Merle de Kreuk

Characterization of alginate-like exopolysaccharides isolated from aerobic granular sludge in pilot-plant.

To understand functional gel-forming exopolysaccharides in aerobic granular sludge, alginate-like exopolysaccharides were specifically extracted from aerobic granular sludge cultivated in a pilot plant treating municipal sewage. The exopolysaccharides were identified by the FAO/WHO alginate identification tests, characterized by biochemical assays, gelation with Ca(2+), blocks fractionation, spectroscopic analysis as UV-visible, FT-IR and MALDI-TOF MS, and electrophoresis. The yield of extractable alginate-like exopolysaccharides was reached 160+/-4mg/g (VSS ratio). They resembled seaweed alginate in UV-visible and MALDI-TOF MS spectra, and distinguished from it in the reactions with acid ferric sulfate, phenol-sulfuric acid and Coomassie brilliant blue G250. Characterized by their high percentage of poly guluronic acid blocks (69.07+/-8.95%), the isolated exopolysaccharides were capable to form rigid, non-deformable gels in CaCl(2). They were one of the dominant exopolysaccharides in aerobic granular sludge. We suggest that polymers play a significant role in providing aerobic granular sludge a highly hydrophobic, compact, strong and elastic structure.

Ecological characteristics of seeding sludge triggering a prompt start-up of anammox

Anammox start-up can be limited by the availability of seeding biomass in some areas. Previous studies have listed suitable alternative seeding sludge for anammox start-up such as anaerobic digestion sludge and conventional activated sludge (CAS), the ecological reasons behind has long been ignored. In this study, the inherent ecological factors that trigger a prompt start-up of anammox were identified, focusing on the initial relative abundance and concentration of anammox bacteria. An external membrane bioreactor was utilized as an enriching tool due to its suitability of retaining cells. Results revealed that a high initial concentration of anammox bacteria benefitted the start-up, meanwhile an even community seeding sludge (Gini coefficient<0.25) gained a more than three-time higher anammox activity compared to the uneven one (Gini coefficient>0.5). The discovery reminds to select the seeding sludge that is ecologically appropriate rather than to only care for the type of sludge in general.

Digester performance and microbial community changes in thermophilic and mesophilic sequencing batch reactors fed with the fine sieved fraction of municipal sewage

This study investigates the start-up and operation of bench-scale mesophilic (35 °C) and thermophilic (55 °C) anaerobic sequencing batch reactor (SBR) digesters treating the fine sieved fraction (FSF) from raw municipal sewage. FSF was sequestered from raw municipal wastewater, in the Netherlands, using a rotating belt filter equipped with a 350 micron mesh. For the given wastewater, the major component of FSF was toilet paper, which is estimated to be 10-14 kg per year per average person in the western European countries. A seven months adaptation time was allowed for the thermophilic and mesophilic digesters in order to adapt to FSF as the sole substrate with varying dry solids content of 10-25%. Different SBR cycle durations (14, 9 and 2 days) were applied for both temperature conditions to study methane production rates, volatile fatty acids (VFAs) dynamics, lag phases, as well as changes in microbial communities. The prevailing sludge in the two digesters consisted of very different bacterial and archaeal communities, with OP9 lineage and Methanothermobacter being pre-dominant in the thermophilic digester and Bacteroides and Methanosaeta dominating the mesophilic one. Eventually, decreasing the SBR cycle period, thus increasing the FSF load, resulted in improved digester performances, particularly with regard to the thermophilic digester, i.e. shortened lag phases following the batch feedings, and reduced VFA peaks. Over time, the thermophilic digester outperformed the mesophilic one with 15% increased volatile solids (VS) destruction, irrespective to lower species diversity found at high temperature.

Biomethanation from enzymatically hydrolyzed brewer's spent grain: Impact of rapid increase in loadings

Enzymatically hydrolyzed brewers spent grain (BSG) was digested in two expanded granular sludge beds (EGSBs, named BSG1 and BSG2, respectively). Both reactors were operated with the same organic loading rate (OLR) from 1 to 10kgCODm(-3)d(-1) during the first 45days. Hereafter a rapid OLR increase was applied to BSG2 from 10 to 16kgCODm(-3)d(-1) within three weeks, while the OLR of BSG1 was increased by less than 2kgCODm(-3)d(-1) in the same period. Results showed that a 30% decrease in COD removal and 70% decrease in methane yield appeared in BSG2 after the rapid OLR increase, and volatile fatty acid (VFA) accumulated more than thirty times compared to BSG1. The biomass structure deteriorated and 15% of the biomass was lost from the BSG2 reactor. 454-PyroTag and qPCR analysis revealed a rapid growth of acidifiers (i.e., Bacteroides) and a unique microbial community in BSG2 following the rapid increase in OLR.

