Use of wood vinegar to enhance 5-aminolevulinic acid production by selected Rhodopseudomonas palustris in rubber sheet wastewater for agricultural use

This study aimed to produce inexpensive 5-aminolevulinic acid (ALA) in a non-sterile latex rubber sheet wastewater (RSW) by Rhodopseudomonas palustris TN114 and PP803 for the possibility to use in agricultural purposes by investigating the optimum conditions, and applying of wood vinegar (WV) as an economical source of levulinic acid to enhance ALA content. The Box–Behnken Design experiment was conducted under microaerobic-light conditions for 96 h with TN114, PP803 and their mixed culture (1:1) by varying initial pH, inoculum size (% v/v) and initial chemical oxygen demand (COD, mg/L). Results showed that the optimal condition (pH, % inoculum size, COD) of each set to produce extracellular ALA was found at 7.50, 6.00, 2000 for TN114; 7.50, 7.00, 3000 for PP803; and 7.50, 6.00, 4000 for a mixed culture; and each set achieved COD reduction as high as 63%, 71% and 75%, respectively. Addition of the optimal concentration of WV at mid log phase at 0.63% for TN114, and 1.25% for PP803 and the mixed culture significantly increased the ALA content by 3.7–4.2 times (128, 90 and 131 μM, respectively) compared to their controls. ALA production cost could be reduced approximately 31 times with WV on the basis of the amount of levulinic acid used. Effluent containing ALA for using in agriculture could be achieved by treating the RSW with the selected ALA producer R. palustris strains under the optimized condition with a little WV additive.     

The effect of methanogenesis inhibition,inoculum and substrate concentration on hydrogen and carboxylic acids production from cassava wastewater

Manipueira is a carbohydrate-rich agro-industrial waste from cassava processing. It is considered well suitable for biotechnological processes, such as hydrogen and carboxylic acids production, due to the high content of easily degradable organic matter. However, the proper methanogenesis inhibition method, inoculum type, and organic loads are factors still limiting the processes. The objective in this work was to evaluate the effects of such factors on byproducts production in anaerobic reactors. Batch experiments were conducted with 2.3-L flasks during two operational phases. In the first phase (P1), inhibition of methanogens in the sludge was evaluated using acetylene (1% v/v of headspace) and heat treatment (120 °C, 1 atm for 30 min). In the second phase (P2), three inoculum types obtained from common anaerobic sludges (bovine rumen and sludges from municipal and textile industrial wastewater treatment plants) were individually assayed. P2 aimed to identify the best inoculum, based on hydrogen production ability, which was tested for three initial concentrations of manipueira in terms of chemical oxygen demand (COD) (10, 20 and 40 g O₂/L). Results of P1 indicated that either acetylene or heat treatment efficiently inhibited methanogenesis, with no methane production. However, the maximum H₂ production potential by applying heat treatment (~ 563 mL) was more than twice compared with that by acetylene treatment (~ 257 mL); and butyrate was the main carboxylic acid by-product (~ 3 g/L). In P2 experiments after sludge heat treatment, the highest hydrogen yield (1.66 ± 0.07 mol H₂/mol glucose) and caproic acid production (~ 2 g/L) were observed at 20 g O₂/L of manipueira COD, when bovine rumen was the inoculum. The primary metabolic degradation products in all P2 experiments were ethanol, acetic, butyric, propionic and caproic acids. The finding of caproic acid detection indicated that the applied conditions in manipueira anaerobic degradation favored carbon chain elongation over methanogenesis.     

