Coagulation in combination with anaerobic digestion for enhancement of resource recovery from faecal sludge.

Poorly managed faecal sludge (FS) poses significant challenges to public health and the environment. Anaerobic digestion (AD) of FS provides an effective method for energy recovery while reducing FS associated threats. Recognizing the critical role of the dewatering process before AD, this study investigates the synergistic application of chemical coagulation and mesophilic AD for synthetic FS treatment. FeCl3, AlCl3, Fe2(SO4)3, poly ferric sulfate (PFS) and poly aluminium ferric chloride (PAFC) were utilized at varying dosages to examine their impact on FS properties and subsequent biogas production from the dewatered FS. It was found that coagulation enhances sedimentation efficiencies and dewaterability through mechanisms such as charge neutralization, charge patching and bridging, thereby improving the FS feasibility for AD. Notably, polymer coagulant PFS showed good performance in balancing pollutant removal and methane recovery, contributing to facilitating the hydrolysis and acidogenesis microorganisms involved in the AD process. Optimal dosage was identified at 150 mg/g TS (1.7 g/L FS), achieving prominent removal efficiencies for total COD (67%), turbidity (85%), and total phosphorus (60%), while simultaneously enhancing AD performance with specific CH4 production reaching 517 ml CH4/g VS or 24.8 ml CH4/g AD wet feedstock compared to 309 ml CH4/g VS or 2.7 ml CH4/g AD wet feedstock in untreated FS.

Total value wall: Full scale demonstration of a green wall for grey water treatment and recycling.

Greywater treatment and reuse for non-potable purposes in urban areas has become a widely researched topic to reduce the burden on fresh water resources. This study reports on the use of a green wall for treating grey water and reusing the effluent for toilet flushing, called Total Value Wall (TVW). Initially, the effectiveness of (mixtures of) different substrates, i.e. lava, lightweight expanded clay aggregates, organic soil and biochar was investigated by means of column tests. All substrates were first examined for hydraulic characteristics and later on the columns were fed with synthetic grey wastewater and followed up in terms of removal efficiency of COD and detergents. The mixture consisting of lava (50%), organic soil (25%) and biochar (25%) proved to be optimal both in terms of percolation rates and removal efficiencies, and was thus selected for the full-scale system. The full-scale TVW of 14.4 m2 was installed at a terraced house in Ghent (Belgium), and was loaded with grey water at 100 L per day. Influent and effluent quality were routinely monitored by grab sampling, water savings were monitored by means of flow meters, and electricity consumption was also accounted for. The TVW was further equipped with sensors that measure temperature, Particulate Matter (PM10) and CO2 in the air. The full-scale system obtained effluent concentrations of 13 mg.L-1 TSS, 91 mg.L-1 COD and 5 mg.L-1 BOD5. Ammonium and total coliforms were removed with removal rates of 97% and 99% (2 log units) respectively. However, an increase in effluent concentration of nitrate and phosphate was observed due to leaching from the selected substrate. Available data from the temperature sensors have clearly demonstrated the additional benefit of the TVW as an insulating layer, keeping the heat outside on warmer days, and keeping the heat inside on colder days. Overall, this study demonstrated that the TVW is a sustainable system for greywater treatment and reuse.

PARAFAC model as an innovative tool for monitoring natural organic matter removal in water treatment plants.

The increase of fluorescent natural organic matter (fNOM) fractions during drinking water treatment might lead to an increased coagulant dose and filter clogging, and can be a precursor for disinfection by-products. Consequently, efficient fNOM removal is essential, for which characterisation of fNOM fractions is crucial. This study aims to develop a robust monitoring tool for assessing fNOM fractions across water treatment processes. To achieve this, water samples were collected from six South African water treatment plants (WTPs) during winter and summer, and two plants in Belgium during spring. The removal of fNOM was monitored by assessing fluorescence excitation-emission matrices datasets using parallel factor analysis. The removal of fNOM during summer for South African WTPs was in the range 69-85%, and decreased to 42-64% in winter. In Belgian WTPs, fNOM removal was in the range 74-78%. Principal component analysis revealed a positive correlation between total fluorescence and total organic carbon (TOC). However, TOC had an insignificant contribution to the factors affecting fNOM removal. Overall, the study demonstrated the appearance of fNOM in the final chlorinated water, indicating that fNOM requires a customised monitoring technique.

