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      Resolving the individual contribution of key microbial populations to enhanced biological phosphorus removal with Raman–FISH

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          Abstract

          Enhanced biological phosphorus removal (EBPR) is a globally important biotechnological process and relies on the massive accumulation of phosphate within special microorganisms. Candidatus Accumulibacter conform to the classical physiology model for polyphosphate accumulating organisms and are widely believed to be the most important player for the process in full-scale EBPR systems. However, it was impossible till now to quantify the contribution of specific microbial clades to EBPR. In this study, we have developed a new tool to directly link the identity of microbial cells to the absolute quantification of intracellular poly-P and other polymers under in situ conditions, and applied it to eight full-scale EBPR plants. Besides Ca. Accumulibacter, members of the genus Tetrasphaera were found to be important microbes for P accumulation, and in six plants they were the most important. As these Tetrasphaera cells did not exhibit the classical phenotype of poly-P accumulating microbes, our entire understanding of the microbiology of the EBPR process has to be revised. Furthermore, our new single-cell approach can now also be applied to quantify storage polymer dynamics in individual populations in situ in other ecosystems and might become a valuable tool for many environmental microbiologists.

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          Most cited references47

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          Recent advances in removing phosphorus from wastewater and its future use as fertilizer (1997-2003).

          Large quantities of phosphate present in wastewater is one of the main causes of eutrophication that negatively affects many natural water bodies, both fresh water and marine. It is desirable that water treatment facilities remove phosphorus from the wastewater before they are returned to the environment. Total removal or at least a significant reduction of phosphorus is obligatory, if not always fulfilled, in most countries. This comprehensive review summarizes the current status in phosphorus-removal technologies from the most common approaches, like metal precipitation, constructed wetland systems, adsorption by various microorganisms either in a free state or immobilized in polysaccharide gels, to enhanced biological phosphorus removal using activated sludge systems, and several innovative engineering solutions. As chemical precipitation renders the precipitates difficult, if not impossible, to recycle in an economical industrial manner, biological removal opens opportunities for recovering most of the phosphorus and beneficial applications of the product. This review includes the options of struvite (ammonium-magnesium-phosphate) and hydroxyapatite formation and other feasible options using, the now largely regarded contaminant, phosphorus in wastewater, as a raw material for the fertilizer industry. Besides updating our knowledge, this review critically evaluates the advantage and difficulties behind each treatment and indicates some of the most relevant open questions for future research.
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            Towards global phosphorus security: a systems framework for phosphorus recovery and reuse options.

            Human intervention in the global phosphorus cycle has mobilised nearly half a billion tonnes of the element from phosphate rock into the hydrosphere over the past half century. The resultant water pollution concerns have been the main driver for sustainable phosphorus use (including phosphorus recovery). However the emerging global challenge of phosphorus scarcity with serious implications for future food security, means phosphorus will also need to be recovered for productive reuse as a fertilizer in food production to replace increasingly scarce and more expensive phosphate rock. Through an integrated and systems framework, this paper examines the full spectrum of sustainable phosphorus recovery and reuse options (from small-scale low-cost to large-scale high-tech), facilitates integrated decision-making and identifies future opportunities and challenges for achieving global phosphorus security. Case studies are provided rather than focusing on a specific technology or process. There is no single solution to achieving a phosphorus-secure future: in addition to increasing phosphorus use efficiency, phosphorus will need to be recovered and reused from all current waste streams throughout the food production and consumption system (from human and animal excreta to food and crop wastes). There is a need for new sustainable policies, partnerships and strategic frameworks to develop renewable phosphorus fertilizer systems for farmers. Further research is also required to determine the most sustainable means in a given context for recovering phosphorus from waste streams and converting the final products into effective fertilizers, accounting for life cycle costs, resource and energy consumption, availability, farmer accessibility and pollution. Copyright © 2011 Elsevier Ltd. All rights reserved.
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              daime, a novel image analysis program for microbial ecology and biofilm research.

              Combinations of microscopy and molecular techniques to detect, identify and characterize microorganisms in environmental and medical samples are widely used in microbial ecology and biofilm research. The scope of these methods, which include fluorescence in situ hybridization (FISH) with rRNA-targeted probes, is extended by digital image analysis routines that extract from micrographs important quantitative data. Here we introduce daime (digital image analysis in microbial ecology), a new computer program integrating 2-D and 3-D image analysis and visualization functionality, which has previously not been available in a single open-source software package. For example, daime automatically finds 2-D and 3-D objects in images and confocal image stacks, and offers special functions for quantifying microbial populations and evaluating new FISH probes. A novel feature is the quantification of spatial localization patterns of microorganisms in complex samples like biofilms. In combination with '3D-FISH', which preserves the 3-D structure of samples, this stereological technique was applied in a proof of principle experiment on activated sludge and provided quantitative evidence that functionally linked ammonia and nitrite oxidizers cluster together in their habitat. This image analysis method complements recent molecular techniques for analysing structure-function relationships in microbial communities and will help to characterize symbiotic interactions among microorganisms.
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                Author and article information

                Contributors
                +45 9940 8503 , phn@bio.aau.dk
                Journal
                ISME J
                ISME J
                The ISME Journal
                Nature Publishing Group UK (London )
                1751-7362
                1751-7370
                20 March 2019
                20 March 2019
                August 2019
                : 13
                : 8
                : 1933-1946
                Affiliations
                [1 ]ISNI 0000 0001 0742 471X, GRID grid.5117.2, Department of Chemistry and Bioscience, Center for Microbial Communities, , Aalborg University, ; Aalborg, Denmark
                [2 ]ISNI 0000 0001 2286 1424, GRID grid.10420.37, Department of Microbial Ecology and Ecosystem Science, Research Network “Chemistry meets Microbiology”, , University of Vienna, ; Vienna, Austria
                [3 ]GRID grid.430357.6, Present Address: Department of Biological Sciences, Faculty of Applied Sciences, , Rajarata University, ; Mihintale, Sri Lanka
                Author information
                http://orcid.org/0000-0003-3749-8730
                http://orcid.org/0000-0002-9778-7684
                http://orcid.org/0000-0002-8747-6938
                http://orcid.org/0000-0002-6402-1877
                Article
                399
                10.1038/s41396-019-0399-7
                6776032
                30894691
                9b2bb076-aea2-4e3e-a0fc-2f00a6a31339
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 25 August 2018
                : 2 February 2019
                : 28 February 2019
                Funding
                Funded by: Innovation Fund Denmark, grant 4106-00014B and 1305-00018B
                Categories
                Article
                Custom metadata
                © International Society for Microbial Ecology 2019

                Microbiology & Virology
                water microbiology,soil microbiology
                Microbiology & Virology
                water microbiology, soil microbiology

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