Bioremediation of synthetic high–chemical oxygen demand wastewater using microalgal species Chlorella pyrenoidosa

The microalgal species Chlorella pyrenoidosa was cultivated in synthetic wastewater of initial chemical oxygen demand (COD), nitrate, and phosphate concentrations of 5000, 100, and 40 mg/L, respectively. The aim of the study was to find out the tolerance of microalgae to different COD concentrations and the extent of COD degradation at those concentrations. Three dilutions of wastewater (initial COD concentrations 5000, 3000, and 1000 mg/L) and three inoculum sizes (0.1, 0.2, and 0.3 g/L) were considered for the study. The experimental parameters such as total organic carbon, total inorganic carbon, COD, optical density, total solids, nitrate, and phosphate were measured on a daily basis. Biodegradation kinetics was determined for all cases using first-order reaction and Monod degradation equations. Optimal results showed that up to 90% reduction in TOC was obtained for 1000 COD wastewater while only 38% reduction in total organic carbon (TOC) was achieved for 5000 COD wastewater. Over 95% reduction in nitrate and nearly 90% removal of phosphate were obtained with the lowest microalgal inoculum concentration (i.e., 0.1 g/L) for all COD dilutions. This study showed that microalgal species C. pyrenoidosa can successfully degrade the organic carbon source (i.e., acetate) with significant removal efficiencies for nitrate and phosphate.     

Statistical modeling and optimization of biomass granulation and COD removal in UASB reactors treating low strength wastewaters

The aim of this work was to study the influence of influent chemical oxygen demand (COD), upflow velocity of wastewater, and cationic polymer additives in inoculum, on biomass granulation and COD removal efficiency in upflow anaerobic sludge blanket (UASB) reactor for treating low strength wastewater. Statistical models were formulated based on these three variables to optimize the biomass granulation and COD removal efficiency in UASB reactors using a two-level, full factorial design. For the thick inoculum used in this study, having suspended solids (SS) >80 g/l and volatile suspended solids (VSS) to SS ratio <0.3, cationic polymer additives in the inoculum showed adverse effect on biomass granulation and COD removal efficiency. It is concluded that for such thick inoculum, granulation can be obtained while treating low strength wastewaters in UASB reactor by selecting proper combination of influent COD and liquid upflow velocity so as to represent the organic loading rate (OLR) greater than 1.0 kg COD/m(3) d. Validation of model predictions for treatment of synthetic wastewater and actual sewage reveals the efficacy of these models for enhancing granulation and COD removal efficiency.     

Effects of cationic polymer on performance of UASB reactors treating low strength wastewater

The effect of cationic polymer additives on biomass granulation and COD removal efficiency had been examined in lab-scale upflow anaerobic sludge blanket (UASB) reactors, treating low strength synthetic wastewater (COD 300-630 mg/l). Under identical conditions, two reactors were operated with and without polymer additives in inoculum under four different organic loading rates (OLRs). The optimum polymer dose was adopted based upon the results of jar test and settling test carried out with inoculum seed sludge. With the use of thick inoculum, SS greater than 110 g/l and VSS/SS ratio less than 0.3, granulation was observed in UASB reactor treating synthetic wastewater as well as actual sewage, when OLR was greater than 1.0 kg COD/m(3) d. Polymer additive with such thick inoculum was observed to deteriorate percentage granules and COD removal efficiency compared to inoculum without polymer additives. At OLR less than 1.0 kg COD/m(3) d, proper granulation could not be achieved in both the reactors inoculated with and without polymer additive. Also, under this low loading, drastic reduction in COD removal efficiency was observed with polymer additives in inoculum. Hence, it is rational to conclude that biomass granulation for treatment of low strength biodegradable wastewater depends on the applied loading rate and selection of thick inoculum sludge.     

Improvement of Biomethane Production from Sewage Sludge in Co-digestion with Glycerol and Waste Frying Oil,Using a Design of Experiments

A central composite design circumscribed method was used to define the experimental conditions that improve the methane production rate (k_CH4, liters of methane per kilogram of VS of waste added and per day) and the cumulative methane production (cMP, liters of methane per kilogram of VS of waste added) of the co-digestion of sewage sludge (SS) with crude glycerol (cGly) and waste frying oil (WFO). Three factors were selected, i.e., SS concentration, global co-substrate concentration, and mass fraction of cGly (x_cGly) in a mixture of cGly and WFO (in chemical oxygen demand, COD). SS digestion without co-substrate reached a cMP of (294?±?6) L·kg^?1 and a k_CH4 of (64?±?1) L·kg^?1·d^?1, at standard temperature and pressure conditions and expressed relatively to the initial volatile solids. After statistical analysis, SS and co-substrate concentrations of 4.6 g·L^?1 and 8.8 g·L^?1 (in COD), respectively, with x_cGly of 0.8, were defined to simultaneously boost cMP (91 % more) and k_CH4 (3-fold increase). Application of these conditions would yield 214 MWh more in electricity per 1000 m³ of SS digested.     

