Regeneration of oxidative assimilation capacity by intracellular conversion of storage products to protein

Previous studies have shown that the capacity of a heterogeneous microbial population for oxidative assimilation of glucose can be renewed by periodically subjecting the sludge (or a portion of it) to endogenous respiration in the presence of an exogenous source of nitrogen. Further study of this system led to a modification of the activated sludge process for nitrogen-deficient wastes. However, it was not known whether renewal of oxidative assimilation capacity was possible for substrates which required the presence of inducible enzyme(s) or for substrates which were not carbohydrates. Therefore, studies with lactose and acetate as carbon sources were designed. Both carbon sources were removed under conditions of oxidative assimilation, and the storage products (or a portion of these products) were converted into protein when the sludge was subjected to a period of endogenous respiration (with respect to carbon source) in the presence of an exogenous supply of ammonium sulfate. The “regenerated” sludge exhibited a renewed capacity for oxidative assimilation, thereby indicating that requisite inducible enzymes (e.g., β-galactosidase in the case of lactose; iso-citritase and malate synthetase in the case of acetate) were not diluted out in the endogenous phase to a degree sufficient to hamper renewed oxidative assimilation capacity. The results also indicated that a noncarbohydrate carbon source can be successfully removed from the medium with this process. However, in the case of acetate, the oxidative assimilation capacity after “regeneration” was not fully restored to the initial level.     

Oxidative assimilation treatment of a nitrogen-deficient toxic waste

Batch growth tests were performed under both replicating and nonproliferating (no nitrogen source in medium) conditions with acclimated heterogenous populations that utilized phenol as a sole source of carbon and energy. It was shown that the acclimated populations could efficiently remove the toxic waste component phenol under nonproliferating conditions by utilizing an oxidative assimilation mechanism. The phenol was assimilated and converted into nonnitrogenous storage products. During the assimilation process, the cells had a tendency to excrete some product (nonsubstrate) chemical oxygen demand (COD). Bench-scale oxidative assimilation units were operated by sequentially feeding a carbon source (phenol) and nitrogen source (ammonium sulfate) to heterogeneous populations. This demonstrated that, subsequent to the addition of the nitrogen source to the medium, the cells utilized the stored carbon for replication. Four of these units were operated at different phenol COD-to-ammonia-nitrogen ratios of 10:1, 20:1, 40:1, and 50:1. All of these units demonstrated excellent removal of phenol using an oxidative assimilation mechanism. These results suggested the feasibility of utilizing a continuous flow oxidative assimilation process for the treatment of nitrogen-deficient phenolic wastes. This process would be advantageous over conventional treatment processes in that it would realize a savings in chemical costs (ammonia as nitrogen source) and prevent leakage of excess ammonia from the system.     

Continuous oxidative assimilation of acetic Acid and endogenous protein synthesis applicable to treatment of nitrogen-deficient waste waters

A previous study indicated that the oxidative assimilation capacity of a heterogeneous microbial population for acetic acid did not return to the initial value after a period of endogenous respiration in the presence of ammonium sulfate. In view of this finding it was not possible to recommend fully the use of a continuous oxidative assimilation process for the treatment of nitrogen-deficient waste waters of noncarbohydrate nature. To put the process to a severe test, a laboratory-scale pilot plant study was done. Acetate (1,000 mg/liter) was fed continuously to a completely mixed, aerated reactor vessel, from which the mixed liquor was channeled to a settling basin. The settling basin supernatant fluid was continuously discharged, and a portion of the settled biological sludge was subjected to endogenous respiration in the presence of ammonium sulfate and was recycled to the aeration vessel. Experiments were conducted at three levels of ammonia nitrogen in the endogenous phase. Approximately 90% removal of the carbon source (expressed as chemical oxygen demand, COD) was attained with a low level of supplemental nitrogen (COD/N = 70:1) and a fairly low reactor detention time (4 hr). Based upon these and previous results, it has been concluded that the process shows promise for application to the treatment of a wide variety of nitrogen-deficient industrial wastes.     

