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Toxic selenium oxyanions and sulfur species are often jointly present in contaminated waters and soils. This study investigated the effect on kinetics and resulting products for bio-reduction of selenium oxyanions in the presence of biologically produced sulfur resulting from bio-oxidation of sulfide in (bio)gas-desulfurization (bio-S0) and of sulfate. Selenite and selenate (~2 mmol L-1) bio-reduction was studied in batch up to 28 days at 30 oC and pH 7 using lactic acid and a sulfate-reducing sludge, 'Emmtec'. Bio-S0 addition increased the selenite removal rate, but initially slightly decreased selenate reduction rates. Selenite reacted with biologically generated sulfide resulting in selenium-sulfur, which upon further bio-reduction creates a sulfur bio-reduction cycle. Sulfate addition increased the bio-reduction rate for both selenite and sulfate. Bio-S0 or sulfate promoted hexagonal selenium formation, whereas without these, mostly amorphous Se0 resulted. With another inoculum, 'Eerbeek', bio-S0 accelerated the selenite reduction rate less than for 'Emmtec' because of lower sulfur and higher selenite bio-reduction rates. Bio-S0 addition increased the selenate reduction rate slightly and accelerated hexagonal selenium formation. Hexagonal selenium formation is advantageous because it facilitates separation and recovery and is less mobile and toxic than amorphous Se0. Insights into the interaction between selenium and sulfur bio-reduction are valuable for understanding environmental pathways and considerations regarding remediation and recovery.

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Recovery of selenium (Se) from wastewater provides a solution for both securing Se supply and preventing Se pollution. Here, we developed a high-rate process for biological selenate reduction to elemental selenium. Distinctive from other studies, we aimed for a process with selenate as the main biological electron sink, with minimal formation of methane or sulfide. A sequencing batch reactor, fed with an influent containing 120 mgSe L-1 selenate and ethanol as electron donor and carbon source, was operated for 495 days. The high rates (419 ± 17 mgSe L-1 day-1) were recorded between day 446 and day 495 for a hydraulic retention time of 6 h. The maximum conversion efficiency of selenate amounted to 96% with a volumetric conversion rate of 444 mgSe L-1 day-1, which is 6 times higher than the rates reported in the literature thus far. At the end of the experiment, a highly enriched selenate reducing biomass had developed, with a specific activity of 856 ± 26 mgSe-1day-1gbiomass-1, which was nearly 1000-fold higher than that of the inoculum. No evidence was found for the formation of methane, sulfide, or volatile reduced selenium compounds like dimethyl-selenide or H2Se, revealing a high selectivity. Ethanol was incompletely oxidized to acetate. The produced elemental selenium partially accumulated in the reactor as pure (≥80% Se of the total mixture of biomass sludge flocs and flaky aggregates, and ~100% of the specific flaky aggregates) selenium black hexagonal needles, with cluster sizes between 20 and 200 µm. The new process may serve as the basis for a high-rate technology to remove and recover pure selenium from wastewater or process streams with high selectivity.

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Bioelectrochemical systems (BES) can provide an energy efficient way to recover nutrients from wastewaters. However, the electron donors available in wastewater are often not sufficient to recover the total amount of nutrients. This work investigates hydrogen (H2) as an additional substrate for bioanodes. This hydrogen can be produced in the fermentation of complex organic waste or could be recycled from the cathode. Understanding how to influence the competition of electroactive microorganisms (EAM) with methanogens for H2 gas from different sources is key to successful application of H2 as additional electron donor in bioelectrochemical nutrient recovery. Ethanol (EtOH) was used as model compound for complex wastewaters since it is fermented into both acetate and H2. EtOH was efficiently converted into electricity (e-) by a syntrophic biofilm. Total recovered charge from 1 mM EtOH was 20% higher than for the same amount of acetate. This means that H2 from EtOH fermentation was converted by EAM into electricity. Low EtOH concentrations (1  mM) led to higher conversion efficiencies into electricity than higher concentrations (5 and 10  mM). Thermodynamic calculations show this correlates with a higher energy gain for electrogens compared to methanogens at low H2 concentrations. Cumulatively adding 1 mM EtOH without medium exchange (14 times in 14 days) resulted in stable conversion of H2 to e- (67%-77% e-) rather than methane. With H2 gas as electron donor, 68 ±â€¯2% H2 was converted into e- with no carbon source added, and still 53 ±â€¯5% to e- when 50  mM bicarbonate was provided. These results show that under the provided conditions, electrogens can outcompete methanogens for H2 as additional electron donor in MECs for nutrient recovery.

