Relationship of protozoan biomass to phosphate and nitrate removal from activated sludge mixed liquor

The relationship between protozoan biomass concentration and phosphate and nitrate removal was investigated in mixed liquor using three different carbon sources as supplements. The study was carried out using three respective initial biomass concentrations in a shaking flask environment. Samples were taken every 24 h to determine phosphate, nitrate, dissolved oxygen and chemical oxygen demand. The results revealed a direct relationship between decreases in nutrient concentrations and increases in cell densities of the isolates. Between 24 and 96 h, the increases in the protozoan density corresponded to a phosphate decreases from initial ranges of 55.42–57.36 mg/L, 50.27–51.17 mg/L and 50.01–50.83 mg/L to final ranges of 2.46–11.90 mg/L, 0.61–11.80 mg/L and 1.29–13.89 mg/L, in the presence of Aspidisca, Trachelophyllum and Peranema, respectively. Nitrate concentrations were observed to decrease from initial ranges of 23.84–25.90 mg/L, 23.94–25.84 mg/L and 26.12–26.54 mg/L to final ranges of 0.11–6.32 mg/L, 0.16–5.60 mg/L and 0.24–9.04 mg/L, respectively. The study had revealed that an increase in cell density of the test isolates produces a corresponding increase in phosphate and nitrate removal.     

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.     

Biomass production from glutamate fermentation wastewater by the co-culture of Candida halophila and Rhodotorula glutinis

In this study, the biomass production and pollutant removal from high-strength glutamate fermentation wastewater (GFW) using yeast isolates was investigated. Following enrichment culture, two species of yeasts, Candida halophila and Rhodotorula glutinis, were isolated from raw GFW with chemical oxygen demand (COD) and ammonia-nitrogen levels of 40 and 16 g l(-1), respectively. The binary mixed yeast culture was cultivated batchwise in 2.5-fold diluted GFW from which 85% of COD and 96% of reducing sugar were removed. The resulting yeast biomass contained 56% crude protein, 36.0% carbohydrate and 0.4% crude lipid. The amino acid composition of mixed yeast cells was balanced and was comparable with that of C. utilis and soybean.     

Evaluation of Simultaneous Nutrient and COD Removal with Polyhydroxybutyrate (PHB) Accumulation Using Mixed Microbial Consortia under Anoxic Condition and Their Bioinformatics Analysis

Simultaneous nitrate-N, phosphate and COD removal was evaluated from synthetic waste water using mixed microbial consortia in an anoxic environment under various initial carbon load (ICL) in a batch scale reactor system. Within 6 hours of incubation, enriched DNPAOs (Denitrifying Polyphosphate Accumulating Microorganisms) were able to remove maximum COD (87%) at 2g/L of ICL whereas maximum nitrate-N (97%) and phosphate (87%) removal along with PHB accumulation (49 mg/L) was achieved at 8 g/L of ICL. Exhaustion of nitrate-N, beyond 6 hours of incubation, had a detrimental effect on COD and phosphate removal rate. Fresh supply of nitrate-N to the reaction medium, beyond 6 hours, helped revive the removal rates of both COD and phosphate. Therefore, it was apparent that in spite of a high carbon load, maximum COD and nutrient removal can be maintained, with adequate nitrate-N availability. Denitrifying condition in the medium was evident from an increasing pH trend. PHB accumulation by the mixed culture was directly proportional to ICL; however the time taken for accumulation at higher ICL was more. Unlike conventional EBPR, PHB depletion did not support phosphate accumulation in this case. The unique aspect of all the batch studies were PHB accumulation was observed along with phosphate uptake and nitrate reduction under anoxic conditions. Bioinformatics analysis followed by pyrosequencing of the mixed culture DNA from the seed sludge revealed the dominance of denitrifying population, such as Corynebacterium, Rhodocyclus and Paraccocus (Alphaproteobacteria and Betaproteobacteria). Rarefaction curve indicated complete bacterial population and corresponding number of OTUs through sequence analysis. Chao1 and Shannon index (H’) was used to study the diversity of sampling. “UCI95” and “LCI95” indicated 95% confidence level of upper and lower values of Chao1 for each distance. Values of Chao1 index supported the results of rarefaction curve.     

