Wanneer we een diepere kijk nemen in de compositie van het afvalslib, zien we dat het bestaat uit een heleboel verschillende componenten. Een van deze componenten zijn de extracellulaire polymeren, ook wel EPS genaamd. Dit zijn grote biologische moleculen, denk bijvoorbeeld aan suikers en eiwitten, die potentieel hebben om gebruikt te worden in een hele resem aan toepassingen. De mogelijkheden zijn enorm breed, gaande van gebruik als coating om materialen meer waterafstotend of brandwerend te maken, alsook als mogelijke vervanger van de commerciële, olie-gebaseerde plastics. Een derde toepassing, waarop deze thesis heeft gefocust, is het gebruik van EPS als hydrogel, een quasi vaste stof die voornamelijk bestaat uit water. Deze gels kunnen gebruikt worden voor de verwijdering van zware metalen uit de bodem, of als biologisch afbreekbare meststof, die ook nog eens veel gerichter tewerk gaat dan standaard bemesting. Deze EPS zijn dus duidelijk een potentiële drijfveer om het hergebruik van afvalwaterslib in gang te zetten.
Om binnen een industriële toepassing bruikbaar te zijn, is het belangrijk dat EPS uit verschillende bronnen een zo vast mogelijke compositie hebben. Wanneer dit het geval is, kan je deze verschillende EPS’en gebruiken voor eenzelfde toepassing. Dit zorgt voor een grotere hoeveelheid geschikte grondstoffen, wat je proces makkelijker opschaalbaar maakt. Om na te gaan of EPS van verschillende bronnen ook een andere samenstelling hebben, zijn verscheidene chemische karakterisatietechnieken toegepast op EPS van verschillende bronnen. Zowel eenvoudige pure culturen, bestaande uit één type bacteriën, alsook verschillende industriële afvalslibben, die vele verschillende bacteriën bevatten, zijn bestudeerd. Er werd ook onderscheid gemaakt tussen zoutwater- en zoetwatersystemen. Deze karakterisatie toonde aan dat EPS van verschillende bronnen een unieke samenstelling hebben. Dit maakt het hergebruik van het afvalslib aanzienlijk moeilijker. Het is namelijk mogelijk dat de variabele EPS compositie leidt tot variabele eigenschappen van het eindproduct, in dit geval de hydrogel. Idealiter zouden EPS van alle verschillende origines resulteren in hydrogels met gelijke of zeer gelijkaardige eigenschappen. Om deze reden zijn de EPS afkomstig van de industriële slibben gebruikt om ook effectief hydrogels te maken, om zo het effect van de variabele compositie op de eigenschappen van de hydrogels na te gaan.
Met behulp van een zelfgemaakte 3D-geprinte mal zijn vier verschillende EPS hydrogels gemaakt. De stevigheid van deze gels is gemeten met behulp van een reometer, die bepaalt hoe makkelijk de hydrogels vervormen, wanneer je er een kracht op uitoefent. Hoe minder vervormbaar, hoe steviger de gel.

Afhankelijk van de toepassing is dit al dan niet gewenst. Denk bijvoorbeeld aan een matras: je wilt niet door je matras zakken, omdat ze te zacht (of niet stevig) is. Aan de andere kant slaap je ook niet graag op een te stevige matras, die aanvoelt als een baksteen. Het belang van de stevigheid van je hydrogel is dus niet te onderschatten. Uit enkele metingen bleek dat de samenstelling van de EPS een aanzienlijk effect had op de stevigheid van de hydrogel. Met deze reden is een poging gedaan om een verband te vinden tussen de compositie van de EPS en de stevigheid van diens hydrogel. Dit was helaas niet mogelijk, doordat de EPS een mengsel zijn van talloze en tot nog toe onbekende suikers en eiwitten. Het vinden van zo'n relatie was bijgevolg te complex.

Naast de moeilijkheidsgraad, veroorzaakt door de variabiliteit van de EPS compositie, is er nog een bijkomende moeilijkheid om ze commercieel te gaan gebruiken. EPS maken namelijk slechts één onderdeel uit van het afvalslib. Om deze reden is een extractieprotocol nodig, voordat ze gebruikt kunnen worden. Idealiter resulteert zo’n extractieprotocol in een fractie die EPS bevat en een EPS-loze pellet, die al het overige materiaal bevat. Ook dit blijkt makkelijker gezegd dan gedaan, aangezien verschillende extractieprotocols resulteren in verschillende EPS composities. Dit werd nagegaan door vier verschillende protocols met elkaar te vergelijken. Hieruit bleek dat de samenstelling van de geëxtraheerde EPS sterk afhankelijk is van het toegepaste protocol. Zelfs extracties die gebruik maken van dezelfde chemische stoffen, maar andere parameters (zoals tijd, temperatuur etc.) resulteren in een andere EPS samenstelling. Het is dus ook in dit geval belangrijk om het extractieprotocol te standaardiseren, om op deze manier zo weinig mogelijk variabiliteit te verkrijgen in de EPS compositie.
Het is dus duidelijk dat het hergebruik van afvalslib nog voor enkele grote uitdagingen staat. Een belangrijke rol in dit proces gaat naar de extracellulaire polymeren, ook wel EPS genaamd. Deze hebben enerzijds veel potentieel om gebruikt te worden in verscheidene toepassingen, anderzijds brengen deze ook enkele moeilijkheden met zich mee, zoals de afhankelijkheid van de EPS compositie op het type slib dat gebruikt is. Ook het gebrek aan een gestandaardiseerd extractieprotocol maakt dit niet makkelijker. Deze variabiliteiten hebben duidelijke effecten op het gebruik van EPS in toepassingen zoals hydrogels. Mijn studie toonde aan dat deze toepassing mogelijk is.
C. Abeygunawardana, T. C. Williams, J. S. Sumner, and J. P. Hennessey. Development and Validation of an NMR-Based Identity Assay for Bacterial Polysaccharides. Analytical Biochemistry, 279(2):226–240, Mar. 2000. DOI: 10.1006/abio.1999.4470.
[2] S. S. Adav and D.-J. Lee. Extraction of extracellular polymeric substances from aerobic granule with compact interior structure. Journal of Hazardous Materials, 154(1-3):1120–1126, June 2008. DOI: 10.1016/j.jhazmat.2007.11.058.
[3] S. S. Adav, D.-J. Lee, and J.-H. Tay. Extracellular polymeric substances and structural stability of aerobic granule. Water Research, 42(6-7):1644–1650, Mar. 2008. DOI: 10.1016/j.watres.2007.10.013.
[4] T. Agustina, H. Ang, and V. Vareek. A review of synergistic effect of photocatalysis and ozonation on wastewater treatment. Journal of Photochemistry and Photobiology C: Photochemistry Reviews, 6(4):264–273, Dec. 2005. DOI: 10.1016/j.jphotochemrev.2005.12.003.
[5] K. M. Alkaabi, A. Yafea, and S. S. Ashraf. Effect of pH on Thermal- and Chemical-Induced Denaturation of GFP. Applied Biochemistry and Biotechnology, 126(2):149–156, 2005. DOI: 10.1385/abab:126:2:149.
[6] American Public Health Association (APHA). Standard Methods for the Examination of Water and Wastewater. American Public Health Association, Washington, D.C., 20th edition, 2012.
[7] M. Arienzo, E. Christen, W. Quayle, and A. Kumar. A review of the fate of potassium in the soil-plant system after land application of wastewaters. Journal of Hazardous Materials, 164(2-3):415–422, May 2009. DOI: 10.1016/ j.jhazmat.2008.08.095.
[8] N. R. Babij, E. O. McCusker, G. T. Whiteker, B. Canturk, N. Choy, L. C. Creemer, C. V. D. Amicis, N. M. Hewlett, P. L. Johnson, J. A. Knobelsdorf, F. Li, B. A. Lorsbach, B. M. Nugent, S. J. Ryan, M. R. Smith, and Q. Yang. NMR Chemical Shifts of Trace Impurities: Industrially Preferred Solvents Used in Process and Green Chemistry. Organic Process Research & Development, 20(3):661–667, Feb. 2016. DOI: 10.1021/acs.oprd.5b00417.
[9] A. S. Bal and N. N. Dhagat. Upflow anaerobic sludge blanket reactor–a review. Indian journal of environmental health, 43:1–82, Apr. 2001. [10] B. Balasubramanian, S. Ilavenil, A.-D. N.A., P. Agastian, and K. C. Choi. Isolation and characterization of Aspergillus sp. for the production of extracellular polysaccharides by response surface methodology. Saudi Journal of Biological Sciences, 26(3):449–454, Mar. 2019. DOI: 10.1016/j.sjbs.2018.10.015. [11] A. Bateman. The structure of a domain common to archaebacteria and the homocystinuria disease protein. Trends in Biochemical Sciences, 22(1):12–13, Jan. 1997. DOI: 10.1016/s0968-0004(96)30046-7. [12] R. Benedict and D. Carlson. Aerobic heterotrophic bacteria in activated sludge. Water Research, 5(11):1023–1030, Nov. 1971. DOI: 10.1016/0043-1354(71) 90036-4. [13] M. Boleij, M. Pabst, T. R. Neu, M. C. M. van Loosdrecht, and Y. Lin. Identification of Glycoproteins Isolated from Extracellular Polymeric Substances of Full-Scale Anammox Granular Sludge. Environmental Science and Technology, 52(22):13127–13135, Oct. 2018. DOI: 10.1021/acs.est.8b03180. [14] M. Boleij, T. Seviour, L. L. Wong, M. C. van Loosdrecht, and Y. Lin. Solubilization and characterization of extracellular proteins from anammox granular sludge. Water Research, 164:114952, Nov. 2019. DOI: 10.1016/j.watres. 2019.114952.