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In recent years, (bio)electrochemical systems (B)ES have emerged as an energy efficient alternative for the recovery of TAN (total ammonia nitrogen, including ammonia and ammonium) from wastewater. In these systems, TAN is removed or concentrated from the wastewater under the influence of an electrical current and transported to the cathode. Subsequently, it can be removed or recovered through stripping, chemisorption, or forward osmosis. A crucial parameter that determines the energy required to recover TAN is the load ratio: the ratio between TAN loading and applied current. For electrochemical TAN recovery, an energy input is required, while in bioelectrochemical recovery, electric energy can be recovered together with TAN. Bioelectrochemical recovery relies on the microbial oxidation of COD for the production of electrons, which drives TAN transport. Here, the state-of-the-art of (bio)electrochemical TAN recovery is described, the performance of (B)ES for TAN recovery is analyzed, the potential of different wastewaters for BES-based TAN recovery is evaluated, the microorganisms found on bioanodes that treat wastewater high in TAN are reported, and the toxic effect of the typical conditions in such systems (e.g., high pH, TAN, and salt concentrations) are described. For future application, toxicity effects for electrochemically active bacteria need better understanding, and the technologies need to be demonstrated on larger scale.

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Calcium phosphate (CaP) granules were discovered in the anaerobic treatment of vacuum collected black water (BW), using upflow anaerobic sludge blanket (UASB) technology. This allows simultaneous recovery of CaP granules and methane in the UASB reactor. However, the role of BW composition on CaP granulation is not yet understood. Moreover, CaP granulation was not observed in previous research on anaerobic treatment of BW, although similar treatment conditions were applied. Therefore, this study shows specifically the influence of bicarbonate and calcium fluctuations in BW on the phosphorus accumulation in the UASB reactor, which directly affects CaP granulation. Without calcium addition, 5% of the total phosphorus (P) fed was found as CaP granules in the reactor (61 mgP g-1dried matter), after 260 days of operation. Simultaneously, 65% of the COD in BW was efficiently converted into methane at 25 °C. Variations of bicarbonate and calcium concentrations in raw BW showed a significant influence on phosphorus accumulation in the UASB reactor. Geochemical modelling showed that the increase of soluble calcium from 39 to 54 mg L-1 in BW triggers supersaturation for calcium phosphate precursors (Cax(PO4)y). Concurrently, bicarbonate decreased from 2.7 to 1.2 g L-1, increasing further the ionic activity of calcium. Formation and accumulation of seed particles possibly enhanced CaP granulation. Preliminary results showed that addition of calcium (Ca2+/PO43- molar ratio of 3) increased the accumulation of total P in the UASB reactor to more than 85%. This further increases the granulation rate and consequently, the process feasibility.

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Microbial reduction of selenium sulfide (SeS2) is a key step in a new treatment process to recover selenium from selenate and selenite streams. In this process, selenate is first reduced to selenite, and subsequently selenite is reduced by sulfide and precipitates from the solution as SeS2. The latter is bio-reduced to elemental selenium and sulfide. Two anaerobic granular sludges (Eerbeek and Emmtec) were tested for their efficiency to reduce commercial crystalline SeS2. Emmtec sludge had the highest reducing capacity with commercial SeS2 and was therefore also used for the bioreduction of laboratory synthesized amorphous SeS2. Synthesized SeS2 was formed mixing a sulfide solution and effluent containing selenite. With both SeS2 solids (commercial and synthesized SeS2), Emmtec sludge produced sulfide and a solid consisting of hexagonal elemental selenium. The crystalline hexagonal structure suggests the absence of biomolecules, which stabilize amorphous selenium bio-particles under comparable process conditions (T=30°C and a pH between 6 and 7). Selenium particles were not attached to the biomass, suggesting an extracellular formation. The results support the feasibility of the bio-reduction process using sulfur for recovering selenium from water.