Anaerobic treatment of blended sugar industry and ethanol distillery wastewater through biphasic high rate reactor.

This study aimed to investigate the physicochemical properties of sugar industry and ethanol distillery wastewater and the treatment of the blended wastewater through a two-stage anaerobic reactor. For this treatment, different initial chemical oxygen demand (COD) concentrations (5-20 g/L) and hydraulic retention times (HRTs) (2-10 days) were applied. The sugar industry effluent characteristics obtained in terms of organic matter (mg/L) were as follows: 5 days biochemical oxygen demand (BOD5): 654.5-1,968; COD: 1,100-2,148.9; total solids (TS): 2,467-4,012 mg/L; and pH: 6.93-8.43. The ethanol distillery spent wash strengths obtained were: BOD5: 27,600-42,921 mg/L; COD: 126,000-167,534 mg/L; TS: 140,160-170,000 mg/L; and pH: 3.9-4.2. Maximum COD removal of 65% was obtained at optimum condition (initial COD concentration of 10 g/L and HRT of 10 days), and maximum color removal of 79% was recorded under similar treatment conditions. Hence, the performance of the two-stage anaerobic reactor for simultaneous removal of COD and color from high-strength blended wastewater is promising for scaling up in order to mitigate environmental problems of untreated effluent discharge.

Surrogate-Based Correlation Models in View of Real-Time Control of Ozonation of Secondary Treated Municipal Wastewater-Model Development and Dynamic Validation.

New robust correlation models for real-time monitoring and control of trace organic contaminant (TrOC) removal by ozonation are presented, based on UVA254 and fluorescence surrogates, and developed considering kinetic information. The abatement patterns of TrOCs had inflected shapes, controlled by the reactivity of TrOCs toward ozone and HO• radicals. These novel and generic correlation models will be of importance for WRRF operators to reduce operational costs and minimize byproduct formation. Both UVA254 and fluorescence surrogates could be used to control ΔTrOC, although fluorescence measurements indicated a slightly better reproducibility and an enlarged control range. The generic framework was validated for several WRRFs and correlations for any compound with known kinetic information could be developed solely using the second order reaction rate constant with ozone (kO3). Two distinct reaction phases were defined for which separate linear correlations were obtained. The first was mainly ozone controlled, while the second phase was more related to HO• reactions. Furthermore, parallel factor analysis of the fluorescence spectra enabled monitoring of multiple types of organic matter with different O3 and HO• reactivity. This knowledge is of value for kinetic modeling frameworks and for achieving a better understanding of the occurring changes of organic matter during ozonation.

Removal of organic matter and ammonium from landfill leachate through different scenarios: Operational cost evaluation in a full-scale case study of a Flemish landfill.