[15] S. Bolisetty, M. Peydayesh, and R. Mezzenga. Sustainable technologies for water purification from heavy metals: review and analysis. Chemical Society Reviews, 48(2):463–487, 2019. DOI: 10.1039/c8cs00493e.
[16] U. Bose, J. A. Broadbent, A. Juhasz, S. Karnaneedi, E. B. Johnston, S. Stockwell, K. Byrne, V. Limviphuvadh, S. Maurer-Stroh, A. L. Lopata, and M. L. Colgrave. Protein extraction protocols for optimal proteome measurement and arginine kinase quantitation from cricket acheta domesticus for food safety assessment. Food Chemistry, 348:129110, June 2021. DOI: 10.1016/j.foodchem.2021.129110.
[17] A. Bou-Sarkis, B. Pagliaccia, A. Ric, N. Derlon, E. Paul, Y. Bessiere, and E. Girbal-Neuhauser. Effects of alkaline solvents and heating temperatures on the solubilization and degradation of gel-forming Extracellular Polymeric Substances (EPS) extracted from aerobic granular sludge. Biochemical Engineering Journal, 185:108500, July 2022. DOI: 10.1016/j.bej.2022.108500.
[18] E. Brauchle, A. Knopf, H. Bauer, N. Shen, S. Linder, M. Monaghan, K. Ellwanger, S. Layland, S. Brucker, A. Nsair, and K. Schenke-Layland. Noninvasive Chamber-Specific Identification of Cardiomyocytes in Differentiating Pluripotent Stem Cells. Stem Cell Reports, 6(2):188–199, Feb. 2016. DOI: 10.1016/j.stemcr.2015.12.007.
[19] R. B. Brown and J. Audet. Current techniques for single-cell lysis. Journal of The Royal Society Interface, 5(2), Apr. 2008. DOI: 10.1098/rsif.2008.0009. focus. [20] C. Buenaño Vargas, M. C. Gagliano, L. M. Paulo, A. Bartle, A. Graham, H. P. J. van Veelen, and V. O’Flaherty. Acclimation of microbial communities to low and moderate salinities in anaerobic digestion. Science of The Total Environment, 906:167470, Jan. 2024. DOI: 10.1016/j.scitotenv.2023.167470. [21] G. S. Bumbrah and R. M. Sharma. Raman spectroscopy - Basic principle, instrumentation and selected applications for the characterization of drugs of abuse. Egyptian Journal of Forensic Sciences, 6(3):209–215, Sept. 2016. DOI: 10.1016/j.ejfs.2015.06.001. [22] J. T. Bunce, E. Ndam, I. D. Ofiteru, A. Moore, and D. W. Graham. A Review of Phosphorus Removal Technologies and Their Applicability to Small-Scale Domestic Wastewater Treatment Systems. Frontiers in Environmental Science, 6, Feb. 2018. DOI: 10.3389/fenvs.2018.00008. [23] K. D. Burch, B. Han, J. Pichtel, and T. Zubkov. Removal efficiency of commonly prescribed antibiotics via tertiary wastewater treatment. Environmental Science and Pollution Research, 26(7):6301–6310, Jan. 2019. DOI: 10.1007/s11356-019-04170-w. [24] K. Burton. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. The Biochemical journal, 62:315–323, Feb. 1956. [25] R. Campo, E. Carretti, C. Lubello, and T. Lotti. Recovery of structural extracellular polymeric substances (sEPS) from aerobic granular sludge: Insights on biopolymers characterization and hydrogel properties for potential applications. Journal of Environmental Management, 324:116247, Dec. 2022. DOI: 10.1016/j.jenvman.2022.116247. [26] V. L. Campo, D. F. Kawano, D. B. d. Silva, and I. Carvalho. Carrageenans: Biological properties, chemical modifications and structural analysis - A review. Carbohydrate Polymers, 77(2):167–180, June 2009. DOI: 10.1016/j.carbpol. 2009.01.020. [27] F. Cao, I. Bourven, P. N. Lens, E. D. van Hullebusch, Y. Pechaud, and G. Guibaud. Hydrophobic features of EPS extracted from anaerobic granular sludge: an investigation based on DAX-8 resin fractionation and size exclusion chromatography. Applied Microbiology and Biotechnology, 101(8):3427–3438, Dec. 2016. DOI: 10.1007/s00253-016-8053-z. [28] L. Cao, W. Lu, A. Mata, K. Nishinari, and Y. Fang. Egg-box model-based gelation of alginate and pectin: A review. Carbohydrate Polymers, 242:116389, Aug. 2020. DOI: 10.1016/j.carbpol.2020.116389.
29] E. Cetin, K. Aleksanyan Magden, Y. Zhou, and G. Yilmaz. Effect of hydrodynamic conditions on the formation and structure of aerobic granular sludge performing enhanced biological phosphorus removal. Water and Environment Journal, 36(1):56–66, Oct. 2021. DOI: 10.1111/wej.12754. [30] R. Chan and V. Chen. The effects of electrolyte concentration and ph on protein aggregation and deposition: critical flux and constant flux membrane filtration. Journal of Membrane Science, 185(2):177–192, Apr. 2001. DOI: 10.1016/s0376-7388(00)00645-1. [31] L. Chen, Q. Hu, X. Zhang, Z. Chen, Y. Wang, and S. Liu. Effects of salinity on the biological performance of anaerobic membrane bioreactor. Journal of Environmental Management, 238:263–273, May 2019. DOI: 10.1016/j. jenvman.2019.03.012. [32] W. Chen, P. Westerhoff, J. A. Leenheer, and K. Booksh. Fluorescence excitationemission matrix regional integration to quantify spectra for dissolved organic matter. Environmental Science and Technology, 37(24):5701–5710, Nov. 2003. DOI: 10.1021/es034354c. [33] W.-H. Chen, Y.-S. Chu, J.-L. Liu, and J.-S. Chang. Thermal degradation of carbohydrates, proteins and lipids in microalgae analyzed by evolutionary computation. Energy Conversion and Management, 160:209–219, Mar. 2018. DOI: 10.1016/j.enconman.2018.01.036. [34] Y.-P. Chen, P. Zhang, J.-S. Guo, F. Fang, X. Gao, and C. Li. Functional groups characteristics of EPS in biofilm growing on different carriers. Chemosphere, 92(6):633–638, July 2013. DOI: 10.1016/j.chemosphere.2013.01.059. [35] S. Chong, T. K. Sen, A. Kayaalp, and H. M. Ang. The performance enhancements of upflow anaerobic sludge blanket (UASB) reactors for domestic sludge treatment - A State-of-the-art review. Water Research, 46(11):3434–3470, July 2012. DOI: 10.1016/j.watres.2012.03.066. [36] L. A. Cole. Evolution of Chemical, Prokaryotic, and Eukaryotic Life, pages 93–99. Elsevier, Amsterdam, 2016. DOI: 10.1016/b978-0-12-809685-7. 00013-7. [37] S. Comte, G. Guibaud, and M. Baudu. Relations between extraction protocols for activated sludge extracellular polymeric substances (EPS) and complexation properties of Pb and Cd with EPS. Enzyme and Microbial Technology, 38(1- 2):246–252, Jan. 2006. DOI: 10.1016/j.enzmictec.2005.06.023. [38] Council of the European Union. Wastewater treatment, 2024. https://www. consilium.europa.eu/en/policies/wastewater-treatment/,[Accessed on 11/11/2024. [39] I. Cumpstey. Chemical modification of polysaccharides. ISRN Organic Chemistry, 2013:1–27, Sept. 2013. DOI: 10.1155/2013/417672.
[40] S. Dababat, G. Enaime, M. Wichern, and M. Lübken. Aerobic Granular Sludge: Perspectives for Excess Sludge Management and Resource Recovery. Chemie Ingenieur Technik, 95(12):1881–1896, Oct. 2023. DOI: 10.1002/cite. 202300093. [41] P. d’Abzac, F. Bordas, E. Joussein, E. van Hullebusch, P. Lens, and G. Guibaud. Characterization of the Mineral Fraction Associated to Extracellular Polymeric Substances (EPS) in Anaerobic Granular Sludges. Environmental Science and Technology, 44(1):412–418, Dec. 2009. DOI: 10.1021/es901912g. [42] Q. Dai, L. Ma, N. Ren, P. Ning, Z. Guo, L. Xie, and H. Gao. Investigation on extracellular polymeric substances, sludge flocs morphology, bound water release and dewatering performance of sewage sludge under pretreatment with modified phosphogypsum. Water Research, 142:337–346, Oct. 2018. DOI: 10.1016/j.watres.2018.06.009. [43] L. de Bruin, M. de Kreuk, H. van der Roest, C. Uijterlinde, and M. van Loosdrecht. Aerobic granular sludge technology: an alternative to activated sludge? Water Science and Technology, 49(11-12):1–7, June 2004. DOI: 10.2166/wst.2004.0790. [44] M. de Kreuk, J. Heijnen, and M. van Loosdrecht. Simultaneous COD, nitrogen, and phosphate removal by aerobic granular sludge. Biotechnology and Bioengineering, 90(6):761–769, Apr. 2005. DOI: 10.1002/bit.20470. [45] M. De Kreuk, B. McSwain, S. Bathe, S. Tay, N. Schwarzenbeck, and P. Wilderer. Discussion outcomes. Aerobic granular sludge, pages 155–169, 2005. [46] C. A. de Lemos Chernicharo. Anaerobic reactors. Number 4 in Biological Wastewater Treatment Series. IWA Publishing, London, 2007. [47] V. M. Dembitsky and M. Srebnik. Natural halogenated fatty acids: their analogues and derivatives. Progress in Lipid Research, 41(4):315–367, July 2002. DOI: 10.1016/s0163-7827(02)00003-6. [48] A. Demirbas, G. Edris, and W. M. Alalayah. Sludge production from municipal wastewater treatment in sewage treatment plant. Energy Sources, Part A: Recovery, Utilization, and Environmental Effects, 39(10):999–1006, apr 2017. DOI: 10.1080/15567036.2017.1283551. [49] K. I. Draget, G. Skjåk-Bræk, and O. Smidsrød. Alginate based new materials. International Journal of Biological Macromolecules, 21(1-2):47–55, Aug. 1997. DOI: 10.1016/s0141-8130(97)00040-8. [50] C.-D. Dubé and S. R. Guiot. Characterization of the protein fraction of the extracellular polymeric substances of three anaerobic granular sludges. AMB Express, 9(1), Feb. 2019. DOI: 10.1186/s13568-019-0746-0.
