Ultrasonic treatment of biological sludge: Floc disintegration, cell lysis and inactivation

The aim of this work was to study the effect of ultra sound treatment on the solid content of sludge and biological activity, and the increase in the soluble chemical oxygen demand (SCOD), proteins and nucleic acids concentrations during sonication. The results showed that sonication effectively degraded and inactivated the sludge. The sludge disintegration and cell lysis occurred continuously while sludge inactivation mainly occurred in the second stage (10-30 min) during sonication. The SCOD, supernatant proteins and nucleic acids concentrations, and sludge mass reduction and inactivation degrees increased with the sonication time and power density increases. Higher energy ultrasound was more efficient than lower energy ultrasound for the sludge treatment.     

Entomotoxicity,protease and chitinase activity of Bacillus thuringiensis fermented wastewater sludge with a high solids content

This study investigated the production of biopesticides, protease and chitinase activity by Bacillus thuringiensis grown in raw wastewater sludge at high solids concentration (30 g/L). The rheology of wastewater sludge was modified with addition of Tween-80 (0.2% v/v). This addition resulted in 1.6 and 1.3-fold increase in cell and spore count, respectively. The maximum specific growth rate (μ_max) augmented from 0.17 to 0.22 h⁻¹ and entomotoxicity (Tx) increased by 29.7%. Meanwhile, volumetric mass transfer coefficient (k_La) showed marked variations during fermentation, and oxygen uptake rate (OUR) increased 2-fold. The proteolytic activity increased while chitinase decreased for Tween amended wastewater sludge, but the entomotoxicity increased. The specific entomotoxicity followed power law when plotted against spore concentration and the relation between Tx and protease activity was linear. The viscosity varied and volume percent of particles increased in Tween-80 amended wastewater sludge and particle size (D₅₀) decreased at the end of fermentation. Thus, there was an increase in entomotoxicity at higher suspended solids (30 g/L) as Tween addition improved rheology (viscosity, particle size, surface tension); enhanced maximum growth rate and OUR.     

Direct gene transfer to plant protoplasts by mild sonication

Summary A novel procedure employing mild sonication for transformation of plant protoplasts is described. Transient expression of a chloramphenicol acetyltransferase (CAT) gene in protoplasts of sugar beet (Beta vulgaris L.) and tobacco (Nicotiana tabacum L.) was obtained by a brief exposure of the protoplasts to 20 kHz ultrasound in the presence of plasmid DNA. Maximum levels of CAT activity were achieved by sonication for 500–900 ms at 30–70 W electric power (0.65–1.6 W/cm2 acoustic power). This reduced the viability to 15–20 % and 60 % for sugar beet and tobacco protoplasts, respectively. Up to 12 % (sugar beet) and 81 % (tobacco) of maximum transient expression could be achieved with no significant loss of viability. Protoplasts surviving exposure to ultrasound were found to have a similar long-term viability and to regenerate to micro-calli as untreated protoplasts. Plasmid DNA concentrations of 80–110 g/ml and sucrose concentrations of 21–28 % in the sonication medium were found to be optimal for transient expression.Abbreviations CAT chloramphenicol acetyltransferase     

Effect of low power ultrasonic radiation on anaerobic biodegradability of sewage sludge

The effect of low power ultrasonic radiation on anaerobic biodegradability of sewage sludge was investigated. For this purpose, soluble substances and variation of microbial system of sewage sludge subjected to low power ultrasonic radiation were tested. The well known hydromechanical shear forces and heating effect of low frequency ultrasound plays a major role in the sludge pre treatment process. More, the increase of soluble substance may partly result from the destruction of microbial cell by excess ultrasonic pretreatment, which will inhibit the anaerobic process. By orthogonal tests, the optimal parameters were found to be an exposure time of 15 min, ultrasonic intensity of 0.35 W/cm² and ultrasonic power density of 0.25 W/ml. Under the optimal condition, anaerobic biodegradability of sewage sludge (R_(vss/ss) %) was increased by 67.6%. Consequently, it can be concluded that low power ultrasonic pretreatment is a valid method for improving anaerobic biodegradability of sewage sludge.     