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An agar sediment test was developed to evaluate the suitability of organic waste streams from the food industry for recovering nutrients by the aquatic worm Lumbriculus variegatus (Lv). The effects of agar gel, sand, and food quantities in the sediment test on worm growth, reproduction, and water quality were studied. Agar gel addition ameliorated growth conditions by reducing food hydrolysis and altering sediment structure. Best results for combined reproduction and growth were obtained with 0.6% agar-gel (20 ml), 10 g. fine sand, 40 g. coarse sand, and 105 mg fish food (Tetramin). With agar gel, ingestion and growth is more the result of addition of food in its original quality. Final tests with secondary potato starch sludge and wheat bran demonstrated that this test is appropriate for the comparison of solid feedstuffs and suspended organic waste streams. This test method is expected to be suitable for organic waste studies using other sediment dwelling invertebrates.

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An innovative pilot-scale phototrophic biofilm reactor was evaluated over a 5-month period to determine its capacity to remove nitrogen and phosphorus from Dutch municipal wastewater effluents. The areal biomass production rate ranged between 2.7 and 4.5 g dry weight/m(2)/day. The areal nitrogen and phosphorus removal rates averaged 0.13 g N/m(2)/day and 0.023 g P/m(2)/day, which are low compared to removal rates achieved in laboratory biofilm reactors. Nutrient removal increased during the day, decreased with decreasing light intensity and no removal occurred during the night. Additional carbon dioxide supply was not requisite as the wastewater was comprised of enough inorganic carbon to sustain microalgal growth. The study was not conclusive for the limiting factor that caused the low nutrient removal rate, possibly the process was limited by light and temperature, in combination with pH increases above pH 9 during the daytime. This pilot-scale study demonstrated that the proposed phototrophic biofilm reactor is not a viable post-treatment of municipal wastewater effluents under Dutch climate conditions. However, the reactor performance may be improved when controlling the pH and the temperatures in the morning. With these adaptations, a phototrophic biofilm reactor could be feasible at lower latitudes with higher irradiance levels.

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Heptanoate (or enanthate), a saturated mono-carboxylate with seven carbon atoms, is a commercially produced biochemical building block with versatile applications. Currently, heptanoate is mainly derived from the oxidation of heptaldehyde, which can be obtained after pyrolysis of castor oil. The objective of this investigation was to achieve efficient high rate heptanoate production using a mixed culture chain elongation process based on propionate and ethanol. An efficient high rate heptanoate production using chain elongation could offer an alternative for heptanoate production from castor oil. The investigation was performed in an upflow anaerobic filter with a hydraulic retention time of 17 h. A heptanoate production rate of 4.5 g l(-1) d(-1) was achieved with a heptanoate concentration of 3.2 g l(-1). These results show sufficient potential to consider this approach as an alternative for heptanoate production from castor oil. Future research should make heptanoate production from propionate and ethanol more cost-effective.

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The objective of this investigation was to further increase the medium chain fatty acid (MCFA) production rate by reducing the hydraulic retention time (HRT) in an upflow anaerobic filter. The results showed that the volumetric MCFA production rate was increased to 57.4 g MCFA l(-1) d(-1), more than 3 times higher than previous work. Despite the lower MCFA concentrations at 4h HRT, the MCFA selectivity remained above 80%. Extra carbon dioxide additions and higher yeast extract concentrations were required to increase the MCFA production rate. More research related to substrates and (micro)nutrients in mixed culture continuous reactors needs to be performed to reduce yeast extract use in chain elongation.

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This research demonstrated the selective production of n-butyrate from mixed culture by applying 2 bar carbon dioxide into the headspace of batch fermenters or by increasing the initial substrate concentration. The effect of increasing initial substrate concentration was investigated at 8, 13.5 and 23 g COD/L with potato processing waste stream. Within 1 week of incubation, n-butyrate fraction selectively increased up to 83% by applying 2 bar hydrogen or 78% by applying carbon dioxide into the headspace whereas it was only 59% in the control reactor. Although the fraction of n-butyrate was elevated, the concentration remained lower than in the control. Both the highest concentration and fraction of n-butyrate were observed under the highest initial substrate concentration without headspace addition. The concentration was 10 g COD/L with 73% fraction. The operational conditions obtained from batch experiments for selective n-butyrate production were validated in a continuous process.

