The effect of key process operational conditions on enhanced biological phosphorus removal from wastewater

Enhanced biological phosphorus removal (EBPR) is the most economic and sustainable option used in wastewater treatment plants (WWTPs) for phosphorus removal. In this process it is important to control the competition between polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), since EBPR deterioration or failure can be related with the proliferation of GAOs over PAOs. This thesis is focused on the effect of operational conditions (volatile fatty acid (VFA) composition, dissolved oxygen (DO) concentration and organic carbon loading) on PAO and GAO metabolism. The knowledge about the effect of these operational conditions on EBPR metabolism is very important, since they represent key factors that impact WWTPs performance and sustainability. Substrate competition between the anaerobic uptake of acetate and propionate (the main VFAs present in WWTPs) was shown in this work to be a relevant factor affecting PAO metabolism, and a metabolic model was developed that successfully describes this effect. Interestingly, the aerobic metabolism of PAOs was not affected by different VFA compositions, since the aerobic kinetic parameters for phosphorus uptake, polyhydroxyalkanoates (PHAs) degradation and glycogen production were relatively independent of acetate or propionate concentration. This is very relevant for WWTPs, since it will simplify the calibration procedure for metabolic models, facilitating their use for full-scale systems. The DO concentration and aerobic hydraulic retention time (HRT) affected the PAO-GAO competition, where low DO levels or lower aerobic HRT was more favourable for PAOs than GAOs. Indeed, the oxygen affinity coefficient was significantly higher for GAOs than PAOs, showing that PAOs were far superior at scavenging for the often limited oxygen levels in WWTPs. The operation of WWTPs with low aeration is of high importance for full-scale systems, since it decreases the energetic costs and can potentially improve WWTP sustainability. Extended periods of low organic carbon load, which are the most common conditions that exist in full-scale WWTPs, also had an impact on PAO and GAO activity. GAOs exhibited a substantially higher biomass decay rate as compared to PAOs under these conditions, which revealed a higher survival capacity for PAOs, representing an advantage for PAOs in EBPR processes. This superior survival capacity of PAOs under conditions more closely resembling a full-scale environment was linked with their ability to maintain a residual level of PHA reserves for longer than GAOs, providing them with an effective energy source for aerobic maintenance processes. Overall, this work shows that each of these key operational conditions play an important role in the PAO-GAO competition and should be considered in WWTP models in order to improve EBPR processes

From food waste to volatile fatty acids towards a circular economy

The food industrial sector generates large amounts of waste, which are often used for animal feed, for agriculture or landfilled. However, these wastes have a very reach composition in carbon and other compounds, which make them very attractive for valorization through biotechnological processes. Added value compounds, such as volatile fatty acids (VFAs), can be produced by anaerobic fermentation using pure cultures or mixed microbial cultures and food waste as carbon source. Research on valuable applications for VFAs, such as polyhydroxyalkanoates, bioenergy or biological nutrient removal, towards a circular economy is emerging. This enhances the sustainability and the economic value of food waste. This chapter reviews the various types of food waste used for VFAs production using mixed microbial cultures, the anaerobic processes, involved and the main applications for the produced VFAs. The main parameters affecting VFAs production are also discussed.publishersversionpublishe

Multimédia interativo como suporte à comunicação em saúde

Mestrado em Comunicação MultimédiaA presente investigação visa compreender como é possível através de uma aplicação multimédia influenciar a comunicação em saúde. Neste sentido, pretende-se desenvolver uma proposta de estratégia de comunicação para crianças com diabetes tipo 1. Com o objetivo de comprovar as hipóteses traçadas foram entrevistadas crianças com idades compreendidas entre os 6 e os 10 anos portadoras de diabetes tipo 1 acompanhadas no Hospital Santa Maria em Lisboa, assim como a equipa de profissionais de saúde e os pais das crianças que participaram na investigação, onde se veio a apurar a sua visão sobre a utilidade de uma aplicação multimédia na compreensão inicial da diabetes. Os dados recolhidos revelam que a existência de uma aplicação multimédia que pudesse ensinar às crianças os principais cuidados a ter com a diabetes assim como lhes explicasse que procedimentos a adotar em determinadas situações poderia facilitar a compreensão inicial da diabetes, assim como facilitaria a mudança de hábitos, tanto para as crianças como para os pais.This research aims to understand how it is possible via a multimedia application to influence health communication. In this sense, we intend to develop a proposal for a communication strategy for children with type 1 diabetes. Aiming to prove the hypotheses outlined we interviewed children aged between 6 and 10 years old with type 1 diabetes followed at the Hospital Santa Maria in Lisbon, as well as the team of health professionals and parents, where we could establish the view on the usefulness of a multimedia application in initial understanding of diabetes. The data collected show that the existence of a multimedia application that could teach children the primary care of diabetes as it explains what procedures to follow in certain situations could facilitate the initial understanding of diabetes, as well as facilitate the changing of habits both for children and for parents