[51] M. Dubois, K. Gilles, J. K. Hamilton, P. A. Rebers, and F. Smith. A colorimetric method for the determination of sugars. Nature, 168(4265):167–167, July 1951. DOI: 10.1038/168167a0. [52] A. K. Dwivedi. Researches in water pollution: A review. International Research Journal of Natural and Applied Sciences, 4(1):118–142, 2017. [53] M. Eggersdorfer and A. Wyss. Carotenoids in human nutrition and health. Archives of Biochemistry and Biophysics, 652:18–26, Aug. 2018. DOI: 10.1016/ j.abb.2018.06.001. [54] H. El Farissi, R. Lakhmiri, A. Albourine, M. Safi, and O. Cherkaoui. Removal of RR-23 dye from industrial textile wastewater by adsorption on cistus ladaniferus seeds and their biochar. 7, 11 2017. [55] J. Elleuch, M. Drira, I. Ghribi, F. Hadjkacem, G. Pierre, C. Causserand, H. Khemakhem, P. Michaud, I. Fendri, and S. Abdelkafi. Amphora coffeiformis extracellular polymeric substances and their potential applications in lead removal. Antonie van Leeuwenhoek, 118(3), Feb. 2025. DOI: 10.1007/s10482-024-02057-6. [56] K. Ellis. Slow sand filtration as a technique for the tertiary treatment of municipal sewages. Water Research, 21(4):403–410, Apr. 1987. DOI: 10.1016/ 0043-1354(87)90187-4. [57] H. H. P. Fang and X. S. Jia. Extraction of extracellular polymer from anaerobic sludges. Biotechnology Techniques, 10(11):803–808, Nov. 1996. DOI: 10.1007/ bf00154662. [58] G. Feijoo, M. Soto, R. Mendez, and J. M. Lema. Sodium inhibition in the anaerobic digestion process: Antagonism and adaptation phenomena. Enzyme and Microbial Technology, 17(2):180–188, Feb. 1995. DOI: 10.1016/0141-0229(94)00011-f. [59] S. Felz. Structural Extracellular Polymeric Substances from Aerobic Granular Sludge. PhD thesis, Delft University of Technology, 2019. DOI: 10.4233/UUID: 93E702D1-92B2-4025-AB57-6D2C141ED14D. [60] S. Felz, H. Kleikamp, J. Zlopasa, M. C. van Loosdrecht, and Y. Lin. Impact of metal ions on structural EPS hydrogels from aerobic granular sludge. Biofilm, 2:100011, Dec. 2020. DOI: 10.1016/j.bioflm.2019.100011. [61] C. Feng, T. Lotti, R. Canziani, Y. Lin, C. Tagliabue, and F. Malpei. Extracellular biopolymers recovered as raw biomaterials from waste granular sludge and potential applications: A critical review. Science of The Total Environment, 753:142051, Jan. 2021. DOI: 10.1016/j.scitotenv.2020.142051.
[62] R. Ferrentino, M. Langone, L. Fiori, and G. Andreottola. Full-Scale Sewage Sludge Reduction Technologies: A Review with a Focus on Energy Consumption. Water, 15(4):615, Feb. 2023. DOI: 10.3390/w15040615. [63] H.-C. Flemming and J. Wingender. The biofilm matrix. Nature Reviews Microbiology, 8(9):623–633, Aug. 2010. DOI: 10.1038/nrmicro2415. [64] D. Focht and A. Chang. Nitrification and Denitrification Processes Related to Waste Water Treatment, pages 153–186. Elsevier, Amsterdam, 1975. DOI: 10.1016/s0065-2164(08)70428-3. [65] A. Foglia, C. Akyol, N. Frison, E. Katsou, A. L. Eusebi, and F. Fatone. Longterm operation of a pilot-scale anaerobic membrane bioreactor (anmbr) treating high salinity low loaded municipal wastewater in real environment. Separation and Purification Technology, 236:116279, Apr. 2020. DOI: 10.1016/j.seppur. 2019.116279. [66] K. Frank. Microbiology in Clinical Pathology, pages 3237–3268. Elsevier, Amsterdam, 2014. DOI: 10.1016/b978-0-12-386456-7.06304-8. [67] B. Frølund, R. Palmgren, K. Keiding, and P. H. Nielsen. Extraction of extracellular polymers from activated sludge using a cation exchange resin. Water Research, 30(8):1749–1758, Aug. 1996. DOI: 10.1016/0043-1354(95) 00323-1. [68] M. Gagliano, S. Ismail, A. Stams, C. Plugge, H. Temmink, and J. Van Lier. Biofilm formation and granule properties in anaerobic digestion at high salinity. Water Research, 121:61–71, Sept. 2017. DOI: 10.1016/j.watres.2017.05. 016. [69] M. C. Gagliano, T. R. Neu, U. Kuhlicke, D. Sudmalis, H. Temmink, and C. M. Plugge. EPS Glycoconjugate Profiles Shift as Adaptive Response in Anaerobic Microbial Granulation at High Salinity. Frontiers in Microbiology, 9:1423, July 2018. DOI: 10.3389/fmicb.2018.01423. [70] D. Gao, L. Liu, H. Liang, and W.-M. Wu. Aerobic granular sludge: characterization, mechanism of granulation and application to wastewater treatment. Critical Reviews in Biotechnology, 31(2):137–152, Oct. 2010. DOI: 10.3109/07388551.2010.497961. [71] R. Geetha Bai and R. Tuvikene. Potential Antiviral Properties of Industrially Important Marine Algal Polysaccharides and Their Significance in Fighting a Future Viral Pandemic. Viruses, 13(9):1817, Sept. 2021. DOI: 10.3390/ v13091817. [72] R. Gehr. Removal of extracellular material techniques and pitfalls. Water Research, 17(12):1743–1748, 1983. DOI: 10.1016/0043-1354(83)90195-1.
[73] K. V. Gernaey, M. C. van Loosdrecht, M. Henze, M. Lind, and S. B. Jorgensen. Activated sludge wastewater treatment plant modelling and simulation: state of the art. Environmental Modelling and Software, 19(9):763–783, Sept. 2004. DOI: 10.1016/j.envsoft.2003.03.005. [74] A. G. Geyik and F. Cecen. Variations in extracellular polymeric substances (EPS) during adaptation of activated sludges to new feeding conditions. International Biodeterioration and Biodegradation, 105:137–145, Nov. 2015. DOI: 10.1016/j.ibiod.2015.08.021. [75] R. Gheorghita Puscaselu, A. Lobiuc, M. Dimian, and M. Covasa. Alginate: From food industry to biomedical applications and management of metabolic disorders. Polymers, 12(10):2417, Oct. 2020. DOI: 10.3390/polym12102417. [76] E. Gomez-Ordonez and P. Ruperez. FTIR-ATR spectroscopy as a tool for polysaccharide identification in edible brown and red seaweeds. Food Hydrocolloids, 25(6):1514–1520, Aug. 2011. DOI: 10.1016/j.foodhyd.2011.02.009. [77] G. Gonzalez-Gil, L. Thomas, A.-H. Emwas, P. N. L. Lens, and P. E. Saikaly. NMR and MALDI-TOF MS based characterization of exopolysaccharides in anaerobic microbial aggregates from full-scale reactors. Scientific Reports, 5(1), Sept. 2015. DOI: 10.1038/srep14316. [78] G. Goswami, S. Chaudhuri, and D. Dutta. Studies on the stability of a carotenoid produced by a novel isolate using low cost agro-industrial residue and its application in different model systems. LWT - Food Science and Technology, 63(1):780–790, Sept. 2015. DOI: 10.1016/j.lwt.2015.03.017. [79] G. T. Grant, E. R. Morris, D. A. Rees, P. J. Smith, and D. Thom. Biological interactions between polysaccharides and divalent cations: The egg-box model. FEBS Letters, 32(1):195–198, May 1973. DOI: 10.1016/0014-5793(73) 80770-7. [80] G. Guibaud, S. Comte, F. Bordas, S. Dupuy, and M. Baudu. Comparison of the complexation potential of extracellular polymeric substances (EPS), extracted from activated sludges and produced by pure bacteria strains, for cadmium, lead and nickel. Chemosphere, 59(5):629–638, Apr. 2005. DOI: 10.1016/j.chemosphere.2004.10.028. [81] S. R. Guiot, A. Pauss, and J. W. Costerton. A structured model of the anaerobic granule consortium. Water Science and Technology, 25(7):1–10, Apr. 1992. DOI: 10.2166/wst.1992.0133. [82] J. Guo, J. Zhou, D. Wang, C. Tian, P. Wang, M. Salah Uddin, and H. Yu. Biocalalyst effects of immobilized anthraquinone on the anaerobic reduction of azo dyes by the salt-tolerant bacteria. Water Research, 41(2):426–432, Jan. 2007. DOI: 10.1016/j.watres.2006.10.022.