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Recently, interest has regained for medium chain fatty acids (MCFAs) as a low cost feedstock for bio-based chemical and fuel production processes. To become cost-effective, the volumetric MCFA production rate by chain elongation should increase to comparable rates of other fermentation processes. We investigate the MCFA production process at a hydraulic retention time of 17 h in an upflow anaerobic filter to improve the volumetric MCFA production rate. This approach resulted in a MCFA production with a volumetric production rate of 16.6 g l(-1) d(-1), which is more than seven times higher than the current production rate. Moreover the rate is now in the range of other fermentation processes like methane, butanol and ethanol production. Increasing the ethanol load lead to higher volumetric production rates and a high MCFA selectivity of 91%. During operation, methane percentages lower than 0.1% were detected in the headspace of reactor.

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This study investigated the carboxylate spectrum from mixed culture fermentation of three organic waste streams after supplying 2 bar hydrogen and carbon dioxide or a mixture of these two gases to the headspace. Under any modified headspace, propionate production was ceased and butyrate, caproate and the total carboxylate concentrations were higher than in the reactors with N(2) headspace (control). Production of one major compound was achieved under hydrogen and carbon dioxide mixed headspace after 4 weeks of incubation. Both the highest acetate concentration (17.4 g COD/l) and the highest fraction (87%) were observed in reactors with mixed hydrogen and carbon dioxide headspace independent of the substrate used. In the control reactor, acetate made up maximum 67% of the total products. For other products, the highest concentration and fraction were seldom observed together. Selective butyrate production reaching a 75% fraction was found under the carbon dioxide headspace on the carbohydrate rich waste.

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Nitrogen recovery through NH(3) stripping is energy intensive and requires large amounts of chemicals. Therefore, a microbial fuel cell was developed to simultaneously produce energy and recover ammonium. The applied microbial fuel cell used a gas diffusion cathode. The ammonium transport to the cathode occurred due to migration of ammonium and diffusion of ammonia. In the cathode chamber ionic ammonium was converted to volatile ammonia due to the high pH. Ammonia was recovered from the liquid-gas boundary via volatilization and subsequent absorption into an acid solution. An ammonium recovery rate of 3.29 g(N) d(-1) m(-2) (vs. membrane surface area) was achieved at a current density of 0.50 A m(-2) (vs. membrane surface area). The energy balance showed a surplus of energy 3.46 kJ g(N)(-1), which means more energy was produced than needed for the ammonium recovery. Hence, ammonium recovery and simultaneous energy production from urine was proven possible by this novel approach.

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In a Plant-Microbial Fuel Cell anode-conditions must be created that are favorable for plant growth and electricity production. One of the major aspects in this is the composition of the plant-growth medium. Hoagland medium has been used until now, with added phosphate buffer to reduce potential losses over the membrane because of differences in pH between anode and cathode. We developed a new, improved plant-growth medium that improves current production, while the plant keeps growing. This medium is a nitrate-less, ammonium-rich medium that contains all macro- and micro-nutrients necessary for plant growth, with a balanced amount of bicarbonate buffer. Sulphate presence in the plant-growth medium helps to keep a low anode-potential. With the new plant-growth medium the maximum current production of the Plant-Microbial Fuel Cell increased from 186 mA/m(2) to 469 mA/m(2).

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In order to meet environmental quality criteria, grey water was treated in four different ways: 1) aerobic 2) anaerobic+aerobic 3) aerobic+activated carbon 4) aerobic+ozone. Since each treatment has its own specific advantages and disadvantages, the aim of this study was to compare the ecotoxicity of differently treated grey water using Chironomus riparius (96 h test) and Daphnia magna (48 h and 21d test) as test organisms. Grey water exhibited acute toxicity to both test organisms. The aerobic and combined anaerobic+aerobic treatment eliminated mortality in the acute tests, but growth of C. riparius was still affected by these two effluents. Post-treatment by ozone and activated carbon completely removed the acute toxicity from grey water. In the chronic toxicity test the combined anaerobic+aerobic treatment strongly affected D. magna population growth rate (47%), while the aerobic treatment had a small (9%) but significant effect. Hence, aerobic treatment is the best option for biological treatment of grey water, removing most of the toxic effects of grey water. If advanced treatment is required, the treatment with either ozone or GAC were shown to be very effective in complete removal of toxicity from grey water.