Comparative analysis of the digestibility of sewage fine sieved fraction and hygiene paper produced from virgin fibers and recycled fibers

Sewage fine sieved fraction (FSF) is a heterogeneous substrate consisting of mainly toilet paper fibers sequestered from municipal raw sewage by a fine screen. In earlier studies, a maximum biodegradation of 62% and 57% of the sewage FSF was found under thermophilic (55°C) and mesophilic (35°C) conditions, respectively. In order to research this limited biodegradability of sewage FSF, this study investigates the biodegradation of different types of cellulosic fibers-based hygiene papers including virgin fibers based toilet paper (VTP), recycled fiber based toilet paper (RTP), virgin pulp for paper production (VPPP) as a raw material, as well as microcrystalline cellulose (MCC) as a kind of fiberless reference material. The anaerobic biodegradation or digestibility tests were conducted under thermophilic and mesophilic conditions. Results of the experiments showed different biomethane potential (BMP) values for each tested cellulose fiber-based substrate, which might be associated with the physical characteristics of the fibers, type of pulping, presence of lignin encrusted fibers, and/or the presence of additive chemicals and refractory compounds. Higher hydrolysis rates (Kh), higher specific methane production rates (SMPR) and shorter required incubation times to achieve 90% of the BMP (t90%CH4), were achieved under thermophilic conditions for all examined substrates compared to the mesophilic ones. Furthermore, the biodegradability of all employed cellulose fiber-based substrates was in the same range, 38-45%, under both conditions and less than the observed FSF biodegradability, i.e. 57-62%. MCC achieved the highest BMP and biodegradability, 86-91%, among all cellulosic substrates.

An integrated approach for efficient biomethane production from solid bio-wastes in a compact system.

BackgroundSolid bio-wastes (or organic residues) are worldwide produced in high amount and increasingly considered bioenergy containers rather than waste products. A complete bioprocess from recalcitrant solid wastes to methane (SW2M) via anaerobic digestion (AD) is believed to be a sustainable way to utilize solid bio-wastes. However, the complex and recalcitrance of these organic solids make the hydrolysis process inefficient and thus a rate-limiting step to many AD technologies. Effort has been made to enhance the hydrolysis efficiency, but a comprehensive assessment over a complete flow scheme of SW2M is rare.ResultsIn this study, it comes to reality of a complete scheme for SW2M. A novel process to efficiently convert organic residues into methane is proposed, which proved to be more favorable compared to conventional methods. Brewers’ spent grain (BSG) and pig manure (PM) were used to test the feasibility and efficiency. BSG and PM were enzymatically pre-hydrolyzed and solubilized, after which the hydrolysates were anaerobically digested using different bioreactor designs, including expanded granular sludge bed (EGSB), continuously stirred tank reactor (CSTR), and sequencing batch reactor (SBR). High organic loading rates (OLRs), reaching 19 and 21 kgCOD · m−3 · day−1 were achieved for the EGSBs, fed with BSG and PM, respectively, which were five to seven times higher than those obtained with direct digestion of the raw materials via CSTR or SBR. About 56% and 45% organic proportion of the BSG and PM can be eventually converted to methane.ConclusionsThis study proves that complex organic solids, such as cellulose, hemicellulose, proteins, and lipids can be efficiently hydrolyzed, yielding easy biodegradable/bio-convertible influents for the subsequent anaerobic digestion step. Although the economical advantage might not be clear, the current approach represents an efficient way for industrial-scale treatment of organic residues with a small footprint and fast conversion of AD.

