Performance of an ECSB reactor for high-rate anaerobic treatment of cheese industry wastewater: effect of pre-acidification on process efficiency and calcium precipitation.

An external circulation sludge bed (ECSB) reactor was studied at full-scale (1,000 m3) during anaerobic treatment of cheese and other dairy products industry wastewater (CWW). The latter was characterized by a high calcium content, therefore the study focused on the potential negative impact that calcium may have in the long-term. The degree of CWW acidification (25 and 40%) on ECSB reactor performance was evaluated over a wide range of organic loading rates from 5 up to 18 kg m3 d-1, while process efficiency and calcium precipitation were examined in detail. Independently of the operating conditions, the volatile suspended solids content of the anaerobic granular sludge, as well as its calcium content, remained stable along the ECSB reactor operation, indicating that there was no calcium build up in the biomass. The results of this study demonstrate that the ECSB design seems to be particularly suitable to treat calcium-rich wastewater that is probably due to the fact that in this system CaCO3 precipitates in the bulk liquid of the external circulation tank and not the biomass present in the main reactor, and that the CaCO3 crystals are washed-out from it due to the high upflow velocity applied to the system (5 m h-1).

Inhibition of the respiratory chain reactions in denitrifying EBPR biomass under simultaneous presence of acetate and electron acceptor.

In this study, the deterioration of the typical EBPR (Enhanced Biological Phosphorus Removal) process due to the simultaneous presence of electron donor (external substrate) and electron acceptor (oxygen or nitrate) was investigated by using a PAOs (Polyphosphate Accumulating Organisms)-enriched biomass grown in a modified DEPHANOX system. Intracellular and extracellular constituents were monitored in batch tests under different electron donor and acceptor conditions and specific oxygen and nitrogen uptake rates were evaluated. Results showed that phosphorus uptake was inhibited during the simultaneous presence of electron donor (acetate) and acceptor (O2/NO3-) in the mixed liquor. In the presence of acetate, PHAs and glycogen were produced under both aerobic and anoxic conditions irrespectively to the PHAs amount already stored intracellularly. The Krebs cycle reactions and oxidative phosphorylation provided the reduced coenzymes and energy required for PHAs synthesis when biomass polyphosphate content was low. On the contrary, polyphosphate cleavage provided the ATP required for PHAs synthesis in the presence of high biomass polyphosphate content. Inhibition of the respiratory chain reactions was observed when biomass with high polyphosphate and low PHAs content was subjected to simultaneous presence of electron donor and acceptor. PHAs utilization rather than glycogen degradation appears to favor phosphate accumulation since no polyphosphate synthesis occurred in the absence of PHAs reserves.

Performance and metabolic aspects of a novel enhanced biological phosphorus removal system with intermittent feeding and alternate aeration.

A novel enhanced biological phosphorus removal (EBPR) system, which combined the intermittent feeding design with an anaerobic selector, was examined using on-line oxidation reduction potential (ORP), nitrate and ammonium probes. Two experimental periods were investigated: the aerobic and anoxic phases were set at 40 and 20 minutes respectively for period I, and set at 30 and 30 minutes for period II. Chemical oxygen demand (COD), biochemical oxygen demand (BOD5) and P removal were measured as high as 87%, 96% and 93% respectively, while total Kjeldahl nitrogen (TKN) and NH4(+) removal averaged 85% and 91%. Two specific denitrification rates (SDNRs), which corresponded to the consumption of the readily biodegradable and slowly biodegradable COD, were determined. SDNR-1 and SDNR-2 during period I were 0.235 and 0.059 g N g(-1) volatile suspended solids (VSS) d(-1) respectively, while the respective rates during period II were 0.105 and 0.042 g N g(-1) VSS d(-1). The specific nitrate formation and ammonium oxidizing rates were 0.076 and 0.064 g N g(-1) VSS d(-1) for period I and 0.065 and 0.081 g N g(-1) VSS d(-1) for period II respectively. The specific P release rates were 2.79 and 4.02 mg P g(-1) VSS h(-1) during period I and II, while the respective anoxic/aerobic uptake rates were 0.42 and 0.55 mg P g(-1) VSS h(-1). This is the first report on an EBPR scheme using the intermittent feeding strategy.

