Evaluation of parameters for monitoring an anaerobic co-digestion process

The system investigated in this study is an anaerobic digester at a municipal wastewater treatment plant operating on sludge from the wastewater treatment, co-digested with carbohydrate-rich food-processing waste. The digester is run below maximum capacity to prevent overload. Process monitoring at present is not extensive, even for the measurement of on-line gas production rate and off-line pH. Much could be gained if a better program for monitoring and control was developed, so that the full capacity of the system could be utilised without the risk of overload. The only limit presently set for correct process operation is that the pH should be above 6.8. In the present investigation, the pH was compared with alkalinity, gas production rate, gas composition and the concentration of volatile fatty acids (VFA). Changes in organic load were monitored in the full-scale anaerobic digester and in laboratory-scale models of the plant. Gas-phase parameters showed a slow response to changes in load. The VFA concentrations were superior for indicating overload of the microbial system, but alkalinity and pH also proved to be good monitoring parameters. The possibility of using pH as a process indicator is, however, strongly dependent on the buffering capacity. In this study, a minor change in the amount of carbohydrates in the substrate had drastic effects on the buffering effect of the system. Received: 21 January 2000 / Received revision: 10 July 2000 / Accepted: 16 July 2000     

Characterization of a prototype industrial on-line analyzer for bicarbonate/carbonate monitoring

In many biological reactors bicarbonate is the major species determining pH buffering capacity, or alkalinity. In anaerobic digesters bicarbonate levels should be within 10 to 50 mM for stable operation. Bicarbonate alkalinity in wastewater treatment processes in routinely measured off-line titrimetrically. Recently we have described the principle of a novel on-line method of measuring bicarbonate alkalinity. In the prototype device described here, a continuous stream (15 cm(3) min(-1)) of the substrate to be monitored was saturated with gaseous CO2, acidified by the addition of excess acid, and the rate of carbon dioxide evolution, proportional to the concentration of bicarbonate/carbonate in the liquid flow, continuously measured by a sensitive gas meter. The instrument was robust and its response was satisfactory for wastewater treatment process control applications, with linearity in the range 5 to 50 mM HCO3(-), a response time in the order of 30 min, and accuracy of the order of 7% in the concentration range 5 to 50 mM sodium bicarbonate. The device was not affected by interference from volatile fatty acids, does not make use of pH probes which in many wastes are subject to fouling, and may form the basis of a digester control strategy. (c) 1994 John Wiley & Sons, Inc.     

Performance evaluation of a mesophilic anaerobic fluidized-bed reactor treating wastewater derived from the production of proteins from extracted sunflower flour

A study of the anaerobic digestion of wastewater derived from the production of protein isolates from extracted sunflower flour was carried out in a laboratory-scale, mesophilic (35 degrees C) fluidized-bed reactor with saponite as bacterial support. Chemical oxygen demand (COD) removal efficiencies in the range of 98.3-80.0% were achieved in the reactor at organic loading rates (OLR) of between 0.6 and 9.3 g COD/I d, hydraulic retention times (HRT) of between 20.0 and 1.1 d and average feed COD concentration of 10.6 g/l. Eighty percent of feed COD could be removed up to OLR of 9.3 g COD/l d. The yield coefficient of methane production was 0.33 l of methane (at STP) per gram of COD removed and was virtually independent of the OLR applied. Because the buffering capacity of the experimental system was maintained at favorable levels with excess total alkalinity present at all loadings, the rate of methanogenesis was not affected by loading. The experimental data indicated that a total alkalinity in the range of 2,000-2,460 mg/l as CaCO3 was sufficient to prevent the pH from dropping to below 7.0 for OLR of up to 9.3 g COD/l d. The volatile fatty acid (VFA) levels and the VFA/alkalinity ratio were lower than the suggested limits for digester failure (0.3-0.4) for OLR and HRT up to 9.3 g COD/l d and 1.1 d, respectively. For a HRT of 0.87 d (OLR of 12.1 g COD/l d) the start of acidification was observed in the reactor.     