Carbon source recovery from waste activated sludge by alkaline hydrolysis and gamma-ray irradiation for biological denitrification

The recovery of an organic carbon source from a waste activated sludge by using alkaline hydrolysis and radiation treatment was studied, and the feasibility of the solubilized sludge carbon source for a biological denitrification was also investigated. The effects of an alkaline treatment and gamma-ray irradiation on a biodegradability enhancement of the sludge were also studied. A modified continuous bioreactor for a denitrification (MLE reactor) was operated by using a synthetic wastewater for 47 days. Alkaline treatment of pH 10 and gamma-ray irradiation of 20 kGy were found to be the optimum carbon source recovery conditions. COD removal of 84% and T-N removal of 51% could be obtained by using the solubilized sludge carbon source through the MLE denitrification process. It can be concluded that the carbon source recovered from the waste activated sludge was successfully employed as an alternative carbon source for a biological denitrification.     

Managing the excessive proliferation of glycogen accumulating organisms in industrial activated sludge by nitrogen supplementation: A FISH-NanoSIMS approach

Defluviicoccus vanus-related glycogen accumulating organisms (GAO) regularly proliferate in industrial wastewater treatment plants handling high carbon but nitrogen deficient wastes. When GAO dominate, they are associated with poor performance, characterised by slow settling biomass and turbid effluents. Although their ecophysiology has been studied thoroughly in domestic waste treatment plants, little attention has been paid to them in aerobic industrial systems.In this study, the effect of nitrogen addition on GAO carbon metabolism was investigated during an 8 h cycle. Activated sludge dominated by GAO from a winery wastewater sequencing batch reactor was incubated under different carbon to nitrogen (COD:N) ratios (100:1, 60:1 and 20:1) using ¹³C — acetate and ¹⁵N — urea. GAO cell assimilation was quantified using FISH-NanoSIMS. The activated sludge community was assessed by 16S rRNA gene profiling, DNA and storage polymer production. Carbon and nitrogen quantification at the cellular level by NanoSIMS revealed that low (COD:N of 100:1) or null nitrogen concentrations enhanced GAO carbon uptake. COD:N ratios of 60:1 and 20:1 reduced GAO carbon uptake and promoted whole microbial community DNA production. Nitrogen dosing at COD:N ratios of 60:1 or higher was demonstrated as feasible strategy for controlling the excessive GAO growth in high COD waste treatment plants.     

The investigation of effect of organic carbon sources addition in anaerobic–aerobic (low dissolved oxygen) sequencing batch reactor for nutrients removal from wastewaters

The effect of addition of organic carbon sources (acetic acid and waste activated sludge alkaline fermentation liquid) on anaerobic–aerobic (low dissolved oxygen, 0.15–0.45 mg/L) biological municipal wastewater treatment was investigated. The results showed that carbon source addition affected not only the transformations of polyhydroxyalkanoates (PHA), glycogen, nitrogen and phosphorus, but the net removal of nitrogen and phosphorus. The removal efficiencies of TN and TP were, respectively, 61% and 61% without organic carbon source addition, 81% and 95% with acetic acid addition, and 83% and 97% with waste activated sludge alkaline fermentation liquid addition. It seems that the alkaline fermentation liquid of waste biosolids generated in biological wastewater treatment plant can be used to replace acetic acid as an additional carbon source to improve the anaerobic–aerobic (low dissolved oxygen) municipal wastewater nutrients removal although its use was observed to cause a slight increase of effluent BOD and COD concentrations.     

Einfluß des Phasenstatus der Vorkulturzellen auf die Ascosporenbildung von Saccharomyces cerevisiae

Summary Cells from three growth phases were examined for their ability to sporulate: cells from a) phase II (first phase of exponential growth with glucose as carbon source), b) phase III (second lag-phase during adaptation to oxidative metabolism), and c) phase IV (second phase of almost exponential growth with ethanol as carbon source). 1. Cells from phase III showed the best sporulation ability because they reached the highest percentage of asci and also of 4-spored asci. 2. Cells of phase II exhibited the highest and those of phase IV the lowest rate of sporulation (Fig. 3). 3. The longer the cells remained in the presporulation medium the more abbreviated was the time in the sporulation culture before the first asci appeared, and this abbreviation was just equal to the time of elongation in the preculture. This clearly demonstrates the different degree of respiratory adaptation. — After transfer to the sporulation medium O₂-consumption arose to a steep maximum within the first 10 hours followed by medium values which dropped again rapidly at the onset of ascospore formation (Fig. 4). Only during the time of high and medium O₂-consumption there was an increase in dry weight reflecting the assimilation of acetate. In cells of phase II compared with those of phase IV this assimilation of acetate showed the same delay as the onset of sporulation, whereas full capacity of respiration was reached much sooner.     

