. ATP losses were attributed to insulating effects promoted by accumulation of detritus on filters. A series of extractions over a concentration range of whole or size-segregated plankters and cultured algae was made to determine volume of water to be filtered for optimum extraction efficiency. Accuracy of ATP assays was optimized by: (i) using large diameter (i.e. 47 mm) acetate filters; (2) limiting sample volume filtered to 50 ml when particulate organic carbon (POC) exceeded 0.4 mg l–1; and (3) performing extractions in boiling tris maintained initially on a laboratory hot plate at 400°C as opposed to hot water bath at 100°C.Additional problems were encountered in using published cellular carbon: ATP ratios for conversion of ATP data to biomass as carbon. Ratios of POC: ATP in cultures of sheathed blue-green algae reached 550 : i, while non-sheathed forms yielded ratios near values previously reported for plankton communities. Difficulties in applying a uniform conversion factor may be expected in plankton communities containing significant volumes of sheathed blue-greens. |
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Keywords: | ATP microbial biomass limnological methods plankton |
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