Plant growth and nutrient removal in constructed monoculture and mixed wetlands related to stubble attributes

The aim of this study is to test the hypothesis that it depends on plant species used in the wetlands and their stubble growth attributes, as to whether monoculture or mixed wetland is superior in plant growth and nutrient removal. Monoculture and mixed wetland microcosms of five wetland plant species were studied. Significant differences in growth and aboveground biomass were found in the monoculture wetlands. Species that showed faster growth and larger biomass in monoculture wetland were also dominant in the mixed wetland. The mixed wetland exhibited similar biomass and root growth to the averages of five monocultures. ANOVA showed that there were very significant differences among the wetlands in removal rates of all the nutrients studied except nitrate nitrogen (NO₃-N) and chemical oxygen demand (COD). The removal rates from the mixed wetland were generally comparable to the highest removal rates from the monocultures. The species exhibited different stubble growth attributes, with some species showing increasing stubble growth and removal rates, while other species showing decreasing stubble growth and removal rates. The results indicated that in both monocultures and mixed constructed wetlands, growth and nutrient removal rates depended on plant species, and attributes of plant stubble growth affected overall growth and nutrient removal capabilities.     

Investigations on ultraviolet light and nitrous acid induced mutations of halotolerant bacterial strains for the treatment of tannery soak liquor

The objective of this research work is to study the effect of physical and chemical mutagenesis on biological treatment of tannery saline wastewater (soak liquor) employing halotolerant bacterial strains. Four halotolerant bacterial strains isolated from saline sources were used. The strains were identified as Pseudomonas aeruginosa, Bacillus flexus, Exiguobacterium homiense and Staphylococcus aureus, respectively. The isolates were found to grow well in medium containing 0–10% NaCl. At high saline concentration (>5%), the identified strains and their mixed consortia showed a low degrading efficiency of soak liquor (35–45%). UV light and nitrous acid mutagenesis were performed over the strains and the mutated strains were employed for degradation of soak liquor at high salinity level (6% by wt). Comparison of Chemical Oxygen Demand (COD) removal rates for both pure mutant isolates and mixed mutated consortia showed that nitrous acid mutagenesis resulted in degradation of 71% COD removal. Ultraviolet (UV) mutagenesis has no effect on degradation effectiveness. Biomass sludge (Mixed Liquor Volatile Suspended Solids) growth was also found to be high in nitrous acid treated strains.     

Kinetic model of a granular sludge SBR: influences on nutrient removal

A mathematical model was developed that can be used to describe an aerobic granular sludge reactor, fed with a defined influent, capable of simultaneously removing COD, nitrogen and phosphate in one sequencing batch reactor (SBR). The model described the experimental data from this complex system sufficiently. The effect of process parameters on the nutrient removal rates could therefore be reliably evaluated. The influence of oxygen concentration, temperature, granule diameter, sludge loading rate, and cycle configuration were analyzed. Oxygen penetration depth in combination with the position of the autotrophic biomass played a crucial role in the conversion rates of the different components and thus on overall nutrient removal efficiencies. The ratio between aerobic and anoxic volume in the granule strongly determines the N-removal efficiency as it was shown by model simulations with varying oxygen concentration, temperature, and granule size. The optimum granule diameter for maximum N- and P-removal in the standard case operating conditions (DO 2 mg L(-1), 20 degrees C) was found between 1.2 and 1.4 mm and the optimum COD loading rate was 1.9 kg COD m(-3) day(-1). When all ammonia is oxidized, oxygen diffuses to the core of the granule inhibiting the denitrification process. In order to optimize the process, anoxic phases can be implemented in the SBR-cycle configuration, leading to a more efficient overall N-removal. Phosphate removal efficiency mainly depends on the sludge age; if the SRT exceeds 30 days not enough biomass is removed from the system to keep effluent phosphate concentrations low.     