From Food Waste to Volatile Fatty Acids towards a Circular Economy

The food industrial sector generates large amounts of waste, which are often used for animal feed, for agriculture or landfilled. However, these wastes have a very reach composition in carbon and other compounds, which make them very attractive for valorization through biotechnological processes. Added value compounds, such as volatile fatty acids (VFAs), can be produced by anaerobic fermentation using pure cultures or mixed microbial cultures and food waste as carbon source. Research on valuable applications for VFAs, such as polyhydroxyalkanoates, bioenergy or biological nutrient removal, towards a circular economy is emerging. This enhances the sustainability and the economic value of food waste. This chapter reviews the various types of food waste used for VFAs production using mixed microbial cultures, the anaerobic processes, involved and the main applications for the produced VFAs. The main parameters affecting VFAs production are also discussed

Intracellular poly-P assessment by DAPI staining and image analysis

In wastewater treatment, enhanced biological phosphorus removal (EBPR) is considered a well-established process to remove phosphate (P). EBPR is based on the activity of polyphosphate-accumulating organisms (PAOs) able to take up and store large amounts of P as intracellular (poly-P) granules. However, monitoring poly-P in mixed cultures is usually performed by a laborious and time consuming off-line chemical analysis. Thus, there is a clear need to develop new techniques to rapidly monitor these processes, such as image analysis coupled to sample staining and microscopy inspection. A lab-scale sequencing batch reactor (SBR) was fed with synthetic wastewater containing acetate and propionate as main carbon sources and an orthophosphate solution was added. A COD/P ratio of 10 mg COD mg P-PO4-1 was used to provide selective advantages to PAOs. The SBR was operated with a cycle time of 6 h: 120 min anaerobic including 5 min feed, 180 min aerobic and 60 min wasting/settling. Biomass samples were collected at the end of the aerobic stage. Bulk P concentration was determined by segmented flow analysis and total P concentration was similarly measured following acid digestion at 100oC. Intracellular poly-P concentration was determined by subtracting the bulk P from the total P. Intracellular poly-P granules were observed in epifluorescence microscopy using DAPI staining with a 25 ìg mL-1 DAPI solution. A long pass filter was used with an excitation bandpass of 365-370 nm and emission cut off at 421 nm. A specially developed program in Matlab was used for image analysis. A total of 41 samples were collected. Two thirds were fed as training data to the partial least squares (PLS) model and the remaining used for validation. Both absolute (in mg poly-P / L) and relative (in mg poly-P / g MLSS) intracellular poly-P concentrations were studied. This procedure was found to predict, at some extent, the relative intracellular poly-P concentration (real poly-P = 0.971 x predicted poly-P, R2 of 0.744). Regarding the absolute intracellular poly-P concentration, a total of 3 samples needed to be discarded in order to obtain a similar result (real poly-P = 1.005 x predicted poly-P, R2 of 0.731)

Pilot-scale valorisation of salmon peptone into polyhydroxyalkanoates by mixed microbial cultures under conditions of high ammonia concentration

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Donnan Dialysis for Recovering Ammonium from Fermentation Solutions Rich in Volatile Fatty Acids

Funding Information: Universitat Politècnica de València and Ministerio de Universidades de España (Plan de Recuperación, Transformación y Resiliencia—financed by European Union—Next GenerationEU) are acknowledged for the post-doctoral research grant attributed to Kayo Santana Barros. Funding Information: This work was financed by Fundação para a Ciência e a Tecnologia, I.P., Lisbon, Portugal in the scope of the projects UIDP/04378/2020, UIDB/04378/2020 and PTDC/BTA-BTA/30902/2017 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. The work was also supported through the projects UIDB/50006/2020 and UIDP/50006/2020, funded by FCT/MCTES through national funds. Publisher Copyright: © 2023 by the authors.For the production of polyhydroxyalkanoates (PHA) using nitrogen-rich feedstocks (e.g., protein-rich resources), the typical strategy of restricting cell growth as a means to enhance overall PHA productivity by nitrogen limitation is not applicable. In this case, a possible alternative to remove the nitrogen excess (NH4+/NH3) is by applying membrane separation processes. In the present study, the use of Donnan dialysis to separate ammonium ions from volatile fatty acids present in the media for the production of PHA was evaluated. Synthetic and real feed solutions were used, applying NaCl and HCl receiver solutions separated by commercial cation-exchange membranes. For this specific purpose, Fumasep and Ralex membranes showed better performance than Ionsep. Sorption of ammonium ions occurred in the Ralex membrane, thus intensifying the ammonium extraction. The separation performances with NaCl and HCl as receiver solutions were similar, despite sorption occurring in the Ralex membrane more intensely in the presence of NaCl. Higher volumetric flow rates, NaCl receiver concentrations, and volume ratios of feed:receiver solutions enhanced the degree of ammonium recovery. The application of an external electric potential difference to the two-compartment system did not significantly enhance the rate of ammonium appearance in the receiver solution. The results obtained using a real ammonium-containing solution after fermentation of cheese whey showed that Donnan dialysis can be successfully applied for ammonium recovery from such solutions.publishersversionpublishe