[83] X. Guo, J. Liu, and B. Xiao. Evaluation of the damage of cell wall and cell membrane for various extracellular polymeric substance extractions of activated sludge. Journal of Biotechnology, 188:130–135, Oct. 2014. DOI: 10.1016/j.jbiotec.2014.08.025. [84] X. Guo, Y. Miao, B. Wu, L. Ye, H. Yu, S. Liu, and X.-x. Zhang. Correlation between microbial community structure and biofouling as determined by analysis of microbial community dynamics. Bioresource Technology, 197:99–105, Dec. 2015. DOI: 10.1016/j.biortech.2015.08.049. [85] J. Gupta, R. Rathour, C. L. Dupont, D. Kaul, and I. S. Thakur. Genomic insights into waste valorized extracellular polymeric substances (EPS) produced by Bacillus sp. ISTL8. Environmental Research, 192:110277, Jan. 2021. DOI: 10.1016/j.envres.2020.110277. [86] M. Hamoda, I. Al-Ghusain, and N. Al-Mutairi. Sand filtration of wastewater for tertiary treatment and water reuse. Desalination, 164(3):203–211, Apr. 2004. DOI: 10.1016/s0011-9164(04)00189-4. [87] S. He, L. Feng, W. Zhao, J. Li, Q. Zhao, and L. Wei. Composition and molecular structure analysis of hydrophilic/hydrophobic extracellular polymeric substances (EPS) with impacts on sludge dewaterability. Chemical Engineering Journal, 462:142234, Apr. 2023. DOI: 10.1016/j.cej.2023.142234. [88] A.-M. Hermansson, E. Eriksson, and E. Jordansson. Effects of potassium, sodium and calcium on the microstructure and rheological behaviour of kappacarrageenan gels. Carbohydrate Polymers, 16(3):297–320, Jan. 1991. DOI: 10.1016/0144-8617(91)90115-s. [89] M. J. Higgins and J. T. Novak. Characterization of exocellular protein and its role in bioflocculation. Journal of Environmental Engineering, 123(5):479–485, May 1997. DOI: 10.1061/(asce)0733-9372(1997)123:5(479). [90] P.-N. Hong, R. Honda, M. Noguchi, and T. Ito. Optimum selection of extraction methods of extracellular polymeric substances in activated sludge for effective extraction of the target components. Biochemical Engineering Journal, 127:136– 146, Nov. 2017. DOI: 10.1016/j.bej.2017.08.002. [91] T. Hong, J.-Y. Yin, S.-P. Nie, and M.-Y. Xie. Applications of infrared spectroscopy in polysaccharide structural analysis: Progress, challenge and perspective. Food Chemistry: X, 12:100168, Dec. 2021. DOI: 10.1016/j.fochx.2021. 100168. [92] N. Horan and C. Eccles. Purification and characterization of extracellular polysaccharide from activated sludges. Water Research, 20(11):1427–1432, Nov. 1986. DOI: 10.1016/0043-1354(86)90142-9.
[93] B. Hu, L. Du, and S. Matsukawa. NMR study on the network structure of a mixed gel of kappa and iota carrageenans. Carbohydrate Polymers, 150:57–64, Oct. 2016. DOI: 10.1016/j.carbpol.2016.04.112. [94] L. Hulshoff Pol, S. de Castro Lopes, G. Lettinga, and P. Lens. Anaerobic sludge granulation. Water Research, 38(6):1376–1389, Mar. 2004. DOI: 10. 1016/j.watres.2003.12.002. [95] S. Ismail, C. de La Parra, H. Temmink, and J. van Lier. Extracellular polymeric substances (EPS) in upflow anaerobic sludge blanket (UASB) reactors operated under high salinity conditions. Water Research, 44(6):1909–1917, Mar. 2010. DOI: 10.1016/j.watres.2009.11.039. [96] H. Iwamura. Organic reactions in sub- and supercritical water in the absence of any added catalyst. The Journal of Research Institute of Science and Technology, Nihon University, 132:1–9, 01 2014. [97] P. U. Iyare, S. K. Ouki, and T. Bond. Microplastics removal in wastewater treatment plants: a critical review. Environmental Science: Water Research and Technology, 6(10):2664–2675, 2020. DOI: 10.1039/d0ew00397b. [98] P. Jaibiba, S. Naga Vignesh, and S. Hariharan. Working principle of typical bioreactors, pages 145–173. Elsevier, Amsterdam, 2020. DOI: 10.1016/b978-0-12-821264-6.00010-3. [99] H.-L. Jiang, J.-H. Tay, and S. T.-L. Tay. Changes in structure, activity and metabolism of aerobic granules as a microbial response to high phenol loading. Applied Microbiology and Biotechnology, (5):602–608, May 2004. [100] P. B. Jordal, M. S. Dueholm, P. Larsen, S. V. Petersen, J. J. Enghild, G. Christiansen, P. Hø jrup, P. H. Nielsen, and D. E. Otzen. Widespread Abundance of Functional Bacterial Amyloid in Mycolata and Other Gram-Positive Bacteria. Applied and Environmental Microbiology, 75(12):4101–4110, June 2009. DOI: 10.1128/aem.02107-08. [101] D. Kalita, P. Konwar, D. Sahu, S. Baruah, and S. Barua. Fluoropolymer nanocomposites: introduction, fundamental properties, and high-performance applications, pages 79–120. Elsevier, Amsterdam, 2023. DOI: 10.1016/ b978-0-323-95335-1.00023-2. [102] I. Karakas, S. B. Sam, E. Cetin, E. Dulekgurgen, and G. Yilmaz. Resource recovery from an aerobic granular sludge process treating domestic wastewater. Journal of Water Process Engineering, 34:101148, Apr. 2020. DOI: 10.1016/j. jwpe.2020.101148. [103] K. Kaviyarasan. Application of UASB Reactor in Industrial Wastewater Treatment - A Review. volume 5, 2014. ISSN: 2229-5518.
[104] J. Keller and K. Hartley. Greenhouse gas production in wastewater treatment: process selection is the major factor. Water Science and Technology, 47(12):43– 48, June 2003. DOI: 10.2166/wst.2003.0626. [105] N. K. Kim, N. Mao, R. Lin, D. Bhattacharyya, M. C. van Loosdrecht, and Y. Lin. Flame retardant property of flax fabrics coated by extracellular polymeric substances recovered from both activated sludge and aerobic granular sludge. Water Research, 170:115344, Mar. 2020. DOI: 10.1016/j.watres. 2019.115344. [106] P. J. A. Kleinman, A. N. Sharpley, R. W. McDowell, D. N. Flaten, A. R. Buda, L. Tao, L. Bergstrom, and Q. Zhu. Managing agricultural phosphorus for water quality protection: principles for progress. Plant and Soil, 349(1-2):169–182, July 2011. DOI: 10.1007/s11104-011-0832-9. [107] T. Kobayashi, K. Abe, K. Asai, K. Gomi, P. R. Juvaddi, M. Kato, K. Kitamoto, M. Takeuchi, and M. Machida. Genomics of aspergillus oryzae. Bioscience, Biotechnology, and Biochemistry, 71(3):646–670, Mar. 2007. DOI: 10.1271/ bbb.60550. [108] K. Krysiak-Baltyn, R. Cavalida, B. Thwaites, P. J. Reeve, P. J. Scales, B. Van den Akker, L. Ong, G. J. Martin, A. D. Stickland, and S. L. Gras. Comparison of physical characteristics and dewatering behaviour between granular and floccular sludges generated from the same sewage source. Journal of Water Process Engineering, 29:100785, June 2019. DOI: 10.1016/j.jwpe.2019.100785. [109] P. Larsen, J. L. Nielsen, D. Otzen, and P. H. Nielsen. Amyloid-Like Adhesins Produced by Floc-Forming and Filamentous Bacteria in Activated Sludge. Applied and Environmental Microbiology, 74(5):1517–1526, Mar. 2008. DOI: 10.1128/aem.02274-07. [110] W. C. Lay, Y. Liu, and A. G. Fane. Impacts of salinity on the performance of high retention membrane bioreactors for water reclamation: A review. Water Research, 44(1):21–40, Jan. 2010. DOI: 10.1016/j.watres.2009.09.026. [111] T. M. Le, Y. Lin, W.-Q. Zhuang, K. Jayaraman, and N. K. Kim. Effects of Extraction Methods on the Thermal Stability of Extracellular Polymeric Substances-Based Biomaterials from Wastewater Sludge. Environmental Science & Technology, 59(8):4165–4177, Feb. 2025. DOI: 10.1021/acs.est. 4c10329. [112] S. Le Borgne, D. Paniagua, and R. Vazquez-Duhalt. Biodegradation of organic pollutants by halophilic bacteria and archaea. Microbial Physiology, 15(2-3):74– 92, 2008. DOI: 10.1159/000121323. [113] K. Y. Lee and D. J. Mooney. Alginate: Properties and biomedical applications. Progress in Polymer Science, 37(1):106–126, Jan. 2012. DOI: 10.1016/j. progpolymsci.2011.06.003.