Several scenarios are available to landfilling facilities to effectively treat leachate at the lowest possible cost. In this study, the performance of various leachate treatment sequences to remove COD and nitrogen from a leachate stream and the associated cost are presented. The results show that, to achieve 100% nitrogen removal, autotrophic nitrogen removal (ANR) or a combination of ANR and nitrification - denitrification (N-dN) is more cost effective than using only the N-dN process (0.58 €/m3) without changing the leachate polishing costs associated with granular activated carbon (GAC). Treatment of N-dN effluent by ozonation or coagulation led to the reduction of the COD concentration by 10% and 59% respectively before GAC adsorption. This reduced GAC costs and subsequently reduced the overall treatment costs by 7% (ozonation) and 22% (coagulation). On the contrary, using Fenton oxidation to reduce the COD concentration of N-dN effluent by 63% increased the overall leachate treatment costs by 3%. Leachate treatment sequences employing ANR for nitrogen removal followed by ozonation or Fenton or coagulation for COD removal and final polishing with GAC are on average 33% cheaper than a sequence with N-dN + GAC only. When ANR is the preceding step and GAC the final step, choice of AOP i.e., ozonation or Fenton did not affect the total treatment costs which amounted to 1.43 (ozonation) and 1.42 €/m3 (Fenton). In all the investigated leachate treatment trains, the sequence with ANR + coagulation + GAC is the most cost effective at 0.94 €/m3.

Enhanced treatment of secondary municipal wastewater effluent: comparing (biological) filtration and ozonation in view of micropollutant removal, unselective effluent toxicity, and the potential for real-time control.

Ozonation and three (biological) filtration techniques (trickling filtration (TF), slow sand filtration (SSF) and biological activated carbon (BAC) filtration) have been evaluated in different combinations as tertiary treatment for municipal wastewater effluent. The removal of 18 multi-class pharmaceuticals, as model trace organic contaminants (TrOCs), has been studied. (Biological) activated carbon filtration could reduce the amount of TrOCs significantly (>99%) but is cost-intensive for full-scale applications. Filtration techniques mainly depending on biodegradation mechanisms (TF and SSF) are found to be inefficient for TrOCs removal as a stand alone technique. Ozonation resulted in 90% removal of the total amount of quantified TrOCs, but a post-ozonation step is needed to cope with an increased unselective toxicity. SSF following ozonation showed to be the only technique able to reduce the unselective toxicity to the same level as before ozonation. In view of process control, innovative correlation models developed for the monitoring and control of TrOC removal during ozonation, are verified for their applicability during ozonation in combination with TF, SSF or BAC. Particularly for the poorly ozone reactive TrOCs, statistically significant models were obtained that correlate TrOC removal and reduction in UVA254 as an online measured surrogate parameter.

Methane oxidation in a biofilter (Part 2): A lab-scale experiment for model calibration.

In this study an experimental study on a biological methane oxidation column presented with the aim to calibrate a mathematical model developed in an earlier study. The column was designed to reproduce at lab-scale a real biofilter trying to consider the more probable landfill boundary conditions. Although the methane oxidation efficiency in the column was lower than the expected (around 35%), an appropriate model implementation showed an acceptable agreement between the outcomes data of the model simulation and the experimental data (with Theil's Inequality Coefficient value of 0.08). A calibrated model allows a better management of the biofilter performance in terms of methane oxidation.

Methane oxidation in a biofilter (Part 1): Development of a mathematical model for designing and optimization.

The aim of this work is the evaluation of the efficiency of such a biofilter, through the application of a mathematical model which describes the biological oxidation process. This mathematical model is able to predict the efficiency of the system under varying operating conditions. Literature data have been used in order to build the model. The factors that mostly affect the process and which actually regulate the entire process have been highlighted in this work. Specifically, it was found that temperature, flow and methane concentration are the most important parameters that influence the system. The results obtained from the mathematical model showed also that the biofilter system is simple to implement and manage and allows the achievement of high efficiency of methane oxidation. In the optimal conditions for temperature (between 20-30°C), residence time (between 0.7-0.8 h) and methane molar fraction (between 20-25%) the efficiency of methane oxidation could be around 50%.

TREATMENT OF LANDFILL LEACHATE BY COUPLING COAGULATION-FLOCCULATION OR OZONATION TO GRANULAR ACTIVATED CARBON ADSORPTION.