pH对鲤鳃部微环境铜形态的影响

利用Playle等设计的装置研究了不同pH条件及不同的铜暴露水平下鲤鳃部微环境pH值和粘液分泌量的变化,在此基础上用化学平衡计算方法探讨鱼鳃微环境中铜形态的变化。结果表明,在特定的pH和铜浓度范围内,鱼鳃微环境平衡pH略低于7,鱼鳃粘液分泌量随铜暴露水平提高而增加,与此同时,虽然CO2分泌量有所增加,没有观察到微环境pH随铜暴露量的变化。pH的升高,导致水相优势的游离态铜让位给羟基络合铜,粘液络合态铜的分额逐渭降低,但不会改变其络合当量与条件稳定络合常数。由于鱼鳃微环境对pH改变的调节作用,铜的形态分布变化幅度低于外部环境。     

Anaerobic treatment of Tequila vinasses in a CSTR-type digester

Tequila industries in general produce great volumes of effluents with high pollutant loads, which are discharged (untreated or partially treated) into natural receivers, thus causing severe environmental problems. In this contribution, we propose an integrated system as a first step to comply with the Mexican ecological norms and stabilize the anaerobic treatment of Tequila vinasses with main design criteria: simple and easy operation, reduce operating time and associated costs (maintenance), integrated and compact design, minimal cost of set-up, start-up, monitoring and control. This system is composed of a fully instrumented and automated lab-scale CSTR-type digester, on-line measuring devices of key variables (pH, temperature, flow rates, etc.), which are used along with off-line readings of chemical oxygen demand (COD), biogas composition, alkalinity and volatile fatty acids to guarantee the operational stability of the anaerobic digestion process. The system performance was evaluated for 200 days and the experimental results show that even under the influence of load disturbances, it is possible to reduce the COD concentration to 85% in the start-up phase and up to 95% during the normal operation phase while producing a biogas with a methane composition greater than 65%. It is also shown that in order to maintain an efficient treatment, the buffering capacity (given by the alkalinity ratio, α = intermediate alkalinity/total alkalinity) must be closely monitored.     

Mesophilic anaerobic digestion in a fluidised-bed reactor of wastewater from the production of protein isolates from chickpea flour

A study of the anaerobic digestion of wastewater derived from the production of protein isolates from chickpea flour was carried out in a laboratory-scale, mesophilic (35 °C) fluidised-bed reactor with saponite as bacterial support. Soluble chemical oxygen demand (SCOD) removal efficiencies in the range of 96.8–85.2% were achieved in the reactor at organic loading rates (OLR) of between 0.58 and 2.10 g chemical oxygen demand (COD)/l per day, hydraulic retention times (HRT) of between 14.9 and 4.5 days and average feed COD concentration of 9.1 g/l. Eighty-five percent of feed COD could be removed up to OLR of 2.1 g COD/l per day. The yield coefficient of methane production was 0.34 l of methane (at STP) per gram COD removed and was virtually independent of the OLR applied. Because the buffering capacity of the experimental system was maintained at favourable levels with excess total alkalinity present at all loadings, the rate of methanogenesis was not affected by loading. Experimental data indicated that a total alkalinity in the range of 1090–2130 mg/l as CaCO₃ was sufficient to prevent the pH from decreasing to below 7.2 for OLR of up to 2.7 g COD/l per day. The volatile fatty acid (VFA) levels and the VFA/alkalinity ratio were lower than the suggested limits for digester failure (0.3–0.4) for OLR and HRT up to 2.7 g COD/l per day and 3.5 days, respectively. For a HRT of 2.8 days (OLR of 3.00 g COD/l per day) the start of acidification was observed in the reactor.     

Advanced monitoring and supervision of biological treatment of complex dairy effluents in a full-scale plant

The operation of a wastewater treatment plant treating effluents from a dairy laboratory was monitored by an advanced system. This plant comprises a 12 m(3) anaerobic filter (AF) reactor and a 28 m(3) sequential batch reactor (SBR) coupled in series and is equipped with the following on-line measurement devices: biogas flow meter, feed and recycling flow meters, temperature sensor, dissolved oxygen analyzer, and redox meter. Other parameters such as chemical oxygen demand (COD), volatile fatty acids (VFA), etc. were determined off-line. The plant has been in operation for 634 days, the influent flow rate being 6-8 m(3)/d. COD concentration of the influent ranged between 8 and 12 kg COD/m(3), resulting in COD values in the effluent around 50-200 mg/L. The behavior of the system was studied using the set of measurements collected by the data acquisition program especially developed for this purpose. Monitoring of variables such as anaerobic reactor temperature permitted the detection and prevention of several failures such as temperature shocks in the AF reactor. Besides, off-line measurements such as the alkalinity or the VFA content, together with the on-line measurements, provided immediate information about the state of the plant and the detection of several anomalies, such as organic overloads in the SBR, allowing the implementation of several fast control actions.     