Composting potential of different inoculum sources in the modified SEBAC system treatment of municipal solid wastes

The aim of the work described here was to analyse the biomethanization process for three types of organic fraction of municipal solid waste (OFMSW) using two different inoculum sources in a sequential leach-bed anaerobic reactor under thermophilic (55 degrees C) and dry conditions (20%TS). The OFMSWs studied were: separately collected food waste (SC_OFMSW) from restaurants, synthetic waste (ST_OFMSW) and mechanically selected municipal waste (MS_OFMSW). The inoculum studied were digested mesophilic sludge (SLUDGE) and digested SC_OFMSW. The results show that SLUDGE gave the best performance and this was therefore selected for the rest of the experiments. Three assays were performed to analyse the biomethanization processes of three types of organic waste. The results suggest that all three wastes give rise to an acclimation stage with acidogenic/acetogenic activity between days 5 and 20-30 and a stabilization phase associated with methanogenic activity. In conclusion, a mixture of OFMSW (regardless of its origin) and SWINE arranged in layers in the reactor, as well as SLUDGE used with an inoculum source, enhanced the fast start up phase of a modified sequential leach-bed system under dry thermophilic conditions.     

Microbial sulfate reduction at low (8 °C) temperature using waste sludge as a carbon and seed source

Biological treatment of sulfate and metal-containing wastewater (such as acid mine drainage) is a viable option due to lower cost and better sludge quality compared to conventional chemical treatment. Although several substrates can be used as carbon source, a low-cost substrate is required for large scale applications. This study was conducted to investigate the suitability of waste sludge as a carbon and seed source for sulfate reduction at 8 °C in batch bioassays. Around 7 mmol of sulfate was reduced when the waste sludge mixture (WS) (6700 mg SS l^?1) from primary and secondary settling tank was supplemented as a carbon and seed source. However, only 1.6 mmol of sulfate was reduced with anaerobic digester effluent (ADS) (5300 mg SS l^?1). The produced H₂S from 1 g VSS l^?1 WS and ADS oxidation can theoretically precipitate around 90 and 35 mg Fe²⁺, respectively. Both WS and ADS oxidized ethanol to acetate at similar rates. It appears that WS is a good candidate for carbon and start-up seed source of sulfate reduction at 8 °C, whereas sulfidogenic acetate oxidation was the limiting step. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed that both sludge sources contain Desulfomicrobium apsheronum strain.     

Interrelations between glycogen,poly-β-hydroxybutyric acid and lipids during accumulation and subsequent utilization in a Pseudomonas

A pleomorphic pseudomonad, V-19, was isolated from activated sludge on the basis of its floc-forming capacity in 0.1% casitone ?0.035% yeast extract (CYE) ?0.2% glycerol medium. In the late exponential phase of growth morphological changes and flocculation phenomena took place accompanied by a massive deposition of reserve granules in the cell. Chemical and electron-microscopical examination revealed 3 types of storage products: glycogen, poly-β-hydroxybutyric acid (PHB) and ether-extractable lipids. These products were isolated and chemically characterized. In CYE medium supplied with 0.5% glucose or glycerol as the carbon source mainly ether-soluble lipids and glycogen were synthesized. On continued incubation these materials were slowly utilized, which enabled the cells to survive for long periods of time. Growth in inorganic salts medium (0.1% ammonium sulfate; 1% carbon source) yielded cells containing different accumulated products, depending on the carbon source used. Glycerol-grown cells contained mainly glycogen, but also ether-soluble lipid, and no PHB. Glucose was largely converted into gluconic acid and excreted into the medium before being deposited in the form of PHB as the primary product of assimilation. Subsequently, PHB was metabolized thereby being partly transformed into glycogen and ether-soluble lipid. Addition of ammonium sulfate to nitrogen-starved cells caused a ready mobilization of the accumulated products, resulting in a net synthesis of reservefree cell material and an increase in the number of cells.     