Repeated batch process for biological treatment of black liquor using brown-rot basidiomycete <Emphasis Type="Italic">Fomitopsis</Emphasis> sp. IMER2

A repeated batch operation is developed for the treatment of alkaline pulp black liquor, through a process of biological acidification precipitation of lignin using brown rot fungus Fomitopsis sp. IMER2. The results showed that COD and color removal of black liquor was dependent on the biomass concentration, pH decrease and initial COD. Based on these results, the repeated batch process was successfully carried out 12 times over 36 days in an air bubble column bioreactor. The average reduction of COD and color was approximately 40% and 70%, respectively.     

Enhanced biological nutrient removal using MUCT-MBR system

Biological nutrient removal was investigated in a combined modified University of Cape Town and membrane bioreactor system. When the influent nutrient mass ratio (COD/TN/TP) was 28.5/5.1/1 to 28.5/7.2/1, average removal efficiencies of COD, TN and TP were 90%, 81.6%, 75.2%. Obvious denitrifying phosphorus removal occurred with C/N ratio 3.98. When nitrite was the main electron acceptor, the ratio of denitrifying phosphate uptake to the total phosphate uptake were 99.8% and the sludge yield was 0.28kg VSS/kg COD; when nitrate was the main electron acceptor, the ratio was 92% and the yield was 0.32kg VSS/kg COD. In case of nitrite, the system not only kept TP and TN removal at 89.1% and 82.2%, but also ensured less sludge production. Batch tests showed that the proportion of denitrifying phosphorus-accumulating organisms in the total phosphorus-accumulating organisms in the system was higher than 80%.     

para-Chlorophenol inhibition on COD, nitrogen and phosphate removal from synthetic wastewater in a sequencing batch reactor

COD, nitrogen, phosphate and para-chlorophenol (4-chlorophenol, 4-CP) removal from synthetic wastewater was investigated using a four-step sequencing batch reactor (SBR) at different sludge ages and initial para-chlorophenol (4-CP) concentrations. The nutrient removal process consisted of anaerobic, oxic, anoxic and oxic phases with hydraulic residence times (HRT) of 1/3/1/1 h and a settling phase of 0.75 h. A Box-Wilson statistical experiment design was used considering the sludge age (5-25 days) and 4-CP concentration (0-400 mg l(-1)) as independent variables. Variations of percent COD, NH4-N, PO4-P and 4-CP removals with sludge age and initial 4-CP concentration were investigated. Percent nutrient removals increased with increasing sludge age and decreasing 4-CP concentrations. Low nutrient removals were obtained at high initial 4-CP concentrations especially at low sludge ages. However, high sludge ages partially overcome the adverse effects of 4-CP and resulted in high nutrient removals. COD, NH4-N, PO4-P and 4-CP removals were 76%, 72%, 26% and 34% at a sludge age of 25 days and initial 4-CP concentration of 200 mg l(-1). Sludge volume index (SVI) also decreased with increasing sludge age and decreasing 4-CP concentrations. An SVI value of 104 ml g(-1) was obtained at a sludge age of 25 days and initial 4-CP of 200 mg l(-1).     

Simultaneous COD, nitrogen, and phosphate removal by aerobic granular sludge

Aerobic granular sludge technology offers a possibility to design compact wastewater treatment plants based on simultaneous chemical oxygen demand (COD), nitrogen and phosphate removal in one sequencing batch reactor. In earlier studies, it was shown that aerobic granules, cultivated with an aerobic pulse-feeding pattern, were not stable at low dissolved oxygen concentrations. Selection for slow-growing organisms such as phosphate-accumulating organisms (PAO) was shown to be a measure for improved granule stability, particularly at low oxygen concentrations. Moreover, this allows long feeding periods needed for economically feasible full-scale applications. Simultaneous nutrient removal was possible, because of heterotrophic growth inside the granules (denitrifying PAO). At low oxygen saturation (20%) high removal efficiencies were obtained; 100% COD removal, 94% phosphate (P-) removal and 94% total nitrogen (N-) removal (with 100% ammonium removal). Experimental results strongly suggest that P-removal occurs partly by (biologically induced) precipitation. Monitoring the laboratory scale reactors for a long period showed that N-removal efficiency highly depends on the diameter of the granules.     