INT-dehydrogenase test for activated sludge process control

Dehydrogenase activity assay of activated sludge using the redox dye 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) was investigated. INT-dehydrogenase activity (INT-DHA) was directly proportional to INT dosage and inversely proportional to bio-mass concentration over limited ranges. INT dosages exceeding 2.5m/M were toxic to dilute activated sludge suspensions. INT-DHA was greatest near pH 9, whereas the peak oxygen uptake rate (OUR) occurred at pH 8. Both INT-DHA and OUR varied inversely with sludge age, but INT-DHA was the more sensitive of the two parameters to this variable. Consistently good and highly significant correlations between INT-DHA and OUR of chlorine stressed activated sludge were found at sludge ages ranging 2.2-7.0 days.     

Ultrasound pretreatment of cassava chip slurry to enhance sugar release for subsequent ethanol production

The use of ultrasound pretreatment to enhance liquefaction and saccharification of cassava chips was investigated. Cassava chip slurry samples were subjected to sonication for 10-40 s at three power levels of low (2 W/mL), medium (5 W/mL), and high (8 W/mL). The samples were simultaneously exposed to enzymes to convert starch into glucose. The cassava particle size declined nearly 40-fold following ultrasonic pretreatment at high power input. Scanning electron micrographs of both unsonicated (control) and sonicated samples showed disruption of fibrous material in cassava chips but did not affect the granular structure of starch. Reducing sugar release improved in direct proportion to the power input and sonication time. The reducing sugar increase was as much as 180% with respect to the control groups. The slurry samples with enzyme addition during sonication resulted in better reducing sugar release than the samples with enzyme addition after sonication. The heat generated during sonication below starch gelatinization temperature apparently had no effect on the reducing sugar release. The reducing sugar yield and energy efficiency of ultrasound pretreated samples increased with total solids (TS) contents. The highest reducing sugar yield of 22 g/100 g of sample and efficiency of 323% were obtained for cassava slurry with 25% TS at high power. The reducing sugar yield at the completion of reaction (R(infinity)) were over twofold higher compared to the control groups. The integration of ultrasound into a cassava-based ethanol plant may significantly improve the overall ethanol yield.     

Biodegradation potential and bacterial diversity of a petrochemical wastewater treatment plant in Iran

An activated sludge treatment was evaluated for its effectiveness in cleaning up a petrochemical wastewater in Iran. For assessing biodegradation potential of activated sludge, seven characteristics of wastewater (temperature, pH, dissolved oxygen, chemical oxygen demand, concentrations of ethylene dichloride, vinyl chloride, and total hydrocarbons) were monitored during six months. It was shown that dominant pollutants in order of magnitude were normal-alkanes (C(10)-C(21)), aromatics, and polycyclic hydrocarbons. The activated sludge treatment revealed maximum reduction of 89%, 99%, 92%, and 80% in COD, ethylene dichloride, vinyl chloride and total hydrocarbons concentrations, respectively. Preliminary screening of culturable petrochemical-degrading microorganisms of the activated sludge resulted in the collection of 67 bacterial and one mold species. Bacterial strains mainly belonged to Pseudomonas, Flavobacterium, Comamonas, Cytophaga, Acidovorax, Sphingomonas, Bacillus and Acinetobacter genera. The isolated mold was identified as Trichoderma sp.     

Study of Biological Activity and Process Stability in Submerged Membrane Bioreactors

Synthetic wastewater was treated in a bench scale submerged membrane bioreactor (SMBR). A long‐term experiment was conducted by varying the sludge residence time (SRT) (10–500 d) and BOD loading (1.3–0.25 kg/m³·d). The biological activity was observed in terms of the oxygen utilization rate (OUR) and adenosine triphosphate (ATP) profile; the process stability was analyzed based on the extent of organic degradation and suction pressure. The microbial population in the SMBR was dependent on the SRT and BOD loading, and its biological activity was increased with an increase in the SRT or BOD loading. At a low feed to microorganism (F/M) ratio (0.06 kg BOD/kg MLSS·d), the sludge production of the reactor was reduced to 0.04 kg MLSS/kg BOD, which is much less than in the conventional activated sludge process (0.4–0.6 kg MLSS/kg BOD). The F/M ratio influenced the biological activity (via ATP and the OUR) significantly at a short SRT (≤90 d). However, the effect of the F/M ratio ceased at a low F/M ratio (≤ 0.07 kg BOD/kg MLSS·d). The accumulation of organics in the SMBR was accompanied with an increase in the supernatant TOC, which caused a high suction pressure and an abrupt change in the operating conditions to process instability. However, the process stability of the SMBR increased with an increase in the SRT and a decrease in the BOD loading along with a concomitant decrease in the biological activity and sludge production.     