Anaerobic co-digestion of algal sludge and waste paper to produce methane

The unbalanced nutrients of algal sludge (low C/N ratio) were regarded as an important limitation factor to anaerobic digestion process. Adding high carbon content of waste paper in algal sludge feedstock to have a balanced C/N ratio was undertaken in this study. The results showed adding 50% (based on volatile solid) of waste paper in algal sludge feedstock increased the methane production rate to 1170+/-75 ml/l day, as compared to 573+/-28 ml/l day of algal sludge digestion alone, both operated at 4 g VS/l day, 35 degrees C and 10 days HRT. The maximum methane production rate of 1607+/-17 ml/l day was observed at a combined 5 g VS/l day loading rate with 60% (VS based) of paper adding in algal sludge feedstock. Results suggested an optimum C/N ratio for co-digestion of algal sludge and waste paper was in the range of 20-25/1.     

Methanol‐driven enhanced biological phosphorus removal with a syntrophic consortium

The presence of suitable carbon sources for enhanced biological phosphorus removal (EBPR) plays a key role in phosphorus removal from wastewater in urban WWTP. For wastewaters with low volatile fatty acids (VFAs) content, an external carbon addition is necessary. As methanol is the most commonly external carbon source used for denitrification it could be a priori a promising alternative, but previous attempts to use it for EBPR have failed. This study is the first successful report of methanol utilization as external carbon source for EBPR. Since a direct replacement strategy (i.e., supply of methanol as a sole carbon source to a propionic‐fed PAO‐enriched sludge) failed, a novel process was designed and implemented successfully: development of a consortium with anaerobic biomass and polyphosphate accumulating organisms (PAOs). Methanol‐degrading acetogens were (i) selected against other anaerobic methanol degraders from an anaerobic sludge; (ii) subjected to conventional EBPR conditions (anaerobic + aerobic); and (iii) bioaugmented with PAOs. EBPR with methanol as a sole carbon source was sustained in a mid‐term basis with this procedure. Biotechnol. Bioeng. 2013; 110: 391–400. © 2012 Wiley Periodicals, Inc.     

Photo-Fenton process for excess sludge disintegration

For the minimization of excess sludge in activated sludge process, the photo-Fenton reaction was applied as a novel technique and its effectiveness was experimentally examined. A batch study was conducted to elucidate parameters governing the activated sludge integration by the photo-Fenton process such as the concentrations of sludge, Fe ion and hydrogen peroxide. It was found that the chemical oxidation sludge disintegration by photo-Fenton reaction could be divided into two phases. At the beginning of the photo-Fenton process the dissolved chemical oxygen demand (COD) increased. This is due to the discharge of organic compounds occurred by oxidative decomposition of the cell wall of a microbe in the sludge. The COD reached the maximum and then decreased. In this phase, mineralization of dissolved organic substances by the photo-Fenton reaction might be dominant. The occurrence of mineralization was suppressed as the initial MLSS increased. The Fe dosage enhanced the sludge disintegration by the photo-Fenton reaction as well as the H₂O₂ dosage. The soluble total organic carbon (TOC) of the solution increased monotonously. The results suggest that Fe ions might be entrapped into the activated sludge. It may be concluded that photo-Fenton reaction is one of feasible processes for disintegration of excess activated sludge.     