Influence of pH and temperature on soluble substrate generation with primary sludge fermentation

The study was undertaken to evaluate the effect of pH and temperature control on the generation of soluble fermentation products from primary sludge. The effect was tested by running parallel experiments under pH and temperature controlled and uncontrolled conditions. In fermentation experiments conducted at 20 degrees C without pH control, the average soluble COD release was 14 mg per liter of wastewater treated, representing a potential increase of 5% in the biodegradable COD content of the primary sedimentation effluent. The corresponding average VFA generation was 9.2mg COD l(-1). The nutrient release was practically negligible and stayed at 0.4 mg l(-1) for nitrogen and 0.1mg l(-1) for phosphorus. Acetic acid accounted more than 45% of the generated VFA in all experimental runs. The acetic acid content of the VFA decreased with increasing initial VSS concentrations and higher pH levels. VFA generation by fermentation was significantly affected with temperature and pH control. Temperature change between 10 and 24 degrees C induced a five-fold increase in VFA generation, from 610 mg l(-1) at 10 degrees C to 2950 mg l(-1) at 24 degrees C.     

On-line near infrared monitoring of glycerol-boosted anaerobic digestion processes: evaluation of process analytical technologies

A study of NIR as a tool for process monitoring of thermophilic anaerobic digestion boosted by glycerol has been carried out, aiming at developing simple and robust Process Analytical Technology modalities for on-line surveillance in full scale biogas plants. Three 5 L laboratory fermenters equipped with on-line NIR sensor and special sampling stations were used as a basis for chemometric multivariate calibration. NIR characterisation using Transflexive Embedded Near Infra-Red Sensor (TENIRS) equipment integrated into an external recurrent loop on the fermentation reactors, allows for representative sampling, of the highly heterogeneous fermentation bio slurries. Glycerol is an important by-product from the increasing European bio-diesel production. Glycerol addition can boost biogas yields, if not exceeding a limiting 5-7 g L(-1) concentration inside the fermenter-further increase can cause strong imbalance in the anaerobic digestion process. A secondary objective was to evaluate the effect of addition of glycerol, in a spiking experiment which introduced increasing organic overloading as monitored by volatile fatty acids (VFA) levels. High correlation between on-line NIR determinations of glycerol and VFA contents has been documented. Chemometric regression models (PLS) between glycerol and NIR spectra needed no outlier removals and only one PLS-component was required. Test set validation resulted in excellent measures of prediction performance, precision: r(2) = 0.96 and accuracy = 1.04, slope of predicted versus reference fitting. Similar prediction statistics for acetic acid, iso-butanoic acid and total VFA proves that process NIR spectroscopy is able to quantify all pertinent levels of both volatile fatty acids and glycerol.     

On-line monitoring of IPTG induction for recombinant protein production using an automatic pH control signal

The response of IPTG induction was investigated through the monitoring of the alkali consumption rate and buffer capacity during the cultivation of recombinantE. coli BL21(DE3) harboring the plasmid pRSET-LacZ under the control oflac promoter. The rate of alkali consumption increased along with cell growth, but declined suddenly after approximately 0.2 h of IPTG induction. The buffer capacity also declined after 0.9 h of IPTG induction. The profile of buffer capacity seems to correlate with the level of acetate production. The IPTG response was monitored only when introduced into the mid-exponential phase of bacterial cell growth. The minimum concentration of IPTG for induction, which was found out to be 0.1 mM, can also be monitored on-line andin-situ. Therefore, the on-line monitoring of alkali consumption rate and buffer capacity can be an indicator of the metabolic shift initiated by IPTG supplement, as well as for the physiological state of cell growth.     