A major concern for landfilling facilities is the treatment of their leachate. To optimize organic matter removal from this leachate, the combination of two or more techniques is preferred in order to meet stringent effluent standards. In our study, coagulation-flocculation and ozonation are compared as pre- treatment steps for stabilized landfill leachate prior to granular activated carbon (GAC) adsorption. The efficiency of the pre treatment techniques is evaluated using COD and UVA254 measurements. For coagulation- flocculation, different chemicals are compared and optimal dosages are determined. After this, iron (III) chloride is selected for subsequent adsorption studies due to its high percentage of COD and UVA254 removal and good sludge settle-ability. Our finding show that ozonation as a single treatment is effective in reducing COD in landfill leachate by 66% compared to coagulation flocculation (33%). Meanwhile, coagulation performs better in UVA254 reduction than ozonation. Subsequent GAC adsorption of ozonated effluent, coagulated effluent and untreated leachate resulted in 77%, 53% and 8% total COD removal respectively (after 6 bed volumes). The effect of the pre-treatment techniques on GAC adsorption properties is evaluated experimentally and mathematically using Thomas and Yoon-Nelson models. Mathematical modelling of the experimental GAC adsorption data shows that ozonation increases the adsorption capacity and break through time with a factor of 2.5 compared to coagulation-flocculation.

Performance and kinetic process analysis of an Anammox reactor in view of application for landfill leachate treatment.

Anammox has shown its promise and low cost for removing nitrogen from high strength wastewater such as landfill leachate. A reactor was inoculated with nitrification-denitrification sludge originating from a landfill leachate treating waste water treatment plant. During the operation, the sludge gradually converted into red Anammox granular sludge with high and stable Anammox activity. At a maximal nitrogen loading rate of 0.6 g N l(-1) d(-1), the reactor presented ammonium and nitrite removal efficiencies of above 90%. In addition, a modified Stover-Kincannon model was applied to simulate and assess the performance of the Anammox reactor. The Stover-Kincannon model was appropriate for the description of the nitrogen removal in the reactor with the high regression coefficient values (R2 = 0.946) and low Theil's inequality coefficient (TIC) values (TIC < 0.3). The model results showed that the maximal N loading rate of the reactor should be 3.69 g N l(-1) d(-).

Lab-scale evaluation of Microalgal-Bacterial granular sludge as a sustainable alternative for brewery wastewater treatment.

Microalgal-bacterial granular sludge (MBGS) could offer a sustainable alternative to traditional aerobic methods in brewery wastewater (BWW) treatment. This study compared MBGS with conventional activated sludge (AS) in treating real BWW and highlighted its advantages and challenges. MBGS achieved comparable chemical oxygen demand removal efficiency (93%) compared to AS (89%). Additionally, MBGS exhibited higher phosphate removal capabilities than AS. Extra nitrogen was added to influent to balance C/N ratio of BWW. MBGS was robust in handling C/N ratio fluctuations with an 82% total nitrogen removal efficiency. Metagenomic analysis further indicated that most of the genes involved in carbon, nitrogen and phosphorus metabolism were up-regulated in MBGS compared to AS. Despite changes in the microbial community and settling ability due to high starch and sugar content in BWW, MBGS demonstrated high efficiency and sustainability. Further research should optimize MBGS operation strategies to fully realize its potential for sustainable BWW treatment.

Hybrid packed bed bioreactor using combined biodegradation and ozonation to enhance nitrogen and micropollutants removal from landfill leachate.

Landfill leachate contains ammonium and micropollutants. Ammonium can be biologically removed but bio-recalcitrant micropollutants removal requires post-treatment like ozonation. This study developed an expanded clay aggregates packed biofilm column (EBC) and demonstrated its feasibility of coupling biodegradation and ozonation (CBAO) to simultaneously remove nitrogen and bio-recalcitrant micropollutants. The first 60 days only had biodegradation process to start the bioreactor. 51 % nitrogen was biologically removed but the removal of micropollutant carbamazepine (CBZ) was only 30 %. From 61 d to 150 d, both biodegradation and ozonation were performed in the EBC. After 48 h-biodegradation, ozone gas was introduced and bubbling through EBC for 30 min to further remove residual micropollutants. At 0.4 gO3/gCOD, CBZ were completely removed. The average nitrogen removal efficiency (85 %) was increased by 34 % because the increased abundance of nitrifying and denitrifying bacteria in EBC. This study confirmed the promising potential of the CBAO process for treating landfill leachte.

