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TECHNICAL REPORT
Waste Management

Enzymatic Characterization of Organic Phosphorus in Animal Manure

USDA Agricultural Research Service, New England Plant, Soil, and Water Lab., Univ. of Maine, Orono, ME 04469

* Corresponding author (zhe{at}maine.edu)

Received for publication December 8, 2000. Information on the forms of P present in animal manure may improve our ability to manage manure P. In most investigations of manure P composition, only inorganic and total P are determined, and the difference between them is assigned as organic P. In this study, we explored the possibility of identifying and quantifying more specific organic P forms in animal manure with orthophosphate-releasing enzymes. Pig (Sus scrofa) manure and cattle (Bos taurus) manure were first sequentially fractionated into water-soluble P, NaHCO₃–soluble P, NaOH-soluble P, HCl-soluble P, and residual P. The fractions were separately incubated with wheat phytase, alkaline phosphatase, nuclease P1, nucleotide pyrophosphatase, or their combinations. The released orthophosphate was determined by a molybdate blue method. Part of the organic P in those fractions could be identified by the enzymatic treatments as phytate (i.e., 39% for pig manure and 17% for cattle manure in water-soluble organic P), simple phosphomonoesters (i.e., 43% for pig manure and 15% for cattle manure in NaOH-soluble organic P), nucleotide-like phosphodiesters (2–12%), and nucleotide pyrophosphate (0–4%). Our data indicate that the enzymatic treatment is an effective approach to identify and quantify the organic P forms present in animal manures.

Abbreviations: NMR, nuclear magnetic resonance • P_-a and P_+a, orthophosphate determined after samples were incubated in the absence and presence of alkaline phosphatase • P_-w and P_+w, orthophosphate determined after samples were incubated in the absence and presence of wheat phytase

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