Ethics in Innovation: Cooperation and Tension

An ethical parallel study has been carried out during the development of a new technology for wastewater treatment. Different aspects of the network involved in this innovation were studied. These include the inventory of the network, the risks in implementation of the technology foreseen by the different actors in the network and the responsibility for ethical issues, in particular the risks of the technology. Participating in a parallel study made the ethicists part of the network and influenced the discussions in the network, which indirectly affected the direction of the research. In this contribution the researchers reflect on the influence of the ethical parallel study on the innovation itself and on the proper role of ethicists in ethical parallel research.

Physical and biochemical changes in sludge upon Tubifex tubifex predation

ABSTRACT Worm predation (WP) on activated sludge leads to increased sludge degradation rates, irrespective of the type of worm used or reactor conditions employed. However, the cause of the increased sludge degradation rates remains unknown. This paper presents a comparative analysis of the physical and biochemical aspects of predated sludge, providing insight into the hydrolytic mechanisms underlying WP. To this end, the sessile worm Tubifex tubifex was used as a model oligochaete and was batch cultivated in an 18-L airlift reactor. Predation on activated sludge showed an average reduction rate of 12 ± 3.8%/d versus 2 ± 1.3%/d for endogenous respirated sludge. Sludge predation resulted in an increased release of inorganic nitrogen, phosphate and soluble chemical oxygen demand (sCOD). The sCOD consisted mainly of polysaccharides; however, fluorescence excitation emission matrix spectroscopy analysis also revealed the presence of Tryptophan-protein-like substances. Results suggest that the released polysaccharides contain a protein-like element. Additionally, soluble iron increased slightly in concentration after WP. The extent of hydrolysis seemed to reach an average plateau of about 40% volatile solids (VS) reduction after 4 days, which is substantially higher than the 29% VS reduction for endogenous decay of activated sludge after 30 days. Furthermore, T. tubifex predominantly consumed the protein fraction of the extracellular polymeric substances. Results suggest that that the worms specifically target a fraction of the sludge that is predominantly biodegradable under aerobic conditions, albeit at significantly higher degradation rates when compared to the endogenous decay of waste activated sludge.

Unravelling the protein preference of aquatic worms during waste activated sludge degradation

ABSTRACT Worm predation (WP) by Tubifex tubifex was investigated using waste activated sludge (WAS) as the substrate. In order to better understand the sludge degradation mechanisms during WP, the activity of five common hydrolytic enzymes was determined and compared among the initial feed activated sludge, endogenous respirated sludge and worm predated sludge. The results showed that the enzymatic activity decreased upon aerobic (worm) treatment of WAS and that this activity was predominantly associated with the removed solids fraction of the sludge. Interestingly, the protease activity showed a smaller decrease in activity when the worms were present. Flow cell cytometry revealed the release of intestinal bacteria from the worms, which are presumed to be largely responsible for the observed protease activity. Additionally, experiments in which T. tubifex were treated with antibiotics showed that the worms are responsible for a maximum of 73% of the observed proteolytic activity. The remaining 27% is attributed to the intestinal bacteria that exhibit a synergistic relationship with T. tubifex towards protein hydrolysis.

Impact of Coagulant and Flocculant Addition to an Anaerobic Dynamic Membrane Bioreactor (AnDMBR) Treating Waste-Activated Sludge

In this work, we investigated the effects of flocculation aid (FA) addition to an anaerobic dynamic membrane bioreactor (AnDMBR) (7 L, 35 °C) treating waste-activated sludge (WAS). The experiment consisted of three distinct periods. In period 1 (day 1–86), the reactor was operated as a conventional anaerobic digester with a solids retention time (SRT) and hydraulic retention time (HRT) of 24 days. In period 2 (day 86–303), the HRT was lowered to 18 days with the application of a dynamic membrane while the SRT was kept the same. In period 3 (day 303–386), a cationic FA in combination with FeCl3 was added. The additions led to a lower viscosity, which was expected to lead to an increased digestion performance. However, the FAs caused irreversible binding of the substrate, lowering the volatile solids destruction from 32% in period 2 to 24% in period 3. An accumulation of small particulates was observed in the sludge, lowering the average particle size by 50%. These particulates likely caused pore blocking in the cake layer, doubling the trans-membrane pressure. The methanogenic consortia were unaffected. Dosing coagulants and flocculants into an AnDMBR treating sludge leads to a decreased cake layer permeability and decreased sludge degradation.