Improving the ozone-activated peroxymonosulfate process for removal of trace organic contaminants in real waters through implementation of an optimized sequential ozone dosing strategy.

The ozone-activated peroxymonosulfate process (O3/PMS) has received increasing attention for the removal of trace organic contaminants (e.g. pesticides and pharmaceuticals) from water bodies. However, the ozone dosing strategy has not yet been properly investigated, especially in real water matrices. Typically, one-step dosing is applied in literature. Nevertheless, optimal dosing is an important step for improving the process. This study investigates the effect of sequential ozone dosing on the PMS activation, atrazine (ATZ) removal, residual ozone concentration and radical exposure, and compares the results to those of a one-step ozone dosing approach. Experiments were performed in three water matrices with a different (in)organic content, i.e. secondary effluent, surface water and groundwater. In all matrices, the highest PMS activation was reached when applying three sequential ozone doses (3 × 5 mg O3/L). This resulted in a 3 times higher ATZ removal efficiency (up to 46 %) in secondary effluent compared to that obtained with a one-step ozone dosing (15 mg O3/L). In surface water and groundwater, similar ATZ removal (>90 %) was observed among the different ozone dosing strategies. However, the sulfate radical (SO4●-) exposure increased after each ozone addition. After three ozone additions of 5 mg/L, SO4●- contributed for 9 %, 26 % and 54 % to ATZ removal in respectively secondary effluent, surface water and groundwater. This high SO4●- contribution compared to ●OH contribution is an advantage as the selectivity of SO4●- gives rise to less radical scavenging by bulk organic matter and thus increases the (cost-)effectiveness of the O3/PMS process.

Removal of organic micropollutants from water by adsorption on thermo-plasma expanded graphite encapsulated into calcium alginate.

Nowadays, public concern is focused on the degradation of water quality. For this reason, the development of innovative technologies for water treatment in view of (micro)pollutant removal is important. Indeed, organic (micro)pollutants, such as pharmaceuticals, herbicides, pesticides and plasticizers at concentration levels of µg L-1 or even ng L-1 are hardly removed during conventional wastewater treatment. In view of this, thermo-plasma expanded graphite, a light-weight innovative material in the form of a powder, was encapsulated into calcium alginate to obtain a granular form useful as filtration and adsorption material for removal of different pollutants. The produced material was used to remove atrazine, bisphenol-A, 17-α-ethinylestradiol and carbamazepine (at concentration levels of 125, 250 and 500 µg L-1) by top-down filtration. The effect of flow rate, bed depth and adsorbent composition was evaluated based on breakthrough curves. The experimental data was analysed with the Adams-Bohart model in view of scale-up. Under optimal conditions, removal and adsorption capacity of respectively about 21%, 21%, 38%,42%, 43 µg g-1, 44 µg g-1, 37 µg g-1 and 87 µg g-1 were obtained for atrazine, bisphenol, 17-α ethinylestradiol and carbamazepine when using 0.12 g of thermo-plasma expanded graphite to treat 200 mL at 500 µg L-1 (for each compound) of solution obtaining at contact time of 20 min. The granular form of TPEG obtained (GTPEG) by entrapping in calcium alginate results to have a good adsorbent property for the removal of carbamazepine, atrazine, bisphenol A and 17-α ethinylestradiol from water at concentration levels between 250 and 500 µg L-1. Promising results confirm the adsorbent properties of TPEG and push-up us to investigate on its application and improve of its performance by evaluating different entrapping materials. Supplementary Information: The online version contains supplementary material available at 10.1007/s40201-023-00876-9.