Kinetic Modelling and Characterization of Microbial Community Present in a Full-Scale UASB Reactor Treating Brewery Effluent

The performance of a full-scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was investigated by microbial analysis and kinetic modelling. The microbial community present in the granular sludge was detected using fluorescent in situ hybridization (FISH) and further confirmed using polymerase chain reaction. A group of 16S rRNA based fluorescent probes and primers targeting Archaea and Eubacteria were selected for microbial analysis. FISH results indicated the presence and dominance of a significant amount of Eubacteria and diverse group of methanogenic Archaea belonging to the order Methanococcales, Methanobacteriales, and Methanomicrobiales within in the UASB reactor. The influent brewery wastewater had a relatively high amount of volatile fatty acids chemical oxygen demand (COD), 2005 mg/l and the final COD concentration of the reactor was 457 mg/l. The biogas analysis showed 60–69 % of methane, confirming the presence and activities of methanogens within the reactor. Biokinetics of the degradable organic substrate present in the brewery wastewater was further explored using Stover and Kincannon kinetic model, with the aim of predicting the final effluent quality. The maximum utilization rate constant U _max and the saturation constant (K _B) in the model were estimated as 18.51 and 13.64 g/l/day, respectively. The model showed an excellent fit between the predicted and the observed effluent COD concentrations. Applicability of this model to predict the effluent quality of the UASB reactor treating brewery wastewater was evident from the regression analysis (R ²?=?0.957) which could be used for optimizing the reactor performance.     

Photosynthetic based algal-bacterial combined treatment of mixtures of organic pollutants and CO2 mitigation in a continuous photobioreactor

An algal-bacterial microcosm was synthetically constructed of Chlorella vulgaris MMl and Pseudomonas MTl. This microcosm was able to treat simulated wastewater supplemented with mixtures of phenol and pyridine up to 4.6 and 4.4 mM, respectively, in a continuous stirred tank bioreactor (CSTR) using photosynthetic oxygenation. Complete pollutant removal and detoxification and 82 % removal of introduced chemical oxygen demand (COD) were achieved at a hydraulic retention time (HRT) of 2.7 days. Increasing the influent load to 5.3 and 6.3 mM reduced the removal of phenol, pyridine and COD to 78, 21 and 59 %, respectively. Fertilization of the photobioreactor with 24 mM NaHCO₃ restored the treatment and detoxification efficiencies. The system was able to additionally mitigate up to 72 mM NaHCO₃ at the same HRT. Although the fertilization increased the system treatment efficiency, the settleability of the algal-bacterial microcosm was significantly reduced. When the photobioreactor was operated at HRT of 2.7 days in a 12/12 h of dark/light cycle, complete removal of 4.7 mM phenol was recorded but only 11 % of 5.7 mM pyridine was removed. The COD removal efficiency and CO₂ mitigation were also reduced to 65 and 86 %, respectively, and the effluent retained significant toxicity where 73 % inhibition was recorded. Elongation of the illumination time to 48 h (HRT of 4 days at 12/12 h dark/light cycle) restored the treatment and detoxification efficiencies.     