Energy-efficient sludge sonication: power and sludge characteristics

This paper improves the energy efficiency for sludge treatment using ultrasound. The sludge organic content, solid concentration, ultrasonic intensity, and time were optimized. A new index, kWh/kgSCOD-increase, was used to evaluate the energy efficiency that covers both sludge characteristics and lysis effectiveness. The results showed that the energy efficiency of sludge sonication was in the range of 102-347kWh/kgSCOD-increase. Higher VS/TS ratio gave higher energy efficiency. Optimal solid concentration (TS) was found to be 20g/L; low TS (5.0g/L) consumed 137% more energy; while higher TS might assimilate the sound energy and decrease the efficiency. The cavitation threshold for sludge was 20-30W/cm(2), below which no sludge lysis was observed. The lowest effective sound intensity (31.4W/cm(2)) was best and saved 20-35% energy compared to higher intensities. The initial first minutes of sonication were most effective; hence intensive-short treatment was preferred.     

Predicting the effects of chlorine on the micro-organisms of filamentous bulking activated sludges

Rapid and definite assessment of the effect that a specific biocide has on a specific case of filamentous bulking sludge is a much-needed tool in activated sludge wastewater treatment. The Live/Dead stain (LIVE/DEAD BacLight) distinguishing "living" and "non-living" cells, a nitrifying activity (NA) test and the oxygen uptake rate (OUR) measurement were examined for their appropriateness to predict the effects of chlorine on filamentous bulking sludges. The study showed the live/dead stain to be relevant for revealing the specific effect of chlorine on the filamentous bacteria of a bulking sludge. However, using live/dead stain alone for the determination of the appropriate chlorine dose against bulking may lead to an underestimation of the damage caused by chlorine to the useful microorganisms in the flocs. Indeed, using the live/dead stain, it was not easy to distinguish dead cells caused by chlorination from those originally present in the flocs The NA test was the most sensitive in detecting damage caused by chlorine to the floc-forming microorganisms. Therefore, for a safer determination of the chlorine dose effective against bulking and protective of the microbial activity of the sludge, the results of this study suggest coupling of the live/dead stain with the NA test and/or the OUR test.     

Effect of a static magnetic field on formaldehyde biodegradation in wastewater by activated sludge

The aim of this study was to determine the impact of a static magnetic field (MF) of 7 mT on formaldehyde (FA) biodegradation by activated sludge in synthetic wastewater. The MF had a positive effect on activated sludge biomass growth and dehydrogenase activity. The influence of the MF on the degradation process was observed with a FA concentration of 2400-2880 mg/l. Decreases in FA concentration and chemical oxygen demand (COD) were greater, by 30% and 26% respectively, than those in the control sample. At initial FA concentrations in raw wastewater of 2400 and 2880 mg/l, a decrease in the wastewater biodegradation efficiency was observed. This resulted in an increase of the ecotoxicity of the effluent to Daphnia magna. The value of the sludge biotic index (SBI) was dependent on the FA concentration in raw wastewater and the induction of the MF.     

Use of Ultrasounds to Reduce the Count of Campylobacter coli in Water

The present study aimed to evaluate the effectiveness of low-frequency ultrasounds applied to eliminate Campylobacter spp. from water. The strains used in this research were isolated from water contaminated with sewage. Campylobacter coli alone was detected in the samples and used for further research. The reference strain C. coli ATCC 33559 was simultaneously tested. The isolate was exposed to ultrasounds at frequencies of 37 kHz and 80 kHz in a continuous operation device with ultrapure deionized water. After 5 min of sonication, the count of C. coli decreased by 5.78% (37 kHz) and 6.27% (80 kHz), whereas the temperature increased by 3°C (37 kHz), and 6°C (80 kHz). After 30 min of sonication, the death rates of bacterial cells were 40.15% (37 kHz) and 55.10% (80 kHz), whereas the temperature reached the maximum values of 36°C (37 kHz), and 39°C (80 kHz). Sonication at the frequency of 80 kHz reduced the bacterial count from 6.86 log CFU/ml to 3.08 log CFU/ml, whereas the frequency of 37 kHz reduced the bacterial count from 6.75 log CFU/ml to 4.04 log CFU/ml. Despite significant differences (p < 0.05) in the number of C. coli cells, the cell death rate remained at the same level. Open in a separate window     

Enhanced Methane Production from Anaerobic Digestion of Disintegrated and Deproteinized Excess Sludge