Metabolic behavior and enzymatic aspects of denitrifying EBPR sludge in a continuous-flow anaerobic-anoxic system.

The metabolic aspects of enhanced biological phosphorus removal (EBPR) were investigated for the first time in a continuous-flow anaerobic-anoxic plant fed with acetate, propionate, or substrates which are involved in the tricarboxylic acid and/or glyoxylate cycle, i.e., fumarate, malate, or oxaloacetate, as the sole carbon source. Although the polyphosphate-accumulating organisms (PAOs) population remained stable with any carbon source examined, no typical EBPR metabolism was observed during fumarate, malate, or oxaloacetate utilization. Specific enzymatic activities related to EBPR were determined in activated sludge homogenates and directly correlated with the nutrient metabolic rates. The experimental results indicated the direct involvement of alkaline phosphatase, pyrophosphatase, and exopolyphosphatase in the denitrifying EBPR process. Metabolic aspects of glyoxylate cycle enzymes are discussed with regard to the biomass anaerobic and anoxic activity. Process performance was highly influenced by the kind of substrate utilized, indicating that specific metabolic pathways should be followed to favor efficient EBPR.

Intermittent operation of low pressure UF membranes for sewage reuse at household level.

A household-scale wastewater treatment system was operated with domestic sewage. The system could recover gardening/irrigation water from raw sewage or secondary effluent by low pressure ultrafiltration (UF). The UF membranes (surface area = 3.5 m(2), pore size = 0.04 µm) were operated at constant transmembrane pressure (0.13 bar). The proposed technology was examined for approximately 2 months without membrane cleaning. Membrane operation was performed periodically (one or two times per week), simulating water usage for gardening irrigation. During raw sewage filtration (chemical oxygen demand (COD) total = 242 ± 71 mg L(-1), COD soluble = 105 ± 51 mg L(-1), suspended solids = 188 ± 58 mg L(-1)), low permeate COD was achieved (52 ± 25 mg L(-1)), whereas nitrogen and phosphorus were recovered in the permeate. The water recovered during 1 h of operation displayed a gradual decrease from 42 to 22 L m(-2)h(-1) during the 50-d time period. For the secondary effluent filtration, the UF module achieved consistently a recovery rate of 39.6 ± 8.0 L m(-2)h(-1), with an average permeate COD of 37 mg L(-1). In this case, the fouling layer (cake layer) was completely reversible after the relaxation period, rendering the process suitable for unattended household applications.

Continuous biohydrogen production from fruit wastewater at low pH conditions.

Biohydrogen production from a simulated fruit wastewater (soluble COD = 3.17 ± 0.10 g L⁻¹) was carried out in a continuous stirred tank reactor (CSTR) of 2 L operational volume without biomass inoculation, heat pre-treatment or pH adjustment, resulting in a low operational pH (3.75 ± 0.09). The hydraulic retention time (HRT) varied from 15 to 5 h. A strong negative correlation (p < 0.01) between the biogas production rate and the HRT was observed. Biogas production rates were higher at 30 °C than at 25 °C (p < 0.01), when the CSTR was operated under the same HRT. The biogas hydrogen content was estimated as high as 55.8 ± 2.3 % and 55.4 ± 2.5 % at 25 and 30 °C, respectively. The main fermentation end products were acetic and butyric acids, followed by ethanol. Significant differences (p < 0.01) during the operation of the CSTR at 25 or 30 °C were identified for butyric acid at almost all HRTs examined. Simulation of the acidogenesis process in the CSTR (based on COD and carbon balances) indicated the possible metabolic compounds produced at 25 and 30 °C reactions and provided an adequate fit of the experimental data.

Molecular characterization of denitrifying bacteria isolated from the anoxic reactor of a modified DEPHANOX plant performing enhanced biological phosphorus removal.