Cultivation of Microalgae in Dairy Farm Wastewater Without Sterilization

The present study investigated the feasibility of cultivating microalgae in dairy farm wastewater. The growth of microalgae and the removal rate of the nutrient from the wastewater were examined. The wastewater was diluted 20, 10 and 5 times before applied to cultivate microalgae. A 5 dilution yielded 0.86 g/L dry weight in 6 days with a relative growth rate of 0.28 d^?1, the 10× dilution gave 0.74 g/L and a relative growth rate of 0.26 d^?1 while the 20× dilution 0.59 g/L and a relative growth rate 0.23 d^?1. The nutrients in the wastewater could be removed effectively in different diluted dairy wastewater. The greatest dilution (20×) showed the removal rates: ammonia, 99.26%; P, 89.92%; COD, 84.18%. A 10× dilution removal% was: ammonia 93; P 91 and COD 88. The 5× dilution removal% was: ammonia 83; P 92; COD 90.     

SCP production and removal of organic load from cassava starch industry waste by yeasts

Wastewater obtained from the cassava processing industry was used to produce single cell protein (SCP). A few selected strains of yeasts were analysed for the SCP production based on the criteria of best biomass yield and high reduction in Chemical Oxygen Demand (COD) under nitrogen limitation. The monoculturing of Candida tropicalis gave the highest μmax with a protein enhancement of 4 fold and 50% decrease in COD. The performance of cocultures have shown an improvement in the COD removal of the wastewater and in the percent increase of protein. The effects of coculturing were better in a lab fermentor under controlled conditions.     

Fungal treatment followed by FeCl3 treatment to enhance enzymatic hydrolysis of poplar wood for high sugar yields

Fungal treatment followed by FeCl₃ treatment was used to improve saccharification of wood from Populus tomentosa. Combined treatments accumulated lignin and slightly degraded cellulose, whereas almost all hemicelluloses were removed. The white rot fungus, Trametes orientalis, and the brown rot fungus, Fomitopsis palustris, both accompanied by FeCl₃ post-treatment resulted in 98.8 and 99.7 % of hemicelluloses loss at 180 °C, respectively, which were over twice than that of hot water pretreatment at the same level. In addition, the solid residue from the T. orientalis-assisted and F. palustris-assisted FeCl₃ treatment at 180 °C released 84.5 and 95.4 % of reducing sugars, respectively: 1.4- and 1.6-fold higher than that of FeCl₃ treatment alone at the same temperature. Combined treatments disrupted the intact cell structure and increased accessible surface area of cellulose therefore enhancing the enzymatic digestibility, as evidenced by XRD and SEM analysis data.     

Anti-inflammatory activity of phlorotannin-rich fermented Ecklonia cava processing by-product extract in lipopolysaccharide-stimulated RAW 264.7 macrophages

In this study, fermentation was employed as a tool to further increase the bioactive potential of processing by-product from a brown seaweed, Ecklonia cava, which can be obtained from food and cosmetic industries after its polyphenol extraction. The fermentation process was done for 24 h using an industrially important microorganism Candida utilis prior to being extracted with 80 % ethanol. The anti-inflammatory potential of the fermented E. cava processing by-product extract (FEPBE) was evaluated in vitro. The phlorotannin-rich FEPBE dose-dependently inhibited the nitric oxide production, prostaglandin-E₂ production and suppressed the inducible nitric oxide synthase and cyclooxygenase-2 expressions in lipopolysaccharide-stimulated RAW 264.7 cells. The release of pro-inflammatory cytokines, interleukin-1β and interleukin-6, was significantly suppressed by the extract in a dose-dependent manner. Due to the profound anti-inflammatory activity, FEPBE appears as a value-added biomass fraction that can be exploited in numerous industrial applications as a source of functional ingredients.     

Characterization and application of bioflocculant prepared by Rhodococcus erythropolis using sludge and livestock wastewater as cheap culture media