114] N. M. Lee and T. Welander. Reducing sludge production in aerobic wastewater treatment through manipulation of the ecosystem. Water Research, 30(8):1781– 1790, Aug. 1996. DOI: 10.1016/0043-1354(96)00059-0. [115] O. Lefebvre and R. Moletta. Treatment of organic pollution in industrial saline wastewater: A literature review. Water Research, 40(20):3671–3682, Dec. 2006. DOI: 10.1016/j.watres.2006.08.027. [116] O. Lefebvre, S. Quentin, M. Torrijos, J. J. Godon, J. P. Delgenes, and R. Moletta. Impact of increasing nacl concentrations on the performance and community composition of two anaerobic reactors. Applied Microbiology and Biotechnology, 75(1):61–69, May 2007. DOI: 10.1007/s00253-006-0799-2. [117] J. Lei, X. Zhang, Y. Liu, X. Wang, G. Li, T. Feng, and H. Zhong. Adsorption Characteristics between Different Sizes of Microplastics and EPS Fractions of Anaerobic Granular Sludge. Polish Journal of Environmental Studies, 33(3):2141–2148, Jan. 2024. DOI: 10.15244/pjoes/173167. [118] G. Lettinga, A. F. M. van Velsen, S. W. Hobma, W. de Zeeuw, and A. Klapwijk. Use of the upflow sludge blanket (usb) reactor concept for biological wastewater treatment, especially for anaerobic treatment. Biotechnology and Bioengineering, 22(4):699–734, Apr. 1980. DOI: 10.1002/bit.260220402. [119] J. Li, K. F. See, and J. Chi. Water resources and water pollution emissions in China’s industrial sector: A green-biased technological progress analysis. Journal of Cleaner Production, 229:1412–1426, Aug. 2019. DOI: 10.1016/j. jclepro.2019.03.216. [120] N. Li, D. Wei, S. Wang, L. Hu, W. Xu, B. Du, and Q. Wei. Comparative study of the role of extracellular polymeric substances in biosorption of ni(ii) onto aerobic/anaerobic granular sludge. Journal of Colloid and Interface Science, 490:754–761, Mar. 2017. DOI: 10.1016/j.jcis.2016.12.006. [121] W.-W. Li and H.-Q. Yu. Insight into the roles of microbial extracellular polymer substances in metal biosorption. Bioresource Technology, 160:15–23, May 2014. DOI: 10.1016/j.biortech.2013.11.074. [122] S. J. Lim and T.-H. Kim. Applicability and trends of anaerobic granular sludge treatment processes. Biomass and Bioenergy, 60:189–202, Jan. 2014. DOI: 10.1016/j.biombioe.2013.11.011. [123] Y. Lin, K. Nierop, E. Girbal-Neuhauser, M. Adriaanse, and M. van Loosdrecht. Sustainable polysaccharide-based biomaterial recovered from waste aerobic granular sludge as a surface coating material. Sustainable Materials and Technologies, 4:24–29, July 2015. DOI: 10.1016/j.susmat.2015.06.002
[124] Y. M. Lin, L. Wang, Z. M. Chi, and X. Y. Liu. Bacterial alginate role in aerobic granular bioparticles formation and settleability improvement. Separation Science and Technology, 43(7):1642–1652, May 2008. DOI: 10.1080/01496390801973805. [125] S. Liparoti, V. Speranza, and F. Marra. Alginate hydrogel: The influence of the hardening on the rheological behaviour. Journal of the Mechanical Behavior of Biomedical Materials, 116:104341, Apr. 2021. DOI: 10.1016/j.jmbbm.2021. 104341. [126] H. Liu and H. H. Fang. Extraction of extracellular polymeric substances (EPS) of sludges. Journal of Biotechnology, 95(3):249–256, May 2002. DOI: 10.1016/s0168-1656(02)00025-1. [127] L. Liu, D.-W. Gao, M. Zhang, and Y. Fu. Comparison of ca2+ and mg2+ enhancing aerobic granulation in sbr. Journal of Hazardous Materials, 181(1- 3):382–387, Sept. 2010. DOI: 10.1016/j.jhazmat.2010.05.021. [128] M. Liu, J. J. Gill, R. Young, and E. J. Summer. Bacteriophages of wastewater foaming-associated filamentous gordonia reduce host levels in raw activated sludge. Scientific Reports, 5(1), Sept. 2015. DOI: 10.1038/srep13754. [129] X. Liu, J. Liu, D. Deng, R. Li, C. Guo, J. Ma, and M. Chen. Investigation of extracellular polymeric substances (EPS) in four types of sludge: Factors influencing EPS properties and sludge granulation. Journal of Water Process Engineering, 40:101924, Apr. 2021. DOI: 10.1016/j.jwpe.2021.101924. [130] X. Liu, Q. Pei, H. Han, H. Yin, M. Chen, C. Guo, J. Li, and H. Qiu. Functional analysis of extracellular polymeric substances (EPS) during the granulation of aerobic sludge: Relationship among EPS, granulation and nutrients removal. Environmental Research, 208:112692, May 2022. DOI: 10.1016/j.envres. 2022.112692. [131] Y. Liu and H. H. P. Fang. Influences of Extracellular Polymeric Substances (EPS) on Flocculation, Settling, and Dewatering of Activated Sludge. Critical Reviews in Environmental Science and Technology, 33(3):237–273, July 2003. DOI: 10.1080/10643380390814479. [132] Y. Liu and J.-H. Tay. State of the art of biogranulation technology for wastewater treatment. Biotechnology Advances, 22(7):533–563, Sept. 2004. DOI: 10.1016/j.biotechadv.2004.05.001. [133] S. Lorenzen. The isolated effect of particle surface charge on filter cake properties. The PhD Series of the Faculty of Engineering and Science, 2016. DOI: 10.5278/VBN.PHD.ENGSCI.00185. [134] T. Lotti, E. Carretti, D. Berti, C. Montis, S. Del Buffa, C. Lubello, C. Feng, and F. Malpei. Hydrogels formed by anammox extracellular polymeric substances: structural and mechanical insights. Scientific Reports, 9(1), Aug. 2019. DOI: 10.1038/s41598-019-47987-8. [135] O. H. Lowry, N. Rosebrough, A. L. Farr, and R. J. Randall. Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193(1):265–275, Nov. 1951. DOI: 10.1016/s0021-9258(19)52451-6. [136] H. Lu, K. Chandran, and D. Stensel. Microbial ecology of denitrification in biological wastewater treatment. Water Research, 64:237–254, Nov. 2014. DOI: 10.1016/j.watres.2014.06.042. [137] L. Lundqvist. Structural and interaction studies of polysaccharides by NMR spectroscopy. PhD thesis, Swedish University of Agricultural Sciences, 2015. [138] F. A. MacLeod, S. R. Guiot, and J. W. Costerton. Layered structure of bacterial aggregates produced in an upflow anaerobic sludge bed and filter reactor. Applied and Environmental Microbiology, 56(6):1598–1607, June 1990. DOI: 10.1128/aem.56.6.1598-1607.1990. [139] N. R. Maddela, Z. Zhou, Z. Yu, S. Zhao, and F. Meng. Functional Determinants of Extracellular Polymeric Substances in Membrane Biofouling: Experimental Evidence from Pure-Cultured Sludge Bacteria. Applied and Environmental Microbiology, 84(15), Aug. 2018. DOI: 10.1128/aem.00756-18. [140] A. Mahardika, A. B. Susanto, R. Pramesti, H. Matsuyoshi, B. B. Andriana, Y. Matsuda, and H. Sato. Application of imaging Raman spectroscopy to study the distribution of Kappa carrageenan in the seaweed Kappaphycus alvarezii. Journal of Applied Phycology, 31(2):1383–1390, Sept. 2018. DOI: 10.1007/s10811-018-1618-8. [141] J. Mallevialle, P. E. Odendaal, and M. R. Wiesner. Water treatment membrane processes. McGraw-Hill Professional, New York City, 1996. [142] A. T. Mansour, A. E. Alprol, K. M. Abualnaja, H. S. El-Beltagi, K. M. A. Ramadan, and M. Ashour. The Using of Nanoparticles of Microalgae in Remediation of Toxic Dye from Industrial Wastewater: Kinetic and Isotherm Studies. Materials, 15(11):3922, May 2022. DOI: 10.3390/ma15113922. [143] M. Mecozzi, M. Pietroletti, M. Scarpiniti, R. Acquistucci, and M. E. Conti. Monitoring of marine mucilage formation in Italian seas investigated by infrared spectroscopy and independent component analysis. Environmental Monitoring and Assessment, 184(10):6025–6036, Oct. 2011. DOI: 10.1007/ s10661-011-2400-4. [144] J. N. Meegoda, B. Li, K. Patel, and L. B. Wang. A review of the processes, parameters, and optimization of anaerobic digestion. International journal of environmental research and public health, 15, Oct. 2018. DOI 10.3390/ ijerph15102224.