Removal capacity of water plant alum sludge for phosphorus in aqueous solutions

Abstract

Alum sludge, which is a waste product from a potable water treatment process, was tested as an inexpensive alternate adsorbent for phosphorus in wastewater. The sludge was composed dominantly of sand size aggregates, and could remain stable in aqueous media. The majority of reactive Al in alum sludge was present as an amorphous phase, and seemed to be the major absorbent for P. The batch sorption test showed that the removal of P was influenced by the solubility of Al, Fe and organic carbon depending on pH condition. The acidic condition favored the removal of P, and there was a side effect in the P removal process such as dissolution of Al and organic C at acidic (pH < 4) and alkaline (pH < 8) conditions. The pH range from 4 to 6 was effective for all inorganic and organic phosphates with a low solubility of Al and organic C. The maximum adsorption capacity of alum sludge was calculated as 25,000 mg/kg for orthophosphate, and followed the order: orthophosphate > pyrophosphate > triphosphate > organic phosphate (adenosin). From the column test with a 30 mg/L orthophosphate solution at a flow rate of 3.0 ml/min, the alum sludge removed P to less than 1.0 mg/L over 250 pore volumes at initial pH 4, and 200 pore volumes at initial pH 5, respectively.     

Microscopic observation of aerobic granulation in sequential aerobic sludge blanket reactor

AIMS: This paper attempts to provide visual evidence of how aerobic granulation evolves in sequential aerobic sludge blanket reactors. METHODS AND RESULTS: A series of experiments were conducted in two column-type sequential aerobic sludge reactors fed with glucose and acetate as sole carbon source, respectively. The evolution of aerobic granulation was monitored using image analysis and optical and scanning electron microscopy. The results indicated that the formation of aerobic granules was a gradual process from seed sludge to compact aggregates, further to granular sludge and finally to mature granules with the sequential operation proceeding. Glucose- and acetate-fed granules have comparable characteristics in terms of settling velocity, size, shape, biomass density and microbial activity. However, the microbial diversity of the granules was associated with the carbon source supplied. In this work, an important aerobic starvation phase was identified during sequential operation cycles. It was found that periodical aerobic starvation was an effective trigger for microbial aggregation in the reactor and further strengthened cell-cell interaction to form dense aggregates, which was an essential step of granulation. The periodical starvation-induced aggregates would finally be shaped to granules by hydrodynamic shear and flow. CONCLUSION: Aerobic granules can be formed within 3 weeks in the systems. The periodical starvation and hydrodynamic conditions would play a crucial role in the granulation process. SIGNIFICANCE AND IMPACT OF THE STUDY: Aerobic granules have excellent physical characteristics as compared with conventional activated sludge flocs. This research could be helpful for the development of an aerobic granule-based novel type of reactor for handling high strength organic wastewater.     

Free-living dinitrogen-fixing bacteria isolated from petroleum refinery oily sludge

Dinitrogen-fixing activity (acetylene reduction and N(2) fixation) was found in an oily sludge originating from a petroleum refinery. Two representative dinitrogen-fixing bacterial strains were isolated from this oily waste. Their nitrogenase activity was effective when they were cultivated on sterilized sludge or simple carbon substrates (organic acid salts, sugars). Using the classical methods, these strains could not be unambiguously related to other diazotrophic taxa. The landfarming process is widely used for oily sludge disposal; this study shows that oily sludges are more than a simple carbon input into the soil but that they must also be considered as real sources of dinitrogen-fixing and probably degradative microorganisms.     

Pyrolysis of plant, animal and human waste: physical and chemical characterization of the pyrolytic products

Pyrolysis (carbonization) has been proposed as one of several optional technologies for disposing and recycling waste products in Japan. Plant wastes (sugarcane bagasse and rice husks), animal waste (cow biosolids) and human waste (treated municipal sludge) were pyrolyzed at temperatures from 250–800 °C in closed containers. The carbonized materials were evaluated for specific physical properties (yield, surface area, density) and specific chemical properties (total carbon, total nitrogen, pH, fixed carbon, ash content, volatility) in order to compare differences in properties among the four waste products. The results indicated that (1) surface area, total carbon, ash content and pH increased as the carbonization temperature increased, while carbonization yield decreased with increasing temperature, (2) product density however was not affected by temperature and (3) correlation coefficients were determined among the physical and chemical properties and several significant correlations were observed. The data indicate that source material had considerable influence on the physical and chemical properties of the carbonized products.     