Effect of nickel on nutrient removal by selected indigenous protozoan species in wastewater systems

Nutrient and heavy metal pollutions are major concern worldwide. This study aimed at comparing the effect of Ni²⁺ on nutrient removal efficiency of four indigenous wastewater protozoan species (Aspidisca sp., Paramecium sp., Peranema sp., Trachelophyllum sp.). Specific physicochemical parameters and microbial growth/die-off were measured using standard methods. The results revealed that protozoan species were able to simultaneously remove phosphate, nitrate and Ni²⁺ at concentrations ranging between 66.4–99.36%, 56.19–99.88% and 45.98–85.69%, respectively. Peranema sp. appeared to be the isolates with the highest removal of nutrients (Phosphate-99.36% and Nitrate-99.88%) while Paramecium sp. showed higher removal of Ni²⁺ at 85.69% and low removal of nutrients. Aspidisca sp. was the most sensitive isolate to Ni²⁺ but with significant nutrient removal (Phosphate-66.4% and Nitrate-56.19%) at 10 mg-N²⁺/L followed by an inhibition of nutrient removal at Ni²⁺ concentration greater than 10 mg/L. Significant correlation between the growth rate and nutrient removal (r = 0.806/0.799, p < 0.05 for phosphate and nitrate, respectively) was noted. Except for Peranema sp. which revealed better nutrient removal ability at 10 mg-Ni²⁺/L, an increase in Ni²⁺ concentration had a significant effect on nutrient removal efficiency of these indigenous protozoan species. This study suggests that although Ni²⁺ appeared to be toxic to microbial isolates, its effect at a low concentration (10 mg-Ni²⁺/L) towards these isolates can be used to enhance the wastewater treatment process for the removal of nutrients. Peranema sp., which was able to remove both Ni²⁺ and nutrients from wastewater mixed-liquor, can also be used for bioremediation of wastewater systems.     

Converting corn wet-milling effluent into high-value fungal biomass in a biofilm reactor

Rhizopus microsporus was grown in an attached growth system using corn wet-milling effluent as a growth medium. This strain was chosen due to its ability to effectively degrade organic matter in corn wet-milling effluent and for its properties to produce significant levels of protein, chitin and chitosan. Fungal growth and organic removal efficiency were examined under both aseptic and non-aseptic conditions with and without nutrient supplementation. Plastic composite support (PCS) tubes, composed of 50% (w/w) polypropylene (PP) and 50% (w/w) agricultural products were used as support media. Significantly higher organic removal measured as chemical oxygen demand (COD) and biomass yield were observed in the bioreactor with PCS tubes than in two control bioreactors; that is with PP tubes alone and suspended growth (without support media). This confirmed that the PCS support medium with agricultural components enhanced fungal growth and organic removal. The results showed that supplementation of nutrients (e.g., mineral salts) under aseptic conditions enhanced the COD removal from 50% to 55% and observed biomass yield from 0.11 to 0.16 g (dry-weight)/g COD(removed) (i.e., from 0.10 to 0.14 g volatile solids (VS)/g COD(removed) approximately). Non-aseptic operation without nutrient supplementation resulted in an observed biomass yield of 0.32 g volatile suspended solids (VSS)/g COD(removed) with no significant improvement in COD removal ( approximately 53%); whereas with nutrient supplementation, the observed biomass yield increased to 0.56 g VSS/g COD(removed) and COD removal improved to 85%. The fungal system was able to degrade the organic matter with concomitant production of high-value fungal biomass. This is the first study that examined the conversion of corn milling waste stream into high value fungal protein.     