[145] L. Mertens. Can bioaugmentation with aerobic fungal pellets enhance the start-up of an anaerobic digester treating saline wastewater with a non-salineacclimated inoculum? Master’s thesis, KULeuven, 2025. [146] U. Metzger, U. Lankes, K. Fischpera, and F. H. Frimmel. The concentration of polysaccharides and proteins in EPS of Pseudomonas putida and Aureobasidum pullulans as revealed by 13C CPMAS NMR spectroscopy. Applied Microbiology and Biotechnology, 85(1):197–206, Oct. 2009. DOI: 10.1007/s00253-009-2218-y. [147] T. Mino, M. van Loosdrecht, and J. Heijnen. Microbiology and biochemistry of the enhanced biological phosphate removal process. Water Research, 32(11):3193–3207, Nov. 1998. DOI: 10.1016/s0043-1354(98)00129-8. [148] E. R. Morris, D. A. Rees, and G. Robinson. Cation-specific aggregation of carrageenan helices: Domain model of polymer gel structure. Journal of Molecular Biology, 138(2):349–362, Apr. 1980. DOI: 10.1016/0022-2836(80) 90291-0. [149] J. D. Munoz Sierra, M. J. Oosterkamp, W. Wang, H. Spanjers, and J. B. van Lier. Comparative performance of upflow anaerobic sludge blanket reactor and anaerobic membrane bioreactor treating phenolic wastewater: Overcoming high salinity. Chemical Engineering Journal, 366:480–490, June 2019. DOI: 10.1016/j.cej.2019.02.097. [150] S. Naidoo and A. Olaniran. Treated Wastewater Effluent as a Source of Microbial Pollution of Surface Water Resources. International Journal of Environmental Research and Public Health, 11(1):249–270, Dec. 2013. DOI: 10.3390/ijerph110100249. [151] NASA. Why europa: Ingredients for life, 2022. https://europa.nasa.gov/ why-europa/ingredients-for-life/#:~:text=Scientists%20say%20we% 20should%20look,life%20takes%20time%20to%20develop. [Accessed: 11 November 2024]. [152] L. N. Nguyen, A. Q. Nguyen, and L. D. Nghiem. Microbial Community in Anaerobic Digestion System: Progression in Microbial Ecology, pages 331–355. Springer Singapore, Nov. 2018. DOI: 10.1007/978-981-13-3259-31-5. [153] S.-Q. Ni, N. Sun, H. Yang, J. Zhang, and H. H. Ngo. Distribution of extracellular polymeric substances in anammox granules and their important roles during anammox granulation. Biochemical Engineering Journal, 101:126–133, Sept. 2015. DOI: 10.1016/j.bej.2015.05.014. [154] Y. Nie, Z. Wang, W. Wang, Z. Zhou, Y. Kong, and J. Ma. Bio-flocculation of microcystis aeruginosa by using fungal pellets of aspergillus oryzae: Performance and mechanism. Journal of Hazardous Materials, 439:129606, Oct. 2022. DOI: 10.1016/j.jhazmat.2022.129606.
[155] P. H. Nielsen, A. M. Saunders, A. A. Hansen, P. Larsen, and J. L. Nielsen. Microbial communities involved in enhanced biological phosphorus removal from wastewater-a model system in environmental biotechnology. Current Opinion in Biotechnology, 23(3):452–459, June 2012. DOI: 10.1016/j.copbio. 2011.11.027. [156] O. Nowak. Optimizing the use of sludge treatment facilities at municipal wwtps. Journal of Environmental Science and Health, Part A, 41(9):1807–1817, Sept. 2006. DOI: 10.1080/10934520600778986. [157] B. Pagliaccia. Insights on the recovery, characterization and valorization of Extracellular Polymeric substances (EPS) from granular sludge applied in innovative wastewater treatment systems. PhD thesis, Universite de Toulouse, 2022. [158] B. Pagliaccia, R. Campo, E. Carretti, M. Severi, C. Lubello, and T. Lotti. Towards resource recovery-oriented solutions in agriculture exploiting structural extracellular polymeric substances (sEPS) extracted from aerobic granular sludge (AGS). Chemical Engineering Journal, 485:149819, Apr. 2024. DOI: 10.1016/j.cej.2024.149819. [159] B. Pagliaccia, E. Carretti, M. Severi, D. Berti, C. Lubello, and T. Lotti. Heavy metal biosorption by Extracellular Polymeric Substances (EPS) recovered from anammox granular sludge. Journal of Hazardous Materials, 424:126661, Feb. 2022. DOI: 10.1016/j.jhazmat.2021.126661. [160] L. Passauer, T. Hallas, E. Baucker, G. Ciesielski, S. Lebioda, and U. Hamer. Biodegradation of hydrogels from oxyethylated lignins in model soils. ACS Sustainable Chemistry and Engineering, 3(9):1955–1964, Aug. 2015. DOI: 10.1021/acssuschemeng.5b00139. [161] M. Patziger, H. Kainz, M. Hunze, and J. Jozsa. Influence of secondary settling tank performance on suspended solids mass balance in activated sludge systems. Water Research, 46(7):2415–2424, May 2012. DOI: 10.1016/j.watres.2012. 02.007. [162] S. Peng, A. Hu, J. Ai, W. Zhang, and D. Wang. Changes in molecular structure of extracellular polymeric substances (EPS) with temperature in relation to sludge macro-physical properties. Water Research, 201:117316, Aug. 2021. DOI: 10.1016/j.watres.2021.117316. [163] T. Peng, Y. Wang, J. Wang, F. Fang, P. Yan, and Z. Liu. Effect of different forms and components of EPS on sludge aggregation during granulation process of aerobic granular sludge. Chemosphere, 303:135116, Sept. 2022. DOI: 10. 1016/j.chemosphere.2022.135116.
[164] O. Pidoux, J.-N. Argenson, V. Jacomo, and M. Drancourt. Molecular identification of a dietzia maris hip prosthesis infection isolate. Journal of Clinical Microbiology, 39(7):2634–2636, July 2001. DOI: 10.1128/jcm.39.7.2634-2636. 2001. [165] D. Pooja, P. Kumar, P. Singh, and S. Patil. Sensors in Water Pollutants Monitoring: Role of Material. Springer Singapore, 2020. DOI: 10.1007/ 978-981-15-0671-0. [166] C. Proikakis, N. Mamouzelos, P. Tarantili, and A. Andreopoulos. Swelling and hydrolytic degradation of poly(d,l-lactic acid) in aqueous solutions. Polymer Degradation and Stability, 91(3):614–619, Mar. 2006. DOI: 10.1016/j. polymdegradstab.2005.01.060. [167] M. Pronk, M. de Kreuk, B. de Bruin, P. Kamminga, R. Kleerebezem, and M. van Loosdrecht. Full scale performance of the aerobic granular sludge process for sewage treatment. Water Research, 84:207–217, Nov. 2015. DOI: 10.1016/j.watres.2015.07.011. [168] J. Prosser. Autotrophic Nitrification in Bacteria. Elsevier, Amsterdam, 1990. DOI: 10.1016/s-2911(08)60112-5. [169] S. Ramakrishnan and R. K. Prud’homme. Effect of solvent quality and ions on the rheology and gelation of κ-carrageenan. Journal of Rheology, 44(4):885–896, July 2000. DOI: 10.1122/1.551119. [170] K. Rasmussen, C. Reilly, Y. Li, and R. S. Jones. Realtime imaging of antibiofilm effects using cp oct. Biotechnology and Bioengineering, 113(1):198–205, Sept. 2015. DOI: 10.1002/bit.25701. [171] C. Reddy, R. Ghai, Rashmi, and V. Kalia. Polyhydroxyalkanoates: an overview. Bioresource Technology, 87(2):137–146, Apr. 2003. DOI: 10.1016/ s0960-8524(02)00212-2. [172] E. Reid, X. Liu, and S. Judd. Effect of high salinity on activated sludge characteristics and membrane permeability in an immersed membrane bioreactor. Journal of Membrane Science, 283(1-2):164–171, Oct. 2006. DOI: 10.1016/j.memsci.2006.06.021. [173] X. Ren, D. Yu, S. Han, and Y. Feng. Thermolysis of recombinant escherichia coli for recovering a thermostable enzyme. Biochemical Engineering Journal, 33(1):94–98, Jan. 2007. DOI: 10.1016/j.bej.2006.09.017. [174] M. Rigoulet, C. Bouchez, P. Paumard, S. Ransac, S. Cuvellier, S. DuvezinCaubet, J. Mazat, and A. Devin. Cell energy metabolism: An update. Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1861(11):148276, Nov. 2020. DOI: 10.1016/j.bbabio.2020.148276.