Dual excitation ratiometric fluorescent pH sensor for noninvasive bioprocess monitoring: development and application

The development and application of a fluorescent excitation-ratiometric, noninvasive pH sensor for continuous on-line fermentation monitoring is presented. The ratiometric approach is robust and insensitive to factors such as source intensity, photobleaching, or orientation of the patch, and since measurements can be made with external instrumentation and without direct contact with the patch, detection is completely noninvasive. The fluorescent dye 8-hydroxy-1,3,6-pyrene trisulfonic acid was immobilized onto Dowex strongly basic anion-exchange resin, which was subsequently entrapped into a proton-permeable hydrogel layer. The sensor layer was polymerized directly onto a white microfiltration membrane backing that provided an optical barrier to the fluorescence and scatter of the fermentation medium. The ratio of emission intensity at 515 nm excited at 468 nm to that excited at 408 nm correlated well with the pH of clear buffers, over the pH range of 6-9. The sensor responded rapidly (<9 min) and reversibly to changes in the solution pH with high precision. The sterilizable HPTS sensor was used for on-line pH monitoring of an E. coli fermentation. The output from the indwelling sensor patch was always in good agreement with the pH recorded off-line with an ISFET probe, with a maximum discrepancy of 0.05 pH units. The sensor is easily adaptable to closed-loop feedback control systems.     

Use of limestone for pH control in autotrophic denitrification: continuous flow experiments in pilot-scale packed bed reactors

The sulfur-utilizing autotrophic denitrification process consumes about 4 g alkalinity (as CaCO(3)) per g NO(3)-N reduced resulting in a decrease of pH. Using limestone as an alkalinity source to control the pH, autotrophic denitrification of synthetic wastewater with varying alkalinity to NO(3)-N ratios was evaluated in pilot-scale packed bed reactors operating in the upflow mode, which contained limestone and sulfur granules in different volumetric ratios. The results demonstrated that limestone supplies effective buffering capacity, if the initial alkalinity of the wastewater is insufficient for complete denitrification. The alkalinity supplied by limestone is a function of hydraulic retention time and the pH, which in turn depends on the extent of biological denitrification and the initial alkalinity to NO(3)-N ratio in the wastewater. The dissolution rate of limestone is inversely proportional to pH for pH values lower than 7.1. It was found that the ratio of influent alkalinity to theoretically required alkalinity in the wastewater should not be lower than 0.5 in order to prevent a decrease in nitrate removal performance. Based on the established chemical-biological interactive relationships, a multilayer approach was proposed to determine the optimum sulfur:limestone ratio for nitrate removal under steady state conditions, taking into account the characteristics of the influent wastewater.     

Volatile fatty acids as indicators of process imbalance in anaerobic digestors

In continuously stirred tank reactor experiments, with manure as substrate at thermophilic temperatures, the use of volatile fatty acids (VFA) as process indicators was investigated. Changes in VFA level were shown to be a good parameter for indicating process instability. The VFA were evaluated according to their relative changes caused by changes in hydraulic loading, organic loading or temperature. Butyrate and isobutyrate together were found to be particularly good indicators. Butyrate and isobutyrate concentrations increased significantly 1 or 2 days after the imposed perturbation, which makes these acids suitable for process monitoring and important for process control of the anaerobic biological system. In addition it was shown in a batch experiment that VFA at concentrations up to 50 mM did not reduce the overall methane production rate. This showed that VFA accumulation in anaerobic reactors was the result of process imbalance, not the cause of inhibition, thus justifying the use of VFA as process indicators.     

Criteria for evaluating calcium carbonate from the point of view of chlortetracycline biosynthesis]

Calcium carbonate is added to fermentation media in biosynthesis of tetracyclines for providing definite pH values and binding tetracycline into insoluble complexes. Seven different samples were studied with respect to their physical properties, such as the microscopic size of the particles, their form, capacity for agglomeration, specific volume, rate of the particle precipitation and chemical properties, such as purity, buffer capacity, effect on the medium pH before and after sterilization. The above properties were studied in comparison with activity chlortetracycline biosynthesis. Microfine calcium carbonate proved to be the best from the point of view of productivity of Str. aureofaciens. With its use the activity of the culture fluid increased by 20 per cent as compared to the other samples. The titration curve of the sample had the lowest bend.     