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Source-separated black water (BW) (toilet water) containing 38% of the organic material and 68% of the phosphorus in the total household waste (water) stream including kitchen waste, is a potential source for energy and phosphorus recovery. The energy recovered, in the form of electricity and heat, is more than sufficient for anaerobic treatment, nitrogen removal and phosphorus recovery. The phosphorus balance of an upflow anaerobic sludge blanket reactor treating concentrated BW showed a phosphorus conservation of 61% in the anaerobic effluent. Precipitation of phosphate as struvite from this stream resulted in a recovery of 0.22 kgP/p/y, representing 10% of the artificial phosphorus fertiliser production in the world. The remaining part of the phosphorus ended up in the anaerobic sludge, mainly due to precipitation (39%). Low dilution and a high pH favour the accumulation of phosphorus in the anaerobic sludge and this sludge could be used as a phosphorus-enriched organic fertiliser, provided that it is safe regarding heavy metals, pathogens and micro-pollutants.

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Microalgal biofilms have so far received little attention as post-treatment for municipal wastewater treatment plants, with the result that the removal capacity of microalgal biofilms in post-treatment systems is unknown. This study investigates the capacity of microalgal biofilms as a post-treatment step for the effluent of municipal wastewater treatment plants. Microalgal biofilms were grown in flow cells with different nutrient loads under continuous lighting of 230 µmol/m(2)/s (PAR photons, 400-700 nm). It was found that the maximum uptake capacity of the microalgal biofilm was reached at loading rates of 1.0 g/m(2)/day nitrogen and 0.13 g/m(2)/day phosphorus. These maximum uptake capacities were the highest loads at which the target effluent values of 2.2 mg/L nitrogen and 0.15 mg/L phosphorus were still achieved. Microalgal biomass analysis revealed an increasing nitrogen and phosphorus content with increasing loading rates until the maximum uptake capacities. The internal nitrogen to phosphorus ratio decreased from 23:1 to 11:1 when increasing the loading rate. This combination of findings demonstrates that microalgal biofilms can be used for removing both nitrogen and phosphorus from municipal wastewater effluent.

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Ozonation and adsorption onto activated carbon were tested for the removal micropollutants of personal care products from aerobically treated grey water. MilliQ water spiked with micropollutants (100-1600 µgL(-1)) was ozonated at a dosing rate of 1.22. In 45 min, this effectively removed (>99%): Four parabens, bisphenol-A, hexylcinnamic aldehyde, 4-methylbenzylidene-camphor (4MBC), benzophenone-3 (BP3), triclosan, galaxolide and ethylhexyl methoxycinnamate. After 60 min, the removal efficiency of benzalkonium chloride was 98%, tonalide and nonylphenol 95%, octocrylene 92% and 2-phenyl-5-benzimidazolesulfonic acid (PBSA) 84%. Ozonation of aerobically treated grey water at an applied ozone dose of 15 mgL(-1), reduced the concentrations of octocrylene, nonylphenol, triclosan, galaxolide, tonalide and 4-methylbenzylidene-camphor to below limits of quantification, with removal efficiencies of at least 79%. Complete adsorption of all studied micropollutants onto powdered activated carbon (PAC) was observed in batch tests with milliQ water spiked with 100-1600 µgL(-1) at a PAC dose of 1.25 gL(-1) and a contact time of 5 min. Three granular activated carbon (GAC) column experiments were operated to treat aerobically treated grey water. The operation of a GAC column with aerobically treated grey water spiked with micropollutants in the range of 0.1-10 µgL(-1) at a flow of 0.5 bed volumes (BV)h(-1) showed micropollutant removal efficiencies higher than 72%. During the operation time of 1728 BV, no breakthrough of TOC or micropollutants was observed. Removal of micropollutants from aerobically treated grey water was tested in a GAC column at a flow of 2 BVh(-1). Bisphenol-A, triclosan, tonalide, BP3, galaxolide, nonylphenol and PBSA were effectively removed even after a stable TOC breakthrough of 65% had been reached. After spiking the aerobically treated effluent with micropollutants to concentrations of 10-100 µgL(-1), efficient removal to below limits of quantification continued for at least 1440 BV. Both ozonation and adsorption are suitable techniques for the removal of micropollutants from aerobically treated grey water.

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