[175] R. Risi, L. Manti, G. Perna, M. Lasalvia, V. Capozzi, I. Delfino, and M. Lepore. X-ray radiation-induced effects in human mammary epithelial cells investigated by raman microspectroscopy. In J. Popp, W. Drexler, V. V. Tuchin, and D. L. Matthews, editors, Biophotonics: Photonic Solutions for Better Health Care III, volume 8427, page 84272E. SPIE, June 2012. DOI: 10.1117/12.921389. [176] L. Rizzo, A. Fiorentino, and A. Anselmo. Advanced treatment of urban wastewater by uv radiation: Effect on antibiotics and antibiotic-resistant e. coli strains. Chemosphere, 92(2):171–176, June 2013. DOI: 10.1016/j. chemosphere.2013.03.021. [177] R. Rupert, K. F. Rodrigues, V. Y. Thien, and W. T. L. Yong. Carrageenan From Kappaphycus alvarezii (Rhodophyta, Solieriaceae): Metabolism, Structure, Production, and Application. Frontiers in Plant Science, 13, May 2022. DOI: 10.3389/fpls.2022.859635. [178] S. Saji, A. Hebden, P. Goswami, and C. Du. A Brief Review on the Development of Alginate Extraction Process and Its Sustainability. Sustainability, 14(9):5181, Apr. 2022. DOI: 10.3390/su14095181. [179] M. Sajjad and K. S. Kim. Extraction of extracellular polymeric substances from activated sludge using sodium oxalate. International Journal of Environmental Science and Technology, 13(7):1697–1706, May 2016. DOI: 10.1007/s13762-016-1004-5. [180] H. Sanawar, I. Pinel, N. Farhat, S. Bucs, J. Zlopasa, J. Kruithof, G. Witkamp, M. van Loosdrecht, and J. Vrouwenvelder. Enhanced biofilm solubilization by urea in reverse osmosis membrane systems. Water Research X, 1:100004, Dec. 2018. DOI: 10.1016/j.wroa.2018.10.001. [181] C. M. Santegoeds, L. R. Damgaard, G. Hesselink, J. Zopfi, P. Lens, G. Muyzer, and D. de Beer. Distribution of sulfate-reducing and methanogenic bacteria in anaerobic aggregates determined by microsensor and molecular analyses. Applied and Environmental Microbiology, 65(10):4618–4629, Oct. 1999. DOI: 10.1128/aem.65.10.4618-4629.1999. [182] C. M. Schambeck, E. Girbal-Neuhauser, L. Böni, P. Fischer, Y. Bessière, E. Paul, R. H. R. da Costa, and N. Derlon. chemical and physical properties of alginate-like exopolymers of aerobic granules and flocs produced from different wastewaters. Bioresource Technology, 312:123632, Sept. 2020. DOI: 10.1016/ j.biortech.2020.123632. [183] T. Schmid, A. Messmer, B.-S. Yeo, W. Zhang, and R. Zenobi. Towards chemical analysis of nanostructures in biofilms II: tip-enhanced Raman spectroscopy of alginates. Analytical and Bioanalytical Chemistry, 391(5):1907–1916, Apr. 2008. DOI: 10.1007/s00216-008-2101-1.
[184] I. Schmidt, O. Sliekers, M. Schmid, E. Bock, J. Fuerst, J. G. Kuenen, M. S. Jetten, and M. Strous. New concepts of microbial treatment processes for the nitrogen removal in wastewater. FEMS Microbiology Reviews, 27(4):481–492, Oct. 2003. DOI: 10.1016/s0168-6445(03)00039-1. [185] J. Schmitt and H.-C. Flemming. FTIR-spectroscopy in microbial and material analysis. International Biodeterioration and Biodegradation, 41(1):1–11, Jan. 1998. DOI: 10.1016/s0964-8305(98)80002-4. [186] A. Schmoldt, H. F. Benthe, and G. Haberland. Digitoxin metabolism by rat liver microsomes. Biochemical pharmacology, 24:1639–1641, Sept. 1975. [187] T. Seviour, N. Derlon, M. S. Dueholm, H.-C. Flemming, E. Girbal-Neuhauser, H. Horn, S. Kjelleberg, M. C. van Loosdrecht, T. Lotti, M. F. Malpei, R. Nerenberg, T. R. Neu, E. Paul, H. Yu, and Y. Lin. Extracellular polymeric substances of biofilms: Suffering from an identity crisis. Water Research, 151:1–7, Mar. 2019. DOI: 10.1016/j.watres.2018.11.020. [188] T. Seviour, M. Pijuan, T. Nicholson, J. Keller, and Z. Yuan. Gel-forming exopolysaccharides explain basic differences between structures of aerobic sludge granules and floccular sludges. Water Research, 43(18):4469–4478, Oct. 2009. DOI: 10.1016/j.watres.2009.07.018. [189] G.-P. Sheng, H.-Q. Yu, and X.-Y. Li. Extracellular polymeric substances (EPS) of microbial aggregates in biological wastewater treatment systems: A review. Biotechnology Advances, 28(6):882–894, Nov. 2010. DOI: 10.1016/j. biotechadv.2010.08.001. [190] B. Shivu, S. Seshadri, J. Li, K. A. Oberg, V. N. Uversky, and A. L. Fink. Distinct beta-Sheet Structure in Protein Aggregates Determined by ATR-FTIR Spectroscopy. Biochemistry, 52(31):5176–5183, July 2013. DOI: 10.1021/ bi400625v. [191] K.-Y. Show and D.-J. Lee. Anaerobic Treatment Versus Aerobic Treatment, pages 205–230. Elsevier, Amsterdam, 2017. DOI: 10.1016/ b978-0-444-63665-2.00008-4. [192] K.-Y. Show, Y. Yan, H. Yao, H. Guo, T. Li, D.-Y. Show, J.-S. Chang, and D.-J. Lee. Anaerobic granulation: A review of granulation hypotheses, bioreactor designs and emerging green applications. Bioresource Technology, 300:122751, Mar. 2020. DOI: 10.1016/j.biortech.2020.122751. [193] B. J. Singh, A. Chakraborty, and R. Sehgal. A systematic review of industrial wastewater management: Evaluating challenges and enablers. Journal of Environmental Management, 348:119230, Dec. 2023. DOI: 10.1016/j.jenvman. 2023.119230.
[194] C. M. Singleton, F. Petriglieri, K. Wasmund, M. Nierychlo, Z. Kondrotaite, J. F. Petersen, M. Peces, M. S. Dueholm, M. Wagner, and P. H. Nielsen. The novel genus, candidatus phosphoribacter, previously identified as tetrasphaera, is the dominant polyphosphate accumulating lineage in ebpr wastewater treatment plants worldwide. The ISME Journal, 16(6):1605–1616, Feb. 2022. DOI: 10.1038/s41396-022-01212-z. [195] S. Sivaprakasam, S. Mahadevan, S. Sekar, and S. Rajakumar. Biological treatment of tannery wastewater by using salt-tolerant bacterial strains. Microbial Cell Factories, 7(1), Apr. 2008. DOI: 10.1186/1475-2859-7-15. [196] T. Solov’eva, V. Davydova, I. Krasikova, and I. Yermak. Marine compounds with therapeutic potential in gram-negative sepsis. Marine Drugs, 11(6):2216– 2229, June 2013. DOI: 10.3390/md11062216. [197] W. Song, S. Ding, L. Zhou, N. Li, Y. Zhang, H. Li, J. Ding, and J. Lu. The performance of co-immobilized strains isolated from activated sludge combined with scenedesmus quadricauda to remove nutrients and organics in black odorous water. Bioresource Technology, 345:126571, Feb. 2022. DOI: 10.1016/j.biortech.2021.126571. [198] A. Sonune and R. Ghate. Developments in wastewater treatment methods. Desalination, 167:55–63, 2004. [199] D. T. Sponza. Investigation of extracellular polymer substances (EPS) and physicochemical properties of different activated sludge flocs under steady-state conditions. Enzyme and Microbial Technology, 32(3-4):375–385, Mar. 2003. DOI: 10.1016/s0141-0229(02)00309-5. [200] O. V. Stepanenko, O. V. Stepanenko, I. M. Kuznetsova, V. V. Verkhusha, and K. K. Turoverov. Sensitivity of superfolder gfp to ionic agents. PLoS ONE, 9(10):e110750, Oct. 2014. DOI: 10.1371/journal.pone.0110750. [201] G. Steurs. personal communication. [202] G. Steurs. Advanced NMR Spectroscopy: NMR Data Tables. page 10, 2024. [203] D. Sudmalis, M. Gagliano, R. Pei, K. Grolle, C. Plugge, H. Rijnaarts, G. Zeeman, and H. Temmink. Fast anaerobic sludge granulation at elevated salinity. Water Research, 128:293–303, Jan. 2018. DOI: 10.1016/j.watres.2017.10. 038. [204] D. Sudmalis, S. Millah, M. Gagliano, C. Butre, C. Plugge, H. Rijnaarts, G. Zeeman, and H. Temmink. The potential of osmolytes and their precursors to alleviate osmotic stress of anaerobic granular sludge. Water Research, 147:142–151, Dec. 2018. DOI: 10.1016/j.watres.2018.09.059.