A new bioflocculant was produced by culturing Rhodococcus erythropolis in a cheap medium. When culture pH was 7.0, inoculum size was 2 % (v/v), Na₂HPO₄ concentration was 0.5 g L^?1, and the ratio of sludge/livestock wastewater was 7:1 (v/v), a maximum flocculating rate of 87.6 % could be achieved. Among 13 different kinds of pretreatments for sludge, the optimal one was the thermal-alkaline pretreatment. Different from a bioflocculant produced in a standard medium, this bioflocculant was effective over a wide pH range from 2 to 12 with flocculating rates higher than 98 %. Approximately, 1.6 g L^?1 of crude bioflocculant could be harvested using cold ethanol for extraction. This bioflocculant showed color removal rates up to 80 % when applied to direct and disperse dye solutions, but only 23.0 % for reactive dye solutions. Infrared spectrum showed that the bioflocculant contained functional groups such as –OH, –NH₂, and –CONH₂. Components in the bioflocculant consisted of 91.2 % of polysaccharides, 7.6 % of proteins, and 1.2 % of DNA. When the bioflocculant and copper sulfate (CuSO₄) were used together for decolorization in actual dye wastewater, the optimum decolorization conditions were specified by the response surface methodology as pH 11, bioflocculant dosage of 40 mg/L, and CuSO₄ 80 mg/L, under which a decolorization rate of 93.9 % could be reached.     

The effects of light intensity, inoculum size, and cell immobilisation on the treatment of sago effluent withRhodopseudomonas palustris strain B1

A study was carried out to determine a suitable light intensity and inoculum size for the growth ofRhodopseudomonas palustris strain B1. The pollution reduction of sago effluent using free and immobilisedR. palustris cells was also evaluated. The growth rate in glutamatemalate medium was highest at 4 klux compared to 2.5 and 3 klux. The optimal inoculum size was 10% (v/v). Both the COD and BOD of the sago effluent were reduced by 67% after three days of treatment. The difference in biomass production or BOD and COD removal with higher inoculum sizes of 15 and 20% was minimal. This could be attributed to limited nutrient availabillity in the substrate. The use of immobilised cells ofR. palustris reduced the pollution load 10% less compared to pollution reduction by free cells. Hence, there was no significant difference in using free or immobilised cells for the treatment of sago effluent.     

Biological treatment of saline wastewater in a rotating biodisc contactor by using halophilic organisms

Biological treatment of saline wastewater presents unique difficulties as a result of plasmolysis of microorganisms in the presence of salt. Removal of salt from wastewater before biological treatment by reverse osmosis or ion exchange operations are rather expensive. Inclusion of halophilic organisms in activated sludge culture seems to be a more practical approach in biological treatment of saline wastewater. A synthetic wastewater composed of diluted molasses, urea, KH₂PO₄, MgSO₄ and various concentrations of salt (0–5% NaCl) was treated in a rotating biodisc contactor (RBC). A salt tolerant organism Halobacter halobium was added onto activated sludge culture (50%) and used as inoculum. Effects of important process variables such as A/Q ratio, COD loading rate, feed COD concentration, salt concentration and liquid phase aeration on system performance were investigated. An empirical mathematical model describing the system's performance as a function of important process variables was developed and constants were determined by using the experimental data.     

Electricity generation from young landfill leachate in a microbial fuel cell with a new electrode material

This study aims at evaluating the performance of a two-chambered continuously fed microbial fuel cell with new Ti–TiO₂ electrodes for bioelectricity generation from young landfill leachate at varying strength of wastewater (1–50 COD g/L) and hydraulic retention time (HRT, 0.25–2 days). The COD removal efficiency in the MFC increased with time and reached 45 % at full-strength leachate (50 g/L COD) feeding. The current generation increased with increasing leachate strength and decreasing HRT up to organic loading rate of 100 g COD/L/day. The maximum current density throughout the study was 11 A/m² at HRT of 0.5 day and organic loading rate of 67 g COD/L/day. Coulombic efficiency (CE) decreased from 57 % at feed COD concentration of 1 g/L to less than 1 % when feed COD concentration was 50 g/L. Increase in OLR resulted in increase in power output but decrease in CE.     