Enhanced Biological Phosphorus Removal (EBPR) under anoxic conditions was achieved using a Biological Nutrient Removal (BNR) system based on a modification of the DEPHANOX configuration. Double-probe Fluorescence in Situ Hybridization (FISH) revealed that Polyphosphate Accumulating Organisms (PAOs) comprised 12.3 +/- 3.2% of the total bacterial population in the modified DEPHANOX plant. The growing bacterial population on blood agar and Casitone Glycerol Yeast Autolysate agar (CGYA) medium was 16.7 +/- 0.9 x 10(5) and 3.0 +/- 0.6 x 10(5) colony forming units (cfu) mL(-1) activated sludge, respectively. A total of 121 bacterial isolates were characterized according to their denitrification ability, with 26 bacterial strains being capable of reducing nitrate to gas. All denitrifying isolates were placed within the alpha-, beta-, and gamma-subdivisions of Proteobacteria and the family Flavobacteriaceae. Furthermore, a novel denitrifying bacterium within the genus Pseudomonas was identified. This is the first report on the isolation and molecular characterization of denitrifying bacteria from EBPR sludge using a DEPHANOX-type plant.

Denitrifying polyphosphate accumulating organisms population and nitrite reductase gene diversity shift in a DEPHANOX-type activated sludge system fed with municipal wastewater.

Enhanced biological phosphorus removal (EBPR) is a widely applied method for nutrients removal, although little is known about the key genes regulating the complex biochemical transformations occurring in activated sludge during phosphorus removal. In the present study, the nitrite reductase gene (nirS) diversity and the denitrifying polyphosphate accumulating organisms (DPAOs) population, grown in a bench scale, two-sludge, continuous flow plant, operating for biological anoxic phosphorus removal (DEPHANOX-type), fed with municipal wastewater, were examined by means of physicochemical analyses and the application of molecular techniques. The DEPHANOX configuration highly influenced biomass phosphorus as well as polyhydroxyalkanoates content and facilitated the enrichment of the DPAOs population. The application of double probe fluorescent in situ hybridization (double probe FISH) technique revealed that DPAOs comprised 20% of the total bacterial population. Based on clone libraries construction and nirS gene sequencing analysis, a pronounced shift in denitrifying bacteria diversity was identified during activated sludge acclimatization. Moreover, nirS gene sequences distinct from those detected in any known bacterial strain or environmental clone were identified. This is the first report studying the microbial properties of activated sludge in a DEPHANOX-type system using molecular techniques.

Intermittent feeding of wastewater in combination with alternating aeration for complete denitrification and control of filaments.

In the present study, a laboratory scale system, consisting of a primary settling tank, a continuous stirred tank reactor and a clarifier were constructed and operated, using wastewater from the municipal wastewater treatment plant in Xanthi, Greece. The system operated under intermittent aeration in aerobic/anoxic conditions and feeding of the wastewater once in every cycle. The unit was inoculated with sludge, which originated from the recirculation stream of the local wastewater treatment plant. The wastewater was processed with hydraulic retention time (HRT) of 12 h, in which various experimental states were studied regarding the combination of aerobic and anoxic intervals. The wastewater was fed in limited time once in every cycle of aerobic/anoxic conditions at the beginning of the anoxic period. The two states that exhibited highest performance in nitrification and total nitrogen removal were, then, repeated with HRT of 10 h. The results show that, regarding the nitrification stage and the organic load removal, the intermittent system achieved optimum efficiency, with an overall removal of biological oxygen demand (BOD(5)) and ammonium nitrogen in the range of 93-96% and 91-95% respectively. As far as the total nitrogen removal is concerned, and if the stage of the denitrification is taken into account, the performance of the intermittent system surpassed other methods, as it is shown by the total Kjeldahl nitrogen (TKN) removal efficiency of 85-87%. These operating conditions suppressed the growth of filamentous organisms, a fact reflected at the SVI values, which were lower than 150 ml/g.

High aeration rate enhances flow stratification in full-scale oxidation ditch.