Influence of total solid and inoculum contents on performance of anaerobic reactors treating food waste

The aim of this paper was to analyze the biomethanization process of food waste (FW) from a university campus restaurant in six reactors with three different total solid percentages (20%, 25% and 30% TS) and two different inoculum percentages (20-30% of mesophilic sludge). The experimental procedure was programmed to select the initial performance parameters (total solid and inoculum contents) in a lab-reactor with V: 1100mL and, later, to validate the optimal parameters in a lab-scale batch reactor with V: 5000mL. The best performance for food waste biodegradation and methane generation was the reactor with 20% of total solid and 30% of inoculum: give rise to an acclimation stage with acidogenic/acetogenic activity between 20 and 60 days and methane yield of 0.49L CH4/g VS. Also, lab-scale batch reactor (V: 5000mL) exhibit the classical waste decomposition pattern and the process was completed with high values of methane yield (0.22L CH4/g VS). Finally, a protocol was proposed to enhance the start-up phase for dry thermophilic anaerobic digestion of food waste.     

Dry-thermophilic anaerobic digestion of organic fraction of the municipal solid waste: focusing on the inoculum sources

The effect of inoculum source on anaerobic thermophilic digestion of separately collected organic fraction of municipal solid wastes (SC_OFMSW) has been studied. Performance of laboratory scale reactors (V: 1.1 L) were evaluated using six different inoculums sources: (1) corn silage (CS); (2) restaurant waste digested mixed with rice hulls (RH_OFMSW); (3) cattle excrement (CATTLE); (4) swine excrement (SWINE); (5) digested sludge (SLUDGE); and (6) SWINE mixed with SLUDGE (1:1) (SWINE/SLUDGE). The SC_OFMSW was separately and collected from university restaurant. The selected conditions were: 25% of inoculum, 30% of total solid and 55 degrees C of temperature, optimum in the thermophilic range. The six inoculum sources showed an initial start-up phase in the range between 2 and 4 days and the initial methane generation began over 10 days operational process. Results indicated that SLUDGE is the best inoculum source for anaerobic thermophilic digestion of the treatment of organic fraction of municipal solid waste at dry conditions (30%TS). Over 60 days operating period, it was confirmed that SLUDGE reactor can achieve 44.0%COD removal efficiency and 43.0%VS removal. In stabilization phase, SLUDGE reactor showed higher volumetric biogas generated of 78.9 mL/day (or 35.6 mLCH(4)/day) reaching a methane yield of 0.53 LCH(4)/gVS. Also, SWINE/SLUDGE and SWINE were good inoculums at these experimental conditions.     

Flux balance analysis of Chlorella sp. FC2 IITG under photoautotrophic and heterotrophic growth conditions

Quantification of carbon flux distribution in the metabolic network of microalgae remains important to understand the complex interplay between energy metabolism, carbon fixation, and assimilation pathways. This is even more relevant with respect to cyclic metabolism of microalgae under light–dark cycle. In the present study, flux balance analysis (FBA) was carried out for an indigenous isolate Chlorella sp. FC2 IITG under photoautotrophic and heterotrophic growth conditions. A shift in intracellular flux distribution was predicted during transition from nutrient sufficient phase to nutrient starvation phase of growth. Further, dynamic flux analysis (dFBA) was carried out to capture light–dark metabolism over discretized pseudo steady state time intervals. Our key findings include the following: (i) unlike heterotrophic condition, oxidative pentose phosphate (PP) pathway, and Krebs cycle were relatively inactive under photoautotrophic growth; (ii) in both growth conditions, while transhydrogenation reaction was highly active, glyoxalate shunt was found to be nonoperative; (iii) flux distribution during transition period was marked with up regulation of carbon flux toward nongrowth associated (NGA) maintenance energy, oxidative phosphorylation, and photophosphorylation; (iv) redirection of carbon flux from polysaccharide and neutral lipid resulted in up regulation of Krebs cycle flux in the dark phase; (v) elevated glycolytic and acetyl-CoA flux were coupled with induction of neutral lipid during light cycle of the growth; (vi) significantly active photophosphorylation in the light phase was able to satisfy cellular energy requirement without need of oxidative PP pathway; and (vi) unlike static FBA, dFBA predicted an unaltered NGA maintenance energy of 1.5 mmol g^?1 DCW h^?1.