Customized design of electronic noses placed on top of air-lift bioreactors for in situ monitoring the off-gas patterns

A specially designed electronic nose was coupled to an air-lift bioreactor in order to perform on-line monitoring of released vapors. The sensor array was placed at the top of the bioreactor sensing the headspace in equilibrium with the evolving liquor at any time without the need of aspiration and pumping of gases into a separated sensor chamber. The device was applied to follow the off-gas of a bioreactor with Acidithiobacillus thiooxidans grown on beds of elemental sulfur under aerobic conditions. Evolution was monitored by acid titration, pH and optical density measurements. The electronic nose was capable to differentiate each day of reactor evolution since inoculation within periods marked off culture medium replacements using multivariate data analysis. Excellent discrimination was obtained indicating the potentiality for on-line monitoring in non-perturbed bioreactors. The prospects for electronic nose/bioreactor merging are valuable for whatever the bacterial strain or consortium used in terms of scent markers to monitor biochemical processes.     

Nuclear magnetic resonance titration curves of histidine ring protons. Ribonuclease S-peptide and S-proteins.

The histidine C-2 proton NMR titration curves of ribonuclease S-peptide (residues 1 to 20) and S-protein (residues 21 to 124) are reported. Although S-protein contains 3 histidine residues, four discrete resonances are observed to titrate. One of these arises from the equivalent histidine residues of unfolded S-protein. The variation in area of the four resonances indicate that there is a reversible pH-dependent equilibrium between the folded and unfolded forms of S-protein, with some unfolded material being present at most pH values. Two of the resonances of the folded S-protein can be assigned to 2 of the histidine residues, 48 and 105, from the close similarity of their titration curves to those in ribonuclease. These similarities indicate a homology of portions of the folded conformation of S-protein to that of ribonuclease in solution. These results indicate that the complete amino acid sequence is not required to produce a folded conformation similar to the native globular protein, and they appear to eliminate the possibility that proteins fold from their NH2 terminus during protein synthesis. The low pH inflection present in the titration curve assigned to histidine residue 48 in ribonuclease is absent from this curve in S-protein. This is consistent with our previous conclusion that this inflection arises from the interaction of histidine 48 with aspartic acid residue 14, which is also absent in S-protein. The third titrating resonance of native S-protein is assigned to the remaining histidine residue at position 119. The properties of this resonance are not identical with either of the titration curves of the active site histidine residues 12 and 119 of ribonuclease. The resonance assigned to histidine 119 is the only one significantly affected on the addition of sodium phosphate to S-protein, indicating that some degree of phosphate binding occurs. In both the absence and presence of phosphate this curve also lacks the low pH inflection observed in the histidine 119 NMR titration curve in ribonuclease. This difference presumably arise from a conformational between ribonuclease and the folded S-protein involving a carboxyl group.     

Inactivation of Salmonella typhi by high levels of volatile fatty acids during anaerobic digestion

Survival of Salmonella typhi was investigated in an anaerobic digester for cattle dung with volatile fatty acid (VFA) levels of 5000 mg l(-1) and pH 6.0. The organism was added to the digester only once in the first experiment and daily in the other. Survival was monitored on alternate days. In the single dose experiment, the counts of Salm. typhi declined rapidly and the pathogen was completely eliminated within 12 d in the experimental digester (VFA ca 5000 mg l(-1) and pH 6.0), whereas 26 d were required in the control digester (VFA ca 100 mg l(-1) and pH 6.8). T90 values for the experimental and control digesters were 2.44 d and 4.80 d, respectively. In the daily dose experiment, a four log reduction in the pathogen count was observed in the experimental digester, but only a two log reduction in the control digester at the end of the experimental period. The mean T90 values for the experimental and the control digester were 4.22 d and 18.63 d, respectively. In both the experiments, statistical analysis of the data showed significant differences in the survival pattern of Salm. typhi in the two digesters.     

Application of response surface methodology for optimization of acidogenesis of cattail by rumen cultures

Acidogenesis of cattail using rumen cultures was carried out to produce volatile fatty acids (VFA) in this study. The influences of pH and substrate concentration on cattail degradation, VFA yield and microbial growth were investigated by using response surface methodology (RSM). Experimental results showed that a low substrate concentration and pH of 6.9 were optimal for acidogenesis of cattail. The highest cattail degradation efficiency, VFA yield and microbial yield were 75.9%, 0.41 g/g VS and 0.110 g/g VS, respectively. Further experiments confirmed that the main VFA in the acidogenesis of cattail were acetate, propionate and butyrate, while i-butyrate, valerate and i-valerate were also produced at low levels. The results suggested that acidogenesis using rumen cultures is a promising method for cattail disposal.