Novel method to quantify intracellular accumulation of polyphosphate in EBPR systems

A new method for intracellular storage polyphosphate (poly-P)identification and quantification in enhanced biological phosphorus removal (EBPR) systems is proposed based on image analysis. In EBPR systems, 4',6-diamidino-2-phenylindole (DAPI) is usually combined with fluororescent in situ hybridization (FISH) to evaluate the microbial community. The proposed technique is based on an image analysis procedure specifically developed for determining poly-P inclusions within biomass suspension using solely DAPI by epifluorescence microscopy. Due to contradictory literature DAPI concentrations used for poly-P detection, the present work assessed the optimal DAPI concentration for samples acquired at the end of the EBPR aerobic stage when the accumulation is performed. Digital images were then acquired and processed by means of image processing and analysis. Regarding image analysis results and considering the current operational conditions, a promising correlation could be found between average poly-P intensity values and the analytical determination, although presenting a correlation coefficient somewhat far from the ideal. The proposed methodology can be seen as a promising alternative procedure to quantify intracellular poly-P accumulation in a faster and less labor intensive way

Biopolymer monitoring using quantitative image analysis techniques

Polyhydroxyalkanoates (PHAs) are intracellular biopolymers with many applications, particularly as substitutes of polypropylene and polyethylene, due to their thermoplastic properties and biocompatible nature. Furthermore, glycogen is a polysaccharide of glucose with high importance in the metabolism of microbial communities and polyphosphate is a microbial storage compound that should be recovered in order to offset the worldwide depletion of phosphorus sources. The determination of these biopolymers by chemical analysis is a laborious task, often involving digestion processes prior to gas and high-performance liquid chromatography, which are time consuming and difficult to apply in industry. Currently, it is important to develop new, rapid and simple techniques to monitor these polymers. Image analysis is a non-invasive and rapid technique that has the potential to be used to quantify these intracellular polymers quickly, in real-time. Mesquita et al. (2013) showed that it is possible to predict the concentration of glycogen and PHAs by quantitative image analysis, using aniline blue and nile blue staining, respectively. Polyphosphate can also be predicted by this technique through DAPI staining, which is currently under development. These biopolymers are produced by several different microorganisms, and combining their quantification with fluorescence in situ hybridization (FISH) techniques for microbial identification can enable the determination of organisms that store high quantities of each biopolymer. In this work, an advanced quantitative technique is developed to perform real time monitoring of these three biopolymers in a bioreactor performing biological phosphorus removal. Image analysis of the biopolymers was combined with FISH to determine the storage level of each compound within the different microbial populations. This technique will further enable the assessment of biopolymer levels within microbial communities, which can be applied in the biopolymer production industry

Valorization of brewery waste through Polyhydroxyalkanoates production supported by a metabolic specialized microbiome

Raw brewers’ spent grain (BSG), a by-product of beer production and produced at a large scale, presents a composition that has been shown to have potential as feedstock for several biological processes, such as polyhydroxyalkanoates (PHAs) production. Although the high interest in the PHA production from waste, the bioconversion of BSG into PHA using microbial mixed cultures (MMC) has not yet been explored. This study explored the feasibility to produce PHA from BSG through the enrichment of a mixed microbial culture in PHA-storing organisms. The increase in organic loading rate (OLR) was shown to have only a slight influence on the process performance, although a high selectivity in PHA-storing microorganisms accumulation was reached. The culture was enriched on various PHA-storing microorganisms, such as bacteria belonging to the Meganema, Carnobacterium, Leucobacter, and Paracocccus genera. The enrichment process led to specialization of the microbiome, but the high diversity in PHA-storing microorganisms could have contributed to the process stability and efficiency, allowing for achieving a maximum PHA content of 35.2 ± 5.5 wt.% (VSS basis) and a yield of 0.61 ± 0.09 CmmolPHA/CmmolVFA in the accumulation assays. Overall, the production of PHA from fermented BSG is a feasible process confirming the valorization potential of the feedstock through the production of added-value products.info:eu-repo/semantics/publishedVersio