Aerated channel reactors with a uniform field of aeration may display flow stratification and short-circuit phenomena in wastewater treatment systems. In this study, we present data suggesting that flow stratification is closely related to the aeration rate and the arrangement of aerators. A full-scale oxidation ditch, with a total volume of 6,500 m(3) and a membrane-diffused aerated zone of 60 x 7 x 5 m (length-width-depth), was selected for water velocity measurements. Two profiles of the oxidation ditch were studied in detail: the first one was at the end of the aerated zone and the second one at the end of the anoxic zone. The results of this work demonstrate that the horizontal water velocity at the end of the aerated zone displayed significant stratification, with maximum velocity near the water surface (0.5-0.7 m/s) and almost zero velocity at a depth of 2.5 m. At the end of the anoxic zone, water velocity was uniform and equal to 0.27-0.31 m/s. Increasing the aeration rate from 1,800 to 4,300 m(3)/h, almost 90% of the water flow was found to discharge through the upper-half of the channel reactor profile. Different options to mitigate flow stratification of the oxidation ditch are discussed in this paper.

Sulfide removal in wastewater from petrochemical industries by autotrophic denitrification.

An alternative flowchart for the biological removal of hydrogen sulfide from oil-refining wastewater is presented; autotrophic denitrification in a multi-stage treatment plant was utilized. A pilot-scale plant was fed with a mixture of the following constituents: (a) original wastewater from an oil refining industry (b), the effluent of the existing nitrification-stage treatment plant and (c) sulfide in the form of Na2S. Anoxic sulfide to sulfate oxidation, with nitrate as a terminal electron acceptor, proved very successful, as incoming concentrations of 110 mg S2-/L were totally converted to SO(4)2-. At complete denitrification, the concentration of S2- in the reactor effluent was less than 0.1mg/L. Fluctuating S2- concentration in the feed could be tolerated without any problems, as the accumulated sulfide in the effluent of the denitrification stage is oxidized aerobically in a subsequent activated-sludge treatment stage. This alternative new treatment scheme was further introduced at the refinery's wastewater processing plant. Thus, complete H2S removal is now accomplished by the combination of the proposed biological method and the existing stripping with CO2. As a result, stripping, and thus its cost, is reduced by 70%.

Process optimization of continuous gluconic acid fermentation by isolated yeast-like strains of Aureobasidium pullulans.

This study was focused on the optimization of a new fermentation process for continuous gluconic acid production by the isolated yeast-like strain Aureobasidium pullulans DSM 7085 (isolate 70). Operational fermentation parameters were optimized in chemostat cultures, using a defined glucose medium. Different optima were found for growth and gluconic acid production for each set of operation parameters. Highest productivity was recorded at pH values between 6.5 and 7.0 and temperatures between 29 and 31 degrees C. A gluconic acid concentration higher than 230 g/L was continuously produced at residence times of 12 h. A steady state extracellular gluconic acid concentration of 234 g/L was measured at pH 6.5. 122% air saturation yielded the highest volumetric productivity and product concentration. The biomass-specific productivity increased steadily upon raising air saturation. An intracellular gluconic acid concentration of about 159 g/L (0.83 mol) was determined at 31 degrees C. This is to be compared with an extracellular concentration of 223 g/L (1.16 mol), which indicates the possible existence of an active transport system for gluconic acid secretion, or the presence of extracellular glucose oxidizing enzymes. The new process provides significant advantages over the traditional discontinuous fungi operations. The process control becomes easier, thus offering stable product quality and quantity.

Biochemical reaction engineering and process development in anaerobic wastewater treatment.

Developments in production technology have frequently resulted in the concentrated local accumulation of highly organic-laden wastewaters. Anaerobic wastewater treatment, in industrial applications, constitutes an advanced method of synthesis by which inexpensive substrates are converted into valuable disproportionate products. A critical discussion of certain fundamental principles of biochemical reaction engineering relevant to the anaerobic mode of operation is made here, with special emphasis on the roles of thermodynamics, kinetics, mass and heat transfer, reactor design, biomass retention and recycling. The applications of the anaerobic processes are discussed, introducing the principles of an upflow anaerobic sludge bed reactor and a fixed-bed loop reactor. The merits of staging reactor systems are presented using selected examples based on two decades of research in the field of anaerobic fermentation and wastewater treatment at the Forschungszentrum Julich (Julich Research Center, Germany). Wastewater treatment is an industrial process associated with one of the largest levels of mass throughput known, and for this reason it provides a major impetus to further developments in bioprocess technology in general.