[205] D. Sudmalis, T. Mubita, M. Gagliano, E. Dinis, G. Zeeman, H. Rijnaarts, and H. Temmink. Cation exchange membrane behaviour of extracellular polymeric substances (EPS) in salt adapted granular sludge. Water Research, 178:115855, July 2020. DOI: 10.1016/j.watres.2020.115855. [206] G. P. Szekeres and J. Kneipp. Sers probing of proteins in gold nanoparticle agglomerates. Frontiers in Chemistry, 7, Jan. 2019. DOI: 10.3389/fchem. 2019.00030. [207] M. G. Tecson, D. H. Camacho, V. D. Ebajo, and L. V. Abad. Structural investigation of degradation products of irradiated kappa-carrageenan. Radiation Physics and Chemistry, 194:110015, May 2022. DOI: 10.1016/j.radphyschem. 2022.110015. [208] T. Thrimawithana, S. Young, D. Dunstan, and R. Alany. Texture and rheological characterization of kappa and iota carrageenan in the presence of counter ions. Carbohydrate Polymers, 82(1):69–77, Aug. 2010. DOI: 10.1016/j.carbpol.2010.04.024. [209] X. Tian, Z. Shen, Z. Han, and Y. Zhou. The effect of extracellular polymeric substances on exogenous highly toxic compounds in biological wastewater treatment: An overview. Bioresource Technology Reports, 5:28–42, Feb. 2019. DOI: 10.1016/j.biteb.2018.11.009. [210] United States Environmental Protection Agency. Effluent guidelines, 2024. https://www.epa.gov/eg, [Accessed on 11/11/2024]. [211] R. Van den Broeck. Activated sludge mixed liquor characteristics versus membrane fouling in MBRs. PhD thesis, KULeuven, 2011. [212] B. Van der Bruggen. Waste Water Treatment Technology Part I: Legislation in Flanders and Primary Waste Water Technology. Acco Leuven, 2018. [213] J. Van Dierdonck. Impact of Influent Characteristics on Activated Sludge Bioflocculation in Lab-scale Systems. PhD thesis, KULeuven, 2013. [214] K. van Leeuwen, E. de Vries, S. Koop, and K. Roest. The energy and raw materials factory: Role and potential contribution to the circular economy of the netherlands. Environmental Management, 61(5):786–795, Jan. 2018. DOI: 10.1007/s00267-018-0995-8. [215] J. B. van Lier, F. P. van der Zee, C. T. M. J. Frijters, and M. E. Ersahin. Celebrating 40 years anaerobic sludge bed reactors for industrial wastewater treatment. Reviews in Environmental Science and Biotechnology, 14(4):681–702, Aug. 2015. DOI: 10.1007/s11157-015-9375-5. [216] Vlaamse Milieu Maatschappij. Wet op de bescherming van de oppervlaktewateren tegen verontreiniging, 2024. https://www.vmm.be/wetgeving/wet-op-de-bescherming-van-de-oppervlaktewa… [Accessed on 11/11/2024]
[217] I. Vyrides, H. Santos, A. Mingote, M. J. Ray, and D. C. Stuckey. Are compatible solutes compatible with biological treatment of saline wastewater? batch and continuous studies using submerged anaerobic membrane bioreactors (sambrs). Environmental Science and Technology, 44(19):7437–7442, Sept. 2010. DOI: 10.1021/es903981k. [218] M. Wagner, N. P. Ivleva, C. Haisch, R. Niessner, and H. Horn. Combined use of confocal laser scanning microscopy (CLSM) and Raman microscopy (RM): Investigations on EPS - Matrix. Water Research, 43(1):63–76, Jan. 2009. DOI: 10.1016/j.watres.2008.10.034. [219] D. Wang, N. Dong, S. Hui, and Y. Niu. Analysis of urea pyrolysis products in 132.5-190degreesc. Energy Procedia, 158:2170–2175, Feb. 2019. DOI: 10.1016/ j.egypro.2019.01.616. [220] J. Wang and H. Chen. Catalytic ozonation for water and wastewater treatment: Recent advances and perspective. Science of The Total Environment, 704:135249, Feb. 2020. DOI: 10.1016/j.scitotenv.2019.135249. [221] Z. Wang, Z. Wu, and S. Tang. Extracellular polymeric substances (EPS) properties and their effects on membrane fouling in a submerged membrane bioreactor. Water Research, 43(9):2504–2512, May 2009. DOI: 10.1016/j. watres.2009.02.026. [222] C. Wei, F. Zhang, Y. Hu, C. Feng, and H. Wu. Ozonation in water treatment: the generation, basic properties of ozone and its practical application. Reviews in Chemical Engineering, 33(1):49–89, July 2016. DOI: 10.1515/revce-2016-0008. [223] Y. Wei, R. T. Van Houten, A. R. Borger, D. H. Eikelboom, and Y. Fan. Minimization of excess sludge production for biological wastewater treatment. Water Research, 37(18):4453–4467, Nov. 2003. DOI: 10.1016/s0043-1354(03) 00441-X. [224] D. T. Welsh. Ecological significance of compatible solute accumulation by microorganisms: from single cells to global climate. FEMS Microbiology Reviews, 24(3):263–290, July 2000. DOI: 10.1111/j.1574-6976.2000.tb00542.x. [225] M. K. Winkler, R. Kleerebezem, W. O. Khunjar, B. de Bruin, and M. C. van Loosdrecht. Evaluating the solid retention time of bacteria in flocculent and granular sludge. Water Research, 46(16):4973–4980, Oct. 2012. DOI: 10.1016/j.watres.2012.06.027. [226] Y. Xiao and D. J. Roberts. A review of anaerobic treatment of saline wastewater. Environmental Technology, 31(8):1025–1043, jul 2010. DOI: 10.1080/09593331003734202.
227] W. Xing, M. Yin, Q. Lv, Y. Hu, C. Liu, and J. Zhang. Oxygen Solubility, Diffusion Coefficient, and Solution Viscosity, pages 1–31. Elsevier, Amsterdam, 2014. DOI: 10.1016/b978-0-444-63278-4.00001-x. [228] H.-Y.-Y. Yao, J.-Q. Wang, J.-Y. Yin, S.-P. Nie, and M.-Y. Xie. A review of NMR analysis in polysaccharide structure and conformation: Progress, challenge and perspective. Food Research International, 143:110290, May 2021. DOI: 10.1016/j.foodres.2021.110290. [229] D. P. Zagklis and G. Bampos. Tertiary Wastewater Treatment Technologies: A Review of Technical, Economic, and Life Cycle Aspects. Processes, 10(11):2304, nov 2022. DOI: 10.3390/pr10112304. [230] S. A. Zahra, R. Persiani, M. K. Dueholm, M. van Loosdrecht, P. H. Nielsen, T. W. Seviour, and Y. Lin. Rethinking characterization, application, and importance of extracellular polymeric substances in water technologies. Current Opinion in Biotechnology, 89:103192, Oct. 2024. DOI: 10.1016/j.copbio. 2024.103192. [231] N. S. Zaidi, K. Muda, and M. Sillanpaa. Effects of extra-cellular polymeric substances towards physical properties of biomass under magnetic field exposure. International Journal of Environmental Science and Technology, 16(7):3801– 3808, July 2018. DOI: 10.1007/s13762-018-1897-2. [232] W. Zeng, F. Li, C. Wu, R. Yu, X. Wu, L. Shen, Y. Liu, G. Qiu, and J. Li. Role of extracellular polymeric substance (EPS) in toxicity response of soil bacteria Bacillus sp. S3 to multiple heavy metals. Bioprocess and Biosystems Engineering, 43(1):153–167, Sept. 2019. DOI: 10.1007/s00449-019-02213-7. [233] L. Zhang, X. Lin, Z. Zhang, G.-H. Chen, and F. Jiang. Elemental sulfur as an electron acceptor for organic matter removal in a new high-rate anaerobic biological wastewater treatment process. Chemical Engineering Journal, 331:16– 22, Jan. 2018. DOI: 10.1016/j.cej.2017.08.055. [234] S. Zhang, S. Wakai, N. Sasakura, H. Tsutsumi, Y. Hata, C. Ogino, and A. Kondo. Pyruvate metabolism redirection for biological production of commodity chemicals in aerobic fungus aspergillus oryzae. Metabolic Engineering, 61:225–237, Sept. 2020. DOI: 10.1016/j.ymben.2020.06.010. [235] J. Zheng, C. Su, J. Zhou, L. Xu, Y. Qian, and H. Chen. Effects and mechanisms of ultraviolet, chlorination, and ozone disinfection on antibiotic resistance genes in secondary effluents of municipal wastewater treatment plants. Chemical Engineering Journal, 317:309–316, June 2017. DOI: 10.1016/j.cej.2017.02. 076. [236] Y. N. Zhou, J. Li, J. Li, and S. Wei. Dewaterability of aerobic granular sludge. Applied Mechanics and Materials, 90-93:2944–2948, Sept. 2011. DOI: 10.4028/www.scientific.net/amm.90-93.2944
[237] L. Zhu, H.-y. Qi, M.-l. Lv, Y. Kong, Y.-w. Yu, and X.-y. Xu. Component analysis of extracellular polymeric substances (EPS) during aerobic sludge granulation using FTIR and 3D-EEM technologies. Bioresource Technology, 124:455–459, Nov. 2012. DOI: 10.1016/j.biortech.2012.08.059. [238] J. Zlopasa, E. Koenders, and S. Picken. Using bio-based polymers for curing cement-based materials. In AMS 14 Proceedings of the Int. Conference on Ageing of Materials and Structures, pages 220–226, 2014. [239] V. Zorraquino, D. Toubiana, D. Yan, and E. Blumwald. Draft Genome Sequence of the Nitrogen-Fixing Endophyte Azoarcus communis SWub3. Microbiology Resource Announcements, 7(13), Oct. 2018. DOI: 10.1128/mra.01080-18. [240] E. Zuriaga-Agusti, A. Bes-Pia, J. Mendoza-Roca, and J. Alonso-Molina. Influence of extraction methods on proteins and carbohydrates analysis from MBR activated sludge flocs in view of improving EPS determination. Separation and Purification Technology, 112:1–10, July 2013. DOI: 10.1016/j.seppur.2013.03.048.