Production of hydrogen from domestic wastewater in a pilot-scale microbial electrolysis cell

Addressing the need to recover energy from the treatment of domestic wastewater, a 120-L microbial electrolysis cell was operated on site in Northern England, using raw domestic wastewater to produce virtually pure hydrogen gas (100?±?6.4 %) for a period of over 3 months. The volumetric loading rate was 0.14 kg of chemical oxygen demand (COD) per cubic metre per day, just below the typical loading rates for activated sludge of 0.2–2 kg?COD?m^?3?day^?1, at an energetic cost of 2.3 kJ/g?COD, which is below the values for activated sludge 2.5–7.2 kJ/g?COD. The reactor produced an equivalent of 0.015 L?H₂?L^?1?day^?1, and recovered around 70 % of the electrical energy input with a coulombic efficiency of 55 %. Although the reactor did not reach the breakeven point of 100 % electrical energy recovery and COD removal was limited, improved hydrogen capture and reactor design could increase the performance levels substantially. Importantly, for the first time, a ‘proof of concept’ has been made, showing that this technology is capable of energy capture as hydrogen gas from low strength domestic wastewaters at ambient temperatures.     

Biotreatment of heavy oil wastewater by combined upflow anaerobic sludge blanket and immobilized biological aerated filter in a pilot-scale test

A field pilot study has been constructed in the Liaohe oilfield, China to treat heavy oil wastewater with large amounts of dissolved recalcitrant organic compounds and low nutrient of nitrogen and phosphorus by an upflow anaerobic sludge blanket (UASB) coupled with immobilized biological aerated filters (I-BAFs). By operating the system for 252 days (including the start-up of 128 days), the chemical oxygen demand (COD), ammonia nitrogen (NH₃-N) and suspended solid (SS) in the wastewater were removed by 74%, 94% and 98%, respectively. GC–MS analysis indicated that most of alkanes were degraded by the UASB process, while the I-BAF played important roles both in degrading organic compounds and in removing the NH₃-N and SS. The bacterial community structural analysis based on the PCR-DGGE technology reveals that the predominant bacteria in the UASB reactor belong to the Bacillales and Rhodobacterales, and that in the I-BAF was identified as uncultured soil bacterium. Our results suggest that the combined biotreatment system has immense potential in large-scale treatment of heavy oil wastewater.     

Isolation and selection of native microorganisms for the aerobic treatment of simulated dairy wastewaters

Milk fat/protein degrading microorganisms were isolated from different locations of a dairy wastewater treatment system with the goal of developing an inoculum for bioaugmentation strategies. Eight isolates, identified by 16S rRNA gene sequence analysis as belonging to the genera Bacillus, Pseudomonas, and Acinetobacter, were tested for their ability to remove COD and protein from a milk-based medium (3000 mg/L COD) and compared to a commercial bioaugmentation inoculum. The Acinetobacter isolate exhibited a pellet-type growth in liquid culture, a property that could potentially aid in the separation of microbes and liquid phase following treatment. Based on the individual degradation capacity and growth behavior of the isolates, three microorganisms were further selected and tested together. This consortium exhibited a COD removal similar to the commercial inoculum (57% and 63%, respectively), but higher protein (consortium: 93%; commercial inoculum: 54%), and fat removals (consortium: 75%; commercial inoculum: 38%).     

Utilization of orange peel,a food industrial waste,in the production of exo-polygalacturonase by pellet forming <Emphasis Type="Italic">Aspergillus sojae</Emphasis>

The production of exo-polygalacturonase (exo-PG) from orange peel (OP), a food industrial waste, using Aspergillus sojae was studied in submerged culture. A simple, low-cost, industrially significant medium formulation, composed of only OP and (NH₄)₂SO₄ (AS) was developed. At an inoculum size of 2.8 × 10³ spores/mL, growth was in the form of pellets, which provided better mixing of the culture broth and higher exo-PG activity. These pellets were successfully used as an inoculum for bioreactors and 173.0 U/mL exo-PG was produced. Fed-batch cultivation further enhanced the exo-PG activity to 244.0 U/mL in 127.5 h. The final morphology in the form of pellets is significant to industrial fermentation easing the subsequent downstream processing. Furthermore, the low pH trend obtained during this fermentation serves an advantage to fungal fermentations prone to contamination problems. As a result, an economical exo-PG production process was defined utilizing a food industrial by-product and producing high amount of enzyme.