Understanding the role of Tetrasphaera in enhanced biological - PowerPoint PPT Presentation

Understanding the role of Tetrasphaera in enhanced biological phosphorus removal N. Rey, M. Badia, A. Guisasola, J. A. Baeza Departament d’Enginyeria Química Biològica i Ambiental, Escola d’Enginyeria, Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona 13 th IWA Specialized Conference on Small Water and Wastewater Systems 14-16 September, 2016 ATHENS, GREECE

1 Introduction EBPR is an economical, efficient and sustainable way to remove EBPR Alternating anaerobic and aerobic conditions phosphorus from the wastewater PAOs Uptake and storage of poly-phosphate PHA Take up simple carbon sources Candidatus Accumulibacter phosphatis Most well-known and studied PAO Genus Tetrasphaera Full-scale EBPR plants Other putative PAOs Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

1 Introduction Volatile fatty acids Lab-scale COD consumed in the anaerobic reactor/phase Microbiological techniques Enrichment of Accumulibacter (PAOs) Low concentrations of volatile fatty acids Full-scale Amino acids Tetrasphaera proliferation Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

2 Objective  To gain knowledge on the new PAO genus Tetrasphaera by obtaining an enriched culture at lab-scale  Two reactor configurations:  Sequencing batch reactor  Continuous pilot plant system with A 2 /O configuration Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: SBR operation V = 10L Synthetic influent: 10 mg P-PO 4 -3 /L, 44-113 mg COD/L Aspartate + Glutamate Glutamate 40 Phosphate concentration (mg P·L -1 ) G+A G+A G G+A Single batch experiments 44 mg O 2 /L 113 71 mg O 2 /L 71 mg O 2 /L mg O 2 /L Aspartate 30 20 N2 increase 10 Initial time End of anaerobic phase End of aerobic phase 0 0 20 40 60 80 100 Time (days) Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: SBR operation V = 10L Synthetic influent: 10 mg P-PO 4 -3 /L, 44-113 mg COD/L Aspartate + Glutamate 40 Phosphate concentration (mg P·L -1 ) Excess of carbon source G+A G+A G G+A 44 mg O 2 /L 113 71 mg O 2 /L 71 mg O 2 /L mg O 2 /L 30 Glutamate leakage to the aerobic phase 20 N2 increase System failure 10 Initial time 0 End of anaerobic phase 0 20 40 60 80 100 End of aerobic phase Time (days) Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: A 2 /O operation V = 146L Synthetic influent: 10 mg P-PO 4 -3 /L, 50 mg N-NH 4 + /L and 400 mg COD/L Glutamate Glutamate as carbon and nitrogen source P-removal efficiency>95% ) -1 100 100 Phosphate concentration (mg·L Batch deteriorated P-removal efficiency (%) 80 80 glutamate 60 60 - input Excesive NO 3 40 40 in the anaerobic reactor 20 20 P-PO4-3 Anaerobic (mg/L) 0 0 P-PO4-3 Effluent (mg/L) 0 100 200 300 400 P-removal efficiency (%) Time (days) Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Batch experiments with the SBR sludge PHA & Glycogen quantification Anaerobic PHA and glycogen (mmol C/g VSS) Aerobic 40 5 PHA: 0.69 mmol C/gVSS Phosphate (mg P/L) 4 30 Glycogen: TOC (mg C/L) 0.47 mmol C/gVSS 3 20 2 PAO-enriched cultures: 10 PHA production: 1 2.67 mmol C/gVSS Glycogen consumption: 0 0 0 50 100 150 200 250 300 350 1.30 mmol C/gVSS Time (min) P (mg/L) TOC (mg/L) Glycogen (mmol C/g VSS) PHA (mmol C/g VSS) Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Batch experiment with the A 2 /O sludge PHA & Glycogen quantification Anaerobic Aerobic 100 2.5 PHA and glycogen (mmol C/mg VSS) Phosphate (mg P/L) and COD (mg/L) PHA: 0.62 mmol C/gVSS 80 2.0 Glycogen: 60 1.5 0.06 mmol C/gVSS 40 1.0 PAO-enriched cultures: PHA production: 20 0.5 2.67 mmol C/gVSS Glycogen consumption: 0 0.0 1.30 mmol C/gVSS 0 50 100 150 200 250 300 350 Time (min) P (mg/L) COD (mg/L) Glycogen (mmol C/g SSV) PHA (mmol C/g SSV) Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Comparison between configurations Anaerobic PHA and glycogen production mmol C/g VSS SBR A 2 /O COD consumed 3.436 8.87 PHA 0.692 0.621 Glycogen 0.476 0.066 Total=PHA + Glycogen 1.168 0.687 Carbon recovery ratio 0.34 0.08 Other storage routes should be studied Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Literature comparison P rel /C upt PHA prod /C upt Glyc prod /C upt Study (mol P/mol C) (mol C/mol C) (mol C/mol C) Enriched Kapagiannidis 0.64 1.10 Consumption PAO et al. (2013) (-0.41) Tayà et al. 0.34 1.47 Consumption (2013) (-0.49) This study SBR 0.27 0.20 0.14 A 2 /O 0.21 0.07 0.01 Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Bacterial community assessment All bacteria Specific bacteria SBR This image cannot currently be displayed. This image cannot currently be displayed. 30% 13% Uncultured Tetrasphaera Tetrasphaera Tetrasphaera Clone ASM31 Clade II Clone ASM47 Tetraspahera 21% 31% PAO GAO Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Bacterial community assessment All bacteria Specific bacteria A 2 /O pilot plant 27% 39% Uncultured Tetrasphaera Tetrasphaera Tetrasphaera Clone ASM31 Clade II Clone ASM47 Tetraspahera 26% 1% PAO GAO Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Bacterial community assessment PAOMix GAOMix Tetrasphaera SBR 36 ± 1% 21 ± 1% 43 ± 9% A 2 /O plant 26 ± 4% 1 ± 1% 66 ± 5% Why do we detect the presence of PAO and GAO if we fed the reactor with glutamate for more than 400 days? Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

3 Results: Bacterial community assessment Anaerobic fermentation routes for glutamate Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

4 Further work We are still operating the A 2 /O pilot plant with glutamate We are working with this enriched- tetrasphaera culture in order to better understanding this new PAO genus We will perform batch assays with different carbon sources and different electron acceptors We are waiting for the pyrosequencing results Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

5 Conclusions • Successful enrichment of sludge in Tetrasphaera using glutamate as sole carbon source was obtained for the first time. • Better results and more stability was achieved with continuous pilot plant with respect to SBR. • Fermentation products of glutamate did not allow to obtain a highly Tetrasphaera -enriched culture. • The increase of PHA and glycogen during the anaerobic phase only accounted a small percentage of the carbon source consumed. • Other storage routes should be studied to identify the fate of the carbon source stored under anaerobic conditions. Understanding the role of Tetrasphaera in EBPR – N. Rey et al. SWWS 2016

Ackno knowledgments ledgments This image cannot currently be displayed. CTQ2014-60495-R with funds from the Fondo Europeo de Desarrollo Regional (FEDER)

Thank you for your attention! talia Rey talia Rey Marina Badia Marina Badia Albert Guisasola Albert Guisasola Juan Antonio Baeza uan Antonio Baeza

Understanding the role of Tetrasphaera in enhanced biological phosphorus removal N. Rey, M. Badia, A. Guisasola, J. A. Baeza Departament d’Enginyeria Química Biològica i Ambiental, Escola d’Enginyeria, Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona