Methods for Detection and Quantification of Polyphosphate and Polyphosphate Accumulating Microorganisms in Aquatic Sediments

Abstract

It has been speculated that the microbial P pool is highly variable in the uppermost layer of various aquatic sediments, especially when an excessive P accumulation in form of polyphosphate (Poly-P) occurs. Poly-P storage is a universal feature of many different organisms and has been technically optimised in wastewater treatment plants (WWTP) with enhanced biological phosphorus removal (EBPR). In the recent past, new insights into mechanisms of P elimination in WWTP almost exclusively depended on the development and application of novel methods like 31 P-NMR spectroscopy and molecular methods for identifying Poly-P accumulating microorganisms (PAO). The aim of the present review is to compile current methods potentially available for detection and quantification of Poly-P in sediments and to complement it with yet unpublished results to validate their application in natural sediments. The most powerful tool for reliable Poly-P quantification in sediments is the liquid 31 P-NMR technique which has been successfully used for Poly-P measurements in a variety of aquatic sediments. But the microorganisms as well as mechanisms involved in Poly-P storage and cycling are largely unknown. Therefore, we also intend to stimulate future studies focusing on these encouraging topics in sediment research via the implementation of novel methods.

Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

“…Phosphorus-31 nuclear magnetic resonance spectroscopy requires large sample sizes and may be insensitive to polyP tightly associated with metal cations in solid granules. Moreover, alkaline extractions may incompletely extract and partly hydrolyze polyP (30,31). Assays based on enzymes of polyP metabolism (either exopolyphosphatase followed by orthophosphate [PO 4 3Ϫ ] determination [32,33] or polyP kinase followed by ATP measurement [34,35]) entail purifying polyP, but purification procedures suffer from low yields (36,37) and there is no commercial source for the necessary enzymes.…”

Seasonal and spatial variability of sediment bacterial communities inhabiting the large shallow Lake Balaton

“…It is more effective than other staining procedures, because it produces higher contrast between the cell and the granules (Serafim et al, 2002). However, because of the detection limits of the optical microscopy, and knowing that none of the available dyes are exclusively specific for poly-P, staining results must be judged with caution and Neisser positive bacteria have to be indicated as potential PAO until storage material can be chemically characterized as poly-P (Kulaev & Kulakovskaya, 2000;Hupfer et al, 2008).…”

Microbial activity facilitates phosphorus adsorption to shallow lake sediment

“…When P levels were adequate in the water, a substantial portion of P was stored in microbial cells in the form of nucleic acids, phospholipids, sugar phosphate, and other organic P compounds (Haglund et al 2003;Uhlmann et al 1998). Some of these compounds may be assimilated by the decomposers and some may be released as inorganic P. It has been suggested that some of the fixed P may eventually become particulate refractory organic P compounds and was buried in the sediments (Gächter and Meyer 1993;Hupfer et al 2008). The microbial P adsorption is a process during which microorganisms assimilate P for development.…”