To improve biogas yield and methane content in anaerobic digestion of excess sludge from the wastewater treatment plant, the sludge was disintegrated by using various methods (sonication, alkaline and thermal treatments). Since disintegrated sludge contains a high concentration of soluble proteins, the resulting metabolite, ammonia, may inhibit methane generation. Therefore, the effects of protein removal from disintegrated sludge on methane production were also studied. As a result, an obvious enhancement of biogas generation was observed by digesting disintegrated sludge (biogas yield increased from 15 to 36 ml/g COD_added·day for the raw excess sludge and the sonicated sludge, respectively). The quality of biogas was also improved by removing proteins from the disintegrated sludge. About 50% (w/w) of soluble proteins were removed from the suspension of disintegrated sludge by salting out using 35 g MgCl₂·6H₂O/l and also by isoelectric point precipitation at pH 3.3. For deproteinized sludge, methane production increased by 19%, and its yield increased from 145 ml/g COD_removed to 325 ml/g COD_removed. Therefore, the yield and quality of biogas produced from digestion of excess sludge can be enhanced by disintegrating the sludge and subsequent protein removal. Revisions requested 14 November 2005; Revisions received 13 January 2006     

Integration of a microbial fuel cell with activated sludge process for energy-saving wastewater treatment: taking a sequencing batch reactor as an example

In the research and application of microbial fuel cell (MFC), how to incorporate MFCs into current wastewater infrastructure is an importance issue. Here, we report a novel strategy of integrating an MFC into a sequencing batch reactor (SBR) to test the energy production and the chemical oxygen demand (COD) removal. The membrane-less biocathode MFC is integrated with the SBR to recover energy from the aeration in the form of electricity and thus reduce the SBR operation costs. In a lab-scale integrated SBR-MFC system, the maximum power production of the MFC was 2.34 W/m(3) for one typical cycle and the current density reached up to 14 A/m(3) . As a result, the MFC contributed to the 18.7% COD consumption of the integrated system and also recovered energy from the aeration tank with a volume fraction of only 12% of the SBR. Our strategy provides a feasible and effective energy-saving and -recovering solution to upgrade the existing activated sludge processes.     

The influence of the energy absorbed from microwave pretreatment on biogas production from secondary wastewater sludge

In this study, microwave treatment is analyzed as a way to accelerate the hydrolysis in anaerobic digestion of municipal wastewater sludge. The influence of the absorbed energy, power and athermal microwave effect on organic matter solubilization and biogas production has been studied. In addition, a novel method that considers the absorbed energy in the microwave system is proposed, in order to obtain comparable experimental results. The absorbed energy is calculated from an energy balance.The highest solubilization was achieved using 0.54 kJ/ml at 1000 W, where an increment of 7.1% was observed in methane production, compared to the untreated sample. Using a higher energy value (0.83 kJ/ml), methane production further increased (to 15.4%), but solubilization decreased. No power influence was found when 0.54 kJ/ml was applied at 1000, 600 and 440 W. Microwave heating was compared to conventional heating in two different experimental setups, providing similar methane yields in all cases.     

O uptake rate measurements as a novel tool to study shear effects on suspended strawberry cells

Despite the same mean volumetric power input of 19 and 47 W/m 3 , the specific oxygen uptake rate (OUR S ) of strawberry ( Fragaria ananassa) cell suspensions was higher in bioreactors equipped with a Rushton Turbine (4.4 and 6.2 3 10 -5 mol O 2 /kg.s) than with an anchor stirrer (2.5 and 4.6 3 10 -5 mol O 2kg.s). The increase in OUR S was caused by stress-activated respiration and appeared to be correlated with the locally dissipated power input. OUR S was corrected for the increase in surface through aggregate break-up and reached a maximum of 6.0 3 10 -5 mol O 2kg.s when agitating with approximately 200 kW/m 3 locally dissipated power input.     

Reducing sludge production and the domination of Comamonadaceae by reducing the oxygen supply in the wastewater treatment procedure of a food-processing factory

Sludge production was reduced remarkably by reducing the dissolved oxygen supply to less than 1 mg/l in the conventional wastewater treatment procedure of a food-processing factory that produced 180 m(3) of wastewater of biochemical oxygen demand (BOD) of about 1,000 mg/l daily. DNA was extracted from the sludge and subjected to PCR amplification. The PCR product was cloned into a plasmid and sequenced. Estimation of the resident bacterial distribution by 16S rDNA sequences before and after improvement of the system suggested a remarkable gradual change in the major bacterial population from Anaerolinaeceae (15.6%) to Comamonadaceae (52.3%), members of denitrifying bacteria of Proteobacteria. Although we did not directly confirm the ability of denitrification of the resulting sludge, a change in the major final electron acceptors from oxygen to nitrate might explain the reduction in sludge production in a conventional activated sludge process when the oxygen supply was limitted.     