Enhanced remediation of black liquor by activated sludge bioaugmented with a novel exogenous microorganism culture

Black liquor (BL) is a notoriously difficult wastewater to treat due to the economic and efficiency limitations of physiochemical methods and intrinsic difficulties with bioremediation strategies caused by the high pH (10–13) and lignin content. This study investigated the feasibility of a novel bioaugmentation strategy for BL treatment, which uses a mixed microorganism culture of lignocellulose-degrading microorganisms isolated from degraded bamboo slips. Black liquor treatment was assessed in terms of chemical oxygen demand (COD) and color removal with a sequencing batch reactor organic loading rate of 9 kg COD/L·day under highly alkaline conditions (pH?10). Results revealed that bioaugmented activated sludge treatment of BL with special mixed microorganisms significantly enhanced the removal efficiency of COD, color, and lignin from the wastewater up to 64.8, 50.5, and 53.2 %, respectively. Gel permeation chromatography profiles showed that the bioaugmentation system could successfully degrade high molecular lignin fragments in black liquor. This work confirms bioaugmentation as a feasible alternative strategy for enhanced biological treatment of wastewater with high lignin content and high organic load rate under strongly alkaline conditions.     

Improved algal oil production from Botryococcus braunii by feeding nitrate and phosphate in an airlift bioreactor

To improve biomass and microalgal oil production of Botryococcus braunii, fed‐batch culture was investigated in an airlift photobioreactor. The optimal feeding time of the fed‐batch culture was after 15 days of cultivation, where 1.82 g/L of the microalgal biomass was obtained in the batch culture. Nitrate nutrient was the restrictive factor for the fed‐batch cultivation while phosphate nutrient with high concentration did not affect the microalgal growth. The optimal mole ratio of nitrate to phosphate was 34.7:1, where nitrate concentration reached the initial level and phosphate concentration was one quarter of its initial level. With one feeding, the biomass of B. braunii reached 2.56 g/L after 18 days. Two feedings in 2‐day interval enhanced the biomass production up to 2.87 g/L after 19 days of cultivation. The hydrocarbon content in dry biomass of B. braunii kept at high level of 64.3% w/w. Compared with the batch culture, biomass production and hydrocarbon productivity of B. braunii were greatly improved by the strategic fed‐batch cultivation.     

Comparison of the performance of submerged membrane bioreactor (SMBR) and submerged membrane adsorption bioreactor (SMABR)

This study focuses on comparing the performance of submerged membrane bioreactor (SMBR) and submerged membrane adsorption bioreactor (SMABR) over a period of 20 days at a hydraulic retention time (HRT) of 3.1h. The effects of PAC on critical flux and membrane fouling were also investigated. The SMABR exhibited better results in terms of mixed liquor suspended solids (MLSS) growth, DOC removal (over 96%), COD removal (over 95%), transmembrane pressure (TMP) and oxygen uptake rate. Nearly 100% of bacteria and 100% of total coliforms were removed in both systems. The addition of PAC could maintain the critical flux at a lower TMP value (7.5 kPa), while irreversible fouling caused by PAC occurred when the filtration flux exceeded critical flux.     

Combined biological and membrane treatment of food-processing wastewater to achieve dry-ditch criteria: pilot and full-scale performance