Enhanced removal of refractory humic- and fulvic-like organics from biotreated landfill leachate by ozonation in packed bubble columns.

Biotreated landfill leachate contains much refractory organics such as humic and fulvic acids, which can be degraded by O3. However, the low O3 mass transfer and high energy cost limit its wide application in landfill leachate treatment. Previous studies proved that packed bubble columns could enhance the O3 mass transfer and increase the synthetic humic acids wastewater degradation, but the performance of packed bubble columns in real wastewater treatment has not been investigated. Therefore, this study aims to evaluate the feasibility of application of packed bubble column in the real biotreated landfill leachates treatment and provide insights into the transformation of organic matters in leachates during ozonation. Packed bubble columns with lava rocks or metal pall rings (LBC or MBC) were applied and compared with a non-packed bubble column (BC). At an applied O3 dose of 8.35 mg/(Lwater sample min), the initial COD (400 mg/L) was only removed for 26% in BC and 32% in MBC while this was 46% in LBC, indicating LBC has the best performance. GC-MS analysis shows that raw biotreated leachate contains potential endocrine disruptors such as di(2-ethylhexyl) phthalate (DEHP). 61% of DEHP was removed in LBC and the least intermediate oxidation products from humic- and fulvic-like organics was detected in LBC. The highest O3 utilization efficiency (89%) and hydroxyl radical (OH) exposure rate (3.0 × 10-10 M s) were observed in LBC with lowest energy consumption (EEO) for COD removal of 18 kWh/m3. The enhanced ozonation efficiency in LBC and MBC was attributed to the improved O3 mass transfer. Besides, LBC had additional adsorptive and catalytic activity that promoted the decomposition of O3 to generate OH. This study demonstrates that a packed bubble column increases removal and decreases energy use when treating landfill leachate, thus promoting the application of ozonation.

Life cycle assessment of microalgae systems for wastewater treatment and bioproducts recovery: Natural pigments, biofertilizer and biogas.

The aim of this study was to assess the potential environmental impacts associated with microalgae systems for wastewater treatment and bioproducts recovery. In this sense, a Life Cycle Assessment was carried out evaluating two systems treating i) urban wastewater and ii) industrial wastewater (from a food industry), with the recovery of bioproducts (i.e. natural pigments and biofertilizer) and bioenergy (i.e. biogas). Additionally, both alternatives were compared to iii) a conventional system using a standard growth medium for microalgae cultivation in order to show the potential benefits of using wastewater compared to typical cultivation approaches. The results indicated that the system treating industrial wastewater with unialgal culture had lower environmental impacts than the system treating urban wastewater with mixed cultures. Bioproducts recovery from microalgae wastewater treatment systems can reduce the environmental impacts up to 5 times compared to a conventional system using a standard growth medium. This was mainly due to the lower chemicals consumption for microalgae cultivation. Food-industry effluent showed to be the most promising scenario for bioproducts recovery from microalgae treating wastewater, because of its better quality compared to urban wastewater which also allows the cultivation of a single microalgae species. In conclusion, microalgae wastewater treatment systems are a promising solution not only for wastewater treatment but also to boost the circular bioeconomy in the water sector through microalgae-based product recovery.

Performance of a green wall (Total Value Wall™) at high greywater loading rates and Life Cycle Impact Assessment.