The use of ultrasound and gamma-irradiation as pre-treatments for the anaerobic digestion of waste activated sludge at mesophilic and thermophilic temperatures

The effect of ultrasound and gamma-irradiation used as pre-treatments for the anaerobic digestion of waste activated sludge at both mesophilic and thermophilic temperatures was examined. Untreated activated sludge was also subjected to anaerobic digestion at these temperatures as a control. The sonication time was 90 s using a Soniprep 150 (MSE Scientific Instruments) which operated at 23 kHz and had been adjusted to give an output of 47 W and the gamma-irradiation dose was 500 krad. The digesters were operated in a semi-continuous mode, being fed with fresh sludge every 24 h at hydraulic retention times (HRT) of 8, 10 and 12 days. Over the 24 h period the differences between the digesters, in terms of volatile solids (VS) reductions and biogas production, were not statistically significant for any particular set of conditions. Thermophilic digestion performed better than mesophilic digestion in terms of biogas production, VS reductions (except at HRT of 8 days) and specific methane yields and the optimum retention time was 10 days, at both temperatures. When gas production over the initial eight hours (probably the hydrolytic stage) was examined, it was found that the gas production rates for pre-treated sludges were higher than those for untreated sludges. This was most pronounced at thermophilic temperatures and a HRT of 10 days. Sonication did not affect the numbers of faecal coliforms in the sludge. However, gamma-radiation caused a 3-log reduction and, when coupled with mesophilic digestion, gave a product which contained < 100 g(-1) TS. Thermophilic anaerobic digestion produced sludges which contained < 1 g(-1) TS irrespective of any pre-treatment.     

Vitamin addition: an option for sustainable activated sludge process effluent quality

The process performance and metabolic rates of samples of activated sludge dosed with vitamin supplements have been compared. After initial screening, four vitamins and two metals as single supplements and in pairs, were dosed continuously into the mixed liquor of an activated sludge simulation. Toxicity, oxygen demand removal, respiration rates and suspended solids were measured to monitor the effect on process efficiency. It was confirmed experimentally that an industrial wastewater stream did not contain a sufficient supply of micronutrients for efficient biological treatment. This was concluded from the observation that control sludge batches (receiving no supplements) averaged chemical oxygen demand removal efficiency of 58%. Dosing micronutrients into the mixed liquor produced removal efficiencies of up to 69%. Some of the supplements increased the respiration rate of the sludge while some decreased it, indicating a range of stimulatory and inhibitory effects. Complex interactions between micronutrients that were dosed simultaneously were evident. Several positive effects led to the conclusion that micronutrients have the potential to optimise process performance of activated sludge plants treating industrial wastewater. The addition of phosphorus/niacin and molybdenum/lactoflavin removed wastewater components that were toxic to nitrifiers as indicated through toxicity testing, thus protecting downstream nitrification/denitrification treatment processes. Journal of Industrial Microbiology & Biotechnology (2000) 24, 267–274. Received 24 August 1999/ Accepted in revised form 06 January 2000     

Batch biological treatment of nitrogen deficient synthetic wastewater using Azotobacter supplemented activated sludge

Biological treatment of nitrogen deficient wastewaters are usually accomplished by external addition of nitrogen sources to the wastewater which is an extra cost item. As an alternative for effective biological treatment of nitrogen deficient wastewaters, the nitrogen fixing bacterium, Azotobacter vinelandii, was used in activated sludge and also in pure culture. Total organic carbon (TOC) removal performances of Azotobacter-added and free activated sludge cultures were compared at different initial TN/TOC ratios. The rate and extent of TOC removal were comparable for all cultures when initial TN/TOC ratio was larger than 0.12; however, both the rate and extent of TOC removal from nitrogen deficient (TN/TOC<12%) synthetic wastewater were improved by using Azotobacter-added activated sludge as compared to the Azotobacter-free activated sludge culture. More than 90% TOC removal was obtained with pure Azotobacter or Azotobacter-added activated sludge culture from a nitrogen deficient synthetic wastewater.