This study tested the applicability of a submerged vacuum ultrafiltration membrane technology in combination with the biological treatment system to achieve dry-ditch criteria stipulated as follows: BOD5, TSS, NH3-N, and total phosphorous (TP) concentration not exceeding 10, 10, 1, and 0.5 mg/L respectively for the treatment of high strength food-processing wastewater. During the study, the biological system, operated at average hydraulic retention time of 5-6 days, achieved 95-96.5% BOD removal and 96-99% COD removal. The external membrane system ensured the achievability of the BOD and TSS criteria, with BOD and TSS concentrations in the permeate of 1-2 and 1-8 mg/L respectively. Nitrate, and nitrite concentrations increased during membrane filtration, while ammonia concentrations decreased. The most salient finding of this study is that, contrary to common belief, for industrial wastewaters, the filterability of the mixed liquor is influenced by the soluble organics, and may be low, thus necessitating operation of bioreactors at low mixed liquor solids. This study demonstrated that bioreactors operated at low SRTs and in combination with ultrafiltration can still achieve superior effluent quality that may meet reuse criteria at reasonable cost.     

Enhanced biological phosphate removal by granular sludge in a sequencing batch reactor

A laboratory-scale sequencing batch reactor was started-up with flocculated biomass and operated primarily for enhanced biological phosphate removal. Ten weeks after the start-up, gradual formation of granular sludge was observed. The compact biomass structure allowed halving the settling time, the initial reactor volume, and doubling the influent COD concentration. Continued operation confirmed the possibility of maintaining a stable granular biomass with a sludge volume index less than 40 ml g^–1, while securing a removal efficiency of 95% for carbon, 99.6% for phosphate, and 71% for nitrogen. Microscopic observations revealed a morphological diversity.     

Impact of fly ash pretreatment on aerobic treatment of thermomechanical pulping spent liquor

This study investigated the impact of biomass‐based fly ash (FA) pretreatment on the biodegradability of a thermomechanical pulping spent liquor (TMPL) in an aerobic system. In this study, FA was mixed with TMPL under the conditions of 6 wt.% based on TMPL, 25°C and 10 h, which removed a part of recalcitrant organic materials and resulted in 68.0, 40.0, 60.1, 81.2 and 48.3% reductions in chemical oxygen demand (COD), biochemical oxygen demand (BOD), total organic carbon (TOC), lignin and sugar, respectively. FA‐pretreated TMPL pressate (FA‐TMPL) was biologically treated in an aerobic system of sequencing batch reactor (SBR). The performance of the biological treatment with and without FA pretreatment was studied in two parallel SBRs over three months. The combination of FA and biological treatments removed 97.3% of COD, 98.3% of BOD, 96.3% of lignin, 99.5% of sugar, and 98.1% of TOC. Without FA pretreatment, the biological system removed 87.3% of COD, 89% of BOD, 81.6% of lignin, 98.6% of sugars, and 90.5% of TOC. The results also confirmed that the settling ability of sludge, which was indicated as a sludge volume index, was reduced from 109.3 mL/g to 53.5 mL/g. In addition, the advantages of using FA pretreatment in aerobic systems were discussed in detail. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:370–378, 2018     

Removal of organic pollutants and analysis of MLSS–COD removal relationship at different HRTs in a submerged membrane bioreactor

In order to investigate the influence of hydraulic retention time (HRT) on organic pollutant removal in a submerged membrane bioreactor (SMBR), a laboratory-scale experiment was conducted using domestic sewage as influent. The dissolved oxygen (DO) concentration was controlled at 2.0– during the experimental period. The experiments demonstrated that when HRT was 3, 2 and 1 h, the reduction of chemical oxygen demand (COD) was 89.3–97.2, 88.5–97.3 and 80–91.1%, and the effluent COD was 38.9–11.2, 41.6–10.8 and 63.4–, respectively. It is suggested that an HRT of 1 h could meet the normal standard of discharged domestic sewage, and an HRT of 2 h could meet that of water reclamation. In addition, we use mathematical software MATLAB to analyse the relation of mixed liquor suspended solids (MLSS) and COD removal. The results showed that the optimum MLSS concentration should be maintained at around in the SMBR. The results also showed that the COD removal was related to HRT (τ), influent concentration (S₀) and sludge loading rate for COD removal (N_S). Moreover, the high COD removal could be achieved through adjusting τ, S₀ and N_S.