Nature-based greywater (GW) treatment and reuse in urban areas has become an up-and-coming option. A 14.4 m2 green wall system called Total Value Wall (TVW) was installed at a terraced house in Gent (Belgium) for treating GW and reusing the effluent for toilet flushing. In a previous study, the TVW was loaded at 7 L.m-2.d-1 and efficiently removed TSS (67%), COD (43%), BOD5 (83%) and total coliforms (log 2), but a number of issues were reported related to nutrient leaching from the substrate, and the excessive retention time in the storage tanks. In this study results are reported from a follow-up study during which an adapted TVW was subjected to both higher hydraulic and pollutant loading rates in order to investigate the treatment capability of TVW. The design of the system, i.e. substrate contained in geotextile bags, did not sustain the higher hydraulic loading rates as excessive leakage occurred. Despite this, the higher pollutant loading rates still resulted in an acceptable effluent quality with 15 mg.L-1 TSS (90%), 85 mg.L-1 COD (82%), and 15 mg.L-1 BOD5 (95%). Ammonium, E. coli and total coliforms were removed with removal rates of 98%, 63% (0.4 log units), and 36% (0.2 log units), respectively. Finally, a life cycle assessment (LCA) was performed for the TVW with and without treating GW to analyze the environmental burden. The LCA impacts showed that replacing tap water and chemical fertilizer by GW, and the reuse of effluent, have a positive impact. However, the energy use for pumping has a major impact and should be minimized by using an efficient pump and distribution system to reduce the overall footprint.

Towards a general kinetic microalgae model: Extending a semi-deterministic green microalgae model for the cyanobacterium Arthrospira platensis and red alga Porphyridium purpureum.

Mathematical models for microalgae and cyanobacteria are seldomly validated for different algal species, as such limiting their applicability. Therefore, in this research, a previously developed kinetic model describing the growth of the green microalgae species Chlorella vulgaris was used to simulate the growth of the cyanobacterium Arthrospira platensis and the red alga Porphyridium purpureum. Based on a global sensitivity analysis, the model parameter µmax,A was calibrated using respirometric-titrimetric data. Calibration yielded values of 5.76 ± 0.17 d-1, 2.06 ± 0.16 d-1 and 1.06 ± 0.09 d-1 for Chlorella vulgaris, Arthrospira platensis and Porphyridium purpureum, respectively. Model simulations revealed that the biological growth equations in this model are adequate. However, increased light intensities triggered a survival mechanism for Arthrospira platensis, which is currently not taken into account by the model, leading to bad model accuracy under these circumstances. Future work should address the most important survival mechanisms and include those in the model to widen its applicability.

Model based analysis of carbon fluxes within microalgae-bacteria flocs using respirometric-titrimetric data.

With the emerging need of nutrient recycling in resource recovery facilities, the use of microalgae-bacteria flocs has received considerable attention in the past few years. However, although the main biological processes are already known, the complex interactions occurring between algae and bacteria are not fully understood. In this work, a combined respirometric-titrimetric unit was used to assess the microorganisms' kinetics within microalgae-bacteria flocs under different growth regimes (i.e. photoautotrophic, heterotrophic and mixotrophic) and different ratios of inorganic (IC) to organic carbon (OC) (IC:OC-ratios). Using this respirometric-titrimetric data, a new model was developed, calibrated and successfully validated. The model takes into account the heterotrophic growth of bacteria, the photoautotrophic, heterotrophic and mixotrophic growth of algae and the production and consumption of extracellular polymeric substances (EPS) by both bacteria and algae. As such, the model can be used for detailed analysis of the carbon fluxes within microalgae-bacteria flocs in an efficient way. Model analysis revealed the high importance of the EPS regulatory mechanism. Firstly, under heterotrophic growth conditions, OC-uptake occurred during the first 10-15 min. This was linked with internal OC storage (49% of added OC) and EPS production (40%), as such providing carbon reserves which can be consumed during famine conditions. Moreover, the algae were able to compete with bacteria for OC. Secondly, under photoautotrophic conditions, algae used the added IC to grow (57% of added IC) and to produce EPS (29%), which consecutively stimulated heterotrophic bacteria growth (20%). Finally, under mixotrophic conditions, low IC:OC-ratios resulted in an extensive OC-storage and EPS production (50% of added C) and an enhanced microalgal CO2 reuse, resulting in an increased algal growth of up to 29%.